Acknowledgments

The development of the DspikeIn package was made possible through the generous and pioneering efforts of the R and Bioconductor communities. We gratefully acknowledge the developers and maintainers of the following open-source packages, whose tools and infrastructure underpin our work: Core infrastructure & data manipulation: methods, stats, utils, graphics, grDevices, data.table, dplyr, tibble, tidyr, reshape2, matrixStats, rlang, S4Vectors, grid, officer, xml2 Statistical analysis & modeling: DESeq2, edgeR, limma, randomForest, microbiome Phylogenetics & microbiome structure: phyloseq, TreeSummarizedExperiment, SummarizedExperiment, phangorn, ape, DECIPHER, msa, Biostrings Network and graph analysis: igraph, ggraph Visualization & layout design: ggplot2, ggrepel, ggpubr, ggnewscale, ggalluvial, ggtree, ggtreeExtra, ggstar, ggridges, patchwork, scales, RColorBrewer, flextable

These tools collectively empowered us to build a reproducible, modular, and extensible platform for robust absolute abundance quantification in microbial community analysis. We further acknowledge the broader scientific community working on absolute microbial quantification, spike-in calibration, and compositional data analysis, whose foundational insights directly informed the design and conceptual framework of DspikeIn.

if (!requireNamespace("BiocManager", quietly = TRUE))
  install.packages("BiocManager")
BiocManager::install("DspikeIn")

library(DspikeIn)

DspikeIn

The DspikeIn package supports both phyloseq and TreeSummarizedExperiment formats to streamline microbial quantification across diverse experimental setups. It accommodates either a single spike-in taxon or synthetic community taxa with variable or equal spike-in volumes and copy numbers. The package offers a comprehensive suite of tools for absolute abundance (AA) quantification, addressing challenges through ten core functions: 1) validation of spiked species, 2) data preprocessing, 3) system-specific spiked species retrieval, 4) scaling factor calculation, 5) conversion to absolute abundance, 6) bias correction and normalization, 7) performance assessment, and 8) taxa exploration and filtering 9) network topology assessment 10) further analyses and visualization. For more information please check doi.org/10.1093/ismejo/wraf150

DspikeIn requirements

Important: DspikeIn operates on a taxonomy table with exactly seven ranks: Kingdom Phylum Class Order Family Genus Species. colnames(taxmat) <- c(“Kingdom”, “Phylum”, “Class”, “Order”, “Family”, “Genus”, “Species”) Please remove or rename extra ranks (e.g., Strain, Subfamily) before running DspikeIn.

library(SummarizedExperiment)
library(DspikeIn)

# DspikeIn requirements
# --------------------
# DspikeIn operates on a taxonomy table with exactly seven ranks:
# Kingdom  Phylum  Class  Order  Family  Genus  Species

expected <- c("Kingdom", "Phylum", "Class", "Order", "Family", "Genus", "Species")
actual <- colnames(as.data.frame(SummarizedExperiment::rowData(tse)))

if (!identical(expected, actual)) {
  stop(
    paste0(
      "DspikeIn requires exactly 7 taxonomic ranks in this order:\n  ",
      paste(expected, collapse = "  "),
      "\n\nYour taxonomy columns are:\n  ",
      paste(actual, collapse = ", "),
      "\n\nPlease remove or rename extra ranks (e.g., 'Strain', 'Subfamily') ",
      "to match the required format."
    )
  )
}

To build TreeSummarizedExperiment file please refer to

github.com/markrobinsonuzh/TreeSummarizedExperiment

library(phyloseq)
# Get path to external data folder
extdata_path <- system.file("extdata", package = "DspikeIn")
list.files(extdata_path)

## [1] "Complete.graphml" "NoBasid.graphml"  "NoHubs.graphml"   "Ref.fasta"       
## [5] "Sample.fasta"

# and
data("physeq_16SOTU", package = "DspikeIn")
length(taxa_names(physeq_16SOTU))

## [1] 9242

new_ids <- paste0("OTU", seq_along(taxa_names(physeq_16SOTU)))
taxa_names(physeq_16SOTU) <- new_ids

# Verify
head(taxa_names(physeq_16SOTU))

## [1] "OTU1" "OTU2" "OTU3" "OTU4" "OTU5" "OTU6"

# [1] "OTU1" "OTU2" "OTU3" "OTU4" "OTU5" "OTU6"



tse_16SOTU <- convert_phyloseq_to_tse(physeq_16SOTU)
tse_16SOTU <- tidy_phyloseq_tse(tse_16SOTU)

Do all detected sample spike-in sequences cluster with the reference, and are their branch lengths statistically similar, supporting a common ancestor?

spike-in validation

All sample-derived sequences are forming a clade with the reference. We look for a monophyletic grouping of spike-in OTUs The clade is strongly supported (bootstrap around 100 percentage). The branch lengths and distances are in a biologically plausible range.

# Use the Neighbor-Joining method  based on a Jukes-Cantor distance matrix and plot the tree with bootstrap values.
# we compare the Sanger read of Tetragenococcus halophilus with the FASTA sequence of Tetragenococcus halophilus from our phyloseq object.

library(Biostrings)
library(TreeSummarizedExperiment)
library(SummarizedExperiment)
library(DspikeIn)


# Filter TSE object to keep only Bacteria and Archaea
tse_16SOTU <- tse_16SOTU[
  rowData(tse_16SOTU)$Kingdom %in% c("Bacteria", "Archaea"),]


library(Biostrings)
# 
# # Subset the TSE object to include only Tetragenococcus
# tetragenococcus_tse <- tse_16SOTU[
#   rowData(tse_16SOTU)$Genus == "Tetragenococcus" &
#     !is.na(rownames(tse_16SOTU)) &
#     rownames(tse_16SOTU) != "", ]
# 
# ref_sequences <- referenceSeq(tetragenococcus_tse)
# 
# # Convert to DNAStringSet if needed
# ref_sequences <- Biostrings::DNAStringSet(ref_sequences)
# Biostrings::writeXStringSet(ref_sequences, filepath = "Sample.fasta")

ref_fasta <- system.file("extdata", "Ref.fasta", package = "DspikeIn")
sample_fasta <- system.file("extdata", "Sample.fasta", package = "DspikeIn")



result <- validate_spikein_clade(
  reference_fasta = ref_fasta,
  sample_fasta = sample_fasta,
  bootstrap = 200,
  output_prefix = NULL)

## use default substitution matrix

result$tree_plot

Did spike-ins behave as expected across all samples?

Tip labels= OTU/ASV names Branch length numbers= Actual evolutionary distances (small = very similar) Prevalence stars How frequently the OTU occurs across samples Blue bar ring= Log10 mean abundance Outer colored tiles= The metadata variable you choose (e.g., Animal.type)

library(ggstar)
library(ggplot2)

# Filter your object to only include spike-in taxa beforehand:
# We changed the OTU IDs for easy detection
# Big stars = detected in many samples
# Small stars = rarely detected
# log10(Mean Abundance) Bars= Color intensity reflects mean abundance.
# The log-transformed average abundance of each OTU across all samples where it appears.
# Extreme blue may signal unintended over-representation.
# Metadata Ring = factor of your interest e.g. Animal.type
# Each OTU is colored by where it was observed.
# Branch length numbers=    Actual evolutionary distances (small = very similar)


library(DspikeIn)
library(TreeSummarizedExperiment)

# ---- 1. Subset taxa where Genus is Tetragenococcus ----
spikein_tse <- tse_16SOTU[
  rowData(tse_16SOTU)$Genus == "Tetragenococcus", ]


# ---- 2. Diagnostic plot (tree-based) ----
ps <- plot_spikein_tree_diagnostic(
  obj = spikein_tse,
  metadata_var = "Animal.type",
  save_plot = FALSE )

print(ps)

Pre_processing

merges monophyletic ASVs/OTUs. Why We Merge Similar ASVs/OTUs? In amplicon sequencing, multiple Amplicon Sequence Variants (ASVs) or Operational Taxonomic Units (OTUs) often map to the same taxonomic species due to: sequencing noise, PCR amplification artifacts, or minor intra-strain variations that do not represent biologically distinct taxa. When performing absolute quantitation (such as spike-in recovery), these subtle sequence variants should not be treated as independent species, since they originate from the same biological organism. Retaining them as separate entries can artificially fragment read counts and distort abundance estimates. Therefore, this function pre-processes the dataset by merging all ASVs/OTUs that belong to the same declared taxon into a single representative unit, while keeping the phylogenetic tree, taxonomy, and reference sequences consistent. This ensures that the subsequent quantitation step reflects true organism-level abundance, not technical redundancy.

When to use: As the first preprocessing step before calculating spike-in recovery percentages or absolute quantitation metrics.

Spiked species and related parameters should be defined

#                      PREREQUISITE FOR 16S & CALCULATE SPIKED %

# Define spiked species and related parameters**

library(flextable)
library(DspikeIn)
library(TreeSummarizedExperiment)
library(SummarizedExperiment)
library(dplyr)

# Define spike-in parameters
spiked_cells <- 1847
species_name <- spiked_species <- c("Tetragenococcus_halophilus", "Tetragenococcus_sp.")

merged_spiked_species <- "Tetragenococcus"

# If you prefer Genus-level matching
# species_name <- spiked_species <- c("Tetragenococcus")
# merged_spiked_species <- "Tetragenococcus"



# --------------------------------------------
# Subset taxa for the spike-in species
tse_16SOTU_spiked_taxa <- tse_16SOTU[rowData(tse_16SOTU)$Species %in% species_name, ]

# Subset samples based on spike-in volume (e.g., "1" or "2")
tse_16SOTU_spiked_samples <- tse_16SOTU[, colData(tse_16SOTU)$spiked.volume %in% c("2", "1") ]

# --------------------------------------------
# Merge spiked OTUs using the DspikeIn function
# merges monophyletic ASVs/OTUs
# The function Pre_processing_species() merges ASVs/OTUs of an spiked species using "sum" or "max" methods, 
# preserving taxonomic, phylogenetic, and sequencing data.

output_rds <- file.path(tempdir(), "merged_tse_sum.rds")

Spiked_16S_sum_scaled <- Pre_processing_species(
  tse_16SOTU_spiked_samples,
  species_name,
  merge_method = "sum",
  output_file = output_rds)

Calculate the percentage of spiked species retrieval

Why and What It Measures? Spike-in standards provide an internal quantitative reference for microbial load normalization. After merging the spike-in species, we can evaluate how many sequencing reads in each sample correspond to these known spike-in taxa. This value serves as a recovery control, reflecting both: the accuracy of DNA extraction and sequencing, and the efficiency of downstream normalization. By expressing spike-in reads as a percentage of total reads per sample, we can detect samples with technical failure (too low recovery) or overrepresentation (possible contamination or pipetting bias etc.). This metric provides an essential quality-control checkpoint: samples with spike-in recovery outside the expected range (e.g., <0.1% or >20%) may indicate experimental anomalies or uneven DNA extraction efficiency. The resulting table allows quick identification of samples passing or failing QC thresholds, ensuring that downstream absolute quantitation is based on reliable input data.

# * Calculate the percentage of spiked species retrieval per sample*

library(mia)         
library(dplyr)
library(SummarizedExperiment)

# Subset TSE to remove blanks 
Spiked_16S_sum_scaled_filtered <- Spiked_16S_sum_scaled[, colData(Spiked_16S_sum_scaled)$sample.or.blank != "blank"]

# Calculate spike-in retrieval percentage
Perc <- calculate_spike_percentage(
  Spiked_16S_sum_scaled_filtered,
  merged_spiked_species,
 #output_file = "output_tse.docx",
  passed_range = c(0.1, 20)
)

head(Perc)

##              Sample Total_Reads Spiked_Reads Percentage Result
## 1 STP1719.20422_S47       19142        14554 76.0317626 failed
## 2  STP213.20423_S59        8462           83  0.9808556 passed
## 3  STP268.20424_S71        6968           17  0.2439724 passed
## 4  STP544.20419_S11        2340         2259 96.5384615 failed
## 5  STP570.20420_S23        2647          822 31.0540234 failed
## 6  STP579.20421_S35        5193         1759 33.8725207 failed

spiked species retrieval is system-dependent

Spike-in controls provide a way to assess the quantitative reliability of metagenomic or amplicon sequencing workflows. However, the expected recovery percentage of spike-in reads is not universal, it depends on the sequencing platform, extraction chemistry, and the complexity of the native microbial community. As discussed in our paper (doi.org/10.1093/ismejo/wraf150), the acceptable range of spike-in recovery is system-dependent and should be determined empirically from each dataset. This function (regression_plot()) supports that step by visualizing the relationship between observed spike-in abundance and measured total reads, stratified by recovery percentage ranges. The regression_plot() generates a scatter plot of log-transformed variables (typically Observed vs Total_Reads_spiked), overlays a linear regression fit, and facets the data according to custom recovery ranges (e.g., 010%, 1030%, 3050%, etc.).

Interpretation: A strong linear trend (high R, low p-value) within a range indicates consistent quantitative behavior. Deviations or flattening slopes suggest technical bias or over-amplification. The empirically stable range can then be used to adjust the passed_range argument in calculate_spike_percentage() for future analyses. Why this matters: By empirically determining recovery behavior instead of assuming a fixed threshold, users ensure that spike-in QC is tailored to their experimental conditions, improving both accuracy and comparability of absolute quantitation results.

# The acceptable range of spiked species retrieval is system-dependent
# Spiked species become centroid of the community (Distance to Centroid)
# Spiked species become dominant and imbalance the community (Evenness)

# What range of spiked species retrieval is appropriate for your system?
# Calculate Pielou's Evenness using Shannon index and species richness (Observed)
# Hill number q = 1 = exp(Shannon index), representing the effective number of equally abundant species. Hill is weighted by relative abundance, so dominant species have more influence.
# Unlike Pielou's evenness, this metric is not normalized by richness and it shows Effective number of common species

library(TreeSummarizedExperiment)
library(SummarizedExperiment)
library(dplyr)
library(tibble)
library(microbiome)
library(mia)
library(vegan)
library(S4Vectors)
library(ggplot2)


# --- 1. Extract current metadata from TSE ---
metadata <- colData(Spiked_16S_sum_scaled) %>%
  as.data.frame() %>%
  rownames_to_column("Sample")

# --- 2. Add spike-in reads (Perc) ---
# Ensure Perc has 'Sample' column and matching format
metadata <- dplyr::left_join(metadata, Perc, by = "Sample")

# --- 3. Estimate alpha diversity indices and extract ---
Spiked_16S_sum_scaled <- mia::addAlpha(
  Spiked_16S_sum_scaled,
  index = c("observed", "shannon", "pielou", "hill")
)

alpha_df <- colData(Spiked_16S_sum_scaled) %>%
  as.data.frame() %>%
  rownames_to_column("Sample") %>%
  select(Sample, observed, shannon, pielou, hill)

metadata <- dplyr::left_join(metadata, alpha_df, by = "Sample")

# --- 4. Compute Bray-Curtis distance to centroid ---
otu_mat <- assay(Spiked_16S_sum_scaled, "counts")
otu_mat <- t(otu_mat)  # samples as rows
otu_mat_rel <- vegan::decostand(otu_mat, method = "total")

centroid_profile <- colMeans(otu_mat_rel)

dist_to_centroid <- apply(otu_mat_rel, 1, function(x) {
  vegan::vegdist(rbind(x, centroid_profile), method = "bray")[1]
})

# Match and assign distances
metadata$Dist_to_Centroid <- dist_to_centroid[metadata$Sample]

# --- 5. Assign updated metadata to TSE ---
metadata <- column_to_rownames(metadata, var = "Sample")
metadata <- metadata[colnames(Spiked_16S_sum_scaled), , drop = FALSE]  # ensure correct order and size

colData(Spiked_16S_sum_scaled) <- S4Vectors::DataFrame(metadata)


# 4. Regression Plots: Diversity vs. Spike-in Reads
# ============================================================================

# Pielous Evenness
plot_object_pielou <- regression_plot(
  data = metadata,
  x_var = "pielou",
  y_var = "Spiked_Reads",
  custom_range = c(0.1, 20, 30, 40, 50, 60, 100),
  plot_title = "Pielous Evenness vs. Spike-in Reads"
)

# Hill Number (q = 1)
plot_object_hill <- regression_plot(
  data = metadata,
  x_var = "hill",
  y_var = "Spiked_Reads",
  custom_range = c(0.1, 10, 20, 30, 100),
  plot_title = "Hill Number (q = 1) vs. Spike-in Reads"
)

plot_object_hill

# Distance to Centroid
plot_object_centroid <- regression_plot(
  data = metadata,
  x_var = "Dist_to_Centroid",
  y_var = "Spiked_Reads",
  custom_range = c(0.1, 20, 30, 40, 50, 60, 100),
  plot_title = "Distance to Global Centroid vs. Spike-in Reads"
)


# Interpretation
# ============================================================================
# - Pielou's evenness is normalized by richness; useful for detecting imbalance.
# - Hill number q = 1 gives effective number of common species; sensitive to dominance.
# - Distance to centroid in full Bray-Curtis space shows deviation from the average community.

Spiked species and related parameters for ITS

#                      PREREQUISITE FOR ITS & CALCULATE SPIKED %


# Define spiked species and related parameters**


# Define the spiked species
# spiked_cells <- 733
# species_name <- spiked_species <- merged_spiked_species <- "Dekkera_bruxellensis"

# Subset taxa for spiked species
# Dekkera <- phyloseq::subset_taxa(
# physeq_ITSOTU,
# Species %in% species_name)

# hashcodes <- row.names(phyloseq::tax_table(Dekkera))

# Subset samples based on spiked volume
# physeq_ITSOTU_spiked <- phyloseq::subset_samples(physeq_ITSOTU, spiked.volume %in% c("2", "1"))

# if TSE format
# tse_ITSOTU <- convert_phyloseq_to_tse(physeq_ITSOTU)
# physeq_ITSOTU_spiked <- tse_ITSOTU[, tse_ITSOTU$spiked.volume %in% c("2", "1")]

Calculating Scaling Factors

The calculate_spikeIn_factors() function estimates bias-corrected scaling factors for absolute microbiome quantitation using spike-in standards. The ratio between known and observed spike-in counts yields a sample-specific correction factor that normalizes total microbial abundances to absolute scale. This whole-cell spike-in approach corrects for variations in DNA extraction efficiency, amplification yield, and sequencing depth, yielding a unified, biologically interpretable scale for measuring absolute microbial load. The function automatically handles:Species or genus-level spike-in identification, Safe tree and taxonomy synchronization, Volume-based scaling (via the spiked.volume field in metadata), Optional export of intermediate results for traceability (Total_Reads.csv, Spiked_Reads.csv, Scaling_Factors.csv).

#                      CALCULATE SCALING FACTORS

result <- calculate_spikeIn_factors(
  Spiked_16S_sum_scaled_filtered,
  spiked_cells = spiked_cells,
  merged_spiked_species = species_name)

# View extracted outputs
result$spiked_species_reads # Merged spiked species name

##                              Sample Spiked_Reads
## STP1719.20422_S47 STP1719.20422_S47        14554
## STP213.20423_S59   STP213.20423_S59           83
## STP268.20424_S71   STP268.20424_S71           17
## STP544.20419_S11   STP544.20419_S11         2259
## STP570.20420_S23   STP570.20420_S23          822
## STP579.20421_S35   STP579.20421_S35         1759
## STP614.20418_S94   STP614.20418_S94            0
## UHM1000.20604_S22 UHM1000.20604_S22          118
## UHM1001.20609_S82 UHM1001.20609_S82           93
## UHM1007.20622_S48 UHM1007.20622_S48          118
## UHM1009.20614_S47 UHM1009.20614_S47          116
## UHM1010.20621_S36 UHM1010.20621_S36          979
## UHM1011.20606_S46 UHM1011.20606_S46          130
## UHM1024.20620_S24 UHM1024.20620_S24          994
## UHM1026.20607_S58 UHM1026.20607_S58          396
## UHM1028.20613_S35 UHM1028.20613_S35          898
## UHM1032.20605_S34 UHM1032.20605_S34         1616
## UHM1033.20619_S12 UHM1033.20619_S12        37416
## UHM1034.20616_S71 UHM1034.20616_S71            4
## UHM1035.20611_S11 UHM1035.20611_S11          210
## UHM1036.20612_S23 UHM1036.20612_S23          249
## UHM1052.20615_S59 UHM1052.20615_S59          150
## UHM1060.20723_S1   UHM1060.20723_S1        24617
## UHM1065.20724_S13 UHM1065.20724_S13         8179
## UHM1068.20732_S14 UHM1068.20732_S14          243
## UHM1069.20742_S39 UHM1069.20742_S39          343
## UHM1070.20725_S25 UHM1070.20725_S25         1741
## UHM1071.20733_S26 UHM1071.20733_S26           56
## UHM1072.20734_S38 UHM1072.20734_S38         1486
## UHM1073.20735_S50 UHM1073.20735_S50           39
## UHM1075.20726_S37 UHM1075.20726_S37         1179
## UHM1077.20736_S62 UHM1077.20736_S62           73
## UHM1078.20727_S49 UHM1078.20727_S49          176
## UHM1080.20737_S74 UHM1080.20737_S74           69
## UHM1081.20728_S61 UHM1081.20728_S61         1772
## UHM1088.20738_S86 UHM1088.20738_S86          240
## UHM1090.20739_S3   UHM1090.20739_S3         5303
## UHM1093.20729_S73 UHM1093.20729_S73          336
## UHM1095.20730_S85 UHM1095.20730_S85         6219
## UHM1097.20623_S60 UHM1097.20623_S60         6060
## UHM1099.20608_S70 UHM1099.20608_S70           24
## UHM1100.20788_S21 UHM1100.20788_S21           75
## UHM1102.20789_S33 UHM1102.20789_S33          108
## UHM1104.20790_S45 UHM1104.20790_S45           86
## UHM1105.20791_S57 UHM1105.20791_S57          148
## UHM1109.20531_S1   UHM1109.20531_S1          116
## UHM1110.20568_S65 UHM1110.20568_S65           50
## UHM1113.20792_S69 UHM1113.20792_S69           79
## UHM1114.20793_S81 UHM1114.20793_S81          370
## UHM1115.20794_S93 UHM1115.20794_S93          181
## UHM1117.20795_S10 UHM1117.20795_S10           61
## UHM1118.20796_S22 UHM1118.20796_S22          248
## UHM1120.20797_S34 UHM1120.20797_S34          718
## UHM1124.20798_S46 UHM1124.20798_S46          587
## UHM1126.20799_S58 UHM1126.20799_S58          196
## UHM1128.20800_S70 UHM1128.20800_S70          183
## UHM1140.20555_S4   UHM1140.20555_S4          130
## UHM1145.20801_S82 UHM1145.20801_S82           45
## UHM1163.20405_S33 UHM1163.20405_S33          150
## UHM1164.20402_S92 UHM1164.20402_S92            3
## UHM1169.20552_S63 UHM1169.20552_S63         6591
## UHM1171.20579_S7   UHM1171.20579_S7           97
## UHM1176.20404_S21 UHM1176.20404_S21           67
## UHM1177.20546_S86 UHM1177.20546_S86         1558
## UHM1182.20576_S66 UHM1182.20576_S66          233
## UHM1210.20802_S94 UHM1210.20802_S94           98
## UHM1212.20803_S11 UHM1212.20803_S11          312
## UHM1217.20804_S23 UHM1217.20804_S23          198
## UHM1218.20805_S35 UHM1218.20805_S35          151
## UHM1219.20806_S47 UHM1219.20806_S47           57
## UHM1220.20807_S59 UHM1220.20807_S59          104
## UHM1221.20808_S71 UHM1221.20808_S71          105
## UHM1222.20809_S83 UHM1222.20809_S83         3039
## UHM1223.20810_S95 UHM1223.20810_S95           69
## UHM1225.20811_S12 UHM1225.20811_S12          229
## UHM1227.20812_S24 UHM1227.20812_S24          878
## UHM1228.20813_S36 UHM1228.20813_S36           69
## UHM1237.20814_S48 UHM1237.20814_S48          413
## UHM1240.20566_S41 UHM1240.20566_S41          683
## UHM1246.20815_S60 UHM1246.20815_S60          573
## UHM1247.20816_S72 UHM1247.20816_S72          235
## UHM1248.20575_S54 UHM1248.20575_S54         3287
## UHM1256.20570_S89 UHM1256.20570_S89          901
## UHM1260.20596_S21 UHM1260.20596_S21          614
## UHM1270.20577_S78 UHM1270.20577_S78          217
## UHM1271.20397_S32 UHM1271.20397_S32          896
## UHM1272.20398_S44 UHM1272.20398_S44         1529
## UHM1274.20554_S87 UHM1274.20554_S87          467
## UHM1275.20597_S33 UHM1275.20597_S33         1203
## UHM1282.20599_S57 UHM1282.20599_S57         3820
## UHM1287.20543_S50 UHM1287.20543_S50         1979
## UHM1291.20416_S70 UHM1291.20416_S70          261
## UHM1296.20550_S39 UHM1296.20550_S39           76
## UHM1319.20561_S76 UHM1319.20561_S76         1567
## UHM1324.20413_S34 UHM1324.20413_S34          433
## UHM1327.20545_S74 UHM1327.20545_S74           73
## UHM1328.20572_S18 UHM1328.20572_S18         3350
## UHM1334.20417_S82 UHM1334.20417_S82          155
## UHM1338.20399_S56 UHM1338.20399_S56          760
## UHM1341.20602_S93 UHM1341.20602_S93         8050
## UHM1356.20541_S26 UHM1356.20541_S26          810
## UHM1380.20580_S19 UHM1380.20580_S19          308
## UHM1383.20594_S92 UHM1383.20594_S92          421
## UHM1385.20563_S5   UHM1385.20563_S5        20431
## UHM1399.20756_S17 UHM1399.20756_S17          918
## UHM1400.20757_S29 UHM1400.20757_S29          316
## UHM1401.20758_S41 UHM1401.20758_S41         2414
## UHM1402.20759_S53 UHM1402.20759_S53          166
## UHM1403.20760_S65 UHM1403.20760_S65          309
## UHM1405.20761_S77 UHM1405.20761_S77          219
## UHM1406.20762_S89 UHM1406.20762_S89          223
## UHM1414.20763_S6   UHM1414.20763_S6          107
## UHM1419.20764_S18 UHM1419.20764_S18          488
## UHM1427.20389_S31 UHM1427.20389_S31          103
## UHM1428.20390_S43 UHM1428.20390_S43            0
## UHM1429.20391_S55 UHM1429.20391_S55           29
## UHM1430.20392_S67 UHM1430.20392_S67           11
## UHM1432.20393_S79 UHM1432.20393_S79           18
## UHM1435.20388_S19 UHM1435.20388_S19            0
## UHM162.20560_S64   UHM162.20560_S64          704
## UHM198.20585_S79   UHM198.20585_S79         3649
## UHM20.3314_S52       UHM20.3314_S52           43
## UHM20.3315_S64       UHM20.3315_S64          349
## UHM204.20409_S81   UHM204.20409_S81           66
## UHM206.20410_S93   UHM206.20410_S93            0
## UHM207.20593_S80   UHM207.20593_S80          200
## UHM208.20411_S10   UHM208.20411_S10          391
## UHM211.20406_S45   UHM211.20406_S45          164
## UHM215.20408_S69   UHM215.20408_S69            9
## UHM216.20429_S36   UHM216.20429_S36           63
## UHM219.20430_S48   UHM219.20430_S48        21041
## UHM236.20431_S60   UHM236.20431_S60           41
## UHM238.20407_S57   UHM238.20407_S57          120
## UHM245.20538_S85   UHM245.20538_S85          108
## UHM252.20558_S40   UHM252.20558_S40         1090
## UHM267.20400_S68   UHM267.20400_S68          131
## UHM274.20581_S31   UHM274.20581_S31         2025
## UHM276.20586_S91   UHM276.20586_S91         1169
## UHM280.20401_S80   UHM280.20401_S80          389
## UHM286.20425_S83   UHM286.20425_S83           30
## UHM289.20426_S95   UHM289.20426_S95            4
## UHM294.20427_S12   UHM294.20427_S12          610
## UHM298.20600_S69   UHM298.20600_S69          404
## UHM325.20548_S15   UHM325.20548_S15          584
## UHM337.20412_S22   UHM337.20412_S22          261
## UHM354.20535_S49   UHM354.20535_S49          355
## UHM356.20415_S58   UHM356.20415_S58          411
## UHM369.20773_S31   UHM369.20773_S31          233
## UHM370.20774_S43   UHM370.20774_S43           88
## UHM372.20775_S55   UHM372.20775_S55           30
## UHM373.20776_S67   UHM373.20776_S67           84
## UHM374.20777_S79   UHM374.20777_S79           69
## UHM375.20778_S91   UHM375.20778_S91          716
## UHM377.20779_S8     UHM377.20779_S8          477
## UHM38.3376_S36       UHM38.3376_S36            0
## UHM386.20781_S32   UHM386.20781_S32          234
## UHM387.20782_S44   UHM387.20782_S44           58
## UHM414.20583_S55   UHM414.20583_S55           85
## UHM418.20765_S30   UHM418.20765_S30          270
## UHM422.20766_S42   UHM422.20766_S42         1147
## UHM425.20767_S54   UHM425.20767_S54          118
## UHM426.20534_S37   UHM426.20534_S37         4598
## UHM428.20544_S62   UHM428.20544_S62          454
## UHM429.20559_S52   UHM429.20559_S52         1283
## UHM435.20547_S3     UHM435.20547_S3         1718
## UHM437.20768_S66   UHM437.20768_S66           49
## UHM439.20564_S17   UHM439.20564_S17           71
## UHM44.3526_S31       UHM44.3526_S31         1281
## UHM445.20569_S77   UHM445.20569_S77           26
## UHM447.20783_S56   UHM447.20783_S56           64
## UHM448.20769_S78   UHM448.20769_S78          424
## UHM45.3539_S92       UHM45.3539_S92            0
## UHM454.20770_S90   UHM454.20770_S90          278
## UHM455.20785_S80   UHM455.20785_S80          792
## UHM458.20786_S92   UHM458.20786_S92           69
## UHM459.20787_S9     UHM459.20787_S9          120
## UHM461.20771_S7     UHM461.20771_S7          242
## UHM467.20772_S19   UHM467.20772_S19           47
## UHM470.20533_S25   UHM470.20533_S25         1220
## UHM476.20414_S46   UHM476.20414_S46         1138
## UHM478.20549_S27   UHM478.20549_S27          429
## UHM479.20551_S51   UHM479.20551_S51        14439
## UHM481.20403_S9     UHM481.20403_S9          227
## UHM482.20590_S44   UHM482.20590_S44          233
## UHM483.20603_S10   UHM483.20603_S10           91
## UHM519.20582_S43   UHM519.20582_S43          136
## UHM520.20573_S30   UHM520.20573_S30          253
## UHM746.21478_S117 UHM746.21478_S117        85168
## UHM747.21477_S106 UHM747.21477_S106        82287
## UHM748.21467_S170 UHM748.21467_S170        55291
## UHM748.21487_S129 UHM748.21487_S129        36462
## UHM749.21479_S128 UHM749.21479_S128        62780
## UHM759.21466_S159 UHM759.21466_S159        76705
## UHM759.21486_S118 UHM759.21486_S118        16500
## UHM775.21485_S107 UHM775.21485_S107        43046
## UHM776.21482_S161 UHM776.21482_S161        58165
## UHM777.21484_S183 UHM777.21484_S183        23485
## UHM779.21468_S181 UHM779.21468_S181        80976
## UHM779.21488_S140 UHM779.21488_S140            9
## UHM782.21480_S139 UHM782.21480_S139        38354
## UHM810.21472_S138 UHM810.21472_S138        94712
## UHM811.21471_S127 UHM811.21471_S127        30104
## UHM813.21481_S150 UHM813.21481_S150        64962
## UHM818.21469_S105 UHM818.21469_S105        50417
## UHM818.21489_S151 UHM818.21489_S151           19
## UHM819.21473_S149 UHM819.21473_S149        83424
## UHM820.21470_S116 UHM820.21470_S116        66075
## UHM820.21490_S162 UHM820.21490_S162          515
## UHM827.21474_S160 UHM827.21474_S160        83467
## UHM829.21476_S182 UHM829.21476_S182        69357
## UHM832.21483_S172 UHM832.21483_S172        68494
## UHM836.20385_S78   UHM836.20385_S78            0
## UHM837.20386_S90   UHM837.20386_S90            0
## UHM838.20387_S7     UHM838.20387_S7           11
## UHM891.20384_S66   UHM891.20384_S66           85
## UHM892.20532_S13   UHM892.20532_S13         1164
## UHM893.20595_S9     UHM893.20595_S9           16
## UHM894.20540_S14   UHM894.20540_S14           36
## UHM895.20536_S61   UHM895.20536_S61         2116
## UHM896.20601_S81   UHM896.20601_S81            0
## UHM897.20591_S56   UHM897.20591_S56            0
## UHM898.20394_S91   UHM898.20394_S91            0
## UHM899.20588_S20   UHM899.20588_S20           24
## UHM900.20395_S8     UHM900.20395_S8            0
## UHM901.20542_S38   UHM901.20542_S38           95
## UHM902.20584_S67   UHM902.20584_S67         1217
## UHM903.20587_S8     UHM903.20587_S8          470
## UHM904.20567_S53   UHM904.20567_S53           45
## UHM905.20598_S45   UHM905.20598_S45           89
## UHM906.20565_S29   UHM906.20565_S29            0
## UHM907.20592_S68   UHM907.20592_S68           30
## UHM908.20396_S20   UHM908.20396_S20            0
## UHM909.20557_S28   UHM909.20557_S28           87
## UHM910.20562_S88   UHM910.20562_S88           34
## UHM965.20537_S73   UHM965.20537_S73          464
## UHM966.20743_S51   UHM966.20743_S51          461
## UHM967.20744_S63   UHM967.20744_S63            0
## UHM968.20571_S6     UHM968.20571_S6        33012
## UHM969.20745_S75   UHM969.20745_S75           30
## UHM971.20746_S87   UHM971.20746_S87           10
## UHM973.20578_S90   UHM973.20578_S90           46
## UHM974.20432_S72   UHM974.20432_S72            0
## UHM975.20747_S4     UHM975.20747_S4           33
## UHM977.20748_S16   UHM977.20748_S16          100
## UHM978.20749_S28   UHM978.20749_S28           91
## UHM979.20750_S40   UHM979.20750_S40           35
## UHM980.20731_S2     UHM980.20731_S2          248
## UHM981.20539_S2     UHM981.20539_S2          766
## UHM982.20740_S15   UHM982.20740_S15          161
## UHM983.20556_S16   UHM983.20556_S16        12547
## UHM984.20751_S52   UHM984.20751_S52           26
## UHM985.20752_S64   UHM985.20752_S64          801
## UHM988.20753_S76   UHM988.20753_S76          603
## UHM989.20754_S88   UHM989.20754_S88           17
## UHM991.20755_S5     UHM991.20755_S5         1868
## UHM993.20741_S27   UHM993.20741_S27           52
## UHM996.20610_S94   UHM996.20610_S94            0
## UHM997.20553_S75   UHM997.20553_S75            0
## UHM998.20618_S95   UHM998.20618_S95         2610
## UHM999.20617_S83   UHM999.20617_S83           72

result$total_reads # Total reads detected for the spike

##                              Sample Total_Reads
## STP1719.20422_S47 STP1719.20422_S47       19142
## STP213.20423_S59   STP213.20423_S59        8462
## STP268.20424_S71   STP268.20424_S71        6968
## STP544.20419_S11   STP544.20419_S11        2340
## STP570.20420_S23   STP570.20420_S23        2647
## STP579.20421_S35   STP579.20421_S35        5193
## STP614.20418_S94   STP614.20418_S94           5
## UHM1000.20604_S22 UHM1000.20604_S22        5538
## UHM1001.20609_S82 UHM1001.20609_S82        3957
## UHM1007.20622_S48 UHM1007.20622_S48       11042
## UHM1009.20614_S47 UHM1009.20614_S47       22828
## UHM1010.20621_S36 UHM1010.20621_S36        6453
## UHM1011.20606_S46 UHM1011.20606_S46       11406
## UHM1024.20620_S24 UHM1024.20620_S24        2889
## UHM1026.20607_S58 UHM1026.20607_S58        3207
## UHM1028.20613_S35 UHM1028.20613_S35        2555
## UHM1032.20605_S34 UHM1032.20605_S34        2472
## UHM1033.20619_S12 UHM1033.20619_S12       39314
## UHM1034.20616_S71 UHM1034.20616_S71        3389
## UHM1035.20611_S11 UHM1035.20611_S11        1837
## UHM1036.20612_S23 UHM1036.20612_S23        2327
## UHM1052.20615_S59 UHM1052.20615_S59       16010
## UHM1060.20723_S1   UHM1060.20723_S1       27549
## UHM1065.20724_S13 UHM1065.20724_S13       10557
## UHM1068.20732_S14 UHM1068.20732_S14        2526
## UHM1069.20742_S39 UHM1069.20742_S39       16428
## UHM1070.20725_S25 UHM1070.20725_S25       10491
## UHM1071.20733_S26 UHM1071.20733_S26        2028
## UHM1072.20734_S38 UHM1072.20734_S38        3425
## UHM1073.20735_S50 UHM1073.20735_S50        2731
## UHM1075.20726_S37 UHM1075.20726_S37        7732
## UHM1077.20736_S62 UHM1077.20736_S62        9489
## UHM1078.20727_S49 UHM1078.20727_S49        4532
## UHM1080.20737_S74 UHM1080.20737_S74        8886
## UHM1081.20728_S61 UHM1081.20728_S61        2033
## UHM1088.20738_S86 UHM1088.20738_S86        6274
## UHM1090.20739_S3   UHM1090.20739_S3       12258
## UHM1093.20729_S73 UHM1093.20729_S73        2009
## UHM1095.20730_S85 UHM1095.20730_S85       11680
## UHM1097.20623_S60 UHM1097.20623_S60       10958
## UHM1099.20608_S70 UHM1099.20608_S70        3670
## UHM1100.20788_S21 UHM1100.20788_S21        4879
## UHM1102.20789_S33 UHM1102.20789_S33        2410
## UHM1104.20790_S45 UHM1104.20790_S45        3410
## UHM1105.20791_S57 UHM1105.20791_S57        2360
## UHM1109.20531_S1   UHM1109.20531_S1        3122
## UHM1110.20568_S65 UHM1110.20568_S65        6965
## UHM1113.20792_S69 UHM1113.20792_S69       16025
## UHM1114.20793_S81 UHM1114.20793_S81        5041
## UHM1115.20794_S93 UHM1115.20794_S93        5256
## UHM1117.20795_S10 UHM1117.20795_S10        3976
## UHM1118.20796_S22 UHM1118.20796_S22        5348
## UHM1120.20797_S34 UHM1120.20797_S34        2506
## UHM1124.20798_S46 UHM1124.20798_S46        6294
## UHM1126.20799_S58 UHM1126.20799_S58        5941
## UHM1128.20800_S70 UHM1128.20800_S70        2595
## UHM1140.20555_S4   UHM1140.20555_S4       10197
## UHM1145.20801_S82 UHM1145.20801_S82        3319
## UHM1163.20405_S33 UHM1163.20405_S33        5402
## UHM1164.20402_S92 UHM1164.20402_S92         911
## UHM1169.20552_S63 UHM1169.20552_S63        7438
## UHM1171.20579_S7   UHM1171.20579_S7        4952
## UHM1176.20404_S21 UHM1176.20404_S21        5583
## UHM1177.20546_S86 UHM1177.20546_S86        5901
## UHM1182.20576_S66 UHM1182.20576_S66        2437
## UHM1210.20802_S94 UHM1210.20802_S94        5710
## UHM1212.20803_S11 UHM1212.20803_S11        5886
## UHM1217.20804_S23 UHM1217.20804_S23        6804
## UHM1218.20805_S35 UHM1218.20805_S35        3493
## UHM1219.20806_S47 UHM1219.20806_S47        2276
## UHM1220.20807_S59 UHM1220.20807_S59        3906
## UHM1221.20808_S71 UHM1221.20808_S71        2137
## UHM1222.20809_S83 UHM1222.20809_S83        8953
## UHM1223.20810_S95 UHM1223.20810_S95        4269
## UHM1225.20811_S12 UHM1225.20811_S12        5986
## UHM1227.20812_S24 UHM1227.20812_S24        9891
## UHM1228.20813_S36 UHM1228.20813_S36        5503
## UHM1237.20814_S48 UHM1237.20814_S48        5591
## UHM1240.20566_S41 UHM1240.20566_S41       12987
## UHM1246.20815_S60 UHM1246.20815_S60        5004
## UHM1247.20816_S72 UHM1247.20816_S72        3378
## UHM1248.20575_S54 UHM1248.20575_S54       13609
## UHM1256.20570_S89 UHM1256.20570_S89       17303
## UHM1260.20596_S21 UHM1260.20596_S21       11405
## UHM1270.20577_S78 UHM1270.20577_S78        1427
## UHM1271.20397_S32 UHM1271.20397_S32        9272
## UHM1272.20398_S44 UHM1272.20398_S44        8652
## UHM1274.20554_S87 UHM1274.20554_S87        8200
## UHM1275.20597_S33 UHM1275.20597_S33        9805
## UHM1282.20599_S57 UHM1282.20599_S57       13538
## UHM1287.20543_S50 UHM1287.20543_S50       11499
## UHM1291.20416_S70 UHM1291.20416_S70       10486
## UHM1296.20550_S39 UHM1296.20550_S39        8543
## UHM1319.20561_S76 UHM1319.20561_S76        9848
## UHM1324.20413_S34 UHM1324.20413_S34       10595
## UHM1327.20545_S74 UHM1327.20545_S74       10782
## UHM1328.20572_S18 UHM1328.20572_S18       16676
## UHM1334.20417_S82 UHM1334.20417_S82        6421
## UHM1338.20399_S56 UHM1338.20399_S56       10825
## UHM1341.20602_S93 UHM1341.20602_S93       21036
## UHM1356.20541_S26 UHM1356.20541_S26        9245
## UHM1380.20580_S19 UHM1380.20580_S19       11947
## UHM1383.20594_S92 UHM1383.20594_S92        6453
## UHM1385.20563_S5   UHM1385.20563_S5       24006
## UHM1399.20756_S17 UHM1399.20756_S17        5659
## UHM1400.20757_S29 UHM1400.20757_S29        8433
## UHM1401.20758_S41 UHM1401.20758_S41        6518
## UHM1402.20759_S53 UHM1402.20759_S53        7091
## UHM1403.20760_S65 UHM1403.20760_S65        7923
## UHM1405.20761_S77 UHM1405.20761_S77        5923
## UHM1406.20762_S89 UHM1406.20762_S89       10431
## UHM1414.20763_S6   UHM1414.20763_S6        6573
## UHM1419.20764_S18 UHM1419.20764_S18        5863
## UHM1427.20389_S31 UHM1427.20389_S31        5228
## UHM1428.20390_S43 UHM1428.20390_S43        4532
## UHM1429.20391_S55 UHM1429.20391_S55        3859
## UHM1430.20392_S67 UHM1430.20392_S67        5040
## UHM1432.20393_S79 UHM1432.20393_S79        4047
## UHM1435.20388_S19 UHM1435.20388_S19        9292
## UHM162.20560_S64   UHM162.20560_S64       14141
## UHM198.20585_S79   UHM198.20585_S79       11058
## UHM20.3314_S52       UHM20.3314_S52       31095
## UHM20.3315_S64       UHM20.3315_S64       66533
## UHM204.20409_S81   UHM204.20409_S81        5723
## UHM206.20410_S93   UHM206.20410_S93         523
## UHM207.20593_S80   UHM207.20593_S80        8319
## UHM208.20411_S10   UHM208.20411_S10        7217
## UHM211.20406_S45   UHM211.20406_S45        7727
## UHM215.20408_S69   UHM215.20408_S69       19236
## UHM216.20429_S36   UHM216.20429_S36        7081
## UHM219.20430_S48   UHM219.20430_S48       21661
## UHM236.20431_S60   UHM236.20431_S60        4428
## UHM238.20407_S57   UHM238.20407_S57        4161
## UHM245.20538_S85   UHM245.20538_S85       32445
## UHM252.20558_S40   UHM252.20558_S40        8412
## UHM267.20400_S68   UHM267.20400_S68       12606
## UHM274.20581_S31   UHM274.20581_S31       15008
## UHM276.20586_S91   UHM276.20586_S91       13645
## UHM280.20401_S80   UHM280.20401_S80        7442
## UHM286.20425_S83   UHM286.20425_S83        2127
## UHM289.20426_S95   UHM289.20426_S95          13
## UHM294.20427_S12   UHM294.20427_S12        5674
## UHM298.20600_S69   UHM298.20600_S69       22131
## UHM325.20548_S15   UHM325.20548_S15        7768
## UHM337.20412_S22   UHM337.20412_S22        4612
## UHM354.20535_S49   UHM354.20535_S49       11303
## UHM356.20415_S58   UHM356.20415_S58        9725
## UHM369.20773_S31   UHM369.20773_S31       10033
## UHM370.20774_S43   UHM370.20774_S43        4214
## UHM372.20775_S55   UHM372.20775_S55         343
## UHM373.20776_S67   UHM373.20776_S67        5140
## UHM374.20777_S79   UHM374.20777_S79        3588
## UHM375.20778_S91   UHM375.20778_S91        7304
## UHM377.20779_S8     UHM377.20779_S8        7549
## UHM38.3376_S36       UHM38.3376_S36       29112
## UHM386.20781_S32   UHM386.20781_S32        6867
## UHM387.20782_S44   UHM387.20782_S44        2149
## UHM414.20583_S55   UHM414.20583_S55       12802
## UHM418.20765_S30   UHM418.20765_S30        6818
## UHM422.20766_S42   UHM422.20766_S42        6452
## UHM425.20767_S54   UHM425.20767_S54        4230
## UHM426.20534_S37   UHM426.20534_S37       13508
## UHM428.20544_S62   UHM428.20544_S62        3325
## UHM429.20559_S52   UHM429.20559_S52       14735
## UHM435.20547_S3     UHM435.20547_S3       12442
## UHM437.20768_S66   UHM437.20768_S66        3931
## UHM439.20564_S17   UHM439.20564_S17        3706
## UHM44.3526_S31       UHM44.3526_S31        3822
## UHM445.20569_S77   UHM445.20569_S77        1267
## UHM447.20783_S56   UHM447.20783_S56        5833
## UHM448.20769_S78   UHM448.20769_S78        6456
## UHM45.3539_S92       UHM45.3539_S92       35221
## UHM454.20770_S90   UHM454.20770_S90        6567
## UHM455.20785_S80   UHM455.20785_S80        6400
## UHM458.20786_S92   UHM458.20786_S92        3554
## UHM459.20787_S9     UHM459.20787_S9        6949
## UHM461.20771_S7     UHM461.20771_S7       10174
## UHM467.20772_S19   UHM467.20772_S19        4970
## UHM470.20533_S25   UHM470.20533_S25        6080
## UHM476.20414_S46   UHM476.20414_S46       12376
## UHM478.20549_S27   UHM478.20549_S27        6455
## UHM479.20551_S51   UHM479.20551_S51       16542
## UHM481.20403_S9     UHM481.20403_S9        1938
## UHM482.20590_S44   UHM482.20590_S44         254
## UHM483.20603_S10   UHM483.20603_S10        4453
## UHM519.20582_S43   UHM519.20582_S43       16240
## UHM520.20573_S30   UHM520.20573_S30        9366
## UHM746.21478_S117 UHM746.21478_S117       85545
## UHM747.21477_S106 UHM747.21477_S106       82434
## UHM748.21467_S170 UHM748.21467_S170       55338
## UHM748.21487_S129 UHM748.21487_S129       36665
## UHM749.21479_S128 UHM749.21479_S128       62924
## UHM759.21466_S159 UHM759.21466_S159       76912
## UHM759.21486_S118 UHM759.21486_S118       16768
## UHM775.21485_S107 UHM775.21485_S107       43137
## UHM776.21482_S161 UHM776.21482_S161       58259
## UHM777.21484_S183 UHM777.21484_S183       23552
## UHM779.21468_S181 UHM779.21468_S181       81397
## UHM779.21488_S140 UHM779.21488_S140           9
## UHM782.21480_S139 UHM782.21480_S139       38391
## UHM810.21472_S138 UHM810.21472_S138       94869
## UHM811.21471_S127 UHM811.21471_S127       30255
## UHM813.21481_S150 UHM813.21481_S150       65073
## UHM818.21469_S105 UHM818.21469_S105       50515
## UHM818.21489_S151 UHM818.21489_S151          30
## UHM819.21473_S149 UHM819.21473_S149       83507
## UHM820.21470_S116 UHM820.21470_S116       66154
## UHM820.21490_S162 UHM820.21490_S162         518
## UHM827.21474_S160 UHM827.21474_S160       84834
## UHM829.21476_S182 UHM829.21476_S182       69806
## UHM832.21483_S172 UHM832.21483_S172       76349
## UHM836.20385_S78   UHM836.20385_S78        3026
## UHM837.20386_S90   UHM837.20386_S90        2442
## UHM838.20387_S7     UHM838.20387_S7        4330
## UHM891.20384_S66   UHM891.20384_S66        3238
## UHM892.20532_S13   UHM892.20532_S13        7052
## UHM893.20595_S9     UHM893.20595_S9        9991
## UHM894.20540_S14   UHM894.20540_S14        5678
## UHM895.20536_S61   UHM895.20536_S61        6765
## UHM896.20601_S81   UHM896.20601_S81       10057
## UHM897.20591_S56   UHM897.20591_S56       13724
## UHM898.20394_S91   UHM898.20394_S91         603
## UHM899.20588_S20   UHM899.20588_S20       10593
## UHM900.20395_S8     UHM900.20395_S8        6596
## UHM901.20542_S38   UHM901.20542_S38       12693
## UHM902.20584_S67   UHM902.20584_S67        4178
## UHM903.20587_S8     UHM903.20587_S8        4707
## UHM904.20567_S53   UHM904.20567_S53       16165
## UHM905.20598_S45   UHM905.20598_S45        3207
## UHM906.20565_S29   UHM906.20565_S29       11360
## UHM907.20592_S68   UHM907.20592_S68        6155
## UHM908.20396_S20   UHM908.20396_S20        8212
## UHM909.20557_S28   UHM909.20557_S28        2759
## UHM910.20562_S88   UHM910.20562_S88        6385
## UHM965.20537_S73   UHM965.20537_S73        1777
## UHM966.20743_S51   UHM966.20743_S51        7100
## UHM967.20744_S63   UHM967.20744_S63        4222
## UHM968.20571_S6     UHM968.20571_S6       33909
## UHM969.20745_S75   UHM969.20745_S75        6115
## UHM971.20746_S87   UHM971.20746_S87        6911
## UHM973.20578_S90   UHM973.20578_S90        2680
## UHM974.20432_S72   UHM974.20432_S72        1133
## UHM975.20747_S4     UHM975.20747_S4        6185
## UHM977.20748_S16   UHM977.20748_S16        7885
## UHM978.20749_S28   UHM978.20749_S28        6336
## UHM979.20750_S40   UHM979.20750_S40        5449
## UHM980.20731_S2     UHM980.20731_S2        3823
## UHM981.20539_S2     UHM981.20539_S2       16669
## UHM982.20740_S15   UHM982.20740_S15        5740
## UHM983.20556_S16   UHM983.20556_S16       12838
## UHM984.20751_S52   UHM984.20751_S52        4646
## UHM985.20752_S64   UHM985.20752_S64        2573
## UHM988.20753_S76   UHM988.20753_S76        7752
## UHM989.20754_S88   UHM989.20754_S88        5843
## UHM991.20755_S5     UHM991.20755_S5        8388
## UHM993.20741_S27   UHM993.20741_S27        6685
## UHM996.20610_S94   UHM996.20610_S94       14720
## UHM997.20553_S75   UHM997.20553_S75       12274
## UHM998.20618_S95   UHM998.20618_S95       16875
## UHM999.20617_S83   UHM999.20617_S83        7227

scaling_factors <- result$scaling_factors
head(scaling_factors) # Vector of scaling factors per sample

## STP1719.20422_S47  STP213.20423_S59  STP268.20424_S71  STP544.20419_S11 
##         0.1269067        22.2530120       108.6470588         0.8176184 
##  STP570.20420_S23  STP579.20421_S35 
##         2.2469586         1.0500284

Convert relative counts to absolute counts

This function translates relative abundance profiles into biologically meaningful absolute counts by applying empirically derived scaling factors. Absolute quantification enables direct comparison of microbial loads across samples and treatments, avoiding compositional artifacts inherent to relative data. This transformation is fundamental for integrative multi-omics analyses, network inference, and ecological interpretation of microbiome dynamics.

#             Convert relative counts to absolute counts


# absolute counts=relative countssample-specific scaling factor

# Convert to absolute counts
library(DspikeIn)
absolute <- convert_to_absolute_counts(Spiked_16S_sum_scaled_filtered, scaling_factors)

# Extract processed data
absolute_counts <- absolute$absolute_counts
tse_absolute <- absolute$obj_adj

tse_absolute <- tidy_phyloseq_tse(tse_absolute)

# View absolute count data
head(tse_absolute)

## class: TreeSummarizedExperiment 
## dim: 6 260 
## metadata(0):
## assays(1): counts
## rownames(6): OTU1 OTU2 ... OTU5 OTU6
## rowData names(7): Kingdom Phylum ... Genus Species
## colnames(260): STP1719.20422_S47 STP213.20423_S59 ... UHM998.20618_S95
##   UHM999.20617_S83
## colData names(34): sample.id X16S.biosample ... swab.presence MK.spike
## reducedDimNames(0):
## mainExpName: NULL
## altExpNames(0):
## rowLinks: a LinkDataFrame (6 rows)
## rowTree: 1 phylo tree(s) (9227 leaves)
## colLinks: NULL
## colTree: NULL

Summary Stat

This function generates a concise statistical summary of numeric variables, including measures of central tendency (mean, median), dispersion (standard deviation, standard error), and distribution (quartiles). The results can be exported as both .docx and .csv files for reporting.

#                      CALCULATE SPIKE PERCENTAGE & summary stat

# ** Calculate spike percentage & Generate summary statistics for absolute counts**
# absolute_counts is a data.frame of OTU/ASV table
# Generate summary statistics for absolute counts
post_eval_summary <- calculate_summary_stats_table(absolute_counts)

# You may want to Back normal to check calculation accuracy
# the scaling factor was computed based on spiked species reads and fixed cell count.
# Multiplying the spiked species read count by this scaling factor restores the exact spiked cell count.
# lets check it
# BackNormal <- calculate_spike_percentage(
#  tse_absolute,
#  merged_spiked_species,
#  passed_range = c(0.1, 20))

Conclusion

conclusion() evaluates spike-in performance and recovery, i.e., how well the spike-in behaved after normalization and how consistent it is across samples. The function relies on outputs from calculate_spike_percentage() and indirectly reflects how scaling factors and normalization affected your dataset. It is adviced to be performed after absolute abundance conversion because -> If you summarize spike-in performance before normalization, youre essentially measuring how biased your workflow was, not how successfully it was corrected.

# * Calculate the percentage of spiked species retrieval per sample*

library(mia)         
library(dplyr)
library(SummarizedExperiment)

# Subset TSE to remove blanks 
tse_absolute_filtered <- tse_absolute[, colData(tse_absolute)$sample.or.blank != "blank"]

# Generate conclusion report
conc <- conclusion(
  tse_absolute_filtered,
  merged_spiked_species,
  max_passed_range = 20,
  output_path = output_path)

conc$full_report

##                Sample Total_Reads Spiked_Reads   Percentage Result
## 1   STP1719.20422_S47        2427         1847  76.10218377 failed
## 2    STP213.20423_S59      188314         1847   0.98080865 passed
## 3    STP268.20424_S71      757053         1847   0.24397235 passed
## 4    STP544.20419_S11        1913         1847  96.54992159 failed
## 5    STP570.20420_S23        5948         1847  31.05245461 failed
## 6    STP579.20421_S35        5452         1847  33.87747616 failed
## 7    STP614.20418_S94           5            0   0.00000000 failed
## 8   UHM1000.20604_S22       43347          924   2.13163541 passed
## 9   UHM1001.20609_S82       39309          924   2.35060673 passed
## 10  UHM1007.20622_S48       86418          924   1.06922169 passed
## 11  UHM1009.20614_S47      181742          924   0.50841303 passed
## 12  UHM1010.20621_S36        6123          924  15.09064184 passed
## 13  UHM1011.20606_S46       81017          924   1.14050138 passed
## 14  UHM1024.20620_S24        2690          924  34.34944238 failed
## 15  UHM1026.20607_S58        7494          924  12.32986389 passed
## 16  UHM1028.20613_S35        2620          924  35.26717557 failed
## 17  UHM1032.20605_S34        1410          924  65.53191489 failed
## 18  UHM1033.20619_S12         966          924  95.65217391 failed
## 19  UHM1034.20616_S71      782439          924   0.11809227 passed
## 20  UHM1035.20611_S11        8082          924  11.43281366 passed
## 21  UHM1036.20612_S23        8625          924  10.71304348 passed
## 22  UHM1052.20615_S59       98557          924   0.93752854 passed
## 23   UHM1060.20723_S1        2062         1847  89.57322987 failed
## 24  UHM1065.20724_S13        2384         1847  77.47483221 failed
## 25  UHM1068.20732_S14       19198         1847   9.62079383 passed
## 26  UHM1069.20742_S39       88462         1847   2.08790215 passed
## 27  UHM1070.20725_S25       11119         1847  16.61120604 passed
## 28  UHM1071.20733_S26       66892         1847   2.76116725 passed
## 29  UHM1072.20734_S38        4254         1847  43.41795957 failed
## 30  UHM1073.20735_S50      129333         1847   1.42809646 passed
## 31  UHM1075.20726_S37       12116         1847  15.24430505 passed
## 32  UHM1077.20736_S62      240090         1847   0.76929485 passed
## 33  UHM1078.20727_S49       47549         1847   3.88441397 passed
## 34  UHM1080.20737_S74      237852         1847   0.77653331 passed
## 35  UHM1081.20728_S61        2118         1847  87.20491029 failed
## 36  UHM1088.20738_S86       48261         1847   3.82710677 passed
## 37   UHM1090.20739_S3        4260         1847  43.35680751 failed
## 38  UHM1093.20729_S73       11040         1847  16.73007246 passed
## 39  UHM1095.20730_S85        3475         1847  53.15107914 failed
## 40  UHM1097.20623_S60        1639          924  56.37583893 failed
## 41  UHM1099.20608_S70      141219          924   0.65430289 passed
## 42  UHM1100.20788_S21      120156         1847   1.53716835 passed
## 43  UHM1102.20789_S33       41207         1847   4.48224816 passed
## 44  UHM1104.20790_S45       73225         1847   2.52236258 passed
## 45  UHM1105.20791_S57       29445         1847   6.27271184 passed
## 46   UHM1109.20531_S1       49710         1847   3.71555019 passed
## 47  UHM1110.20568_S65      257312         1847   0.71780562 passed
## 48  UHM1113.20792_S69      374668         1847   0.49296978 passed
## 49  UHM1114.20793_S81       25164         1847   7.33985058 passed
## 50  UHM1115.20794_S93       53637         1847   3.44351847 passed
## 51  UHM1117.20795_S10      120412         1847   1.53390028 passed
## 52  UHM1118.20796_S22       39826         1847   4.63767388 passed
## 53  UHM1120.20797_S34        6447         1847  28.64898402 failed
## 54  UHM1124.20798_S46       19768         1847   9.34338325 passed
## 55  UHM1126.20799_S58       55979         1847   3.29945158 passed
## 56  UHM1128.20800_S70       26179         1847   7.05527331 passed
## 57   UHM1140.20555_S4      144876         1847   1.27488335 passed
## 58  UHM1145.20801_S82      136223         1847   1.35586502 passed
## 59  UHM1163.20405_S33       66525         1847   2.77639985 passed
## 60  UHM1164.20402_S92      560873         1847   0.32930806 passed
## 61  UHM1169.20552_S63        2087         1847  88.50023958 failed
## 62   UHM1171.20579_S7       94275         1847   1.95916203 passed
## 63  UHM1176.20404_S21      153911         1847   1.20004418 passed
## 64  UHM1177.20546_S86        7001         1847  26.38194544 failed
## 65  UHM1182.20576_S66       19322         1847   9.55905186 passed
## 66  UHM1210.20802_S94      107631         1847   1.71604835 passed
## 67  UHM1212.20803_S11       34861         1847   5.29818422 passed
## 68  UHM1217.20804_S23       63472         1847   2.90994454 passed
## 69  UHM1218.20805_S35       42728         1847   4.32269238 passed
## 70  UHM1219.20806_S47       73751         1847   2.50437282 passed
## 71  UHM1220.20807_S59       69371         1847   2.66249586 passed
## 72  UHM1221.20808_S71       37595         1847   4.91288735 passed
## 73  UHM1222.20809_S83        5442         1847  33.93972804 failed
## 74  UHM1223.20810_S95      114277         1847   1.61624824 passed
## 75  UHM1225.20811_S12       48275         1847   3.82599689 passed
## 76  UHM1227.20812_S24       20802         1847   8.87895395 passed
## 77  UHM1228.20813_S36      147297         1847   1.25392914 passed
## 78  UHM1237.20814_S48       24999         1847   7.38829553 passed
## 79  UHM1240.20566_S41       35116         1847   5.25971067 passed
## 80  UHM1246.20815_S60       16128         1847  11.45213294 passed
## 81  UHM1247.20816_S72       26551         1847   6.95642349 passed
## 82  UHM1248.20575_S54        7646         1847  24.15642166 failed
## 83  UHM1256.20570_S89       35459         1847   5.20883274 passed
## 84  UHM1260.20596_S21       34301         1847   5.38468266 passed
## 85  UHM1270.20577_S78       12149         1847  15.20289736 passed
## 86  UHM1271.20397_S32       19107         1847   9.66661433 passed
## 87  UHM1272.20398_S44       10447         1847  17.67971667 passed
## 88  UHM1274.20554_S87       32434         1847   5.69464143 passed
## 89  UHM1275.20597_S33       15061         1847  12.26346192 passed
## 90  UHM1282.20599_S57        6536         1847  28.25887393 failed
## 91  UHM1287.20543_S50       10735         1847  17.20540289 passed
## 92  UHM1291.20416_S70       74189         1847   2.48958741 passed
## 93  UHM1296.20550_S39      207620         1847   0.88960601 passed
## 94  UHM1319.20561_S76       11611         1847  15.90732926 passed
## 95  UHM1324.20413_S34       45204         1847   4.08592160 passed
## 96  UHM1327.20545_S74      272806         1847   0.67703790 passed
## 97  UHM1328.20572_S18        9198         1847  20.08045227 failed
## 98  UHM1334.20417_S82       76515         1847   2.41390577 passed
## 99  UHM1338.20399_S56       26305         1847   7.02147881 passed
## 100 UHM1341.20602_S93        4829         1847  38.24808449 failed
## 101 UHM1356.20541_S26       21082         1847   8.76102837 passed
## 102 UHM1380.20580_S19       71651         1847   2.57777282 passed
## 103 UHM1383.20594_S92       28316         1847   6.52281396 passed
## 104  UHM1385.20563_S5        2166         1847  85.27239151 failed
## 105 UHM1399.20756_S17       11379         1847  16.23165480 passed
## 106 UHM1400.20757_S29       49300         1847   3.74645030 passed
## 107 UHM1401.20758_S41        4984         1847  37.05858748 failed
## 108 UHM1402.20759_S53       78898         1847   2.34099724 passed
## 109 UHM1403.20760_S65       47369         1847   3.89917457 passed
## 110 UHM1405.20761_S77       49948         1847   3.69784576 passed
## 111 UHM1406.20762_S89       86410         1847   2.13748409 passed
## 112  UHM1414.20763_S6      113476         1847   1.62765695 passed
## 113 UHM1419.20764_S18       22182         1847   8.32657109 passed
## 114 UHM1427.20389_S31       93753         1847   1.97007029 passed
## 115 UHM1428.20390_S43        4532            0   0.00000000 failed
## 116 UHM1429.20391_S55      245778         1847   0.75149118 passed
## 117 UHM1430.20392_S67      846266         1847   0.21825289 passed
## 118 UHM1432.20393_S79      415264         1847   0.44477730 passed
## 119 UHM1435.20388_S19        9292            0   0.00000000 failed
## 120  UHM162.20560_S64       37088         1847   4.98004745 passed
## 121  UHM198.20585_S79        5610         1847  32.92335116 failed
## 122    UHM20.3314_S52     1335639         1847   0.13828587 passed
## 123    UHM20.3315_S64      352109         1847   0.52455348 passed
## 124  UHM204.20409_S81      160159         1847   1.15322898 passed
## 125  UHM206.20410_S93         523            0   0.00000000 failed
## 126  UHM207.20593_S80       76828         1847   2.40407143 passed
## 127  UHM208.20411_S10       34089         1847   5.41817008 passed
## 128  UHM211.20406_S45       87029         1847   2.12228108 passed
## 129  UHM215.20408_S69     3947656         1847   0.04678726 failed
## 130  UHM216.20429_S36      207601         1847   0.88968743 passed
## 131  UHM219.20430_S48        1897         1847  97.36425936 failed
## 132  UHM236.20431_S60      199471         1847   0.92594914 passed
## 133  UHM238.20407_S57       64050         1847   2.88368462 passed
## 134  UHM245.20538_S85      554868         1847   0.33287196 passed
## 135  UHM252.20558_S40       14250         1847  12.96140351 passed
## 136  UHM267.20400_S68      177722         1847   1.03926357 passed
## 137  UHM274.20581_S31       13695         1847  13.48667397 passed
## 138  UHM276.20586_S91       21560         1847   8.56679035 passed
## 139  UHM280.20401_S80       35333         1847   5.22740781 passed
## 140  UHM286.20425_S83      130953         1847   1.41042970 passed
## 141  UHM289.20426_S95        6003         1847  30.76794936 failed
## 142  UHM294.20427_S12       17177         1847  10.75275077 passed
## 143  UHM298.20600_S69      101173         1847   1.82558588 passed
## 144  UHM325.20548_S15       24562         1847   7.51974595 passed
## 145  UHM337.20412_S22       32632         1847   5.66008826 passed
## 146  UHM354.20535_S49       58805         1847   3.14088938 passed
## 147  UHM356.20415_S58       43688         1847   4.22770555 passed
## 148  UHM369.20773_S31       79551         1847   2.32178100 passed
## 149  UHM370.20774_S43       88455         1847   2.08806738 passed
## 150  UHM372.20775_S55       21119         1847   8.74567925 passed
## 151  UHM373.20776_S67      113026         1847   1.63413728 passed
## 152  UHM374.20777_S79       96042         1847   1.92311697 passed
## 153  UHM375.20778_S91       18842         1847   9.80256873 passed
## 154   UHM377.20779_S8       29234         1847   6.31798591 passed
## 155    UHM38.3376_S36       29112            0   0.00000000 failed
## 156  UHM386.20781_S32       54216         1847   3.40674340 passed
## 157  UHM387.20782_S44       68442         1847   2.69863534 passed
## 158  UHM414.20583_S55      278181         1847   0.66395620 passed
## 159  UHM418.20765_S30       46656         1847   3.95876200 passed
## 160  UHM422.20766_S42       10385         1847  17.78526721 passed
## 161  UHM425.20767_S54       66208         1847   2.78969309 passed
## 162  UHM426.20534_S37        5428         1847  34.02726603 failed
## 163  UHM428.20544_S62       13527         1847  13.65417314 passed
## 164  UHM429.20559_S52       21213         1847   8.70692500 passed
## 165   UHM435.20547_S3       13365         1847  13.81967826 passed
## 166  UHM437.20768_S66      148167         1847   1.24656637 passed
## 167  UHM439.20564_S17       96401         1847   1.91595523 passed
## 168    UHM44.3526_S31        5511         1847  33.51478860 failed
## 169  UHM445.20569_S77       90006         1847   2.05208542 passed
## 170  UHM447.20783_S56      168350         1847   1.09711910 passed
## 171  UHM448.20769_S78       28120         1847   6.56827881 passed
## 172    UHM45.3539_S92       35221            0   0.00000000 failed
## 173  UHM454.20770_S90       43632         1847   4.23313165 passed
## 174  UHM455.20785_S80       14926         1847  12.37438028 passed
## 175  UHM458.20786_S92       95135         1847   1.94145162 passed
## 176   UHM459.20787_S9      106958         1847   1.72684605 passed
## 177   UHM461.20771_S7       77653         1847   2.37853013 passed
## 178  UHM467.20772_S19      195315         1847   0.94565190 passed
## 179  UHM470.20533_S25        9207         1847  20.06082329 failed
## 180  UHM476.20414_S46       20085         1847   9.19591735 passed
## 181  UHM478.20549_S27       27792         1847   6.64579735 passed
## 182  UHM479.20551_S51        2116         1847  87.28733459 failed
## 183   UHM481.20403_S9       15762         1847  11.71805608 passed
## 184  UHM482.20590_S44        2014         1847  91.70804369 failed
## 185  UHM483.20603_S10       90381         1847   2.04357110 passed
## 186  UHM519.20582_S43      220555         1847   0.83743284 passed
## 187  UHM520.20573_S30       68381         1847   2.70104269 passed
## 188 UHM746.21478_S117         927          924  99.67637540 failed
## 189 UHM747.21477_S106         924          924 100.00000000 failed
## 190 UHM748.21467_S170         924          924 100.00000000 failed
## 191 UHM748.21487_S129         928          924  99.56896552 failed
## 192 UHM749.21479_S128         925          924  99.89189189 failed
## 193 UHM759.21466_S159         923          923 100.00000000 failed
## 194 UHM759.21486_S118         938          924  98.50746269 failed
## 195 UHM775.21485_S107         923          923 100.00000000 failed
## 196 UHM776.21482_S161         923          923 100.00000000 failed
## 197 UHM777.21484_S183         925          924  99.89189189 failed
## 198 UHM779.21468_S181         928          924  99.56896552 failed
## 199 UHM779.21488_S140         924          924 100.00000000 failed
## 200 UHM782.21480_S139         924          924 100.00000000 failed
## 201 UHM810.21472_S138         923          923 100.00000000 failed
## 202 UHM811.21471_S127         927          924  99.67637540 failed
## 203 UHM813.21481_S150         924          924 100.00000000 failed
## 204 UHM818.21469_S105         924          924 100.00000000 failed
## 205 UHM818.21489_S151        1459          924  63.33104866 failed
## 206 UHM819.21473_S149         923          923 100.00000000 failed
## 207 UHM820.21470_S116         924          924 100.00000000 failed
## 208 UHM820.21490_S162         929          924  99.46178687 failed
## 209 UHM827.21474_S160         935          924  98.82352941 failed
## 210 UHM829.21476_S182         927          924  99.67637540 failed
## 211 UHM832.21483_S172        1028          924  89.88326848 failed
## 212  UHM836.20385_S78        3026            0   0.00000000 failed
## 213  UHM837.20386_S90        2442            0   0.00000000 failed
## 214   UHM838.20387_S7      727048         1847   0.25404100 passed
## 215  UHM891.20384_S66       70359         1847   2.62510837 passed
## 216  UHM892.20532_S13       11192         1847  16.50285919 passed
## 217   UHM893.20595_S9     1153339         1847   0.16014372 passed
## 218  UHM894.20540_S14      291315         1847   0.63402159 passed
## 219  UHM895.20536_S61        5903         1847  31.28917500 failed
## 220  UHM896.20601_S81       10057            0   0.00000000 failed
## 221  UHM897.20591_S56       13724            0   0.00000000 failed
## 222  UHM898.20394_S91         603            0   0.00000000 failed
## 223  UHM899.20588_S20      815235         1847   0.22656044 passed
## 224   UHM900.20395_S8        6596            0   0.00000000 failed
## 225  UHM901.20542_S38      246778         1847   0.74844597 passed
## 226  UHM902.20584_S67        6341         1847  29.12789781 failed
## 227   UHM903.20587_S8       18503         1847   9.98216505 passed
## 228  UHM904.20567_S53      663477         1847   0.27838192 passed
## 229  UHM905.20598_S45       66557         1847   2.77506498 passed
## 230  UHM906.20565_S29       11360            0   0.00000000 failed
## 231  UHM907.20592_S68      378942         1847   0.48740968 passed
## 232  UHM908.20396_S20        8212            0   0.00000000 failed
## 233  UHM909.20557_S28       58573         1847   3.15333003 passed
## 234  UHM910.20562_S88      346860         1847   0.53249150 passed
## 235  UHM965.20537_S73        7079         1847  26.09125583 failed
## 236  UHM966.20743_S51       28439         1847   6.49460248 passed
## 237  UHM967.20744_S63        4222            0   0.00000000 failed
## 238   UHM968.20571_S6        1895         1847  97.46701847 failed
## 239  UHM969.20745_S75      376490         1847   0.49058408 passed
## 240  UHM971.20746_S87     1276457         1847   0.14469739 passed
## 241  UHM973.20578_S90      107606         1847   1.71644704 passed
## 242  UHM974.20432_S72        1133            0   0.00000000 failed
## 243   UHM975.20747_S4      346189         1847   0.53352360 passed
## 244  UHM977.20748_S16      145629         1847   1.26829134 passed
## 245  UHM978.20749_S28      128603         1847   1.43620289 passed
## 246  UHM979.20750_S40      287549         1847   0.64232531 passed
## 247   UHM980.20731_S2       28474         1847   6.48661937 passed
## 248   UHM981.20539_S2       40192         1847   4.59544188 passed
## 249  UHM982.20740_S15       65852         1847   2.80477434 passed
## 250  UHM983.20556_S16        1885         1847  97.98408488 failed
## 251  UHM984.20751_S52      330025         1847   0.55965457 passed
## 252  UHM985.20752_S64        5950         1847  31.04201681 failed
## 253  UHM988.20753_S76       23740         1847   7.78011794 passed
## 254  UHM989.20754_S88      634827         1847   0.29094541 passed
## 255   UHM991.20755_S5        8304         1847  22.24229287 failed
## 256  UHM993.20741_S27      237465         1847   0.77779883 passed
## 257  UHM996.20610_S94       14720            0   0.00000000 failed
## 258  UHM997.20553_S75       12274            0   0.00000000 failed
## 259  UHM998.20618_S95        5965          924  15.49036044 passed
## 260  UHM999.20617_S83       92690          924   0.99687129 passed

# Filter to keep only the samples that passed
passed_samples <- Perc$Sample[Perc$Result == "passed"]

# Subset the TSE object to include only passed samples
tse_passed <- tse_absolute_filtered[, colnames(tse_absolute_filtered) %in% passed_samples]
dim(tse_passed)

## [1] 8742  177

Abundance-based Core microbiome

Alluvial plot illustrating the distribution of prevalent and core microbiome taxa across animal types, ecoregions, diets, and ecological modes. Flows represent absolute abundance of bacterial classes (colored by taxonomic class) across categorical variables. Dominant lineages such as Bacteroidia, Clostridia, Fusobacteria and Gammaproteobacteria remain prevalent across diverse hosts and habitats.

pps_Abs <- DspikeIn::get_long_format_data(tse_passed)

# calculation for relative abundance needs sum of total reads
# total_reads <- sum(pps_Abs$Abundance)

# Generate an alluvial plot

alluvial_plot_abs <- alluvial_plot(
  data = pps_Abs,
  axes = c("Animal.type", "Ecoregion.III","Diet", "Animal.ecomode"),
  abundance_threshold = 5000,
  fill_variable = "Class",
  silent = TRUE,
  abundance_type = "absolute",
  top_taxa = 10,
  text_size = 3,
  legend_ncol = 1,
  custom_colors = DspikeIn::color_palette$MG_Awesome  # Use the color palette from DspikeIn
)

Data transform & Normalization

Normalization is a crucial preprocessing step in microbiome analysis. It corrects for technical artifacts such as unequal sequencing depth, amplification bias, and compositionality, ensuring that observed differences reflect biological variation rather than data imbalance. The choice of normalization depends on the goal of your analysis, differential abundance, compositional analysis, or quick exploratory visualization. Normalization is a crucial preprocessing step in microbiome analysis. It corrects for technical artifacts such as unequal sequencing depth, amplification bias, and compositionality, ensuring that observed differences reflect biological variation rather than data imbalance.
The choice of normalization depends on the goal of your analysis, differential abundance, compositional analysis, or quick exploratory visualization.

Normalization Method	Corrects For	Best For	Notes / Comments
DESeq	Library size and count variance	Differential abundance testing	Robust, model-based normalization from DESeq2.
TMM (Trimmed Mean of M-values)	Library size and compositional bias	Group comparisons	Widely used in edgeR, handles unequal composition.
CSS (Cumulative Sum Scaling)	Sparse, compositional count data	Microbiome datasets with many zeros	Balances low-abundance and high-abundance taxa.
CLR (Centered Log-Ratio)	Compositional structure	Ordination, diversity, correlation	Produces log-ratio data suitable for distance-based methods.
TC (Total Count)	Library size	Simple exploratory work	Quick but sensitive to outliers.
UQ (Upper Quartile)	Library size and extreme counts	Moderate bias correction	Uses the upper quartile of counts as scaling reference.
Median	Library size	Exploratory visualization	Stable for balanced datasets.
RLE (Relative Log Expression)	Sample-specific bias	Large count matrices	Used in RNA-seq and applicable to microbial data.
TSS (Total Sum Scaling)	Library size	Relative abundance conversion	Similar to TC, transforms counts to proportions.
QN (Quantile Normalization)	Global distribution differences	Multi-omics integration	Forces identical distributions across samples.
Poisson	Count noise model	Statistical modeling	Based on Poisson GLM; often paired with inference.
Rarefying	Unequal sequencing depth	Richness/evenness estimation	Not recommended for statistical tests.

Use DESeq, TMM, or CSS for differential abundance testing.
Use CLR for compositional and ordination analyses.
Use TC, UQ, or Median for exploratory or rapid screening.
Always check scaling factors and library size distributions before interpretation.

# you may need to normalize/transform your data to reduce biases

ps <- tse_passed

# TC Normalization
result_TC <- normalization_set(ps, method = "TC", groups = "Host.species")
normalized_ps_TC <- result_TC$dat.normed
scaling_factors_TC <- result_TC$scaling.factor

# UQ Normalization
# result_UQ <- normalization_set(ps, method = "UQ", groups = "Host.species")
# normalized_ps_UQ <- result_UQ$dat.normed
# scaling_factors_UQ <- result_UQ$scaling.factor
# 
# # Median Normalization
# result_med <- normalization_set(ps, method = "med", groups = "Host.species")
# normalized_ps_med <- result_med$dat.normed
# scaling_factors_med <- result_med$scaling.factor

# DESeq Normalization
# ps_n <- remove_zero_negative_count_samples(ps)
# result_DESeq <- normalization_set(ps_n, method = "DESeq", groups = "Animal.type")
# normalized_ps_DESeq <- result_DESeq$dat.normed
# scaling_factors_DESeq <- result_DESeq$scaling.factor

# Poisson Normalization
# result_Poisson <- normalization_set(ps, method = "Poisson", groups = "Host.genus")
# normalized_ps_Poisson <- result_Poisson$dat.normed
# scaling_factors_Poisson <- result_Poisson$scaling.factor

# Quantile Normalization
# result_QN <- normalization_set(ps, method = "QN")
# normalized_ps_QN <- result_QN$dat.normed
# scaling_factors_QN <- result_QN$scaling.factor

# TMM Normalization
# result_TMM <- normalization_set(ps, method = "TMM", groups = "Animal.type")
# normalized_ps_TMM <- result_TMM$dat.normed
# scaling_factors_TMM <- result_TMM$scaling.factor

# CLR Normalization
# result_clr <- normalization_set(ps, method = "clr")
# normalized_ps_clr <- result_clr$dat.normed
# scaling_factors_clr <- result_clr$scaling.factor

# Rarefying
# result_rar <- normalization_set(ps, method = "rar")
# normalized_ps_rar <- result_rar$dat.normed
# scaling_factors_rar <- result_rar$scaling.factor

# CSS Normalization
# result_css <- normalization_set(ps, method = "css")
# normalized_ps_css <- result_css$dat.normed
# scaling_factors_css <- result_css$scaling.factor
# 
# TSS Normalization
# result_tss <- normalization_set(ps, method = "tss")
# normalized_ps_tss <- result_tss$dat.normed
# scaling_factors_tss <- result_tss$scaling.factor

# RLE Normalization
# result_rle <- normalization_set(ps, method = "rle")
# normalized_ps_rle <- result_rle$dat.normed
# scaling_factors_rle <- result_rle$scaling.factor

Random Forestbased feature selection identifies the most predictive ASVs or OTUs that distinguish samples according to a chosen metadata variable (e.g., host, treatment, or environment). It builds an ensemble of decision trees to evaluate how strongly each taxon contributes to accurate classification, ranking taxa by Mean Decrease in Gini impurity (importance score). This approach helps users pinpoint biologically informative taxa while reducing data dimensionality, noise, and redundancy, enabling focused downstream analyses such as differential abundance, biomarker discovery, or visualization of key community drivers.For methodological details, see Liaw & Wiener (2002), Classification and Regression by randomForest, R News, 2(3):1822. Available at: journal.r-project.org/articles/RN-2002-022/RN-2002-022.pdf

Ridge plot

The ridge plot illustrates abundance distributions of the top 10 families across all samples, showing which taxa exhibit narrow, high-density peaks (consistent abundance) versus broader or multimodal patterns (context-dependent abundance)

normalized_tse_TC<-convert_phyloseq_to_tse(normalized_ps_TC)

rf_tse <- RandomForest_selected(
  normalized_ps_TC,
  response_var = "Host.genus",
  na_vars = c("Habitat", "Ecoregion.III", "Host.genus", "Diet")
)

ridge_tse<- ridge_plot_it(rf_tse, taxrank = "Family", top_n = 10) + scale_fill_manual(values = DspikeIn::color_palette$cool_MG)

ridge_tse

Differential abundance (Single Pairwise)

remove the spike-in sp before further analysis Note: Before performing differential abundance analysis, it is recommended to remove the spike-in species to avoid artificial bias in normalization. If you select the DESeq2 option embedded in DspikeIn, note that it internally handles all normalization and variance stabilization steps required for modeling raw count data. DESeq2 estimates size factors (accounting for library depth differences) and dispersion parameters (accounting for biological variability). Therefore, users should always provide integer, raw counts, not normalized, transformed, or rarefied data, as input.

library(mia)
library(dplyr)
library(SummarizedExperiment)

# ---Remove unwanted taxa by Genus, Family, or Order ---
tse_filtered <- tse_absolute[
  rowData(tse_absolute)$Genus != "Tetragenococcus" &
  rowData(tse_absolute)$Family != "Chloroplast" &
  rowData(tse_absolute)$Order != "Chloroplast", ]

tse_caudate <- tse_filtered[, colData(tse_filtered)$Clade.Order == "Caudate"]
genera_keep <- c("Desmognathus", "Plethodon", "Eurycea")
tse_three_genera <- tse_caudate[, colData(tse_caudate)$Host.genus %in% genera_keep]
tse_blue_ridge <- tse_three_genera[, colData(tse_three_genera)$Ecoregion.III == "Blue Ridge"]
tse_desmog <- tse_blue_ridge[, colData(tse_blue_ridge)$Host.genus == "Desmognathus"]

# --- Differential Abundance with DESeq2 ---
results_DESeq2 <- perform_and_visualize_DA(
  obj = tse_desmog,
  method = "DESeq2",
  group_var = "Host.taxon",
  contrast = c("Desmognathus monticola", "Desmognathus imitator"),
  output_csv_path = "DA_DESeq2.csv",
  target_glom = "Genus",
  significance_level = 0.01)


results_DESeq2$results

##        baseMean         logFC      lfcSE          stat       pvalue
## 1      9.000000  9.800678e-08 0.25631907  3.823624e-07 9.999997e-01
## 2      2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 3      1.357143 -1.066791e-06 0.75281614 -1.417068e-06 9.999989e-01
## 4      2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 5     10.000000  9.288232e-08 0.24375252  3.810517e-07 9.999997e-01
## 6     10.809524 -4.623429e-01 0.28405960 -1.627626e+00 1.036041e-01
## 7      3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 8      1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 9     21.000000  6.099293e-08 0.17177970  3.550648e-07 9.999997e-01
## 10     2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 11     1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 12     8.000000  1.038298e-07 0.27119223  3.828642e-07 9.999997e-01
## 13     8.000000  1.038298e-07 0.27119223  3.828642e-07 9.999997e-01
## 14     1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 15    11.000000  8.834475e-08 0.23295174  3.792406e-07 9.999997e-01
## 16    15.000000  7.442266e-08 0.20119424  3.699045e-07 9.999997e-01
## 17    56.761905  1.958166e-01 0.14679711  1.333927e+00 1.822279e-01
## 18     4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 19    11.857143  9.818528e-01 0.36399419  2.697441e+00 6.987473e-03
## 20     9.000000  9.800678e-08 0.25631907  3.823624e-07 9.999997e-01
## 21     1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 22     1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 23    24.976190  8.668560e-01 0.34431626  2.517616e+00 1.181522e-02
## 24     3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 25     3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 26     3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 27    16.404762  5.297905e-08 0.19516750  2.714543e-07 9.999998e-01
## 28     1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 29     1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 30     1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 31     4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 32    12.000000  8.429795e-08 0.22353439  3.771140e-07 9.999997e-01
## 33     4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 34    39.261905  3.630680e+00 0.53099439  6.837511e+00 8.058110e-12
## 35     3.547619 -6.026639e-01 0.44992678 -1.339471e+00 1.804174e-01
## 36     1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 37     2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 38    13.880952  3.975924e-01 0.24231412  1.640814e+00 1.008360e-01
## 39     2.095238  4.752914e-14 0.53727874  8.846272e-14 1.000000e+00
## 40     5.500000  6.629646e-01 0.43627227  1.519612e+00 1.286085e-01
## 41     1.023810 -1.585451e-07 0.75891251 -2.089110e-07 9.999998e-01
## 42     2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 43     5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 44     1.119048  7.004394e-01 0.70215395  9.975581e-01 3.184937e-01
## 45     1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 46    17.142857  7.224661e-01 0.30090805  2.400953e+00 1.635243e-02
## 47    23.619048  1.400377e+00 0.34771811  4.027335e+00 5.641269e-05
## 48     2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 49     2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 50     1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 51     4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 52     5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 53     4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 54     1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 55     1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 56     3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 57     1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 58     3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 59     3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 60     2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 61     1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 62     4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 63     2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 64     2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 65     1.119048  7.004394e-01 0.70215395  9.975581e-01 3.184937e-01
## 66     4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 67     1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 68     5.309524  4.059925e-01 0.33616143  1.207731e+00 2.271509e-01
## 69     4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 70     3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 71     2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 72     5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 73     1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 74    10.000000  9.288232e-08 0.24375252  3.810517e-07 9.999997e-01
## 75    13.000000  8.066982e-08 0.21523219  3.748037e-07 9.999997e-01
## 76     3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 77   289.785714  2.095483e+00 0.93426063  2.242932e+00 2.490117e-02
## 78     2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 79     7.000000  1.104785e-07 0.28918101  3.820394e-07 9.999997e-01
## 80     2.023810  1.051443e-07 0.53603346  1.961524e-07 9.999998e-01
## 81     1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 82    57.261905 -2.426056e-01 0.13201486 -1.837714e+00 6.610453e-02
## 83   123.523810  4.279776e+00 0.55913981  7.654215e+00 1.944970e-14
## 84    10.476190  8.961643e-01 0.35525171  2.522618e+00 1.164849e-02
## 85     2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 86     8.523810 -3.846636e-01 0.28720265 -1.339346e+00 1.804582e-01
## 87     2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 88     1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 89     8.000000  1.038298e-07 0.27119223  3.828642e-07 9.999997e-01
## 90    16.476190  6.864082e-01 0.38990578  1.760446e+00 7.833217e-02
## 91    44.928571  7.548876e-01 0.22492151  3.356227e+00 7.901371e-04
## 92    15.119048 -3.646632e-01 0.38199948 -9.546169e-01 3.397715e-01
## 93     4.571429 -7.369649e-01 0.41573062 -1.772698e+00 7.627875e-02
## 94     2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 95     9.285714  9.285915e-08 0.26173634  3.547813e-07 9.999997e-01
## 96     3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 97     3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 98     1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 99    42.166667 -5.479316e-01 0.20781339 -2.636652e+00 8.372873e-03
## 100    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 101    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 102   12.023810 -1.329674e-02 0.22313721 -5.958995e-02 9.524822e-01
## 103    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 104    5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 105    5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 106    7.000000  1.104785e-07 0.28918101  3.820394e-07 9.999997e-01
## 107    7.238095 -2.125444e-01 0.28667600 -7.414100e-01 4.584449e-01
## 108    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 109    8.000000  1.038298e-07 0.27119223  3.828642e-07 9.999997e-01
## 110   18.285714  6.616396e-08 0.18706336  3.536981e-07 9.999997e-01
## 111   92.785714 -5.424302e-02 0.09194237 -5.899677e-01 5.552123e-01
## 112    6.000000  1.180720e-07 0.31154856  3.789842e-07 9.999997e-01
## 113    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 114    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 115    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 116    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 117    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 118    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 119    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 120    5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 121    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 122    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 123    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 124    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 125    7.000000  1.104785e-07 0.28918101  3.820394e-07 9.999997e-01
## 126    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 127    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 128    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 129    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 130    2.023810  9.168515e-08 0.53603231  1.710441e-07 9.999999e-01
## 131    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 132   41.809524  3.673709e+00 0.61909196  5.934028e+00 2.955915e-09
## 133   12.023810  8.291207e-08 0.22353300  3.709165e-07 9.999997e-01
## 134    5.071429  1.061167e-07 0.34063424  3.115267e-07 9.999998e-01
## 135    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 136    8.000000  1.038298e-07 0.27119223  3.828642e-07 9.999997e-01
## 137    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 138    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 139    6.000000  1.180720e-07 0.31154856  3.789842e-07 9.999997e-01
## 140    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 141    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 142    6.000000  1.180720e-07 0.31154856  3.789842e-07 9.999997e-01
## 143    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 144    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 145    4.333333  5.235620e-01 0.37613254  1.391962e+00 1.639340e-01
## 146   10.000000  6.374300e-01 0.31019799  2.054913e+00 3.988737e-02
## 147    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 148    2.571429  1.169925e+00 0.55078017  2.124123e+00 3.365989e-02
## 149    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 150    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 151    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 152    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 153    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 154    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 155    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 156    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 157    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 158    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 159    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 160    9.000000  9.800678e-08 0.25631907  3.823624e-07 9.999997e-01
## 161    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 162    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 163    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 164    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 165    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 166    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 167    3.261905  4.109917e-08 0.44913022  9.150836e-08 9.999999e-01
## 168    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 169    7.071429 -6.711406e-02 0.28689158 -2.339353e-01 8.150352e-01
## 170    4.380952 -5.305143e-01 0.37017223 -1.433155e+00 1.518135e-01
## 171    1.047619 -2.000837e-07 0.75955890 -2.634210e-07 9.999998e-01
## 172    6.000000  1.180720e-07 0.31154856  3.789842e-07 9.999997e-01
## 173    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 174    3.190476 -3.015274e-07 0.68407553 -4.407809e-07 9.999996e-01
## 175   11.261905  1.966674e+00 0.73265235  2.684321e+00 7.267734e-03
## 176    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 177   10.000000  9.288232e-08 0.24375252  3.810517e-07 9.999997e-01
## 178    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 179   31.309524  6.079477e+00 0.86731515  7.009536e+00 2.391097e-12
## 180    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 181    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 182    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 183    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 184    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 185    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 186    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 187    4.833333  3.076815e+00 0.64315630  4.783930e+00 1.719008e-06
## 188    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 189    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 190    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 191    3.333333 -2.174825e-08 0.44825252 -4.851785e-08 1.000000e+00
## 192    7.642857 -7.842706e-01 0.40627937 -1.930373e+00 5.356068e-02
## 193    3.404762  6.694691e-08 0.47686423  1.403899e-07 9.999999e-01
## 194    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 195   10.571429 -3.410367e-01 0.26036940 -1.309819e+00 1.902572e-01
## 196    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 197    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 198    6.000000  1.180720e-07 0.31154856  3.789842e-07 9.999997e-01
## 199    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 200    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 201    5.214286  4.287320e-08 0.50577755  8.476691e-08 9.999999e-01
## 202   54.738095 -2.655028e+00 0.64109196 -4.141416e+00 3.451690e-05
## 203    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 204    1.476190  1.807354e+00 0.74242143  2.434404e+00 1.491632e-02
## 205    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 206    1.880952  2.491852e+00 0.78995663  3.154416e+00 1.608199e-03
## 207   14.000000  7.739627e-08 0.20783888  3.723859e-07 9.999997e-01
## 208    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 209    1.642857 -1.999995e+00 0.79011012 -2.531287e+00 1.136450e-02
## 210    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 211    6.000000  1.180720e-07 0.31154856  3.789842e-07 9.999997e-01
## 212    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 213    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 214    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 215    1.214286 -8.915171e-07 0.75517729 -1.180540e-06 9.999991e-01
## 216    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 217    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 218    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 219    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 220    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 221    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 222    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 223   19.523810 -4.654397e-01 0.61459794 -7.573076e-01 4.488656e-01
## 224    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 225    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 226    9.000000  9.800678e-08 0.25631907  3.823624e-07 9.999997e-01
## 227   23.619048 -1.976197e-01 0.21423432 -9.224466e-01 3.562957e-01
## 228    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 229    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 230    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 231    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 232    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 233    7.000000  1.104785e-07 0.28918101  3.820394e-07 9.999997e-01
## 234    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 235    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 236    1.071429 -3.594550e-07 0.76014691 -4.728757e-07 9.999996e-01
## 237    6.238095  7.038938e-02 0.44806983  1.570947e-01 8.751702e-01
## 238    7.000000  1.104785e-07 0.28918101  3.820394e-07 9.999997e-01
## 239    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 240    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 241    5.047619  7.038945e-02 0.33688817  2.089401e-01 8.344950e-01
## 242    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 243    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 244    3.452381  8.413023e-01 0.44943107  1.871927e+00 6.121667e-02
## 245    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 246   22.000000  5.928805e-08 0.16808263  3.527316e-07 9.999997e-01
## 247    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 248    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 249   24.785714  2.288188e-01 0.17474106  1.309473e+00 1.903741e-01
## 250    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 251    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 252    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 253    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 254    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 255    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 256    1.190476 -3.542875e-07 0.78478742 -4.514440e-07 9.999996e-01
## 257    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 258    6.476190  5.025004e-01 0.32234720  1.558879e+00 1.190249e-01
## 259    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 260    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 261    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 262    6.000000  1.180720e-07 0.31154856  3.789842e-07 9.999997e-01
## 263    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 264    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 265    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 266    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 267    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 268    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 269    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 270    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 271    8.000000  1.038298e-07 0.27119223  3.828642e-07 9.999997e-01
## 272    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 273   28.214286  7.441612e-01 0.24154540  3.080834e+00 2.064219e-03
## 274    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 275    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 276    6.642857 -8.119271e-01 0.40319144 -2.013751e+00 4.403570e-02
## 277    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 278   27.857143  1.542258e+00 0.26923273  5.728345e+00 1.014149e-08
## 279    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 280    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 281    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 282   17.714286 -4.374051e-01 0.25578074 -1.710078e+00 8.725139e-02
## 283    8.000000  1.038298e-07 0.27119223  3.828642e-07 9.999997e-01
## 284    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 285   16.000000  7.170465e-08 0.19517417  3.673880e-07 9.999997e-01
## 286    1.714286  2.247926e+00 0.77317723  2.907388e+00 3.644609e-03
## 287    6.119048  5.889379e-02 0.30904900  1.905646e-01 8.488668e-01
## 288    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 289   12.000000  8.429795e-08 0.22353439  3.771140e-07 9.999997e-01
## 290    5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 291    5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 292    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 293    9.000000  9.800678e-08 0.25631907  3.823624e-07 9.999997e-01
## 294    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 295    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 296    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 297    2.119048 -3.536364e-01 0.51732889 -6.835814e-01 4.942396e-01
## 298    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 299    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 300    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 301   33.976190  7.750635e-08 0.19184266  4.040100e-07 9.999997e-01
## 302    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 303    9.000000  9.800678e-08 0.25631907  3.823624e-07 9.999997e-01
## 304   13.428571 -1.849176e-01 0.21778616 -8.490787e-01 3.958375e-01
## 305    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 306    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 307   10.000000  9.288232e-08 0.24375252  3.810517e-07 9.999997e-01
## 308   16.000000  7.170465e-08 0.19517417  3.673880e-07 9.999997e-01
## 309    2.023810  1.051443e-07 0.53603346  1.961524e-07 9.999998e-01
## 310    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 311   18.000000  6.691159e-08 0.18465683  3.623564e-07 9.999997e-01
## 312    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 313    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 314    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 315    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 316    5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 317    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 318    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 319    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 320    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 321    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 322    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 323    3.523810 -5.274572e-07 0.60252102 -8.754170e-07 9.999993e-01
## 324    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 325   14.904762  2.370393e-01 0.21851837  1.084757e+00 2.780294e-01
## 326    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 327    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 328    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 329    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 330   26.976190 -1.993087e-01 0.39677064 -5.023273e-01 6.154373e-01
## 331    3.952381 -1.311200e+00 0.53036352 -2.472266e+00 1.342594e-02
## 332    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 333    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 334    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 335    5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 336   41.690476  7.995031e-01 0.19705169  4.057327e+00 4.963756e-05
## 337    5.476190  1.039777e-07 0.37337055  2.784840e-07 9.999998e-01
## 338   14.000000  7.739627e-08 0.20783888  3.723859e-07 9.999997e-01
## 339    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 340    8.000000  1.038298e-07 0.27119223  3.828642e-07 9.999997e-01
## 341    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 342    2.714286  1.523562e+00 0.58284897  2.613990e+00 8.949154e-03
## 343    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 344    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 345    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 346    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 347    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 348    2.190476 -3.109548e-08 0.54509012 -5.704649e-08 1.000000e+00
## 349    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 350    7.000000  1.104785e-07 0.28918101  3.820394e-07 9.999997e-01
## 351    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 352    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 353    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 354    6.000000  1.180720e-07 0.31154856  3.789842e-07 9.999997e-01
## 355    9.000000  9.800678e-08 0.25631907  3.823624e-07 9.999997e-01
## 356    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 357    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 358    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 359    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 360   20.404762  8.955308e-01 0.33963099  2.636776e+00 8.369810e-03
## 361    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 362    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 363    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 364    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 365   14.000000  7.739627e-08 0.20783888  3.723859e-07 9.999997e-01
## 366    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 367   12.214286 -5.994617e-01 0.30785827 -1.947200e+00 5.151075e-02
## 368    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 369    3.023810  1.292256e-07 0.43776072  2.951969e-07 9.999998e-01
## 370    2.476190 -7.083558e-08 0.60222887 -1.176224e-07 9.999999e-01
## 371   16.785714  3.219282e-01 0.19844704  1.622237e+00 1.047526e-01
## 372    6.214286  7.004397e-01 0.40196310  1.742547e+00 8.141272e-02
## 373    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 374    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 375   11.071429  8.524463e-08 0.23309617  3.657058e-07 9.999997e-01
## 376    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 377    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 378    5.285714  3.785117e-01 0.33476840  1.130667e+00 2.581951e-01
## 379    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 380    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 381    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 382    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 383    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 384    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 385    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 386   90.261905 -3.089723e-01 0.17801913 -1.735613e+00 8.263231e-02
## 387    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 388    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 389    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 390    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 391    5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 392    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 393    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 394   21.785714  5.506111e-02 0.17453840  3.154670e-01 7.524071e-01
## 395    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 396    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 397    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 398    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 399    3.309524  6.244909e-01 0.42931395  1.454625e+00 1.457730e-01
## 400    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 401    4.142857  8.746294e-02 0.37568080  2.328118e-01 8.159075e-01
## 402    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 403    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 404    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 405    9.690476  1.584962e+00 0.51555084  3.074308e+00 2.109913e-03
## 406    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 407  184.714286  1.809477e+00 0.78496197  2.305178e+00 2.115658e-02
## 408    5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 409    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 410    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 411   18.190476  9.744181e-08 0.24281600  4.012990e-07 9.999997e-01
## 412    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 413   79.928571 -1.929264e+00 0.65963643 -2.924738e+00 3.447466e-03
## 414    7.000000  1.104785e-07 0.28918101  3.820394e-07 9.999997e-01
## 415    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 416    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 417    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 418    8.000000  1.038298e-07 0.27119223  3.828642e-07 9.999997e-01
## 419    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 420    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 421    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 422    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 423    5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 424    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 425    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 426    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 427    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 428    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 429    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 430    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 431    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 432    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 433    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 434    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 435    6.000000  1.180720e-07 0.31154856  3.789842e-07 9.999997e-01
## 436    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 437    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 438    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 439   16.571429  1.197752e-07 0.44679322  2.680775e-07 9.999998e-01
## 440   18.428571 -2.047302e+00 0.69180060 -2.959382e+00 3.082571e-03
## 441    6.595238  9.831783e-09 0.31120125  3.159301e-08 1.000000e+00
## 442    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 443    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 444    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 445    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 446    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 447    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 448    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 449   13.000000  8.066982e-08 0.21523219  3.748037e-07 9.999997e-01
## 450    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 451    5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 452    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 453    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 454    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 455    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 456    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 457    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 458    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 459    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 460    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 461    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 462    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 463    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 464    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 465    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 466    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 467    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 468    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 469    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 470    5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 471    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 472    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 473    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 474    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 475    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 476    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 477    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 478    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 479    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 480    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 481    5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 482   10.000000  9.288232e-08 0.24375252  3.810517e-07 9.999997e-01
## 483    8.000000  1.038298e-07 0.27119223  3.828642e-07 9.999997e-01
## 484    1.261905 -6.462306e-07 0.78607290 -8.221001e-07 9.999993e-01
## 485    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 486    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 487    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 488    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 489    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 490    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 491    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 492    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 493    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 494    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 495    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 496    1.071429 -2.725581e-07 0.76104209 -3.581380e-07 9.999997e-01
## 497   24.547619  6.490929e-01 0.24181767  2.684225e+00 7.269825e-03
## 498    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 499   18.619048 -5.305144e-01 0.27273903 -1.945136e+00 5.175866e-02
## 500    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 501    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 502    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 503   14.809524  7.752938e-01 0.31667031  2.448268e+00 1.435448e-02
## 504    3.047619  1.154773e-01 0.43033729  2.683415e-01 7.884365e-01
## 505   15.023810  7.411871e-08 0.20119386  3.683945e-07 9.999997e-01
## 506    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 507   11.000000  8.834475e-08 0.23295174  3.792406e-07 9.999997e-01
## 508    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 509   15.000000  7.442266e-08 0.20119424  3.699045e-07 9.999997e-01
## 510   16.000000  7.170465e-08 0.19517417  3.673880e-07 9.999997e-01
## 511   58.619048  2.151376e-01 0.14374648  1.496646e+00 1.344853e-01
## 512    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 513    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 514    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 515    7.000000  1.104785e-07 0.28918101  3.820394e-07 9.999997e-01
## 516    6.000000  1.180720e-07 0.31154856  3.789842e-07 9.999997e-01
## 517    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 518    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 519    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 520    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 521    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 522    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 523    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 524   10.000000  9.288232e-08 0.24375252  3.810517e-07 9.999997e-01
## 525    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 526    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 527    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 528    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 529    3.238095 -4.545654e-01 0.42904019 -1.059494e+00 2.893750e-01
## 530  178.642857  5.527386e-01 0.14195904  3.893648e+00 9.874772e-05
## 531    2.095238  3.219281e-01 0.51421984  6.260516e-01 5.312811e-01
## 532    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 533    5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 534    3.047619  1.154773e-01 0.43033729  2.683415e-01 7.884365e-01
## 535    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 536   12.333333 -9.060245e-02 0.22294416 -4.063908e-01 6.844555e-01
## 537   15.023810  1.209453e+00 0.36818751  3.284885e+00 1.020242e-03
## 538    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 539  109.809524 -9.109930e-01 0.61333263 -1.485316e+00 1.374600e-01
## 540 2033.833333 -1.261662e+00 0.86122896 -1.464955e+00 1.429332e-01
## 541    9.285714 -1.679445e-01 0.25480346 -6.591139e-01 5.098227e-01
## 542    4.214286  2.028803e-08 0.37936474  5.347896e-08 1.000000e+00
## 543    5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 544    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 545    1.047619 -2.000837e-07 0.75955890 -2.634210e-07 9.999998e-01
## 546    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 547    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 548    8.000000  1.038298e-07 0.27119223  3.828642e-07 9.999997e-01
## 549   26.547619 -6.627316e-02 0.15647025 -4.235512e-01 6.718932e-01
## 550    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 551    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 552    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 553    5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 554    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 555    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 556    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 557    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 558    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 559    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 560    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 561    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 562    2.761905 -9.593567e-01 0.58332048 -1.644648e+00 1.000425e-01
## 563    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 564    3.142857  9.853515e-08 0.44045283  2.237133e-07 9.999998e-01
## 565    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 566    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 567    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 568    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 569    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 570    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 571    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 572    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 573    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 574    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 575    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 576    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 577   56.071429  5.115227e+00 0.65520052  7.807117e+00 5.851095e-15
## 578    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 579    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 580    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 581    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 582    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 583    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 584   11.023810  8.670545e-08 0.23294962  3.722069e-07 9.999997e-01
## 585    7.690476  1.415036e+00 0.58377010  2.423962e+00 1.535222e-02
## 586    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 587    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 588    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 589    2.333333 -1.198686e-07 0.55395378 -2.163873e-07 9.999998e-01
## 590    5.523810 -5.742356e-01 0.36511832 -1.572739e+00 1.157794e-01
## 591    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 592    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 593    7.190476  1.026353e-07 0.29212123  3.513450e-07 9.999997e-01
## 594    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 595    1.380952 -1.473928e+00 0.74888340 -1.968167e+00 4.904878e-02
## 596    3.476190  8.744691e-01 0.45309897  1.929974e+00 5.361009e-02
## 597    9.404762 -8.634974e-01 0.33100577 -2.608708e+00 9.088473e-03
## 598   14.880952  3.423923e-01 0.22883366  1.496250e+00 1.345886e-01
## 599    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 600    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 601    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 602   92.595238  1.455095e+00 0.33336854  4.364823e+00 1.272257e-05
## 603    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 604    3.119048 -2.451122e-01 0.42691111 -5.741527e-01 5.658645e-01
## 605    3.428571 -1.169128e-07 0.43648640 -2.678498e-07 9.999998e-01
## 606    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 607    3.547619 -1.370210e-07 0.43616854 -3.141469e-07 9.999997e-01
## 608    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 609    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 610    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 611    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 612    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 613    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 614    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 615    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 616    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 617    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 618    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 619    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 620    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 621 2987.071429 -6.385437e+00 1.25609523 -5.083561e+00 3.704227e-07
## 622    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 623    8.428571 -8.682093e-01 0.38912493 -2.231184e+00 2.566894e-02
## 624    5.571429  1.051281e+00 0.63619524  1.652450e+00 9.844282e-02
## 625    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 626    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 627    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 628    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 629    6.000000  1.180720e-07 0.31154856  3.789842e-07 9.999997e-01
## 630    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 631  193.571429 -6.618573e-01 0.51738510 -1.279235e+00 2.008142e-01
## 632  151.119048 -4.136229e-02 0.22292570 -1.855429e-01 8.528032e-01
## 633    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 634    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 635   82.380952 -4.932838e-01 0.25340082 -1.946654e+00 5.157620e-02
## 636    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 637    7.000000  1.104785e-07 0.28918101  3.820394e-07 9.999997e-01
## 638    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 639  271.738095 -3.917528e+00 0.94505398 -4.145296e+00 3.393756e-05
## 640   57.404762 -4.703350e-02 0.10986714 -4.280943e-01 6.685825e-01
## 641    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 642    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 643    5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 644    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 645    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 646    1.619048 -1.044188e-06 0.82336909 -1.268190e-06 9.999990e-01
## 647    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 648    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 649    9.619048  5.849629e-01 0.46200245  1.266147e+00 2.054605e-01
## 650   21.476190  6.229988e-08 0.17796997  3.500584e-07 9.999997e-01
## 651   15.714286 -7.108252e-02 0.40677458 -1.747467e-01 8.612787e-01
## 652  221.119048 -1.188808e-01 0.50290804 -2.363868e-01 8.131325e-01
## 653    5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 654    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 655    9.000000  9.800678e-08 0.25631907  3.823624e-07 9.999997e-01
## 656    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 657  130.952381 -9.912289e-01 1.14558930 -8.652567e-01 3.868979e-01
## 658  163.428571 -3.151660e+00 0.83813103 -3.760343e+00 1.696807e-04
## 659    6.000000  1.180720e-07 0.31154856  3.789842e-07 9.999997e-01
## 660    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 661    6.000000  1.180720e-07 0.31154856  3.789842e-07 9.999997e-01
## 662 1105.238095  1.787661e-01 0.58307743  3.065906e-01 7.591550e-01
## 663    4.714286 -4.855767e-07 0.51930931 -9.350433e-07 9.999993e-01
## 664  837.523810  8.841921e-01 0.54991875  1.607859e+00 1.078660e-01
## 665  553.238095  6.246340e-01 0.45824073  1.363113e+00 1.728468e-01
## 666    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 667 1670.166667  1.834971e+00 0.70672794  2.596432e+00 9.419758e-03
## 668  674.642857  1.265540e+00 0.83798397  1.510219e+00 1.309875e-01
## 669   99.428571 -1.342672e-01 0.58150523 -2.308959e-01 8.173957e-01
## 670    7.857143 -6.520762e-01 0.33201810 -1.963978e+00 4.953265e-02
## 671    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 672   51.452381  2.515441e-02 0.57437793  4.379418e-02 9.650685e-01
## 673   56.928571  6.057442e-01 0.26227610  2.309567e+00 2.091214e-02
## 674    2.404762 -9.593566e-01 0.54876264 -1.748218e+00 8.042631e-02
## 675   75.976190 -6.190991e-01 0.49139601 -1.259878e+00 2.077133e-01
## 676    5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 677  238.785714  2.713719e+00 0.84793800  3.200374e+00 1.372494e-03
## 678  651.928571 -1.622606e+00 0.86417596 -1.877634e+00 6.043121e-02
## 679    6.071429 -2.163972e+00 0.61435181 -3.522366e+00 4.277128e-04
## 680   19.785714 -7.186518e-01 0.52161688 -1.377739e+00 1.682840e-01
## 681    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 682  327.785714  3.328273e-01 0.55794824  5.965200e-01 5.508279e-01
## 683   45.833333 -6.123756e-02 0.19066955 -3.211712e-01 7.480807e-01
## 684    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 685    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 686    1.523810  1.906890e+00 0.74935885  2.544695e+00 1.093733e-02
## 687    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 688    1.047619 -1.127796e-07 0.75989002 -1.484157e-07 9.999999e-01
## 689   18.595238 -5.399766e-07 0.43047783 -1.254366e-06 9.999990e-01
## 690   17.547619  8.806100e-08 0.24339812  3.617982e-07 9.999997e-01
## 691   95.309524  5.016194e-01 0.86458167  5.801874e-01 5.617882e-01
## 692  514.928571 -8.694313e-01 0.69078714 -1.258610e+00 2.081714e-01
## 693    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 694    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 695   66.190476 -3.546648e-01 0.15661751 -2.264528e+00 2.354164e-02
## 696    8.000000  1.038298e-07 0.27119223  3.828642e-07 9.999997e-01
## 697    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 698    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 699    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 700    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 701    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 702    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 703    2.809524 -1.530512e+00 0.61176392 -2.501802e+00 1.235630e-02
## 704    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 705    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 706    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 707    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 708    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 709    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 710    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 711    4.023810 -3.952820e-02 0.37763867 -1.046720e-01 9.166361e-01
## 712    3.214286 -1.938596e+00 0.65388699 -2.964726e+00 3.029527e-03
## 713    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 714    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 715    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 716    7.095238 -8.889678e-01 0.40152889 -2.213957e+00 2.683171e-02
## 717  576.190476  3.760845e+00 0.65488946  5.742718e+00 9.316901e-09
## 718    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 719    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 720    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 721   18.785714 -8.097060e-01 0.46186466 -1.753124e+00 7.958071e-02
## 722   33.880952 -1.662518e+00 0.58489734 -2.842411e+00 4.477380e-03
## 723    9.095238  9.210198e-08 0.25667793  3.588231e-07 9.999997e-01
## 724    3.404762 -7.045433e-01 0.45071335 -1.563174e+00 1.180117e-01
## 725    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 726    2.095238  3.219281e-01 0.51421984  6.260516e-01 5.312811e-01
## 727    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 728    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 729    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 730    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 731    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 732    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 733   10.333333 -2.085865e-01 0.24489443 -8.517404e-01 3.943582e-01
## 734    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 735    4.333333 -2.410326e-07 0.43452529 -5.547033e-07 9.999996e-01
## 736   31.357143 -2.093105e+00 0.67716953 -3.090962e+00 1.995092e-03
## 737   54.785714 -1.121375e+00 0.58730107 -1.909370e+00 5.621438e-02
## 738    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 739    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 740 1316.738095  2.636997e-01 0.69934967  3.770642e-01 7.061259e-01
## 741    8.571429 -4.150372e-01 0.29220745 -1.420351e+00 1.555054e-01
## 742    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 743    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 744    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 745    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 746    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 747    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 748    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 749    5.047619  1.168349e-07 0.34050335  3.431242e-07 9.999997e-01
## 750    5.023810  1.191098e-07 0.34039621  3.499153e-07 9.999997e-01
## 751    5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 752    8.166667  9.837610e-08 0.27306498  3.602663e-07 9.999997e-01
## 753  394.904762  3.505180e-01 0.42638067  8.220776e-01 4.110327e-01
## 754    7.000000  1.104785e-07 0.28918101  3.820394e-07 9.999997e-01
## 755 1998.857143  4.676185e-01 1.00803520  4.638910e-01 6.427258e-01
## 756 2726.833333 -2.609870e+00 1.23809972 -2.107964e+00 3.503411e-02
## 757    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 758    6.000000  1.180720e-07 0.31154856  3.789842e-07 9.999997e-01
## 759    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 760   16.833333  3.076266e-02 0.33307459  9.235967e-02 9.264123e-01
## 761    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 762   50.357143 -4.855454e-02 0.34803657 -1.395099e-01 8.890472e-01
## 763  153.833333 -5.933346e+00 0.93630174 -6.337003e+00 2.342782e-10
## 764    6.000000  1.180720e-07 0.31154856  3.789842e-07 9.999997e-01
## 765   12.380952 -8.568743e-01 0.38057004 -2.251555e+00 2.435042e-02
## 766    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 767    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 768    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 769  644.785714 -2.202040e-01 0.74702514 -2.947746e-01 7.681661e-01
## 770    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 771    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 772    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 773    2.976190 -1.712715e+00 0.63113476 -2.713707e+00 6.653493e-03
## 774    1.309524 -5.178321e-07 0.81345272 -6.365853e-07 9.999995e-01
## 775    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 776    2.571429 -1.222391e+00 0.57777242 -2.115696e+00 3.437071e-02
## 777    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 778    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 779    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 780    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 781    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 782   33.595238 -1.148521e+00 0.41053581 -2.797616e+00 5.148133e-03
## 783    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 784    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 785    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 786    1.404762 -1.530511e+00 0.75319795 -2.032017e+00 4.215190e-02
## 787    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 788    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 789    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 790    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 791    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 792    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 793    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 794   30.619048 -1.378687e-02 0.18890454 -7.298327e-02 9.418194e-01
## 795  147.452381  1.204936e+00 0.46213767  2.607310e+00 9.125662e-03
## 796    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 797    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 798    8.809524  5.545889e-01 0.29089497  1.906492e+00 5.658640e-02
## 799    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 800    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 801    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 802    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 803   14.000000  7.739627e-08 0.20783888  3.723859e-07 9.999997e-01
## 804    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 805    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 806    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 807    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 808    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 809    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 810    1.380952 -7.646135e-07 0.81522985 -9.379116e-07 9.999993e-01
## 811    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 812    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 813    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 814    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 815    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 816    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 817    6.785714 -4.150364e-01 0.43861076 -9.462521e-01 3.440200e-01
## 818  305.523810 -1.936819e-01 0.67852734 -2.854444e-01 7.753037e-01
## 819   90.214286 -1.520030e-01 0.25220329 -6.027002e-01 5.467081e-01
## 820    2.619048  8.073545e-01 0.53977172  1.495733e+00 1.347232e-01
## 821    8.214286  1.898247e-01 0.26727361  7.102260e-01 4.775640e-01
## 822    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 823    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 824    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 825    4.071429  1.141068e-07 0.38012813  3.001800e-07 9.999998e-01
## 826    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 827    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 828    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 829    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 830    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 831    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 832   50.023810 -2.216534e-01 0.19150406 -1.157435e+00 2.470949e-01
## 833    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 834  264.595238  1.652815e+00 0.70246660  2.352873e+00 1.862901e-02
## 835    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 836    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 837  759.238095  2.952177e+00 0.74505994  3.962335e+00 7.422032e-05
## 838    3.857143 -1.323118e-08 0.53685080 -2.464592e-08 1.000000e+00
## 839    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 840    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 841   16.619048 -5.156412e-01 0.24251446 -2.126229e+00 3.348422e-02
## 842   59.928571  2.154682e+00 0.54401559  3.960698e+00 7.473091e-05
## 843    4.000000  1.357238e-07 0.37974528  3.574074e-07 9.999997e-01
## 844    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 845    6.523810  5.443206e-01 0.32780062  1.660523e+00 9.680919e-02
## 846  449.880952  3.589015e+00 0.64572813  5.558090e+00 2.727435e-08
## 847    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 848    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 849    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 850    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 851    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 852    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 853 2171.000000 -2.831305e-01 0.70457555 -4.018454e-01 6.877978e-01
## 854    9.809524  5.025004e-01 0.28678585  1.752180e+00 7.974286e-02
## 855    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 856    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 857    1.952381 -2.444778e+00 0.82420785 -2.966216e+00 3.014889e-03
## 858    5.000000  1.266256e-07 0.34042060  3.719681e-07 9.999997e-01
## 859    7.000000  1.104785e-07 0.28918101  3.820394e-07 9.999997e-01
## 860    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 861    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 862    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 863    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 864    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 865    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 866    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 867    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 868    1.833333 -2.289501e+00 0.81252059 -2.817776e+00 4.835759e-03
## 869    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 870    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 871    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 872    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 873    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 874    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 875    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 876    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 877    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 878    1.238095 -6.097646e-07 0.78262977 -7.791227e-07 9.999994e-01
## 879    8.476190  1.643856e+00 0.43937811  3.741325e+00 1.830529e-04
## 880    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 881   17.738095 -3.325752e-01 0.25115375 -1.324190e+00 1.854401e-01
## 882    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 883    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 884    3.809524 -1.175848e+00 0.51284308 -2.292803e+00 2.185933e-02
## 885    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 886 4663.142857  2.682200e+00 0.79947632  3.354946e+00 7.938060e-04
## 887    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 888  504.738095  7.283374e-02 0.19270942  3.779459e-01 7.054708e-01
## 889  189.357143 -3.464532e-01 1.11283742 -3.113242e-01 7.555542e-01
## 890    4.023810  1.241637e-07 0.37970153  3.270035e-07 9.999997e-01
## 891    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 892    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 893    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 894    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 895    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 896   36.238095  1.051453e-07 0.23102846  4.551186e-07 9.999996e-01
## 897    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 898    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 899    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 900    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 901    7.047619  1.029291e-07 0.28922569  3.558783e-07 9.999997e-01
## 902    9.404762  9.425146e-01 0.33710408  2.795916e+00 5.175290e-03
## 903    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 904   11.047619  8.516365e-08 0.23298543  3.655321e-07 9.999997e-01
## 905    3.000000  1.429881e-07 0.43784445  3.265728e-07 9.999997e-01
## 906    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 907    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 908    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 909    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 910  217.976190 -4.616086e+00 1.01017512 -4.569590e+00 4.886798e-06
## 911    1.166667 -4.873779e-07 0.77217684 -6.311740e-07 9.999995e-01
## 912    3.261905 -4.930395e-01 0.43106652 -1.143767e+00 2.527205e-01
## 913   70.452381  5.911150e-02 0.80252445  7.365695e-02 9.412834e-01
## 914  904.809524 -2.876568e-01 0.51645853 -5.569795e-01 5.775414e-01
## 915 1244.666667 -2.723134e+00 0.98482067 -2.765106e+00 5.690429e-03
## 916 3495.809524 -2.343358e-01 0.73738977 -3.177910e-01 7.506435e-01
## 917 1464.404762 -7.688620e-02 0.84167278 -9.134928e-02 9.272151e-01
## 918    1.714286  2.247926e+00 0.77317723  2.907388e+00 3.644609e-03
## 919    9.761905 -4.803286e-01 0.29510902 -1.627631e+00 1.036031e-01
## 920    4.761905 -9.175369e-01 0.44315196 -2.070479e+00 3.840750e-02
## 921 8241.833333  2.019912e+00 0.67208425  3.005444e+00 2.651932e-03
## 922    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 923   22.071429 -3.018226e-08 0.52312974 -5.769556e-08 1.000000e+00
## 924    3.071429 -1.520028e-01 0.42974112 -3.537079e-01 7.235578e-01
## 925 1521.809524  3.868149e+00 1.11694597  3.463148e+00 5.338948e-04
## 926  102.904762 -1.678058e+00 0.95162351 -1.763363e+00 7.783925e-02
## 927    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 928    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 929    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 930    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 931  343.761905  4.252577e-01 0.20767612  2.047697e+00 4.058971e-02
## 932   11.476190 -9.999990e-01 0.44343445 -2.255122e+00 2.412565e-02
## 933   25.309524  5.470722e-08 0.16030336  3.412731e-07 9.999997e-01
## 934    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 935    9.833333  4.447849e-01 0.28279579  1.572813e+00 1.157621e-01
## 936    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 937    1.047619 -2.895059e-01 0.73181850 -3.955979e-01 6.924017e-01
## 938    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 939    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 940 1332.500000 -5.101195e-02 0.85399526 -5.973329e-02 9.523681e-01
## 941    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 942    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 943    8.309524  9.708649e-08 0.38834496  2.500007e-07 9.999998e-01
## 944   10.071429 -1.349293e-01 0.28696408 -4.701959e-01 6.382151e-01
## 945    3.571429 -9.175368e-01 0.47825011 -1.918529e+00 5.504395e-02
## 946    2.000000  1.353086e-07 0.53621882  2.523383e-07 9.999998e-01
## 947  365.761905 -7.916393e-01 0.93197509 -8.494211e-01 3.956470e-01
## 948    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 949  218.690476  1.691970e-01 0.17613469  9.606116e-01 3.367475e-01
## 950 1009.571429  1.199853e+00 0.99873269  1.201376e+00 2.296055e-01
## 951    2.119048 -3.536364e-01 0.51732889 -6.835814e-01 4.942396e-01
## 952    1.476190 -7.122598e-07 0.84585207 -8.420618e-07 9.999993e-01
## 953   25.071429 -1.176514e-01 0.21314946 -5.519669e-01 5.809710e-01
## 954   17.928571 -1.078001e+00 0.40536328 -2.659347e+00 7.829238e-03
## 955  476.928571  9.207448e-01 0.77347294  1.190403e+00 2.338879e-01
## 956    1.000000  1.223014e-16 0.75942345  1.610450e-16 1.000000e+00
## 957    2.404762 -9.593566e-01 0.54876264 -1.748218e+00 8.042631e-02
## 958    4.214286 -1.530512e+00 0.55722547 -2.746666e+00 6.020436e-03
## 959   17.119048  6.713848e-08 0.18998824  3.533823e-07 9.999997e-01
## 960 8836.547619  4.764254e-01 0.77008025  6.186698e-01 5.361339e-01
##             padj          FDR    Significance                  group     OTU
## 1   1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator    OTU2
## 2   1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator    OTU3
## 3   1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola    OTU4
## 4   1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator    OTU5
## 5   1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator    OTU6
## 6   8.724557e-01 8.724557e-01 Not Significant Desmognathus monticola   OTU10
## 7   1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator   OTU38
## 8   1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator   OTU41
## 9   1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator   OTU42
## 10  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator   OTU45
## 11  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator   OTU48
## 12  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator   OTU61
## 13  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator   OTU68
## 14  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator   OTU75
## 15  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator   OTU79
## 16  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator   OTU83
## 17  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU103
## 18  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU158
## 19  1.424292e-01 1.424292e-01 Not Significant  Desmognathus imitator  OTU162
## 20  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU172
## 21  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU179
## 22  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU180
## 23  1.890435e-01 1.890435e-01 Not Significant  Desmognathus imitator  OTU188
## 24  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU207
## 25  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU208
## 26  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU212
## 27  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU225
## 28  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU237
## 29  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU238
## 30  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU240
## 31  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU241
## 32  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU245
## 33  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU259
## 34  1.933946e-09 1.933946e-09     Significant  Desmognathus imitator  OTU270
## 35  1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola  OTU277
## 36  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU278
## 37  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU282
## 38  8.643085e-01 8.643085e-01 Not Significant  Desmognathus imitator  OTU287
## 39  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU288
## 40  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU295
## 41  1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola  OTU297
## 42  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU298
## 43  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU306
## 44  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU322
## 45  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU323
## 46  2.378536e-01 2.378536e-01 Not Significant  Desmognathus imitator  OTU327
## 47  3.185658e-03 3.185658e-03     Significant  Desmognathus imitator  OTU333
## 48  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU349
## 49  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU351
## 50  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU353
## 51  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU354
## 52  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU355
## 53  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU356
## 54  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU359
## 55  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU363
## 56  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU364
## 57  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU367
## 58  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU371
## 59  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU376
## 60  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU378
## 61  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU385
## 62  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU392
## 63  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU397
## 64  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU424
## 65  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU425
## 66  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU427
## 67  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU429
## 68  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU432
## 69  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU433
## 70  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU435
## 71  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU437
## 72  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU441
## 73  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU446
## 74  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU447
## 75  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU450
## 76  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU451
## 77  3.230422e-01 3.230422e-01 Not Significant  Desmognathus imitator  OTU464
## 78  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU469
## 79  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU470
## 80  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU476
## 81  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU478
## 82  6.475546e-01 6.475546e-01 Not Significant Desmognathus monticola  OTU497
## 83  9.335854e-12 9.335854e-12     Significant  Desmognathus imitator  OTU501
## 84  1.890435e-01 1.890435e-01 Not Significant  Desmognathus imitator  OTU512
## 85  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU527
## 86  1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola  OTU531
## 87  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU539
## 88  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU542
## 89  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU543
## 90  7.353263e-01 7.353263e-01 Not Significant  Desmognathus imitator  OTU549
## 91  2.930976e-02 2.930976e-02 Not Significant  Desmognathus imitator  OTU553
## 92  1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola  OTU576
## 93  7.353263e-01 7.353263e-01 Not Significant Desmognathus monticola  OTU577
## 94  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU580
## 95  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU620
## 96  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU621
## 97  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU623
## 98  1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU625
## 99  1.545761e-01 1.545761e-01 Not Significant Desmognathus monticola  OTU630
## 100 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU635
## 101 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU638
## 102 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola  OTU648
## 103 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU692
## 104 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU698
## 105 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU700
## 106 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU719
## 107 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola  OTU735
## 108 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU741
## 109 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU751
## 110 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU763
## 111 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola  OTU780
## 112 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU790
## 113 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU791
## 114 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU794
## 115 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU795
## 116 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU796
## 117 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU797
## 118 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU800
## 119 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU801
## 120 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU804
## 121 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU805
## 122 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU811
## 123 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU812
## 124 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU813
## 125 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU814
## 126 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU820
## 127 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU824
## 128 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU826
## 129 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU827
## 130 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU828
## 131 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU831
## 132 4.729464e-07 4.729464e-07     Significant  Desmognathus imitator  OTU836
## 133 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU838
## 134 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU841
## 135 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU846
## 136 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU848
## 137 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU854
## 138 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU857
## 139 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU863
## 140 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU865
## 141 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU870
## 142 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU871
## 143 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU873
## 144 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU874
## 145 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU879
## 146 4.669741e-01 4.669741e-01 Not Significant  Desmognathus imitator  OTU883
## 147 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU884
## 148 4.142756e-01 4.142756e-01 Not Significant  Desmognathus imitator  OTU885
## 149 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU889
## 150 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU890
## 151 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU909
## 152 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU912
## 153 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU913
## 154 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU915
## 155 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU916
## 156 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU925
## 157 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU932
## 158 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU933
## 159 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU934
## 160 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU935
## 161 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU939
## 162 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU941
## 163 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU943
## 164 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU944
## 165 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU946
## 166 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU948
## 167 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU954
## 168 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU955
## 169 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola  OTU956
## 170 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola  OTU965
## 171 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola  OTU968
## 172 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU969
## 173 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU972
## 174 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola  OTU976
## 175 1.424292e-01 1.424292e-01 Not Significant  Desmognathus imitator  OTU978
## 176 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU982
## 177 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU984
## 178 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU987
## 179 7.651510e-10 7.651510e-10     Significant  Desmognathus imitator  OTU988
## 180 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU991
## 181 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU993
## 182 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator  OTU998
## 183 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1004
## 184 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1007
## 185 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1010
## 186 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1011
## 187 1.500225e-04 1.500225e-04     Significant  Desmognathus imitator OTU1013
## 188 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1014
## 189 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1016
## 190 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1018
## 191 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU1021
## 192 5.594096e-01 5.594096e-01 Not Significant Desmognathus monticola OTU1022
## 193 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1024
## 194 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1025
## 195 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU1029
## 196 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1051
## 197 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1058
## 198 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1060
## 199 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1068
## 200 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1069
## 201 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1071
## 202 2.209082e-03 2.209082e-03     Significant Desmognathus monticola OTU1083
## 203 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1089
## 204 2.237448e-01 2.237448e-01 Not Significant  Desmognathus imitator OTU1094
## 205 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1095
## 206 5.323692e-02 5.323692e-02 Not Significant  Desmognathus imitator OTU1096
## 207 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1097
## 208 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1099
## 209 1.881020e-01 1.881020e-01 Not Significant Desmognathus monticola OTU1101
## 210 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1102
## 211 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1103
## 212 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1105
## 213 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1106
## 214 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1107
## 215 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU1109
## 216 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1110
## 217 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1113
## 218 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1114
## 219 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1118
## 220 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1120
## 221 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1122
## 222 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1127
## 223 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU1135
## 224 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1138
## 225 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1139
## 226 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1142
## 227 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU1151
## 228 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1159
## 229 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1160
## 230 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1161
## 231 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1162
## 232 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1165
## 233 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1166
## 234 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1167
## 235 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1169
## 236 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU1173
## 237 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1174
## 238 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1175
## 239 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1182
## 240 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1185
## 241 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1190
## 242 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1200
## 243 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1211
## 244 6.058557e-01 6.058557e-01 Not Significant  Desmognathus imitator OTU1213
## 245 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1221
## 246 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1225
## 247 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1226
## 248 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1238
## 249 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1245
## 250 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1266
## 251 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1267
## 252 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1268
## 253 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1272
## 254 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1273
## 255 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1274
## 256 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU1278
## 257 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1279
## 258 9.521993e-01 9.521993e-01 Not Significant  Desmognathus imitator OTU1283
## 259 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1286
## 260 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1289
## 261 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1291
## 262 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1292
## 263 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1296
## 264 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1304
## 265 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1306
## 266 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1315
## 267 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1316
## 268 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1320
## 269 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1325
## 270 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1327
## 271 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1333
## 272 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1341
## 273 6.329740e-02 6.329740e-02 Not Significant  Desmognathus imitator OTU1348
## 274 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1359
## 275 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1378
## 276 4.973444e-01 4.973444e-01 Not Significant Desmognathus monticola OTU1398
## 277 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1399
## 278 1.216979e-06 1.216979e-06     Significant  Desmognathus imitator OTU1400
## 279 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1411
## 280 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1415
## 281 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1423
## 282 7.755680e-01 7.755680e-01 Not Significant Desmognathus monticola OTU1427
## 283 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1434
## 284 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1437
## 285 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1439
## 286 8.971344e-02 8.971344e-02 Not Significant  Desmognathus imitator OTU1455
## 287 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1460
## 288 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1464
## 289 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1467
## 290 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1477
## 291 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1482
## 292 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1483
## 293 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1496
## 294 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1505
## 295 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1506
## 296 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1507
## 297 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU1509
## 298 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1512
## 299 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1514
## 300 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1517
## 301 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1518
## 302 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1522
## 303 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1523
## 304 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU1534
## 305 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1558
## 306 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1561
## 307 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1565
## 308 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1593
## 309 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1594
## 310 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1598
## 311 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1599
## 312 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1601
## 313 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1602
## 314 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1603
## 315 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1604
## 316 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1607
## 317 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1609
## 318 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1612
## 319 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1613
## 320 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1614
## 321 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1615
## 322 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1616
## 323 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU1622
## 324 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1623
## 325 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1629
## 326 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1633
## 327 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1646
## 328 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1648
## 329 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1649
## 330 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU1661
## 331 2.078856e-01 2.078856e-01 Not Significant Desmognathus monticola OTU1670
## 332 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1672
## 333 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1679
## 334 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1680
## 335 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1682
## 336 2.978253e-03 2.978253e-03     Significant  Desmognathus imitator OTU1701
## 337 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1703
## 338 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1713
## 339 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1714
## 340 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1715
## 341 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1726
## 342 1.592843e-01 1.592843e-01 Not Significant  Desmognathus imitator OTU1730
## 343 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1731
## 344 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1736
## 345 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1737
## 346 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1743
## 347 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1775
## 348 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU1776
## 349 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1777
## 350 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1785
## 351 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1791
## 352 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1795
## 353 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1817
## 354 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1818
## 355 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1821
## 356 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1846
## 357 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1848
## 358 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1851
## 359 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1855
## 360 1.545761e-01 1.545761e-01 Not Significant  Desmognathus imitator OTU1875
## 361 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1880
## 362 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1888
## 363 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1890
## 364 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1891
## 365 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1892
## 366 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1920
## 367 5.520923e-01 5.520923e-01 Not Significant Desmognathus monticola OTU1925
## 368 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1935
## 369 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1937
## 370 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU1953
## 371 8.744563e-01 8.744563e-01 Not Significant  Desmognathus imitator OTU1956
## 372 7.373227e-01 7.373227e-01 Not Significant  Desmognathus imitator OTU1958
## 373 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1963
## 374 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1964
## 375 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1966
## 376 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1974
## 377 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1978
## 378 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1980
## 379 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1988
## 380 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1991
## 381 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1993
## 382 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1994
## 383 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU1995
## 384 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2002
## 385 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2003
## 386 7.413740e-01 7.413740e-01 Not Significant Desmognathus monticola OTU2008
## 387 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2021
## 388 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2024
## 389 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2025
## 390 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2027
## 391 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2041
## 392 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2045
## 393 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2046
## 394 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2070
## 395 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2071
## 396 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2075
## 397 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2079
## 398 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2093
## 399 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2095
## 400 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2098
## 401 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2102
## 402 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2103
## 403 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2106
## 404 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2107
## 405 6.329740e-02 6.329740e-02 Not Significant  Desmognathus imitator OTU2111
## 406 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2113
## 407 2.943525e-01 2.943525e-01 Not Significant  Desmognathus imitator OTU2131
## 408 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2138
## 409 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2149
## 410 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2152
## 411 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2163
## 412 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2309
## 413 8.944777e-02 8.944777e-02 Not Significant Desmognathus monticola OTU2332
## 414 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2348
## 415 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2352
## 416 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2354
## 417 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2355
## 418 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2357
## 419 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2363
## 420 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2365
## 421 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2366
## 422 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2369
## 423 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2373
## 424 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2376
## 425 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2379
## 426 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2384
## 427 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2386
## 428 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2388
## 429 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2389
## 430 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2391
## 431 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2397
## 432 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2399
## 433 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2400
## 434 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2401
## 435 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2402
## 436 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2409
## 437 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2412
## 438 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2416
## 439 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2426
## 440 8.220190e-02 8.220190e-02 Not Significant Desmognathus monticola OTU2428
## 441 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2431
## 442 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2432
## 443 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2434
## 444 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2435
## 445 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2436
## 446 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2439
## 447 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2444
## 448 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2448
## 449 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2455
## 450 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2462
## 451 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2471
## 452 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2477
## 453 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2478
## 454 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2481
## 455 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2483
## 456 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2485
## 457 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2488
## 458 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2494
## 459 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2497
## 460 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2499
## 461 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2501
## 462 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2502
## 463 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2504
## 464 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2509
## 465 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2511
## 466 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2513
## 467 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2516
## 468 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2527
## 469 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2528
## 470 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2529
## 471 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2530
## 472 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2533
## 473 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2545
## 474 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2547
## 475 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2550
## 476 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2553
## 477 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2555
## 478 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2558
## 479 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2559
## 480 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2560
## 481 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2562
## 482 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2564
## 483 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2569
## 484 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU2577
## 485 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2578
## 486 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2582
## 487 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2584
## 488 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2587
## 489 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2591
## 490 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2593
## 491 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2594
## 492 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2596
## 493 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2602
## 494 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2604
## 495 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2612
## 496 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU2621
## 497 1.424292e-01 1.424292e-01 Not Significant  Desmognathus imitator OTU2672
## 498 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2693
## 499 5.520923e-01 5.520923e-01 Not Significant Desmognathus monticola OTU2702
## 500 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2740
## 501 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2741
## 502 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2745
## 503 2.187349e-01 2.187349e-01 Not Significant  Desmognathus imitator OTU2748
## 504 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2759
## 505 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2762
## 506 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2764
## 507 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2782
## 508 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2784
## 509 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2795
## 510 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2796
## 511 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2818
## 512 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2825
## 513 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2836
## 514 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2840
## 515 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2841
## 516 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2873
## 517 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2876
## 518 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2877
## 519 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2878
## 520 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2884
## 521 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2888
## 522 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2890
## 523 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2891
## 524 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2892
## 525 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2896
## 526 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2899
## 527 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2924
## 528 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU2925
## 529 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU2932
## 530 4.739891e-03 4.739891e-03     Significant  Desmognathus imitator OTU2981
## 531 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3020
## 532 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3033
## 533 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3036
## 534 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3037
## 535 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3038
## 536 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU3039
## 537 3.627526e-02 3.627526e-02 Not Significant  Desmognathus imitator OTU3062
## 538 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3084
## 539 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU3105
## 540 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU3113
## 541 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU3265
## 542 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3312
## 543 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3342
## 544 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3345
## 545 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU3349
## 546 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3372
## 547 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3373
## 548 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3385
## 549 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU3394
## 550 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3396
## 551 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3406
## 552 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3423
## 553 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3426
## 554 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3429
## 555 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3433
## 556 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3442
## 557 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3443
## 558 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3446
## 559 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3449
## 560 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3453
## 561 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3456
## 562 8.643085e-01 8.643085e-01 Not Significant Desmognathus monticola OTU3458
## 563 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3460
## 564 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3461
## 565 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3464
## 566 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3465
## 567 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3467
## 568 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3470
## 569 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3471
## 570 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3472
## 571 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3477
## 572 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3479
## 573 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3480
## 574 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3481
## 575 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3482
## 576 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3484
## 577 5.617051e-12 5.617051e-12     Significant  Desmognathus imitator OTU3485
## 578 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3490
## 579 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3491
## 580 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3493
## 581 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3497
## 582 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3513
## 583 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3516
## 584 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3518
## 585 2.267405e-01 2.267405e-01 Not Significant  Desmognathus imitator OTU3519
## 586 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3520
## 587 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3521
## 588 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3522
## 589 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU3523
## 590 9.419339e-01 9.419339e-01 Not Significant Desmognathus monticola OTU3524
## 591 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3525
## 592 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3526
## 593 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3528
## 594 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3533
## 595 5.465672e-01 5.465672e-01 Not Significant Desmognathus monticola OTU3540
## 596 5.594096e-01 5.594096e-01 Not Significant  Desmognathus imitator OTU3542
## 597 1.592843e-01 1.592843e-01 Not Significant Desmognathus monticola OTU3544
## 598 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3548
## 599 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3572
## 600 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3573
## 601 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3574
## 602 9.395125e-04 9.395125e-04     Significant  Desmognathus imitator OTU3577
## 603 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3578
## 604 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU3603
## 605 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU3605
## 606 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3607
## 607 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU3612
## 608 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3618
## 609 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3640
## 610 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3641
## 611 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3642
## 612 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3643
## 613 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3644
## 614 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3645
## 615 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3646
## 616 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3651
## 617 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3652
## 618 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3654
## 619 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3655
## 620 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3656
## 621 3.556058e-05 3.556058e-05     Significant Desmognathus monticola OTU3658
## 622 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3659
## 623 3.285624e-01 3.285624e-01 Not Significant Desmognathus monticola OTU3662
## 624 8.591374e-01 8.591374e-01 Not Significant  Desmognathus imitator OTU3665
## 625 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3668
## 626 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3673
## 627 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3674
## 628 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3676
## 629 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3684
## 630 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3691
## 631 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU3705
## 632 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU3762
## 633 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3778
## 634 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3837
## 635 5.520923e-01 5.520923e-01 Not Significant Desmognathus monticola OTU3863
## 636 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3866
## 637 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3876
## 638 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU3956
## 639 2.209082e-03 2.209082e-03     Significant Desmognathus monticola OTU3957
## 640 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU3972
## 641 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4033
## 642 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4035
## 643 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4036
## 644 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4037
## 645 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4040
## 646 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU4041
## 647 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4042
## 648 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4043
## 649 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4051
## 650 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4064
## 651 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU4108
## 652 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU4114
## 653 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4121
## 654 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4144
## 655 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4147
## 656 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4149
## 657 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU4173
## 658 7.756834e-03 7.756834e-03     Significant Desmognathus monticola OTU4180
## 659 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4213
## 660 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4219
## 661 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4221
## 662 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4299
## 663 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU4330
## 664 8.926838e-01 8.926838e-01 Not Significant  Desmognathus imitator OTU4336
## 665 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4363
## 666 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4393
## 667 1.614816e-01 1.614816e-01 Not Significant  Desmognathus imitator OTU4426
## 668 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4433
## 669 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU4462
## 670 5.465672e-01 5.465672e-01 Not Significant Desmognathus monticola OTU4508
## 671 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4510
## 672 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4622
## 673 2.943525e-01 2.943525e-01 Not Significant  Desmognathus imitator OTU4630
## 674 7.353263e-01 7.353263e-01 Not Significant Desmognathus monticola OTU4687
## 675 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU4691
## 676 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4711
## 677 4.705693e-02 4.705693e-02 Not Significant  Desmognathus imitator OTU4749
## 678 6.043121e-01 6.043121e-01 Not Significant Desmognathus monticola OTU4770
## 679 1.785236e-02 1.785236e-02 Not Significant Desmognathus monticola OTU4775
## 680 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU4790
## 681 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4945
## 682 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU4954
## 683 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU4984
## 684 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5009
## 685 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5023
## 686 1.842077e-01 1.842077e-01 Not Significant  Desmognathus imitator OTU5030
## 687 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5032
## 688 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU5040
## 689 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU5042
## 690 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5047
## 691 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5061
## 692 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU5063
## 693 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5071
## 694 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5072
## 695 3.183095e-01 3.183095e-01 Not Significant Desmognathus monticola OTU5155
## 696 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5279
## 697 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5293
## 698 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5294
## 699 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5295
## 700 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5296
## 701 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5341
## 702 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5344
## 703 1.944599e-01 1.944599e-01 Not Significant Desmognathus monticola OTU5374
## 704 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5385
## 705 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5388
## 706 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5407
## 707 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5430
## 708 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5431
## 709 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5475
## 710 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5479
## 711 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU5481
## 712 8.220190e-02 8.220190e-02 Not Significant Desmognathus monticola OTU5482
## 713 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5483
## 714 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5484
## 715 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5491
## 716 3.389269e-01 3.389269e-01 Not Significant Desmognathus monticola OTU5500
## 717 1.216979e-06 1.216979e-06     Significant  Desmognathus imitator OTU5505
## 718 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5511
## 719 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5516
## 720 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5524
## 721 7.353263e-01 7.353263e-01 Not Significant Desmognathus monticola OTU5532
## 722 1.074571e-01 1.074571e-01 Not Significant Desmognathus monticola OTU5558
## 723 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5562
## 724 9.520271e-01 9.520271e-01 Not Significant Desmognathus monticola OTU5564
## 725 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5568
## 726 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5569
## 727 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5571
## 728 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5572
## 729 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5574
## 730 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5589
## 731 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5590
## 732 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5642
## 733 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU5646
## 734 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5650
## 735 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU5652
## 736 6.329740e-02 6.329740e-02 Not Significant Desmognathus monticola OTU5664
## 737 5.718205e-01 5.718205e-01 Not Significant Desmognathus monticola OTU5696
## 738 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5701
## 739 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5719
## 740 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5723
## 741 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU5735
## 742 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5743
## 743 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5747
## 744 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5748
## 745 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5753
## 746 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5772
## 747 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5786
## 748 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5787
## 749 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5796
## 750 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5806
## 751 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5817
## 752 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5818
## 753 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5887
## 754 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5958
## 755 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5974
## 756 4.204093e-01 4.204093e-01 Not Significant Desmognathus monticola OTU5976
## 757 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5992
## 758 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU5995
## 759 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6000
## 760 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6009
## 761 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6013
## 762 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU6020
## 763 4.498141e-08 4.498141e-08     Significant Desmognathus monticola OTU6051
## 764 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6081
## 765 3.202247e-01 3.202247e-01 Not Significant Desmognathus monticola OTU6084
## 766 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6095
## 767 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6101
## 768 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6115
## 769 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU6117
## 770 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6120
## 771 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6140
## 772 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6141
## 773 1.388555e-01 1.388555e-01 Not Significant Desmognathus monticola OTU6148
## 774 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU6150
## 775 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6151
## 776 4.176694e-01 4.176694e-01 Not Significant Desmognathus monticola OTU6153
## 777 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6159
## 778 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6162
## 779 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6163
## 780 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6164
## 781 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6165
## 782 1.155414e-01 1.155414e-01 Not Significant Desmognathus monticola OTU6176
## 783 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6188
## 784 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6189
## 785 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6192
## 786 4.817360e-01 4.817360e-01 Not Significant Desmognathus monticola OTU6193
## 787 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6200
## 788 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6232
## 789 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6235
## 790 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6236
## 791 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6238
## 792 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6239
## 793 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6240
## 794 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU6241
## 795 1.592843e-01 1.592843e-01 Not Significant  Desmognathus imitator OTU6274
## 796 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6372
## 797 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6374
## 798 5.718205e-01 5.718205e-01 Not Significant  Desmognathus imitator OTU6383
## 799 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6388
## 800 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6389
## 801 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6390
## 802 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6391
## 803 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6396
## 804 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6401
## 805 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6402
## 806 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6404
## 807 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6405
## 808 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6406
## 809 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6408
## 810 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU6413
## 811 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6414
## 812 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6425
## 813 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6426
## 814 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6429
## 815 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6434
## 816 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6490
## 817 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU6579
## 818 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU6585
## 819 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU6594
## 820 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6600
## 821 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6602
## 822 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6610
## 823 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6617
## 824 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6618
## 825 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6620
## 826 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6624
## 827 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6625
## 828 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6626
## 829 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6629
## 830 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6639
## 831 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6642
## 832 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU6645
## 833 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6647
## 834 2.669231e-01 2.669231e-01 Not Significant  Desmognathus imitator OTU6674
## 835 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6708
## 836 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6709
## 837 3.775877e-03 3.775877e-03     Significant  Desmognathus imitator OTU6713
## 838 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU6750
## 839 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6751
## 840 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6800
## 841 4.142756e-01 4.142756e-01 Not Significant Desmognathus monticola OTU6804
## 842 3.775877e-03 3.775877e-03     Significant  Desmognathus imitator OTU6820
## 843 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6836
## 844 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6840
## 845 8.526314e-01 8.526314e-01 Not Significant  Desmognathus imitator OTU6885
## 846 2.909264e-06 2.909264e-06     Significant  Desmognathus imitator OTU6897
## 847 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6902
## 848 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6903
## 849 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6905
## 850 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6910
## 851 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6913
## 852 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6915
## 853 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU6927
## 854 7.353263e-01 7.353263e-01 Not Significant  Desmognathus imitator OTU6947
## 855 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU6999
## 856 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7000
## 857 8.220190e-02 8.220190e-02 Not Significant Desmognathus monticola OTU7003
## 858 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7004
## 859 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7009
## 860 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7016
## 861 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7019
## 862 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7020
## 863 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7024
## 864 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7026
## 865 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7027
## 866 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7028
## 867 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7029
## 868 1.132275e-01 1.132275e-01 Not Significant Desmognathus monticola OTU7031
## 869 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7032
## 870 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7034
## 871 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7038
## 872 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7040
## 873 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7041
## 874 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7043
## 875 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7044
## 876 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7045
## 877 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7047
## 878 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU7051
## 879 7.987761e-03 7.987761e-03     Significant  Desmognathus imitator OTU7070
## 880 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7078
## 881 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU7091
## 882 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7102
## 883 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7156
## 884 2.997851e-01 2.997851e-01 Not Significant Desmognathus monticola OTU7169
## 885 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7178
## 886 2.930976e-02 2.930976e-02 Not Significant  Desmognathus imitator OTU7189
## 887 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7194
## 888 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7204
## 889 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU7415
## 890 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7436
## 891 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7456
## 892 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7469
## 893 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7470
## 894 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7472
## 895 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7475
## 896 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7484
## 897 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7493
## 898 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7494
## 899 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7496
## 900 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7504
## 901 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7525
## 902 1.155414e-01 1.155414e-01 Not Significant  Desmognathus imitator OTU7527
## 903 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7553
## 904 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7555
## 905 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7558
## 906 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7566
## 907 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7663
## 908 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7665
## 909 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7679
## 910 3.909438e-04 3.909438e-04     Significant Desmognathus monticola OTU7691
## 911 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU7701
## 912 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU7740
## 913 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU7845
## 914 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU7850
## 915 1.241548e-01 1.241548e-01 Not Significant Desmognathus monticola OTU8005
## 916 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU8082
## 917 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU8086
## 918 8.971344e-02 8.971344e-02 Not Significant  Desmognathus imitator OTU8131
## 919 8.724557e-01 8.724557e-01 Not Significant Desmognathus monticola OTU8194
## 920 4.552000e-01 4.552000e-01 Not Significant Desmognathus monticola OTU8256
## 921 7.714710e-02 7.714710e-02 Not Significant  Desmognathus imitator OTU8280
## 922 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU8281
## 923 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU8286
## 924 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU8291
## 925 2.135579e-02 2.135579e-02 Not Significant  Desmognathus imitator OTU8293
## 926 7.353263e-01 7.353263e-01 Not Significant Desmognathus monticola OTU8325
## 927 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU8339
## 928 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU8343
## 929 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU8346
## 930 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU8386
## 931 4.694713e-01 4.694713e-01 Not Significant  Desmognathus imitator OTU8431
## 932 3.202247e-01 3.202247e-01 Not Significant Desmognathus monticola OTU8460
## 933 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU8476
## 934 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU8495
## 935 9.419339e-01 9.419339e-01 Not Significant  Desmognathus imitator OTU8510
## 936 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU8586
## 937 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU8604
## 938 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU8610
## 939 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU8620
## 940 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU8635
## 941 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU8716
## 942 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU8742
## 943 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU8751
## 944 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU8756
## 945 5.681956e-01 5.681956e-01 Not Significant Desmognathus monticola OTU8757
## 946 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU8758
## 947 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU8773
## 948 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU8820
## 949 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU8928
## 950 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU8939
## 951 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU8957
## 952 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU8969
## 953 1.000000e+00 1.000000e+00 Not Significant Desmognathus monticola OTU8977
## 954 1.503214e-01 1.503214e-01 Not Significant Desmognathus monticola OTU8986
## 955 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU9003
## 956 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU9041
## 957 7.353263e-01 7.353263e-01 Not Significant Desmognathus monticola OTU9044
## 958 1.284360e-01 1.284360e-01 Not Significant Desmognathus monticola OTU9075
## 959 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU9184
## 960 1.000000e+00 1.000000e+00 Not Significant  Desmognathus imitator OTU9189
##      Kingdom                       Phylum                        Class
## 1   Bacteria               Armatimonadota                   uncultured
## 2   Bacteria               Proteobacteria          Gammaproteobacteria
## 3   Bacteria                   Firmicutes             Syntrophomonadia
## 4   Bacteria                   Firmicutes              Symbiobacteriia
## 5   Bacteria              Gemmatimonadota             Gemmatimonadetes
## 6   Bacteria              Gemmatimonadota             Gemmatimonadetes
## 7   Bacteria                  Chloroflexi                 Chloroflexia
## 8   Bacteria                  Chloroflexi                 Anaerolineae
## 9   Bacteria                  Chloroflexi                 Chloroflexia
## 10  Bacteria                  Chloroflexi                 Chloroflexia
## 11  Bacteria                  Chloroflexi                 Chloroflexia
## 12  Bacteria                  Chloroflexi                 Anaerolineae
## 13  Bacteria                  Chloroflexi                 Anaerolineae
## 14  Bacteria                  Chloroflexi                 Anaerolineae
## 15  Bacteria                  Chloroflexi                 Anaerolineae
## 16  Bacteria                  Chloroflexi                 Anaerolineae
## 17  Bacteria                  Chloroflexi                 Chloroflexia
## 18  Bacteria                  Chloroflexi              Dehalococcoidia
## 19  Bacteria                  Chloroflexi                 JG30-KF-CM66
## 20  Bacteria                  Chloroflexi                 Anaerolineae
## 21  Bacteria                  Chloroflexi                 Anaerolineae
## 22  Bacteria                  Chloroflexi                 Anaerolineae
## 23  Bacteria                  Chloroflexi              Ktedonobacteria
## 24  Bacteria                  Chloroflexi                 Chloroflexia
## 25  Bacteria                  Chloroflexi                 Chloroflexia
## 26  Bacteria                  Chloroflexi                 Chloroflexia
## 27  Bacteria                  Chloroflexi                 Chloroflexia
## 28  Bacteria                  Chloroflexi                       SHA-26
## 29  Bacteria               Armatimonadota             Chthonomonadetes
## 30  Bacteria                  Chloroflexi                 Chloroflexia
## 31  Bacteria                  Chloroflexi                  Gitt-GS-136
## 32  Bacteria                  Chloroflexi                 Chloroflexia
## 33  Bacteria                  Chloroflexi                        OLB14
## 34  Bacteria                  Chloroflexi              Ktedonobacteria
## 35  Bacteria                  Chloroflexi              Ktedonobacteria
## 36  Bacteria                  Chloroflexi              Ktedonobacteria
## 37  Bacteria                  Chloroflexi              Ktedonobacteria
## 38  Bacteria                  Chloroflexi              Ktedonobacteria
## 39  Bacteria                  Chloroflexi              Ktedonobacteria
## 40  Bacteria                  Chloroflexi              Ktedonobacteria
## 41  Bacteria                  Chloroflexi              Ktedonobacteria
## 42  Bacteria                  Chloroflexi              Ktedonobacteria
## 43  Bacteria            Methylomirabilota             Methylomirabilia
## 44  Bacteria                  Chloroflexi              Ktedonobacteria
## 45  Bacteria                        GAL15                        GAL15
## 46  Bacteria                  Chloroflexi                         TK10
## 47  Bacteria                        WPS-2                        WPS-2
## 48  Bacteria                Spirochaetota                 Brevinematia
## 49  Bacteria              Patescibacteria              Gracilibacteria
## 50  Bacteria                  Chloroflexi                 Anaerolineae
## 51  Bacteria              Acidobacteriota               Blastocatellia
## 52  Bacteria              Acidobacteriota               Blastocatellia
## 53  Bacteria              Acidobacteriota               Blastocatellia
## 54  Bacteria              Acidobacteriota               Blastocatellia
## 55  Bacteria              Elusimicrobiota                  Lineage_IIc
## 56  Bacteria              Elusimicrobiota                  Lineage_IIa
## 57  Bacteria              Elusimicrobiota                  Lineage_IIb
## 58  Bacteria               Proteobacteria          Gammaproteobacteria
## 59  Bacteria              Acidobacteriota               Acidobacteriae
## 60  Bacteria              Acidobacteriota               Acidobacteriae
## 61  Bacteria              Acidobacteriota               Acidobacteriae
## 62  Bacteria                 Nitrospirota                  Nitrospiria
## 63  Bacteria                  Myxococcota                   Myxococcia
## 64  Bacteria              Acidobacteriota               Acidobacteriae
## 65  Bacteria              Acidobacteriota               Acidobacteriae
## 66  Bacteria              Acidobacteriota               Acidobacteriae
## 67  Bacteria              Acidobacteriota               Acidobacteriae
## 68  Bacteria              Acidobacteriota               Acidobacteriae
## 69  Bacteria                  Sumerlaeota                   Sumerlaeia
## 70  Bacteria              Gemmatimonadota                Longimicrobia
## 71  Bacteria              Gemmatimonadota                Longimicrobia
## 72  Bacteria              Acidobacteriota             Vicinamibacteria
## 73  Bacteria                Spirochaetota                       MVP-15
## 74  Bacteria                  Myxococcota                  bacteriap25
## 75  Bacteria SAR324_clade(Marine_group_B) SAR324_clade(Marine_group_B)
## 76  Bacteria                Spirochaetota                 Spirochaetia
## 77  Bacteria                Spirochaetota                 Spirochaetia
## 78  Bacteria                Spirochaetota                 Spirochaetia
## 79  Bacteria             Desulfobacterota                   uncultured
## 80  Bacteria                Spirochaetota                  Leptospirae
## 81  Bacteria                Spirochaetota                  Leptospirae
## 82  Bacteria             Actinobacteriota              Thermoleophilia
## 83  Bacteria             Actinobacteriota              Thermoleophilia
## 84  Bacteria             Actinobacteriota               Acidimicrobiia
## 85  Bacteria             Actinobacteriota               Acidimicrobiia
## 86  Bacteria             Actinobacteriota               Acidimicrobiia
## 87  Bacteria             Actinobacteriota               Acidimicrobiia
## 88  Bacteria             Actinobacteriota               Actinobacteria
## 89  Bacteria             Actinobacteriota               Acidimicrobiia
## 90  Bacteria             Actinobacteriota               Acidimicrobiia
## 91  Bacteria             Actinobacteriota               Acidimicrobiia
## 92  Bacteria             Actinobacteriota               Acidimicrobiia
## 93  Bacteria             Actinobacteriota               Acidimicrobiia
## 94  Bacteria             Actinobacteriota               Acidimicrobiia
## 95  Bacteria             Actinobacteriota               Actinobacteria
## 96  Bacteria             Actinobacteriota               Actinobacteria
## 97  Bacteria             Actinobacteriota               Actinobacteria
## 98  Bacteria             Actinobacteriota               Actinobacteria
## 99  Bacteria             Actinobacteriota               Actinobacteria
## 100 Bacteria                Cyanobacteria               Cyanobacteriia
## 101 Bacteria             Actinobacteriota               Actinobacteria
## 102 Bacteria             Actinobacteriota              Thermoleophilia
## 103 Bacteria             Actinobacteriota              Thermoleophilia
## 104 Bacteria             Actinobacteriota              Thermoleophilia
## 105 Bacteria             Actinobacteriota              Thermoleophilia
## 106 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 107 Bacteria             Actinobacteriota              Thermoleophilia
## 108 Bacteria             Actinobacteriota              Thermoleophilia
## 109 Bacteria             Actinobacteriota              Thermoleophilia
## 110 Bacteria              Acidobacteriota               Acidobacteriae
## 111 Bacteria             Actinobacteriota              Thermoleophilia
## 112 Bacteria              Planctomycetota                    vadinHA49
## 113 Bacteria                 Nitrospirota       Thermodesulfovibrionia
## 114 Bacteria                 Bacteroidota                    Kryptonia
## 115 Bacteria                 Zixibacteria                 Zixibacteria
## 116 Bacteria               Fibrobacterota                Fibrobacteria
## 117 Bacteria               Fibrobacterota                Fibrobacteria
## 118 Bacteria                 Bacteroidota               Ignavibacteria
## 119 Bacteria                 Bacteroidota               Ignavibacteria
## 120 Bacteria                 Bacteroidota                 Kapabacteria
## 121 Bacteria                 Bacteroidota                       SJA-28
## 122 Bacteria                 Bacteroidota                 Rhodothermia
## 123 Bacteria                 Bacteroidota                 Rhodothermia
## 124 Bacteria               Armatimonadota             Chthonomonadetes
## 125 Bacteria               Armatimonadota                Armatimonadia
## 126 Bacteria               Armatimonadota                Armatimonadia
## 127 Bacteria               Proteobacteria          Alphaproteobacteria
## 128 Bacteria               Proteobacteria          Alphaproteobacteria
## 129 Bacteria               Proteobacteria          Alphaproteobacteria
## 130 Bacteria               Proteobacteria          Alphaproteobacteria
## 131 Bacteria               Proteobacteria          Alphaproteobacteria
## 132 Bacteria               Proteobacteria          Alphaproteobacteria
## 133 Bacteria               Proteobacteria          Alphaproteobacteria
## 134 Bacteria               Proteobacteria          Alphaproteobacteria
## 135 Bacteria               Proteobacteria          Alphaproteobacteria
## 136 Bacteria               Proteobacteria          Alphaproteobacteria
## 137 Bacteria               Proteobacteria          Alphaproteobacteria
## 138 Bacteria               Proteobacteria          Alphaproteobacteria
## 139 Bacteria               Proteobacteria          Alphaproteobacteria
## 140 Bacteria               Proteobacteria          Alphaproteobacteria
## 141 Bacteria               Proteobacteria          Alphaproteobacteria
## 142 Bacteria               Proteobacteria          Alphaproteobacteria
## 143 Bacteria               Proteobacteria          Alphaproteobacteria
## 144 Bacteria               Proteobacteria          Gammaproteobacteria
## 145 Bacteria               Proteobacteria          Alphaproteobacteria
## 146 Bacteria               Proteobacteria          Alphaproteobacteria
## 147 Bacteria               Proteobacteria          Alphaproteobacteria
## 148 Bacteria               Proteobacteria          Alphaproteobacteria
## 149 Bacteria               Proteobacteria          Alphaproteobacteria
## 150 Bacteria               Proteobacteria          Alphaproteobacteria
## 151 Bacteria               Proteobacteria          Alphaproteobacteria
## 152 Bacteria               Proteobacteria          Alphaproteobacteria
## 153 Bacteria               Proteobacteria          Alphaproteobacteria
## 154 Bacteria               Proteobacteria          Alphaproteobacteria
## 155 Bacteria               Proteobacteria          Alphaproteobacteria
## 156 Bacteria               Proteobacteria          Alphaproteobacteria
## 157 Bacteria               Proteobacteria          Alphaproteobacteria
## 158 Bacteria               Proteobacteria          Alphaproteobacteria
## 159 Bacteria               Proteobacteria          Alphaproteobacteria
## 160 Bacteria               Proteobacteria          Alphaproteobacteria
## 161 Bacteria               Proteobacteria          Alphaproteobacteria
## 162 Bacteria               Proteobacteria          Alphaproteobacteria
## 163 Bacteria               Proteobacteria          Alphaproteobacteria
## 164 Bacteria               Proteobacteria          Alphaproteobacteria
## 165 Bacteria               Proteobacteria          Alphaproteobacteria
## 166 Bacteria               Proteobacteria          Alphaproteobacteria
## 167 Bacteria               Proteobacteria          Alphaproteobacteria
## 168 Bacteria               Proteobacteria          Alphaproteobacteria
## 169 Bacteria               Proteobacteria          Alphaproteobacteria
## 170 Bacteria               Proteobacteria          Alphaproteobacteria
## 171 Bacteria               Proteobacteria          Alphaproteobacteria
## 172 Bacteria               Proteobacteria          Alphaproteobacteria
## 173 Bacteria               Proteobacteria          Alphaproteobacteria
## 174 Bacteria               Proteobacteria          Alphaproteobacteria
## 175 Bacteria               Proteobacteria          Alphaproteobacteria
## 176 Bacteria               Proteobacteria          Alphaproteobacteria
## 177 Bacteria               Proteobacteria          Alphaproteobacteria
## 178 Bacteria               Proteobacteria          Alphaproteobacteria
## 179 Bacteria               Proteobacteria          Alphaproteobacteria
## 180 Bacteria               Proteobacteria          Alphaproteobacteria
## 181 Bacteria               Proteobacteria          Alphaproteobacteria
## 182 Bacteria               Proteobacteria          Alphaproteobacteria
## 183 Bacteria               Proteobacteria          Alphaproteobacteria
## 184 Bacteria               Proteobacteria          Alphaproteobacteria
## 185 Bacteria               Proteobacteria          Alphaproteobacteria
## 186 Bacteria               Proteobacteria          Alphaproteobacteria
## 187 Bacteria               Proteobacteria          Alphaproteobacteria
## 188 Bacteria               Proteobacteria          Alphaproteobacteria
## 189 Bacteria               Proteobacteria          Alphaproteobacteria
## 190 Bacteria               Proteobacteria          Alphaproteobacteria
## 191 Bacteria               Proteobacteria          Alphaproteobacteria
## 192 Bacteria               Proteobacteria          Alphaproteobacteria
## 193 Bacteria               Proteobacteria          Alphaproteobacteria
## 194 Bacteria               Proteobacteria          Alphaproteobacteria
## 195 Bacteria               Proteobacteria          Alphaproteobacteria
## 196 Bacteria               Proteobacteria          Alphaproteobacteria
## 197 Bacteria               Proteobacteria          Alphaproteobacteria
## 198 Bacteria               Proteobacteria          Alphaproteobacteria
## 199 Bacteria               Proteobacteria          Alphaproteobacteria
## 200 Bacteria               Proteobacteria          Alphaproteobacteria
## 201 Bacteria               Proteobacteria          Alphaproteobacteria
## 202 Bacteria               Proteobacteria          Alphaproteobacteria
## 203 Bacteria               Proteobacteria          Alphaproteobacteria
## 204 Bacteria               Proteobacteria          Alphaproteobacteria
## 205 Bacteria               Proteobacteria          Alphaproteobacteria
## 206 Bacteria               Proteobacteria          Alphaproteobacteria
## 207 Bacteria                Cyanobacteria            Sericytochromatia
## 208 Bacteria               Proteobacteria          Alphaproteobacteria
## 209 Bacteria               Proteobacteria          Gammaproteobacteria
## 210 Bacteria               Proteobacteria          Gammaproteobacteria
## 211 Bacteria               Proteobacteria          Gammaproteobacteria
## 212 Bacteria               Proteobacteria          Gammaproteobacteria
## 213 Bacteria               Fibrobacterota                Fibrobacteria
## 214 Bacteria               Proteobacteria          Gammaproteobacteria
## 215 Bacteria               Proteobacteria          Gammaproteobacteria
## 216 Bacteria               Proteobacteria          Gammaproteobacteria
## 217 Bacteria               Proteobacteria          Gammaproteobacteria
## 218 Bacteria               Proteobacteria          Gammaproteobacteria
## 219 Bacteria               Proteobacteria          Gammaproteobacteria
## 220 Bacteria               Proteobacteria          Gammaproteobacteria
## 221 Bacteria               Proteobacteria          Gammaproteobacteria
## 222 Bacteria               Proteobacteria          Gammaproteobacteria
## 223 Bacteria               Proteobacteria          Gammaproteobacteria
## 224 Bacteria               Proteobacteria          Gammaproteobacteria
## 225 Bacteria               Proteobacteria          Gammaproteobacteria
## 226 Bacteria             Bdellovibrionota              Bdellovibrionia
## 227 Bacteria               Proteobacteria          Gammaproteobacteria
## 228 Bacteria               Proteobacteria          Gammaproteobacteria
## 229 Bacteria               Proteobacteria          Gammaproteobacteria
## 230 Bacteria               Proteobacteria          Gammaproteobacteria
## 231 Bacteria               Proteobacteria          Gammaproteobacteria
## 232 Bacteria               Proteobacteria          Gammaproteobacteria
## 233 Bacteria              Acidobacteriota          Thermoanaerobaculia
## 234 Bacteria               Proteobacteria          Gammaproteobacteria
## 235 Bacteria               Proteobacteria          Gammaproteobacteria
## 236 Bacteria               Proteobacteria          Gammaproteobacteria
## 237 Bacteria               Proteobacteria          Gammaproteobacteria
## 238 Bacteria               Proteobacteria          Gammaproteobacteria
## 239 Bacteria               Proteobacteria          Gammaproteobacteria
## 240 Bacteria               Proteobacteria          Gammaproteobacteria
## 241 Bacteria               Proteobacteria          Gammaproteobacteria
## 242 Bacteria               Proteobacteria          Gammaproteobacteria
## 243 Bacteria                       MBNT15                       MBNT15
## 244 Bacteria                      RCP2-54                      RCP2-54
## 245 Bacteria                 Deinococcota                   Deinococci
## 246 Bacteria                 Deinococcota                   Deinococci
## 247 Bacteria                 Deinococcota                   Deinococci
## 248 Bacteria             Desulfobacterota             Syntrophorhabdia
## 249 Bacteria             Bdellovibrionota              Bdellovibrionia
## 250 Bacteria             Desulfobacterota                   Syntrophia
## 251 Bacteria             Desulfobacterota                   Syntrophia
## 252 Bacteria             Desulfobacterota              Desulfobacteria
## 253 Bacteria             Desulfobacterota             Desulfuromonadia
## 254 Bacteria             Desulfobacterota             Desulfuromonadia
## 255 Bacteria             Desulfobacterota                Desulfobaccia
## 256 Bacteria             Bdellovibrionota                  Oligoflexia
## 257 Bacteria            Methylomirabilota             Methylomirabilia
## 258 Bacteria              Planctomycetota                       BD7-11
## 259 Bacteria                  Myxococcota                    Polyangia
## 260 Bacteria                  Myxococcota                    Polyangia
## 261 Bacteria             Desulfobacterota             Desulfuromonadia
## 262 Bacteria             Bdellovibrionota                  Oligoflexia
## 263 Bacteria             Bdellovibrionota                  Oligoflexia
## 264 Bacteria             Desulfobacterota             Desulfuromonadia
## 265 Bacteria                  Myxococcota                    Polyangia
## 266 Bacteria                        NKB15                        NKB15
## 267 Bacteria               Proteobacteria          Alphaproteobacteria
## 268 Bacteria                  Myxococcota                   Myxococcia
## 269 Bacteria                  Myxococcota                   Myxococcia
## 270 Bacteria                  Myxococcota                   Myxococcia
## 271 Bacteria                  Myxococcota                    Polyangia
## 272 Bacteria                  Myxococcota                    Polyangia
## 273 Bacteria                  Myxococcota                    Polyangia
## 274 Bacteria                  Myxococcota                    Polyangia
## 275 Bacteria                  Myxococcota                    Polyangia
## 276 Bacteria                  Myxococcota                    Polyangia
## 277 Bacteria                  Myxococcota                    Polyangia
## 278 Bacteria                  Myxococcota                    Polyangia
## 279 Bacteria                  Myxococcota                    Polyangia
## 280 Bacteria                  Myxococcota                    Polyangia
## 281 Bacteria                  Myxococcota                    Polyangia
## 282 Bacteria                  Myxococcota                    Polyangia
## 283 Bacteria                        NB1-j                        NB1-j
## 284 Bacteria                  Myxococcota                    Polyangia
## 285 Bacteria                  Myxococcota                    Polyangia
## 286 Bacteria              Acidobacteriota               Acidobacteriae
## 287 Bacteria              Acidobacteriota               Acidobacteriae
## 288 Bacteria              Acidobacteriota                  Subgroup_11
## 289 Bacteria             Latescibacterota             Latescibacterota
## 290 Bacteria              Acidobacteriota               Blastocatellia
## 291 Bacteria              Acidobacteriota               Blastocatellia
## 292 Bacteria              Acidobacteriota               Blastocatellia
## 293 Bacteria              Patescibacteria              Gracilibacteria
## 294 Bacteria              Patescibacteria              Gracilibacteria
## 295  Archaea                Halobacterota              Methanomicrobia
## 296  Archaea                Euryarchaeota              Methanobacteria
## 297  Archaea                Euryarchaeota              Methanobacteria
## 298 Bacteria              Patescibacteria                         ABY1
## 299 Bacteria              Patescibacteria                Parcubacteria
## 300 Bacteria              Patescibacteria                Parcubacteria
## 301 Bacteria              Patescibacteria              Saccharimonadia
## 302 Bacteria               Armatimonadota               Fimbriimonadia
## 303 Bacteria               Armatimonadota               Fimbriimonadia
## 304 Bacteria              Patescibacteria              Saccharimonadia
## 305 Bacteria              Patescibacteria              Saccharimonadia
## 306 Bacteria              Patescibacteria              Saccharimonadia
## 307 Bacteria              Patescibacteria              Saccharimonadia
## 308 Bacteria               Proteobacteria          Gammaproteobacteria
## 309 Bacteria             Campilobacterota              Campylobacteria
## 310 Bacteria            Verrucomicrobiota                  Omnitrophia
## 311 Bacteria              Acidobacteriota             Vicinamibacteria
## 312 Bacteria             Campilobacterota              Campylobacteria
## 313 Bacteria             Campilobacterota              Campylobacteria
## 314 Bacteria             Campilobacterota              Campylobacteria
## 315 Bacteria             Campilobacterota              Campylobacteria
## 316  Archaea                Crenarchaeota              Nitrososphaeria
## 317  Archaea                Crenarchaeota              Nitrososphaeria
## 318  Archaea                Halobacterota              Methanosarcinia
## 319  Archaea                Halobacterota                 Halobacteria
## 320  Archaea                Halobacterota              Methanomicrobia
## 321  Archaea             Thermoplasmatota               Thermoplasmata
## 322  Archaea                Nanoarchaeota                 Nanoarchaeia
## 323 Bacteria                 Dependentiae                     Babeliae
## 324 Bacteria                 Dependentiae                     Babeliae
## 325 Bacteria                 Dependentiae                     Babeliae
## 326 Bacteria                 Dependentiae                     Babeliae
## 327 Bacteria              Acidobacteriota                   Holophagae
## 328 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 329 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 330 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 331 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 332 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 333 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 334 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 335 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 336 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 337 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 338 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 339 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 340 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 341 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 342 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 343 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 344 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 345 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 346 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 347 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 348 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 349 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 350 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 351 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 352 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 353 Bacteria                 Nitrospirota               Leptospirillia
## 354 Bacteria              Acidobacteriota                  Subgroup_22
## 355 Bacteria              Acidobacteriota             Vicinamibacteria
## 356 Bacteria              Acidobacteriota                  Subgroup_18
## 357 Bacteria              Acidobacteriota                     AT-s3-28
## 358 Bacteria              Acidobacteriota                   Holophagae
## 359 Bacteria            Entotheonellaeota               Entotheonellia
## 360 Bacteria                  Myxococcota                   Myxococcia
## 361 Bacteria              Hydrogenedentes              Hydrogenedentia
## 362 Bacteria              Planctomycetota                 Pla4_lineage
## 363 Bacteria              Acidobacteriota          Thermoanaerobaculia
## 364 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 365 Bacteria            Verrucomicrobiota              Kiritimatiellae
## 366 Bacteria              Acidobacteriota               Blastocatellia
## 367 Bacteria                   Firmicutes                      Bacilli
## 368 Bacteria            Verrucomicrobiota                   Chlamydiae
## 369 Bacteria            Verrucomicrobiota                   Chlamydiae
## 370 Bacteria            Verrucomicrobiota                   Chlamydiae
## 371 Bacteria            Verrucomicrobiota                   Chlamydiae
## 372 Bacteria            Verrucomicrobiota                   Chlamydiae
## 373 Bacteria            Verrucomicrobiota                   Chlamydiae
## 374 Bacteria            Verrucomicrobiota                   Chlamydiae
## 375 Bacteria            Verrucomicrobiota                   Chlamydiae
## 376 Bacteria            Verrucomicrobiota                   Chlamydiae
## 377 Bacteria            Verrucomicrobiota                   Chlamydiae
## 378 Bacteria            Verrucomicrobiota                   Chlamydiae
## 379 Bacteria                   Firmicutes                   Clostridia
## 380 Bacteria                   Firmicutes                   Clostridia
## 381 Bacteria                   Firmicutes                   Clostridia
## 382 Bacteria                   Firmicutes                   Clostridia
## 383 Bacteria                   Firmicutes                   Clostridia
## 384 Bacteria                 Synergistota                  Synergistia
## 385 Bacteria                 Synergistota                  Synergistia
## 386 Bacteria                   Firmicutes                   Clostridia
## 387 Bacteria             Actinobacteriota               Coriobacteriia
## 388 Bacteria             Actinobacteriota               Coriobacteriia
## 389 Bacteria             Actinobacteriota               Coriobacteriia
## 390 Bacteria             Actinobacteriota               Coriobacteriia
## 391 Bacteria               Proteobacteria          Gammaproteobacteria
## 392 Bacteria               Proteobacteria          Gammaproteobacteria
## 393 Bacteria               Proteobacteria          Gammaproteobacteria
## 394 Bacteria               Proteobacteria          Gammaproteobacteria
## 395 Bacteria               Proteobacteria          Gammaproteobacteria
## 396 Bacteria               Proteobacteria          Gammaproteobacteria
## 397 Bacteria               Proteobacteria          Gammaproteobacteria
## 398 Bacteria                      FCPU426                      FCPU426
## 399 Bacteria               Proteobacteria          Gammaproteobacteria
## 400 Bacteria                   Firmicutes                      Bacilli
## 401 Bacteria                   Firmicutes                      Bacilli
## 402 Bacteria                   Firmicutes                      Bacilli
## 403 Bacteria            Methylomirabilota             Methylomirabilia
## 404 Bacteria            Methylomirabilota             Methylomirabilia
## 405 Bacteria                  Chloroflexi                          AD3
## 406 Bacteria                 Dadabacteria                Dadabacteriia
## 407 Bacteria                 Bacteroidota                  Bacteroidia
## 408 Bacteria                 Bacteroidota                  Bacteroidia
## 409 Bacteria                 Bacteroidota                  Bacteroidia
## 410 Bacteria                 Bacteroidota                  Bacteroidia
## 411 Bacteria                 Bacteroidota                  Bacteroidia
## 412 Bacteria                   Firmicutes                   Clostridia
## 413 Bacteria                 Bacteroidota                  Bacteroidia
## 414 Bacteria                 Bacteroidota                  Bacteroidia
## 415 Bacteria                 Bacteroidota                  Bacteroidia
## 416 Bacteria                 Bacteroidota                  Bacteroidia
## 417 Bacteria                 Bacteroidota                  Bacteroidia
## 418 Bacteria                 Bacteroidota                  Bacteroidia
## 419 Bacteria                 Bacteroidota                  Bacteroidia
## 420 Bacteria                 Bacteroidota                  Bacteroidia
## 421 Bacteria                 Bacteroidota                  Bacteroidia
## 422 Bacteria                 Bacteroidota                  Bacteroidia
## 423 Bacteria                 Bacteroidota                  Bacteroidia
## 424 Bacteria                 Bacteroidota                  Bacteroidia
## 425 Bacteria                 Bacteroidota                  Bacteroidia
## 426 Bacteria                 Bacteroidota                  Bacteroidia
## 427 Bacteria                 Bacteroidota                  Bacteroidia
## 428 Bacteria                 Bacteroidota                  Bacteroidia
## 429 Bacteria                 Bacteroidota                  Bacteroidia
## 430 Bacteria                 Bacteroidota                  Bacteroidia
## 431 Bacteria                 Bacteroidota                  Bacteroidia
## 432 Bacteria                 Bacteroidota                  Bacteroidia
## 433 Bacteria                 Bacteroidota                  Bacteroidia
## 434 Bacteria                 Bacteroidota                  Bacteroidia
## 435 Bacteria                 Bacteroidota                  Bacteroidia
## 436 Bacteria                 Bacteroidota                  Bacteroidia
## 437 Bacteria                 Bacteroidota                  Bacteroidia
## 438 Bacteria                 Bacteroidota                  Bacteroidia
## 439 Bacteria                 Bacteroidota                  Bacteroidia
## 440 Bacteria                 Bacteroidota                  Bacteroidia
## 441 Bacteria                 Bacteroidota                  Bacteroidia
## 442 Bacteria                 Bacteroidota                  Bacteroidia
## 443 Bacteria                 Bacteroidota                  Bacteroidia
## 444 Bacteria                 Bacteroidota                  Bacteroidia
## 445 Bacteria                 Bacteroidota                  Bacteroidia
## 446 Bacteria                 Bacteroidota                  Bacteroidia
## 447 Bacteria                 Bacteroidota                  Bacteroidia
## 448 Bacteria                 Bacteroidota                  Bacteroidia
## 449 Bacteria                 Bacteroidota                  Bacteroidia
## 450 Bacteria                 Bacteroidota                  Bacteroidia
## 451 Bacteria                 Bacteroidota                  Bacteroidia
## 452 Bacteria                 Bacteroidota                  Bacteroidia
## 453 Bacteria                 Bacteroidota                  Bacteroidia
## 454 Bacteria                 Bacteroidota                  Bacteroidia
## 455 Bacteria                 Bacteroidota                  Bacteroidia
## 456 Bacteria                 Bacteroidota                  Bacteroidia
## 457 Bacteria                 Bacteroidota                  Bacteroidia
## 458 Bacteria                 Bacteroidota                  Bacteroidia
## 459 Bacteria                 Bacteroidota                  Bacteroidia
## 460 Bacteria                 Bacteroidota                  Bacteroidia
## 461 Bacteria                 Bacteroidota                  Bacteroidia
## 462 Bacteria                 Bacteroidota                  Bacteroidia
## 463 Bacteria                 Bacteroidota                  Bacteroidia
## 464 Bacteria                 Bacteroidota                  Bacteroidia
## 465 Bacteria                 Bacteroidota                  Bacteroidia
## 466 Bacteria                 Bacteroidota                  Bacteroidia
## 467 Bacteria                 Bacteroidota                  Bacteroidia
## 468 Bacteria                 Bacteroidota                  Bacteroidia
## 469 Bacteria                 Bacteroidota                  Bacteroidia
## 470 Bacteria                 Bacteroidota                  Bacteroidia
## 471 Bacteria                 Bacteroidota                  Bacteroidia
## 472 Bacteria                 Bacteroidota                  Bacteroidia
## 473 Bacteria                 Bacteroidota                  Bacteroidia
## 474 Bacteria                 Bacteroidota                  Bacteroidia
## 475 Bacteria                 Bacteroidota                  Bacteroidia
## 476 Bacteria                 Bacteroidota                  Bacteroidia
## 477 Bacteria                 Bacteroidota                  Bacteroidia
## 478 Bacteria                 Bacteroidota                  Bacteroidia
## 479 Bacteria                 Bacteroidota                  Bacteroidia
## 480 Bacteria                 Bacteroidota                  Bacteroidia
## 481 Bacteria                 Bacteroidota                  Bacteroidia
## 482 Bacteria                 Bacteroidota                  Bacteroidia
## 483 Bacteria                 Bacteroidota                  Bacteroidia
## 484 Bacteria                 Bacteroidota                  Bacteroidia
## 485 Bacteria                 Bacteroidota                  Bacteroidia
## 486 Bacteria                 Bacteroidota                  Bacteroidia
## 487 Bacteria                 Bacteroidota                  Bacteroidia
## 488 Bacteria                 Bacteroidota                  Bacteroidia
## 489 Bacteria                 Bacteroidota                  Bacteroidia
## 490 Bacteria                 Bacteroidota                  Bacteroidia
## 491 Bacteria                 Bacteroidota                  Bacteroidia
## 492 Bacteria                 Bacteroidota                  Bacteroidia
## 493 Bacteria                 Bacteroidota                  Bacteroidia
## 494 Bacteria                 Bacteroidota                  Bacteroidia
## 495 Bacteria                 Bacteroidota                  Bacteroidia
## 496 Bacteria                 Bacteroidota                  Bacteroidia
## 497 Bacteria              Planctomycetota               Planctomycetes
## 498 Bacteria              Planctomycetota               Planctomycetes
## 499 Bacteria              Planctomycetota               Planctomycetes
## 500 Bacteria              Planctomycetota               Planctomycetes
## 501 Bacteria              Planctomycetota               Planctomycetes
## 502 Bacteria              Planctomycetota               Planctomycetes
## 503 Bacteria              Planctomycetota               Planctomycetes
## 504 Bacteria              Planctomycetota               Planctomycetes
## 505 Bacteria              Planctomycetota               Planctomycetes
## 506 Bacteria              Planctomycetota               Planctomycetes
## 507 Bacteria              Planctomycetota               Planctomycetes
## 508 Bacteria              Planctomycetota               Planctomycetes
## 509 Bacteria              Planctomycetota               Planctomycetes
## 510 Bacteria              Planctomycetota               Planctomycetes
## 511 Bacteria              Planctomycetota               Planctomycetes
## 512 Bacteria              Planctomycetota               Planctomycetes
## 513 Bacteria              Planctomycetota               Planctomycetes
## 514 Bacteria              Planctomycetota               Planctomycetes
## 515 Bacteria              Planctomycetota               Planctomycetes
## 516 Bacteria              Planctomycetota                        OM190
## 517 Bacteria              Planctomycetota                Phycisphaerae
## 518 Bacteria              Planctomycetota                Phycisphaerae
## 519 Bacteria              Planctomycetota                Phycisphaerae
## 520 Bacteria              Planctomycetota                Phycisphaerae
## 521 Bacteria              Planctomycetota                Phycisphaerae
## 522 Bacteria              Planctomycetota                Phycisphaerae
## 523 Bacteria              Planctomycetota                Phycisphaerae
## 524 Bacteria              Planctomycetota                Phycisphaerae
## 525 Bacteria              Planctomycetota                Phycisphaerae
## 526 Bacteria              Planctomycetota                Phycisphaerae
## 527 Bacteria              Planctomycetota               Planctomycetes
## 528 Bacteria              Planctomycetota               Planctomycetes
## 529 Bacteria              Planctomycetota               Planctomycetes
## 530 Bacteria              Planctomycetota               Planctomycetes
## 531 Bacteria              Planctomycetota               Planctomycetes
## 532 Bacteria              Planctomycetota               Planctomycetes
## 533 Bacteria              Planctomycetota               Planctomycetes
## 534 Bacteria              Planctomycetota               Planctomycetes
## 535 Bacteria              Planctomycetota               Planctomycetes
## 536 Bacteria              Planctomycetota               Planctomycetes
## 537 Bacteria              Planctomycetota               Planctomycetes
## 538 Bacteria              Patescibacteria                Parcubacteria
## 539 Bacteria             Desulfobacterota             Desulfovibrionia
## 540 Bacteria             Desulfobacterota             Desulfovibrionia
## 541 Bacteria              Acidobacteriota                   Subgroup_5
## 542 Bacteria                   Firmicutes                   Clostridia
## 543 Bacteria                   Firmicutes                   Clostridia
## 544 Bacteria                   Firmicutes                   Clostridia
## 545 Bacteria                   Firmicutes                   Clostridia
## 546 Bacteria                   Firmicutes                      Bacilli
## 547 Bacteria                   Firmicutes                      Bacilli
## 548 Bacteria             Actinobacteriota                Rubrobacteria
## 549 Bacteria             Actinobacteriota                    MB-A2-108
## 550 Bacteria             Actinobacteriota               Actinobacteria
## 551 Bacteria             Actinobacteriota               Actinobacteria
## 552 Bacteria             Actinobacteriota               Actinobacteria
## 553 Bacteria             Actinobacteriota               Actinobacteria
## 554 Bacteria             Actinobacteriota               Actinobacteria
## 555 Bacteria             Actinobacteriota               Actinobacteria
## 556 Bacteria             Actinobacteriota               Actinobacteria
## 557 Bacteria             Actinobacteriota               Actinobacteria
## 558 Bacteria             Actinobacteriota               Actinobacteria
## 559 Bacteria             Actinobacteriota               Actinobacteria
## 560 Bacteria             Actinobacteriota               Actinobacteria
## 561 Bacteria             Actinobacteriota               Actinobacteria
## 562 Bacteria             Actinobacteriota               Actinobacteria
## 563 Bacteria             Actinobacteriota               Actinobacteria
## 564 Bacteria             Actinobacteriota               Actinobacteria
## 565 Bacteria             Actinobacteriota               Actinobacteria
## 566 Bacteria             Actinobacteriota               Actinobacteria
## 567 Bacteria             Actinobacteriota               Actinobacteria
## 568 Bacteria             Actinobacteriota               Actinobacteria
## 569 Bacteria             Actinobacteriota               Actinobacteria
## 570 Bacteria             Actinobacteriota               Actinobacteria
## 571 Bacteria             Actinobacteriota               Actinobacteria
## 572 Bacteria             Actinobacteriota               Actinobacteria
## 573 Bacteria             Actinobacteriota               Actinobacteria
## 574 Bacteria             Actinobacteriota               Actinobacteria
## 575 Bacteria             Actinobacteriota               Actinobacteria
## 576 Bacteria             Actinobacteriota               Actinobacteria
## 577 Bacteria             Actinobacteriota               Actinobacteria
## 578 Bacteria             Actinobacteriota               Actinobacteria
## 579 Bacteria             Actinobacteriota               Actinobacteria
## 580 Bacteria             Actinobacteriota               Actinobacteria
## 581 Bacteria             Actinobacteriota               Actinobacteria
## 582 Bacteria             Actinobacteriota               Actinobacteria
## 583 Bacteria             Actinobacteriota               Actinobacteria
## 584 Bacteria             Actinobacteriota               Actinobacteria
## 585 Bacteria             Actinobacteriota               Actinobacteria
## 586 Bacteria             Actinobacteriota               Actinobacteria
## 587 Bacteria             Actinobacteriota               Actinobacteria
## 588 Bacteria             Actinobacteriota               Actinobacteria
## 589 Bacteria             Actinobacteriota               Actinobacteria
## 590 Bacteria             Actinobacteriota               Actinobacteria
## 591 Bacteria             Actinobacteriota               Actinobacteria
## 592 Bacteria             Actinobacteriota               Actinobacteria
## 593 Bacteria             Actinobacteriota               Actinobacteria
## 594 Bacteria             Actinobacteriota               Actinobacteria
## 595 Bacteria             Actinobacteriota               Actinobacteria
## 596 Bacteria             Actinobacteriota               Actinobacteria
## 597 Bacteria             Actinobacteriota               Actinobacteria
## 598 Bacteria             Actinobacteriota               Actinobacteria
## 599 Bacteria             Actinobacteriota               Actinobacteria
## 600 Bacteria             Actinobacteriota               Actinobacteria
## 601 Bacteria             Actinobacteriota               Actinobacteria
## 602 Bacteria             Actinobacteriota               Actinobacteria
## 603 Bacteria             Actinobacteriota               Actinobacteria
## 604 Bacteria             Actinobacteriota               Actinobacteria
## 605 Bacteria             Actinobacteriota               Actinobacteria
## 606 Bacteria             Actinobacteriota               Actinobacteria
## 607 Bacteria             Actinobacteriota               Actinobacteria
## 608 Bacteria             Actinobacteriota               Actinobacteria
## 609 Bacteria               Proteobacteria          Gammaproteobacteria
## 610 Bacteria               Proteobacteria          Gammaproteobacteria
## 611 Bacteria               Proteobacteria          Gammaproteobacteria
## 612 Bacteria               Proteobacteria          Gammaproteobacteria
## 613 Bacteria              Acidobacteriota                   Holophagae
## 614 Bacteria               Proteobacteria          Gammaproteobacteria
## 615 Bacteria               Proteobacteria          Gammaproteobacteria
## 616 Bacteria               Proteobacteria          Gammaproteobacteria
## 617 Bacteria               Proteobacteria          Gammaproteobacteria
## 618 Bacteria               Proteobacteria          Gammaproteobacteria
## 619 Bacteria               Proteobacteria          Gammaproteobacteria
## 620 Bacteria               Proteobacteria          Gammaproteobacteria
## 621 Bacteria               Proteobacteria          Gammaproteobacteria
## 622 Bacteria               Proteobacteria          Gammaproteobacteria
## 623 Bacteria               Proteobacteria          Gammaproteobacteria
## 624 Bacteria               Proteobacteria          Gammaproteobacteria
## 625 Bacteria               Proteobacteria          Gammaproteobacteria
## 626 Bacteria               Proteobacteria          Gammaproteobacteria
## 627 Bacteria               Proteobacteria          Gammaproteobacteria
## 628 Bacteria               Proteobacteria          Gammaproteobacteria
## 629 Bacteria                   Firmicutes                      Bacilli
## 630 Bacteria               Proteobacteria          Gammaproteobacteria
## 631 Bacteria               Proteobacteria          Alphaproteobacteria
## 632 Bacteria                   Firmicutes                   Clostridia
## 633 Bacteria                   Firmicutes                   Clostridia
## 634 Bacteria                   Firmicutes                   Clostridia
## 635 Bacteria                   Firmicutes                   Clostridia
## 636 Bacteria                   Firmicutes                   Clostridia
## 637 Bacteria                   Firmicutes                   Clostridia
## 638 Bacteria                   Firmicutes                   Clostridia
## 639 Bacteria                   Firmicutes                   Clostridia
## 640 Bacteria                   Firmicutes                   Clostridia
## 641 Bacteria                   Firmicutes                   Clostridia
## 642 Bacteria                   Firmicutes                   Clostridia
## 643 Bacteria                   Firmicutes                   Clostridia
## 644 Bacteria                   Firmicutes                   Clostridia
## 645 Bacteria                   Firmicutes                   Clostridia
## 646 Bacteria                   Firmicutes                   Clostridia
## 647 Bacteria                   Firmicutes                   Clostridia
## 648 Bacteria                   Firmicutes                   Clostridia
## 649 Bacteria                   Firmicutes                   Clostridia
## 650 Bacteria                   Firmicutes                   Clostridia
## 651 Bacteria                   Firmicutes                   Clostridia
## 652 Bacteria                   Firmicutes                   Clostridia
## 653 Bacteria                   Firmicutes                   Clostridia
## 654 Bacteria                   Firmicutes                   Clostridia
## 655 Bacteria                   Firmicutes                   Clostridia
## 656 Bacteria                   Firmicutes                   Clostridia
## 657 Bacteria                   Firmicutes                   Clostridia
## 658 Bacteria                   Firmicutes                   Clostridia
## 659 Bacteria             Bdellovibrionota              Bdellovibrionia
## 660 Bacteria             Bdellovibrionota              Bdellovibrionia
## 661 Bacteria             Abditibacteriota               Abditibacteria
## 662 Bacteria                   Firmicutes                   Clostridia
## 663 Bacteria                   Firmicutes                   Clostridia
## 664 Bacteria                   Firmicutes                   Clostridia
## 665 Bacteria                   Firmicutes                   Clostridia
## 666 Bacteria                   Firmicutes                   Clostridia
## 667 Bacteria                   Firmicutes                   Clostridia
## 668 Bacteria                   Firmicutes                   Clostridia
## 669 Bacteria                   Firmicutes                   Clostridia
## 670 Bacteria                   Firmicutes                   Clostridia
## 671 Bacteria                   Firmicutes                   Clostridia
## 672 Bacteria                   Firmicutes                   Clostridia
## 673 Bacteria                   Firmicutes                   Clostridia
## 674 Bacteria                   Firmicutes                   Clostridia
## 675 Bacteria                   Firmicutes                   Clostridia
## 676 Bacteria                   Firmicutes                   Clostridia
## 677 Bacteria                   Firmicutes                   Clostridia
## 678 Bacteria                   Firmicutes                   Clostridia
## 679 Bacteria                   Firmicutes                   Clostridia
## 680 Bacteria                   Firmicutes                   Clostridia
## 681 Bacteria                   Firmicutes                   Clostridia
## 682 Bacteria                   Firmicutes                   Clostridia
## 683 Bacteria                   Firmicutes                   Clostridia
## 684 Bacteria                   Firmicutes           Desulfitobacteriia
## 685 Bacteria                   Firmicutes             Desulfotomaculia
## 686 Bacteria                   Firmicutes             Desulfotomaculia
## 687 Bacteria                   Firmicutes                   Clostridia
## 688 Bacteria                   Firmicutes           Desulfitobacteriia
## 689 Bacteria                   Firmicutes           Desulfitobacteriia
## 690 Bacteria                   Firmicutes                   Clostridia
## 691 Bacteria                   Firmicutes                   Clostridia
## 692 Bacteria                   Firmicutes                   Clostridia
## 693 Bacteria                   Firmicutes                Negativicutes
## 694 Bacteria                   Firmicutes                     BRH-c20a
## 695 Bacteria                   Firmicutes                   Clostridia
## 696 Bacteria                 Bacteroidota                  Bacteroidia
## 697 Bacteria              Planctomycetota                Phycisphaerae
## 698 Bacteria                 Bacteroidota                  Bacteroidia
## 699 Bacteria                 Bacteroidota                  Bacteroidia
## 700 Bacteria                 Bacteroidota                  Bacteroidia
## 701 Bacteria                 Bacteroidota                  Bacteroidia
## 702 Bacteria               Proteobacteria          Alphaproteobacteria
## 703 Bacteria                   Firmicutes                   Clostridia
## 704 Bacteria                   Firmicutes                   Clostridia
## 705 Bacteria                   Firmicutes                   Clostridia
## 706 Bacteria                   Firmicutes                   Clostridia
## 707 Bacteria                   Firmicutes                   Clostridia
## 708 Bacteria                   Firmicutes                   Clostridia
## 709 Bacteria                   Firmicutes                   Clostridia
## 710 Bacteria                   Firmicutes                   Clostridia
## 711 Bacteria                   Firmicutes                   Clostridia
## 712 Bacteria                   Firmicutes                   Clostridia
## 713 Bacteria                   Firmicutes                   Clostridia
## 714 Bacteria                   Firmicutes                   Clostridia
## 715 Bacteria                   Firmicutes                   Clostridia
## 716 Bacteria                   Firmicutes                   Clostridia
## 717 Bacteria                   Firmicutes                   Clostridia
## 718 Bacteria                   Firmicutes                   Clostridia
## 719 Bacteria             Margulisbacteria             Margulisbacteria
## 720 Bacteria                   Firmicutes               Incertae_Sedis
## 721 Bacteria                   Firmicutes                   Clostridia
## 722 Bacteria                   Firmicutes                   Clostridia
## 723 Bacteria                   Firmicutes                   Clostridia
## 724 Bacteria                   Firmicutes                   Clostridia
## 725 Bacteria                   Firmicutes                      Bacilli
## 726 Bacteria                   Firmicutes                      Bacilli
## 727 Bacteria                   Firmicutes               Natranaerobiia
## 728 Bacteria                   Firmicutes                      Bacilli
## 729 Bacteria                   Firmicutes                   Clostridia
## 730 Bacteria                   Firmicutes                      Bacilli
## 731 Bacteria                   Firmicutes                      Bacilli
## 732 Bacteria                   Firmicutes                   Clostridia
## 733 Bacteria                   Firmicutes                   Clostridia
## 734 Bacteria                   Firmicutes                   Clostridia
## 735 Bacteria                   Firmicutes                   Clostridia
## 736 Bacteria                   Firmicutes                   Clostridia
## 737 Bacteria                   Firmicutes                   Clostridia
## 738 Bacteria                   Firmicutes                   Clostridia
## 739 Bacteria                   Firmicutes                   Clostridia
## 740 Bacteria                   Firmicutes                   Clostridia
## 741 Bacteria                   Firmicutes                 Limnochordia
## 742 Bacteria                   Firmicutes                   Clostridia
## 743 Bacteria                   Firmicutes                   Clostridia
## 744 Bacteria                   Firmicutes             Desulfotomaculia
## 745 Bacteria                   Firmicutes                   Clostridia
## 746 Bacteria                  Myxococcota                    Polyangia
## 747 Bacteria                 Nitrospinota                    P9X2b3D02
## 748 Bacteria             Campilobacterota              Campylobacteria
## 749 Bacteria               Proteobacteria          Alphaproteobacteria
## 750 Bacteria               Proteobacteria          Alphaproteobacteria
## 751 Bacteria               Proteobacteria          Gammaproteobacteria
## 752 Bacteria               Proteobacteria          Gammaproteobacteria
## 753 Bacteria                   Firmicutes                   Clostridia
## 754 Bacteria                   Firmicutes           Desulfitobacteriia
## 755 Bacteria                   Firmicutes                   Clostridia
## 756 Bacteria                   Firmicutes                   Clostridia
## 757 Bacteria                   Firmicutes                   Clostridia
## 758 Bacteria                   Firmicutes                      Bacilli
## 759 Bacteria                   Firmicutes                      Bacilli
## 760 Bacteria                   Firmicutes                   Clostridia
## 761 Bacteria                   Firmicutes                   Clostridia
## 762 Bacteria                   Firmicutes                   Clostridia
## 763 Bacteria                   Firmicutes                   Clostridia
## 764 Bacteria                   Firmicutes                      Bacilli
## 765 Bacteria                   Firmicutes                      Bacilli
## 766 Bacteria             Actinobacteriota               Coriobacteriia
## 767 Bacteria             Actinobacteriota               Coriobacteriia
## 768 Bacteria             Actinobacteriota               Coriobacteriia
## 769 Bacteria             Actinobacteriota               Coriobacteriia
## 770 Bacteria             Actinobacteriota               Coriobacteriia
## 771 Bacteria                   Firmicutes                   Clostridia
## 772 Bacteria                   Firmicutes                Negativicutes
## 773 Bacteria                   Firmicutes                Negativicutes
## 774 Bacteria                   Firmicutes                Negativicutes
## 775 Bacteria                   Firmicutes                Negativicutes
## 776 Bacteria                   Firmicutes                Negativicutes
## 777 Bacteria                   Firmicutes                Negativicutes
## 778 Bacteria                   Firmicutes                Negativicutes
## 779 Bacteria                   Firmicutes           Desulfitobacteriia
## 780 Bacteria                   Firmicutes                      Bacilli
## 781 Bacteria                   Firmicutes                      Bacilli
## 782 Bacteria            Verrucomicrobiota                Lentisphaeria
## 783 Bacteria                  Chloroflexi                 Anaerolineae
## 784 Bacteria                   Firmicutes                      Bacilli
## 785 Bacteria                   Firmicutes                      Bacilli
## 786 Bacteria                   Firmicutes                      Bacilli
## 787 Bacteria                   Firmicutes                      Bacilli
## 788 Bacteria                   Firmicutes                      Bacilli
## 789 Bacteria                   Firmicutes                      Bacilli
## 790 Bacteria                   Firmicutes                      Bacilli
## 791 Bacteria                   Firmicutes                      Bacilli
## 792 Bacteria                   Firmicutes                      Bacilli
## 793 Bacteria                   Firmicutes                      Bacilli
## 794 Bacteria               Proteobacteria          Gammaproteobacteria
## 795 Bacteria                   Firmicutes                      Bacilli
## 796 Bacteria                Cyanobacteria             Vampirivibrionia
## 797 Bacteria                Cyanobacteria             Vampirivibrionia
## 798 Bacteria                Cyanobacteria             Vampirivibrionia
## 799 Bacteria                Cyanobacteria               Cyanobacteriia
## 800 Bacteria                Cyanobacteria               Cyanobacteriia
## 801 Bacteria                Cyanobacteria               Cyanobacteriia
## 802 Bacteria                Cyanobacteria               Cyanobacteriia
## 803 Bacteria                Cyanobacteria               Cyanobacteriia
## 804 Bacteria                Cyanobacteria               Cyanobacteriia
## 805 Bacteria                Cyanobacteria               Cyanobacteriia
## 806 Bacteria                Cyanobacteria               Cyanobacteriia
## 807 Bacteria                Cyanobacteria               Cyanobacteriia
## 808 Bacteria                Cyanobacteria               Cyanobacteriia
## 809 Bacteria                Cyanobacteria               Cyanobacteriia
## 810 Bacteria                Cyanobacteria               Cyanobacteriia
## 811 Bacteria                Cyanobacteria               Cyanobacteriia
## 812 Bacteria                Cyanobacteria               Cyanobacteriia
## 813 Bacteria                Cyanobacteria               Cyanobacteriia
## 814 Bacteria                Cyanobacteria               Cyanobacteriia
## 815 Bacteria                Cyanobacteria               Cyanobacteriia
## 816 Bacteria                   Firmicutes                      Bacilli
## 817 Bacteria                   Firmicutes                      Bacilli
## 818 Bacteria                   Firmicutes                      Bacilli
## 819 Bacteria                   Firmicutes                      Bacilli
## 820 Bacteria                   Firmicutes                      Bacilli
## 821 Bacteria                   Firmicutes                      Bacilli
## 822 Bacteria                   Firmicutes                      Bacilli
## 823 Bacteria                   Firmicutes                      Bacilli
## 824 Bacteria                   Firmicutes                      Bacilli
## 825 Bacteria                   Firmicutes                      Bacilli
## 826 Bacteria                   Firmicutes                      Bacilli
## 827 Bacteria                   Firmicutes                      Bacilli
## 828 Bacteria                   Firmicutes                      Bacilli
## 829 Bacteria                   Firmicutes                      Bacilli
## 830 Bacteria                   Firmicutes                      Bacilli
## 831 Bacteria                   Firmicutes                      Bacilli
## 832 Bacteria                   Firmicutes                      Bacilli
## 833 Bacteria                   Firmicutes                      Bacilli
## 834 Bacteria                   Firmicutes                      Bacilli
## 835 Bacteria                   Firmicutes                      Bacilli
## 836 Bacteria                   Firmicutes                      Bacilli
## 837 Bacteria                   Firmicutes                      Bacilli
## 838 Bacteria                   Firmicutes                      Bacilli
## 839 Bacteria                   Firmicutes                      Bacilli
## 840 Bacteria                   Firmicutes                   Clostridia
## 841 Bacteria                   Firmicutes                   Clostridia
## 842 Bacteria             Deferribacterota              Deferribacteres
## 843 Bacteria                 Synergistota                  Synergistia
## 844 Bacteria                 Synergistota                  Synergistia
## 845 Bacteria                  Myxococcota                   Myxococcia
## 846 Bacteria                   Firmicutes                      Bacilli
## 847 Bacteria                   Firmicutes                      Bacilli
## 848 Bacteria               Proteobacteria          Gammaproteobacteria
## 849 Bacteria               Proteobacteria          Gammaproteobacteria
## 850 Bacteria               Proteobacteria          Gammaproteobacteria
## 851 Bacteria               Proteobacteria          Gammaproteobacteria
## 852 Bacteria               Proteobacteria          Gammaproteobacteria
## 853 Bacteria             Desulfobacterota             Desulfovibrionia
## 854 Bacteria             Actinobacteriota               Acidimicrobiia
## 855 Bacteria               Proteobacteria          Gammaproteobacteria
## 856 Bacteria               Proteobacteria          Gammaproteobacteria
## 857 Bacteria               Proteobacteria          Gammaproteobacteria
## 858 Bacteria               Proteobacteria          Gammaproteobacteria
## 859 Bacteria               Proteobacteria          Gammaproteobacteria
## 860 Bacteria               Proteobacteria          Gammaproteobacteria
## 861 Bacteria               Proteobacteria          Gammaproteobacteria
## 862 Bacteria               Proteobacteria          Gammaproteobacteria
## 863 Bacteria               Proteobacteria          Gammaproteobacteria
## 864 Bacteria               Proteobacteria          Gammaproteobacteria
## 865 Bacteria               Proteobacteria          Gammaproteobacteria
## 866 Bacteria               Proteobacteria          Gammaproteobacteria
## 867 Bacteria               Proteobacteria          Gammaproteobacteria
## 868 Bacteria               Proteobacteria          Gammaproteobacteria
## 869 Bacteria               Proteobacteria          Gammaproteobacteria
## 870 Bacteria               Proteobacteria          Gammaproteobacteria
## 871 Bacteria               Proteobacteria          Gammaproteobacteria
## 872 Bacteria               Proteobacteria          Gammaproteobacteria
## 873 Bacteria               Proteobacteria          Gammaproteobacteria
## 874 Bacteria               Proteobacteria          Gammaproteobacteria
## 875 Bacteria               Proteobacteria          Gammaproteobacteria
## 876 Bacteria               Proteobacteria          Gammaproteobacteria
## 877 Bacteria               Proteobacteria          Gammaproteobacteria
## 878 Bacteria               Proteobacteria          Gammaproteobacteria
## 879 Bacteria             Actinobacteriota               Actinobacteria
## 880 Bacteria             Actinobacteriota               Actinobacteria
## 881 Bacteria                   Firmicutes                   Clostridia
## 882 Bacteria                   Firmicutes                   Clostridia
## 883 Bacteria            Verrucomicrobiota                   Chlamydiae
## 884 Bacteria              Acidobacteriota                  Subgroup_25
## 885 Bacteria                   Firmicutes                   Clostridia
## 886 Bacteria                 Bacteroidota                  Bacteroidia
## 887 Bacteria                 Bacteroidota                  Bacteroidia
## 888 Bacteria                   Firmicutes                   Clostridia
## 889 Bacteria                   Firmicutes                   Clostridia
## 890 Bacteria                   Firmicutes                   Clostridia
## 891 Bacteria                   Firmicutes           Desulfitobacteriia
## 892 Bacteria              Acidobacteriota             Vicinamibacteria
## 893 Bacteria              Acidobacteriota             Vicinamibacteria
## 894 Bacteria               Proteobacteria          Alphaproteobacteria
## 895 Bacteria               Proteobacteria          Gammaproteobacteria
## 896 Bacteria               Fusobacteriota                Fusobacteriia
## 897 Bacteria               Proteobacteria          Gammaproteobacteria
## 898 Bacteria               Proteobacteria          Alphaproteobacteria
## 899 Bacteria               Proteobacteria          Alphaproteobacteria
## 900 Bacteria                 Nitrospirota                       4-29-1
## 901 Bacteria               Proteobacteria          Gammaproteobacteria
## 902 Bacteria              Gemmatimonadota             Gemmatimonadetes
## 903 Bacteria             Actinobacteriota               Actinobacteria
## 904 Bacteria               Proteobacteria          Alphaproteobacteria
## 905 Bacteria                  Chloroflexi                 Chloroflexia
## 906 Bacteria                 Deinococcota                   Deinococci
## 907 Bacteria                   Firmicutes                   Clostridia
## 908 Bacteria               Proteobacteria          Gammaproteobacteria
## 909 Bacteria                   Firmicutes                      Bacilli
## 910 Bacteria            Verrucomicrobiota                Lentisphaeria
## 911 Bacteria            Verrucomicrobiota                Lentisphaeria
## 912 Bacteria                   Firmicutes                   Clostridia
## 913 Bacteria                 Bacteroidota                  Bacteroidia
## 914 Bacteria                 Bacteroidota                  Bacteroidia
## 915 Bacteria                 Bacteroidota                  Bacteroidia
## 916 Bacteria                 Bacteroidota                  Bacteroidia
## 917 Bacteria                 Bacteroidota                  Bacteroidia
## 918 Bacteria                 Bacteroidota                  Bacteroidia
## 919 Bacteria                   Firmicutes                   Clostridia
## 920 Bacteria                 Bacteroidota                  Bacteroidia
## 921 Bacteria                 Bacteroidota                  Bacteroidia
## 922 Bacteria                 Bacteroidota                  Bacteroidia
## 923 Bacteria                 Bacteroidota                  Bacteroidia
## 924 Bacteria                 Bacteroidota                  Bacteroidia
## 925 Bacteria                 Bacteroidota                  Bacteroidia
## 926 Bacteria                 Bacteroidota                  Bacteroidia
## 927 Bacteria                 Bacteroidota                  Bacteroidia
## 928 Bacteria                 Bacteroidota                  Bacteroidia
## 929 Bacteria                 Bacteroidota                  Bacteroidia
## 930 Bacteria               Proteobacteria          Gammaproteobacteria
## 931 Bacteria                   Firmicutes                   Clostridia
## 932 Bacteria                   Firmicutes                      Bacilli
## 933 Bacteria             Bdellovibrionota                  Oligoflexia
## 934 Bacteria               Proteobacteria          Alphaproteobacteria
## 935 Bacteria                   Firmicutes                   Clostridia
## 936 Bacteria               Proteobacteria          Alphaproteobacteria
## 937 Bacteria                 Bacteroidota                  Bacteroidia
## 938 Bacteria                   Firmicutes                   Clostridia
## 939 Bacteria                 Bacteroidota                  Bacteroidia
## 940 Bacteria                 Bacteroidota                  Bacteroidia
## 941 Bacteria               Proteobacteria          Gammaproteobacteria
## 942 Bacteria                   Firmicutes                   Clostridia
## 943 Bacteria                   Firmicutes                Negativicutes
## 944 Bacteria                   Firmicutes                      Bacilli
## 945 Bacteria                   Firmicutes                Negativicutes
## 946 Bacteria                   Firmicutes                Negativicutes
## 947 Bacteria                   Firmicutes                Negativicutes
## 948 Bacteria                   Firmicutes                      Bacilli
## 949 Bacteria                   Firmicutes                   Clostridia
## 950 Bacteria                   Firmicutes                   Clostridia
## 951 Bacteria                   Firmicutes                   Clostridia
## 952 Bacteria                 Bacteroidota                  Bacteroidia
## 953 Bacteria                  Chloroflexi                       KD4-96
## 954 Bacteria              Elusimicrobiota                Elusimicrobia
## 955 Bacteria                Cyanobacteria             Vampirivibrionia
## 956 Bacteria                Cyanobacteria               Cyanobacteriia
## 957 Bacteria                   Firmicutes                   Clostridia
## 958 Bacteria                   Firmicutes                   Clostridia
## 959 Bacteria            Verrucomicrobiota             Verrucomicrobiae
## 960 Bacteria            Verrucomicrobiota             Verrucomicrobiae
##                                   Order                                Family
## 1                            uncultured                            uncultured
## 2                          Chromatiales                         Chromatiaceae
## 3                    Syntrophomonadales                   Syntrophomonadaceae
## 4                     Symbiobacteriales                     Symbiobacteraceae
## 5                      Gemmatimonadales                     Gemmatimonadaceae
## 6                      Gemmatimonadales                     Gemmatimonadaceae
## 7                        Chloroflexales                    Herpetosiphonaceae
## 8                          ADurb.Bin180                          ADurb.Bin180
## 9                        Chloroflexales                        Roseiflexaceae
## 10                       Chloroflexales                        Roseiflexaceae
## 11                       Chloroflexales                        Roseiflexaceae
## 12                          RBG-13-54-9                           RBG-13-54-9
## 13                       Anaerolineales                       Anaerolineaceae
## 14                       Anaerolineales                       Anaerolineaceae
## 15                              SBR1031                               SBR1031
## 16                              SBR1031                                   A4b
## 17                    Thermomicrobiales                          JG30-KF-CM45
## 18                                 S085                                  S085
## 19                         JG30-KF-CM66                          JG30-KF-CM66
## 20                        Caldilineales                        Caldilineaceae
## 21                        Caldilineales                        Caldilineaceae
## 22                                 1-20                                  1-20
## 23                    Ktedonobacterales                           JG30-KF-AS9
## 24                       Chloroflexales                       Chloroflexaceae
## 25                       Chloroflexales                       Chloroflexaceae
## 26                       Chloroflexales                       Chloroflexaceae
## 27                       Chloroflexales                       Chloroflexaceae
## 28                               SHA-26                                SHA-26
## 29                     Chthonomonadales                     Chthonomonadaceae
## 30                            Elev-1554                             Elev-1554
## 31                          Gitt-GS-136                           Gitt-GS-136
## 32                        Kallotenuales                               AKIW781
## 33                                OLB14                                 OLB14
## 34                    Ktedonobacterales                    Ktedonobacteraceae
## 35                    Ktedonobacterales                    Ktedonobacteraceae
## 36                    Ktedonobacterales                    Ktedonobacteraceae
## 37                    Ktedonobacterales                    Ktedonobacteraceae
## 38                    Ktedonobacterales                    Ktedonobacteraceae
## 39                    Ktedonobacterales                    Ktedonobacteraceae
## 40                    Ktedonobacterales                    Ktedonobacteraceae
## 41                    Ktedonobacterales                    Ktedonobacteraceae
## 42                    Ktedonobacterales                    Ktedonobacteraceae
## 43                      Rokubacteriales                       Rokubacteriales
## 44                            B12-WMSP1                             B12-WMSP1
## 45                                GAL15                                 GAL15
## 46                                 TK10                                  TK10
## 47                                WPS-2                                 WPS-2
## 48                       Brevinematales                       Brevinemataceae
## 49          Absconditabacteriales_(SR1)           Absconditabacteriales_(SR1)
## 50                     Ardenticatenales                     Ardenticatenaceae
## 51                         Elev-16S-573                          Elev-16S-573
## 52                     Blastocatellales                     Blastocatellaceae
## 53                      Pyrinomonadales                      Pyrinomonadaceae
## 54                     Blastocatellales                     Blastocatellaceae
## 55                          Lineage_IIc                           Lineage_IIc
## 56                          Lineage_IIa                           Lineage_IIa
## 57                          Lineage_IIb                           Lineage_IIb
## 58                      Xanthomonadales                      Xanthomonadaceae
## 59                       Acidobacteriae                        Acidobacteriae
## 60                       Solibacterales                       Solibacteraceae
## 61                               GOUTB8                                GOUTB8
## 62                        Nitrospirales                        Nitrospiraceae
## 63                         Myxococcales                         Myxococcaceae
## 64                     Acidobacteriales        Acidobacteriaceae_(Subgroup_1)
## 65                     Acidobacteriales        Acidobacteriaceae_(Subgroup_1)
## 66                     Acidobacteriales        Acidobacteriaceae_(Subgroup_1)
## 67                     Acidobacteriales        Acidobacteriaceae_(Subgroup_1)
## 68                     Acidobacteriales                            uncultured
## 69                         Sumerlaeales                         Sumerlaeaceae
## 70                     Longimicrobiales                     Longimicrobiaceae
## 71                     Longimicrobiales                     Longimicrobiaceae
## 72                          Subgroup_17                           Subgroup_17
## 73                               MVP-15                                MVP-15
## 74                          bacteriap25                           bacteriap25
## 75         SAR324_clade(Marine_group_B)          SAR324_clade(Marine_group_B)
## 76                       Spirochaetales                       Spirochaetaceae
## 77                       Spirochaetales                       Spirochaetaceae
## 78                       Spirochaetales                       Spirochaetaceae
## 79                           uncultured                            uncultured
## 80                        Leptospirales                        Leptospiraceae
## 81                        Leptospirales                        Leptospiraceae
## 82                           Gaiellales                            uncultured
## 83                  Solirubrobacterales                  Solirubrobacteraceae
## 84                       Microtrichales                       Microtrichaceae
## 85                       Microtrichales                    Ilumatobacteraceae
## 86                       Microtrichales                    Ilumatobacteraceae
## 87                       Microtrichales                    Ilumatobacteraceae
## 88                           Euzebyales                           Euzebyaceae
## 89                       Microtrichales                             Iamiaceae
## 90                       Microtrichales                    Ilumatobacteraceae
## 91                            IMCC26256                             IMCC26256
## 92                       Microtrichales                            uncultured
## 93                       Microtrichales                        Microtrichales
## 94                       Microtrichales                       Microtrichaceae
## 95                  Propionibacteriales                  Propionibacteriaceae
## 96                  Propionibacteriales                  Propionibacteriaceae
## 97                  Propionibacteriales                  Propionibacteriaceae
## 98                           Euzebyales                           Euzebyaceae
## 99                        Micrococcales                      Dermatophilaceae
## 100                    Cyanobacteriales                           Nostocaceae
## 101                 Streptosporangiales                   Thermomonosporaceae
## 102                          Gaiellales                           Gaiellaceae
## 103                 Solirubrobacterales                  Solirubrobacteraceae
## 104                 Solirubrobacterales                  Solirubrobacteraceae
## 105                 Solirubrobacterales                  Solirubrobacteraceae
## 106                  Chthoniobacterales                     Terrimicrobiaceae
## 107                 Solirubrobacterales                  Solirubrobacteraceae
## 108                 Solirubrobacterales                  Solirubrobacteraceae
## 109                          uncultured                            uncultured
## 110                      Bryobacterales                       Bryobacteraceae
## 111                 Solirubrobacterales                                 67-14
## 112                           vadinHA49                             vadinHA49
## 113                          uncultured                            uncultured
## 114                        Kryptoniales                                 BSV26
## 115                        Zixibacteria                          Zixibacteria
## 116                     Fibrobacterales                                  B122
## 117                     Fibrobacterales                       Fibrobacterales
## 118                   Ignavibacteriales                            SR-FBR-L83
## 119                   Ignavibacteriales                             PHOS-HE36
## 120                     Kapabacteriales                       Kapabacteriales
## 121                              SJA-28                                SJA-28
## 122                      Rhodothermales                       Rhodothermaceae
## 123                      Rhodothermales                       Rhodothermaceae
## 124                    Chthonomonadales                      Chthonomonadales
## 125                     Armatimonadales                       Armatimonadales
## 126                     Armatimonadales                      Armatimonadaceae
## 127                     Micavibrionales                            uncultured
## 128                         Rhizobiales                     Xanthobacteraceae
## 129                       Micropepsales                        Micropepsaceae
## 130                         Rhizobiales                            KF-JG30-B3
## 131                         Rhizobiales                                 A0839
## 132                          Elsterales                            uncultured
## 133                          uncultured                            uncultured
## 134                         Rhizobiales                            uncultured
## 135                        Tistrellales                       Geminicoccaceae
## 136                        Tistrellales                       Geminicoccaceae
## 137                        Tistrellales                       Geminicoccaceae
## 138                           Dongiales                            Dongiaceae
## 139                    Defluviicoccales                     Defluviicoccaceae
## 140                      Azospirillales                       Azospirillaceae
## 141                     Acetobacterales                      Acetobacteraceae
## 142                     Acetobacterales                      Acetobacteraceae
## 143                     Acetobacterales                      Acetobacteraceae
## 144                  Steroidobacterales                   Steroidobacteraceae
## 145                     Acetobacterales                      Acetobacteraceae
## 146                     Acetobacterales                      Acetobacteraceae
## 147                     Acetobacterales                      Acetobacteraceae
## 148                     Acetobacterales                      Acetobacteraceae
## 149                     Acetobacterales                      Acetobacteraceae
## 150                     Acetobacterales                      Acetobacteraceae
## 151                       Reyranellales                        Reyranellaceae
## 152                      Azospirillales                            uncultured
## 153                    Rhodospirillales                     Rhodospirillaceae
## 154                       Rickettsiales                           Fokiniaceae
## 155                       Rickettsiales                         Rickettsiales
## 156                       Rickettsiales                                SM2D12
## 157                     Micavibrionales                      Micavibrionaceae
## 158                    Sphingomonadales                     Sphingomonadaceae
## 159                    Sphingomonadales                     Sphingomonadaceae
## 160                    Sphingomonadales                     Sphingomonadaceae
## 161                    Sphingomonadales                     Sphingomonadaceae
## 162                    Rhodospirillales                   Magnetospirillaceae
## 163                    Sphingomonadales                     Sphingomonadaceae
## 164                    Sphingomonadales                     Sphingomonadaceae
## 165                    Sphingomonadales                     Sphingomonadaceae
## 166                    Sphingomonadales                     Sphingomonadaceae
## 167                         Rhizobiales                      Beijerinckiaceae
## 168                         Rhizobiales                             Labraceae
## 169                         Rhizobiales                      Beijerinckiaceae
## 170                         Rhizobiales            Rhizobiales_Incertae_Sedis
## 171                         Rhizobiales                     Xanthobacteraceae
## 172                         Rhizobiales                    Methyloligellaceae
## 173                         Rhizobiales                      Beijerinckiaceae
## 174                         Rhizobiales                     Xanthobacteraceae
## 175                         Rhizobiales                    Methyloligellaceae
## 176                         Rhizobiales            Rhizobiales_Incertae_Sedis
## 177                     Rhodobacterales                      Rhodobacteraceae
## 178                         Rhizobiales                      Beijerinckiaceae
## 179                         Rhizobiales                      Beijerinckiaceae
## 180                     Caulobacterales                      Caulobacteraceae
## 181                     Caulobacterales                      Caulobacteraceae
## 182                     Caulobacterales                      Caulobacteraceae
## 183                         Rhizobiales                          Rhizobiaceae
## 184                         Rhizobiales                     Xanthobacteraceae
## 185                         Rhizobiales                     Xanthobacteraceae
## 186                         Rhizobiales                     Xanthobacteraceae
## 187                         Rhizobiales                      Beijerinckiaceae
## 188                         Rhizobiales                           Devosiaceae
## 189                         Rhizobiales            Rhizobiales_Incertae_Sedis
## 190                         Rhizobiales            Rhizobiales_Incertae_Sedis
## 191                         Rhizobiales                     Rhodomicrobiaceae
## 192                         Rhizobiales                     Hyphomicrobiaceae
## 193                         Rhizobiales                     Hyphomicrobiaceae
## 194                         Rhizobiales                           Kaistiaceae
## 195                         Rhizobiales                     Hyphomicrobiaceae
## 196                         Rhizobiales                     Xanthobacteraceae
## 197                         Rhizobiales                      Beijerinckiaceae
## 198                         Rhizobiales                      Beijerinckiaceae
## 199                     Caulobacterales                       Hyphomonadaceae
## 200                         Rhizobiales                                 D05-2
## 201                     Caulobacterales                      Caulobacteraceae
## 202                 Paracaedibacterales                  Paracaedibacteraceae
## 203                 Paracaedibacterales                  Paracaedibacteraceae
## 204                        Holosporales                         Holosporaceae
## 205                       Rickettsiales                                   AB1
## 206                        Holosporales                         Holosporaceae
## 207                   Sericytochromatia                     Sericytochromatia
## 208  Alphaproteobacteria_Incertae_Sedis                        Unknown_Family
## 209                   Cardiobacteriales                 Wohlfahrtiimonadaceae
## 210                   Cardiobacteriales                 Wohlfahrtiimonadaceae
## 211                  Diplorickettsiales                   Diplorickettsiaceae
## 212                   Competibacterales                    Competibacteraceae
## 213                     Fibrobacterales                      Fibrobacteraceae
## 214                               HOC36                                 HOC36
## 215                Acidiferrobacterales                 Acidiferrobacteraceae
## 216                     Xanthomonadales                    Rhodanobacteraceae
## 217                     Pseudomonadales                         Moraxellaceae
## 218                               CCD24                                 CCD24
## 219                              PLTA13                                PLTA13
## 220                    Salinisphaerales                        Solimonadaceae
## 221                    Salinisphaerales                        Solimonadaceae
## 222                        JG36-TzT-191                          JG36-TzT-191
## 223                         Coxiellales                          Coxiellaceae
## 224                          JG36-GS-52                            JG36-GS-52
## 225                  Steroidobacterales                   Steroidobacteraceae
## 226                   Bdellovibrionales                    Bdellovibrionaceae
## 227                  Diplorickettsiales                   Diplorickettsiaceae
## 228                   Oceanospirillales                        Halomonadaceae
## 229                   Oceanospirillales                        Halomonadaceae
## 230                     Nitrosococcales                      Nitrosococcaceae
## 231                         KF-JG30-C25                           KF-JG30-C25
## 232                       Thiotrichales                        Thiotrichaceae
## 233               Thermoanaerobaculales                Thermoanaerobaculaceae
## 234                     Methylococcales                      Methylococcaceae
## 235                          CHAB-XI-27                            CHAB-XI-27
## 236                 Gammaproteobacteria                   Gammaproteobacteria
## 237  Gammaproteobacteria_Incertae_Sedis                        Unknown_Family
## 238  Gammaproteobacteria_Incertae_Sedis                        Unknown_Family
## 239  Gammaproteobacteria_Incertae_Sedis                        Unknown_Family
## 240                        F9P41300-M23                          F9P41300-M23
## 241                          uncultured                            uncultured
## 242                     Methylococcales                     Methylomonadaceae
## 243                              MBNT15                                MBNT15
## 244                             RCP2-54                               RCP2-54
## 245                       Deinococcales                          Trueperaceae
## 246                       Deinococcales                        Deinococcaceae
## 247                       Deinococcales                        Deinococcaceae
## 248                  Syntrophorhabdales                   Syntrophorhabdaceae
## 249                   Bdellovibrionales                    Bdellovibrionaceae
## 250                        Syntrophales                         Syntrophaceae
## 251                        Syntrophales                            uncultured
## 252                   Desulfobacterales                    Desulfosarcinaceae
## 253                       Geobacterales                        Geobacteraceae
## 254                       Geobacterales                        Geobacteraceae
## 255                     Desulfobaccales                      Desulfobaccaceae
## 256                    Silvanigrellales                     Silvanigrellaceae
## 257                     Rokubacteriales                                  WX65
## 258                              BD7-11                                BD7-11
## 259                       Nannocystales                        Nannocystaceae
## 260                       Nannocystales                        Nannocystaceae
## 261                      Bradymonadales                        Bradymonadales
## 262                       Oligoflexales                            uncultured
## 263                       Oligoflexales                         Oligoflexales
## 264                                PB19                                  PB19
## 265                             Blfdi19                               Blfdi19
## 266                               NKB15                                 NKB15
## 267                     Acetobacterales                      Acetobacteraceae
## 268                        Myxococcales                         Myxococcaceae
## 269                        Myxococcales                    Vulgatibacteraceae
## 270                        Myxococcales                         Myxococcaceae
## 271                        Polyangiales                    Phaselicystidaceae
## 272                            MSB-4B10                              MSB-4B10
## 273                        Haliangiales                         Haliangiaceae
## 274                           VHS-B4-70                             VHS-B4-70
## 275                             mle1-27                               mle1-27
## 276                        Polyangiales                         Polyangiaceae
## 277                        Polyangiales                         Polyangiaceae
## 278                        Polyangiales                         Polyangiaceae
## 279                        Polyangiales                         Polyangiaceae
## 280                        Polyangiales                         Polyangiaceae
## 281                        Polyangiales                            uncultured
## 282                        Polyangiales                               BIrii41
## 283                               NB1-j                                 NB1-j
## 284                           FFCH16767                             FFCH16767
## 285                        Polyangiales                       Sandaracinaceae
## 286                    Acidobacteriales                       Koribacteraceae
## 287                          Subgroup_2                            Subgroup_2
## 288                         Subgroup_11                           Subgroup_11
## 289                    Latescibacterota                      Latescibacterota
## 290                               11-24                                 11-24
## 291                    Blastocatellales                     Blastocatellaceae
## 292                    Blastocatellales                     Blastocatellaceae
## 293                     Gracilibacteria                       Gracilibacteria
## 294                     JGI_0000069-P22                       JGI_0000069-P22
## 295                  Methanomicrobiales                     Methanoregulaceae
## 296                  Methanobacteriales                   Methanobacteriaceae
## 297                  Methanobacteriales                   Methanobacteriaceae
## 298        Candidatus_Magasanikbacteria          Candidatus_Magasanikbacteria
## 299         Candidatus_Yanofskybacteria           Candidatus_Yanofskybacteria
## 300                       Parcubacteria                         Parcubacteria
## 301                   Saccharimonadales                     Saccharimonadales
## 302                    Fimbriimonadales                     Fimbriimonadaceae
## 303                    Fimbriimonadales                     Fimbriimonadaceae
## 304                   Saccharimonadales                                  LWQ8
## 305                   Saccharimonadales                                 WWH38
## 306                   Saccharimonadales                                   S32
## 307                   Saccharimonadales                    Saccharimonadaceae
## 308                     Pseudomonadales                      Pseudomonadaceae
## 309                   Campylobacterales                     Helicobacteraceae
## 310                       Omnitrophales                        Omnitrophaceae
## 311                  Vicinamibacterales                            uncultured
## 312                   Campylobacterales                    Campylobacteraceae
## 313                   Campylobacterales                            uncultured
## 314                   Campylobacterales                     Sulfurimonadaceae
## 315                   Campylobacterales                       Arcobacteraceae
## 316                   Nitrososphaerales                    Nitrososphaeraceae
## 317                     Nitrosotaleales                      Nitrosotaleaceae
## 318                  Methanosarciniales                      Methanosaetaceae
## 319                      Halobacterales                      Halomicrobiaceae
## 320                  Methanomicrobiales                 Methanocorpusculaceae
## 321             Methanomassiliicoccales               Methanomethylophilaceae
## 322                     Woesearchaeales                       Woesearchaeales
## 323                          Babeliales                              UBA12409
## 324                          Babeliales                           Babeliaceae
## 325                          Babeliales                        Vermiphilaceae
## 326                          Babeliales                            Babeliales
## 327                        Holophagales                         Holophagaceae
## 328                          Opitutales                           Opitutaceae
## 329                          Opitutales                           Opitutaceae
## 330                          Opitutales                      Puniceicoccaceae
## 331                          uncultured                            uncultured
## 332                          Opitutales                      Puniceicoccaceae
## 333                          Opitutales                           Opitutaceae
## 334                      Pedosphaerales                       Pedosphaeraceae
## 335                      Pedosphaerales                       Pedosphaeraceae
## 336                  Chthoniobacterales                Xiphinematobacteraceae
## 337                  Chthoniobacterales                   Chthoniobacteraceae
## 338                  Verrucomicrobiales                   Verrucomicrobiaceae
## 339                 Methylacidiphilales                  Methylacidiphilaceae
## 340                  Verrucomicrobiales                        Rubritaleaceae
## 341                      Pedosphaerales                       Pedosphaeraceae
## 342                      Pedosphaerales                       Pedosphaeraceae
## 343                      Pedosphaerales                       Pedosphaeraceae
## 344                      Pedosphaerales                       Pedosphaeraceae
## 345                      Pedosphaerales                       Pedosphaeraceae
## 346                 S-BQ2-57_soil_group                   S-BQ2-57_soil_group
## 347                  Chthoniobacterales                               01D2Z36
## 348                  Chthoniobacterales                   Chthoniobacteraceae
## 349                                UA11                                  UA11
## 350                  Verrucomicrobiales                   Verrucomicrobiaceae
## 351                  Verrucomicrobiales                   Verrucomicrobiaceae
## 352                  Verrucomicrobiales                   Verrucomicrobiaceae
## 353                    Leptospirillales                     Leptospirillaceae
## 354                         Subgroup_22                           Subgroup_22
## 355                  Vicinamibacterales                   Vicinamibacteraceae
## 356                         Subgroup_18                           Subgroup_18
## 357                            AT-s3-28                              AT-s3-28
## 358                          Subgroup_7                            Subgroup_7
## 359                    Entotheonellales                     Entotheonellaceae
## 360                        Myxococcales                         Myxococcaceae
## 361                  Hydrogenedentiales                    Hydrogenedensaceae
## 362                        Pla4_lineage                          Pla4_lineage
## 363               Thermoanaerobaculales                Thermoanaerobaculaceae
## 364                             LD1-PB3                               LD1-PB3
## 365                            WCHB1-41                              WCHB1-41
## 366                              DS-100                                DS-100
## 367                  Alicyclobacillales                   Alicyclobacillaceae
## 368                        Chlamydiales                     Parachlamydiaceae
## 369                        Chlamydiales                     Parachlamydiaceae
## 370                        Chlamydiales                         Chlamydiaceae
## 371                        Chlamydiales                     Parachlamydiaceae
## 372                        Chlamydiales                     Parachlamydiaceae
## 373                        Chlamydiales                         Chlamydiaceae
## 374                        Chlamydiales                          Simkaniaceae
## 375                        Chlamydiales                          Simkaniaceae
## 376                        Chlamydiales                     Parachlamydiaceae
## 377                        Chlamydiales                       Criblamydiaceae
## 378                        Chlamydiales                          Simkaniaceae
## 379 Peptostreptococcales-Tissierellales   Peptostreptococcales-Tissierellales
## 380 Peptostreptococcales-Tissierellales   Peptostreptococcales-Tissierellales
## 381 Peptostreptococcales-Tissierellales                       Fusibacteraceae
## 382 Peptostreptococcales-Tissierellales   Peptostreptococcales-Tissierellales
## 383 Peptostreptococcales-Tissierellales   Peptostreptococcales-Tissierellales
## 384                       Synergistales                        Synergistaceae
## 385                       Synergistales                        Synergistaceae
## 386                  Christensenellales                   Christensenellaceae
## 387                               OPB41                                 OPB41
## 388                    Coriobacteriales                            uncultured
## 389                    Coriobacteriales                          Atopobiaceae
## 390                    Coriobacteriales                          Atopobiaceae
## 391                     Xanthomonadales                      Xanthomonadaceae
## 392                     Xanthomonadales                    Rhodanobacteraceae
## 393                     Xanthomonadales                    Rhodanobacteraceae
## 394                       Legionellales                        Legionellaceae
## 395                     Cellvibrionales                        Porticoccaceae
## 396                         KI89A_clade                           KI89A_clade
## 397                     Burkholderiales                     Nitrosomonadaceae
## 398                             FCPU426                               FCPU426
## 399                               WD260                                 WD260
## 400               Thermoactinomycetales                Thermoactinomycetaceae
## 401               Thermoactinomycetales                Thermoactinomycetaceae
## 402               Thermoactinomycetales                Thermoactinomycetaceae
## 403                  Methylomirabilales                   Methylomirabilaceae
## 404                  Methylomirabilales                   Methylomirabilaceae
## 405                                 AD3                                   AD3
## 406                     Dadabacteriales                       Dadabacteriales
## 407                       Bacteroidales                       Barnesiellaceae
## 408                       Bacteroidales                     Paludibacteraceae
## 409                       Bacteroidales                                  F082
## 410                       Bacteroidales                              p-251-o5
## 411                       Bacteroidales                         Rikenellaceae
## 412                     Oscillospirales                      Oscillospiraceae
## 413                    Flavobacteriales                     Flavobacteriaceae
## 414                     Chitinophagales                      Chitinophagaceae
## 415                       Bacteroidales                         Rikenellaceae
## 416                  Sphingobacteriales                               AKYH767
## 417                       Bacteroidales               Bacteroidetes_vadinHA17
## 418                        Cytophagales                     Hymenobacteraceae
## 419                        Cytophagales                         Cytophagaceae
## 420                        Cytophagales                         Cytophagaceae
## 421                        Cytophagales                     Cyclobacteriaceae
## 422                        Cytophagales                     Hymenobacteraceae
## 423                        Cytophagales                       Microscillaceae
## 424                        Cytophagales                         Cytophagaceae
## 425                     Chitinophagales                      Chitinophagaceae
## 426                        Cytophagales                       Microscillaceae
## 427                        Cytophagales                       Microscillaceae
## 428                        Cytophagales                         Spirosomaceae
## 429                        Cytophagales                         Spirosomaceae
## 430                        Cytophagales                         Spirosomaceae
## 431                       Bacteroidales                  Williamwhitmaniaceae
## 432                       Bacteroidales                   Bacteroidetes_BD2-2
## 433                  Sphingobacteriales                              ST-12K33
## 434                       Bacteroidales                     Marinilabiliaceae
## 435                       Bacteroidales                     Paludibacteraceae
## 436                       Bacteroidales                     Paludibacteraceae
## 437                       Bacteroidales                     Paludibacteraceae
## 438                       Bacteroidales                         Bacteroidales
## 439                       Bacteroidales                        Muribaculaceae
## 440                       Bacteroidales                       Barnesiellaceae
## 441                     Chitinophagales                            uncultured
## 442                       Bacteroidales                    Prolixibacteraceae
## 443                  Sphingobacteriales                   Sphingobacteriaceae
## 444                  Sphingobacteriales                                Z4MB62
## 445                       Bacteroidales                             vadinHA21
## 446                  Sphingobacteriales                            env.OPS_17
## 447                    Flavobacteriales                      NS9_marine_group
## 448                  Sphingobacteriales                  NS11-12_marine_group
## 449                     Chitinophagales                        Saprospiraceae
## 450                     Chitinophagales                        Saprospiraceae
## 451                     Chitinophagales                      Chitinophagaceae
## 452                     Chitinophagales                      Chitinophagaceae
## 453                     Chitinophagales                      Chitinophagaceae
## 454                     Chitinophagales                      Chitinophagaceae
## 455                     Chitinophagales                      Chitinophagaceae
## 456                     Chitinophagales                      Chitinophagaceae
## 457                     Chitinophagales                      Chitinophagaceae
## 458                     Chitinophagales                      Chitinophagaceae
## 459                     Chitinophagales                      Chitinophagaceae
## 460                     Chitinophagales                      Chitinophagaceae
## 461                     Chitinophagales                      Chitinophagaceae
## 462                     Chitinophagales                      Chitinophagaceae
## 463                     Chitinophagales                      Chitinophagaceae
## 464                     Chitinophagales                      Chitinophagaceae
## 465                  Sphingobacteriales                   Sphingobacteriaceae
## 466                    Flavobacteriales                     Crocinitomicaceae
## 467                       Bacteroidales                  Williamwhitmaniaceae
## 468                       Bacteroidales                                CR-115
## 469                       Bacteroidales                         Rikenellaceae
## 470                       Bacteroidales                                 PeH15
## 471                       Bacteroidales                M2PB4-65_termite_group
## 472                       Bacteroidales                COB_P4-1_termite_group
## 473                    Flavobacteriales                     Crocinitomicaceae
## 474                  Sphingobacteriales                     Lentimicrobiaceae
## 475                  Sphingobacteriales                     Lentimicrobiaceae
## 476                       Bacteroidales                    Prolixibacteraceae
## 477                       Bacteroidales                                  SB-5
## 478                    Flavobacteriales                         Weeksellaceae
## 479                    Flavobacteriales                         Weeksellaceae
## 480                    Flavobacteriales                         Weeksellaceae
## 481                    Flavobacteriales                         Weeksellaceae
## 482                    Flavobacteriales                     Flavobacteriaceae
## 483                        Cytophagales                         Spirosomaceae
## 484                        Cytophagales                         Spirosomaceae
## 485                        Cytophagales                         Spirosomaceae
## 486                        Cytophagales                         Cytophagaceae
## 487                        Cytophagales                         Spirosomaceae
## 488                        Cytophagales                         Spirosomaceae
## 489                        Cytophagales                       Microscillaceae
## 490                        Cytophagales                       Microscillaceae
## 491                  Sphingobacteriales                   Sphingobacteriaceae
## 492                  Sphingobacteriales                   Sphingobacteriaceae
## 493                  Sphingobacteriales                   Sphingobacteriaceae
## 494                  Sphingobacteriales                   Sphingobacteriaceae
## 495                    Flavobacteriales                     Flavobacteriaceae
## 496                    Flavobacteriales                     Flavobacteriaceae
## 497                          Gemmatales                           Gemmataceae
## 498                          Gemmatales                           Gemmataceae
## 499                          Gemmatales                           Gemmataceae
## 500                    Planctomycetales                       Schlesneriaceae
## 501                       Isosphaerales                        Isosphaeraceae
## 502                    Planctomycetales                     Rubinisphaeraceae
## 503                    Planctomycetales                            uncultured
## 504                    Planctomycetales                           Gimesiaceae
## 505                    Planctomycetales                     Rubinisphaeraceae
## 506                    Planctomycetales                       Schlesneriaceae
## 507                        Pirellulales                         Pirellulaceae
## 508                        Pirellulales                         Pirellulaceae
## 509                        Pirellulales                         Pirellulaceae
## 510                        Pirellulales                         Pirellulaceae
## 511                        Pirellulales                         Pirellulaceae
## 512                        Pirellulales                         Pirellulaceae
## 513                        Pirellulales                         Pirellulaceae
## 514                        Pirellulales                         Pirellulaceae
## 515                        Pirellulales                         Pirellulaceae
## 516                               OM190                                 OM190
## 517                                S-70                                  S-70
## 518                        Pla1_lineage                          Pla1_lineage
## 519                     Phycisphaerales                      Phycisphaeraceae
## 520                     Phycisphaerales                      Phycisphaeraceae
## 521                     Phycisphaerales                      Phycisphaeraceae
## 522                               MSBL9                                 SG8-4
## 523                     Phycisphaerales               AKAU3564_sediment_group
## 524                    Tepidisphaerales                     WD2101_soil_group
## 525                    Tepidisphaerales                     Tepidisphaeraceae
## 526                    Tepidisphaerales                     Tepidisphaeraceae
## 527                          Gemmatales                           Gemmataceae
## 528                          Gemmatales                           Gemmataceae
## 529                          uncultured                            uncultured
## 530                          Gemmatales                           Gemmataceae
## 531                          Gemmatales                           Gemmataceae
## 532                       Isosphaerales                        Isosphaeraceae
## 533                       Isosphaerales                        Isosphaeraceae
## 534                       Isosphaerales                        Isosphaeraceae
## 535                       Isosphaerales                        Isosphaeraceae
## 536                       Isosphaerales                        Isosphaeraceae
## 537                       Isosphaerales                        Isosphaeraceae
## 538           Candidatus_Nomurabacteria             Candidatus_Nomurabacteria
## 539                  Desulfovibrionales                   Desulfovibrionaceae
## 540                  Desulfovibrionales                   Desulfovibrionaceae
## 541                          Subgroup_5                            Subgroup_5
## 542                     Oscillospirales                       Oscillospirales
## 543                     Oscillospirales                       Ruminococcaceae
## 544                     Oscillospirales                       Ruminococcaceae
## 545                     Oscillospirales                       Ruminococcaceae
## 546                     Lactobacillales                      Streptococcaceae
## 547                     Lactobacillales                      Streptococcaceae
## 548                     Rubrobacterales                     Rubrobacteriaceae
## 549                           MB-A2-108                             MB-A2-108
## 550                   Micromonosporales                    Micromonosporaceae
## 551                           FFCH16263                             FFCH16263
## 552                 Propionibacteriales                       Nocardioidaceae
## 553                               PeM15                                 PeM15
## 554                   Pseudonocardiales                    Pseudonocardiaceae
## 555                   Micromonosporales                    Micromonosporaceae
## 556                   Micromonosporales                    Micromonosporaceae
## 557                   Micromonosporales                    Micromonosporaceae
## 558                          Frankiales                           Frankiaceae
## 559                   Corynebacteriales      Corynebacteriales_Incertae_Sedis
## 560                   Corynebacteriales                    Corynebacteriaceae
## 561                   Corynebacteriales                    Corynebacteriaceae
## 562                   Corynebacteriales                          Nocardiaceae
## 563                   Corynebacteriales                            uncultured
## 564                   Corynebacteriales                          Nocardiaceae
## 565                   Corynebacteriales                     Corynebacteriales
## 566                          Frankiales                   Cryptosporangiaceae
## 567                          Frankiales                   Geodermatophilaceae
## 568                      Kineosporiales                       Kineosporiaceae
## 569                      Kineosporiales                       Kineosporiaceae
## 570                     Actinomycetales                      Actinomycetaceae
## 571                       Micrococcales                        Micrococcaceae
## 572                       Micrococcales                    Intrasporangiaceae
## 573                       Micrococcales                      Dermatophilaceae
## 574                       Micrococcales                      Dermatophilaceae
## 575                       Micrococcales                    Intrasporangiaceae
## 576                       Micrococcales                      Beutenbergiaceae
## 577                       Micrococcales                      Dermabacteraceae
## 578                       Micrococcales                     Microbacteriaceae
## 579                      Kineosporiales                       Kineosporiaceae
## 580                     Actinomycetales        Actinomycetales_Incertae_Sedis
## 581                      Kineosporiales                       Kineosporiaceae
## 582                      Kineosporiales                       Kineosporiaceae
## 583                    Catenulisporales                     Catenulisporaceae
## 584                 Propionibacteriales                  Propionibacteriaceae
## 585                   Pseudonocardiales                    Pseudonocardiaceae
## 586                    Streptomycetales                     Streptomycetaceae
## 587                   Pseudonocardiales                    Pseudonocardiaceae
## 588                   Pseudonocardiales                    Pseudonocardiaceae
## 589                   Micromonosporales                    Micromonosporaceae
## 590                   Micromonosporales                    Micromonosporaceae
## 591                   Micromonosporales                    Micromonosporaceae
## 592                   Micromonosporales                    Micromonosporaceae
## 593                          Frankiales                       Sporichthyaceae
## 594                    Streptomycetales                     Streptomycetaceae
## 595                          Frankiales                       Sporichthyaceae
## 596                   Pseudonocardiales                    Pseudonocardiaceae
## 597                          Frankiales                       Nakamurellaceae
## 598                 Propionibacteriales                       Nocardioidaceae
## 599                 Streptosporangiales                  Streptosporangiaceae
## 600                 Streptosporangiales                  Streptosporangiaceae
## 601                   Micromonosporales                    Micromonosporaceae
## 602                          Frankiales                       Acidothermaceae
## 603                 Streptosporangiales                   Thermomonosporaceae
## 604                          Frankiales                            Frankiales
## 605                 Streptosporangiales                   Thermomonosporaceae
## 606                 Propionibacteriales                  Propionibacteriaceae
## 607                 Propionibacteriales                  Propionibacteriaceae
## 608                 Propionibacteriales                       Nocardioidaceae
## 609                   Oceanospirillales                      Marinomonadaceae
## 610                     Burkholderiales                      Burkholderiaceae
## 611                     Burkholderiales                      Burkholderiaceae
## 612                     Burkholderiales                     Nitrosomonadaceae
## 613                        Holophagales                         Holophagaceae
## 614                     Burkholderiales                        Comamonadaceae
## 615                     Burkholderiales                        Comamonadaceae
## 616                     Burkholderiales                        Sutterellaceae
## 617                     Burkholderiales                      Oxalobacteraceae
## 618                     Burkholderiales                    Chromobacteriaceae
## 619                     Burkholderiales                         Neisseriaceae
## 620                     Burkholderiales                    Chromobacteriaceae
## 621                     Burkholderiales                    Chromobacteriaceae
## 622                     Burkholderiales                    Hydrogenophilaceae
## 623                     Burkholderiales                               SC-I-84
## 624                     Burkholderiales                                  A21b
## 625                     Burkholderiales                         Neisseriaceae
## 626                     Burkholderiales                         Neisseriaceae
## 627                     Burkholderiales                               TRA3-20
## 628                     Burkholderiales                                B1-7BS
## 629                     Lactobacillales                      Lactobacillaceae
## 630                       Aeromonadales                        Aeromonadaceae
## 631                    Rhodospirillales                            uncultured
## 632                      Lachnospirales                       Lachnospiraceae
## 633                      Lachnospirales                       Lachnospiraceae
## 634                     Oscillospirales                      Oscillospiraceae
## 635                     Oscillospirales                               UCG-010
## 636                     Oscillospirales                      Oscillospiraceae
## 637                     Oscillospirales                     Butyricicoccaceae
## 638                       Eubacteriales                        Eubacteriaceae
## 639                       Eubacteriales                        Eubacteriaceae
## 640                      Lachnospirales                       Lachnospiraceae
## 641                      Lachnospirales                       Lachnospiraceae
## 642                      Lachnospirales                       Lachnospiraceae
## 643                      Lachnospirales                       Lachnospiraceae
## 644                      Lachnospirales                       Lachnospiraceae
## 645                      Lachnospirales                       Lachnospiraceae
## 646                      Lachnospirales                       Lachnospiraceae
## 647                      Lachnospirales                       Lachnospiraceae
## 648                      Lachnospirales                       Lachnospiraceae
## 649                      Lachnospirales                       Lachnospiraceae
## 650 Peptostreptococcales-Tissierellales                 Peptostreptococcaceae
## 651                          Clostridia                Hungateiclostridiaceae
## 652                        Monoglobales                         Monoglobaceae
## 653                          Clostridia                Hungateiclostridiaceae
## 654                     Oscillospirales                      Oscillospiraceae
## 655                     Oscillospirales                      Oscillospiraceae
## 656                     Oscillospirales                      Oscillospiraceae
## 657                     Oscillospirales                     Butyricicoccaceae
## 658                     Oscillospirales                      Oscillospiraceae
## 659                   Bacteriovoracales                    Bacteriovoracaceae
## 660                   Bacteriovoracales                    Bacteriovoracaceae
## 661                   Abditibacteriales                    Abditibacteriaceae
## 662                     Oscillospirales [Eubacterium]_coprostanoligenes_group
## 663                     Oscillospirales                       Ruminococcaceae
## 664                     Oscillospirales                       Ruminococcaceae
## 665                     Oscillospirales                       Ruminococcaceae
## 666                     Oscillospirales                    Ethanoligenenaceae
## 667                     Oscillospirales                       Ruminococcaceae
## 668                     Oscillospirales                       Oscillospirales
## 669                     Oscillospirales                       Ruminococcaceae
## 670                     Oscillospirales                       Ruminococcaceae
## 671                     Oscillospirales                       Ruminococcaceae
## 672                     Oscillospirales                     Butyricicoccaceae
## 673                      Lachnospirales                       Lachnospiraceae
## 674                     Oscillospirales                       Ruminococcaceae
## 675                     Oscillospirales                      Oscillospiraceae
## 676                     Oscillospirales                      Oscillospiraceae
## 677                     Oscillospirales                       Ruminococcaceae
## 678                     Oscillospirales                       Ruminococcaceae
## 679                     Oscillospirales                       Ruminococcaceae
## 680                     Oscillospirales    [Clostridium]_methylpentosum_group
## 681                       Clostridiales                        Clostridiaceae
## 682                  Clostridia_UCG-014                    Clostridia_UCG-014
## 683                      Lachnospirales                       Lachnospiraceae
## 684                Desulfitobacteriales                            uncultured
## 685                  Desulfotomaculales                    Desulfotomaculales
## 686                  Desulfotomaculales                    Desulfotomaculales
## 687                      Thermincolales                       Thermincolaceae
## 688                Desulfitobacteriales                     Heliobacteriaceae
## 689                Desulfitobacteriales                 Desulfitobacteriaceae
## 690                     Oscillospirales                            uncultured
## 691                       Peptococcales                        Peptococcaceae
## 692                       Peptococcales                        Peptococcaceae
## 693      Veillonellales-Selenomonadales                         Sporomusaceae
## 694                            BRH-c20a                              BRH-c20a
## 695                     Oscillospirales                               UCG-011
## 696                       Bacteroidales                        Marinifilaceae
## 697                    Tepidisphaerales                  CPla-3_termite_group
## 698                        Cytophagales                     Cyclobacteriaceae
## 699                        Cytophagales                       Amoebophilaceae
## 700                        Cytophagales                       Amoebophilaceae
## 701                       Bacteroidales                        Prevotellaceae
## 702                         Rhizobiales                      Beijerinckiaceae
## 703                       Clostridiales                            uncultured
## 704                       Clostridiales                      Caloramatoraceae
## 705                       Clostridiales                        Clostridiaceae
## 706                       Clostridiales                        Clostridiaceae
## 707                       Clostridiales                        Clostridiaceae
## 708                       Clostridiales                        Clostridiaceae
## 709                       MAT-CR-H4-C10                         MAT-CR-H4-C10
## 710                       Clostridiales                        Clostridiaceae
## 711                       Clostridiales                        Clostridiaceae
## 712                       Clostridiales                        Clostridiaceae
## 713                       Clostridiales                        Clostridiaceae
## 714                       Clostridiales                        Clostridiaceae
## 715                          Clostridia                Hungateiclostridiaceae
## 716                       Clostridiales                        Clostridiaceae
## 717                       Clostridiales                        Clostridiaceae
## 718                          Clostridia                Hungateiclostridiaceae
## 719                    Margulisbacteria                      Margulisbacteria
## 720                              DTU014                                DTU014
## 721                          uncultured                            uncultured
## 722                  Christensenellales                   Christensenellaceae
## 723                  Christensenellales                   Christensenellaceae
## 724                          Clostridia                    Gracilibacteraceae
## 725               Thermoactinomycetales                Thermoactinomycetaceae
## 726               Thermoactinomycetales                Thermoactinomycetaceae
## 727                    Natranaerobiales                     Natranaerobiaceae
## 728                     Lactobacillales                       Enterococcaceae
## 729 Peptostreptococcales-Tissierellales                 Peptostreptococcaceae
## 730                          Bacillales                           Bacillaceae
## 731                          Bacillales                        Planococcaceae
## 732 Peptostreptococcales-Tissierellales                 Peptostreptococcaceae
## 733 Peptostreptococcales-Tissierellales                 Peptostreptococcaceae
## 734 Peptostreptococcales-Tissierellales                 Peptostreptococcaceae
## 735 Peptostreptococcales-Tissierellales                 Peptostreptococcaceae
## 736 Peptostreptococcales-Tissierellales                      Anaerovoracaceae
## 737 Peptostreptococcales-Tissierellales                      Anaerovoracaceae
## 738 Peptostreptococcales-Tissierellales                      Anaerovoracaceae
## 739 Peptostreptococcales-Tissierellales                      Anaerovoracaceae
## 740 Peptostreptococcales-Tissierellales                      Anaerovoracaceae
## 741                        Limnochordia                          Limnochordia
## 742                          Clostridia                    Gracilibacteraceae
## 743                          Clostridia                Hungateiclostridiaceae
## 744                  Desulfotomaculales                    Desulfotomaculales
## 745                          Clostridia                Hungateiclostridiaceae
## 746                        Polyangiales                       Sandaracinaceae
## 747                           P9X2b3D02                             P9X2b3D02
## 748                   Campylobacterales                       Arcobacteraceae
## 749                       Rickettsiales                        Rickettsiaceae
## 750                       Reyranellales                        Reyranellaceae
## 751                  Diplorickettsiales                   Diplorickettsiaceae
## 752                  Diplorickettsiales                   Diplorickettsiaceae
## 753          Clostridia_vadinBB60_group            Clostridia_vadinBB60_group
## 754                Desulfitobacteriales                  Desulfitobacteriales
## 755                      Lachnospirales                       Lachnospiraceae
## 756                      Lachnospirales                       Lachnospiraceae
## 757                       Eubacteriales                        Eubacteriaceae
## 758                  Erysipelotrichales                   Erysipelotrichaceae
## 759                  Erysipelotrichales                            uncultured
## 760 Peptostreptococcales-Tissierellales                  Sedimentibacteraceae
## 761                      Lachnospirales                     Defluviitaleaceae
## 762                      Lachnospirales                       Lachnospiraceae
## 763                      Lachnospirales                       Lachnospiraceae
## 764                    Izemoplasmatales                      Izemoplasmatales
## 765                     Mycoplasmatales                      Mycoplasmataceae
## 766                    Coriobacteriales                       Eggerthellaceae
## 767                    Coriobacteriales                       Eggerthellaceae
## 768                    Coriobacteriales       Coriobacteriales_Incertae_Sedis
## 769                    Coriobacteriales                       Eggerthellaceae
## 770                    Coriobacteriales       Coriobacteriales_Incertae_Sedis
## 771                      Lachnospirales                       Lachnospiraceae
## 772                   Acidaminococcales                    Acidaminococcaceae
## 773      Veillonellales-Selenomonadales                         Sporomusaceae
## 774      Veillonellales-Selenomonadales                         Sporomusaceae
## 775      Veillonellales-Selenomonadales                         Sporomusaceae
## 776      Veillonellales-Selenomonadales                         Sporomusaceae
## 777                   Acidaminococcales                    Acidaminococcaceae
## 778      Veillonellales-Selenomonadales                         Sporomusaceae
## 779                Desulfitobacteriales                  Desulfitobacteriales
## 780                          Bacillales                        Planococcaceae
## 781                     Lactobacillales                     Carnobacteriaceae
## 782                       Victivallales                        Victivallaceae
## 783                              SJA-15                                SJA-15
## 784                     Lactobacillales                     Carnobacteriaceae
## 785                  Aneurinibacillales                   Aneurinibacillaceae
## 786                     Paenibacillales                      Paenibacillaceae
## 787                     Paenibacillales                      Paenibacillaceae
## 788                     Brevibacillales                      Brevibacillaceae
## 789                          Bacillales                       Marinococcaceae
## 790                          Bacillales                           Bacillaceae
## 791                          Bacillales                           Bacillaceae
## 792                          Bacillales                 Sporolactobacillaceae
## 793                          Bacillales                           Bacillaceae
## 794                    Enterobacterales                    Enterobacteriaceae
## 795                                RF39                                  RF39
## 796                  Vampirovibrionales                   Vampirovibrionaceae
## 797                  Vampirovibrionales                   Vampirovibrionaceae
## 798                   Obscuribacterales                    Obscuribacteraceae
## 799                     Phormidesmiales                       Nodosilineaceae
## 800                     Synechococcales                          Cyanobiaceae
## 801                    Cyanobacteriales                  Chroococcidiopsaceae
## 802                    Cyanobacteriales                            uncultured
## 803                    Cyanobacteriales                  Chroococcidiopsaceae
## 804                    Cyanobacteriales                  Chroococcidiopsaceae
## 805     Oxyphotobacteria_Incertae_Sedis                        Unknown_Family
## 806                    Pseudanabaenales                     Pseudanabaenaceae
## 807     Oxyphotobacteria_Incertae_Sedis                        Unknown_Family
## 808                    Cyanobacteriales                     Paraspirulinaceae
## 809     Oxyphotobacteria_Incertae_Sedis                        Unknown_Family
## 810                     Leptolyngbyales                      Leptolyngbyaceae
## 811                    Cyanobacteriales                           Nostocaceae
## 812                    Cyanobacteriales                    Coleofasciculaceae
## 813                    Cyanobacteriales                    Coleofasciculaceae
## 814                    Cyanobacteriales                         Phormidiaceae
## 815                    Cyanobacteriales                           Nostocaceae
## 816                     Lactobacillales                      Leuconostocaceae
## 817                  Erysipelotrichales             Erysipelatoclostridiaceae
## 818                  Erysipelotrichales             Erysipelatoclostridiaceae
## 819                  Erysipelotrichales                   Erysipelotrichaceae
## 820                  Erysipelotrichales             Erysipelatoclostridiaceae
## 821                  Erysipelotrichales             Erysipelatoclostridiaceae
## 822                  Erysipelotrichales             Erysipelatoclostridiaceae
## 823                   Acholeplasmatales                    Acholeplasmataceae
## 824                   Acholeplasmatales                    Acholeplasmataceae
## 825                   Acholeplasmatales                    Acholeplasmataceae
## 826                   Entomoplasmatales                    Entomoplasmataceae
## 827                   Entomoplasmatales                    Entomoplasmataceae
## 828                   Entomoplasmatales                     Spiroplasmataceae
## 829                            RsaHf231                              RsaHf231
## 830                  Erysipelotrichales                   Erysipelotrichaceae
## 831                  Erysipelotrichales                   Erysipelotrichaceae
## 832                  Erysipelotrichales                   Erysipelotrichaceae
## 833                   Entomoplasmatales      Entomoplasmatales_Incertae_Sedis
## 834                  Erysipelotrichales                   Erysipelotrichaceae
## 835                  Erysipelotrichales                   Erysipelotrichaceae
## 836                  Erysipelotrichales                   Erysipelotrichaceae
## 837                  Erysipelotrichales                   Erysipelotrichaceae
## 838                   Entomoplasmatales                     Entomoplasmatales
## 839                   Entomoplasmatales                              type_III
## 840                       Clostridiales                        Oxobacteraceae
## 841 Peptostreptococcales-Tissierellales                      Anaerovoracaceae
## 842                   Deferribacterales                    Deferribacteraceae
## 843                       Synergistales                        Synergistaceae
## 844                       Synergistales                        Synergistaceae
## 845                        Myxococcales                 Anaeromyxobacteraceae
## 846                     Paenibacillales                      Paenibacillaceae
## 847                     Mycoplasmatales                      Mycoplasmataceae
## 848                     Xanthomonadales                      Xanthomonadaceae
## 849                     Xanthomonadales                    Rhodanobacteraceae
## 850                     Xanthomonadales                      Xanthomonadaceae
## 851                     Xanthomonadales                    Rhodanobacteraceae
## 852                     Xanthomonadales                      Xanthomonadaceae
## 853                  Desulfovibrionales                   Desulfovibrionaceae
## 854                          uncultured                            uncultured
## 855                     Burkholderiales                      Aquaspirillaceae
## 856                     Burkholderiales                        Alcaligenaceae
## 857                     Burkholderiales                    Chitinibacteraceae
## 858                     Burkholderiales                     Nitrosomonadaceae
## 859                     Burkholderiales                     Nitrosomonadaceae
## 860                     Burkholderiales                        Rhodocyclaceae
## 861                     Burkholderiales                         Neisseriaceae
## 862                               R7C24                                 R7C24
## 863                     Pseudomonadales                         Moraxellaceae
## 864                     Burkholderiales                                   T34
## 865                    Enterobacterales                        Morganellaceae
## 866                    Enterobacterales                        Morganellaceae
## 867                    Enterobacterales                        Morganellaceae
## 868                    Enterobacterales                        Morganellaceae
## 869                      Pasteurellales                       Pasteurellaceae
## 870                     Cellvibrionales                      Cellvibrionaceae
## 871                   Oceanospirillales                   Pseudohongiellaceae
## 872                     Burkholderiales                      Methylophilaceae
## 873                     Burkholderiales                      Methylophilaceae
## 874                     Burkholderiales                        Rhodocyclaceae
## 875                     Burkholderiales                     Nitrosomonadaceae
## 876                     Burkholderiales                     Nitrosomonadaceae
## 877                     Burkholderiales                     Nitrosomonadaceae
## 878                    Enterobacterales                           Erwiniaceae
## 879                    Catenulisporales                       Actinospicaceae
## 880                   Corynebacteriales                      Mycobacteriaceae
## 881                       Eubacteriales                       Anaerofustaceae
## 882                       Eubacteriales                        Eubacteriaceae
## 883                        Chlamydiales                                  cvE6
## 884                         Subgroup_25                           Subgroup_25
## 885                  Christensenellales                   Christensenellaceae
## 886                       Bacteroidales                        Bacteroidaceae
## 887                       Bacteroidales                        Prevotellaceae
## 888                      Lachnospirales                       Lachnospiraceae
## 889                      Lachnospirales                       Lachnospiraceae
## 890                      Lachnospirales                       Lachnospiraceae
## 891                Desulfitobacteriales                   Syntrophobotulaceae
## 892                  Vicinamibacterales                   Vicinamibacteraceae
## 893                  Vicinamibacterales                   Vicinamibacteraceae
## 894                     Caulobacterales                       Hyphomonadaceae
## 895                     Pseudomonadales                         Moraxellaceae
## 896                     Fusobacteriales                      Fusobacteriaceae
## 897                     Burkholderiales                        Comamonadaceae
## 898                         Rhizobiales                     Xanthobacteraceae
## 899                         Rhizobiales                          Rhizobiaceae
## 900                              4-29-1                                4-29-1
## 901                     Burkholderiales                      Burkholderiaceae
## 902                    Gemmatimonadales                     Gemmatimonadaceae
## 903                          Frankiales                   Geodermatophilaceae
## 904                         Rhizobiales                      Beijerinckiaceae
## 905                   Thermomicrobiales                              AKYG1722
## 906                           Thermales                            Thermaceae
## 907 Peptostreptococcales-Tissierellales                 Peptostreptococcaceae
## 908                     Pseudomonadales                         Moraxellaceae
## 909                  Erysipelotrichales                   Erysipelotrichaceae
## 910                       Victivallales                             vadinBE97
## 911                           P.palmC41                             P.palmC41
## 912                     Oscillospirales                       Ruminococcaceae
## 913                       Bacteroidales                         Rikenellaceae
## 914                       Bacteroidales                         Rikenellaceae
## 915                       Bacteroidales                         Rikenellaceae
## 916                       Bacteroidales                         Rikenellaceae
## 917                       Bacteroidales                         Rikenellaceae
## 918                       Bacteroidales                    Prolixibacteraceae
## 919                     Oscillospirales                       Ruminococcaceae
## 920                       Bacteroidales                     Dysgonomonadaceae
## 921                       Bacteroidales                        Tannerellaceae
## 922                       Bacteroidales                        Tannerellaceae
## 923                       Bacteroidales                     Dysgonomonadaceae
## 924                       Bacteroidales                        Tannerellaceae
## 925                       Bacteroidales                        Tannerellaceae
## 926                       Bacteroidales                  Rs-E47_termite_group
## 927                       Bacteroidales                 Bacteroidales_UCG-001
## 928                       Bacteroidales                        Prevotellaceae
## 929                       Bacteroidales                     Dysgonomonadaceae
## 930                        Chromatiales                         Chromatiaceae
## 931                     Oscillospirales                       Ruminococcaceae
## 932                          Bacillales                           Bacillaceae
## 933                           0319-6G20                             0319-6G20
## 934                     Rhodobacterales                      Rhodobacteraceae
## 935                       Clostridiales                        Clostridiaceae
## 936                         Rhizobiales                      Beijerinckiaceae
## 937                       Bacteroidales              Bacteroidales_RF16_group
## 938                      Lachnospirales                       Lachnospiraceae
## 939                       Bacteroidales                        Marinifilaceae
## 940                       Bacteroidales                        Marinifilaceae
## 941                   Competibacterales                    Competibacteraceae
## 942                      Lachnospirales                       Lachnospiraceae
## 943      Veillonellales-Selenomonadales                      Selenomonadaceae
## 944                  Erysipelotrichales                   Erysipelotrichaceae
## 945                   Acidaminococcales                    Acidaminococcaceae
## 946      Veillonellales-Selenomonadales                            uncultured
## 947                   Acidaminococcales                    Acidaminococcaceae
## 948                  Erysipelotrichales             Erysipelatoclostridiaceae
## 949                      Lachnospirales                       Lachnospiraceae
## 950                      Lachnospirales                       Lachnospiraceae
## 951                      Lachnospirales                       Lachnospiraceae
## 952                       Bacteroidales                     Marinilabiliaceae
## 953                              KD4-96                                KD4-96
## 954                    Elusimicrobiales                     Elusimicrobiaceae
## 955                 Gastranaerophilales                   Gastranaerophilales
## 956               Thermosynechococcales                Thermosynechococcaceae
## 957                      Lachnospirales                       Lachnospiraceae
## 958                     Oscillospirales                       Ruminococcaceae
## 959                  Chthoniobacterales                   Chthoniobacteraceae
## 960                  Verrucomicrobiales                       Akkermansiaceae
##                                                  Genus Species
## 1                                           uncultured    <NA>
## 2                                      Thiophaeococcus    <NA>
## 3                                       Syntrophomonas    <NA>
## 4                                           uncultured    <NA>
## 5                                           uncultured    <NA>
## 6                                         Gemmatimonas    <NA>
## 7                                        Herpetosiphon    <NA>
## 8                                         ADurb.Bin180    <NA>
## 9                                           uncultured    <NA>
## 10                                         Kouleothrix    <NA>
## 11                                         Roseiflexus    <NA>
## 12                                         RBG-13-54-9    <NA>
## 13                                          uncultured    <NA>
## 14                                          Flexilinea    <NA>
## 15                                             SBR1031    <NA>
## 16                                                 A4b    <NA>
## 17                                        JG30-KF-CM45    <NA>
## 18                                                S085    <NA>
## 19                                        JG30-KF-CM66    <NA>
## 20                                          uncultured    <NA>
## 21                                         Litorilinea    <NA>
## 22                                                1-20    <NA>
## 23                                         JG30-KF-AS9    <NA>
## 24                                          Chloronema    <NA>
## 25                              Candidatus_Chloroploca    <NA>
## 26                                      Oscillochloris    <NA>
## 27                                            FFCH7168    <NA>
## 28                                              SHA-26    <NA>
## 29                                        Chthonomonas    <NA>
## 30                                           Elev-1554    <NA>
## 31                                         Gitt-GS-136    <NA>
## 32                                             AKIW781    <NA>
## 33                                               OLB14    <NA>
## 34                                        HSB_OF53-F07    <NA>
## 35                                              1959-1    <NA>
## 36                                       Ktedonobacter    <NA>
## 37                                    Thermosporothrix    <NA>
## 38                                          uncultured    <NA>
## 39                                             FCPS473    <NA>
## 40                                         JG30a-KF-32    <NA>
## 41                                          BacC-u-018    <NA>
## 42                                           G12-WMSP1    <NA>
## 43                                     Rokubacteriales    <NA>
## 44                                           B12-WMSP1    <NA>
## 45                                               GAL15    <NA>
## 46                                                TK10    <NA>
## 47                                               WPS-2    <NA>
## 48                                           Brevinema    <NA>
## 49                         Absconditabacteriales_(SR1)    <NA>
## 50                                          uncultured    <NA>
## 51                                        Elev-16S-573    <NA>
## 52                                       Blastocatella    <NA>
## 53                                                RB41    <NA>
## 54                                   Stenotrophobacter    <NA>
## 55                                         Lineage_IIc    <NA>
## 56                                         Lineage_IIa    <NA>
## 57                                         Lineage_IIb    <NA>
## 58                                          Luteimonas    <NA>
## 59                                       Paludibaculum    <NA>
## 60                               Candidatus_Solibacter    <NA>
## 61                                              GOUTB8    <NA>
## 62                                          Nitrospira    <NA>
## 63                                              KD3-10    <NA>
## 64                                           Bryocella    <NA>
## 65                                           Acidipila    <NA>
## 66                                          uncultured    <NA>
## 67                                     Occallatibacter    <NA>
## 68                                          uncultured    <NA>
## 69                                           Sumerlaea    <NA>
## 70                                   Longimicrobiaceae    <NA>
## 71                                      Longimicrobium    <NA>
## 72                                         Subgroup_17    <NA>
## 73                                              MVP-15    <NA>
## 74                                         bacteriap25    <NA>
## 75                        SAR324_clade(Marine_group_B)    <NA>
## 76                                       Sphaerochaeta    <NA>
## 77                                           Treponema    <NA>
## 78                                          uncultured    <NA>
## 79                                          uncultured    <NA>
## 80                                         Turneriella    <NA>
## 81                                          uncultured    <NA>
## 82                                          uncultured    <NA>
## 83                                        Conexibacter    <NA>
## 84                                          uncultured    <NA>
## 85                                       Ilumatobacter    <NA>
## 86                               CL500-29_marine_group    <NA>
## 87                                  Ilumatobacteraceae    <NA>
## 88                                          uncultured    <NA>
## 89                                               Iamia    <NA>
## 90                                          uncultured    <NA>
## 91                                           IMCC26256    <NA>
## 92                                          uncultured    <NA>
## 93                                      Microtrichales    <NA>
## 94                                           IMCC26207    <NA>
## 95                                     Propioniciclava    <NA>
## 96                                  Aestuariimicrobium    <NA>
## 97                                    Marinilutecoccus    <NA>
## 98                                             Euzebya    <NA>
## 99                                          uncultured    <NA>
## 100                                   Scytonema_UCFS19    <NA>
## 101                                    Actinoallomurus    <NA>
## 102                                            Gaiella    <NA>
## 103                                       Patulibacter    <NA>
## 104                                          JCM_18997    <NA>
## 105                                    Solirubrobacter    <NA>
## 106                                     Terrimicrobium    <NA>
## 107                                         uncultured    <NA>
## 108                                   Parviterribacter    <NA>
## 109                                         uncultured    <NA>
## 110                                         Bryobacter    <NA>
## 111                                              67-14    <NA>
## 112                                          vadinHA49    <NA>
## 113                                         uncultured    <NA>
## 114                                              BSV26    <NA>
## 115                                       Zixibacteria    <NA>
## 116                                               B122    <NA>
## 117                                             BBMC-4    <NA>
## 118                                         SR-FBR-L83    <NA>
## 119                                          PHOS-HE36    <NA>
## 120                                    Kapabacteriales    <NA>
## 121                                             SJA-28    <NA>
## 122                                         uncultured    <NA>
## 123                                         Rubrivirga    <NA>
## 124                                   Chthonomonadales    <NA>
## 125                                    Armatimonadales    <NA>
## 126                                        Armatimonas    <NA>
## 127                                         uncultured    <NA>
## 128                                  Xanthobacteraceae    <NA>
## 129                                         uncultured    <NA>
## 130                                         KF-JG30-B3    <NA>
## 131                                              A0839    <NA>
## 132                                         uncultured    <NA>
## 133                                         uncultured    <NA>
## 134                                         uncultured    <NA>
## 135                                       Geminicoccus    <NA>
## 136                           Candidatus_Alysiosphaera    <NA>
## 137                                         uncultured    <NA>
## 138                                             Dongia    <NA>
## 139                                     Defluviicoccus    <NA>
## 140                                        Skermanella    <NA>
## 141                                        Rhodovarius    <NA>
## 142                                         Roseomonas    <NA>
## 143                                        Rubritepida    <NA>
## 144                                         uncultured    <NA>
## 145                                       Acidiphilium    <NA>
## 146                                         uncultured    <NA>
## 147                                          Acidisoma    <NA>
## 148                                        Rhodovastum    <NA>
## 149                                        Acidicaldus    <NA>
## 150                                          Rhodopila    <NA>
## 151                                         uncultured    <NA>
## 152                                         uncultured    <NA>
## 153                                         uncultured    <NA>
## 154                          Candidatus_Anadelfobacter    <NA>
## 155                             Candidatus_Jidaibacter    <NA>
## 156                                             SM2D12    <NA>
## 157                                         uncultured    <NA>
## 158                                   Polymorphobacter    <NA>
## 159                                 Sandarakinorhabdus    <NA>
## 160                                       Sphingomonas    <NA>
## 161                                        Sphingobium    <NA>
## 162                                   Telmatospirillum    <NA>
## 163                                             DSSF69    <NA>
## 164                                    Novosphingobium    <NA>
## 165                                 Altererythrobacter    <NA>
## 166                                        Rhizorhapis    <NA>
## 167                                        1174-901-12    <NA>
## 168                                             Labrys    <NA>
## 169                     Methylobacterium-Methylorubrum    <NA>
## 170                                      Phreatobacter    <NA>
## 171                                             Afipia    <NA>
## 172                                         uncultured    <NA>
## 173                                            FukuN57    <NA>
## 174                                       Pseudolabrys    <NA>
## 175                                 Methyloligellaceae    <NA>
## 176                                            Bauldia    <NA>
## 177                                   Rubellimicrobium    <NA>
## 178                                        Camelimonas    <NA>
## 179                                         Roseiarcus    <NA>
## 180                                      Brevundimonas    <NA>
## 181                                              PMMR1    <NA>
## 182                                   Phenylobacterium    <NA>
## 183                                         uncultured    <NA>
## 184                                       Ancylobacter    <NA>
## 185                                       Xanthobacter    <NA>
## 186                                  Pseudorhodoplanes    <NA>
## 187                                     Methylovirgula    <NA>
## 188                                            Devosia    <NA>
## 189                                         uncultured    <NA>
## 190                                           Nordella    <NA>
## 191                                     Rhodomicrobium    <NA>
## 192                                     Hyphomicrobium    <NA>
## 193                                         uncultured    <NA>
## 194                                            Kaistia    <NA>
## 195                                      Pedomicrobium    <NA>
## 196                                         uncultured    <NA>
## 197                                  Psychroglaciecola    <NA>
## 198                                         Microvirga    <NA>
## 199                                           Hirschia    <NA>
## 200                                              D05-2    <NA>
## 201                                         uncultured    <NA>
## 202                                         uncultured    <NA>
## 203                                Candidatus_Captivus    <NA>
## 204                                         uncultured    <NA>
## 205                                                AB1    <NA>
## 206                                      Holosporaceae    <NA>
## 207                                  Sericytochromatia    <NA>
## 208                                      Neomegalonema    <NA>
## 209                                    Ignatzschineria    <NA>
## 210                                          Koukoulia    <NA>
## 211                                          Aquicella    <NA>
## 212                           Candidatus_Competibacter    <NA>
## 213                                         uncultured    <NA>
## 214                                              HOC36    <NA>
## 215                                      Sulfurifustis    <NA>
## 216                                         uncultured    <NA>
## 217                                         uncultured    <NA>
## 218                                              CCD24    <NA>
## 219                                             PLTA13    <NA>
## 220                                         uncultured    <NA>
## 221                                            Nevskia    <NA>
## 222                                       JG36-TzT-191    <NA>
## 223                                           Coxiella    <NA>
## 224                                         JG36-GS-52    <NA>
## 225                                     Steroidobacter    <NA>
## 226                                         OM27_clade    <NA>
## 227                                         uncultured    <NA>
## 228                                          Halomonas    <NA>
## 229                                               HdN1    <NA>
## 230                                            wb1-P19    <NA>
## 231                                        KF-JG30-C25    <NA>
## 232                                          Thiothrix    <NA>
## 233                                        Subgroup_10    <NA>
## 234                                  Methyloparacoccus    <NA>
## 235                                         CHAB-XI-27    <NA>
## 236                                Gammaproteobacteria    <NA>
## 237                               Candidatus_Berkiella    <NA>
## 238                            Candidatus_Ovatusbacter    <NA>
## 239                                     Unknown_Family    <NA>
## 240                                       F9P41300-M23    <NA>
## 241                                         uncultured    <NA>
## 242                                         Crenothrix    <NA>
## 243                                             MBNT15    <NA>
## 244                                            RCP2-54    <NA>
## 245                                           Truepera    <NA>
## 246                                        Deinococcus    <NA>
## 247                                         uncultured    <NA>
## 248                                   Syntrophorhabdus    <NA>
## 249                                       Bdellovibrio    <NA>
## 250                                         Syntrophus    <NA>
## 251                                         uncultured    <NA>
## 252                             Sva0081_sediment_group    <NA>
## 253                                          Geobacter    <NA>
## 254                                         uncultured    <NA>
## 255                                       Desulfobacca    <NA>
## 256                                      Silvanigrella    <NA>
## 257                                               WX65    <NA>
## 258                                             BD7-11    <NA>
## 259                                     Nannocystaceae    <NA>
## 260                                        Nannocystis    <NA>
## 261                                     Bradymonadales    <NA>
## 262                                         uncultured    <NA>
## 263                                        Oligoflexus    <NA>
## 264                                               PB19    <NA>
## 265                                            Blfdi19    <NA>
## 266                                              NKB15    <NA>
## 267                            Craurococcus-Caldovatus    <NA>
## 268                                        Cystobacter    <NA>
## 269                                      Vulgatibacter    <NA>
## 270                                         uncultured    <NA>
## 271                                      Phaselicystis    <NA>
## 272                                           MSB-4B10    <NA>
## 273                                         Haliangium    <NA>
## 274                                          VHS-B4-70    <NA>
## 275                                            mle1-27    <NA>
## 276                                      Aetherobacter    <NA>
## 277                                         Polyangium    <NA>
## 278                                   Pajaroellobacter    <NA>
## 279                                          Sorangium    <NA>
## 280                                         uncultured    <NA>
## 281                                         uncultured    <NA>
## 282                                            BIrii41    <NA>
## 283                                              NB1-j    <NA>
## 284                                          FFCH16767    <NA>
## 285                                         uncultured    <NA>
## 286                              Candidatus_Koribacter    <NA>
## 287                                         Subgroup_2    <NA>
## 288                                        Subgroup_11    <NA>
## 289                                   Latescibacterota    <NA>
## 290                                              11-24    <NA>
## 291                                         uncultured    <NA>
## 292                                   Tellurimicrobium    <NA>
## 293                                    Gracilibacteria    <NA>
## 294                                    JGI_0000069-P22    <NA>
## 295                                       Methanolinea    <NA>
## 296                                 Methanobrevibacter    <NA>
## 297                                   Methanobacterium    <NA>
## 298                       Candidatus_Magasanikbacteria    <NA>
## 299                        Candidatus_Yanofskybacteria    <NA>
## 300                                      Parcubacteria    <NA>
## 301                                  Saccharimonadales    <NA>
## 302                                       Fimbriimonas    <NA>
## 303                                  Fimbriimonadaceae    <NA>
## 304                                               LWQ8    <NA>
## 305                                              WWH38    <NA>
## 306                                                TM7    <NA>
## 307                                               TM7a    <NA>
## 308                                        Pseudomonas    <NA>
## 309                                       Helicobacter    <NA>
## 310                             Candidatus_Omnitrophus    <NA>
## 311                                         uncultured    <NA>
## 312                                      Campylobacter    <NA>
## 313                                         uncultured    <NA>
## 314                                       Sulfurimonas    <NA>
## 315                                    Pseudarcobacter    <NA>
## 316                                 Nitrososphaeraceae    <NA>
## 317                                   Nitrosotaleaceae    <NA>
## 318                                       Methanosaeta    <NA>
## 319                                         Halomarina    <NA>
## 320                                 Methanocorpusculum    <NA>
## 321                           Candidatus_Methanoplasma    <NA>
## 322                                    Woesearchaeales    <NA>
## 323                                           UBA12409    <NA>
## 324                                        Babeliaceae    <NA>
## 325                                     Vermiphilaceae    <NA>
## 326                                         Babeliales    <NA>
## 327                                      Holophagaceae    <NA>
## 328                                      Lacunisphaera    <NA>
## 329                                         uncultured    <NA>
## 330                                         uncultured    <NA>
## 331                                         uncultured    <NA>
## 332                                       Cerasicoccus    <NA>
## 333                                       Alterococcus    <NA>
## 334                                     ADurb.Bin063-1    <NA>
## 335                                    Pedosphaeraceae    <NA>
## 336                       Candidatus_Xiphinematobacter    <NA>
## 337                             Candidatus_Udaeobacter    <NA>
## 338                                         uncultured    <NA>
## 339                                         uncultured    <NA>
## 340                                      Luteolibacter    <NA>
## 341                                             DEV114    <NA>
## 342                                        Pedosphaera    <NA>
## 343                                             DEV008    <NA>
## 344                                             SH3-11    <NA>
## 345                                         Oikopleura    <NA>
## 346                                S-BQ2-57_soil_group    <NA>
## 347                                            01D2Z36    <NA>
## 348                                               LD29    <NA>
## 349                                               UA11    <NA>
## 350                                     Roseimicrobium    <NA>
## 351                                    Prosthecobacter    <NA>
## 352                                   Verrucomicrobium    <NA>
## 353                                     Leptospirillum    <NA>
## 354                                        Subgroup_22    <NA>
## 355                                Vicinamibacteraceae    <NA>
## 356                                        Subgroup_18    <NA>
## 357                                           AT-s3-28    <NA>
## 358                                         Subgroup_7    <NA>
## 359                                  Entotheonellaceae    <NA>
## 360                                            P3OB-42    <NA>
## 361                                 Hydrogenedensaceae    <NA>
## 362                                       Pla4_lineage    <NA>
## 363                                Thermoanaerobaculum    <NA>
## 364                                            LD1-PB3    <NA>
## 365                                           WCHB1-41    <NA>
## 366                                             DS-100    <NA>
## 367                                       Tumebacillus    <NA>
## 368                           Candidatus_Metachlamydia    <NA>
## 369                                         uncultured    <NA>
## 370                                      Chlamydiaceae    <NA>
## 371                                       Neochlamydia    <NA>
## 372                          Candidatus_Protochlamydia    <NA>
## 373                       Candidatus_Amphibiichlamydia    <NA>
## 374                                          Ga0074140    <NA>
## 375                         Candidatus_Rhabdochlamydia    <NA>
## 376                           Candidatus_Mesochlamydia    <NA>
## 377                                           Estrella    <NA>
## 378                                         uncultured    <NA>
## 379                                     Guggenheimella    <NA>
## 380                                       Gottschalkia    <NA>
## 381                                         Fusibacter    <NA>
## 382                                      Peptoniphilus    <NA>
## 383                                              W5053    <NA>
## 384                                        Jonquetella    <NA>
## 385                                     Pyramidobacter    <NA>
## 386                      Christensenellaceae_R-7_group    <NA>
## 387                                              OPB41    <NA>
## 388                                         uncultured    <NA>
## 389                          Coriobacteriaceae_UCG-002    <NA>
## 390                                          Atopobium    <NA>
## 391                                         Lysobacter    <NA>
## 392                                           Ahniella    <NA>
## 393                                   Pseudofulvimonas    <NA>
## 394                                         Legionella    <NA>
## 395                                            C1-B045    <NA>
## 396                                        KI89A_clade    <NA>
## 397                                         uncultured    <NA>
## 398                                            FCPU426    <NA>
## 399                                              WD260    <NA>
## 400                               Thermoflavimicrobium    <NA>
## 401                                          Pasteuria    <NA>
## 402                                        Shimazuella    <NA>
## 403                                      Sh765B-TzT-35    <NA>
## 404                        Candidatus_Methylomirabilis    <NA>
## 405                                                AD3    <NA>
## 406                                    Dadabacteriales    <NA>
## 407                                         uncultured    <NA>
## 408                                       Paludibacter    <NA>
## 409                                               F082    <NA>
## 410                                           p-251-o5    <NA>
## 411                        Rikenellaceae_RC9_gut_group    <NA>
## 412                                            UCG-005    <NA>
## 413                                         uncultured    <NA>
## 414                                         uncultured    <NA>
## 415                                             DMER64    <NA>
## 416                                            AKYH767    <NA>
## 417                            Bacteroidetes_vadinHA17    <NA>
## 418                                       Hymenobacter    <NA>
## 419                                          Cytophaga    <NA>
## 420                                     Sporocytophaga    <NA>
## 421                                         uncultured    <NA>
## 422                                      Adhaeribacter    <NA>
## 423                                         uncultured    <NA>
## 424                                     Rhodocytophaga    <NA>
## 425                                    Ferruginibacter    <NA>
## 426                                       Ohtaekwangia    <NA>
## 427                                              OLB12    <NA>
## 428                                       Lacihabitans    <NA>
## 429                                         uncultured    <NA>
## 430                                          Emticicia    <NA>
## 431                    Blvii28_wastewater-sludge_group    <NA>
## 432                                Bacteroidetes_BD2-2    <NA>
## 433                                           ST-12K33    <NA>
## 434                     [Cytophaga]_xylanolytica_group    <NA>
## 435                                         uncultured    <NA>
## 436                                                 H1    <NA>
## 437                                              F0058    <NA>
## 438                                      Bacteroidales    <NA>
## 439                                     Muribaculaceae    <NA>
## 440                                        Coprobacter    <NA>
## 441                                         uncultured    <NA>
## 442                                              BSV13    <NA>
## 443                                          Solitalea    <NA>
## 444                                             Z4MB62    <NA>
## 445                                          vadinHA21    <NA>
## 446                                         env.OPS_17    <NA>
## 447                                   NS9_marine_group    <NA>
## 448                               NS11-12_marine_group    <NA>
## 449                                         uncultured    <NA>
## 450                                  Haliscomenobacter    <NA>
## 451                                   Aurantisolimonas    <NA>
## 452                                     Arachidicoccus    <NA>
## 453                                            Cnuella    <NA>
## 454                                   Chitinophagaceae    <NA>
## 455                                           Niabella    <NA>
## 456                                         Terrimonas    <NA>
## 457                                    Flavisolibacter    <NA>
## 458                                       Segetibacter    <NA>
## 459                                    Flavihumibacter    <NA>
## 460                                   Pseudoflavitalea    <NA>
## 461                                               Puia    <NA>
## 462                                         Flavitalea    <NA>
## 463                                      Edaphobaculum    <NA>
## 464                                         Taibaiella    <NA>
## 465                                   Pseudopedobacter    <NA>
## 466                                         Fluviicola    <NA>
## 467                               Rs-D38_termite_group    <NA>
## 468                                             CR-115    <NA>
## 469                                        Anaerocella    <NA>
## 470                                              PeH15    <NA>
## 471                             M2PB4-65_termite_group    <NA>
## 472                             COB_P4-1_termite_group    <NA>
## 473                                       Crocinitomix    <NA>
## 474                                     Lentimicrobium    <NA>
## 475                                  Lentimicrobiaceae    <NA>
## 476                                         uncultured    <NA>
## 477                                               SB-5    <NA>
## 478                                        Chishuiella    <NA>
## 479                                       Empedobacter    <NA>
## 480                                          Apibacter    <NA>
## 481                                   Chryseobacterium    <NA>
## 482                                     Flavobacterium    <NA>
## 483                                          Spirosoma    <NA>
## 484                                           Fibrella    <NA>
## 485                                        Dyadobacter    <NA>
## 486                                      Siphonobacter    <NA>
## 487                                     Pseudarcicella    <NA>
## 488                                          Arcicella    <NA>
## 489                                        Flexibacter    <NA>
## 490                                       Chryseolinea    <NA>
## 491                                         uncultured    <NA>
## 492                                   Sphingobacterium    <NA>
## 493                                     Parapedobacter    <NA>
## 494                                   Mucilaginibacter    <NA>
## 495                                           Myroides    <NA>
## 496                                     Capnocytophaga    <NA>
## 497                                            Gemmata    <NA>
## 498                                        Telmatocola    <NA>
## 499                                      Fimbriiglobus    <NA>
## 500                                       Planctopirus    <NA>
## 501                                      Paludisphaera    <NA>
## 502                                   Planctomicrobium    <NA>
## 503                                         uncultured    <NA>
## 504                                         uncultured    <NA>
## 505                                            SH-PL14    <NA>
## 506                                        Schlesneria    <NA>
## 507                                p-1088-a5_gut_group    <NA>
## 508                      Termite_planctomycete_cluster    <NA>
## 509                                          Pirellula    <NA>
## 510                                       Pir4_lineage    <NA>
## 511                                         uncultured    <NA>
## 512                                     Rhodopirellula    <NA>
## 513                       Candidatus_Anammoximicrobium    <NA>
## 514                                    Blastopirellula    <NA>
## 515                               CPla-4_termite_group    <NA>
## 516                                              OM190    <NA>
## 517                                               S-70    <NA>
## 518                                       Pla1_lineage    <NA>
## 519                                             SM1A02    <NA>
## 520                                            AKYG587    <NA>
## 521                                         uncultured    <NA>
## 522                                              SG8-4    <NA>
## 523                            AKAU3564_sediment_group    <NA>
## 524                                  WD2101_soil_group    <NA>
## 525                                  Tepidisphaeraceae    <NA>
## 526                                      Tepidisphaera    <NA>
## 527                                       Zavarzinella    <NA>
## 528                                         Tuwongella    <NA>
## 529                                         uncultured    <NA>
## 530                                         uncultured    <NA>
## 531                                        Gemmataceae    <NA>
## 532                                         Isosphaera    <NA>
## 533                                     Singulisphaera    <NA>
## 534                              Candidatus_Nostocoida    <NA>
## 535                                      Tundrisphaera    <NA>
## 536                                        Aquisphaera    <NA>
## 537                                         uncultured    <NA>
## 538                          Candidatus_Nomurabacteria    <NA>
## 539                                         uncultured    <NA>
## 540                                          Bilophila    <NA>
## 541                                         Subgroup_5    <NA>
## 542                                    Oscillospirales    <NA>
## 543                                      Pygmaiobacter    <NA>
## 544                                    Subdoligranulum    <NA>
## 545                                        Anaerofilum    <NA>
## 546                                      Streptococcus    <NA>
## 547                                        Floricoccus    <NA>
## 548                                        Rubrobacter    <NA>
## 549                                          MB-A2-108    <NA>
## 550                                 Micromonosporaceae    <NA>
## 551                                          FFCH16263    <NA>
## 552                                        Marmoricola    <NA>
## 553                                              PeM15    <NA>
## 554                                         uncultured    <NA>
## 555                                         uncultured    <NA>
## 556                                        Salinispora    <NA>
## 557                                      Phytohabitans    <NA>
## 558                                   Jatrophihabitans    <NA>
## 559                                          Tomitella    <NA>
## 560                                    Corynebacterium    <NA>
## 561                                         Lawsonella    <NA>
## 562                                     Smaragdicoccus    <NA>
## 563                                         uncultured    <NA>
## 564                                        Rhodococcus    <NA>
## 565                                  Corynebacteriales    <NA>
## 566                                   Cryptosporangium    <NA>
## 567                                        Antricoccus    <NA>
## 568                                      Quadrisphaera    <NA>
## 569                                        Kineosporia    <NA>
## 570                                        Actinomyces    <NA>
## 571                                             Rothia    <NA>
## 572                                     Arsenicicoccus    <NA>
## 573                                   Dermatophilaceae    <NA>
## 574                                       Mobilicoccus    <NA>
## 575                                     Aquipuribacter    <NA>
## 576                                             Salana    <NA>
## 577                                         uncultured    <NA>
## 578                                          Agromyces    <NA>
## 579                                        Kineococcus    <NA>
## 580                               Candidatus_Ancillula    <NA>
## 581                                      Angustibacter    <NA>
## 582                                         uncultured    <NA>
## 583                                      Catenulispora    <NA>
## 584                                         uncultured    <NA>
## 585                                         Crossiella    <NA>
## 586                                         E1B-B3-114    <NA>
## 587                                      Amycolatopsis    <NA>
## 588                                    Actinophytocola    <NA>
## 589                                       Actinoplanes    <NA>
## 590                                      Luedemannella    <NA>
## 591                                           Hamadaea    <NA>
## 592                                             Asanoa    <NA>
## 593                                         uncultured    <NA>
## 594                                       Streptomyces    <NA>
## 595                                         Longivirga    <NA>
## 596                                     Pseudonocardia    <NA>
## 597                                        Nakamurella    <NA>
## 598                                       Nocardioides    <NA>
## 599                                       Herbidospora    <NA>
## 600                                         uncultured    <NA>
## 601                                     Stackebrandtia    <NA>
## 602                                       Acidothermus    <NA>
## 603                                       Actinomadura    <NA>
## 604                                         Frankiales    <NA>
## 605                                     Actinocorallia    <NA>
## 606                                       Microlunatus    <NA>
## 607                                     Friedmanniella    <NA>
## 608                                      Aeromicrobium    <NA>
## 609                                        Marinomonas    <NA>
## 610                                        Limnobacter    <NA>
## 611                                          Lautropia    <NA>
## 612                                               MND1    <NA>
## 613                                         uncultured    <NA>
## 614                                       Sphaerotilus    <NA>
## 615                                        Lampropedia    <NA>
## 616                                              AAP99    <NA>
## 617                                         Collimonas    <NA>
## 618                                          Aquitalea    <NA>
## 619                                       Vitreoscilla    <NA>
## 620                                          Vogesella    <NA>
## 621                                        Crenobacter    <NA>
## 622                                       Thiobacillus    <NA>
## 623                                            SC-I-84    <NA>
## 624                                               A21b    <NA>
## 625                                         uncultured    <NA>
## 626                                      Stenoxybacter    <NA>
## 627                                            TRA3-20    <NA>
## 628                                             B1-7BS    <NA>
## 629                                      Lactobacillus    <NA>
## 630                                          Tolumonas    <NA>
## 631                                         uncultured    <NA>
## 632                            Lachnospiraceae_UCG-008    <NA>
## 633                         [Eubacterium]_hallii_group    <NA>
## 634                                      V9D2013_group    <NA>
## 635                                            UCG-010    <NA>
## 636                                      Papillibacter    <NA>
## 637                                            UCG-008    <NA>
## 638                                        Eubacterium    <NA>
## 639                                     Acetobacterium    <NA>
## 640                                   Cellulosilyticum    <NA>
## 641                                       Anaerostipes    <NA>
## 642                                        Lachnospira    <NA>
## 643                                    Lachnospiraceae    <NA>
## 644                    [Eubacterium]_ruminantium_group    <NA>
## 645                                     Frisingicoccus    <NA>
## 646                                     Catenibacillus    <NA>
## 647                       Lachnospiraceae_ND3007_group    <NA>
## 648                            Lachnospiraceae_UCG-006    <NA>
## 649                                      Anaerostignum    <NA>
## 650                                         uncultured    <NA>
## 651                                         uncultured    <NA>
## 652                                         Monoglobus    <NA>
## 653                                  Ruminiclostridium    <NA>
## 654                                            UCG-002    <NA>
## 655                                      Oscillibacter    <NA>
## 656                                       Oscillospira    <NA>
## 657                                     Butyricicoccus    <NA>
## 658                                      NK4A214_group    <NA>
## 659                                       Peredibacter    <NA>
## 660                                      Bacteriovorax    <NA>
## 661                                    Abditibacterium    <NA>
## 662              [Eubacterium]_coprostanoligenes_group    <NA>
## 663                                  Caproiciproducens    <NA>
## 664                                     Incertae_Sedis    <NA>
## 665                                         uncultured    <NA>
## 666                                     Incertae_Sedis    <NA>
## 667                                      Angelakisella    <NA>
## 668                           Hydrogenoanaerobacterium    <NA>
## 669                             Candidatus_Soleaferrea    <NA>
## 670                                   Faecalibacterium    <NA>
## 671                                    Ruminococcaceae    <NA>
## 672                                            UCG-009    <NA>
## 673                  [Eubacterium]_oxidoreducens_group    <NA>
## 674                                            UBA1819    <NA>
## 675                                     Intestinimonas    <NA>
## 676                                   Colidextribacter    <NA>
## 677                                      Anaerotruncus    <NA>
## 678                                         Paludicola    <NA>
## 679                                       Harryflintia    <NA>
## 680                 [Clostridium]_methylpentosum_group    <NA>
## 681                                     Clostridiaceae    <NA>
## 682                                 Clostridia_UCG-014    <NA>
## 683                                       Epulopiscium    <NA>
## 684                                         uncultured    <NA>
## 685                                    Desulfofarcimen    <NA>
## 686                                      Pelotomaculum    <NA>
## 687                                        Thermincola    <NA>
## 688                                         uncultured    <NA>
## 689                                  Desulfosporosinus    <NA>
## 690                                         uncultured    <NA>
## 691                                        Peptococcus    <NA>
## 692                                         uncultured    <NA>
## 693                                        Anaerosinus    <NA>
## 694                                           BRH-c20a    <NA>
## 695                                            UCG-011    <NA>
## 696                                      Butyricimonas    <NA>
## 697                               CPla-3_termite_group    <NA>
## 698                                      Tunicatimonas    <NA>
## 699                            Candidatus_Amoebophilus    <NA>
## 700                          Candidatus_Paenicardinium    <NA>
## 701                         Prevotellaceae_Ga6A1_group    <NA>
## 702                                       Rhodoblastus    <NA>
## 703                                         uncultured    <NA>
## 704                                         Fonticella    <NA>
## 705                                         uncultured    <NA>
## 706                        Clostridium_sensu_stricto_3    <NA>
## 707                        Clostridium_sensu_stricto_9    <NA>
## 708                       Clostridium_sensu_stricto_10    <NA>
## 709                                      MAT-CR-H4-C10    <NA>
## 710                                          Hathewaya    <NA>
## 711                       Clostridium_sensu_stricto_13    <NA>
## 712                             Candidatus_Arthromitus    <NA>
## 713                       Clostridium_sensu_stricto_11    <NA>
## 714                        Clostridium_sensu_stricto_2    <NA>
## 715                                        Acetivibrio    <NA>
## 716                                            Sarcina    <NA>
## 717                        Clostridium_sensu_stricto_1    <NA>
## 718                                     Fastidiosipila    <NA>
## 719                                   Margulisbacteria    <NA>
## 720                                             DTU014    <NA>
## 721                                         uncultured    <NA>
## 722                                    Christensenella    <NA>
## 723                                         uncultured    <NA>
## 724                                          Lutispora    <NA>
## 725                                  Thermoactinomyces    <NA>
## 726                                         uncultured    <NA>
## 727                                     Natranaerobius    <NA>
## 728                                       Enterococcus    <NA>
## 729                                   Peptoclostridium    <NA>
## 730                                      Terribacillus    <NA>
## 731                                       Sporosarcina    <NA>
## 732                                     Clostridioides    <NA>
## 733                                    Paraclostridium    <NA>
## 734                                       Tepidibacter    <NA>
## 735                                   Terrisporobacter    <NA>
## 736                         [Eubacterium]_brachy_group    <NA>
## 737                                         uncultured    <NA>
## 738                                Family_XIII_UCG-001    <NA>
## 739                                           Amnipila    <NA>
## 740                                        Anaerovorax    <NA>
## 741                                     Hydrogenispora    <NA>
## 742                                         uncultured    <NA>
## 743                                          HN-HF0106    <NA>
## 744                                     Desulfurispora    <NA>
## 745                             Hungateiclostridiaceae    <NA>
## 746                                       Sandaracinus    <NA>
## 747                                          P9X2b3D02    <NA>
## 748                                         Arcobacter    <NA>
## 749                                         uncultured    <NA>
## 750                                         Reyranella    <NA>
## 751                                    Diplorickettsia    <NA>
## 752                                      Rickettsiella    <NA>
## 753                         Clostridia_vadinBB60_group    <NA>
## 754                                                TC1    <NA>
## 755                                         uncultured    <NA>
## 756                                    Natranaerovirga    <NA>
## 757                                   Pseudoramibacter    <NA>
## 758                                             Dielma    <NA>
## 759                                         uncultured    <NA>
## 760                                    Sedimentibacter    <NA>
## 761                          Defluviitaleaceae_UCG-011    <NA>
## 762                                         Tyzzerella    <NA>
## 763                                  Anaerosporobacter    <NA>
## 764                                   Izemoplasmatales    <NA>
## 765                                         Mycoplasma    <NA>
## 766                                         uncultured    <NA>
## 767                                      Enterorhabdus    <NA>
## 768                                     Phoenicibacter    <NA>
## 769                                      Gordonibacter    <NA>
## 770                                         uncultured    <NA>
## 771                                          Hespellia    <NA>
## 772                                         uncultured    <NA>
## 773                                          Pelosinus    <NA>
## 774                                  Dendrosporobacter    <NA>
## 775                                          Sporomusa    <NA>
## 776                                         uncultured    <NA>
## 777                                    Acidaminococcus    <NA>
## 778                                        Anaerospora    <NA>
## 779                    Candidatus_Dichloromethanomonas    <NA>
## 780                                     Lysinibacillus    <NA>
## 781                                         uncultured    <NA>
## 782                                     Victivallaceae    <NA>
## 783                                             SJA-15    <NA>
## 784                                     Dolosigranulum    <NA>
## 785                                   Aneurinibacillus    <NA>
## 786                                       Ammoniphilus    <NA>
## 787                                           Cohnella    <NA>
## 788                                      Brevibacillus    <NA>
## 789                                       Geomicrobium    <NA>
## 790                                  Halolactibacillus    <NA>
## 791                              Pseudogracilibacillus    <NA>
## 792                                 Sporolactobacillus    <NA>
## 793                                     Oceanobacillus    <NA>
## 794                               Escherichia-Shigella    <NA>
## 795                                               RF39    <NA>
## 796                                      Vampirovibrio    <NA>
## 797                                Vampirovibrionaceae    <NA>
## 798                                 Obscuribacteraceae    <NA>
## 799                                          MBIC10086    <NA>
## 800                                 Cyanobium_PCC-6307    <NA>
## 801                                         Aliterella    <NA>
## 802                                         uncultured    <NA>
## 803                                         uncultured    <NA>
## 804                         Chroococcidiopsis_SAG_2023    <NA>
## 805                                            CENA359    <NA>
## 806                             Pseudanabaena_PCC-7429    <NA>
## 807                                         uncultured    <NA>
## 808                                         uncultured    <NA>
## 809                             Leptolyngbya_ANT.L52.2    <NA>
## 810                              Chamaesiphon_PCC-7430    <NA>
## 811                                 Calothrix_PCC-6303    <NA>
## 812                                 Wilmottia_Ant-Ph58    <NA>
## 813                               Microcoleus_PCC-7113    <NA>
## 814                                Phormidium_IAM_M-71    <NA>
## 815                             Synechococcus_PCC-7902    <NA>
## 816                                          Weissella    <NA>
## 817                                    Catenibacterium    <NA>
## 818                             Erysipelatoclostridium    <NA>
## 819                                          Breznakia    <NA>
## 820                                     Asteroleplasma    <NA>
## 821                                            UCG-004    <NA>
## 822                                         uncultured    <NA>
## 823                                          T2WK15B57    <NA>
## 824                                             NED5E9    <NA>
## 825                                       Anaeroplasma    <NA>
## 826                                       Entomoplasma    <NA>
## 827                                         Mesoplasma    <NA>
## 828                                        Spiroplasma    <NA>
## 829                                           RsaHf231    <NA>
## 830                                         Holdemania    <NA>
## 831                                         uncultured    <NA>
## 832                      [Anaerorhabdus]_furcosa_group    <NA>
## 833                            Candidatus_Hepatoplasma    <NA>
## 834                                Erysipelotrichaceae    <NA>
## 835                                     Faecalibaculum    <NA>
## 836                                         Dubosiella    <NA>
## 837                                       Faecalitalea    <NA>
## 838                             Candidatus_Spiroplasma    <NA>
## 839                                           type_III    <NA>
## 840                                          Oxobacter    <NA>
## 841                        [Eubacterium]_nodatum_group    <NA>
## 842                                      Mucispirillum    <NA>
## 843                                     Cloacibacillus    <NA>
## 844                                Candidatus_Tammella    <NA>
## 845                                   Anaeromyxobacter    <NA>
## 846                                      Paenibacillus    <NA>
## 847                                         uncultured    <NA>
## 848                                         Arenimonas    <NA>
## 849                                         Tahibacter    <NA>
## 850                                  Pseudoxanthomonas    <NA>
## 851                                      Rhodanobacter    <NA>
## 852                                   Stenotrophomonas    <NA>
## 853                                      Desulfovibrio    <NA>
## 854                                         uncultured    <NA>
## 855                                       Microvirgula    <NA>
## 856                                       Pusillimonas    <NA>
## 857                                            Deefgea    <NA>
## 858                                             mle1-7    <NA>
## 859                                          Ellin6067    <NA>
## 860                          Candidatus_Accumulibacter    <NA>
## 861                                        Bergeriella    <NA>
## 862                                              R7C24    <NA>
## 863                                      Acinetobacter    <NA>
## 864                                                T34    <NA>
## 865                                      endosymbionts    <NA>
## 866                                Candidatus_Annandia    <NA>
## 867                                           Buchnera    <NA>
## 868                                        Providencia    <NA>
## 869                                        Pasteurella    <NA>
## 870                                         Cellvibrio    <NA>
## 871                                             BIyi10    <NA>
## 872                                         uncultured    <NA>
## 873                                      Methylotenera    <NA>
## 874                                     Propionivibrio    <NA>
## 875                                             DSSD61    <NA>
## 876                                              966-1    <NA>
## 877                                              IS-44    <NA>
## 878                                         Izhakiella    <NA>
## 879                                        Actinospica    <NA>
## 880                                      Mycobacterium    <NA>
## 881                                       Anaerofustis    <NA>
## 882                                         uncultured    <NA>
## 883                                               cvE6    <NA>
## 884                                        Subgroup_25    <NA>
## 885                                Christensenellaceae    <NA>
## 886                                        Bacteroides    <NA>
## 887                             Prevotellaceae_UCG-004    <NA>
## 888                            Lachnospiraceae_UCG-004    <NA>
## 889                                  Lachnoclostridium    <NA>
## 890                                          Roseburia    <NA>
## 891                                       Dehalobacter    <NA>
## 892                                     Vicinamibacter    <NA>
## 893                                         uncultured    <NA>
## 894                                            UKL13-1    <NA>
## 895                                      Moraxellaceae    <NA>
## 896                                      Cetobacterium    <NA>
## 897                                      Simplicispira    <NA>
## 898                                        Variibacter    <NA>
## 899 Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium    <NA>
## 900                                             4-29-1    <NA>
## 901                                         uncultured    <NA>
## 902                                    Roseisolibacter    <NA>
## 903                                   Geodermatophilus    <NA>
## 904                                         uncultured    <NA>
## 905                                           AKYG1722    <NA>
## 906                                            Thermus    <NA>
## 907                                      Proteocatella    <NA>
## 908                                       Alkanindiges    <NA>
## 909                                       Turicibacter    <NA>
## 910                                          vadinBE97    <NA>
## 911                                          P.palmC41    <NA>
## 912                                             DTU089    <NA>
## 913                                      Rikenellaceae    <NA>
## 914                                          Alistipes    <NA>
## 915                                   dgA-11_gut_group    <NA>
## 916                                          Rikenella    <NA>
## 917                                       Mucinivorans    <NA>
## 918                                       Roseimarinus    <NA>
## 919                                   Negativibacillus    <NA>
## 920                                         uncultured    <NA>
## 921                                    Parabacteroides    <NA>
## 922                                         Tannerella    <NA>
## 923                                       Dysgonomonas    <NA>
## 924                                     Tannerellaceae    <NA>
## 925                                 Macellibacteroides    <NA>
## 926                               Rs-E47_termite_group    <NA>
## 927                              Bacteroidales_UCG-001    <NA>
## 928                                     Alloprevotella    <NA>
## 929                                     Proteiniphilum    <NA>
## 930                                        Thiodictyon    <NA>
## 931                                       Ruminococcus    <NA>
## 932                                           Bacillus    <NA>
## 933                                          0319-6G20    <NA>
## 934                                         Paracoccus    <NA>
## 935                        Clostridium_sensu_stricto_5    <NA>
## 936                                           FFCH5858    <NA>
## 937                           Bacteroidales_RF16_group    <NA>
## 938                     [Eubacterium]_ventriosum_group    <NA>
## 939                                  Sanguibacteroides    <NA>
## 940                                        Odoribacter    <NA>
## 941                                   Plasticicumulans    <NA>
## 942                      Lachnospiraceae_NK4A136_group    <NA>
## 943                                         uncultured    <NA>
## 944                                     Erysipelothrix    <NA>
## 945                              Phascolarctobacterium    <NA>
## 946                                         uncultured    <NA>
## 947                                       Succinispira    <NA>
## 948                            Candidatus_Stoquefichus    <NA>
## 949                       Lachnospiraceae_NC2004_group    <NA>
## 950                                      Robinsoniella    <NA>
## 951                    [Ruminococcus]_gauvreauii_group    <NA>
## 952                                         uncultured    <NA>
## 953                                             KD4-96    <NA>
## 954                                     Elusimicrobium    <NA>
## 955                                Gastranaerophilales    <NA>
## 956                                 Loriellopsis_LF-B5    <NA>
## 957                                         Tuzzerella    <NA>
## 958                                            CAG-352    <NA>
## 959                                     Chthoniobacter    <NA>
## 960                                        Akkermansia    <NA>

results_DESeq2$obj_significant

## class: TreeSummarizedExperiment 
## dim: 22 42 
## metadata(0):
## assays(1): counts
## rownames(22): OTU270 OTU333 ... OTU7070 OTU7691
## rowData names(7): Kingdom Phylum ... Genus Species
## colnames(42): UHM1240.20566_S41 UHM1248.20575_S54 ... UHM470.20533_S25
##   UHM476.20414_S46
## colData names(34): sample.id X16S.biosample ... swab.presence MK.spike
## reducedDimNames(0):
## mainExpName: NULL
## altExpNames(0):
## rowLinks: a LinkDataFrame (22 rows)
## rowTree: 1 phylo tree(s) (22 leaves)
## colLinks: NULL
## colTree: NULL

# Optional: Visualization
 results_DESeq2$bar_plot

Differentially abundant bacterial taxa between Desmognathus imitator and D. monticola are shown (adjusted p < 0.05). Purple bars indicate taxa enriched in D. imitator (positive LFC) and teal bars those enriched in D. monticola (negative LFC). Bar length denotes log fold change; lines show 1 SE. Distinct enrichment patterns reveal host-specific bacterial community differentiation.

Differential abundance (Multilayer Pairwise)

# DspikeIn: Differential Abundance Examples
# Using perform_and_visualize_DA() with multiple contrast scenarios

# 1. Single Contrast

res_single <- perform_and_visualize_DA(
  obj = tse_absolute,
  method = "DESeq2",
  group_var = "Diet",
  contrast = c("Insectivore", "Carnivore"),
  target_glom = "Genus")



# 2. Single Factor  All Pairwise Contrasts
col_df <- as.data.frame(colData(tse_absolute))
col_df$Host.taxon <- factor(make.names(as.character(col_df$Host.taxon)))
colData(tse_absolute)$Host.taxon <- col_df$Host.taxon

# Get unique DESeq2-compatible factor levels
host_levels <- levels(col_df$Host.taxon)
print(host_levels)

# contrast list
contrast_named <- list(
  Host.taxon = combn(host_levels, 2, simplify = FALSE))

# multiple pairwise contrasts
res_multi <- perform_and_visualize_DA(
  obj = tse_absolute,
  method = "DESeq2",
  group_var = "Host.taxon",
  contrast = contrast_named,
  target_glom = "Genus")



# 3. Single Factor  Selected Contrasts

contrast_list <- list(
  c("Insectivore", "Carnivore"),
  c("Omnivore", "Herbivore"))

res_selected <- perform_and_visualize_DA(
  obj = tse_absolute,
  method = "DESeq2",
  group_var = "Diet",
  contrast = contrast_list,
  global_fdr = TRUE)



# 4. Multiple Factors  Selected Contrasts

contrast_named <- list(
  Diet = list(
    c("Insectivore", "Carnivore"),
    c("Omnivore", "Carnivore") ),
  Animal.type = list(
    c("Frog", "Salamander") ))

res_multi_factor <- perform_and_visualize_DA(
  obj = tse_absolute,
  method = "DESeq2",
  contrast = contrast_named,
  target_glom = "Genus",
  significance_level = 0.01,
  global_fdr = TRUE)



# 5. Multiple Factors  all pairwise contrasts

colData(tse_absolute)$Host.taxon <- droplevels(factor(colData(tse_absolute)$Host.taxon))
colData(tse_absolute)$Habitat <- droplevels(factor(colData(tse_absolute)$Habitat))
host_levels <- levels(colData(tse_absolute)$Host.taxon)
habitat_levels <- levels(colData(tse_absolute)$Habitat)

# Define pairwise contrasts as a named list
contrast_named <- list(
  Host.taxon = combn(host_levels, 2, simplify = FALSE),
  Habitat = combn(habitat_levels, 2, simplify = FALSE))

# Define ComboGroup variable
colData(tse_absolute)$ComboGroup <- factor(interaction(
  colData(tse_absolute)$Animal.type,
  colData(tse_absolute)$Diet,
  drop = TRUE))



# --- Run multi-factor DESeq2 comparisons ---
results_multi_factor <- perform_and_visualize_DA(
  obj = tse_absolute,
  method = "DESeq2",
  contrast = contrast_named,
  target_glom = "Genus",
  significance_level = 0.01)

# --- Combined Group Interactions ---
colData(tse_absolute)$ComboGroup <- factor(interaction(
  colData(tse_absolute)$Animal.type,
  colData(tse_absolute)$Diet,
  drop = TRUE))

# --- Define custom interaction contrasts ---
contrast_list <- list(
  c("Salamander.Insectivore", "Lizard.Insectivore"),
  c("Salamander.Carnivore", "Snake.Carnivore"),
  c("Salamander.Carnivore", "Frog.Carnivore"))

# --- Run combined group comparisons ---
res_combo <- perform_and_visualize_DA(
  obj = tse_absolute,
  method = "DESeq2",
  group_var = "ComboGroup",
  contrast = contrast_list,
  target_glom = "Genus",
  global_fdr = TRUE)

Turnover (Presence/absence analysis)

taxonomic detection consistency between relative abundance (RA) and AA tables Presence/absence analysis to detect concordance between AA and RA profiles

Purpose: This analysis compares taxonomic detection consistency between RA and AA datasets. By converting both OTU tables to presence/absence matrices and intersecting shared taxa and samples, we assess how many taxa are consistently detected in both datasets.

Interpretation: The resulting “percent_shared” value represents the proportion of taxa jointly detected in RA and AA profiles, providing an estimate of detection concordance and turnover. Presenceabsence comparison between absolute (AA) and relative (RA) abundance datasets revealed that ~100% of taxa were shared, while the discrepancies likely reflect compositional bias introduced by relative normalization rather than true biological differences.

# --------------------------------------------
#  Extract OTU matrices from TSE objects
# --------------------------------------------
# relative dataset -> tse_desmog_rel
# tse_16SOTU_spiked_samples <- tse_16SOTU[, colData(tse_16SOTU)$spiked.volume %in% c("2", "1") ]

tse_filtered_rel <- tse_16SOTU_spiked_samples[
  rowData(tse_16SOTU_spiked_samples)$Genus != "Tetragenococcus" &
  rowData(tse_16SOTU_spiked_samples)$Family != "Chloroplast" &
  rowData(tse_16SOTU_spiked_samples)$Order != "Chloroplast", ]

tse_caudate_rel <- tse_filtered_rel[, colData(tse_filtered_rel)$Clade.Order == "Caudate"]
genera_keep <- c("Desmognathus", "Plethodon", "Eurycea")
tse_three_genera_rel <- tse_caudate[, colData(tse_caudate_rel)$Host.genus %in% genera_keep]
tse_blue_ridge_rel <- tse_three_genera_rel[, colData(tse_three_genera_rel)$Ecoregion.III == "Blue Ridge"]
tse_desmog_rel <- tse_blue_ridge_rel[, colData(tse_blue_ridge_rel)$Host.genus == "Desmognathus"]

# absolute dataset -> tse_desmog

otu_rel <- assay(tse_desmog_rel, "counts") 
otu_abs <- assay(tse_desmog, "counts")        

# Make sure samples are rows and taxa are columns
otu_rel <- t(otu_rel)
otu_abs <- t(otu_abs)

# Ensure they are matrices
otu_rel <- as.matrix(otu_rel)
otu_abs <- as.matrix(otu_abs)

# --------------------------------------------
# 2. Convert to Presence/Absence 
# --------------------------------------------
otu_rel_pa <- (otu_rel > 0) * 1
otu_abs_pa <- (otu_abs > 0) * 1

# --------------------------------------------
# 3. Identify common samples & taxa
# --------------------------------------------
shared_samples <- intersect(rownames(otu_rel_pa), rownames(otu_abs_pa))
shared_taxa <- intersect(colnames(otu_rel_pa), colnames(otu_abs_pa))

# Subset to common set
otu_rel_pa <- otu_rel_pa[shared_samples, shared_taxa]
otu_abs_pa <- otu_abs_pa[shared_samples, shared_taxa]

# --------------------------------------------
# 4. Compare presence/absence profiles
# --------------------------------------------
shared_pa <- (otu_rel_pa + otu_abs_pa) == 2
total_present <- (otu_rel_pa + otu_abs_pa) >= 1

# Calculate percent agreement (global)
percent_shared <- sum(shared_pa) / sum(total_present)

cat("Percent of shared taxa detections (AA vs RA):",
    round(100 * percent_shared, 1), "%\n")

## Percent of shared taxa detections (AA vs RA): 100 %

Visualization

#   Visualization of community composition

# Subset rows where Genus is not Tetragenococcus and not NA
keep_taxa <- !is.na(rowData(tse_absolute)$Genus) & rowData(tse_absolute)$Genus != "Tetragenococcus"
# Subset the TSE by keeping only those taxa (rows)
tse_filtered <- tse_absolute[keep_taxa, ]
# Exclude blanks
tse_filtered <- tse_filtered[, colData(tse_filtered)$sample.or.blank != "blank"]
# subset Salamander samples
tse_sal <- tse_filtered[, colData(tse_filtered)$Animal.type == "Salamander"]
Prok_OTU_sal <- tidy_phyloseq_tse(tse_sal)

TB<-taxa_barplot(
  Prok_OTU_sal,
  target_glom = "Family",
  custom_tax_names = NULL,
  normalize = TRUE,
  treatment_variable = "Habitat",
  abundance_type = "relative",
  x_angle = 15,
  fill_variable = "Family",
  facet_variable = "Diet",
  top_n_taxa = 10,
  palette = DspikeIn::color_palette$cool_MG,
  legend_size = 11,
  legend_columns = 1,
  x_scale = "free",
  xlab = NULL)

Microbial dynamics & NetWork comparision

To create a microbial co-occurrence network, you can refer to the SpiecEasi package available at: SpiecEasi GitHub Repository //github.com/zdk123/SpiecEasi The weight_Network() function provides an integrated framework for exploring and quantifying microbial interaction networks. It imports a pre-computed or user-supplied GraphML network, computes key topological descriptors (e.g., modularity, connectivity, transitivity, and path length), and visualizes the graph using a force-directed layout. Node colors are assigned according to modularity classes, while edge thickness and color reflect the magnitude and sign of associations, respectively, enabling intuitive interpretation of network structure. Together, these analyses highlight how community organization and interaction patterns change under different ecological contexts, for instance, after removing hubs or specific taxa (e.g., Basidiobolus) as we showed (doi.org/10.1093/ismejo/wraf150). The resulting visualization and metrics provide complementary insights into network complexity and modular structure, serving as a quantitative basis for assessing microbiome stability and cross-domain connectivity.

# 1. Initialization and loading NetWorks for Comparision


# library(SpiecEasi)
# library(ggnet)
# library(igraph)
library(DspikeIn)
library(tidyr)
library(dplyr)
library(ggpubr)
library(igraph)


# herp.Bas is a merged phyloseq object for both bacterial and fungal domains
# spiec.easi(herp.Bas, method='mb', lambda.min.ratio=1e-3, nlambda=250,pulsar.select=TRUE )

Complete <- DspikeIn::load_graphml("Complete.graphml")
NoBasid <- DspikeIn::load_graphml("NoBasid.graphml")
NoHubs <- DspikeIn::load_graphml("NoHubs.graphml")

# degree_network -> please note that needs full path of your graphml
result_kk <- DspikeIn::degree_network(load_graphml("Complete.graphml"),
  save_metrics = TRUE,
  layout_type = "stress")

result_kk$metrics

##   Nodes Edges Modularity   Density Transitivity Diameter Avg_Path_Length
## 1   308  1144  0.7355347 0.0241973    0.1789773 1.090862      0.09823294
##   Avg_Degree
## 1   7.428571

 result <- DspikeIn::weight_Network(graph_path = "Complete.graphml")
 result$plot

Burts Theory of Structural Holes (Ronald S. Burt, 1992, 2004) & Network Closure and Redundancy (1992, 2001) Cohesion and Trust & Brokerage and Network Advantage (Burt, 2005) The BrokerageClosure Tradeoff & Redundancy and Efficiency (20072010) Non-redundant Contacts and Efficiency & Structural Autonomy (Burt, 1997) Freedom through Brokerage

Ronald Burt (1992) proposed that in a network, some nodes occupy structural holes, gaps between otherwise disconnected groups or clusters. A node that bridges these holes plays a unique role: it connects information or resources between otherwise separate modules. Bridging role of Basidiobolus consistent with Burts Structural Hole Theory. The network illustrates taxon-level associations among microbial domains. Distinct bacterial and fungal subnetworks form modular clusters (colored by domain or module). Basidiobolus acts as a bridging node connecting otherwise disconnected clusters, a position corresponding to a structural hole in Burts theory (Burt, 1992). Such brokerage positions facilitate information and metabolite exchange across domains, enhancing functional redundancy and community stability. Nodes are sized by betweenness centrality, emphasizing the high structural influence of Basidiobolus in maintaining network cohesion under environmental stress. Moreover, Basidioboluss links are non-redundant; it connects functionally distinct taxa that otherwise have no direct edge. This makes it ecologically efficient in facilitating novel functional exchanges (e.g., horizontal gene transfer or cross-domain metabolic complementation).

Visualizes pairwise relationships between node-level metrics using a quadrant-based plot

node_level_metrics() & quadrant_plot() functions quantify and visualize node-level structural properties within microbial association networks. By integrating multiple centrality and connectivity metrics, it characterizes each nodes influence, redundancy (based on Bert’s theory), and modular affiliation, allowing comparisons of network architecture and community organization. The resulting plots and table provide complementary views of node prominence and connectivity patterns, serving as a foundation for interpreting ecological stability, functional redundancy, and cross-domain interactions.

Metric	Description
‘Node’	Node name (character format)
‘Degree’	Number of edges connected to the node
‘Strength’	Sum of edge weights connected to the node
‘Closeness’	Closeness centrality (normalized, based on shortest paths)
‘Betweenness’	Betweenness centrality (normalized, measures control over network flow)
‘EigenvectorCentrality’	Eigenvector centrality (importance based on connections to influential nodes)
‘PageRank’	PageRank score (importance based on incoming links)
‘Transitivity’	Local clustering coefficient (tendency of a node to form triangles)
‘Coreness’	Node’s coreness (from k-core decomposition)
‘Constraint’	Burt’s constraint (measures structural holes in a node’s ego network)
‘EffectiveSize’	Inverse of constraint (larger values = more non-redundant connections)
‘Redundancy’	Sum of constraint values of a node’s alters
‘Community’	Community assignment from Louvain clustering
‘Efficiency’	Global efficiency (average inverse shortest path length)
‘Local_Efficiency’	Local efficiency (subgraph efficiency for a node’s neighbors)
‘Within_Module_Connectivity’	Proportion of neighbors in the same community
‘Among_Module_Connectivity’	Proportion of neighbors in different communities

# Load required libraries
library(igraph)
library(dplyr)
library(tidyr)
library(ggplot2)
library(ggrepel)
ggplot2::margin()

## [1] 0points 0points 0points 0points

# 2.  Metrics Calculation

completeMetrics <- DspikeIn::node_level_metrics(Complete)
NoHubsMetrics <- DspikeIn::node_level_metrics(NoHubs)
NoBasidMetrics <- DspikeIn::node_level_metrics(NoBasid)


Complete_metrics <- completeMetrics$metrics
Nohub_metrics <- NoHubsMetrics$metrics
Nobasid_metrics <- NoBasidMetrics$metrics

Complete_metrics <- data.frame(lapply(Complete_metrics, as.character), stringsAsFactors = FALSE)
Nohub_metrics <- data.frame(lapply(Nohub_metrics, as.character), stringsAsFactors = FALSE)
Nobasid_metrics <- data.frame(lapply(Nobasid_metrics, as.character), stringsAsFactors = FALSE)


print(igraph::vcount(Complete)) # Number of nodes in Complete network

## [1] 308

print(igraph::ecount(Complete)) # Number of edges in Complete network

## [1] 1144

print(igraph::vcount(NoBasid))

## [1] 307

print(igraph::ecount(NoBasid))

## [1] 1187

print(igraph::vcount(NoHubs))

## [1] 286

print(igraph::ecount(NoHubs))

## [1] 916

metrics_scaled <- bind_rows(
  Complete_metrics %>% mutate(Network = "Complete"),
  Nohub_metrics %>% mutate(Network = "NoHubs"),
  Nobasid_metrics %>% mutate(Network = "NoBasid")
) %>%
  dplyr::mutate(dplyr::across(where(is.numeric), scale))

metrics_long_scaled <- metrics_scaled %>%
  tidyr::pivot_longer(cols = -c(Node, Network), names_to = "Metric", values_to = "Value")


# Ensure each dataset has a "Network" column before combining
completeMetrics$metrics <- completeMetrics$metrics %>% mutate(Network = "Complete Network")
NoHubsMetrics$metrics <- NoHubsMetrics$metrics %>% mutate(Network = "Network & Module Hubs Removed")
NoBasidMetrics$metrics <- NoBasidMetrics$metrics %>% mutate(Network = "Basidiobolus Subnetwork Removed")

# Combine All Data
combined_data <- bind_rows(
  completeMetrics$metrics,
  NoHubsMetrics$metrics,
  NoBasidMetrics$metrics
)


# Add Node Identifier if missing
if (!"Node" %in% colnames(combined_data)) {
  combined_data <- combined_data %>% mutate(Node = rownames(.))
}

# Convert `Network` into Factor
combined_data$Network <- factor(combined_data$Network, levels = c(
  "Complete Network",
  "Network & Module Hubs Removed",
  "Basidiobolus Subnetwork Removed"))

# Convert Data to Long Format
metrics_long <- combined_data %>%
  pivot_longer(
    cols = c("Redundancy", "Efficiency", "Betweenness"),
    names_to = "Metric", values_to = "Value")


# Define Custom Colors and Shapes
  network_colors <- c(
  "Complete Network" = "#F1E0C5",
  "Network & Module Hubs Removed" = "#D2A5A1",
  "Basidiobolus Subnetwork Removed" = "#B2C3A8")
  
network_shapes <- c(
  "Complete Network" = 21,
  "Network & Module Hubs Removed" = 22,
  "Basidiobolus Subnetwork Removed" = 23)

# Determine Top 30% of Nodes to Label/Optional
metrics_long <- metrics_long %>%
  group_by(Network, Metric) %>%
  mutate(Label = ifelse(rank(-Value, ties.method = "random") / n() <= 0.3, Node, NA))

# ?quadrant_plot() can creat plot for indivisual network
 plot4 <- quadrant_plot(completeMetrics$metrics, x_metric = "Among_Module_Connectivity", y_metric = "Degree")

 
 
# Customized comparision Plots
create_metric_plot <- function(metric_name, data, title) {
  data_filtered <- data %>% filter(Metric == metric_name)
  median_degree <- median(data_filtered$Degree, na.rm = TRUE)
  median_value <- median(data_filtered$Value, na.rm = TRUE)

  ggplot(data_filtered, aes(x = Degree, y = Value, fill = Network)) +
    geom_point(aes(shape = Network), size = 3, stroke = 1, color = "black") +
    geom_text_repel(aes(label = Label), size = 3, max.overlaps = 50) +
    scale_fill_manual(values = network_colors) +
    scale_shape_manual(values = network_shapes) +
    geom_vline(xintercept = median_degree, linetype = "dashed", color = "black", size = 1) +
    geom_hline(yintercept = median_value, linetype = "dashed", color = "black", size = 1) +
    labs(
      title = title,
      x = "Degree",
      y = metric_name,
      fill = "Network",
      shape = "Network" ) +
    theme_minimal() +
    theme(
      plot.title = element_text(
        hjust = 0.5, size = 16, face = "bold",
        margin = margin(t = 10, b = 20) # Moves the title downward
      ),
      axis.title = element_text(size = 14, face = "bold"),
      legend.position = "top",
      legend.title = element_text(size = 14, face = "bold"),
      legend.text = element_text(size = 12)  )
}


# Generate Plots
plot_redundancy <- create_metric_plot("Redundancy", metrics_long, "Redundancy vs. Degree Across Networks")
plot_efficiency <- create_metric_plot("Efficiency", metrics_long, "Efficiency vs. Degree Across Networks")
plot_betweenness <- create_metric_plot("Betweenness", metrics_long, "Betweenness vs. Degree Across Networks")

# Print Plots
 print(plot_betweenness)

Betweenness-degree relationships illustrate how node influence and connectivity shift after hub or Basidiobolus subnetwork removal, identifying taxa that maintain global connectivity and serve as potential ecological keystones.

Comparision of node-level metrics

Compares standardized node-level metrics across network configurations (e.g., complete vs. hub-removed). Boxplots and Wilcoxon tests illustrate how structural perturbations alter node influence and connectivity, offering insights into microbial network robustness and modular reorganization.

# 2.  Metrics Calculation

result_Complete <- DspikeIn::node_level_metrics(Complete)
result_NoHubs <- DspikeIn::node_level_metrics(NoHubs)
result_NoBasid <- DspikeIn::node_level_metrics(NoBasid)

Complete_metrics <- result_Complete$metrics
Nohub_metrics <- result_NoHubs$metrics
Nobasid_metrics <- result_NoBasid$metrics

Complete_metrics <- data.frame(lapply(Complete_metrics, as.character), stringsAsFactors = FALSE)
Nohub_metrics <- data.frame(lapply(Nohub_metrics, as.character), stringsAsFactors = FALSE)
Nobasid_metrics <- data.frame(lapply(Nobasid_metrics, as.character), stringsAsFactors = FALSE)


print(igraph::vcount(Complete)) # Number of nodes in Complete network

## [1] 308

print(igraph::ecount(Complete)) # Number of edges in Complete network

## [1] 1144

print(igraph::vcount(NoBasid))

## [1] 307

print(igraph::ecount(NoBasid))

## [1] 1187

print(igraph::vcount(NoHubs))

## [1] 286

print(igraph::ecount(NoHubs))

## [1] 916

metrics_scaled <- bind_rows(
  Complete_metrics %>% mutate(Network = "Complete"),
  Nohub_metrics %>% mutate(Network = "NoHubs"),
  Nobasid_metrics %>% mutate(Network = "NoBasid")
) %>%
  dplyr::mutate(dplyr::across(where(is.numeric), scale))

metrics_long_scaled <- metrics_scaled %>%
  tidyr::pivot_longer(cols = -c(Node, Network), names_to = "Metric", values_to = "Value")


# Remove missing values
metrics_long_scaled <- na.omit(metrics_long_scaled)

# We visualize only six metrics
selected_metrics <- c(
  "Degree", "Closeness", "Betweenness",
  "EigenvectorCentrality", "PageRank", "Transitivity"
)

metrics_long_filtered <- metrics_long_scaled %>%
  filter(Metric %in% selected_metrics) %>%
  mutate(
    Value = as.numeric(as.character(Value)),
    Network = recode(Network,
      "Complete" = "Complete Network",
      "NoHubs" = "Network & Module Hubs Removed",
      "NoBasid" = "Basidiobolus Subnetwork Removed" ) ) %>%
  na.omit() # Remove any NA if any

metrics_long_filtered$Network <- factor(metrics_long_filtered$Network,
  levels = c(
    "Complete Network",
    "Network & Module Hubs Removed",
    "Basidiobolus Subnetwork Removed" ))

# DspikeIn::color_palette$light_MG
network_colors <- c(
  "Complete Network" = "#F1E0C5",
  "Network & Module Hubs Removed" = "#D2A5A1",
  "Basidiobolus Subnetwork Removed" = "#B2C3A8")

# statistical comparisons a vs b
comparisons <- list(
  c("Complete Network", "Network & Module Hubs Removed"),
  c("Complete Network", "Basidiobolus Subnetwork Removed"),
  c("Network & Module Hubs Removed", "Basidiobolus Subnetwork Removed"))

networks_in_data <- unique(metrics_long_filtered$Network)
comparisons <- comparisons[sapply(comparisons, function(pair) all(pair %in% networks_in_data))]

met <- ggplot(metrics_long_filtered, aes(x = Network, y = Value, fill = Network)) +
  geom_boxplot(outlier.shape = NA) +
  geom_jitter(aes(color = Network),
    position = position_jitter(0.2), alpha = 0.2, size = 1.5 ) +
  scale_fill_manual(values = network_colors) +
  scale_color_manual(values = network_colors) +
  facet_wrap(~Metric, scales = "free_y", labeller = label_wrap_gen(width = 20)) +
  ggpubr::stat_compare_means(method = "wilcox.test", label = "p.signif", comparisons = comparisons) +
  theme_minimal() +
  theme(
    axis.title.x = element_blank(),
    axis.title.y = element_blank(),
    axis.text.x = element_text(size = 10, angle = 10, hjust = 0.8),
    strip.text = element_text(size = 12, margin = margin(t = 15, b = 5)),
    legend.position = "top",
    legend.text = element_text(size = 13),
    legend.title = element_text(size = 13, face = "bold"),
    plot.title = element_text(size = 13, face = "bold")
  ) + labs(title = "Selected Node Metrics Across Networks", fill = "Network Type", color = "Network Type")
met

Node-level topological metrics were compared across network configurations to assess structural robustness. Removal of the Basidiobolus subnetwork altered centrality measures, particularly closeness and PageRank, while maintaining overall network coherence.

To find neighbors of the target node

This function identifies both first- and second-degree neighbors of a given target node within a microbial association network. It enables users to trace direct and indirect ecological connections, revealing potential mediator taxa that bridge interactions between microbial guilds. The resulting summary table facilitates the inspection of a nodes local connectivity and extended influence zone, supporting hypothesis-driven exploration of cross-domain interactions and community structuring.

Complete <- load_graphml("Complete.graphml")
result2 <- DspikeIn::extract_neighbors(
  graph = Complete,
  target_node = "OTU69:Basidiobolus_sp", mode = "all")
head(result2$summary)

##             Type                           Node
## 1 First Neighbor            OTU8:Mortierella_sp
## 2 First Neighbor           OTU13:Mortierella_sp
## 3 First Neighbor           OTU15:Mortierella_sp
## 4 First Neighbor            OTU16:Ascomycota_sp
## 5 First Neighbor            OTU18:Helotiales_sp
## 6 First Neighbor OTU19:Margaritispora_monticola

Compute degree metrics and visualize the network

This analysis evaluates the global topology of microbial association networks under different structural configurationsnamely, the complete network, a hub-removed network, and a Basidiobolus-excluded subnetwork. Rather than inspecting individual nodes, this step focuses on overall network organization, including metrics that describe connectivity, efficiency, modularity, and cohesion.

The degree_network() function computes degree-based summaries and layouts for visualization, while weight_Network() extracts quantitative descriptors such as total edge weight, mean path length, and network density. Aggregating these metrics across network types enables direct comparison of topological robustness and information flow after targeted node or subnetwork removal.

The resulting bar plot displays log-scaled totals of each topological descriptor, allowing users to identify how network simplification (e.g., hub or guild removal) reshapes the overall architecture and resilience of the microbial network.

# Options: `"stress"` (default), `"graphopt"`, `"fr"`

# Compute degree metrics 
result <- degree_network(graph_path = Complete, save_metrics = TRUE)

# Compute network weights for different graph structures
NH <- weight_Network(graph_path = "NoHubs.graphml")
NB <- weight_Network(graph_path = "NoBasid.graphml")
C  <- weight_Network(graph_path = "Complete.graphml")

# Extract metrics from the computed network weights
CompleteM <- C$metrics
NoHubsM   <- NH$metrics
NoBasidM  <- NB$metrics

# Combine metrics into a single dataframe for comparison
df <- bind_rows(
  CompleteM %>% mutate(Group = "Complete Network"),
  NoHubsM %>% mutate(Group = "Network & Module Hubs Removed"),
  NoBasidM %>% mutate(Group = "Basidiobolus Subnetwork Removed")
) %>%
  pivot_longer(cols = -Group, names_to = "Metric", values_to = "Value")

# Aggregate total values by metric and group
df_bar <- df %>%
  group_by(Metric, Group) %>%
  summarise(Total_Value = sum(Value, na.rm = TRUE), .groups = "drop")

# Order metrics by mean value (optional)
metric_order <- df_bar %>%
  group_by(Metric) %>%
  summarise(mean_value = mean(Total_Value)) %>%
  arrange(desc(mean_value)) %>%
  pull(Metric)

df_bar$Metric <- factor(df_bar$Metric, levels = metric_order)

# Define refined color palette
network_colors <- c(
  "Complete Network" = "#F1E0C5",
  "Network & Module Hubs Removed" = "#D2A5A1",
  "Basidiobolus Subnetwork Removed" = "#B2C3A8"
)

# Create elegant bar plot
pg <- ggplot(df_bar, aes(x = Metric, y = log1p(Total_Value), fill = Group)) +
  geom_bar(stat = "identity", position = position_dodge(width = 0.8), width = 0.7, alpha = 0.9) +
  scale_fill_manual(values = network_colors) +
  theme_minimal(base_size = 14) +
  theme(
    panel.grid.major.x = element_blank(),
    panel.grid.minor = element_blank(),
    axis.text.x = element_text(size = 11, angle = 15, hjust = 1, vjust = 1, color = "gray20"),
    axis.text.y = element_text(size = 11, color = "gray25"),
    axis.title = element_text(size = 13, face = "bold"),
    plot.title = element_text(size = 15, face = "bold", hjust = 0.5, margin = ggplot2::margin(b = 10)),
    legend.position = "top",
    legend.title = element_blank(),
    legend.text = element_text(size = 12),
    panel.background = element_rect(fill = "white", color = NA),
    plot.background = element_rect(fill = "white", color = NA)
  ) +
  labs(
    subtitle = "Comparison of total network-level metrics (log-scaled)",
    x = NULL,
    y = "Total Value (log1p)"
  ) +
  geom_text(
    aes(label = round(log1p(Total_Value), 1)),
    position = position_dodge(width = 0.8),
    vjust = -0.3,
    size = 3.5,
    color = "gray25"
  )

Small-World Index Analysis

To determine whether the complete network exhibited small-world topology, we computed the Small-World Index (SWI; ) following the quantitative framework established by Humphries M, Gurney K, 2008. PLoS ONE 4 (Eq.1) (SWI; ) = (Global clustering coefficient real/Global clustering coefficient random)/(Avg Path real/Avg Path random))

library(igraph)
library(tidygraph)
library(ggraph)
library(DspikeIn)


# AA abundance
Complete<-load_graphml("Complete.graphml")

deg <- degree(Complete)

hist(deg, breaks = 30, main = "Degree Distribution", xlab = "Degree")
fit <- fit_power_law(deg + 1)  # Avoid zero degrees
print(fit)


# ---- empirical network ----
g_empirical = Complete
#g_empirical = Complete_Rel



# ---- Calculate real network metrics ----
creal <- transitivity(g_empirical, type = "global")
E(g_empirical)$weight <- abs(E(g_empirical)$weight)
lreal <- mean_distance(g_empirical, directed = FALSE, unconnected = TRUE, weights = E(g_empirical)$weight)

# ---- Generate 1000 Erds-Rnyi random graphs with same size ----
set.seed(42)  # for reproducibility
n_nodes <- vcount(g_empirical)
n_edges <- ecount(g_empirical)

crand_vals <- numeric(1000)
lrand_vals <- numeric(1000)

for (i in 1:1000) {
g_rand <- sample_gnm(n = n_nodes, m = n_edges, directed = FALSE)
if (!is_connected(g_rand)) next
  
  crand_vals[i] <- transitivity(g_rand, type = "global")
  lrand_vals[i] <- mean_distance(g_rand, directed = FALSE, unconnected = TRUE)
}

# ---- Calculate mean values across random graphs ----
crand <- mean(crand_vals, na.rm = TRUE)
lrand <- mean(lrand_vals, na.rm = TRUE)

# ---- Compute Small-World Index () ----
sigma <- (creal / crand) / (lreal / lrand)


cat("Global clustering coefficient (real):", creal, "\n")
cat("Average path length (real):", lreal, "\n")
cat("Mean clustering coefficient (random):", crand, "\n")
cat("Mean path length (random):", lrand, "\n")
cat("Small-World Index ():", round(sigma, 2), "\n")

= 231.83 is extremely high, showing very strong small-world properties. > 1 indicates a small-world network,combining high clustering with short paths.

sessionInfo()

## R version 4.5.2 (2025-10-31)
## Platform: x86_64-pc-linux-gnu
## Running under: Ubuntu 24.04.3 LTS
## 
## Matrix products: default
## BLAS:   /usr/lib/x86_64-linux-gnu/openblas-pthread/libblas.so.3 
## LAPACK: /usr/lib/x86_64-linux-gnu/openblas-pthread/libopenblasp-r0.3.26.so;  LAPACK version 3.12.0
## 
## locale:
##  [1] LC_CTYPE=en_US.UTF-8       LC_NUMERIC=C              
##  [3] LC_TIME=en_US.UTF-8        LC_COLLATE=C              
##  [5] LC_MONETARY=en_US.UTF-8    LC_MESSAGES=en_US.UTF-8   
##  [7] LC_PAPER=en_US.UTF-8       LC_NAME=C                 
##  [9] LC_ADDRESS=C               LC_TELEPHONE=C            
## [11] LC_MEASUREMENT=en_US.UTF-8 LC_IDENTIFICATION=C       
## 
## time zone: Etc/UTC
## tzcode source: system (glibc)
## 
## attached base packages:
## [1] stats4    stats     graphics  grDevices utils     datasets  methods  
## [8] base     
## 
## other attached packages:
##  [1] ggrepel_0.9.6                   igraph_2.2.1                   
##  [3] ggpubr_0.6.2                    tidyr_1.3.1                    
##  [5] vegan_2.7-2                     permute_0.9-8                  
##  [7] microbiome_1.32.0               tibble_3.3.0                   
##  [9] mia_1.18.0                      MultiAssayExperiment_1.36.0    
## [11] dplyr_1.1.4                     flextable_0.9.10               
## [13] ggplot2_4.0.0                   ggstar_1.0.6                   
## [15] TreeSummarizedExperiment_2.18.0 SingleCellExperiment_1.32.0    
## [17] Biostrings_2.78.0               XVector_0.50.0                 
## [19] phyloseq_1.54.0                 DspikeIn_1.0.0                 
## [21] SummarizedExperiment_1.40.0     Biobase_2.70.0                 
## [23] GenomicRanges_1.62.0            Seqinfo_1.0.0                  
## [25] IRanges_2.44.0                  S4Vectors_0.48.0               
## [27] BiocGenerics_0.56.0             generics_0.1.4                 
## [29] MatrixGenerics_1.22.0           matrixStats_1.5.0              
## [31] rmarkdown_2.30                 
## 
## loaded via a namespace (and not attached):
##   [1] ggtext_0.1.2                fs_1.6.6                   
##   [3] DirichletMultinomial_1.52.0 RColorBrewer_1.1-3         
##   [5] tools_4.5.2                 backports_1.5.0            
##   [7] R6_2.6.1                    lazyeval_0.2.2             
##   [9] mgcv_1.9-4                  rhdf5filters_1.22.0        
##  [11] withr_3.0.2                 gridExtra_2.3              
##  [13] cli_3.6.5                   textshaping_1.0.4          
##  [15] officer_0.7.0               slam_0.1-55                
##  [17] labeling_0.4.3              sass_0.4.10                
##  [19] mvtnorm_1.3-3               S7_0.2.0                   
##  [21] readr_2.1.5                 randomForest_4.7-1.2       
##  [23] ggridges_0.5.7              askpass_1.2.1              
##  [25] systemfonts_1.3.1           yulab.utils_0.2.1          
##  [27] scater_1.38.0               decontam_1.30.0            
##  [29] parallelly_1.45.1           limma_3.66.0               
##  [31] readxl_1.4.5                fillpattern_1.0.2          
##  [33] gridGraphics_0.5-1          car_3.1-3                  
##  [35] zip_2.3.3                   rbiom_2.2.1                
##  [37] Matrix_1.7-4                biomformat_1.38.0          
##  [39] ggbeeswarm_0.7.2            DECIPHER_3.6.0             
##  [41] abind_1.4-8                 lifecycle_1.0.4            
##  [43] yaml_2.3.10                 edgeR_4.8.0                
##  [45] carData_3.0-5               rhdf5_2.54.0               
##  [47] SparseArray_1.10.1          Rtsne_0.17                 
##  [49] grid_4.5.2                  crayon_1.5.3               
##  [51] lattice_0.22-7              beachmat_2.26.0            
##  [53] sys_3.4.3                   maketools_1.3.2            
##  [55] pillar_1.11.1               knitr_1.50                 
##  [57] estimability_1.5.1          codetools_0.2-20           
##  [59] fastmatch_1.1-6             glue_1.8.0                 
##  [61] ggiraph_0.9.2               ggfun_0.2.0                
##  [63] fontLiberation_0.1.0        data.table_1.17.8          
##  [65] vctrs_0.6.5                 treeio_1.34.0              
##  [67] cellranger_1.1.0            gtable_0.3.6               
##  [69] cachem_1.1.0                xfun_0.54                  
##  [71] S4Arrays_1.10.0             tidygraph_1.3.1            
##  [73] coda_0.19-4.1               survival_3.8-3             
##  [75] iterators_1.0.14            statmod_1.5.1              
##  [77] bluster_1.20.0              nlme_3.1-168               
##  [79] ggtree_4.0.1                fontquiver_0.2.1           
##  [81] bslib_0.9.0                 irlba_2.3.5.1              
##  [83] vipor_0.4.7                 DBI_1.2.3                  
##  [85] ade4_1.7-23                 phangorn_2.12.1            
##  [87] DESeq2_1.50.1               tidyselect_1.2.1           
##  [89] emmeans_2.0.0               compiler_4.5.2             
##  [91] BiocNeighbors_2.4.0         xml2_1.4.1                 
##  [93] fontBitstreamVera_0.1.1     DelayedArray_0.36.0        
##  [95] scales_1.4.0                quadprog_1.5-8             
##  [97] rappdirs_0.3.3              stringr_1.6.0              
##  [99] digest_0.6.37               htmltools_0.5.8.1          
## [101] pkgconfig_2.0.3             sparseMatrixStats_1.22.0   
## [103] fastmap_1.2.0               rlang_1.1.6                
## [105] htmlwidgets_1.6.4           DelayedMatrixStats_1.32.0  
## [107] farver_2.1.2                jquerylib_0.1.4            
## [109] jsonlite_2.0.0              BiocParallel_1.44.0        
## [111] BiocSingular_1.26.0         magrittr_2.0.4             
## [113] polynom_1.4-1               Formula_1.2-5              
## [115] scuttle_1.20.0              ggplotify_0.1.3            
## [117] patchwork_1.3.2             Rhdf5lib_1.32.0            
## [119] Rcpp_1.1.0                  ape_5.8-1                  
## [121] ggnewscale_0.5.2            viridis_0.6.5              
## [123] gdtools_0.4.4               stringi_1.8.7              
## [125] ggalluvial_0.12.5           ggraph_2.2.2               
## [127] MASS_7.3-65                 plyr_1.8.9                 
## [129] parallel_4.5.2              graphlayouts_1.2.2         
## [131] splines_4.5.2               gridtext_0.1.5             
## [133] multtest_2.66.0             hms_1.1.4                  
## [135] msa_1.42.0                  locfit_1.5-9.12            
## [137] uuid_1.2-1                  ggtreeExtra_1.20.0         
## [139] ggsignif_0.6.4              buildtools_1.0.0           
## [141] reshape2_1.4.4              ScaledMatrix_1.18.0        
## [143] evaluate_1.0.5              tzdb_0.5.0                 
## [145] foreach_1.5.2               tweenr_2.0.3               
## [147] openssl_2.3.4               purrr_1.2.0                
## [149] polyclip_1.10-7             BiocBaseUtils_1.12.0       
## [151] ggforce_0.5.0               rsvd_1.0.5                 
## [153] broom_1.0.10                xtable_1.8-4               
## [155] tidytree_0.4.6              rstatix_0.7.3              
## [157] viridisLite_0.4.2           ragg_1.5.0                 
## [159] aplot_0.2.9                 memoise_2.0.1              
## [161] beeswarm_0.4.0              cluster_2.1.8.1

DspikeIn volume protocol

Spike-in volume Protocol;

The species Tetragenococcus halophilus (bacterial spike; ATCC33315) and Dekkera bruxellensis (fungal spike; WLP4642-White Labs) were selected as taxa to spike into gut microbiome samples as they were not found in an extensive collection of wildlife skin (GenBank BioProjects: PRJNA1114724, PRJNA 1114659) or gut microbiome samples. Stock cell suspensions of both microbes were grown in either static tryptic soy broth (T. halophilus) or potato dextrose broth (D. bruxellensis) for 72 hours then serially diluted and optical density (OD600) determined on a ClarioStar plate reader. Cell suspensions with an optical density of 1.0, 0.1, 0.01, 0.001 were DNA extracted using the Qiagen DNeasy Powersoil Pro Kit. These DNA isolations were used as standards to determine the proper spike in volume of cells to represent 0.1-10% of a sample (Rao et al., 2021b) Fecal pellets (3.1 1.6 mg; range = 1 5.1 mg) from an ongoing live animal study using wood frogs (Lithobates sylvaticus) were used to standardize the input material for the development of this protocol. A total of (n=9) samples were used to validate the spike in protocol. Each fecal sample was homogenized in 1mL of sterile molecular grade water then 250uL of fecal slurry was DNA extracted as above with and without spiked cells. Two approaches were used to evaluate the target spike-in of 0.1-10%, the range of effective spike-in percentage described in (Rao et al., 2021b), including 1) an expected increase of qPCR cycle threshold (Ct) value that is proportional to the amount of spiked cells and 2) the expected increase in copy number of T. halophilus and D. bruxellensis in spiked vs. unspiked samples. A standard curve was generated using a synthetic fragment of DNA for the 16S-V4 rRNA and ITS1 rDNA regions of T. halophilus and D. bruxellensis, respectively. The standard curve was used to convert Ct values into log copy number for statistical analyses (detailed approach in[2, 3]) using the formula y = -0.2426x + 10.584 for T. halophilus and y = -0.3071x + 10.349 for D. bruxellensis, where x is the average Ct for each unknown sample. Quantitative PCR (qPCR) was used to compare known copy numbers from synthetic DNA sequences of T. halophilus and D. bruxellensis to DNA extractions of T. halophilus and D. bruxellensis independently, and wood frog fecal samples with and without spiked cells. SYBR qPCR assays were run at 20ul total volume including 10ul 2X Quantabio PerfeCTa SYBR Green Fastmix, 1ul of 10uM forward and reverse primers, 1ul of ArcticEnzymes dsDNAse master mix clean up kit, and either 1ul of DNA for D. bruxellensis or 3ul for T. halophilus. Different volumes of DNA were chosen for amplification of bacteria and fungi due to previous optimization of library preparation and sequencing steps [3]. The 515F [4] and 806R [5] primers were chosen to amplify bacteria and ITS1FI2 [6] and ITS2 for fungi, as these are the same primers used during amplicon library preparation and sequencing. Cycling conditions on an Agilent AriaMX consisted of 95 C for 3 mins followed by 40 cycles of 95 C for 15 sec, 60 C for 30 sec and 72 C for 30 sec. Following amplification, a melt curve was generated under the following conditions including 95 C for 30 sec, and a melt from 60 C to 90 C increasing in resolution of 0.5 C in increments of a 5 sec soak time. To validate the spike in protocol we selected two sets of fecal samples including 360 samples from a diverse species pool of frogs, lizards, salamanders and snakes and a more targeted approach of 122 fecal samples from three genera of salamanders from the Plethodontidae. (Supplemental Table #). FFecal samples were not weighed in the field, rather, a complete fecal pellet was diluted in an equal volume of sterile water and standardized volume of fecal slurry (250L) extracted for independent samples.. A volume of 1ul T. halophilus (1847 copies) and 1ul D. bruxellensis (733 copies) were spiked into each fecal sample then DNA was extracted as above, libraries constructed, and amplicon sequenced on an Illumina MiSeq as in [7] .

<github.com/mghotbi/DspikeIn>

DspikeIn with TSE