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ABSTRACT

TRACING HYDROCARBON CONTAMINATION THROUGH HYPERALKALINE ENVIRONMENTS IN THE CALUMET REGION OF SOUTHEASTERN CHICAGO

Kathryn Quesnell, MS Department of Geology and Environmental Geosciences Northern Illinois University, 2016 Melissa Lenczewski, Director

The Calumet region of Southeastern Chicago was once known for industrialization, which left pollution as its legacy. Disposal of slag and other industrial wastes occurred in nearby wetlands in attempt to create areas suitable for future development. The waste creates an unpredictable, heterogeneous geology and a unique hyperalkaline environment. Upgradient to the field site is a former coking facility, where coke, creosote, and coal weather openly on the ground. Hydrocarbons weather into characteristic polycyclic aromatic hydrocarbons (PAHs), which can be used to create a fingerprint and correlate them to their original parent compound.

This investigation identified PAHs present in the nearby surface and groundwaters through use of gas chromatography/mass spectrometry (GC/MS), as well as investigated the relationship between the alkaline environment and the organic contamination. PAH ratio analysis suggests that the organic contamination is not mobile in the groundwater, and instead originated from the air. 16S rDNA profiling suggests that some microbial communities are influenced more by pH, and some are influenced more by the hydrocarbon pollution. BIOLOG Ecoplates revealed that most communities have the ability to metabolize ring structures similar to the shape of PAHs.

Analysis with bioinformatics using PICRUSt demonstrates that each community has microbes thought to be capable of hydrocarbon utilization. The field site, as well as nearby areas, are targets for habitat remediation and recreational development. In order for these remediation efforts to be successful, it is vital to understand the geochemistry, weathering, microbiology, and distribution of known contaminants. Many PAHs are toxic in both the environment and to humans, and are known to be both carcinogenic as well as bioaccumulate. Thus, understanding the relationship of PAHs to the geochemistry of this field site provides critical information for ongoing research and future remediation efforts.

NORTHERN ILLINOIS UNIVERSITY DE KALB, ILLINOIS

AUGUST 2016

TRACING HYDROCARBON CONTAMINATION THROUGH HYPERALKALINE

ENVIRONMENTS IN THE CALUMET REGION OF SOUTHEASTERN CHICAGO

A THESIS SUBMITTED TO THE GRADUATE SCHOOL

IN PARTIAL FULFILLMENT OF THE REQUIREMENTS

FOR THE DEGREE

MASTER OF SCIENCE

DEPARTMENT OF GEOLOGY AND ENVIRONMENTAL GEOSCIENCES

Thesis Director: Melissa E. Lenczewski

ACKNOWLEDGEMENTS

There are far too many people to thank for the creation of this project.

For their aid in funding this project, I would like to thank the Illinois Groundwater

Association and the Geological Society of America for their generous support. From Northern

Illinois University, I would like to thank NIU Department of Geology and Environmental

Geosciences’ Samuel Goldich Fund, The NIU Graduate School’s Travel Fund and the College of

Liberal Arts and Sciences for further funding my research. For funding me, personally, I would like to thank the family of Brian Fugiel for their generous scholarship through the department, as well as The Institute for the Study of the Environment, Sustainability, and Energy/Environmental

Studies. For his generous donation of time and lab analysis, I would like to thank Stan Zaworski at First Environmental Laboratories in Naperville, Illinois.

For laboratory support, I would like to thank Monica Carroll, Ellen Raimondi, and the

Swingley Lab.

Finally, I would like to thank my thesis committee, Melissa Lenczewski, Wes Swingley, and Philip Carpenter for their patience and understanding in the face of every small project- related emergency.

DEDICATION

To my parents, for supporting me, and, more importantly, for never questioning me on this crazy journey.

TABLE OF CONTENTS

LIST OF TABLES………………………………………………..…………...………...... …vi

LIST OF FIGURES…………………………………………………………………….…….....vii

LIST OF APPENDICES……………………..……………………………………………….…viii

Chapter

1. INTRODUCTION ...... 1

2. BACKGROUND ...... 3

History of the Calumet Region……………………………………………………………3

Modern Field Site…………………………………………………………………………5

PAHS and the Environment……………………………………………………………….7

Alkaline Environments…………………………………………………………………..10

Field Methods of Investigation…………………………………………………………..11

3. MATERIALS AND METHODS ...... 13

Sampling Periods……………………………………………...…………………………13

Field Sampling Methods…………………………………………………………………13

Organic Chemistry: PAH Analysis……………………………………………………….15

Aqueous Geochemistry…………………………………………………………………..15

Biological Filtration and 16S rDNA Sequencing…………………………………………16

BIOLOG Ecoplates………………………………………………………………………17

4. DATA ANALYSIS AND DISCUSSION……………………………………………………..19

Inorganic Chemistry……...………………………………………………………………19 v

Organic Geochemistry…………………………………………………………………...21

Biology…………………………………………………………………………………...28

5. CONCLUSION………………………………...……………………………………………...42

REFERENCES…………………………………………………………………………………..44

APPENDICES…………………………………………………………………………………...50

LIST OF TABLES

Page Table 2.1 16 EPA Priority Pollutants……...………………………………………….……………9 Table 3.1 Collected Field Samples Summary……………...... …………………….………….14 Table 4.1 Summer Water Sample Inorganic Characterization (8/18/2014)…………..……...... …19 Table 4.2 Winter Water Sample Inorganic Characterization (12/16/2014)…………...... ………21 Table 4.3 Summer 16 EPA PAH Priority Pollutant Data (8/18/2014)…………………..……...... 22 Table 4.4 Winter 16 EPA PAH Priority Pollutant Data (12/16/2014)……………………..……...23 Table 4.5 Summer PAH Pearson Coefficients (12/16/2014)…….……………………………….24 Table 4.6 Winter PAH Pearson Coefficients (8/18/2014)………………………………………...24 Table 4.7 16 EPA Priority Pollutant Mobility…………………………………………………….28 Table 4.8 Structure of BIOLOG Carbon Substrates………………………………………………29 Table 4.9 Total PAHS of Selected Sites………………………………………………………….33 Table 4.10 Diversity Indices (from Class Data)…………………………………………………..38 Table 4.11 PICRUSt PAH KEGG Pathways One-Sample T-test………………………………...40

LIST OF FIGURES

Page Figure 2.1 Overview of the Area of Investigation…………………………………………………5 Figure 2.2 Overview of Big Marsh Transect……………….....……………………………………6 Figure 2.3 Overview of the Slag Dunes and Wolf Lake Comparison Site ………………..……...... 7 Figure 4.1 FL/PY to PH/AN Ratios of Select Calumet Samples Compared to Known Sources...... 26 Figure 4.2 BIOLOG Total Microbial Growth……………………………………………………30 Figure 4.3 pH Related to Microbial Growth……………………………………………………...31 Figure 4.4 BIOLOG PCA Data…………………………………………………………………...32 Figure 4.5 BIOLOG Carbon Structure Utilization………………………………………………..33 Figure 4.6 Summer 16S rDNA Microbial Profiles by Class……………………………………...34 Figure 4.7 16S rDNA Profiles PCA………………………………………………………………37 Figure 4.8 PAH-Related KEGG Pathways………………………………………………….……39 Figure 4.9 Hydrocarbon Degradation KEGG Pathways Related to Total PAHs………………….41

LIST OF APPENDICES

Appendix Page

A. 16S RDNA DATA…………………………………………………...………………………..50 B. 16S RDNA PCA DATA……………………………………………..………………………113 C. PCA DATA BIOLOGS……………………………………………………………………..116 D. PICRUST DATA……………………………………………………………………………119 E. BIOLOG DATA……………………………...……………………………………………..139

CHAPTER 1

INTRODUCTION

The Calumet region of Southeastern Chicago archives its industrial legacy in pollution.

Slag waste and garbage were used to fill marshes to make land suitable for residential and industrial development (Colton, 1986). Previous research of the area explored chemical reactions caused by the interaction of meteoric water and slag waste, creating the hyperalkaline conditions up to pH 13.4 of the ground and surface waters of the area (Roadcap et al., 2005; Roadcap et al., 2006). Immediately surrounding the hyperalkaline waters is an Army Corps of Engineers habitat restoration project, where species such as the Black Crowned Night Heron (endangered in the state of Illinois) live. The field site itself is being converted by the Chicago Park District into bike trails. Landfills in the area formerly produced leachate springs near the area of investigation

(Waska, 2014). The former property of Acme Steel (later Acme Coke) has coke, creosote, and coal lying openly and weathering on the ground.

Waska (2014) focused on characterization of groundwater flow paths, site-dependent microbial community characterization based on geochemistry, and hyperalkaline water remediation methods. However, in his investigation, he also noted unknown organic contamination present in both surface and groundwaters, but did not pursue investigation of the contamination further. 2

The purpose of this research is to characterize the organic contamination discovered in previous studies, and determine if the contamination is linked to the exposed hydrocarbons on the former Acme Coke Facility. Additionally, the microbial communities and their relationship with the organic contamination will be investigated.

CHAPTER 2

BACKGROUND

History of the Calumet Region

The natural surface and near-surface geology of the Calumet region is the result of previous glaciation. Although the area was believed to have 20-30 feet of Illinoian Stage drift, most, if not all, of the drift in the region has been removed by the Wisconsin Stage glaciation.

The Calumet area itself lies primarily on the former Lake Chicago basin, deposited approximately 12,500-75,000 years ago during the Equality Formation and consists of sand, silt, clay, and gravel deposits. The glacial deposits overly Silurian-aged dolostone bedrock (Willman,

1971).

In the 1870’s, Congress approved plans to dredge, widen, channelize, and straighten the

Calumet River for the purposes of industrializing the area (Sellers, 2006). Development of the

Calumet River was spurred by convenience— the nearby Chicago River was already crowded.

(Colten, 1986). Industrialization permitted the development of what would later become Acme

Steel/Coke in 1905. After multiple mergers and name changes, in 1984 the field site became known as Acme Steel. In 2001, after bankruptcy, Acme Steel was closed and the last blast furnace was finally demolished in 2004 (Sellers, 2006).

Sediment filling of the wetlands began in 1869 when the Calumet River was being widened (Colten, 1986). Waste disposal was largely unregulated. Industries along the river were 4 free to dispose of waste including heavy metals, organic pollution, steel quenching liquids, and solid wastes in any manner. Solid wastes were openly dumped until the 1940s in attempt to reclaim the land, with some industrial sites adding hundreds of acres to their property in this manner. When there was regulation, it largely focused on preventing the spread of infectious diseases from sewage and biohazardous wastes (Colten, 1986).

In the midst of industrial development, middle and lower class housing developments were created. This includes areas such as Altgeld Gardens, a government housing project, but also middle class neighborhoods such as Hegewisch. In addition to industrial emissions, the

Calumet area is surrounded by at least 21 different landfills (Bouman, 2001), some of which have been found to be leaky (Waska, 2014).

The slag fill and waste in this area originates from the manufacture of steel, and while it varies in composition it is in large part calcium-silicates, forming minerals such as Rankinite

(퐶푎3푆𝑖2푂7), Larnite (퐶푎2푆𝑖푂4), and Akerminite (퐶푎2푀푔푆𝑖2푂7). When these minerals interact with groundwater, they release both hydroxides and calcium ions. The hydroxides create hyperalkaline conditions, varying depending on temperature. When the groundwater reaches the surface, the free calcium ions interact with the carbon dioxide, and calcium carbonate is precipitated, forming thick, loose, white sediment (Roadcap et al., 2005).

Due to the heterogeneous nature of the fill, the geochemistry of the study sites varies.

However, previous studies have documented heavy metal and organic contamination (Waska,

2014; Roadcap and Kelly, 1996), the unique microbial communities (Roadcap et al., 2006), as well the potential options for remediation (Waska, 2014).

Modern Field Site

The main area of investigation is Big Marsh (figure 2.1), an area of hyperalkaline surface and groundwaters (Waska, 2014). This site is immediately downgradient from the former Acme

Coking Facility (Waska, 2014).

Overview of the Area of Investigation

Figure 2.1 Overview of the area of investigation. The former coking plant (A) is shown adjacent to the Army Corps of Engineer restoration area (B) and near formerly leaking landfills (C). Big Marsh, an area with hyperalkaline, yellow-colored surface water (D), has calcium carbonate precipitation visible from satellite images. Near Wolf Lake (E) is the control well located at the slag dunes (F). Pre-existing monitoring wells are seen in the slag (X) (Google Maps, 2014).

The area known as “Creosote Marsh” is a wetland area that is on the grounds of the former Acme Coking facility, on top of discarded coke, creosote, and coal. The RR Ditch runs 6 north/south, bisecting the transect and separating the coking facility from Big Marsh. Samples were taken from a ditch on the western side of the tracks (figure 2.2)

Overview of Big Marsh Transect

Figure 2.2: Detailed view of transect (see the area surround X in figure 2.1). Site #2 Monitoring well (A) is in between the creosote spring (B) and a Railroad Ditch (C). The banks of the Creosote Marsh (D) are saturated creosote, and are where solid creosote was obtained.

Wolf Lake (figure 2.3) is a recreational area within William Powers State Park, has a near-neutral pH (Waska, 2014), but also lies on slag fill (Roadcap et al., 2006). The Slag Dune area, immediately west of Wolf Lake, consists of piles of discarded slag. Groundwater and surface water from the small canal and the Slag Dunes are both hyperalkaline, and are not thought to be hydrologically connected to Big Marsh.

Overview of the Slag Dunes and Wolf Lake Comparison Site

Figure 2.3: Slag Dunes Field Site. Wolf Lake Lies to the East, and three monitoring wells line a small, hyperalkaline channel.

PAHs and the Environment

PAHs (Polycyclic Aromatic Hydrocarbons) are a common degradation product of coal, creosote, and coke (Neff et al., 2005). They are prevalent in urban environments, and can also result from automobile exhaust, wood burning, and factory emissions (Boehm, 2006). While there are several methods to degrade PAHs, in the environment they are primarily broken down by microbes. PAHs, while they provide energy sources for microbial life, (Haritash and Kaushik,

2009), are toxic to both humans and the environment (Toyooka and Ibuki, 2007). Exposure to certain PAHs has been shown to be carcinogenic to humans as well as having the ability to negatively impact the function of lung, kidney, and liver tissue (Samata et al., 2002). Research also suggests that PAHs can cause additional damage when exposed to UV radiation. Exposure 8 to UV radiation has been shown to increase the solubility of PAHs, increasing their availability throughout the water column (Davenport and Spacey, 2001). Organisms that have ingested PAHs and are later exposed to UV light have been shown to weaken the bonds of PAH molecules inside of them, creating oxygen free radicals (McDonald and Chapman, 2002). Previous investigations have suggested that these free radicals can damage cell membranes and mutate

DNA (Toyooka and Ibuki, 2007).

PAHs have low solubilities in water (Joa et al., 2009), but are lipophilic and therefore bioaccumulate in the ecosystem (Haritash and Kaushik, 2009). Many PAHs are suspected or known carcinogens (Haritash and Kaushik, 2009), and thus 16 PAHs (table 2.1) were included on the EPA’s Priority Pollutant List to be regulated by the Clean Water Act (EPA, 2014). The

EPA Priority Pollutant List was developed alongside the EPA Toxic Pollutant List (EPA, 2016).

The Toxic Pollutant list includes large groups of compounds, which cannot all be practically tested for in every investigation. Thus, the Priority Pollutant List is an attempt to represent the

Toxic Pollutant List (EPA, 2016). Recently, there has been discussion about revising the EPA

Priority Pollutant list. The list, while it creates a standardized baseline for comparison among different study sites, includes PAHs that are not considered to be toxic, and excludes PAHs that appear to be more toxic than any currently on the list (Anderson and Achten, 2015).

Table 2.1

16 EPA Priority Pollutants Ring Structure Compound Abbreviation Acenaphthene AC Anthracene AN 2- ring Naphthalene NA Acenaphthylene ACE Fluorene FL 3-ring Phenanthrene PHE Benzo[a]anthracene B[a]A Chrysene CHR Fluoranthene FA Benzo[b]fluoranthene B[b]F 4-ring Pyrene PY Dibenzo[a,h]anthracene D[ah]A Benzo[a]pyrene B[a]P 5-ring Benzo[k]fluoranthene B[k]F Benzo[ghi]perylene B[ghi]P 6-ring Indeno[1,2,3-cd]pyrene IP Table 2.1 16 EPA PAH Priority Pollutants and their associated abbreviations.

Persistence of PAHs depends on environmental conditions as well as their structure. In general, PAHs will biodegrade best under aerobic conditions, but also have been demonstrated to degrade under anaerobic conditions (Haritash and Kaushik, 2009). However, PAHs will bind to soils with time, making them less bioavailable to microbes and less susceptible to weathering and degradation, leading to their environmental persistence (McDonald and Chapman, 2002).

Therefore, in many field investigations, it is often found that the concentrations of PAHs in soil will decrease rapidly initially, and then plateau in a “hockey stick” shaped curve once the compounds age in a two-phase decomposition. Generally, PAHs that have 2-3 benzene rings or less biodegrade more rapidly than those that are more complicated (National Research Council,

1997). PAHs with 4 or more rings are less available to microbial degradation, volatization and 10 bind to the soil more easily (Neff et al., 2005). Increasing soil temperature has been shown accelerate the loss of PAHs with 3 rings or less from soil (Coover and Sims, 1987). Microbially- aided degradation of PAHs has been documented under a variety of conditions, including psychrophilic, mesophilic (Ma et al., 2006), and thermophilic conditions (Coover and Sims,

1987).

Alkaline Environments

In nature, alkaline environments are relatively understudied and not very well understood. Naturally, alkaline environments are relatively scarce, occurring in only a few soda lakes or hot springs (Horikoshi, 1999). Soda lakes are formed when there is a steady supply of water dissolving calcium and magnesium from sedimentary rocks, coupled with a rapid evaporation rate (Duckworth et al., 1996). These lakes are also often saline as well, providing an additional stressor for microbial life. Soda lake pH values range in between 10-12 (Duckworth et al., 1996).

Alkaline environments are often anthropogenically created, resulting from contamination.

They can form from various sources, including concrete and construction waste (Pederson, et al.,

2004) fly ash (Carlson and Adriano, 1993), and steel manufacture (Roadcap et al., 2005). These sites are not as well studied as their acidic counterparts but are now being researched not only for remediation, but to examine their possibilities as extraterrestrial analogues and their potential for biotechnical applications (Horikoshi, 1999). 11

Anthropogenically created alkaline environments often contain other types of contamination due to their industrial origin. Heavy metals are common (Waska, 2014), and their solubility decreases when pH increases (Fijalkowski et al., 2012). Conversely, the solubility of organic contamination increases with an increasing pH (Yang et al., 2001).

Field Methods of Investigation

Due to the toxicity of hydrocarbons, it is critical to identify the source of the original contamination. Methodologies are often shared with and adapted from oil spill forensics, such as those used after the Exxon Valdez environmental disaster (Page et al., 2002). Hydrocarbon contamination is common in environmental investigations, and thus a variety of approaches are available to characterize both the extent as well as the source of the contaminant.

Establishing a forensic fingerprint is a common way to identify contamination (Boehm,

2006). Hydrocarbons naturally weather into characteristic PAHs, and can be correlated to their parent compound (Neff et. al., 2005).

The 16 EPA priority pollutants (table 2.1) are a common suite of analyates to use when quantifying organic pollution. These 16 analyates contain two isomer pairs, each with a compound that is more stable under high temperatures. By comparing the isomer ratios in a plot, it may be determined if the compounds are petrogenic or pyrogenic in origin. Various statistical tests can also be used to correlate suspected parent materials with a contaminant. A visual comparison of sample profiles on bar charts may be used to qualitatively examine different samples with possible parent products (Boehm, 2006). 12

It is possible to increase the resolution to better determine the origin of contaminants by increasing the number of target analyates sought after. More target analyates have been used in previous investigations to attribute contamination to specific sources, especially through comparison with PCA. With enough analyates, it is possible to distinguish different hydrocarbon sources (Boehm, 2006).

Biological communities can also be used to create fingerprints of sediments and groundwater (Garland and Mills, 1991; Paissé et al., 2008). It is not unusual to find microorganisms that are able to utilize PAHs as their energy source, and many are studied for their value in biodegradation (Haritash and Kaushik, 2009). Microbes are able to break down less alkylated structures with fewer rings more easily than higher molecular weight compounds

(Haritash and Kaushik, 2009).

Community profiles can be used to compare different sampling sites. BIOLOGs provide information on carbon utilization throughout a community (Garland and Mills, 1991). 16S rDNA profiles have also been used to create microbiological fingerprints and compare different field sites. Communities of microbes as a whole will vary based on environmental conditions as well as contamination, and changes between communities can be used to better gage the magnitude of differences between sites (Paissé et al., 2008).

CHAPTER 3

MATERIALS AND METHODS

Sampling Periods

On August 18, 2014, (summer sampling period) an initial transect (figure 2.2) was sampled at Big Marsh and Acme Coke (figure 2.1). Creosote from Acme Coke was obtained for a positive control as well as materials from the nearby railroad tracks. Samples were also obtained from the Slag Dunes to serve as a control, since it is not linked hydrogeologically to

Acme Coke.

On December 16th, 2014, (winter sampling period) the transect was sampled again to augment data gaps found in the summer sampling period. Additionally, control sites at the Slag

Dunes and Wolf Lake were sampled.

Field Sampling Methods

Groundwater, surface water, and sediment samples were collected from the field sites for geochemical and biological tests (table 3.1). Groundwater was obtained from pre-existing wells, where available. The wells were purged (3 well volumes) before sampling with a 0.5 liter plastic bailer. 14

Table 3.1 Collected Field Samples Summary Site PAH Analysis 16S rDNA Biolog Ecoplates Aqueous Matrix Water sediment sequencing Water Sediment Geochemistry Summer Big Marsh Well Groundwater X X X X Big Marsh Spring Surface water X X X X Big Marsh Spring Sediment X Well #3 Slag Dunes Groundwater X X X X RR Ditch Surface water X X X X RR Ditch Sediment X Creosote Marsh Surface water X X X X Creosote Marsh Sediment X Creosote Sediment X

Winter Big Marsh Well Groundwater X X X Big Marsh Well Sediment X X Big Marsh Spring Surface Water X X X RR Ditch Surface Water X X X RR Ditch Sediment X X Creosote Marsh Surface Water X X X Creosote Marsh Sediment X X Well #2 Slag Dunes Groundwater X X X Well #2 Slag Dunes Sediment X X Well #3 Slag Dunes Groundwater X X X Well #3 Slag Dunes Sediment X SD Surface Water Surface Water X X X SD Surface Water Sediment X X Wolf Lake Surface Water X X X Wolf Lake Sediment X X

Table 3.1: Data collected from each sampling site.

Organic Chemistry: PAH Analysis

PAHs samples were collected from water and sediment. Water samples were collected in

1 liter amber glass bottles with Teflon lids. ≥40 grams of sediment were collected in glass jars with Teflon lids provided by First Environmental Laboratories of Naperville, IL according to

EPA protocol SW-846. Samples were immediately placed in a cooler with ice packs for transport to NIU, where they were refrigerated until they were transported to First Environmental

Laboratories on ice.

Samples were processed for the 16 EPA PAH Priority Pollutants by First Environmental.

Water samples used EPA Method 3510C, Separatory Funnel Extraction for Semi-Volatiles (First

Environmental SOP #601). Sediment samples were processed with EPA Method 3540C: Soxhlet

Extraction for Semi-Volatiles (First Environmental SOP #603). After extraction, samples were analyzed using EPA method 8270C: Low Level PNA Compounds by Gas Chromatography/Mass

Spectrometry (GC/MS) (First Environmental method SOP #201).

Aqueous Geochemistry

In the field, temperature, pH, electrical conductivity (EC), oxidation-reduction potential

(ORP), and dissolved oxygen (DO) (DO for winter only, due to a probe malfunction) were measured using a Hach multiprobe HQ 40D. Samples were collected and tested in clean half-liter plastic beakers. Each probe was calibrated the night prior to fieldwork, according to the 16 manufacturer’s standard operating procedures. The pH probe was calibrated with a custom 4 point curve at 4.00, 7.00, 10.00, and 12.43 to reflect the high pH seen in the field. Standard analytical procedure was followed, and prior to testing each sample each probe was rinsed and dried three times with DI-water.

Biological Filtration and 16S rDNA Sequencing

Prior to use, 0.2 μm sterile peristaltic pump filters (Fischer, Waltham, IL) were autoclaved in their housing units and wrapped in aluminum foil. Water was pumped from sites along the transect during the summer sampling period through the sterile filters with a Branant

Company Portable Sampler (Lake Barrington, IL). For each site, 1 liter (or until the filter clogged) of water was filtered. Due to time and safety constraints, water samples from Slag Dune

Well #3 were transported back to the lab in 0.5 liter black Whirlpaks (NASCO, Fort Atkinson,

WI) in a cooler. They were stored in a refrigerator until the next day, when they also could be passed through a filter. Filters were stored at -70°C. The day after the field work, they were removed from filters houses with sterile tweezers and placed in sterile petri dishes (Fischer,

Waltham, IL), which were returned to the -70°C freezer.

After filtration, each filter was weighed with an analytical balance in new, sterile petri dishes in a laminar fume hood. Filters were cut into quarters with scissors and tweezers sterilized with 70% EtOH, and each quarter was weighed individually. The first filter quarter was placed in 1.5 ml Eppendorf tubes (Eppendorf North America, Hauppauge, NY) for storage in a

-70°C freezer. The other three quarters of each filter were placed into individual PowerBead 17

Tubes, following step 1 of the MoBio PowerSoil DNA Isolation Kit (MO BIO Laboratories, Inc,

Carlsbad, CA). DNA was extracted according to the manufacturer’s protocol, with the exception of steps 1-7, which were replaced by placing the PowerBead tubes in a MP Fast Prep24

Homogenizer (MP Biomedicals) at speed of 6.5 for 60 seconds. Extracted DNA samples were stored at 4°C, and PCR amplification of 16S rDNA was performed using materials and methods developed for the Earth Microbiome Project (Caporaso et. al, 2012).

A negative control blank without DNA added was also processed to verify that quality control standards were met. The PCR products were then run on a 1% Ethidium Bromide agarose gel with a ladder to check amplification success. To accurately quantify DNA concentration, 1μL of each PCR product was read on a Nanodrop 1000 Reader (ThermoFisher Scientific, Waltham,

MA). From each triplicate, the sample that had the highest ng/μL DNA concentration, as well as the 260/280 ratio closest to 2, was sent to the University of Illinois at Chicago for Illumina

MiSeq 16S rDNA sequencing with 150 bp paired reads.

BIOLOG Ecoplates

Samples for Biolog Ecoplate analysis were collected for both the summer and winter sampling seasons. Water samples were collected in 15ml Falcon tubes (Corning, Tewksbury,

MA). Sediment samples were collected in 0.5 liter black Whirlpak bags using sterile, autoclaved tongue depressors. Samples were stored in a refrigerator until they were plated.

Approximately 1.00g of soil was weighed with sterile, autoclaved tongue depressors into

15ml Falcon tubes. Each sample tube then had 12ml of autoclaved Phosphate Buffer Solution 18

(PBS) added, and vortexed for 10 minutes. After vortexing, the samples were centrifuged in a

Marathon 21000R (ThermoFisher Scientific, Waltham, MA) at 25000R rpm for 20 minutes at

25°C. After centrifugation, 10ml of the supernatant is transferred into new 15ml Falcon tubes.

The sediment extracts were refrigerated at 5°C overnight until being plated in the morning.

The BIOLOGs were plated next to an open flame to help maintain sterility and prevent contamination. Prior to plating, each sample was vortexed for 30 seconds, and the benchtop, gloves, and pipette handle are wiped down with 70% EtOH. Each well is inoculated with 100μl of sample.

Using the Epoch Biotek Spectrophotometer (BioTek US, Winooski, VT), carbon utilization and microbial activity can be quantitatively documented. Plates were read at day 5 at a wavelength at 690nm. Due to technical difficulties, the winter BIOLOG plates were not run until five days after sample collection.

CHAPTER 4

DATA ANALYSIS AND DISCUSSION

Results were obtained through biological as well as organic and inorganic investigative methods. Table 3.1 summarizes the methods used for each sample.

Inorganic Chemistry

Geochemical measurements were taken in the field for both ground and surface water during the summer sampling period (table 4.1). Both the RR Tracks as well as Creosote Marsh have circumneutral pHs, whereas Big Marsh and the Slag Dunes have hyperalkaline pHs.

Groundwater samples as well as alkaline surface water samples have negative oxygen-reduction potential (ORP). The Creosote Marsh and the RR Ditch have the lowest electrical conductivity, whereas groundwaters from Big Marsh show an electrical conductivity of 9900 µS/cm.

Table 4.1 Summer Water Sample Inorganic Characterization (8/18/2014) Big Big Marsh Surface Well #3 Creosote Site Units Well Water slag dunes RR Ditch Marsh pH 12.75 11.83 12.32 7.86 8.49 Temp [C°] 17.13 27.7 19.48 26.3 28.7 ORP [mV] -172.8 -65.2 -261.9 102.8 144.2 EC [µS/cm] 9900 2300 6160 1006 768 Table 4.1 Summer groundwater and surface water samples. 20

The winter water geochemistries (table 4.2) are similar to the summer, with some exceptions. As observed in Waska (2014), pH tends to increase in the winter months. The circumneutral sites Big Marsh, Wolf Lake, and the RR Ditch all have a pH that exceed 9. ORPs are similar to the summer sampling period, with groundwaters having reducing conditions.

Surface waters have dissolved oxygen levels around 10 mg/L. The groundwater samples have dissolved oxygen levels much lower, at 4 mg/L. As noted in Waska (2014), with less water in the geologic system, the electrical conductivity and pH increases in the winter months, with Big

Marsh Well, Creosote Marsh, and the Slag Dunes each having an EC well above average seawater (California State Water Control Board, 2016). As with the winter months, all ORPS of groundwater show reduction potential with negative values.

Depending on the ORP values, different microbial processes may occur. Positive ORP values can lead to the reduction of iron, manganese, and nitrate (Bier, 2009), processes that are all likely occurring at the RR Ditch and Creosote Marsh in the summer and winter. The Slag

Dune surface water, however, has a negative ORP, distinguishing it from the other sites. An ORP in the ranges of 100 to -200 leads to the reduction of sulfate, and ORPs ranging between -150 to

-280 leads to the creation of methane (Bier, 2009). Each of the groundwaters as well as the surface water at the Slag Dunes and Big Marsh show negative ORPs. The potential to create sulfate is present at Big Marsh Well (summer and winter), Big Marsh Spring (summer and winter), the Slag Dune Surface Water, and the Slag Dune Well #3 (winter). The potential to create methane is present in multiple samples, including Big Marsh Well and Well #3 Slag

Dunes in both the summer and winter sampling periods. 21

Table 4.2 Winter Water Sample Inorganic Characterization (12/16/2014) Slag Big Big Well #2 Well Dune Marsh Marsh RR Creosote Slag #3 Slag Surface Wolf Site Units Well Spring Ditch Marsh Dunes Dunes Water Lake pH 13.43 12.73 9.26 9.55 13.14 13.11 12.65 9.84 Temp [C°] 8.825 4.525 2.475 2.25 7.7 8.8 5.2 3.8 ORP [mV] -165.1 -58.4 56.2 56.8 -101.4 -288.5 -131.3 91.7 EC [µS] 10,200 3800 773 5650 6680 6760 1522 1435 LDO [mg/L] 4.19 9.2 3.4 11.43 3.31 3.91 10.76 12.65 Table 4.2: Winter ground and surface water samples.

Organic Geochemistry

During both the summer and the winter sampling periods, sediment and water samples were taken from the transect, Wolf Lake, as well as the Slag Dunes. Samples were collected for

16 EPA Priority Pollutants analysis (tables 4.3 and table 4.4). At multiple sites, many or all of the analyates tested for were under the detection limit, especially in groundwater and surface water. This may be attributed to the low solubility of PAHs (Joa et al., 2009), preventing their dissolution into the water column once adsorbed onto sediment and decreasing bioavailibility

(McDonald and Chapman, 2002). This is common for legacy contamination sites and can inhibit biodegredation (Davenport and Spacey, 2001).

Table 4.3 Summer 16 EPA PAH Priority Pollutant Data (8/18/2014) detection Big Big Slag Big RR Creosot RR Creosote unit (SED; limit Creosote Marsh Marsh Dune Marsh Track e Marsh Ditch Marsh WATER) (SED; SED Well Surface Well SED SED SED Water Water WATER) Water Water #3 AC [µg/kg; µg/L] 50; 0.25 UD UD 1.55E+06 2.09E+03 UD UD UD UD 1.56E+00 ACN [µg/kg; µg/L] 50; 0.25 UD UD 4.49E+05 1.63E+03 UD UD UD UD 5.27E+00 AN [µg/kg; µg/L] 50; 0.25 9.20E+01 8.30E+01 3.30E+06 5.76E+03 3.40E-01 UD UD UD 1.70E+00 B[a]A [µg/kg; µg/L] 8.7; 0.25 3.40E+02 2.99E+02 4.17E+06 1.43E+04 UD UD UD UD UD B[a]P [µg/kg; µg/L] 15; 0.25 3.64E+02 4.76E+02 5.37E+06 2.41E+04 UD UD UD 2.70E-01 UD B[b]F [µg/kg; µg/L] 11; 0.25 3.40E+02 4.97E+02 4.66E+06 1.52E+04 UD UD UD UD UD B[k]F [µg/kg; µg/L] 11; 0.25 2.93E+02 2.70E+02 3.52E+06 1.91E+04 UD UD UD UD UD B[ghi]P [µg/kg; µg/L] 50; 0.25 1.68E+02 3.06E+02 1.38E+06 8.95E+03 UD UD UD UD UD CHR [µg/kg; µg/L] 50; 0.25 2.52E+02 3.11E+02 3.43E+06 1.28E+04 UD UD UD UD UD D[ah]A [µg/kg; µg/L] 20; 0.25 2.70E+01 UD 1.15E+06 5.70E+03 UD UD UD UD UD FA [µg/kg; µg/L] 50; 0.25 7.18E+02 5.24E+02 5.72E+06 1.66E+04 UD UD UD 3.00E-01 1.09E+00 FL [µg/kg; µg/L] 50; 0.25 UD 5.70E+01 2.85E+06 2.58E+03 UD UD UD UD 4.08E+00 IP [µg/kg; µg/L] 29; 0.25 2.01E+02 2.59E+02 4.04E+06 2.16E+04 UD UD UD UD UD NA [µg/kg; µg/L] 25; 0.25 3.70E+02 2.20E+02 5.77E+06 1.27E+04 1.54E+01 1.30E+00 UD UD 7.30E+01 PHE [µg/kg; µg/L] 50; 0.25 3.81E+02 3.16E+02 5.96E+06 1.31E+04 6.40E-01 3.50E-01 UD UD 1.39E+01 PY [µg/kg; µg/L] 50; 0.25 5.85E+02 4.74E+02 4.80E+06 1.40E+04 2.90E-01 UD UD 2.50E-01 5.60E-01 Total [µg/kg; µg/L] 50; 0.25 4.13E+03 4.09E+03 5.81E+07 1.90E+05 1.67E+01 1.65E+00 0.00E+00 8.20E-01 1.01E+02 PAHs Notes: UD = Under the Detection Limit, SED=Sediment Table 4.3: Summer groundwater, surface water, and sediment found at each sampling site.

Table 4.4 Winter 16 EPA PAH Priority Pollutant Data (12/16/2014) Slag Slag Slag Slag Slag Slag unit of detection Big Dune Dune Dune Big Dune Big Dune dune measureme limit RR Marsh Surface Well Well Creosote Marsh Wolf RR Big Wolf Well Marsh Creosote Well Surface nt (SED; (SED; Ditch Well Water #2 #3 Marsh Spring Lake Ditch Marsh Lake #2 Well Marsh #3 water WATER) WATER) WAT WAT WAT WAT WAT WAT WAT WAT SED SED SED SED SED SED SED SED [µg/kg; AC µg/L] 50; 0.25 UD UD UD UD UD UD UD UD UD UD UD UD UD 8580 UD UD

[µg/kg; ACN µg/L] 50; 0.25 UD UD UD UD UD UD UD UD UD UD UD UD UD 5530 UD UD [µg/kg; AN µg/L] 50; 0.25 UD UD UD UD UD UD UD UD 484 90 UD UD 115 23100 UD UD

[µg/kg; B[a]A µg/L] 8.7; 0.25 0.34 UD UD UD UD 1.11 UD UD 2070 377 34.2 92.5 451 80500 193 86.7 [µg/kg; B[a]P µg/L] 15; 0.25 0.4 UD UD UD UD 1.8 UD UD 1990 345 44 129 471 137000 213 69

[µg/kg; B[b]F µg/L] 11; 0.25 0.51 UD UD UD UD 1.73 UD UD 1640 247 45 170 317 88300 226 69 [µg/kg; B[k]F µg/L] 11; 0.25 0.26 UD UD UD UD 0.83 UD UD 1970 306 42 103 443 122000 193 64

[µg/kg; B[ghi]P µg/L] 50; 0.25 UD UD UD UD UD 1.4 UD UD 1660 224 UD 148 424 88300 188 54 [µg/kg; CHR µg/L] 50; 0.25 UD UD UD UD UD UD UD UD 2190 314 UD 127 558 96100 188 81

[µg/kg; D[ah]A µg/L] 20; 0.25 UD UD UD UD UD 0.3 UD UD 309 63 UD 38 127 24400 44 UD [µg/kg; FA µg/L] 50; 0.25 UD UD UD UD UD UD UD UD 4370 612 73 186 946 95400 306 251

[µg/kg; FL µg/L] 50; 0.25 UD UD UD UD 4 UD UD UD 290 UD UD UD 51 9150 UD UD [µg/kg; IP µg/L] 29; 0.25 0.3 UD UD UD UD 1.5 UD UD 1410 228 41 142 340 108000 177 56

[µg/kg; NA µg/L] 25; 0.25 UD 24 UD 19 140 UD UD UD 193 490 28 UD 313 36200 69 61 [µg/kg; PHE µg/L] 50; 0.25 UD UD UD UD 16 UD UD UD 2590 373 61 58 740 56600 139 246

[µg/kg; PY µg/L] 50; 0.25 UD UD UD UD UD UD UD UD 3170 545 96 164 764 74800 255 195

Total [µg/kg; PAHs µg/L] 50; 0.25 1.81 24 0 19 160 8.67 0 0 24336 4214 464.2 1358 6060 1E+06 2191 1232.7

Notes: UD = Under the Detection Limit, SED=Sediment WAT=Water

Table 4.4: Surface water, groundwater, and sediment PAHs.

The Pearson Product-Moment Coefficient was calculated in Microsoft Excel to further compare the samples (Hezong et al., 2015). The Pearson Product-Moment can be used to calculate the R value to test the relationship of one array of data to another. The closer the data is to 1, the higher the correlation. Data between [0.5]-[1] has a strong correlation, [0.3]-[0.5] has a moderate correlation, and [0]-[0.3] is a weak correlation. Raw creosote was compared to the 16

PAH profiles from each sampling site (tables 4.5 and 4.6). In table 4.5, the sediment from the

Winter PAHs show a strong positive correlation the Big Marsh Sediment, as well as the RR 24

Ditch and Creosote Marsh. Big Marsh surface and well waters, Creosote Marsh surface water, as well as the Slag Dune well water all show a moderate positive correlation to the control sample.

Table 4.5 Summer PAH Pearson Coefficients (12/16/2014) Slag Big Big RR Creosote Big RR Creosote Dune Marsh Marsh Ditch Creosote Marsh Marsh Ditch Marsh Well Surface Well Surface SED Surface SED SED SED Water Water Water Water Water #2 Pearson 0.41 0.82 0.34 0.74 0.00 1.00 0.71 0.47 0.35 Coefficient Table 4.5: Correlations between samples and a Creosote Sediment Control.

Winter PAHs profiles (table 4.6) were compared to sediment from Creosote Marsh. The

Sediment from Big Marsh, Big Marsh Well, sediment and water from the RR Ditch, Creosote

Marsh water, Slag Dune Well #2 Sediment, and the Wolf Lake Sediment all indicate a strong positive correlation with the Creosote Sediment. The Slag Dune Surface water sediment showed a moderately positive trend, and Big Marsh Well water as well as Slag Dune Wells #2 and #3

Water did not show any correlation with the Creosote Sediment. All sediment samples (with the exception of the Slag Dune Surface Water) in the Pearson Correlation show a strong R correlation to the creosote control. While this suggests that contamination at Big Marsh may have come from the Acme Coke, the correlation at the control sites is similar to that of Big

Marsh. This implies that the PAH contamination is more widespread than groundwater could be expected to transport.

Table 4.6 Winter PAH Pearson Coefficients (8/18/2014) Slag Slag Slag Slag Slag Slag Big Big Big Big RR RR Creosote Creosote Dune Dune Dune Dune dune Dune Wolf Wolf Marsh Marsh Marsh Marsh Ditch Ditch Marsh Marsh Well Well Well Well Surface Surface Lake Lake Spring Well Well SED SED water SED water #2 #2 #3 #3 water Water SED water water SED water SED water SED water SED water Pearson 0.6 0 0.65 -0.18 0.69 0.62 1 0.67 0.84 -0.18 0.87 -0.2 0.42 0 0.52 0 Coefficient Table 4.6: PAH correlations between winter water and sediment samples to Creosote Marsh samples.

PAH data can be further examined through the use of PAH diagnostic double ratio plots.

With larger suites of PAH analyates, it is possible to differentiate between separate sources of contamination through statistical techniques such as PCA (Neff et al., 2005; Brenner et al.,

2002), creating carbon fingerprint profiles (Alimi et al., 2003), as well as positive matrix factorization (PMF) (Sofowote et al., 2008). However, with only 16 EPA Priority Pollutants, the highest resolution that can be obtained is determining whether compounds are petrogenic or pyrogenic in origin by plotting them on a graph (figure 4.1) with isomer ratios (Maher and

Aislabe, 1990; Neff et al., 2005)

Phenanthrene and anthracene are isomers, as are fluoranthene and pyrene. In each isomer pair, there is a compound that is not stable under high temperatures (Maher and Aislabe, 1990;

Neff et al., 2005). Thus, by comparing the ratio phenanthrene to anthracene to (PH/AN) and fluoranthene to pyrene (FL/PY), it can be determined if a compound is petrogenic (formed through normal geologic processes) or pyrogenic (formed through combustion) in origin (Maher and Aislabe, 1990; Neff et al., 2005).

In figure 4.1, all compounds that are on the sampling transect are clustering near one another, seeming to indicate that they share parent product that has a similar origin. This comparison was limited by a number of samples where one or all of phenanthrene, anthracene, fluoranthene, or pyrene were under the detection limit, especially in groundwater and surface water samples. Since phenanthrene and anthracene are both lower molecular weight molecules, they are quicker to weather while other PAHs remain. 26

Figure 4.1 Isomer ratio Analysis of each site, compared to known PAH profiles (Neff et al., 2005) In figure 4.1, it can be observed that all compounds are likely pyrogenic in origin based on their location on the lower left hand side of the graph. However, the Slag Dunes are represented away from the rest of the group, indicating that their parent compound may be at least partially different than other compounds on the transect.

Gray lines on the chart represent known PAH contamination from literature (Neff et al.,

2005). All the samples from the transect cluster around diagnostic ratios known from coke oven emissions. Coke oven emissions come from the air, potentially indicating that the contamination may have been airborne and is not necessarily being transported from groundwater. This is consistent with the Pearson correlations, which indicate that the PAH contamination is more widespread than expected. 27

Using known slag waste bulk density (FHA, 1997) and estimated total carbon (Wiener,

2014), the retardation factors (table 4.7) for each Priority Pollutant were calculated using equations 4.1-3 (Wiener, 2014).

푙표푔퐾표푐 = 0.983푙표푔퐾표푤 + 0.00028 [4.1]

퐾푑 = 퐾표푐푓표푐 [4.2]

휌퐾 푅 = 1 + 푑 [4.3], ℎ

Where 퐾표푐 = organic carbon-water partitioning coefficient,

퐾표푤= octanol-water partitioning coefficient,

퐾푑= distribution coefficient,

푓표푐 = Mass Fraction of Organic Carbon =4.0% (Wiener, 2012)

ℎ = effective soil porosity = 0.35 (Wiener, 2012)

휌 = soil bulk density= 1.24g/푐푚3 (FHA, 1997)

푅= Retardation factor

Retardation factors of less than 3 are considered very mobile, 3-9 are mobile, 30-100 are less mobile, and retardation factors greater than 100 are considered immobile (Wiener, 2012).

Chrysene is the only very mobile priority pollutant, and naphthalene, acenaphthylene, and fluorine are mobile. These contaminants are the most likely to be transported through the groundwater. Overall, most of the Priority Pollutants are not mobile and PAH contamination is not likely to be spreading to a great extent through the groundwater.

Table 4.7

16 EPA Priority Pollutant Mobility Ring

Structure Compound R

2- ring Acenaphthene 3.92 3.85 2.86 11.12 Anthracene 4.54 4.46 11.62 42.16 Naphthalene 3.37 3.31 0.82 3.91 3-ring Acenaphthylene 4 3.93 3.42 13.13 Fluorene 4.18 4.11 5.14 19.22 Phenanthrene 4.57 4.49 12.44 45.06 4-ring Benzo[a]anthracene 5.91 5.81 258.15 915.58

Chrysene 1.65 1.62 0.02 1.06 Fluoranthene 5.22 5.13 54.15 192.85 Benzo[b]fluoranthene 5.8 5.7 201.25 714.01 Pyrene 5.18 5.09 49.46 176.24 5-ring Dibenzo[a,h]anthracene 6.75 6.64 1728.18 6123.71 Benzo[a]pyrene 6.04 5.94 346.47 1228.48 Benzo[k]fluoranthene 6 5.9 316.48 1122.23 6-ring Benzo[ghi]perylene 6.5 6.39 981.39 3477.91 Indeno[1,2,3-cd]pyrene 6.58 6.47 1176.20 4168.10 Table 4.7: Mobility of the EPA Priority Pollutants.

Biology

Biological for samples were taken during both the winter and summer sampling periods.

During the summer sampling period, only ground and surface water samples were taken for

BIOLOG culturing. Additionally, samples were taken for 16s rDNA sequencing and bioinformatics analysis. During the winter sampling period, BIOLOGs were inoculated with microbes from sediment and water samples. Carbon utilization profiles for each site were investigated through the use of BIOLOG Ecoplates, which have 21 possible carbon sources for 29 microbes (table 4.8). This can be used to create community carbon utilization profiles for each sampled site (Garland and Mills, 1991).

Table 4.8 Structure of BIOLOG Carbon Substrates POLYMERS AND OTHER CHAIN COMPLICATED STRUCTURES Malic Acid Glycogen Pyruvic acid alpha-CYCLODEXTRIN Putriscene Tween 80 L-arginine Tween 40 L-asparagine SINGLE RING Glycyl-L-glutamic acid Phenethylamine L-serine L-phenylalanine L-threonine N-Acetyl-beta-D-glucosamine i-Erythritol beta-Methyl-D-glucoside D-mannitol D-Xylose alpha-keto-butyric acid 2 Hydroxybenzoic Acid Itaconic Acid 4 Hydroxybenzoic Acid DL-Alpha-Glycerol Phosphate D-Gluconic acid, gamma-lactone D-Galacturonic Acid Glucose-1-phosphate D-Glucosaminic Acid DOUBLE RING Gamma Hydroxybutyric Acid D-Cellobiose alpha-D-lactose Table 4.8: Carbon substrates in Biolog Ecoplates, grouped by structure.

In figure 4.2, the sum of all carbon sources for each sampling location is presented.

Plates that had no growth were omitted. With the exception of Slag Dune Well #3, none of the groundwater samples cultured any microbial growth. This may be due to the changes in environmental conditions from the groundwater to the surface, especially the sudden introduction of oxygen into an environment where microbes would otherwise be living in near anoxic conditions. All surface and groundwaters from Big Marsh showed no significant growth in the

BIOLOG plates, but there was some growth in the Big Marsh winter sediment sample. Slag

Dune surface water and groundwater in the winter time both showed minimal growth compared 30 to other samples. The RR Ditch had the most growth in every sample compared to other sample sites.

A regression analysis in Microsoft Excel reveals a direct correlation between microbial growth and pH. Overall, pH correlates directly to microbial growth with an 푟2 = 0.50, where high pH inhibits microbial growth (figure 4.3).

BIOLOG Total Microbial Growth 60 50 40 30 20

Microbial Growth 10 0

Site

Figure 4.2: Sum of Microbial Growth on carbon source for each site. GW= Groundwater, SW=Surface Water, SED=Sediment, WIN= Winter Samples, SUM=Summer samples.

pH Related to Microbial Growth 60 R² = 0.5033 50

10 Total Microbial Growth 0 0 2 4 6 8 10 12 14 16 -10 pH

Figure 4.3: Regression analysis of pH and microbial growth. Slag Dune Well #3 was omitted due to apparently anomalous data. These profiles were compared using principal component analysis (PCA) (R, 2013).

Average microbial growth for each carbon source was normalized by the average of the control wells. Carbon sources with growth less than 0.5 were considered non-growth, and were replaced by zero in PCA analysis (Raimondi, 2015; Greer, 2014). 32

BIOLOG PCA Data

3 Big Marsh 2 Creosote Marsh 1 RR Ditch 0 Slag Dunes

-1010.8% -5 0 5 10 -1 Wolf Lake Sediment -2 Groundwater -3 Surface Water -4 56.5%

Figure 4.4: PCA of Biolog Growth. Circles represent surface water, diamonds represent groundwater, and squares are sediment samples. Open shapes are summer samples, and closed are winter samples.

Differences when comparing all summer and winter samples (figure 4.4) appear to be dominated by pH. The high pH of Big Marsh and the Slag Dunes (red and blue, respectively) are grouped to the left of the graph, and circumneutral pH sites of Creosote Marsh, RR Ditch, and

Wolf Lake (green, yellow, and purple, respectively) are grouped to the right. Surface water

(circles) samples are clustered in the lower right side of the graph, and sediment samples

(squares) are in the upper center, seeming to indicate that sample matrix explains other differences in microbial growth.

BIOLOGS were also analyzed by carbon structure (figure 4.5, table 4.8). Simpler, lighter organic compounds break down more easily than more complicated structures (Amon and

Benner, 1996). Additionally, steric effects cause an inhibition of microbes from accessing their reaction sites (Maier, 2009). Single and double ring structures are the most similar to the 16 EPA

Priority Pollutants, and are therefore the focus of this investigation. 33

BIOLOG Carbon Structure Utilization 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 0%

chain polymer double ring single ring

Figure 4.5: Biolog carbon Utilization by Carbon Source Structure.

The preference of microbes for single and double ring structures does not appear to be correlated to the number of PAHs present in their environment. Even though it would be expected that Creosote Marsh would have the highest concentration of total PAHs (table 4.9), its microbial community has one of the lowest percent utilization of single/double ring structures.

All samples show the ability to metabolize single structures; however, Big Marsh Well sediment and Slag Dune Well #3 do not have the ability to break down double ring structures.

Table 4.9 Total PAHS of Selected Sites Big unit of Marsh RR RR Creosote Slag measurement GW RR Ditch Ditch Ditch Marsh Creosote Creosote Dune Slag Dune Wolf (SED; (SED) (SED) SW SW SED Marsh Marsh SW SW Well #3 Lake SED Wolf Lake WATER) WIN WIN SUM WIN WIN SW SUM WIN WIN GW WIN WIN SW WIN Total [µg/kg; PAHs µg/L] 6.06E+03 4.21E+03 0 1.81 1.05E+06 8.20E-01 8.67 0 1.60E+02 4.64E+02 0

Table 4.9 Total PAHs of sites that had positive BIOLOG growth. 34

To create a more complete community profile for each site, 16S rDNA was extracted from each of the summer water samples. The BIOLOG data illustrates what is live and viable at the time of sampling, but represents only a small number of environmental microbes that can be cultured in lab (Pham and Kim, 2012). A 16S rDNA community profile (figure 4.6) gives insight to microbes that are not represented in BIOLOGs.

Summer 16S rDNA Microbial Profiles by Class

Figure 4.6: 16S rDNA profiles for each summer sampling site. Due to their prevalence of oil spills, hydrocarbon degrading bacteria are often studied in in marine environments rather than the freshwater environments (Jin et al., 2014). However, ocean environments contaminated with hydrocarbons have yielded similar profiles to the ones in figure 4.6, with Proteobacteria, Bacteriodes, and Actinobacteria comprising the majority of the communities (Paissé et al., 2008). 35

The RR Ditch and the Creosote Marsh surface waters both have the most similar 16s rDNA profiles. Both are comprised of a large percentage of Actinobacteria, which as a class are known to occur widely in soil and aquatic environments. Actinobacteria includes characteristic soil bacteria such as Streptomyces (Goodfellow, 1997). This is also shared in the Big Marsh

Well, and to small extent the Slag Dunes Well #3.

Classes of the phylum Bacteroides were best represented by Flavobacteria, Cytophagia, and Spinghobacteria, are seen in just the RR Ditch and Creosote Marsh sample and are suspected hydrocarbon degraders (Mueller et al., 1989; Haritash and Kaushik, 2006). Similarly to profiles in Figure 4.5, Bacteroides have been seen to make up a significant portion of communities that are contaminated by hydrocarbons (Patel et al., 2016). Large populations of Flavobacteria and

Cytophagia, like those seen in the RR Ditch and Creosote Marsh, are found consistently at other terrestrial sites contaminated by hydrocarbons (Popp et al., 2016), as well as seawater (Brakstad et al., 2004).

The Alphaproteobacteria have proven to be vital in hydrocarbon degradation as well, with the genus Azospirillum proving to be critical to creosote degradation in laboratory studies

(Vinas et al., 2005), and being a significant community portion of contaminated microcosm communities (Martin et al., 2012). In microcosm studies of hydrocarbon contaminated soils,

Betaproteobacteria has been seen to increase in the community, becoming the majority class in soils that are inoculated with anthracene (Martin et al., 2012) and important creosote-degraders

(Vinas et al., 2005). Betaproteobacteria, as a class, comprises of a wide range of soil decomposers, like Actinobacteria. Betaproteobacteria have been known to dominate soil samples that are contaminated with hydrocarbons (Patel et al., 2016). Community shifts are also seen in 36 environmental samples, where Betaproteobacteria and Gammaproteobacteria were only seen in contaminated soils not when compared to uncontaminated soils from the same area (Labbe et al.,

2007).

The Big Marsh profile is majorly composed of both the Gammaproteobacteria, and

Betaproteobacteria. The bacteria of the Slag Dunes and the Big Marsh Well has a large variety of

Firmicutes, which are commonly associated with extremeophiles (Schieifer, 2009). Big Marsh also has a large percentage of Cyanobacteria, photosynthesizers that are part of a healthy food chain. Finding the large variety of Firmicutes is consistent with the low dissolved oxygen found at this site (Schieifer, 2009). Firmicutes have been the majority class in communities in sediment contaminated with PAHs from E-waste (Zhang et al., 2010) as well as from PAH-contaminated agricultural soils (Ma et al., 2010) Firmicutes were seen to make up a large portion of communities, along with Bacteriodes, and Actinobacteria Proteobacteria in hyperalkaline locations (Lopez-Garcia et al., 2005). Similarly, in another study, Firmicutes,

Alphaproteobacteria, and Bacteroides dominated DNA profiles from alkaline lakes (Mesbah et al., Wiegel, 2007).

The 16S rDNA profiles were examined with principal component analysis through the use of R (R, 2013) and Excel (figure 4.7), confirming observations in figure 4.6. On the class level, the high pH Slag Dune Well and Big Marsh Surface water sites are the most closely related. This does not appear to be related to any of the geochemical data collected or PAH concentrations, but instead appears to be related to diversity. 37

16S rDNA Profiles PCA 0.6 Creosote Marsh SW 0.4 Big Marsh GW 0.2 RR Ditch SW 0 -0.6 -0.4 -0.2 0 0.2 0.4 0.6

20.1% -0.2

-0.4 Big Marsh SW -0.6 Slag Dune GW

-0.8 64.6%

Figure 4.7 Principal Component Analysis for Taxonomic Groups by Site

Equations 4.5, 4.5, and 4.6 (Hill et al., 2003) are used to describe diversity in table 4.10.

These indices describe the evennesss of the distribution of individuals in a population better than the richness alone (total number of individuals in a population) (Hill et al., 2003).

푛 푛 퐻′ = − ∑ 푖 푙푛 푖 [4.4] 𝑖=1 푁 푁

∑푠 푛 (푛 −1) 퐷 = 1 = 푖=1 푖 푖 [4.5] 푁(푁−1)

퐻′ 퐸 = [4.6] 퐻 푙푛푆

Where 푛𝑖= number of individuals;

N= total population

S= total number of species in a community (richness)

H’= Shannon Wiener Diversity Index

D= Simpson’s Diversity Index

E= Shannon’s Equibility 38

Overall, Creosote Marsh has the most OTUs of microbes from 16S rDNA profiling, while the RR Ditch has the sparsest number of OTUs. Diversity indices for the class data were taken for each of the sample sites. Populations were nearly evenly diverse, with Creosote Marsh showing the least species diversity and the well at Big Marsh showing the highest diversity.

Species richness was the lowest in Big Marsh, and highest at Creosote Marsh. While Creosote

Marsh has the highest population richness, it has the lowest diversity. This is likely the

Actinobacteria (figure 4.6) dominating Creosote Marsh’s 16S rDNA profile. Big Marsh Well, despite its extreme conditions, has the highest equability and evenness for each of the samples.

The class grouping in figure 4.7 appears to be caused not necessarily by pH or matrix, as the

BIOLOGS would seem to be indicate, but instead by diversity. Both Big Marsh surface water and the Slag Dune groundwater have the lowest species richness.

Table 4.10 Diversity Indices (from Class Data) Big RR Marsh Big Marsh Ditch Creosote Slag Dune Value (SW) Well (GW) (SW) Marsh (SW) Well (GW) Total OTUs 93940 117339 85912 178240 108583 Richness 47 62 65 73 52 H’ 1.47 1.69 1.73 1.51 1.55 퐸퐻 0.38 0.41 0.42 0.35 0.39 D 0.71 0.77 0.73 0.65 0.70

Table 4.10: Population Diversity for each 16S rDNA profile.

16S rDNA was processed by using the open software program PICRUSt, which associates certain sections of DNA with presumed biologic functions, referred to as Kyoto

Encyclopedia of Genes and Genomes (KEGG) pathways (Langille et al., 2013). PICRUSt was used this to find microbes that have known KEGG pathways attributed to PAH degradation. 39

Xenobiotic pathways were focused on for this analysis, and pathways were grouped by like compound groups (ex: Atrazine and DDT degradation were grouped together as “pesticides”)

(figure 4.8).

While PAH degradation was an outright KEGG pathway, other pathways are also associated with PAH degradation as well. Naphthalene degradation, an EPA Priority Pollutant, has its own KEGG pathway. Cytochrome P450 is thought to be the primary enzyme responsible for PAH degradation (Das and Chandran, 2011). BTEX (Pollard et. al, 1994) and styrene

(Rosenfeld and Plumb, 1991) are organic compounds that might be expected to be found at hydrocarbon-contaminated sites, similar to the field sites.

The PICRUSt profiles for each site were very similar, showing communities with the same predictive abilities to metabolize PAHs and compounds associated with hydrocarbon contamination.

PAH-Related KEGG Pathways 7 6 5 4 3 2 1 0

Normalized KEGG Pathways Big Marsh Big Marsh RR Ditch Surface Creosote Marsh Slag Dune Well Surface Water Groundwater Water Surface Water #3 Groundwater Sample

Other Organics PAH Napthalene Styrene Cytochrome P450

Figure 4.8: Xenobiotic KEGG Pathways from PICRUSt for each site from 16S rDNA profiles, normalized by total 16S rDNA OTUs. 40

Figure 4.8 suggests that Creosote Marsh and the RR Ditch have a relatively higher proportion of microbes able to break down PAHs and other compounds associated with hydrocarbon contamination. Data for PAH pathways and other hydrocarbon pathways was normalized against the total population (table 4.11). A one-sample T-test was performed where alpha= 2.776 to determine if any of the samples is significantly different (Ambrose III et. al,

2007). The only sampling site with a significantly different percentage of KEGG pathways was the RR Ditch. While the RR Ditch does have relative high total PAHs present, it does not have the highest of all the samples. However, it does have the lowest pH of the sites tested in both the summer and winter time. As seen in figure 4.2, communities with environments with pHs closest to neutral had the most microbial activity, therefore, it is reasonable that a community with the lowest pH but a high PAH contamination would have the highest percentage of microbes capable of breaking down hydrocarbons.

Table 4.11 PICRUSt PAH KEGG Pathways One-Sample T-test Big Slag Big Marsh RR Creosote Dune Marsh Surface Ditch Marsh Well Well water #3 T-value PAH KEGG Pathways -1.447 -0.474 3.843 -0.230 -1.692 T-value Other PAH KEGG Pathways 0.974 -0.585 3.396 -1.835 -1.951 Table 4.11: T-test comparing samples, where α=2.776.

A regression analysis in Excel examining the relationship PAH degrading pathways and total PAHs in the sediment revealed a positive trend (figure 4.9). Due to the low solubility of the

PAHs, PAH measurements of sediments were used. The total KEGG pathways for each site can be correlated to total PAHs present in the sediment samples of each site. There is a strong trend 41 suggesting that the total PAHs influence the number of hydrocarbon-degrading pathways present in microbial communities, with an 푅2=.78.

Hydrocarbon Degradation KEGG pathways Related to Total PAHs 30000

25000

20000 R² = 0.7794

15000

10000 Total PAHs 5000

0 0 2 4 6 8 10 12 14 16 -5000 Normalized KEGG hydrocarbon-degrading pathways

Figure 4.9: Total PAH sediment samples in relation to normalized PAH-degrading pathways. Due to the magnitude of difference in total PAHS of Creosote marsh to the second highest site, it was not used for the regression analysis.

CHAPTER 5

CONCLUSION

The primary purpose of this investigation was to determine if hydrocarbons are being transported by the groundwater from the former site of Acme Coke downgradient into the Big

Marsh area. Additionally, this research attempted to characterize this unique microbial community at these sites, as well as the relationship of the microbial communities to the PAH contamination.

Due to the small target analyate list for the hydrocarbons, it is very difficult to attribute an exact parent product for the PAH contamination. However, compound profiles are consistent with creosote samples taken from Acme Coke. While it is clear that the contamination is pyrogenic in origin, and it is likely that the contamination came from the air and is not being transported through the ground and surface waters.

Biological analysis indicated the microbial growth is strongly influenced by pH. High pH sites have microbes that do not grow as well in a laboratory setting. The matrix that the bacteria originated from also strongly influenced the microbial community. All communities sampled

(that had significant growth) had the ability to utilize single or double ring structures similar to

PAHs when cultured. This was confirmed by 16S rDNA analysis, where each community had microbes possessing pathways attributed to hydrocarbon degradation. Additionally, the Slag

Dune Well #3 surface water as well as the Big Marsh surface water seem to have microbial communities that are the most influenced by PAHs, based on their 16S rDNA profiles. 43

This investigation suggests that future work in the Calumet area should focus on the origin of PAHs. It is understood that these PAHs are likely not mobile in the groundwater from the Acme Coking Facility. Further investigations should focus on identifying parent compounds more definitively by use of larger suites of analyates or isotopic analysis. Biological investigations should search for groundwater and surface water that is uncontaminated by PAHs to better understand the role of microbes on PAH degradation. However, because of the amount of industrial activity in the Calumet area, finding a PAH-free control would be a challenge.

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APPENDIX A

16S rDNA DATA

DNA Filter Quantification and NanoDrop Data for 16S rDNA Analysis Nanodrop data biomass/ total sediment filter weight Quarter percentage weight (-0.228) weight quarter of the site sample 260/280 ng/ul (g) (g) (g) sum (g) filter creosote 1A 1.48 6.7 0.048 22.86 marsh 1b 1.36 7.3 0.041 19.52 1c 1.72 7.8 0.05 23.81 freezer 0.304 0.007 0.071 0.21 33.81 big 2a 1.95 3 0.268 36.31 marsh 2b 2.31 3 0.102 13.82 well 2c 1.19 4.2 0.175 23.71 freezer 0.524 0.227 0.193 0.738 26.15 slag 3a 1.56 9.8 0.094 18.08 dune 3b 1.18 3.6 0.126 24.23 well #3 3c 1.39 11.3 0.201 38.65 freezer 0.34 0.043 0.099 0.52 19.04 big 4a 1.94 6.4 0.141 25.09 marsh 4b 1.75 6 0.117 20.82 surface 4c 2.07 6.6 0.149 26.51 water freezer 0.393 0.096 0.155 0.562 27.58 Railroa 5a 1.68 13 0.157 26.98 d ditch 5b 1.82 10.8 0.131 22.51 5c 1.62 16.8 0.129 22.16 freezer 0.458 0.161 0.165 0.582 28.35

16S rDNA Phylum Data OTUs Big Big Creosote Marsh Slag Dune Marsh Phylum Marsh Well Well #3 SW RR Ditch SW Archaea-Euryarchaeota 22 1 1 1 35 Archaea-Thaumarchaeota 3 26 0 2 0 Archaea-Undefined 7 0 0 0 10 Bacteria-Acidobacteria 105 375 50 28 31 Bacteria-Actinobacteria 96530 39615 4725 34 26240 Bacteria-Armatimonadetes 48 14 2 5 89 Bacteria-BRC1 0 0 1 0 0 Bacteria-Bacteroidetes 32767 1143 623 479 15804 Bacteria-Chlamydiae 96 34 0 7 6 Bacteria-Chlorobi 1 0 0 0 0 Bacteria-Chloroflexi 101 140 441 5 42 Bacteria-Cloacimonetes 1 0 0 1 0 Bacteria-Cyanobacteria-Chloroplast 800 390 57 7808 1239 Bacteria-Deinococcus-Thermus 14 127 63 3 6 Bacteria-Fibrobacteres 0 0 0 0 1 Bacteria-Firmicutes 439 22091 53163 293 260 Bacteria-Fusobacteria 24 3 0 0 110 Bacteria-Gemmatimonadetes 18 37 5 0 2 Bacteria-Hydrogenedentes 0 1 0 0 0 Bacteria-Ignavibacteriae 3 3 0 0 3 Bacteria-Latescibacteria 1 0 1 0 1 Bacteria-Lentisphaerae 3 0 0 0 3 Bacteria-Microgenomates 1 0 0 0 14 Bacteria-Nitrospirae 3 114 1 0 1 Bacteria-Parcubacteria 304 2 0 2 154 Bacteria-Planctomycetes 222 132 13 22 25 Bacteria-Proteobacteria 43949 52086 47209 79561 40736 Bacteria-SR1 23 1 0 0 4 Bacteria-Spirochaetes 3 0 0 0 4 Bacteria-Synergistetes 0 0 3 0 0 Bacteria-Tenericutes 0 0 3 0 0 Bacteria-Undefined 1652 842 2207 5457 916 Bacteria-Verrucomicrobia 1087 154 13 76 125 Below_Threshold-Archaea 17 8 3 157 54

16S rDNA Class Data OTUs Big Big RR Creosote Marsh Slag Dune Marsh Ditch Class Marsh Well Well #3 SW SW Archaea-Euryarchaeota-Archaeoglobi 1 0 0 0 0 Archaea-Euryarchaeota-Methanobacteria 1 0 0 0 1 Archaea-Euryarchaeota-Methanomicrobia 2 0 0 0 2 Archaea-Euryarchaeota-Thermoplasmata 0 0 1 1 0 Archaea-Euryarchaeota-Undefined 18 1 0 0 32 Archaea-Thaumarchaeota-Nitrosopumilales 0 2 0 0 0 Archaea-Thaumarchaeota-Nitrososphaerales 3 24 0 2 0 Archaea-Undefined 7 0 0 0 10 Bacteria-Acidobacteria-Acidobacteria_Gp1 9 2 3 8 4 Bacteria-Acidobacteria-Acidobacteria_Gp10 5 2 1 0 1 Bacteria-Acidobacteria-Acidobacteria_Gp13 0 1 0 0 0 Bacteria-Acidobacteria-Acidobacteria_Gp16 10 54 4 2 3 Bacteria-Acidobacteria-Acidobacteria_Gp17 1 7 1 0 3 Bacteria-Acidobacteria-Acidobacteria_Gp2 4 0 0 5 1 Bacteria-Acidobacteria-Acidobacteria_Gp25 0 0 1 0 0 Bacteria-Acidobacteria-Acidobacteria_Gp3 29 40 4 10 1 Bacteria-Acidobacteria-Acidobacteria_Gp4 13 177 19 0 4 Bacteria-Acidobacteria-Acidobacteria_Gp5 1 0 1 0 0 Bacteria-Acidobacteria-Acidobacteria_Gp6 24 71 13 2 10 Bacteria-Acidobacteria-Acidobacteria_Gp7 3 19 3 1 0 Bacteria-Acidobacteria-Holophagae 4 0 0 0 4 Bacteria-Acidobacteria-Undefined 2 2 0 0 0 Bacteria-Actinobacteria-Actinobacteria 96529 39583 4724 33 26238 Bacteria-Actinobacteria-Thermoleophilia 0 15 0 1 1 Bacteria-Actinobacteria-Undefined 1 17 1 0 1 Bacteria-Armatimonadetes- Armatimonadetes_gp2 3 0 2 0 0 Bacteria-Armatimonadetes- Armatimonadetes_gp5 6 9 0 5 1 Bacteria-Armatimonadetes-Armatimonadia 37 5 0 0 88 Bacteria-Armatimonadetes-Undefined 2 0 0 0 0 Bacteria-BRC1- BRC1_genera_incertae_sedis 0 0 1 0 0 Bacteria-Bacteroidetes- Bacteroidetes_incertae_sedis 47 5 0 1 3 Bacteria-Bacteroidetes-Bacteroidia 59 7 34 4 57 Bacteria-Bacteroidetes-Cytophagia 7652 160 12 259 2887 Bacteria-Bacteroidetes-Flavobacteriia 7477 341 328 114 5854 Bacteria-Bacteroidetes-Sphingobacteriia 16976 591 117 70 5969 54

Bacteria-Bacteroidetes-Undefined 556 39 132 31 1034 Bacteria-Chlamydiae-Chlamydiia 96 34 0 7 6 Bacteria-Chlorobi-Chlorobia 1 0 0 0 0 Bacteria-Chloroflexi-Anaerolineae 21 11 2 1 30 Bacteria-Chloroflexi-Caldilineae 6 3 1 0 4 Bacteria-Chloroflexi-Chloroflexia 26 13 1 0 0 Bacteria-Chloroflexi-Dehalococcoidetes 1 0 0 0 0 Bacteria-Chloroflexi-Ktedonobacteria 1 2 1 3 1 Bacteria-Chloroflexi-Thermomicrobia 0 71 376 1 0 Bacteria-Chloroflexi-Undefined 46 40 60 0 7 Bacteria-Cloacimonetes-Undefined 1 0 0 1 0 Bacteria-Cyanobacteria-Chloroplast- Chloroplast 500 262 33 7737 1079 Bacteria-Cyanobacteria-Chloroplast- Cyanobacteria 187 127 24 7 133 Bacteria-Cyanobacteria-Chloroplast- Undefined 113 1 0 64 27 Bacteria-Deinococcus-Thermus-Deinococci 14 127 63 3 6 Bacteria-Fibrobacteres-Fibrobacteria 0 0 0 0 1 Bacteria-Firmicutes-Bacilli 123 21963 1073 82 42 Bacteria-Firmicutes-Clostridia 282 104 7217 170 193 Bacteria-Firmicutes-Erysipelotrichia 3 3 41 3 5 Bacteria-Firmicutes-Negativicutes 6 0 2 4 9 Bacteria-Firmicutes-Undefined 25 21 44830 34 11 Bacteria-Fusobacteria-Fusobacteriia 24 3 0 0 110 Bacteria-Gemmatimonadetes- Gemmatimonadetes 18 37 5 0 2 Bacteria-Hydrogenedentes-Undefined 0 1 0 0 0 Bacteria-Ignavibacteriae-Ignavibacteria 3 3 0 0 3 Bacteria-Latescibacteria- Latescibacteria_genera_incertae_sedis 1 0 1 0 1 Bacteria-Lentisphaerae-Lentisphaeria 2 0 0 0 2 Bacteria-Lentisphaerae-Oligosphaeria 1 0 0 0 1 Bacteria-Microgenomates- Microgenomates_genera_incertae_sedis 1 0 0 0 14 Bacteria-Nitrospirae-Nitrospira 3 114 1 0 1 Bacteria-Parcubacteria- Parcubacteria_genera_incertae_sedis 304 2 0 2 154 Bacteria-Planctomycetes-Planctomycetia 222 132 13 22 25 Bacteria-Proteobacteria-Alphaproteobacteria 3554 22193 5615 11978 3087 Bacteria-Proteobacteria-Betaproteobacteria 37007 23395 37115 37845 34587 Bacteria-Proteobacteria-Deltaproteobacteria 284 300 35 14 114 Bacteria-Proteobacteria- Epsilonproteobacteria 50 0 0 1 258 Bacteria-Proteobacteria- Gammaproteobacteria 2444 6064 4340 29528 2361 55

Bacteria-Proteobacteria-Undefined 610 134 104 195 329 Bacteria-SR1-SR1_genera_incertae_sedis 23 1 0 0 4 Bacteria- Saccharibacteria_genera_incertae_sedis 0 1 0 1 1 Bacteria-Spirochaetes-Spirochaetia 3 0 0 0 4 Bacteria-Synergistetes-Synergistia 0 0 3 0 0 Bacteria-Tenericutes-Mollicutes 0 0 3 0 0 Bacteria-Undefined 1630 827 2197 5454 914 Bacteria-Verrucomicrobia-Opitutae 364 120 2 2 44 Bacteria-Verrucomicrobia-Spartobacteria 482 13 4 72 8 Bacteria-Verrucomicrobia-Subdivision3 30 8 1 1 11 Bacteria-Verrucomicrobia-Subdivision5 6 0 0 0 2 Bacteria-Verrucomicrobia-Undefined 24 4 0 1 2 Bacteria-Verrucomicrobia-Verrucomicrobiae 181 9 6 0 58 Bacteria-WPS-1_genera_incertae_sedis 3 3 2 1 1 Bacteria-WPS-2_genera_incertae_sedis 19 11 8 1 0 Below_Threshold-Archaea 17 8 3 157 54

16S rDNA Order Data OTUs Slag Big Dune Big Creosote Marsh Well Marsh RR Ditch Order Marsh Well #3 SW SW Archaea-Euryarchaeota-Archaeoglobi- Archaeoglobales 1 0 0 0 0 Archaea-Euryarchaeota-Methanobacteria- Methanobacteriales 1 0 0 0 1 Archaea-Euryarchaeota-Methanomicrobia- Methanomicrobiales 2 0 0 0 0 Archaea-Euryarchaeota-Methanomicrobia- Undefined 0 0 0 0 2 Archaea-Euryarchaeota-Thermoplasmata- Thermoplasmatales 0 0 1 1 0 Archaea-Euryarchaeota-Undefined 18 1 0 0 32 Archaea-Thaumarchaeota-Nitrosopumilales- Nitrosopumilales 0 2 0 0 0 Archaea-Thaumarchaeota-Nitrososphaerales- Nitrososphaerales 3 24 0 2 0 Archaea-Undefined 7 0 0 0 10 Bacteria-Acidobacteria-Acidobacteria_Gp1- Gp1 6 0 2 8 3 Bacteria-Acidobacteria-Acidobacteria_Gp1- Undefined 3 2 1 0 1 Bacteria-Acidobacteria-Acidobacteria_Gp10- Gp10 5 2 1 0 1 Bacteria-Acidobacteria-Acidobacteria_Gp13- Gp13 0 1 0 0 0 Bacteria-Acidobacteria-Acidobacteria_Gp16- Gp16 10 54 4 2 3 Bacteria-Acidobacteria-Acidobacteria_Gp17- Gp17 1 7 1 0 3 Bacteria-Acidobacteria-Acidobacteria_Gp2- Gp2 4 0 0 5 1 Bacteria-Acidobacteria-Acidobacteria_Gp25- Gp25 0 0 1 0 0 Bacteria-Acidobacteria-Acidobacteria_Gp3- Bryobacter 4 0 0 0 0 Bacteria-Acidobacteria-Acidobacteria_Gp3- Gp3 22 38 4 9 0 Bacteria-Acidobacteria-Acidobacteria_Gp3- Undefined 3 2 0 1 1 Bacteria-Acidobacteria-Acidobacteria_Gp4- Blastocatella 5 8 12 0 1 Bacteria-Acidobacteria-Acidobacteria_Gp4- Gp4 8 157 7 0 3 Bacteria-Acidobacteria-Acidobacteria_Gp4- Undefined 0 12 0 0 0 Bacteria-Acidobacteria-Acidobacteria_Gp5- Gp5 1 0 1 0 0 57

Bacteria-Acidobacteria-Acidobacteria_Gp6- Gp6 24 71 13 2 10 Bacteria-Acidobacteria-Acidobacteria_Gp7- Gp7 3 19 3 1 0 Bacteria-Acidobacteria-Holophagae- Holophagales 4 0 0 0 4 Bacteria-Acidobacteria-Undefined 2 2 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Acidimicrobiales 20 222 56 0 41 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales 96423 38226 4311 21 26120 Bacteria-Actinobacteria-Actinobacteria- Coriobacteriales 6 22 5 0 2 Bacteria-Actinobacteria-Actinobacteria- Gaiellales 8 77 5 6 0 Bacteria-Actinobacteria-Actinobacteria- Nitriliruptorales 0 4 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Rubrobacterales 0 26 3 0 0 Bacteria-Actinobacteria-Actinobacteria- Solirubrobacterales 10 916 325 4 3 Bacteria-Actinobacteria-Actinobacteria- Undefined 62 90 19 2 72 Bacteria-Actinobacteria-Thermoleophilia- Thermoleophilales 0 15 0 1 1 Bacteria-Actinobacteria-Undefined 1 17 1 0 1 Bacteria-Armatimonadetes- Armatimonadetes_gp2-Armatimonadetes_gp2 3 0 2 0 0 Bacteria-Armatimonadetes- Armatimonadetes_gp5-Armatimonadetes_gp5 6 9 0 5 1 Bacteria-Armatimonadetes-Armatimonadia- Armatimonadales 37 5 0 0 88 Bacteria-Armatimonadetes-Undefined 2 0 0 0 0 Bacteria-BRC1-BRC1_genera_incertae_sedis- BRC1_genera_incertae_sedis 0 0 1 0 0 Bacteria-Bacteroidetes- Bacteroidetes_incertae_sedis-Ohtaekwangia 47 5 0 1 3 Bacteria-Bacteroidetes-Bacteroidia- Bacteroidales 59 7 34 4 57 Bacteria-Bacteroidetes-Cytophagia- Cytophagales 7652 160 12 259 2887 Bacteria-Bacteroidetes-Flavobacteriia- Flavobacteriales 7477 341 328 114 5854 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales 16976 591 117 70 5969 Bacteria-Bacteroidetes-Undefined 556 39 132 31 1034 Bacteria-Chlamydiae-Chlamydiia- Chlamydiales 96 34 0 7 6 Bacteria-Chlorobi-Chlorobia-Chlorobiales 1 0 0 0 0 Bacteria-Chloroflexi-Anaerolineae- Anaerolineales 21 11 2 1 30 Bacteria-Chloroflexi-Caldilineae-Caldilineales 6 3 1 0 4 58

Bacteria-Chloroflexi-Chloroflexia- Chloroflexales 26 13 1 0 0 Bacteria-Chloroflexi-Dehalococcoidetes- Dehalogenimonas 1 0 0 0 0 Bacteria-Chloroflexi-Ktedonobacteria- Ktedonobacterales 1 1 1 3 1 Bacteria-Chloroflexi-Ktedonobacteria- Undefined 0 1 0 0 0 Bacteria-Chloroflexi-Thermomicrobia- Sphaerobacterales 0 63 356 1 0 Bacteria-Chloroflexi-Thermomicrobia- Undefined 0 8 20 0 0 Bacteria-Chloroflexi-Undefined 46 40 60 0 7 Bacteria-Cloacimonetes-Undefined 1 0 0 1 0 Bacteria-Cyanobacteria-Chloroplast- Chloroplast-Chloroplast 500 262 33 7737 1079 Bacteria-Cyanobacteria-Chloroplast- Cyanobacteria-Undefined 187 127 24 7 133 Bacteria-Cyanobacteria-Chloroplast- Undefined 113 1 0 64 27 Bacteria-Deinococcus-Thermus-Deinococci- Deinococcales 14 121 23 2 6 Bacteria-Deinococcus-Thermus-Deinococci- Thermales 0 6 40 1 0 Bacteria-Fibrobacteres-Fibrobacteria- Fibrobacterales 0 0 0 0 1 Bacteria-Firmicutes-Bacilli-Bacillales 113 21915 1056 28 25 Bacteria-Firmicutes-Bacilli-Lactobacillales 9 32 14 53 17 Bacteria-Firmicutes-Bacilli-Undefined 1 16 3 1 0 Bacteria-Firmicutes-Clostridia-Clostridiales 277 97 3460 156 189 Bacteria-Firmicutes-Clostridia- Halanaerobiales 0 0 0 1 0 Bacteria-Firmicutes-Clostridia- Natranaerobiales 1 0 445 4 0 Bacteria-Firmicutes-Clostridia- Thermoanaerobacterales 0 0 2 0 0 Bacteria-Firmicutes-Clostridia-Undefined 4 7 3310 9 4 Bacteria-Firmicutes-Erysipelotrichia- Erysipelotrichales 3 3 41 3 5 Bacteria-Firmicutes-Negativicutes- Selenomonadales 6 0 2 4 9 Bacteria-Firmicutes-Undefined 25 21 44830 34 11 Bacteria-Fusobacteria-Fusobacteriia- Fusobacteriales 24 3 0 0 110 Bacteria-Gemmatimonadetes- Gemmatimonadetes-Gemmatimonadales 18 37 5 0 2 Bacteria-Hydrogenedentes-Undefined 0 1 0 0 0 Bacteria-Ignavibacteriae-Ignavibacteria- Ignavibacteriales 3 3 0 0 3 59

Bacteria-Latescibacteria- Latescibacteria_genera_incertae_sedis- Latescibacteria_genera_incertae_sedis 1 0 1 0 1 Bacteria-Lentisphaerae-Lentisphaeria- Victivallales 2 0 0 0 2 Bacteria-Lentisphaerae-Oligosphaeria- Oligosphaerales 1 0 0 0 1 Bacteria-Microgenomates- Microgenomates_genera_incertae_sedis- Microgenomates_genera_incertae_sedis 1 0 0 0 14 Bacteria-Nitrospirae-Nitrospira-Nitrospirales 3 114 1 0 1 Bacteria-Parcubacteria- Parcubacteria_genera_incertae_sedis- Parcubacteria_genera_incertae_sedis 304 2 0 2 154 Bacteria-Planctomycetes-Planctomycetia- Planctomycetales 220 131 13 22 24 Bacteria-Planctomycetes-Planctomycetia- Undefined 2 1 0 0 1 Bacteria-Proteobacteria-Alphaproteobacteria- Alphaproteobacteria_incertae_sedis 5 9 1 0 1 Bacteria-Proteobacteria-Alphaproteobacteria- Caulobacterales 292 15092 4034 5 43 Bacteria-Proteobacteria-Alphaproteobacteria- Parvularculales 3 0 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales 616 3333 614 16 242 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodobacterales 999 192 25 6425 1079 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales 199 402 704 3315 55 Bacteria-Proteobacteria-Alphaproteobacteria- Rickettsiales 53 2 5 1 41 Bacteria-Proteobacteria-Alphaproteobacteria- SAR11 0 0 0 0 28 Bacteria-Proteobacteria-Alphaproteobacteria- Sphingomonadales 927 3074 186 2188 1500 Bacteria-Proteobacteria-Alphaproteobacteria- Undefined 460 89 46 28 98 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales 33879 22873 36719 37726 30229 Bacteria-Proteobacteria-Betaproteobacteria- Ferrovales 2 0 0 0 1 Bacteria-Proteobacteria-Betaproteobacteria- Gallionellales 24 84 1 1 17 Bacteria-Proteobacteria-Betaproteobacteria- Hydrogenophilales 10 0 1 0 31 Bacteria-Proteobacteria-Betaproteobacteria- Methylophilales 186 11 1 4 789 Bacteria-Proteobacteria-Betaproteobacteria- Neisseriales 295 1 1 0 2085 Bacteria-Proteobacteria-Betaproteobacteria- Nitrosomonadales 6 17 0 0 2 60

Bacteria-Proteobacteria-Betaproteobacteria- Procabacteriales 0 1 0 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Rhodocyclales 206 304 335 48 289 Bacteria-Proteobacteria-Betaproteobacteria- Undefined 2399 104 57 66 1144 Bacteria-Proteobacteria-Deltaproteobacteria- Bdellovibrionales 73 26 6 3 33 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfobacterales 24 2 0 0 20 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfovibrionales 8 0 3 0 4 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfuromonadales 43 44 0 0 12 Bacteria-Proteobacteria-Deltaproteobacteria- Myxococcales 53 178 21 7 18 Bacteria-Proteobacteria-Deltaproteobacteria- Syntrophobacterales 23 23 0 1 8 Bacteria-Proteobacteria-Deltaproteobacteria- Syntrophorhabdaceae 0 0 0 0 1 Bacteria-Proteobacteria-Deltaproteobacteria- Undefined 60 27 5 3 18 Bacteria-Proteobacteria- Epsilonproteobacteria-Campylobacterales 29 0 0 1 257 Bacteria-Proteobacteria- Epsilonproteobacteria-Nautiliales 0 0 0 0 1 Bacteria-Proteobacteria- Epsilonproteobacteria-Undefined 21 0 0 0 0 Bacteria-Proteobacteria- Gammaproteobacteria-Aeromonadales 112 11 4 6 553 Bacteria-Proteobacteria- Gammaproteobacteria-Alteromonadales 37 0 0 0 14 Bacteria-Proteobacteria- Gammaproteobacteria-Chromatiales 467 28 3 3 505 Bacteria-Proteobacteria- Gammaproteobacteria-Enterobacteriales 102 3444 233 8 266 Bacteria-Proteobacteria- Gammaproteobacteria- Gammaproteobacteria_incertae_sedis 21 36 0 56 6 Bacteria-Proteobacteria- Gammaproteobacteria-Legionellales 656 104 38 22 26 Bacteria-Proteobacteria- Gammaproteobacteria-Methylococcales 24 6 3 5 22 Bacteria-Proteobacteria- Gammaproteobacteria-Oceanospirillales 7 11 0 1 4 Bacteria-Proteobacteria- Gammaproteobacteria-Orbales 1 0 0 0 0 Bacteria-Proteobacteria- Gammaproteobacteria-Pseudomonadales 74 1549 465 17 716 Bacteria-Proteobacteria- Gammaproteobacteria-Thiotrichales 9 1 0 0 3 Bacteria-Proteobacteria- Gammaproteobacteria-Undefined 672 100 57 69 197 61

Bacteria-Proteobacteria- Gammaproteobacteria-Vibrionales 1 0 0 0 0 Bacteria-Proteobacteria- Gammaproteobacteria-Xanthomonadales 261 774 3537 29341 49 Bacteria-Proteobacteria-Undefined 610 134 104 195 329 Bacteria-SR1-SR1_genera_incertae_sedis- SR1_genera_incertae_sedis 23 1 0 0 4 Bacteria- Saccharibacteria_genera_incertae_sedis- Saccharibacteria_genera_incertae_sedis 0 1 0 1 1 Bacteria-Spirochaetes-Spirochaetia- Spirochaetales 3 0 0 0 4 Bacteria-Synergistetes-Synergistia- Synergistales 0 0 3 0 0 Bacteria-Tenericutes-Mollicutes- Acholeplasmatales 0 0 3 0 0 Bacteria-Undefined 1630 827 2197 5454 914 Bacteria-Verrucomicrobia-Opitutae-Opitutales 355 119 2 1 6 Bacteria-Verrucomicrobia-Opitutae- Puniceicoccales 3 0 0 1 38 Bacteria-Verrucomicrobia-Opitutae- Undefined 6 1 0 0 0 Bacteria-Verrucomicrobia-Spartobacteria- Spartobacteria_genera_incertae_sedis 478 11 4 70 8 Bacteria-Verrucomicrobia-Spartobacteria- Undefined 3 1 0 2 0 Bacteria-Verrucomicrobia-Spartobacteria- Xiphinematobacter 1 1 0 0 0 Bacteria-Verrucomicrobia-Subdivision3- Subdivision3_genera_incertae_sedis 30 8 1 1 11 Bacteria-Verrucomicrobia-Subdivision5- Subdivision5_genera_incertae_sedis 6 0 0 0 2 Bacteria-Verrucomicrobia-Undefined 24 4 0 1 2 Bacteria-Verrucomicrobia-Verrucomicrobiae- Verrucomicrobiales 181 9 6 0 58 Bacteria-WPS-1_genera_incertae_sedis-WPS- 1_genera_incertae_sedis 3 3 2 1 1 Bacteria-WPS-2_genera_incertae_sedis-WPS- 2_genera_incertae_sedis 19 11 8 1 0 Below_Threshold-Archaea 17 8 3 157 54

16S rDNA Family Data OTUs Slag Big Dune Big Creosote Marsh Well Marsh Family Marsh Well #3 SW RR Ditch SW Archaea-Euryarchaeota-Archaeoglobi- Archaeoglobales-Archaeoglobaceae 1 0 0 0 0 Archaea-Euryarchaeota-Methanobacteria- Methanobacteriales-Methanobacteriaceae 1 0 0 0 1 Archaea-Euryarchaeota-Methanomicrobia- Methanomicrobiales- Methanomicrobiales_incertae_sedis 1 0 0 0 0 Archaea-Euryarchaeota-Methanomicrobia- Methanomicrobiales-Methanospirillaceae 1 0 0 0 0 Archaea-Euryarchaeota-Methanomicrobia- Undefined 0 0 0 0 2 Archaea-Euryarchaeota-Thermoplasmata- Thermoplasmatales-Undefined 0 0 1 1 0 Archaea-Euryarchaeota-Undefined 18 1 0 0 32 Archaea-Thaumarchaeota-Nitrosopumilales- Nitrosopumilales-Nitrosopumilaceae 0 2 0 0 0 Archaea-Thaumarchaeota-Nitrososphaerales- Nitrososphaerales-Nitrososphaeraceae 3 24 0 2 0 Archaea-Undefined 7 0 0 0 10 Bacteria-Acidobacteria-Acidobacteria_Gp1- Gp1-Gp1 6 0 2 8 3 Bacteria-Acidobacteria-Acidobacteria_Gp1- Undefined 3 2 1 0 1 Bacteria-Acidobacteria-Acidobacteria_Gp10- Gp10-Gp10 5 2 1 0 1 Bacteria-Acidobacteria-Acidobacteria_Gp13- Gp13-Gp13 0 1 0 0 0 Bacteria-Acidobacteria-Acidobacteria_Gp16- Gp16-Gp16 10 54 4 2 3 Bacteria-Acidobacteria-Acidobacteria_Gp17- Gp17-Gp17 1 7 1 0 3 Bacteria-Acidobacteria-Acidobacteria_Gp2- Gp2-Gp2 4 0 0 5 1 Bacteria-Acidobacteria-Acidobacteria_Gp25- Gp25-Gp25 0 0 1 0 0 Bacteria-Acidobacteria-Acidobacteria_Gp3- Bryobacter-Bryobacter 4 0 0 0 0 Bacteria-Acidobacteria-Acidobacteria_Gp3- Gp3-Gp3 22 38 4 9 0 Bacteria-Acidobacteria-Acidobacteria_Gp3- Undefined 3 2 0 1 1 Bacteria-Acidobacteria-Acidobacteria_Gp4- Blastocatella-Blastocatella 5 8 12 0 1 Bacteria-Acidobacteria-Acidobacteria_Gp4- Gp4-Gp4 8 157 7 0 3 63

Bacteria-Acidobacteria-Acidobacteria_Gp4- Undefined 0 12 0 0 0 Bacteria-Acidobacteria-Acidobacteria_Gp5- Gp5-Gp5 1 0 1 0 0 Bacteria-Acidobacteria-Acidobacteria_Gp6- Gp6-Gp6 24 71 13 2 10 Bacteria-Acidobacteria-Acidobacteria_Gp7- Gp7-Gp7 3 19 3 1 0 Bacteria-Acidobacteria-Holophagae- Holophagales-Holophagaceae 4 0 0 0 4 Bacteria-Acidobacteria-Undefined 2 2 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Acidimicrobiales-Acidimicrobiaceae 7 14 8 0 31 Bacteria-Actinobacteria-Actinobacteria- Acidimicrobiales- Acidimicrobineae_incertae_sedis 5 93 3 0 0 Bacteria-Actinobacteria-Actinobacteria- Acidimicrobiales-Iamiaceae 0 49 18 0 0 Bacteria-Actinobacteria-Actinobacteria- Acidimicrobiales-Undefined 8 66 27 0 10 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Acidothermaceae 0 2 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Beutenbergiaceae 0 1 2 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Bogoriellaceae 0 81 27 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Brevibacteriaceae 0 45 101 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Cellulomonadaceae 0 189 14 0 1 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Corynebacteriaceae 1 16 13 0 1 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Cryptosporangiaceae 0 11 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Demequinaceae 17 11 0 0 4 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Dermabacteraceae 0 143 140 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Dermacoccaceae 0 0 1 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Dermatophilaceae 2 0 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Dietziaceae 0 6131 960 1 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Geodermatophilaceae 2330 5563 271 0 1401 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Intrasporangiaceae 10448 62 26 0 1266 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Jiangellaceae 0 0 1 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Kineosporiaceae 1 174 18 0 7 64

Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Microbacteriaceae 43644 975 546 4 6576 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Micrococcaceae 3 5845 361 4 1 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales- Micrococcineae_incertae_sedis 0 1 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Micromonosporaceae 4 33 13 0 2 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Mycobacteriaceae 158 332 13 1 5 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Nakamurellaceae 0 26 7 0 1 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Nocardiaceae 1 3162 385 0 1 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Nocardioidaceae 10 1381 210 3 7 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Nocardiopsaceae 0 12 9 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Promicromonosporaceae 4 76 49 6 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Propionibacteriaceae 1 0 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Pseudonocardiaceae 1 1227 985 1 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Ruaniaceae 0 10 2 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Sanguibacteraceae 1 11535 1 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Sporichthyaceae 4 0 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Streptomycetaceae 1 54 92 1 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Streptosporangiaceae 0 2 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Thermomonosporaceae 2 4 1 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Tsukamurellaceae 0 3 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Undefined 39790 1119 63 0 16847 Bacteria-Actinobacteria-Actinobacteria- Coriobacteriales-Coriobacteriaceae 6 22 5 0 2 Bacteria-Actinobacteria-Actinobacteria- Gaiellales-Gaiellaceae 8 77 5 6 0 Bacteria-Actinobacteria-Actinobacteria- Nitriliruptorales-Nitriliruptoraceae 0 4 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Rubrobacterales-Rubrobacteraceae 0 26 3 0 0 Bacteria-Actinobacteria-Actinobacteria- Solirubrobacterales-Conexibacteraceae 5 579 13 0 0 Bacteria-Actinobacteria-Actinobacteria- Solirubrobacterales-Patulibacteraceae 0 143 273 0 0 65

Bacteria-Actinobacteria-Actinobacteria- Solirubrobacterales-Solirubrobacteraceae 3 93 20 3 0 Bacteria-Actinobacteria-Actinobacteria- Solirubrobacterales-Undefined 2 101 19 1 3 Bacteria-Actinobacteria-Actinobacteria- Undefined 62 90 19 2 72 Bacteria-Actinobacteria-Thermoleophilia- Thermoleophilales-Thermoleophilaceae 0 15 0 1 1 Bacteria-Actinobacteria-Undefined 1 17 1 0 1 Bacteria-Armatimonadetes- Armatimonadetes_gp2- Armatimonadetes_gp2-Armatimonadetes_gp2 3 0 2 0 0 Bacteria-Armatimonadetes- Armatimonadetes_gp5- Armatimonadetes_gp5-Armatimonadetes_gp5 6 9 0 5 1 Bacteria-Armatimonadetes-Armatimonadia- Armatimonadales-Armatimonadaceae 37 5 0 0 88 Bacteria-Armatimonadetes-Undefined 2 0 0 0 0 Bacteria-BRC1- BRC1_genera_incertae_sedis- BRC1_genera_incertae_sedis- BRC1_genera_incertae_sedis 0 0 1 0 0 Bacteria-Bacteroidetes- Bacteroidetes_incertae_sedis-Ohtaekwangia- Ohtaekwangia 47 5 0 1 3 Bacteria-Bacteroidetes-Bacteroidia- Bacteroidales-Bacteroidaceae 11 0 20 1 16 Bacteria-Bacteroidetes-Bacteroidia- Bacteroidales-Marinilabiliaceae 0 0 0 0 2 Bacteria-Bacteroidetes-Bacteroidia- Bacteroidales-Porphyromonadaceae 32 6 11 2 25 Bacteria-Bacteroidetes-Bacteroidia- Bacteroidales-Prevotellaceae 0 0 0 0 7 Bacteria-Bacteroidetes-Bacteroidia- Bacteroidales-Sunxiuqinia 4 0 0 0 0 Bacteria-Bacteroidetes-Bacteroidia- Bacteroidales-Undefined 12 1 3 1 7 Bacteria-Bacteroidetes-Cytophagia- Cytophagales-Cytophagaceae 7564 147 12 87 2733 Bacteria-Bacteroidetes-Cytophagia- Cytophagales-Flammeovirgaceae 30 8 0 144 90 Bacteria-Bacteroidetes-Cytophagia- Cytophagales-Mooreiaceae 0 1 0 0 0 Bacteria-Bacteroidetes-Cytophagia- Cytophagales-Undefined 58 4 0 28 64 Bacteria-Bacteroidetes-Flavobacteriia- Flavobacteriales-Cryomorphaceae 1312 12 105 58 1583 Bacteria-Bacteroidetes-Flavobacteriia- Flavobacteriales-Flavobacteriaceae 6124 326 200 18 4129 Bacteria-Bacteroidetes-Flavobacteriia- Flavobacteriales-Undefined 41 3 23 38 142 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Chitinophagaceae 16477 312 10 52 5583 66

Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Cyclobacteriaceae 30 14 2 15 54 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Rhodothermaceae 2 2 2 0 0 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Saprospiraceae 349 12 0 0 90 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Sphingobacteriaceae 35 240 101 1 23 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Undefined 83 11 2 2 219 Bacteria-Bacteroidetes-Undefined 556 39 132 31 1034 Bacteria-Chlamydiae-Chlamydiia- Chlamydiales-Chlamydiaceae 2 0 0 0 0 Bacteria-Chlamydiae-Chlamydiia- Chlamydiales-Parachlamydiaceae 49 14 0 2 1 Bacteria-Chlamydiae-Chlamydiia- Chlamydiales-Simkaniaceae 26 7 0 2 2 Bacteria-Chlamydiae-Chlamydiia- Chlamydiales-Undefined 19 13 0 3 3 Bacteria-Chlorobi-Chlorobia-Chlorobiales- Chlorobiaceae 1 0 0 0 0 Bacteria-Chloroflexi-Anaerolineae- Anaerolineales-Anaerolineaceae 21 11 2 1 30 Bacteria-Chloroflexi-Caldilineae- Caldilineales-Caldilineaceae 6 3 1 0 4 Bacteria-Chloroflexi-Chloroflexia- Chloroflexales-Chloroflexaceae 1 3 1 0 0 Bacteria-Chloroflexi-Chloroflexia- Chloroflexales-Oscillochloridaceae 24 5 0 0 0 Bacteria-Chloroflexi-Chloroflexia- Chloroflexales-Undefined 1 5 0 0 0 Bacteria-Chloroflexi-Dehalococcoidetes- Dehalogenimonas-Dehalogenimonas 1 0 0 0 0 Bacteria-Chloroflexi-Ktedonobacteria- Ktedonobacterales-Ktedonobacteraceae 1 1 1 3 1 Bacteria-Chloroflexi-Ktedonobacteria- Undefined 0 1 0 0 0 Bacteria-Chloroflexi-Thermomicrobia- Sphaerobacterales-Sphaerobacteraceae 0 63 356 1 0 Bacteria-Chloroflexi-Thermomicrobia- Undefined 0 8 20 0 0 Bacteria-Chloroflexi-Undefined 46 40 60 0 7 Bacteria-Cloacimonetes-Undefined 1 0 0 1 0 Bacteria-Cyanobacteria-Chloroplast- Chloroplast-Chloroplast-Chloroplast 500 262 33 7737 1079 Bacteria-Cyanobacteria-Chloroplast- Cyanobacteria-Undefined 187 127 24 7 133 Bacteria-Cyanobacteria-Chloroplast- Undefined 113 1 0 64 27 Bacteria-Deinococcus-Thermus-Deinococci- Deinococcales-Deinococcaceae 10 75 1 0 6 Bacteria-Deinococcus-Thermus-Deinococci- Deinococcales-Trueperaceae 4 46 22 2 0 67

Bacteria-Deinococcus-Thermus-Deinococci- Thermales-Thermaceae 0 6 40 1 0 Bacteria-Fibrobacteres-Fibrobacteria- Fibrobacterales-Fibrobacteraceae 0 0 0 0 1 Bacteria-Firmicutes-Bacilli-Bacillales- Alicyclobacillaceae 0 6 2 0 0 Bacteria-Firmicutes-Bacilli-Bacillales- Planococcaceae 10 12010 645 4 3 Bacteria-Firmicutes-Bacilli-Bacillales- Staphylococcaceae 0 324 28 0 0 Bacteria-Firmicutes-Bacilli-Bacillales- Undefined 103 9575 381 24 22 Bacteria-Firmicutes-Bacilli-Lactobacillales- Aerococcaceae 0 0 0 0 1 Bacteria-Firmicutes-Bacilli-Lactobacillales- Carnobacteriaceae 7 17 9 41 1 Bacteria-Firmicutes-Bacilli-Lactobacillales- Enterococcaceae 0 2 3 10 0 Bacteria-Firmicutes-Bacilli-Lactobacillales- Lactobacillaceae 0 0 0 0 1 Bacteria-Firmicutes-Bacilli-Lactobacillales- Leuconostocaceae 0 0 1 0 0 Bacteria-Firmicutes-Bacilli-Lactobacillales- Streptococcaceae 2 12 1 1 14 Bacteria-Firmicutes-Bacilli-Lactobacillales- Undefined 0 1 0 1 0 Bacteria-Firmicutes-Bacilli-Undefined 1 16 3 1 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Gracilibacteraceae 0 0 0 3 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Lachnospiraceae 22 11 0 16 10 Bacteria-Firmicutes-Clostridia-Clostridiales- Natranaerovirga 0 0 0 0 8 Bacteria-Firmicutes-Clostridia-Clostridiales- Peptostreptococcaceae 95 52 12 12 8 Bacteria-Firmicutes-Clostridia-Clostridiales- Ruminococcaceae 23 0 17 42 11 Bacteria-Firmicutes-Clostridia-Clostridiales- Syntrophomonadaceae 0 0 0 2 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Undefined 137 34 3431 81 152 Bacteria-Firmicutes-Clostridia- Halanaerobiales-Halobacteroidaceae 0 0 0 1 0 Bacteria-Firmicutes-Clostridia- Natranaerobiales-Natranaerobiaceae 1 0 445 4 0 Bacteria-Firmicutes-Clostridia- Thermoanaerobacterales- Thermoanaerobacteraceae 0 0 1 0 0 Bacteria-Firmicutes-Clostridia- Thermoanaerobacterales-Undefined 0 0 1 0 0 Bacteria-Firmicutes-Clostridia-Undefined 4 7 3310 9 4 Bacteria-Firmicutes-Erysipelotrichia- Erysipelotrichales-Erysipelotrichaceae 3 3 41 3 5 68

Bacteria-Firmicutes-Negativicutes- Selenomonadales-Acidaminococcaceae 0 0 0 0 1 Bacteria-Firmicutes-Negativicutes- Selenomonadales-Veillonellaceae 6 0 2 4 8 Bacteria-Firmicutes-Undefined 25 21 44830 34 11 Bacteria-Fusobacteria-Fusobacteriia- Fusobacteriales-Fusobacteriaceae 24 3 0 0 110 Bacteria-Gemmatimonadetes- Gemmatimonadetes-Gemmatimonadales- Gemmatimonadaceae 18 37 5 0 2 Bacteria-Hydrogenedentes-Undefined 0 1 0 0 0 Bacteria-Ignavibacteriae-Ignavibacteria- Ignavibacteriales-Ignavibacteriaceae 3 3 0 0 3 Bacteria-Latescibacteria- Latescibacteria_genera_incertae_sedis- Latescibacteria_genera_incertae_sedis- Latescibacteria_genera_incertae_sedis 1 0 1 0 1 Bacteria-Lentisphaerae-Lentisphaeria- Victivallales-Victivallaceae 2 0 0 0 2 Bacteria-Lentisphaerae-Oligosphaeria- Oligosphaerales-Oligosphaeraceae 1 0 0 0 1 Bacteria-Microgenomates- Microgenomates_genera_incertae_sedis- Microgenomates_genera_incertae_sedis- Microgenomates_genera_incertae_sedis 1 0 0 0 14 Bacteria-Nitrospirae-Nitrospira-Nitrospirales- Nitrospiraceae 3 114 1 0 1 Bacteria-Parcubacteria- Parcubacteria_genera_incertae_sedis- Parcubacteria_genera_incertae_sedis- Parcubacteria_genera_incertae_sedis 304 2 0 2 154 Bacteria-Planctomycetes-Planctomycetia- Planctomycetales-Planctomycetaceae 220 131 13 22 24 Bacteria-Planctomycetes-Planctomycetia- Undefined 2 1 0 0 1 Bacteria-Proteobacteria-Alphaproteobacteria- Alphaproteobacteria_incertae_sedis- Geminicoccus 4 7 1 0 1 Bacteria-Proteobacteria-Alphaproteobacteria- Alphaproteobacteria_incertae_sedis- Rhizomicrobium 1 2 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Caulobacterales-Caulobacteraceae 265 15090 4032 5 38 Bacteria-Proteobacteria-Alphaproteobacteria- Caulobacterales-Hyphomonadaceae 27 2 0 0 5 Bacteria-Proteobacteria-Alphaproteobacteria- Caulobacterales-Undefined 0 0 2 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Parvularculales-Parvularculaceae 3 0 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Aurantimonadaceae 2 37 9 0 2 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Beijerinckiaceae 12 53 2 4 93 69

Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Bradyrhizobiaceae 29 199 100 4 12 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Brucellaceae 0 23 18 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Hyphomicrobiaceae 144 881 185 2 20 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Methylobacteriaceae 17 1585 106 0 8 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Methylocystaceae 9 46 95 0 3 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Phyllobacteriaceae 23 135 52 1 4 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Rhizobiaceae 54 195 10 0 25 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Rhizobiales_incertae_sedis 4 7 2 0 2 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Rhodobiaceae 7 1 0 0 3 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Undefined 303 111 30 1 67 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Xanthobacteraceae 12 60 5 4 3 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodobacterales-Rhodobacteraceae 999 192 25 6425 1079 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Acetobacteraceae 110 333 701 3313 14 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Elioraea 1 0 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Rhodospirillaceae 69 54 0 0 36 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Undefined 19 15 3 2 5 Bacteria-Proteobacteria-Alphaproteobacteria- Rickettsiales-Anaplasmataceae 9 0 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rickettsiales-Rickettsiaceae 41 2 5 1 40 Bacteria-Proteobacteria-Alphaproteobacteria- Rickettsiales-Undefined 3 0 0 0 1 Bacteria-Proteobacteria-Alphaproteobacteria- SAR11-SAR11 0 0 0 0 28 Bacteria-Proteobacteria-Alphaproteobacteria- Sphingomonadales-Erythrobacteraceae 473 52 2 2119 48 Bacteria-Proteobacteria-Alphaproteobacteria- Sphingomonadales-Sphingomonadaceae 444 3017 182 58 1446 Bacteria-Proteobacteria-Alphaproteobacteria- Sphingomonadales-Undefined 10 5 2 11 6 Bacteria-Proteobacteria-Alphaproteobacteria- Undefined 460 89 46 28 98 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Alcaligenaceae 19 78 1819 1 388 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Burkholderiaceae 10597 19 11 0 9991 70

Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales- Burkholderiales_incertae_sedis 2040 63 82 63 1623 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae 16932 18140 34130 37149 15251 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Oxalobacteraceae 412 4346 342 2 584 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Sutterellaceae 1 0 0 0 6 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Undefined 3878 227 335 511 2386 Bacteria-Proteobacteria-Betaproteobacteria- Ferrovales-Ferrovaceae 2 0 0 0 1 Bacteria-Proteobacteria-Betaproteobacteria- Gallionellales-Gallionellaceae 24 84 1 1 17 Bacteria-Proteobacteria-Betaproteobacteria- Hydrogenophilales-Hydrogenophilaceae 10 0 1 0 31 Bacteria-Proteobacteria-Betaproteobacteria- Methylophilales-Methylophilaceae 186 11 1 4 789 Bacteria-Proteobacteria-Betaproteobacteria- Neisseriales-Neisseriaceae 295 1 1 0 2085 Bacteria-Proteobacteria-Betaproteobacteria- Nitrosomonadales-Nitrosomonadaceae 6 17 0 0 2 Bacteria-Proteobacteria-Betaproteobacteria- Procabacteriales-Procabacteriaceae 0 1 0 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Rhodocyclales-Rhodocyclaceae 206 304 335 48 289 Bacteria-Proteobacteria-Betaproteobacteria- Undefined 2399 104 57 66 1144 Bacteria-Proteobacteria-Deltaproteobacteria- Bdellovibrionales-Bacteriovoracaceae 27 3 6 0 15 Bacteria-Proteobacteria-Deltaproteobacteria- Bdellovibrionales-Bdellovibrionaceae 46 23 0 3 18 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfobacterales-Desulfobacteraceae 12 2 0 0 8 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfobacterales-Desulfobulbaceae 12 0 0 0 10 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfobacterales-Undefined 0 0 0 0 2 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfovibrionales-Desulfovibrionaceae 8 0 1 0 4 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfovibrionales-Undefined 0 0 2 0 0 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfuromonadales-Desulfuromonadaceae 2 0 0 0 0 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfuromonadales-Geobacteraceae 38 44 0 0 12 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfuromonadales-Undefined 3 0 0 0 0 Bacteria-Proteobacteria-Deltaproteobacteria- Myxococcales-Cystobacteraceae 10 22 1 0 5 Bacteria-Proteobacteria-Deltaproteobacteria- Myxococcales-Kofleriaceae 0 1 0 0 0 71

Bacteria-Proteobacteria-Deltaproteobacteria- Myxococcales-Myxococcaceae 0 6 3 1 0 Bacteria-Proteobacteria-Deltaproteobacteria- Myxococcales-Nannocystaceae 1 1 0 1 0 Bacteria-Proteobacteria-Deltaproteobacteria- Myxococcales-Phaselicystidaceae 3 0 0 0 0 Bacteria-Proteobacteria-Deltaproteobacteria- Myxococcales-Polyangiaceae 15 102 11 2 5 Bacteria-Proteobacteria-Deltaproteobacteria- Myxococcales-Undefined 24 46 6 3 8 Bacteria-Proteobacteria-Deltaproteobacteria- Syntrophobacterales-Syntrophaceae 16 12 0 1 5 Bacteria-Proteobacteria-Deltaproteobacteria- Syntrophobacterales-Syntrophobacteraceae 7 10 0 0 2 Bacteria-Proteobacteria-Deltaproteobacteria- Syntrophobacterales-Undefined 0 1 0 0 1 Bacteria-Proteobacteria-Deltaproteobacteria- Syntrophorhabdaceae-Syntrophorhabdaceae 0 0 0 0 1 Bacteria-Proteobacteria-Deltaproteobacteria- Undefined 60 27 5 3 18 Bacteria-Proteobacteria- Epsilonproteobacteria-Campylobacterales- Campylobacteraceae 11 0 0 0 225 Bacteria-Proteobacteria- Epsilonproteobacteria-Campylobacterales- Helicobacteraceae 15 0 0 1 31 Bacteria-Proteobacteria- Epsilonproteobacteria-Campylobacterales- Undefined 3 0 0 0 1 Bacteria-Proteobacteria- Epsilonproteobacteria-Nautiliales- Nautiliaceae 0 0 0 0 1 Bacteria-Proteobacteria- Epsilonproteobacteria-Undefined 21 0 0 0 0 Bacteria-Proteobacteria- Gammaproteobacteria-Aeromonadales- Aeromonadaceae 112 11 4 6 552 Bacteria-Proteobacteria- Gammaproteobacteria-Aeromonadales- Undefined 0 0 0 0 1 Bacteria-Proteobacteria- Gammaproteobacteria-Alteromonadales- Alteromonadaceae 10 0 0 0 3 Bacteria-Proteobacteria- Gammaproteobacteria-Alteromonadales- Idiomarinaceae 7 0 0 0 0 Bacteria-Proteobacteria- Gammaproteobacteria-Alteromonadales- Shewanellaceae 20 0 0 0 9 Bacteria-Proteobacteria- Gammaproteobacteria-Alteromonadales- Undefined 0 0 0 0 2 72

Bacteria-Proteobacteria- Gammaproteobacteria-Chromatiales- Chromatiaceae 421 28 3 3 465 Bacteria-Proteobacteria- Gammaproteobacteria-Chromatiales- Ectothiorhodospiraceae 6 0 0 0 2 Bacteria-Proteobacteria- Gammaproteobacteria-Chromatiales- Halothiobacillaceae 38 0 0 0 35 Bacteria-Proteobacteria- Gammaproteobacteria-Chromatiales- Undefined 2 0 0 0 3 Bacteria-Proteobacteria- Gammaproteobacteria-Enterobacteriales- Enterobacteriaceae 102 3444 233 8 266 Bacteria-Proteobacteria- Gammaproteobacteria- Gammaproteobacteria_incertae_sedis- Arenicella 9 0 0 0 1 Bacteria-Proteobacteria- Gammaproteobacteria- Gammaproteobacteria_incertae_sedis- Gilvimarinus 0 1 0 0 0 Bacteria-Proteobacteria- Gammaproteobacteria- Gammaproteobacteria_incertae_sedis- Sedimenticola 0 0 0 4 0 Bacteria-Proteobacteria- Gammaproteobacteria- Gammaproteobacteria_incertae_sedis- Solimonas 1 34 0 0 0 Bacteria-Proteobacteria- Gammaproteobacteria- Gammaproteobacteria_incertae_sedis- Undefined 11 1 0 52 5 Bacteria-Proteobacteria- Gammaproteobacteria-Legionellales- Coxiellaceae 141 36 33 20 20 Bacteria-Proteobacteria- Gammaproteobacteria-Legionellales- Legionellaceae 513 68 5 2 6 Bacteria-Proteobacteria- Gammaproteobacteria-Legionellales- Undefined 2 0 0 0 0 Bacteria-Proteobacteria- Gammaproteobacteria-Methylococcales- Methylococcaceae 24 6 3 5 22 Bacteria-Proteobacteria- Gammaproteobacteria-Oceanospirillales- Halomonadaceae 5 6 0 1 3 Bacteria-Proteobacteria- Gammaproteobacteria-Oceanospirillales- Oceanospirillaceae 0 4 0 0 0 73

Bacteria-Proteobacteria- Gammaproteobacteria-Oceanospirillales- Oleiphilaceae 0 0 0 0 1 Bacteria-Proteobacteria- Gammaproteobacteria-Oceanospirillales- Undefined 2 1 0 0 0 Bacteria-Proteobacteria- Gammaproteobacteria-Orbales-Orbaceae 1 0 0 0 0 Bacteria-Proteobacteria- Gammaproteobacteria-Pseudomonadales- Moraxellaceae 16 939 27 0 534 Bacteria-Proteobacteria- Gammaproteobacteria-Pseudomonadales- Pseudomonadaceae 58 610 438 17 182 Bacteria-Proteobacteria- Gammaproteobacteria-Thiotrichales- Thiotrichaceae 4 0 0 0 0 Bacteria-Proteobacteria- Gammaproteobacteria-Thiotrichales- Thiotrichales_incertae_sedis 4 1 0 0 3 Bacteria-Proteobacteria- Gammaproteobacteria-Thiotrichales- Undefined 1 0 0 0 0 Bacteria-Proteobacteria- Gammaproteobacteria-Undefined 672 100 57 69 197 Bacteria-Proteobacteria- Gammaproteobacteria-Vibrionales- Vibrionaceae 1 0 0 0 0 Bacteria-Proteobacteria- Gammaproteobacteria-Xanthomonadales- Sinobacteraceae 14 17 5 2 13 Bacteria-Proteobacteria- Gammaproteobacteria-Xanthomonadales- Undefined 3 0 0 1 1 Bacteria-Proteobacteria- Gammaproteobacteria-Xanthomonadales- Xanthomonadaceae 244 757 3532 29338 35 Bacteria-Proteobacteria-Undefined 610 134 104 195 329 Bacteria-SR1-SR1_genera_incertae_sedis- SR1_genera_incertae_sedis- SR1_genera_incertae_sedis 23 1 0 0 4 Bacteria- Saccharibacteria_genera_incertae_sedis- Saccharibacteria_genera_incertae_sedis- Saccharibacteria_genera_incertae_sedis 0 1 0 1 1 Bacteria-Spirochaetes-Spirochaetia- Spirochaetales-Brachyspiraceae 1 0 0 0 0 Bacteria-Spirochaetes-Spirochaetia- Spirochaetales-Spirochaetaceae 2 0 0 0 3 Bacteria-Spirochaetes-Spirochaetia- Spirochaetales-Spirochaetales_incertae_sedis 0 0 0 0 1 Bacteria-Synergistetes-Synergistia- Synergistales-Synergistaceae 0 0 3 0 0 74

Bacteria-Tenericutes-Mollicutes- Acholeplasmatales-Acholeplasmataceae 0 0 3 0 0 Bacteria-Undefined 1630 827 2197 5454 914 Bacteria-Verrucomicrobia-Opitutae- Opitutales-Opitutaceae 355 119 2 1 6 Bacteria-Verrucomicrobia-Opitutae- Puniceicoccales-Puniceicoccaceae 3 0 0 1 38 Bacteria-Verrucomicrobia-Opitutae- Undefined 6 1 0 0 0 Bacteria-Verrucomicrobia-Spartobacteria- Spartobacteria_genera_incertae_sedis- Spartobacteria_genera_incertae_sedis 478 11 4 70 8 Bacteria-Verrucomicrobia-Spartobacteria- Undefined 3 1 0 2 0 Bacteria-Verrucomicrobia-Spartobacteria- Xiphinematobacter-Xiphinematobacter 1 1 0 0 0 Bacteria-Verrucomicrobia-Subdivision3- Subdivision3_genera_incertae_sedis- Subdivision3_genera_incertae_sedis 30 8 1 1 11 Bacteria-Verrucomicrobia-Subdivision5- Subdivision5_genera_incertae_sedis- Subdivision5_genera_incertae_sedis 6 0 0 0 2 Bacteria-Verrucomicrobia-Undefined 24 4 0 1 2 Bacteria-Verrucomicrobia-Verrucomicrobiae- Verrucomicrobiales-Undefined 0 0 0 0 1 Bacteria-Verrucomicrobia-Verrucomicrobiae- Verrucomicrobiales-Verrucomicrobiaceae 181 9 6 0 57 Bacteria-WPS-1_genera_incertae_sedis- WPS-1_genera_incertae_sedis-WPS- 1_genera_incertae_sedis 3 3 2 1 1 Bacteria-WPS-2_genera_incertae_sedis- WPS-2_genera_incertae_sedis-WPS- 2_genera_incertae_sedis 19 11 8 1 0 Below_Threshold-Archaea 17 8 3 157 54

16S rDNA Genus Data OTUs Slag Big Creosote Big Marsh Dune Marsh RR Ditch Genus Marsh Well Well #3 SW SW Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Hydrogenophaga 326 16124 32020 33963 1131 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Undefined 39790 1119 63 0 16847 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Microbacteriaceae-Undefined 41596 514 57 0 6319 Bacteria-Firmicutes-Undefined 25 21 44830 34 11 Bacteria-Proteobacteria-Gammaproteobacteria- Xanthomonadales-Xanthomonadaceae-Silanimonas 205 93 8 28708 19 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Chitinophagaceae- Sediminibacterium 15860 135 3 5 5259 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Burkholderiaceae-Polynucleobacter 9740 5 5 0 9769 Bacteria-Proteobacteria-Alphaproteobacteria- Caulobacterales-Caulobacteraceae-Brevundimonas 70 14819 4021 2 13 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Undefined 6533 901 799 1417 8613 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Acidovorax 9404 103 50 0 4075 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Sanguibacteraceae-Sanguibacter 1 11535 1 0 0 Bacteria-Undefined 1630 827 2197 5454 914 Bacteria-Bacteroidetes-Flavobacteriia- Flavobacteriales-Flavobacteriaceae-Flavobacterium 5941 20 4 18 4009 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Intrasporangiaceae-Undefined 8299 27 11 0 1132 Bacteria-Bacteroidetes-Cytophagia-Cytophagales- Cytophagaceae-Arcicella 6789 1 1 0 2238 Bacteria-Cyanobacteria-Chloroplast-Chloroplast- Chloroplast-Chloroplast-Chlorophyta 150 29 19 7577 481 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Undefined 3878 227 335 511 2386 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Dietziaceae-Dietzia 0 6131 960 1 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodobacterales-Rhodobacteraceae-Gemmobacter 152 17 4 6200 189 Bacteria-Firmicutes-Bacilli-Bacillales- Planococcaceae-Undefined 5 5909 405 3 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Geodermatophilaceae- Blastococcus 207 5094 257 0 98 Bacteria-Firmicutes-Bacilli-Bacillales- Planococcaceae-Lysinibacillus 1 4451 143 1 0 Bacteria-Firmicutes-Bacilli-Bacillales-Bacillus 7 4232 23 4 2 Bacteria-Proteobacteria-Betaproteobacteria- Undefined 2399 104 57 66 1144 76

Bacteria-Firmicutes-Clostridia-Clostridiales- Undefined 105 22 3401 62 146 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Acetobacteraceae-Undefined 61 60 617 2833 4 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Malikia 7 464 1166 1742 164 Bacteria-Proteobacteria-Gammaproteobacteria- Xanthomonadales-Xanthomonadaceae- Stenotrophomonas 0 412 3121 3 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Geodermatophilaceae-Undefined 2123 79 7 0 1303 Bacteria-Firmicutes-Clostridia-Undefined 4 7 3310 9 4 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Micrococcaceae-Arthrobacter 1 3020 146 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Oxalobacteraceae-Massilia 1 2509 292 0 4 Bacteria-Proteobacteria-Alphaproteobacteria- Sphingomonadales-Sphingomonadaceae- Sphingomonas 37 2409 75 6 38 Bacteria-Proteobacteria-Alphaproteobacteria- Sphingomonadales-Erythrobacteraceae- Porphyrobacter 397 8 1 2061 21 Bacteria-Bacteroidetes-Flavobacteriia- Flavobacteriales-Cryomorphaceae-Undefined 1112 11 99 52 1169 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Nocardiaceae-Rhodococcus 1 1953 376 0 1 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Intrasporangiaceae-Terracoccus 2131 0 0 0 130 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Pantoea 0 2035 86 0 52 Bacteria-Firmicutes-Bacilli-Bacillales-Brevibacillus 0 2106 2 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Burkholderiales_incertae_sedis- Leptothrix 1336 2 1 0 546 Bacteria-Bacteroidetes-Undefined 556 39 132 31 1034 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Alcaligenaceae-Paenalcaligenes 0 0 1728 1 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Methylobacteriaceae-Methylobacterium 6 1541 103 0 7 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Pseudonocardiaceae- Pseudonocardia 1 1085 566 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Sphingomonadales-Sphingomonadaceae- Sphingorhabdus 188 133 0 0 1220 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Microbacteriaceae-Cryobacterium 1443 3 0 0 72 Bacteria-Proteobacteria-Betaproteobacteria- Neisseriales-Neisseriaceae-Vogesella 146 0 0 0 1341 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Micrococcaceae-Kocuria 1 1380 49 0 0 Bacteria-Firmicutes-Bacilli-Bacillales-Undefined 54 1202 139 8 4 77

Bacteria-Proteobacteria-Gammaproteobacteria- Pseudomonadales-Moraxellaceae-Acinetobacter 2 930 9 0 457 Bacteria-Proteobacteria-Undefined 610 134 104 195 329 Bacteria-Firmicutes-Bacilli-Bacillales-Paenibacillus 0 1179 80 8 1 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Nocardiaceae-Williamsia 0 1172 6 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Oxalobacteraceae-Naxibacter 0 1120 21 1 1 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Nocardioidaceae-Nocardioides 9 1043 84 0 1 Bacteria-Proteobacteria-Gammaproteobacteria- Undefined 672 100 57 69 197 Bacteria-Proteobacteria-Gammaproteobacteria- Pseudomonadales-Pseudomonadaceae-Pseudomonas 20 492 347 13 113 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Micrococcaceae-Undefined 1 935 46 1 1 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Chitinophagaceae-Undefined 558 40 4 41 293 Bacteria-Proteobacteria-Gammaproteobacteria- Chromatiales-Chromatiaceae-Rheinheimera 420 28 3 3 455 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Hyphomicrobiaceae-Devosia 9 815 58 0 4 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Burkholderiales_incertae_sedis- Undefined 505 14 14 5 322 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodobacterales-Rhodobacteraceae-Rhodobacter 327 37 0 20 457 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Burkholderiaceae-Limnobacter 733 9 2 0 92 Bacteria-Proteobacteria-Gammaproteobacteria- Xanthomonadales-Xanthomonadaceae-Undefined 16 35 149 625 9 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Tatumella 0 832 0 0 1 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Oxalobacteraceae-Undefined 273 262 12 0 284 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodobacterales-Rhodobacteraceae-Undefined 352 24 9 199 220 Bacteria-Cyanobacteria-Chloroplast-Chloroplast- Chloroplast-Chloroplast-Bacillariophyta 274 7 3 1 517 Bacteria-Proteobacteria-Betaproteobacteria- Methylophilales-Methylophilaceae-Methylophilus 129 8 0 2 645 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Undefined 79 495 44 0 142 Bacteria-Proteobacteria-Alphaproteobacteria- Undefined 460 89 46 28 98 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Burkholderiales_incertae_sedis- Aquabacterium 92 17 11 0 584 Bacteria-Bacteroidetes-Cytophagia-Cytophagales- Cytophagaceae-Undefined 465 19 0 0 203 Bacteria-Proteobacteria-Betaproteobacteria- Neisseriales-Neisseriaceae-Aquaspirillum 11 0 1 0 635 78

Bacteria-Firmicutes-Bacilli-Bacillales- Planococcaceae-Sporosarcina 0 625 19 0 0 Bacteria-Actinobacteria-Actinobacteria- Solirubrobacterales-Conexibacteraceae-Conexibacter 5 579 13 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Legionellales-Legionellaceae-Legionella 513 68 5 2 6 Bacteria-Verrucomicrobia-Spartobacteria- Spartobacteria_genera_incertae_sedis- Spartobacteria_genera_incertae_sedis- Spartobacteria_genera_incertae_sedis 478 11 4 70 8 Bacteria-Bacteroidetes-Flavobacteriia- Flavobacteriales-Flavobacteriaceae- Chryseobacterium 78 297 175 0 9 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Microbacteriaceae-Leucobacter 14 112 429 0 1 Bacteria-Proteobacteria-Betaproteobacteria- Rhodocyclales-Rhodocyclaceae-Undefined 120 226 4 48 134 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Undefined 303 111 30 1 67 Bacteria-Firmicutes-Bacilli-Bacillales- Exiguobacterium 40 441 15 0 12 Bacteria-Bacteroidetes-Cytophagia-Cytophagales- Cytophagaceae-Flectobacillus 273 0 0 0 234 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Mycobacteriaceae-Mycobacterium 153 331 13 1 5 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Microbacteriaceae-Agrococcus 447 0 0 0 31 Bacteria-Parcubacteria- Parcubacteria_genera_incertae_sedis- Parcubacteria_genera_incertae_sedis- Parcubacteria_genera_incertae_sedis- Parcubacteria_genera_incertae_sedis 304 2 0 2 154 Bacteria-Verrucomicrobia-Opitutae-Opitutales- Opitutaceae-Opitutus 341 110 2 1 6 Bacteria-Bacteroidetes-Flavobacteriia- Flavobacteriales-Cryomorphaceae-Fluviicola 148 1 2 2 297 Bacteria-Firmicutes-Clostridia-Natranaerobiales- Natranaerobiaceae-Dethiobacter 1 0 444 4 0 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Comamonas 105 11 2 0 329 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Limnohabitans 59 0 0 0 375 Bacteria-Chloroflexi-Thermomicrobia- Sphaerobacterales-Sphaerobacteraceae- Sphaerobacter 0 63 356 1 0 Bacteria-Actinobacteria-Actinobacteria- Solirubrobacterales-Patulibacteraceae-Patulibacter 0 143 273 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Acetobacteraceae-Roseomonas 32 181 6 185 7 Bacteria-Firmicutes-Bacilli-Bacillales-Cohnella 0 358 51 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Aeromonadales-Aeromonadaceae-Aeromonas 110 11 4 6 278 79

Bacteria-Firmicutes-Bacilli-Bacillales- Planococcaceae-Paenisporosarcina 0 365 25 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Rhodocyclales-Rhodocyclaceae-Shinella 8 18 331 0 1 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Micrococcaceae-Zhihengliuella 0 352 3 0 0 Bacteria-Firmicutes-Bacilli-Bacillales- Planococcaceae-Planococcaceae_incertae_sedis 0 331 23 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Acetobacteraceae-Rubritepida 0 5 71 276 0 Bacteria-Proteobacteria-Alphaproteobacteria- Sphingomonadales-Sphingomonadaceae-Undefined 88 111 7 1 145 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Oxalobacteraceae-Oxalicibacterium 48 191 1 0 112 Bacteria-Firmicutes-Bacilli-Bacillales- Staphylococcaceae-Staphylococcus 0 322 28 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Alcaligenaceae-Undefined 3 0 22 0 322 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Undefined 83 11 2 2 219 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Pseudonocardiaceae-Lentzea 0 7 307 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodobacterales-Rhodobacteraceae-Haematobacter 109 0 1 0 198 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Albidiferax 166 1 0 0 136 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Pelomonas 100 9 0 0 177 Bacteria-Firmicutes-Bacilli-Bacillales- Planococcaceae-Planomicrobium 0 273 3 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Dermabacteraceae- Brachybacterium 0 138 133 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Polaromonas 0 244 4 0 17 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Nocardioidaceae-Aeromicrobium 0 195 58 0 5 Bacteria-Cyanobacteria-Chloroplast-Chloroplast- Chloroplast-Chloroplast-Streptophyta 12 218 11 2 14 Bacteria-Proteobacteria-Gammaproteobacteria- Aeromonadales-Aeromonadaceae-Tolumonas 1 0 0 0 255 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Rhizobiaceae-Rhizobium 50 171 10 0 23 Bacteria-Proteobacteria-Alphaproteobacteria- Sphingomonadales-Sphingomonadaceae- Sphingopyxis 27 119 92 0 11 Bacteria-Bacteroidetes-Flavobacteriia- Flavobacteriales-Undefined 41 3 23 38 142 Bacteria-Actinobacteria-Actinobacteria-Undefined 62 90 19 2 72 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Saprospiraceae- Haliscomenobacter 174 0 0 0 71 80

Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Microbacteriaceae-Microcella 113 108 17 4 2 Below_Threshold-Archaea 17 8 3 157 54 Bacteria-Cyanobacteria-Chloroplast-Chloroplast- Chloroplast-Chloroplast-Undefined 24 8 0 157 39 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Hyphomicrobiaceae-Hyphomicrobium 46 44 124 2 9 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Simplicispira 150 0 0 0 75 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Sphingobacteriaceae-Pedobacter 17 155 38 0 10 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Geodermatophilaceae- Modestobacter 0 217 0 0 0 Bacteria-Cyanobacteria-Chloroplast-Cyanobacteria- Undefined 110 42 5 6 47 Bacteria-Cyanobacteria-Chloroplast-Undefined 113 1 0 64 27 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Caldimonas 0 172 9 24 0 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Burkholderiaceae-Undefined 102 1 2 0 92 Bacteria-Proteobacteria-Betaproteobacteria- Methylophilales-Methylophilaceae-Undefined 51 0 1 1 140 Bacteria-Proteobacteria-Epsilonproteobacteria- Campylobacterales-Campylobacteraceae-Arcobacter 3 0 0 0 187 Bacteria-Verrucomicrobia-Verrucomicrobiae- Verrucomicrobiales-Verrucomicrobiaceae- Luteolibacter 145 6 3 0 35 Bacteria-Proteobacteria-Gammaproteobacteria- Pseudomonadales-Pseudomonadaceae-Undefined 7 46 89 4 41 Bacteria-Bacteroidetes-Flavobacteriia- Flavobacteriales-Flavobacteriaceae-Undefined 85 1 1 0 98 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Geodermatophilaceae- Geodermatophilus 0 173 7 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Caulobacterales-Caulobacteraceae-Caulobacter 86 74 3 0 13 Bacteria-Acidobacteria-Acidobacteria_Gp4-Gp4- Gp4-Gp4 8 157 7 0 3 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Oxalobacteraceae- Noviherbaspirillum 0 158 12 0 2 Bacteria-Proteobacteria-Alphaproteobacteria- Caulobacterales-Caulobacteraceae-Asticcacaulis 97 67 0 0 6 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Microbacteriaceae- Pseudoclavibacter 14 5 2 0 145 Bacteria-Firmicutes-Clostridia-Clostridiales- Peptostreptococcaceae-Undefined 91 51 5 8 8 Bacteria-Proteobacteria-Gammaproteobacteria- Legionellales-Coxiellaceae-Coxiella 73 28 29 20 11 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Bradyrhizobiaceae-Bosea 19 51 83 0 5 81

Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Microbacteriaceae-Agromyces 0 156 1 0 0 Bacteria-Bacteroidetes-Cytophagia-Cytophagales- Undefined 58 4 0 28 64 Bacteria-Chloroflexi-Undefined 46 40 60 0 7 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Burkholderiales_incertae_sedis- Tepidimonas 4 26 55 58 9 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Brevibacteriaceae-Brevibacterium 0 45 101 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Xanthomonadales-Xanthomonadaceae-Luteimonas 2 91 51 1 1 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Micrococcaceae-Citricoccus 0 39 103 1 0 Bacteria-Bacteroidetes-Flavobacteriia- Flavobacteriales-Cryomorphaceae-Lishizhenia 40 0 0 1 102 Bacteria-Proteobacteria-Alphaproteobacteria- Sphingomonadales-Sphingomonadaceae- Novosphingobium 38 94 0 0 11 Bacteria-Proteobacteria-Gammaproteobacteria- Xanthomonadales-Xanthomonadaceae-Lysobacter 0 91 50 0 1 Bacteria-Bacteroidetes-Cytophagia-Cytophagales- Flammeovirgaceae-Cesiribacter 1 8 0 131 0 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Burkholderiales_incertae_sedis- Sphaerotilus 20 1 0 0 118 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Oxalobacteraceae-Herbaspirillum 36 15 0 0 88 Bacteria-Bacteroidetes-Cytophagia-Cytophagales- Cytophagaceae-Emticicia 18 0 0 86 33 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Oxalobacteraceae-Undibacterium 45 5 0 0 87 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Sphingobacteriaceae- Sphingobacterium 1 71 63 1 0 Bacteria-Armatimonadetes-Armatimonadia- Armatimonadales-Armatimonadaceae-Armatimonas- Armatimonadetes_gp1 37 5 0 0 88 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Variovorax 2 50 58 0 17 Bacteria-Proteobacteria-Betaproteobacteria- Gallionellales-Gallionellaceae-Sideroxydans 24 84 1 1 17 Bacteria-Actinobacteria-Actinobacteria- Solirubrobacterales-Undefined 2 101 19 1 3 Bacteria-Proteobacteria-Alphaproteobacteria- Caulobacterales-Caulobacteraceae- Phenylobacterium 4 111 0 3 4 Bacteria-Proteobacteria-Alphaproteobacteria- Sphingomonadales-Sphingomonadaceae- Sandaracinobacter 59 3 0 51 9 Bacteria-Acidobacteria-Acidobacteria_Gp6-Gp6- Gp6-Gp6 24 71 13 2 10 82

Bacteria-Actinobacteria-Actinobacteria- Solirubrobacterales-Solirubrobacteraceae- Solirubrobacter 3 93 20 3 0 Bacteria-Nitrospirae-Nitrospira-Nitrospirales- Nitrospiraceae-Nitrospira 3 114 1 0 1 Bacteria-Planctomycetes-Planctomycetia- Planctomycetales-Planctomycetaceae-Undefined 71 39 4 0 5 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Streptomycetaceae-Streptomyces 1 44 68 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Neisseriales-Neisseriaceae-Undefined 57 1 0 0 55 Bacteria-Proteobacteria-Deltaproteobacteria- Undefined 60 27 5 3 18 Bacteria-Actinobacteria-Actinobacteria- Acidimicrobiales-Undefined 8 66 27 0 10 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Kineosporiaceae- Pseudokineococcus 0 104 3 0 4 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Pseudonocardiaceae-Undefined 0 48 63 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Bogoriellaceae-Georgenia 0 81 27 0 0 Bacteria-Bacteroidetes-Cytophagia-Cytophagales- Flammeovirgaceae-Undefined 29 0 0 13 65 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Saprospiraceae-Lewinella 95 5 0 0 3 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Saprospiraceae-Undefined 80 7 0 0 15 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Burkholderiales_incertae_sedis- Inhella 79 0 0 0 23 Bacteria-Proteobacteria-Gammaproteobacteria- Xanthomonadales-Xanthomonadaceae- Pseudoxanthomonas 0 2 100 0 0 Bacteria-Actinobacteria-Actinobacteria- Acidimicrobiales-Acidimicrobineae_incertae_sedis- Aciditerrimonas 5 93 3 0 0 Bacteria-Cyanobacteria-Chloroplast-Cyanobacteria- GpXIII 2 30 2 0 67 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Cellulomonadaceae-Oerskovia 0 98 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Phyllobacteriaceae-Chelativorans 1 77 20 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Pseudomonadales-Pseudomonadaceae-Cellvibrio 26 49 0 0 22 Bacteria-Actinobacteria-Actinobacteria-Gaiellales- Gaiellaceae-Gaiella 8 77 5 6 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Beijerinckiaceae-Undefined 7 13 1 0 75 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Enterobacter 3 25 39 0 27 Bacteria-Deinococcus-Thermus-Deinococci- Deinococcales-Deinococcaceae-Deinococcus 10 73 1 0 6 83

Bacteria-Proteobacteria-Deltaproteobacteria- Desulfuromonadales-Geobacteraceae-Geobacter 37 42 0 0 11 Bacteria-Planctomycetes-Planctomycetia- Planctomycetales-Planctomycetaceae-Pirellula 48 21 1 16 3 Bacteria-Proteobacteria-Alphaproteobacteria- Sphingomonadales-Sphingomonadaceae- Sphingobium 2 83 2 0 2 Bacteria-Proteobacteria-Deltaproteobacteria- Myxococcales-Undefined 24 46 6 3 8 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Cyclobacteriaceae-Algoriphagus 28 3 1 0 53 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Rhodospirillaceae-Undefined 50 25 0 0 10 Bacteria-Proteobacteria-Alphaproteobacteria- Sphingomonadales-Erythrobacteraceae-Undefined 15 17 1 46 6 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodobacterales-Rhodobacteraceae-Paracoccus 5 66 8 0 5 Bacteria-Proteobacteria-Betaproteobacteria- Rhodocyclales-Rhodocyclaceae-Sulfuritalea 26 22 0 0 32 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Buchnera 0 11 60 8 1 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Nocardioidaceae-Undefined 1 70 7 0 0 Bacteria-Bacteroidetes-Cytophagia-Cytophagales- Cytophagaceae-Hymenobacter 1 63 7 0 6 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Bradyrhizobiaceae-Undefined 2 67 4 1 2 Bacteria-Deinococcus-Thermus-Deinococci- Deinococcales-Trueperaceae-Truepera 4 46 22 2 0 Bacteria-Acidobacteria-Acidobacteria_Gp16-Gp16- Gp16-Gp16 10 54 4 2 3 Bacteria-Acidobacteria-Acidobacteria_Gp3-Gp3- Gp3-Gp3 22 38 4 9 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Cellulomonadaceae-Cellulomonas 0 59 13 0 1 Bacteria-Proteobacteria-Gammaproteobacteria- Chromatiales-Halothiobacillaceae-Thiovirga 38 0 0 0 35 Bacteria-Proteobacteria-Alphaproteobacteria- Sphingomonadales-Sphingomonadaceae- Sphingosinicella 1 64 6 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Alcaligenaceae-Advenella 0 6 63 0 2 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Chitinophagaceae-Terrimonas 3 60 0 2 5 Bacteria-Firmicutes-Clostridia-Clostridiales- Ruminococcaceae-Undefined 16 0 12 36 6 Bacteria-Proteobacteria-Gammaproteobacteria- Gammaproteobacteria_incertae_sedis-Undefined 11 1 0 52 5 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Oxalobacteraceae- Janthinobacterium 0 68 0 0 0 Bacteria-Proteobacteria-Deltaproteobacteria- Myxococcales-Polyangiaceae-Sorangium 1 66 0 0 1 84

Bacteria-Actinobacteria-Actinobacteria- Acidimicrobiales-Iamiaceae-Iamia 0 49 18 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Nocardioidaceae-Kribbella 0 9 57 0 0 Bacteria-Firmicutes-Bacilli-Bacillales- Oceanobacillus 0 1 65 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Nocardioidaceae-Marmoricola 0 59 2 3 1 Bacteria-Cyanobacteria-Chloroplast-Chloroplast- Chloroplast-Chloroplast-Cryptomonadaceae 40 0 0 0 25 Bacteria-Proteobacteria-Betaproteobacteria- Neisseriales-Neisseriaceae-Chitinibacter 44 0 0 0 21 Bacteria-Proteobacteria-Gammaproteobacteria- Legionellales-Coxiellaceae-Aquicella 60 2 0 0 2 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Micrococcaceae-Micrococcus 0 59 3 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Promicromonosporaceae- Cellulosimicrobium 0 61 0 1 0 Bacteria-Gemmatimonadetes-Gemmatimonadetes- Gemmatimonadales-Gemmatimonadaceae- Gemmatimonas 18 37 5 0 2 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Pseudonocardiaceae-Allokutzneria 0 60 0 1 0 Bacteria-Planctomycetes-Planctomycetia- Planctomycetales-Planctomycetaceae-Aquisphaera 28 25 7 1 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Methylocystaceae-Hansschlegelia 0 0 61 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Rhodoferax 3 0 0 1 57 Bacteria-Proteobacteria-Deltaproteobacteria- Bdellovibrionales-Bdellovibrionaceae-Bdellovibrio 32 13 0 0 13 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Pseudonocardiaceae- Actinomycetospora 0 26 31 0 0 Bacteria-Fusobacteria-Fusobacteriia- Fusobacteriales-Fusobacteriaceae-Undefined 5 0 0 0 52 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodobacterales-Rhodobacteraceae- Rubellimicrobium 3 46 3 5 0 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Burkholderiaceae-Chitinimonas 21 0 0 0 36 Bacteria-Bacteroidetes- Bacteroidetes_incertae_sedis-Ohtaekwangia- Ohtaekwangia-Ohtaekwangia 47 5 0 1 3 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Roseateles 29 1 2 0 24 Bacteria-Proteobacteria-Alphaproteobacteria- Rickettsiales-Rickettsiaceae-Rickettsia 31 1 5 1 15 Bacteria-Proteobacteria-Alphaproteobacteria- Sphingomonadales-Erythrobacteraceae- Erythrobacter 52 1 0 0 0 85

Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Alcaligenaceae-Castellaniella 0 0 0 0 53 Bacteria-Proteobacteria-Gammaproteobacteria- Xanthomonadales-Xanthomonadaceae- Ignatzschineria 3 0 50 0 0 Bacteria-Cyanobacteria-Chloroplast-Cyanobacteria- GpI 0 44 7 0 1 Bacteria-Cyanobacteria-Chloroplast-Cyanobacteria- GpIX 18 11 9 0 14 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Xanthobacteraceae-Undefined 10 39 0 0 3 Bacteria-Proteobacteria-Alphaproteobacteria- Sphingomonadales-Erythrobacteraceae- Altererythrobacter 5 26 0 2 19 Archaea-Euryarchaeota-Undefined 18 1 0 0 32 Bacteria-Fusobacteria-Fusobacteriia- Fusobacteriales-Fusobacteriaceae-Propionigenium 0 0 0 0 51 Bacteria-Verrucomicrobia-Subdivision3- Subdivision3_genera_incertae_sedis- Subdivision3_genera_incertae_sedis- Subdivision3_genera_incertae_sedis 30 8 1 1 11 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Chitinophagaceae-Chitinophaga 25 7 1 0 16 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Delftia 5 22 13 0 9 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Acetobacteraceae-Saccharibacter 0 47 0 0 1 Bacteria-Proteobacteria-Betaproteobacteria- Rhodocyclales-Rhodocyclaceae-Ferribacterium 0 23 0 0 25 Bacteria-Deinococcus-Thermus-Deinococci- Thermales-Thermaceae-Meiothermus 0 6 40 1 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Lachnospiraceae-Undefined 12 11 0 16 8 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Curvibacter 21 4 0 0 22 Bacteria-Actinobacteria-Actinobacteria- Acidimicrobiales-Acidimicrobiaceae-Ilumatobacter 7 7 3 0 29 Bacteria-Chloroflexi-Anaerolineae-Anaerolineales- Anaerolineaceae-Undefined 14 10 1 1 20 Bacteria-Planctomycetes-Planctomycetia- Planctomycetales-Planctomycetaceae-Zavarzinella 24 16 0 0 5 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Methylocystaceae-Methylocystis 9 28 6 0 2 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Alcaligenaceae-Achromobacter 0 45 0 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Rhodocyclales-Rhodocyclaceae-Dechloromonas 13 0 0 0 32 Bacteria-Planctomycetes-Planctomycetia- Planctomycetales-Planctomycetaceae-Planctomyces 18 18 0 0 8 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Phyllobacteriaceae-Undefined 15 26 2 1 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Undefined 19 15 3 2 5 86

Bacteria-Proteobacteria-Gammaproteobacteria- Pseudomonadales-Moraxellaceae-Perlucidibaca 2 3 0 0 39 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Promicromonosporaceae- Promicromonospora 1 0 37 5 0 Bacteria-Bacteroidetes-Bacteroidia-Bacteroidales- Bacteroidaceae-Bacteroides 11 0 16 0 16 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Aurantimonadaceae-Aurantimonas 1 31 9 0 2 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Kineosporiaceae-Kineosporia 1 27 12 0 2 Bacteria-Proteobacteria-Betaproteobacteria- Hydrogenophilales-Hydrogenophilaceae- Thiobacillus 10 0 1 0 31 Bacteria-Firmicutes-Erysipelotrichia- Erysipelotrichales-Erysipelotrichaceae- Erysipelothrix 0 0 39 2 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Methylobacteriaceae-Microvirga 3 35 2 0 1 Bacteria-Chlamydiae-Chlamydiia-Chlamydiales- Parachlamydiaceae-Parachlamydia 32 7 0 0 1 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Hyphomicrobiaceae-Undefined 29 6 1 0 4 Bacteria-Proteobacteria-Deltaproteobacteria- Myxococcales-Polyangiaceae-Undefined 6 20 9 1 4 Bacteria-Proteobacteria-Epsilonproteobacteria- Campylobacterales-Campylobacteraceae- Sulfurospirillum 8 0 0 0 32 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Bradyrhizobiaceae-Bradyrhizobium 6 24 5 2 2 Bacteria-WPS-2_genera_incertae_sedis-WPS- 2_genera_incertae_sedis-WPS- 2_genera_incertae_sedis-WPS- 2_genera_incertae_sedis 19 11 8 1 0 Bacteria-Chlamydiae-Chlamydiia-Chlamydiales- Undefined 19 13 0 3 3 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Bradyrhizobiaceae-Rhodopseudomonas 0 32 4 1 1 Bacteria-Chlamydiae-Chlamydiia-Chlamydiales- Simkaniaceae-Simkania 26 7 0 2 2 Bacteria-Proteobacteria-Alphaproteobacteria- Caulobacterales-Caulobacteraceae-Undefined 8 19 8 0 2 Bacteria-Firmicutes-Bacilli-Lactobacillales- Carnobacteriaceae-Carnobacterium 7 1 0 28 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Beijerinckiaceae-Beijerinckia 2 28 1 4 1 Bacteria-Proteobacteria-Gammaproteobacteria- Methylococcales-Methylococcaceae-Undefined 13 6 3 1 13 Bacteria-Proteobacteria-Gammaproteobacteria- Xanthomonadales-Sinobacteraceae-Steroidobacter 11 7 5 2 11 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Brucellaceae-Ochrobactrum 0 20 15 0 0 87

Bacteria-Proteobacteria-Gammaproteobacteria- Gammaproteobacteria_incertae_sedis-Solimonas- Solimonas 1 34 0 0 0 Bacteria-Verrucomicrobia-Verrucomicrobiae- Verrucomicrobiales-Verrucomicrobiaceae- Undefined 23 0 3 0 9 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Microbacteriaceae-Curtobacterium 1 21 9 0 3 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Nakamurellaceae-Nakamurella 0 26 7 0 1 Bacteria-Proteobacteria-Alphaproteobacteria- Sphingomonadales-Undefined 10 5 2 11 6 Bacteria-Cyanobacteria-Chloroplast-Cyanobacteria- GpVI 29 0 0 0 4 Bacteria-Proteobacteria-Alphaproteobacteria- Caulobacterales-Hyphomonadaceae-Hyphomonas 26 2 0 0 5 Bacteria-Proteobacteria-Epsilonproteobacteria- Campylobacterales-Helicobacteraceae- Sulfuricurvum 11 0 0 0 22 Bacteria-Proteobacteria-Gammaproteobacteria- Pseudomonadales-Moraxellaceae-Undefined 2 0 0 0 31 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Caenimonas 14 0 0 0 18 Bacteria-Proteobacteria-Deltaproteobacteria- Bdellovibrionales-Bacteriovoracaceae-Peredibacter 20 2 6 0 4 Bacteria-Proteobacteria-Deltaproteobacteria- Bdellovibrionales-Bdellovibrionaceae- Vampirovibrio 14 10 0 3 5 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Cellulomonadaceae-Undefined 0 30 1 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Corynebacteriaceae- Corynebacterium 1 16 13 0 1 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae- Pseudorhodoferax 1 23 6 1 0 Bacteria-Verrucomicrobia-Undefined 24 4 0 1 2 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Micrococcaceae-Acaricomes 0 30 0 0 0 Bacteria-Firmicutes-Clostridia-Clostridiales-Sarcina 27 0 0 2 1 Bacteria-Proteobacteria-Betaproteobacteria- Rhodocyclales-Rhodocyclaceae-Azonexus 6 0 0 0 24 Bacteria-Proteobacteria-Deltaproteobacteria- Myxococcales-Cystobacteraceae-Anaeromyxobacter 10 17 0 0 3 Archaea-Thaumarchaeota-Nitrososphaerales- Nitrososphaerales-Nitrososphaeraceae- Nitrososphaera 3 24 0 2 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Microbacteriaceae-Leifsonia 0 29 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Rubrobacterales-Rubrobacteraceae-Rubrobacter 0 26 3 0 0 Bacteria-Chloroflexi-Chloroflexia-Chloroflexales- Oscillochloridaceae-Oscillochloris 24 5 0 0 0 88

Bacteria-Proteobacteria-Gammaproteobacteria- Alteromonadales-Shewanellaceae-Shewanella 20 0 0 0 9 Bacteria-Chloroflexi-Thermomicrobia-Undefined 0 8 20 0 0 Bacteria-Firmicutes-Bacilli-Bacillales- Planococcaceae-Chryseomicrobium 1 25 2 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Acetobacteraceae-Belnapia 10 16 0 1 1 Bacteria-Proteobacteria-Alphaproteobacteria- Rickettsiales-Rickettsiaceae-Orientia 4 0 0 0 24 Bacteria-Proteobacteria-Alphaproteobacteria- SAR11-SAR11-Undefined 0 0 0 0 28 Bacteria-SR1-SR1_genera_incertae_sedis- SR1_genera_incertae_sedis- SR1_genera_incertae_sedis- SR1_genera_incertae_sedis 23 1 0 0 4 Bacteria-Bacteroidetes-Cytophagia-Cytophagales- Cytophagaceae-Dyadobacter 0 26 1 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Phyllobacteriaceae-Aquamicrobium 0 0 27 0 0 Bacteria-Verrucomicrobia-Opitutae-Puniceicoccales- Puniceicoccaceae-Cerasicoccus 0 0 0 0 27 Bacteria-Acidobacteria-Acidobacteria_Gp4- Blastocatella-Blastocatella-Blastocatella 5 8 12 0 1 Bacteria-Acidobacteria-Acidobacteria_Gp7-Gp7- Gp7-Gp7 3 19 3 1 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Kineosporiaceae-Kineococcus 0 24 1 0 1 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Microbacteriaceae-Herbiconiux 3 3 20 0 0 Bacteria-Bacteroidetes-Cytophagia-Cytophagales- Cytophagaceae-Adhaeribacter 1 24 1 0 0 Bacteria-Bacteroidetes-Flavobacteriia- Flavobacteriales-Cryomorphaceae-Wandonia 9 0 4 3 10 Bacteria-Bacteroidetes-Flavobacteriia- Flavobacteriales-Flavobacteriaceae-Cloacibacterium 12 4 0 0 10 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Methylocystaceae-Methylopila 0 16 10 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Xanthobacteraceae-Pseudolabrys 1 17 5 3 0 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Alcaligenaceae-Azohydromonas 12 2 1 0 11 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Alcaligenaceae-Pigmentiphaga 1 21 4 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Rhodocyclales-Rhodocyclaceae-Uliginosibacterium 18 0 0 0 8 Bacteria-Bacteroidetes-Cytophagia-Cytophagales- Flammeovirgaceae-Marinoscillum 0 0 0 0 25 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Chitinophagaceae- Ferruginibacter 13 4 1 1 6 Bacteria-Firmicutes-Bacilli-Bacillales- Planococcaceae-Planococcus 0 21 4 0 0 89

Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Hyphomicrobiaceae-Aquabacter 23 2 0 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Nitrosomonadales-Nitrosomonadaceae-Nitrosospira 6 17 0 0 2 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Intrasporangiaceae-Janibacter 0 17 6 0 1 Bacteria-Bacteroidetes-Bacteroidia-Bacteroidales- Porphyromonadaceae-Parabacteroides 5 5 11 2 1 Bacteria-Bacteroidetes-Bacteroidia-Bacteroidales- Undefined 12 1 3 1 7 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Cyclobacteriaceae-Aquiflexum 2 10 0 12 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Rhizobiaceae-Ensifer 1 21 0 0 2 Bacteria-Proteobacteria-Gammaproteobacteria- Pseudomonadales-Pseudomonadaceae-Rhizobacter 4 18 0 0 2 Bacteria-Firmicutes-Bacilli-Lactobacillales- Streptococcaceae-Lactococcus 2 6 0 1 14 Bacteria-Proteobacteria-Deltaproteobacteria- Syntrophobacterales-Syntrophaceae-Desulfomonile 8 12 0 0 3 Bacteria-Proteobacteria-Gammaproteobacteria- Legionellales-Coxiellaceae-Diplorickettsia 6 6 4 0 7 Bacteria-Bacteroidetes-Bacteroidia-Bacteroidales- Porphyromonadaceae-Paludibacter 11 0 0 0 11 Bacteria-Firmicutes-Clostridia-Clostridiales- Tissierella 0 0 22 0 0 Bacteria-Fusobacteria-Fusobacteriia- Fusobacteriales-Fusobacteriaceae-Cetobacterium 18 0 0 0 4 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Hyphomicrobiaceae-Filomicrobium 22 0 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodobacterales-Rhodobacteraceae-Rubribacterium 18 0 0 0 4 Bacteria-Proteobacteria-Betaproteobacteria- Neisseriales-Neisseriaceae-Deefgea 13 0 0 0 9 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Demequinaceae-Demequina 10 11 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Nocardiopsaceae-Nocardiopsis 0 12 9 0 0 Bacteria-Actinobacteria-Actinobacteria- Coriobacteriales-Coriobacteriaceae-Olsenella 0 19 1 0 1 Bacteria-Armatimonadetes-Armatimonadetes_gp5- Armatimonadetes_gp5-Armatimonadetes_gp5- Armatimonadetes_gp5 6 9 0 5 1 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Chitinophagaceae-Lacibacter 2 19 0 0 0 Bacteria-Firmicutes-Bacilli-Undefined 1 16 3 1 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Methylocystaceae-Undefined 0 2 18 0 1 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Phyllobacteriaceae-Mesorhizobium 0 18 0 0 3 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Rhodospirillaceae-Skermanella 0 21 0 0 0 90

Bacteria-Proteobacteria-Epsilonproteobacteria- Undefined 21 0 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Micromonosporaceae-Undefined 2 12 4 0 2 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Streptomycetaceae-Undefined 0 9 10 1 0 Bacteria-Actinobacteria-Undefined 1 17 1 0 1 Bacteria-Bacteroidetes-Cytophagia-Cytophagales- Cytophagaceae-Runella 9 0 0 0 11 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Beijerinckiaceae-Methylorosula 3 0 0 0 17 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfobacterales-Desulfobulbaceae-Desulfocapsa 12 0 0 0 8 Bacteria-Proteobacteria-Gammaproteobacteria- Aeromonadales-Aeromonadaceae-Undefined 1 0 0 0 19 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Citrobacter 0 17 1 0 2 Bacteria-Acidobacteria-Acidobacteria_Gp1-Gp1- Gp1-Gp1 6 0 2 8 3 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Intrasporangiaceae-Terrabacter 17 0 0 0 2 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Nocardiaceae-Nocardia 0 19 0 0 0 Bacteria-Planctomycetes-Planctomycetia- Planctomycetales-Planctomycetaceae-Rhodopirellula 12 1 0 5 1 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Bradyrhizobiaceae-Afipia 0 18 0 0 1 Bacteria-Proteobacteria-Gammaproteobacteria- Pseudomonadales-Moraxellaceae-Alkanindiges 10 5 0 0 4 Bacteria-Proteobacteria-Gammaproteobacteria- Xanthomonadales-Xanthomonadaceae- Panacagrimonas 0 18 0 0 1 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Promicromonosporaceae- Undefined 3 5 10 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Pseudonocardiaceae- Amycolatopsis 0 1 17 0 0 Bacteria-Bacteroidetes-Bacteroidia-Bacteroidales- Porphyromonadaceae-Undefined 6 0 0 0 12 Bacteria-Firmicutes-Bacilli-Bacillales- Anaerobacillus 0 18 0 0 0 Bacteria-Firmicutes-Bacilli-Bacillales- Paraliobacillus 0 16 2 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodobacterales-Rhodobacteraceae-Amaricoccus 13 1 0 0 4 Bacteria-Proteobacteria-Gammaproteobacteria- Pseudomonadales-Moraxellaceae-Psychrobacter 0 0 18 0 0 Archaea-Undefined 7 0 0 0 10 Bacteria-Actinobacteria-Thermoleophilia- Thermoleophilales-Thermoleophilaceae- Thermoleophilum 0 15 0 1 1 91

Bacteria-Planctomycetes-Planctomycetia- Planctomycetales-Planctomycetaceae-Gemmata 7 8 1 0 1 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Oxalobacteraceae-Duganella 0 15 2 0 0 Bacteria-Proteobacteria-Deltaproteobacteria- Myxococcales-Polyangiaceae-Chondromyces 6 10 1 0 0 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Chitinophagaceae- Flavisolibacter 1 13 0 1 1 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Acetobacteraceae-Paracraurococcus 5 2 0 9 0 Bacteria-Proteobacteria-Betaproteobacteria- Rhodocyclales-Rhodocyclaceae-Azospira 7 0 0 0 9 Bacteria-Proteobacteria-Deltaproteobacteria- Bdellovibrionales-Bacteriovoracaceae-Bacteriovorax 7 1 0 0 8 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Micrococcaceae-Renibacterium 0 10 5 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Streptomycetaceae- Streptacidiphilus 0 1 14 0 0 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Chitinophagaceae- Parasegetibacter 0 13 0 1 1 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Sphingobacteriaceae-Solitalea 13 0 0 0 2 Bacteria-Firmicutes-Bacilli-Bacillales- Planococcaceae-Psychrobacillus 0 2 13 0 0 Bacteria-Firmicutes-Bacilli-Lactobacillales- Carnobacteriaceae-Dolosigranulum 0 15 0 0 0 Bacteria-Firmicutes-Negativicutes- Selenomonadales-Veillonellaceae-Undefined 6 0 2 3 4 Bacteria-Microgenomates- Microgenomates_genera_incertae_sedis- Microgenomates_genera_incertae_sedis- Microgenomates_genera_incertae_sedis- Microgenomates_genera_incertae_sedis 1 0 0 0 14 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Acetobacteraceae-Acidisoma 2 10 2 0 1 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Rhodospirillaceae-Azospirillum 5 2 0 0 8 Bacteria-Proteobacteria-Betaproteobacteria- Rhodocyclales-Rhodocyclaceae-Thauera 4 4 0 0 7 Bacteria-Proteobacteria-Gammaproteobacteria- Xanthomonadales-Xanthomonadaceae-Aquimonas 12 0 0 0 3 Bacteria-Verrucomicrobia-Opitutae-Puniceicoccales- Puniceicoccaceae-Undefined 3 0 0 1 11 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Microbacteriaceae-Frondihabitans 0 9 5 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Nocardiaceae-Undefined 0 11 3 0 0 Bacteria-Actinobacteria-Actinobacteria- Coriobacteriales-Coriobacteriaceae-Undefined 6 3 4 0 1 92

Bacteria-Bacteroidetes-Flavobacteriia- Flavobacteriales-Flavobacteriaceae-Moheibacter 0 0 14 0 0 Bacteria-Chlamydiae-Chlamydiia-Chlamydiales- Parachlamydiaceae-Undefined 10 4 0 0 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Ruminococcaceae-Saccharofermentans 6 0 4 0 4 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Hyphomicrobiaceae-Blastochloris 2 12 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Sphingomonadales-Erythrobacteraceae- Erythromicrobium 4 0 0 10 0 Bacteria-Proteobacteria-Alphaproteobacteria- Sphingomonadales-Sphingomonadaceae- Sandarakinorhabdus 4 0 0 0 10 Bacteria-Proteobacteria-Betaproteobacteria- Neisseriales-Neisseriaceae-Andreprevotia 10 0 0 0 4 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfobacterales-Desulfobacteraceae-Undefined 8 0 0 0 6 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Yersinia 0 0 0 0 14 Bacteria-Proteobacteria-Gammaproteobacteria- Oceanospirillales-Halomonadaceae-Halomonas 4 6 0 1 3 Bacteria-Verrucomicrobia-Opitutae-Opitutales- Opitutaceae-Undefined 8 6 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Kineosporiaceae-Undefined 0 12 1 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Micromonosporaceae- Micromonospora 0 10 3 0 0 Bacteria-Bacteroidetes-Cytophagia-Cytophagales- Cytophagaceae-Spirosoma 1 5 2 0 5 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Sphingobacteriaceae- Mucilaginibacter 1 3 0 0 9 Bacteria-Proteobacteria-Alphaproteobacteria- Alphaproteobacteria_incertae_sedis-Geminicoccus- Geminicoccus 4 7 1 0 1 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Phyllobacteriaceae-Aminobacter 7 5 0 0 1 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Oxalobacteraceae-Herminiimonas 8 3 0 1 1 Bacteria-Proteobacteria-Betaproteobacteria- Rhodocyclales-Rhodocyclaceae-Propionivibrio 1 4 0 0 8 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Arsenophonus 12 0 0 0 1 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Salmonella 0 3 0 0 10 Bacteria-Verrucomicrobia-Verrucomicrobiae- Verrucomicrobiales-Verrucomicrobiaceae- Prosthecobacter 4 3 0 0 6 Bacteria-Acidobacteria-Acidobacteria_Gp17-Gp17- Gp17-Gp17 1 7 1 0 3 93

Bacteria-Acidobacteria-Acidobacteria_Gp4- Undefined 0 12 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Dermabacteraceae-Undefined 0 5 7 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Promicromonosporaceae- Isoptericola 0 10 2 0 0 Bacteria-Bacteroidetes-Flavobacteriia- Flavobacteriales-Flavobacteriaceae-Epilithonimonas 1 4 5 0 2 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Sphingobacteriaceae- Arcticibacter 0 11 0 0 1 Bacteria-Chlamydiae-Chlamydiia-Chlamydiales- Parachlamydiaceae-Neochlamydia 7 3 0 2 0 Bacteria-Planctomycetes-Planctomycetia- Planctomycetales-Planctomycetaceae-Blastopirellula 11 0 0 0 1 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Beijerinckiaceae-Chelatococcus 0 12 0 0 0 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfovibrionales-Desulfovibrionaceae- Desulfovibrio 7 0 1 0 4 Bacteria-Proteobacteria-Gammaproteobacteria- Xanthomonadales-Xanthomonadaceae-Arenimonas 6 6 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Acidimicrobiales-Acidimicrobiaceae-Undefined 0 5 5 0 1 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Cryptosporangiaceae- Cryptosporangium 0 11 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Intrasporangiaceae- Ornithinimicrobium 0 1 9 0 1 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Micrococcaceae-Rothia 0 9 0 2 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Ruaniaceae-Haloactinobacterium 0 9 2 0 0 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Chitinophagaceae-Flavitalea 2 9 0 0 0 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Chitinophagaceae-Hydrotalea 9 2 0 0 0 Bacteria-Chloroflexi-Anaerolineae-Anaerolineales- Anaerolineaceae-Longilinea 5 1 1 0 4 Bacteria-Firmicutes-Clostridia-Clostridiales- Desulfosporosinus 2 2 4 3 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Methylobacteriaceae-Undefined 1 9 1 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Burkholderiales_incertae_sedis- Ideonella 1 0 0 0 10 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Ramlibacter 1 10 0 0 0 Bacteria-Acidobacteria-Acidobacteria_Gp2-Gp2- Gp2-Gp2 4 0 0 5 1 94

Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Demequinaceae-Undefined 7 0 0 0 3 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Intrasporangiaceae-Phycicoccus 0 10 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Microbacteriaceae-Clavibacter 0 9 0 0 1 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Micrococcaceae-Nesterenkonia 0 9 1 0 0 Bacteria-Bacteroidetes-Bacteroidia-Bacteroidales- Porphyromonadaceae-Dysgonomonas 9 0 0 0 1 Bacteria-Bacteroidetes-Cytophagia-Cytophagales- Cytophagaceae-Cytophaga 2 7 0 0 1 Bacteria-Chloroflexi-Caldilineae-Caldilineales- Caldilineaceae-Litorilinea 4 2 1 0 3 Bacteria-Firmicutes-Bacilli-Bacillales- Planococcaceae-Viridibacillus 0 7 1 0 2 Bacteria-Firmicutes-Clostridia-Clostridiales- Lachnospiraceae-Cellulosilyticum 9 0 0 0 1 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Hyphomicrobiaceae-Rhodoplanes 6 2 1 0 1 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Phyllobacteriaceae-Hoeflea 0 9 1 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Rhodospirillaceae- Magnetospirillum 4 0 0 0 6 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Burkholderiales_incertae_sedis- Aquincola 1 0 0 0 9 Bacteria-Proteobacteria-Deltaproteobacteria- Syntrophobacterales-Syntrophobacteraceae- Desulfacinum 0 10 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Gammaproteobacteria_incertae_sedis-Arenicella- Arenicella 9 0 0 0 1 Bacteria-Proteobacteria-Gammaproteobacteria- Methylococcales-Methylococcaceae-Methylobacter 7 0 0 0 3 Bacteria-WPS-1_genera_incertae_sedis-WPS- 1_genera_incertae_sedis-WPS- 1_genera_incertae_sedis-WPS- 1_genera_incertae_sedis 3 3 2 1 1 Bacteria-Acidobacteria-Acidobacteria_Gp10-Gp10- Gp10-Gp10 5 2 1 0 1 Bacteria-Cyanobacteria-Chloroplast-Cyanobacteria- GpIV 8 0 1 0 0 Bacteria-Cyanobacteria-Chloroplast-Cyanobacteria- GpV 9 0 0 0 0 Bacteria-Cyanobacteria-Chloroplast-Cyanobacteria- GpXI 9 0 0 0 0 Bacteria-Firmicutes-Bacilli-Lactobacillales- Carnobacteriaceae-Atopostipes 0 0 9 0 0 Bacteria-Ignavibacteriae-Ignavibacteria- Ignavibacteriales-Ignavibacteriaceae- Ignavibacterium 3 3 0 0 3 95

Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Acetobacteraceae-Roseococcus 0 0 0 9 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rickettsiales-Anaplasmataceae-Neorickettsia 9 0 0 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Methylophilales-Methylophilaceae-Methylobacillus 5 3 0 0 1 Bacteria-Proteobacteria-Betaproteobacteria- Rhodocyclales-Rhodocyclaceae-Zoogloea 0 5 0 0 4 Bacteria-Proteobacteria-Deltaproteobacteria- Myxococcales-Myxococcaceae-Corallococcus 0 5 3 1 0 Bacteria-Proteobacteria-Deltaproteobacteria- Myxococcales-Polyangiaceae-Byssovorax 1 6 1 1 0 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Escherichia- Shigella 0 8 1 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Methylococcales-Methylococcaceae-Methylomonas 2 0 0 4 3 Bacteria-Verrucomicrobia-Opitutae-Opitutales- Opitutaceae-Alterococcus 6 3 0 0 0 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Chitinophagaceae-Segetibacter 0 7 1 0 0 Bacteria-Firmicutes-Bacilli-Bacillales- Ammoniphilus 0 8 0 0 0 Bacteria-Firmicutes-Bacilli-Lactobacillales- Carnobacteriaceae-Lacticigenium 0 0 0 8 0 Bacteria-Firmicutes-Bacilli-Lactobacillales- Enterococcaceae-Undefined 0 0 0 8 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Natranaerovirga-Natranaerovirga 0 0 0 0 8 Bacteria-Proteobacteria-Alphaproteobacteria- Rickettsiales-Rickettsiaceae-Undefined 6 1 0 0 1 Bacteria-Proteobacteria-Betaproteobacteria- Neisseriales-Neisseriaceae-Aquitalea 8 0 0 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Neisseriales-Neisseriaceae-Chromobacterium 1 0 0 0 7 Bacteria-Proteobacteria-Gammaproteobacteria- Alteromonadales-Alteromonadaceae-Haliea 5 0 0 0 3 Bacteria-Proteobacteria-Gammaproteobacteria- Chromatiales-Ectothiorhodospiraceae-Undefined 6 0 0 0 2 Bacteria-Proteobacteria-Gammaproteobacteria- Xanthomonadales-Xanthomonadaceae- Xanthomonas 0 7 1 0 0 Bacteria-Verrucomicrobia-Subdivision5- Subdivision5_genera_incertae_sedis- Subdivision5_genera_incertae_sedis- Subdivision5_genera_incertae_sedis 6 0 0 0 2 Bacteria-Verrucomicrobia-Verrucomicrobiae- Verrucomicrobiales-Verrucomicrobiaceae- Haloferula 6 0 0 0 2 Bacteria-Acidobacteria-Acidobacteria_Gp1- Undefined 3 2 1 0 1 Bacteria-Acidobacteria-Acidobacteria_Gp3- Undefined 3 2 0 1 1 96

Bacteria-Acidobacteria-Holophagae-Holophagales- Holophagaceae-Geothrix 3 0 0 0 4 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Microbacteriaceae-Alpinimonas 5 0 0 0 2 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Microbacteriaceae-Microbacterium 0 3 4 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Micrococcaceae-Sinomonas 0 2 5 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Micromonosporaceae- Actinoplanes 2 1 4 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Nocardiaceae-Gordonia 0 7 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Nocardioidaceae-Pimelobacter 0 5 2 0 0 Bacteria-Bacteroidetes-Flavobacteriia- Flavobacteriales-Cryomorphaceae-Brumimicrobium 3 0 0 0 4 Bacteria-Chloroflexi-Ktedonobacteria- Ktedonobacterales-Ktedonobacteraceae- Ktedonobacter 1 1 1 3 1 Bacteria-Firmicutes-Bacilli-Lactobacillales- Streptococcaceae-Streptococcus 0 6 1 0 0 Bacteria-Fusobacteria-Fusobacteriia- Fusobacteriales-Fusobacteriaceae-Fusobacterium 1 3 0 0 3 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Methylobacteriaceae-Meganema 7 0 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Rhodospirillaceae-Rhodocista 3 0 0 0 4 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Diaphorobacter 2 0 0 1 4 Bacteria-Proteobacteria-Epsilonproteobacteria- Campylobacterales-Helicobacteraceae-Sulfurimonas 0 0 0 0 7 Bacteria-Proteobacteria-Gammaproteobacteria- Alteromonadales-Idiomarinaceae-Pseudidiomarina 7 0 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Dickeya 2 0 0 0 5 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Erwinia 0 7 0 0 0 Bacteria-Verrucomicrobia-Opitutae-Undefined 6 1 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Kineosporiaceae-Quadrisphaera 0 6 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Mycobacteriaceae-Undefined 5 1 0 0 0 Bacteria-Chloroflexi-Chloroflexia-Chloroflexales- Undefined 1 5 0 0 0 Bacteria-Firmicutes-Bacilli-Lactobacillales- Enterococcaceae-Enterococcus 0 2 2 2 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Peptostreptococcaceae-Acetoanaerobium 0 1 1 4 0 Bacteria-Firmicutes-Erysipelotrichia- Erysipelotrichales-Erysipelotrichaceae-Undefined 2 3 1 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Aurantimonadaceae-Undefined 1 5 0 0 0 97

Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Bradyrhizobiaceae-Nitrobacter 2 3 1 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Rhodobiaceae-Rhodoligotrophos 5 0 0 0 1 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodobacterales-Rhodobacteraceae-Labrenzia 6 0 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodobacterales-Rhodobacteraceae-Rubrimonas 6 0 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Rhodospirillaceae-Novispirillum 2 0 0 0 4 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Burkholderiaceae-Cupriavidus 0 4 2 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Sutterellaceae-Sutterella 0 0 0 0 6 Bacteria-Proteobacteria-Betaproteobacteria- Neisseriales-Neisseriaceae-Pseudogulbenkiania 0 0 0 0 6 Bacteria-Proteobacteria-Betaproteobacteria- Rhodocyclales-Rhodocyclaceae-Azoarcus 3 1 0 0 2 Bacteria-Proteobacteria-Epsilonproteobacteria- Campylobacterales-Campylobacteraceae-Undefined 0 0 0 0 6 Bacteria-Proteobacteria-Gammaproteobacteria- Chromatiales-Chromatiaceae-Marichromatium 1 0 0 0 5 Bacteria-Proteobacteria-Gammaproteobacteria- Pseudomonadales-Pseudomonadaceae-Serpens 0 5 1 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Xanthomonadales-Sinobacteraceae- Hydrocarboniphaga 0 6 0 0 0 Bacteria-Verrucomicrobia-Spartobacteria-Undefined 3 1 0 2 0 Bacteria-Verrucomicrobia-Verrucomicrobiae- Verrucomicrobiales-Verrucomicrobiaceae- Akkermansia 1 0 0 0 5 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Microbacteriaceae-Rathayibacter 2 3 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Micromonosporaceae- Couchioplanes 0 4 1 0 0 Bacteria-Armatimonadetes-Armatimonadetes_gp2- Armatimonadetes_gp2-Armatimonadetes_gp2- Armatimonadetes_gp2 3 0 2 0 0 Bacteria-Bacteroidetes-Bacteroidia-Bacteroidales- Bacteroidaceae-Anaerorhabdus 0 0 4 1 0 Bacteria-Bacteroidetes-Flavobacteriia- Flavobacteriales-Flavobacteriaceae- Ornithobacterium 5 0 0 0 0 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Chitinophagaceae- Flavihumibacter 3 1 0 0 1 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Cyclobacteriaceae-Undefined 0 1 0 3 1 Bacteria-Firmicutes-Bacilli-Bacillales- Alicyclobacillaceae-Tumebacillus 0 3 2 0 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Anaerobacter 1 0 1 0 3 98

Bacteria-Firmicutes-Clostridia-Clostridiales- Peptostreptococcaceae-Peptostreptococcus 0 0 5 0 0 Bacteria-Firmicutes-Erysipelotrichia- Erysipelotrichales-Erysipelotrichaceae-Allobaculum 0 0 0 0 5 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Rhizobiales_incertae_sedis-Bauldia 0 5 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Rhizobiales_incertae_sedis-Undefined 3 0 0 0 2 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Rhizobiales_incertae_sedis-Vasilyevaea 1 2 2 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodobacterales-Rhodobacteraceae-Rhodovulum 5 0 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Acetobacteraceae-Acidicaldus 0 4 1 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Acetobacteraceae-Craurococcus 0 5 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Rhodospirillaceae-Pelagibius 4 1 0 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Kinneretia 1 0 0 0 4 Bacteria-Proteobacteria-Betaproteobacteria- Methylophilales-Methylophilaceae-Methylotenera 1 0 0 1 3 Bacteria-Proteobacteria-Deltaproteobacteria- Syntrophobacterales-Syntrophaceae-Undefined 4 0 0 0 1 Bacteria-Proteobacteria-Deltaproteobacteria- Syntrophobacterales-Syntrophobacteraceae- Desulfovirga 5 0 0 0 0 Bacteria-Proteobacteria-Epsilonproteobacteria- Campylobacterales-Helicobacteraceae-Undefined 4 0 0 0 1 Bacteria-Proteobacteria-Gammaproteobacteria- Chromatiales-Undefined 2 0 0 0 3 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Plesiomonas 3 0 0 0 2 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Pluralibacter 0 4 0 0 1 Bacteria-Proteobacteria-Gammaproteobacteria- Xanthomonadales-Sinobacteraceae-Nevskia 3 0 0 0 2 Bacteria-Proteobacteria-Gammaproteobacteria- Xanthomonadales-Undefined 3 0 0 1 1 Bacteria-Acidobacteria-Acidobacteria_Gp3- Bryobacter-Bryobacter-Bryobacter 4 0 0 0 0 Bacteria-Acidobacteria-Undefined 2 2 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Intrasporangiaceae- Intrasporangium 0 4 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Microbacteriaceae-Chryseoglobus 3 0 1 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Sporichthyaceae-Sporichthya 4 0 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Thermomonosporaceae- Actinomadura 0 4 0 0 0 99

Bacteria-Actinobacteria-Actinobacteria- Nitriliruptorales-Nitriliruptoraceae-Nitriliruptor 0 4 0 0 0 Bacteria-Bacteroidetes-Bacteroidia-Bacteroidales- Prevotellaceae-Prevotella 0 0 0 0 4 Bacteria-Bacteroidetes-Bacteroidia-Bacteroidales- Sunxiuqinia-Sunxiuqinia 4 0 0 0 0 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Rhodothermaceae-Salinibacter 1 1 2 0 0 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Sphingobacteriaceae-Undefined 3 0 0 0 1 Bacteria-Chloroflexi-Anaerolineae-Anaerolineales- Anaerolineaceae-Bellilinea 0 0 0 0 4 Bacteria-Chloroflexi-Anaerolineae-Anaerolineales- Anaerolineaceae-Leptolinea 2 0 0 0 2 Bacteria-Firmicutes-Bacilli-Bacillales- Halolactibacillus 0 4 0 0 0 Bacteria-Firmicutes-Bacilli-Bacillales- Planococcaceae-Caryophanon 0 0 4 0 0 Bacteria-Firmicutes-Bacilli-Bacillales- Saccharibacillus 1 0 0 1 2 Bacteria-Firmicutes-Bacilli-Lactobacillales- Carnobacteriaceae-Undefined 0 0 0 4 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Anaerococcus 0 4 0 0 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Anaerovorax 0 1 0 2 1 Bacteria-Firmicutes-Clostridia-Clostridiales- Peptostreptococcaceae-Tepidibacter 4 0 0 0 0 Bacteria-Lentisphaerae-Lentisphaeria-Victivallales- Victivallaceae-Victivallis 2 0 0 0 2 Bacteria-Planctomycetes-Planctomycetia-Undefined 2 1 0 0 1 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Hyphomicrobiaceae-Pedomicrobium 3 0 1 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodobacterales-Rhodobacteraceae-Salipiger 2 0 0 0 2 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Acetobacteraceae-Humitalea 0 0 4 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rickettsiales-Undefined 3 0 0 0 1 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Alcaligenaceae-Bordetella 1 3 0 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Neisseriales-Neisseriaceae-Gulbenkiania 1 0 0 0 3 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfobacterales-Desulfobacteraceae-Desulforegula 4 0 0 0 0 Bacteria-Proteobacteria-Deltaproteobacteria- Myxococcales-Cystobacteraceae-Undefined 0 3 1 0 0 Bacteria-Proteobacteria-Deltaproteobacteria- Syntrophobacterales-Syntrophaceae-Smithella 3 0 0 1 0 Bacteria-Proteobacteria-Epsilonproteobacteria- Campylobacterales-Undefined 3 0 0 0 1 100

Bacteria-Proteobacteria-Gammaproteobacteria- Alteromonadales-Alteromonadaceae- Saccharophagus 4 0 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Cronobacter 0 3 0 0 1 Bacteria-Proteobacteria-Gammaproteobacteria- Gammaproteobacteria_incertae_sedis- Sedimenticola-Sedimenticola 0 0 0 4 0 Bacteria-Proteobacteria-Gammaproteobacteria- Oceanospirillales-Oceanospirillaceae-Marinomonas 0 4 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Pseudomonadales-Moraxellaceae-Paraperlucidibaca 0 1 0 0 3 Bacteria-Proteobacteria-Gammaproteobacteria- Thiotrichales-Thiotrichaceae-Thiothrix 4 0 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Thiotrichales-Thiotrichales_incertae_sedis- Caedibacter 0 1 0 0 3 Bacteria-Proteobacteria-Gammaproteobacteria- Thiotrichales-Thiotrichales_incertae_sedis-Fangia 4 0 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Xanthomonadales-Sinobacteraceae-Alkanibacter 0 4 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Acidimicrobiales-Acidimicrobiaceae- Ferrimicrobium 0 2 0 0 1 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Micromonosporaceae-Asanoa 0 3 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Tsukamurellaceae-Tsukamurella 0 3 0 0 0 Bacteria-Bacteroidetes-Cytophagia-Cytophagales- Cytophagaceae-Microscilla 3 0 0 0 0 Bacteria-Chloroflexi-Caldilineae-Caldilineales- Caldilineaceae-Caldilinea 2 1 0 0 0 Bacteria-Cyanobacteria-Chloroplast-Chloroplast- Chloroplast-Chloroplast-Euglenida 0 0 0 0 3 Bacteria-Firmicutes-Bacilli-Bacillales- Anoxybacillus 0 2 1 0 0 Bacteria-Firmicutes-Bacilli-Bacillales-Gemella 0 3 0 0 0 Bacteria-Firmicutes-Bacilli-Bacillales- Planococcaceae-Jeotgalibacillus 2 0 0 0 1 Bacteria-Firmicutes-Clostridia-Clostridiales- Fusibacter 2 0 0 0 1 Bacteria-Firmicutes-Clostridia-Clostridiales- Pelotomaculum 0 3 0 0 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Ruminococcaceae-Acetivibrio 0 0 1 2 0 Bacteria-Latescibacteria- Latescibacteria_genera_incertae_sedis- Latescibacteria_genera_incertae_sedis- Latescibacteria_genera_incertae_sedis- Latescibacteria_genera_incertae_sedis 1 0 1 0 1 Bacteria-Planctomycetes-Planctomycetia- Planctomycetales-Planctomycetaceae-Singulisphaera 0 3 0 0 0 101

Bacteria-Proteobacteria-Alphaproteobacteria- Alphaproteobacteria_incertae_sedis- Rhizomicrobium-Rhizomicrobium 1 2 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Parvularculales-Parvularculaceae-Parvularcula 3 0 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Bradyrhizobiaceae-Balneimonas 0 2 1 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Bradyrhizobiaceae-Tardiphaga 0 1 1 0 1 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Brucellaceae-Paenochrobactrum 0 0 3 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Hyphomicrobiaceae- Prosthecomicrobium 3 0 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Hyphomicrobiaceae-Rhodomicrobium 1 0 0 0 2 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Rhizobiaceae-Kaistia 3 0 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Rhizobiaceae-Undefined 0 3 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Rhodobiaceae-Undefined 1 0 0 0 2 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Acetobacteraceae-Stella 0 3 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Rhodospirillaceae-Rhodospirillum 1 0 0 0 2 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Alcaligenaceae-Pusillimonas 2 0 1 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Burkholderiales_incertae_sedis- Piscinibacter 0 3 0 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Burkholderiales_incertae_sedis- Rubrivivax 2 0 0 0 1 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Schlegelella 3 0 0 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Oxalobacteraceae-Collimonas 1 0 0 0 2 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Oxalobacteraceae-Oxalobacter 0 0 0 0 3 Bacteria-Proteobacteria-Betaproteobacteria- Ferrovales-Ferrovaceae-Ferrovum 2 0 0 0 1 Bacteria-Proteobacteria-Betaproteobacteria- Neisseriales-Neisseriaceae-Formivibrio 0 0 0 0 3 Bacteria-Proteobacteria-Betaproteobacteria- Neisseriales-Neisseriaceae-Silvimonas 3 0 0 0 0 Bacteria-Proteobacteria-Deltaproteobacteria- Bdellovibrionales-Bacteriovoracaceae-Undefined 0 0 0 0 3 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfuromonadales-Undefined 3 0 0 0 0 Bacteria-Proteobacteria-Deltaproteobacteria- Myxococcales-Nannocystaceae-Undefined 1 1 0 1 0 102

Bacteria-Proteobacteria-Deltaproteobacteria- Myxococcales-Phaselicystidaceae-Phaselicystis 3 0 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Chromatiales-Chromatiaceae-Undefined 0 0 0 0 3 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae- Pectobacterium 0 0 0 0 3 Bacteria-Proteobacteria-Gammaproteobacteria- Oceanospirillales-Undefined 2 1 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Pseudomonadales-Pseudomonadaceae-Azomonas 0 0 1 0 2 Bacteria-Saccharibacteria_genera_incertae_sedis- Saccharibacteria_genera_incertae_sedis- Saccharibacteria_genera_incertae_sedis- Saccharibacteria_genera_incertae_sedis 0 1 0 1 1 Bacteria-Tenericutes-Mollicutes-Acholeplasmatales- Acholeplasmataceae-Acholeplasma 0 0 3 0 0 Archaea-Euryarchaeota-Methanomicrobia- Undefined 0 0 0 0 2 Archaea-Euryarchaeota-Thermoplasmata- Thermoplasmatales-Undefined 0 0 1 1 0 Archaea-Thaumarchaeota-Nitrosopumilales- Nitrosopumilales-Nitrosopumilaceae- Nitrosopumilus 0 2 0 0 0 Bacteria-Acidobacteria-Acidobacteria_Gp5-Gp5- Gp5-Gp5 1 0 1 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Acidothermaceae-Acidothermus 0 2 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Beutenbergiaceae-Undefined 0 0 2 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Dermatophilaceae-Austwickia 2 0 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Intrasporangiaceae-Tetrasphaera 0 2 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Kineosporiaceae-Angustibacter 0 1 1 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Microbacteriaceae-Rhodoglobus 2 0 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Micromonosporaceae- Virgisporangium 0 1 1 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Streptosporangiaceae-Nonomuraea 0 2 0 0 0 Bacteria-Armatimonadetes-Undefined 2 0 0 0 0 Bacteria-Bacteroidetes-Bacteroidia-Bacteroidales- Marinilabiliaceae-Undefined 0 0 0 0 2 Bacteria-Bacteroidetes-Cytophagia-Cytophagales- Cytophagaceae-Larkinella 0 1 0 1 0 Bacteria-Bacteroidetes-Cytophagia-Cytophagales- Cytophagaceae-Siphonobacter 2 0 0 0 0 Bacteria-Bacteroidetes-Flavobacteriia- Flavobacteriales-Flavobacteriaceae-Elizabethkingia 2 0 0 0 0 103

Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Chitinophagaceae-Niabella 0 2 0 0 0 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Chitinophagaceae-Niastella 1 0 0 1 0 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Rhodothermaceae-Undefined 1 1 0 0 0 Bacteria-Chloroflexi-Chloroflexia-Chloroflexales- Chloroflexaceae-Roseiflexus 0 1 1 0 0 Bacteria-Chloroflexi-Chloroflexia-Chloroflexales- Chloroflexaceae-Undefined 0 2 0 0 0 Bacteria-Cloacimonetes-Undefined 1 0 0 1 0 Bacteria-Firmicutes-Bacilli-Bacillales- Alicyclobacillaceae-Alicyclobacillus 0 2 0 0 0 Bacteria-Firmicutes-Bacilli-Bacillales-Fontibacillus 0 1 1 0 0 Bacteria-Firmicutes-Bacilli-Bacillales-Geobacillus 1 0 0 1 0 Bacteria-Firmicutes-Bacilli-Bacillales- Planococcaceae-Ureibacillus 0 0 2 0 0 Bacteria-Firmicutes-Bacilli-Bacillales- Staphylococcaceae-Undefined 0 2 0 0 0 Bacteria-Firmicutes-Bacilli-Lactobacillales- Undefined 0 1 0 1 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Clostridiisalibacter 0 0 0 2 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Dehalobacter 0 0 0 2 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Desulfitobacterium 0 0 0 2 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Fervidicella 0 0 0 2 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Gracilibacteraceae-Gracilibacter 0 0 0 2 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Proteiniclasticum 0 0 1 1 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Ruminococcaceae-Sporobacter 0 0 0 1 1 Bacteria-Firmicutes-Clostridia-Clostridiales- Sedimentibacter 0 0 2 0 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Syntrophomonadaceae-Syntrophomonas 0 0 0 2 0 Bacteria-Firmicutes-Erysipelotrichia- Erysipelotrichales-Erysipelotrichaceae-Turicibacter 1 0 0 1 0 Bacteria-Firmicutes-Negativicutes- Selenomonadales-Veillonellaceae-Pelosinus 0 0 0 0 2 Bacteria-Firmicutes-Negativicutes- Selenomonadales-Veillonellaceae-Propionispira 0 0 0 1 1 Bacteria-Lentisphaerae-Oligosphaeria- Oligosphaerales-Oligosphaeraceae-Oligosphaera 1 0 0 0 1 Bacteria-Proteobacteria-Alphaproteobacteria- Caulobacterales-Undefined 0 0 2 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Bradyrhizobiaceae-Salinarimonas 0 1 1 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Phyllobacteriaceae-Phyllobacterium 0 0 2 0 0 104

Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Xanthobacteraceae-Ancylobacter 0 2 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Xanthobacteraceae-Labrys 1 1 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Rhodospirillaceae-Defluviicoccus 0 2 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Rhodospirillaceae-Dongia 0 2 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Sphingomonadales-Erythrobacteraceae- Croceicoccus 0 0 0 0 2 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Burkholderiaceae-Burkholderia 0 0 0 0 2 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Giesbergeria 0 0 0 0 2 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Xenophilus 0 0 1 0 1 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Oxalobacteraceae-Rugamonas 0 0 2 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Neisseriales-Neisseriaceae-Chitinilyticum 1 0 0 0 1 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfobacterales-Desulfobacteraceae-Desulfatitalea 0 2 0 0 0 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfobacterales-Undefined 0 0 0 0 2 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfovibrionales-Undefined 0 0 2 0 0 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfuromonadales-Desulfuromonadaceae- Undefined 2 0 0 0 0 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfuromonadales-Geobacteraceae- Geopsychrobacter 1 0 0 0 1 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfuromonadales-Geobacteraceae-Undefined 0 2 0 0 0 Bacteria-Proteobacteria-Deltaproteobacteria- Myxococcales-Cystobacteraceae-Cystobacter 0 1 0 0 1 Bacteria-Proteobacteria-Deltaproteobacteria- Myxococcales-Cystobacteraceae-Stigmatella 0 1 0 0 1 Bacteria-Proteobacteria-Deltaproteobacteria- Syntrophobacterales-Syntrophobacteraceae- Syntrophobacter 2 0 0 0 0 Bacteria-Proteobacteria-Deltaproteobacteria- Syntrophobacterales-Undefined 0 1 0 0 1 Bacteria-Proteobacteria-Gammaproteobacteria- Alteromonadales-Undefined 0 0 0 0 2 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Gibbsiella 0 2 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Lonsdalea 2 0 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Serratia 0 0 2 0 0 105

Bacteria-Proteobacteria-Gammaproteobacteria- Legionellales-Coxiellaceae-Undefined 2 0 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Legionellales-Undefined 2 0 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Methylococcales-Methylococcaceae-Methylococcus 2 0 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Pseudomonadales-Pseudomonadaceae-Azotobacter 0 0 0 0 2 Bacteria-Proteobacteria-Gammaproteobacteria- Xanthomonadales-Xanthomonadaceae-Fulvimonas 0 1 0 0 1 Bacteria-Spirochaetes-Spirochaetia-Spirochaetales- Spirochaetaceae-Treponema 1 0 0 0 1 Bacteria-Spirochaetes-Spirochaetia-Spirochaetales- Spirochaetaceae-Undefined 1 0 0 0 1 Bacteria-Synergistetes-Synergistia-Synergistales- Synergistaceae-Thermovirga 0 0 2 0 0 Bacteria-Verrucomicrobia-Spartobacteria- Xiphinematobacter-Xiphinematobacter- Xiphinematobacter 1 1 0 0 0 Archaea-Euryarchaeota-Archaeoglobi- Archaeoglobales-Archaeoglobaceae-Ferroglobus 1 0 0 0 0 Archaea-Euryarchaeota-Methanobacteria- Methanobacteriales-Methanobacteriaceae- Methanobacterium 1 0 0 0 0 Archaea-Euryarchaeota-Methanobacteria- Methanobacteriales-Methanobacteriaceae- Methanobrevibacter 0 0 0 0 1 Archaea-Euryarchaeota-Methanomicrobia- Methanomicrobiales- Methanomicrobiales_incertae_sedis-Methanoregula 1 0 0 0 0 Archaea-Euryarchaeota-Methanomicrobia- Methanomicrobiales-Methanospirillaceae- Methanospirillum 1 0 0 0 0 Bacteria-Acidobacteria-Acidobacteria_Gp13-Gp13- Gp13-Gp13 0 1 0 0 0 Bacteria-Acidobacteria-Acidobacteria_Gp25-Gp25- Gp25-Gp25 0 0 1 0 0 Bacteria-Acidobacteria-Holophagae-Holophagales- Holophagaceae-Holophaga 1 0 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Beutenbergiaceae-Miniimonas 0 1 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Cellulomonadaceae-Actinotalea 0 1 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Cellulomonadaceae- Sediminihabitans 0 1 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Demequinaceae-Lysinimicrobium 0 0 0 0 1 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Dermacoccaceae-Undefined 0 0 1 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Intrasporangiaceae-Marihabitans 1 0 0 0 0 106

Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Intrasporangiaceae-Ornithinibacter 0 1 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Jiangellaceae-Jiangella 0 0 1 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Microbacteriaceae-Salinibacterium 0 0 1 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Microbacteriaceae-Subtercola 1 0 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Micrococcineae_incertae_sedis- Luteimicrobium 0 1 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Micromonosporaceae- Catellatospora 0 1 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Micromonosporaceae-Salinispora 0 1 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Propionibacteriaceae-Propioniferax 1 0 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Pseudonocardiaceae-Umezawaea 0 0 1 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Ruaniaceae-Ruania 0 1 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Thermomonosporaceae- Actinoallomurus 0 0 1 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Thermomonosporaceae- Thermomonospora 1 0 0 0 0 Bacteria-Actinobacteria-Actinobacteria- Actinomycetales-Thermomonosporaceae-Undefined 1 0 0 0 0 Bacteria-BRC1-BRC1_genera_incertae_sedis- BRC1_genera_incertae_sedis- BRC1_genera_incertae_sedis- BRC1_genera_incertae_sedis 0 0 1 0 0 Bacteria-Bacteroidetes-Bacteroidia-Bacteroidales- Porphyromonadaceae-Odoribacter 1 0 0 0 0 Bacteria-Bacteroidetes-Bacteroidia-Bacteroidales- Porphyromonadaceae-Porphyromonas 0 1 0 0 0 Bacteria-Bacteroidetes-Bacteroidia-Bacteroidales- Prevotellaceae-Paraprevotella 0 0 0 0 1 Bacteria-Bacteroidetes-Bacteroidia-Bacteroidales- Prevotellaceae-Undefined 0 0 0 0 1 Bacteria-Bacteroidetes-Bacteroidia-Bacteroidales- Prevotellaceae-Xylanibacter 0 0 0 0 1 Bacteria-Bacteroidetes-Cytophagia-Cytophagales- Cytophagaceae-Leadbetterella 0 0 0 0 1 Bacteria-Bacteroidetes-Cytophagia-Cytophagales- Cytophagaceae-Rhodocytophaga 0 1 0 0 0 Bacteria-Bacteroidetes-Cytophagia-Cytophagales- Cytophagaceae-Rudanella 0 0 0 0 1 Bacteria-Bacteroidetes-Cytophagia-Cytophagales- Mooreiaceae-Mooreia 0 1 0 0 0 107

Bacteria-Bacteroidetes-Flavobacteriia- Flavobacteriales-Cryomorphaceae-Crocinitomix 0 0 0 0 1 Bacteria-Bacteroidetes-Flavobacteriia- Flavobacteriales-Flavobacteriaceae-Lutibacter 0 0 0 0 1 Bacteria-Bacteroidetes-Flavobacteriia- Flavobacteriales-Flavobacteriaceae-Soonwooa 0 0 1 0 0 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Chitinophagaceae-Filimonas 0 0 0 0 1 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Cyclobacteriaceae-Belliella 0 0 1 0 0 Bacteria-Bacteroidetes-Sphingobacteriia- Sphingobacteriales-Saprospiraceae-Aureispira 0 0 0 0 1 Bacteria-Chlamydiae-Chlamydiia-Chlamydiales- Chlamydiaceae-Chlamydophila 1 0 0 0 0 Bacteria-Chlamydiae-Chlamydiia-Chlamydiales- Chlamydiaceae-Undefined 1 0 0 0 0 Bacteria-Chlorobi-Chlorobia-Chlorobiales- Chlorobiaceae-Chlorobium 1 0 0 0 0 Bacteria-Chloroflexi-Caldilineae-Caldilineales- Caldilineaceae-Undefined 0 0 0 0 1 Bacteria-Chloroflexi-Chloroflexia-Chloroflexales- Chloroflexaceae-Heliothrix 1 0 0 0 0 Bacteria-Chloroflexi-Dehalococcoidetes- Dehalogenimonas-Dehalogenimonas- Dehalogenimonas 1 0 0 0 0 Bacteria-Chloroflexi-Ktedonobacteria-Undefined 0 1 0 0 0 Bacteria-Cyanobacteria-Chloroplast-Cyanobacteria- GpIIa 0 0 0 1 0 Bacteria-Cyanobacteria-Chloroplast-Cyanobacteria- GpVIII 1 0 0 0 0 Bacteria-Cyanobacteria-Chloroplast-Cyanobacteria- GpX 1 0 0 0 0 Bacteria-Deinococcus-Thermus-Deinococci- Deinococcales-Deinococcaceae-Deinobacterium 0 1 0 0 0 Bacteria-Deinococcus-Thermus-Deinococci- Deinococcales-Deinococcaceae-Undefined 0 1 0 0 0 Bacteria-Fibrobacteres-Fibrobacteria- Fibrobacterales-Fibrobacteraceae-Fibrobacter 0 0 0 0 1 Bacteria-Firmicutes-Bacilli-Bacillales-Aeribacillus 0 0 0 1 0 Bacteria-Firmicutes-Bacilli-Bacillales- Alicyclobacillaceae-Undefined 0 1 0 0 0 Bacteria-Firmicutes-Bacilli-Bacillales-Domibacillus 0 0 0 0 1 Bacteria-Firmicutes-Bacilli-Bacillales-Falsibacillus 0 0 1 0 0 Bacteria-Firmicutes-Bacilli-Bacillales-Halobacillus 0 0 0 1 0 Bacteria-Firmicutes-Bacilli-Bacillales-Lentibacillus 0 1 0 0 0 Bacteria-Firmicutes-Bacilli-Bacillales- Ornithinibacillus 0 0 1 0 0 Bacteria-Firmicutes-Bacilli-Bacillales-Planifilum 0 1 0 0 0 Bacteria-Firmicutes-Bacilli-Bacillales- Planococcaceae-Kurthia 0 1 0 0 0 Bacteria-Firmicutes-Bacilli-Bacillales- Planococcaceae-Rummeliibacillus 0 0 1 0 0 108

Bacteria-Firmicutes-Bacilli-Bacillales- Planococcaceae-Solibacillus 1 0 0 0 0 Bacteria-Firmicutes-Bacilli-Bacillales- Thermoactinomyces 0 1 0 0 0 Bacteria-Firmicutes-Bacilli-Bacillales-Virgibacillus 0 1 0 0 0 Bacteria-Firmicutes-Bacilli-Lactobacillales- Aerococcaceae-Aerococcus 0 0 0 0 1 Bacteria-Firmicutes-Bacilli-Lactobacillales- Carnobacteriaceae-Desemzia 0 1 0 0 0 Bacteria-Firmicutes-Bacilli-Lactobacillales- Carnobacteriaceae-Isobaculum 0 0 0 1 0 Bacteria-Firmicutes-Bacilli-Lactobacillales- Carnobacteriaceae-Trichococcus 0 0 0 0 1 Bacteria-Firmicutes-Bacilli-Lactobacillales- Enterococcaceae-Vagococcus 0 0 1 0 0 Bacteria-Firmicutes-Bacilli-Lactobacillales- Lactobacillaceae-Lactobacillus 0 0 0 0 1 Bacteria-Firmicutes-Bacilli-Lactobacillales- Leuconostocaceae-Leuconostoc 0 0 1 0 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Alkaliphilus 0 0 0 1 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Caldicoprobacter 0 1 0 0 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Finegoldia 0 1 0 0 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Geosporobacter 0 0 0 1 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Gracilibacteraceae-Lutispora 0 0 0 1 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Lachnospiraceae-Coprococcus 1 0 0 0 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Lachnospiraceae-Roseburia 0 0 0 0 1 Bacteria-Firmicutes-Clostridia-Clostridiales- Peptostreptococcaceae-Sporacetigenium 0 0 1 0 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Ruminococcaceae-Acetanaerobacterium 0 0 0 1 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Ruminococcaceae-Anaerotruncus 1 0 0 0 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Ruminococcaceae-Ethanoligenens 0 0 0 1 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Ruminococcaceae-Ruminococcus 0 0 0 1 0 Bacteria-Firmicutes-Clostridia-Clostridiales- Syntrophobotulus 0 0 0 1 0 Bacteria-Firmicutes-Clostridia-Halanaerobiales- Halobacteroidaceae-Undefined 0 0 0 1 0 Bacteria-Firmicutes-Clostridia-Natranaerobiales- Natranaerobiaceae-Undefined 0 0 1 0 0 Bacteria-Firmicutes-Clostridia- Thermoanaerobacterales-Thermoanaerobacteraceae- Undefined 0 0 1 0 0 109

Bacteria-Firmicutes-Clostridia- Thermoanaerobacterales-Undefined 0 0 1 0 0 Bacteria-Firmicutes-Erysipelotrichia- Erysipelotrichales-Erysipelotrichaceae-Holdemania 0 0 1 0 0 Bacteria-Firmicutes-Negativicutes- Selenomonadales-Acidaminococcaceae-Succinispira 0 0 0 0 1 Bacteria-Firmicutes-Negativicutes- Selenomonadales-Veillonellaceae-Psychrosinus 0 0 0 0 1 Bacteria-Hydrogenedentes-Undefined 0 1 0 0 0 Bacteria-Planctomycetes-Planctomycetia- Planctomycetales-Planctomycetaceae-Telmatocola 1 0 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Caulobacterales-Hyphomonadaceae-Undefined 1 0 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Aurantimonadaceae-Aureimonas 0 1 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Brucellaceae-Brucella 0 1 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Brucellaceae-Pseudochrobactrum 0 1 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Brucellaceae-Undefined 0 1 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Rhodobiaceae-Anderseniella 0 1 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Rhodobiaceae-Rhodobium 1 0 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Xanthobacteraceae-Azorhizobium 0 1 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhizobiales-Xanthobacteraceae-Xanthobacter 0 0 0 1 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodobacterales-Rhodobacteraceae-Hwanghaeicola 0 1 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodobacterales-Rhodobacteraceae-Phaeobacter 0 0 0 1 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodobacterales-Rhodobacteraceae-Roseivivax 1 0 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Elioraea-Elioraea 1 0 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Rhodospirillaceae-Desertibacter 0 1 0 0 0 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Rhodospirillaceae-Insolitispirillum 0 0 0 0 1 Bacteria-Proteobacteria-Alphaproteobacteria- Rhodospirillales-Rhodospirillaceae-Phaeospirillum 0 0 0 0 1 Bacteria-Proteobacteria-Alphaproteobacteria- Sphingomonadales-Sphingomonadaceae-Stakelama 0 1 0 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Alcaligenaceae-Alcaligenes 0 1 0 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Burkholderiaceae-Ralstonia 1 0 0 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Burkholderiales_incertae_sedis- Paucibacter 0 0 1 0 0 110

Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Burkholderiales_incertae_sedis- Thiomonas 0 0 0 0 1 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Ottowia 0 0 0 0 1 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Comamonadaceae-Tepidicella 0 1 0 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Burkholderiales-Sutterellaceae-Parasutterella 1 0 0 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Procabacteriales-Procabacteriaceae-Undefined 0 1 0 0 0 Bacteria-Proteobacteria-Betaproteobacteria- Rhodocyclales-Rhodocyclaceae-Azovibrio 0 0 0 0 1 Bacteria-Proteobacteria-Betaproteobacteria- Rhodocyclales-Rhodocyclaceae-Denitratisoma 0 0 0 0 1 Bacteria-Proteobacteria-Betaproteobacteria- Rhodocyclales-Rhodocyclaceae-Georgfuchsia 0 0 0 0 1 Bacteria-Proteobacteria-Betaproteobacteria- Rhodocyclales-Rhodocyclaceae-Sterolibacterium 0 1 0 0 0 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfobacterales-Desulfobacteraceae- Desulfobacterium 0 0 0 0 1 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfobacterales-Desulfobacteraceae- Desulfoconvexum 0 0 0 0 1 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfobacterales-Desulfobulbaceae-Desulfobulbus 0 0 0 0 1 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfobacterales-Desulfobulbaceae- Desulforhopalus 0 0 0 0 1 Bacteria-Proteobacteria-Deltaproteobacteria- Desulfovibrionales-Desulfovibrionaceae-Undefined 1 0 0 0 0 Bacteria-Proteobacteria-Deltaproteobacteria- Myxococcales-Kofleriaceae-Kofleria 0 1 0 0 0 Bacteria-Proteobacteria-Deltaproteobacteria- Myxococcales-Myxococcaceae-Pyxidicoccus 0 1 0 0 0 Bacteria-Proteobacteria-Deltaproteobacteria- Myxococcales-Polyangiaceae-Jahnella 1 0 0 0 0 Bacteria-Proteobacteria-Deltaproteobacteria- Syntrophobacterales-Syntrophaceae-Desulfobacca 0 0 0 0 1 Bacteria-Proteobacteria-Deltaproteobacteria- Syntrophobacterales-Syntrophaceae-Syntrophus 1 0 0 0 0 Bacteria-Proteobacteria-Deltaproteobacteria- Syntrophobacterales-Syntrophobacteraceae- Desulforhabdus 0 0 0 0 1 Bacteria-Proteobacteria-Deltaproteobacteria- Syntrophobacterales-Syntrophobacteraceae- Undefined 0 0 0 0 1 Bacteria-Proteobacteria-Deltaproteobacteria- Syntrophorhabdaceae-Syntrophorhabdaceae- Syntrophorhabdus 0 0 0 0 1 Bacteria-Proteobacteria-Epsilonproteobacteria- Campylobacterales-Helicobacteraceae-Helicobacter 0 0 0 1 0 111

Bacteria-Proteobacteria-Epsilonproteobacteria- Campylobacterales-Helicobacteraceae-Sulfurovum 0 0 0 0 1 Bacteria-Proteobacteria-Epsilonproteobacteria- Nautiliales-Nautiliaceae-Nitratiruptor 0 0 0 0 1 Bacteria-Proteobacteria-Gammaproteobacteria- Aeromonadales-Undefined 0 0 0 0 1 Bacteria-Proteobacteria-Gammaproteobacteria- Alteromonadales-Alteromonadaceae-Undefined 1 0 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Chromatiales-Chromatiaceae-Thiobaca 0 0 0 0 1 Bacteria-Proteobacteria-Gammaproteobacteria- Chromatiales-Chromatiaceae-Thiohalocapsa 0 0 0 0 1 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Budvicia 0 0 0 0 1 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Kosakonia 0 0 0 0 1 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Lelliottia 0 1 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae- Obesumbacterium 1 0 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Rahnella 0 0 0 0 1 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Raoultella 0 0 0 0 1 Bacteria-Proteobacteria-Gammaproteobacteria- Enterobacteriales-Enterobacteriaceae-Trabulsiella 0 1 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Gammaproteobacteria_incertae_sedis-Gilvimarinus- Gilvimarinus 0 1 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Methylococcales-Methylococcaceae-Methylocaldum 0 0 0 0 1 Bacteria-Proteobacteria-Gammaproteobacteria- Methylococcales-Methylococcaceae- Methylomarinum 0 0 0 0 1 Bacteria-Proteobacteria-Gammaproteobacteria- Methylococcales-Methylococcaceae-Methylosoma 0 0 0 0 1 Bacteria-Proteobacteria-Gammaproteobacteria- Oceanospirillales-Halomonadaceae-Undefined 1 0 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Oceanospirillales-Oleiphilaceae-Oleiphilus 0 0 0 0 1 Bacteria-Proteobacteria-Gammaproteobacteria- Orbales-Orbaceae-Undefined 1 0 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Pseudomonadales-Pseudomonadaceae- Azorhizophilus 1 0 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Thiotrichales-Undefined 1 0 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Vibrionales-Vibrionaceae-Vibrio 1 0 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Xanthomonadales-Xanthomonadaceae-Dokdonella 0 0 0 1 0 112

Bacteria-Proteobacteria-Gammaproteobacteria- Xanthomonadales-Xanthomonadaceae-Dyella 0 0 1 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Xanthomonadales-Xanthomonadaceae- Thermomonas 0 1 0 0 0 Bacteria-Proteobacteria-Gammaproteobacteria- Xanthomonadales-Xanthomonadaceae-Xylella 0 0 1 0 0 Bacteria-Spirochaetes-Spirochaetia-Spirochaetales- Brachyspiraceae-Brachyspira 1 0 0 0 0 Bacteria-Spirochaetes-Spirochaetia-Spirochaetales- Spirochaetaceae-Borrelia 0 0 0 0 1 Bacteria-Spirochaetes-Spirochaetia-Spirochaetales- Spirochaetales_incertae_sedis-Exilispira 0 0 0 0 1 Bacteria-Synergistetes-Synergistia-Synergistales- Synergistaceae-Undefined 0 0 1 0 0 Bacteria-Verrucomicrobia-Verrucomicrobiae- Verrucomicrobiales-Undefined 0 0 0 0 1 Bacteria-Verrucomicrobia-Verrucomicrobiae- Verrucomicrobiales-Verrucomicrobiaceae- Persicirhabdus 1 0 0 0 0 Bacteria-Verrucomicrobia-Verrucomicrobiae- Verrucomicrobiales-Verrucomicrobiaceae- Verrucomicrobium 1 0 0 0 0

APPENDIX B

16S rDNA PCA data

114

16S rDNA PCA Rotation PC1 PC2 PC3 PC4 PC5 PC1 PC2 PC3 PC4 PC5 [1,] -0.4521 0.0532 0.0040 0.0365 0.0141 [45,] -0.4428 0.0516 -0.0027 0.0319 0.0128 [2,] -0.4520 0.0532 0.0041 0.0366 0.0142 [46,] -0.4520 0.0531 0.0042 0.0365 0.0140 [3,] -0.4518 0.0533 0.0040 0.0367 0.0143 [47,] 0.2749 -0.6853 1.0064 0.1899 -0.2891 [4,] -0.4520 0.0530 0.0041 0.0365 0.0141 [48,] -0.4168 0.0672 0.0022 0.0382 0.0257 [5,] -0.4476 0.0545 0.0040 0.0403 0.0178 [49,] -0.4396 0.0521 0.0108 0.0429 0.0080 [6,] -0.4520 0.0533 0.0041 0.0362 0.0142 [50,] -0.4366 0.0542 -0.0007 0.0189 0.0168 [7,] -0.4499 0.0544 0.0047 0.0333 0.0145 [51,] -0.4520 0.0532 0.0041 0.0366 0.0143 [8,] -0.4507 0.0536 0.0040 0.0377 0.0152 [52,] 1.4386 1.1085 0.2498 -2.9689 0.6265 [9,] -0.4504 0.0526 0.0047 0.0369 0.0138 [53,] 0.0043 -0.6432 -0.7851 -0.0942 -0.0835 [10,] -0.4516 0.0534 0.0039 0.0364 0.0140 [54,] -0.4487 0.0493 -0.0001 0.0359 0.0140 [11,] -0.4520 0.0532 0.0041 0.0363 0.0142 [55,] -0.4504 0.0530 0.0043 0.0376 0.0148 [12,] -0.4465 0.0559 0.0046 0.0296 0.0154 [56,] 2.0497 -4.2959 -5.0210 -0.8739 -0.6259 [13,] -0.4511 0.0535 0.0041 0.0358 0.0147 [57,] -0.4383 0.0561 0.0044 0.0485 0.0289 [14,] -0.4515 0.0529 0.0047 0.0367 0.0138 [58,] -0.4478 0.0555 0.0038 0.0320 0.0144 [15,] -0.4521 0.0531 0.0039 0.0364 0.0141 [59,] -0.4520 0.0532 0.0041 0.0363 0.0142 [16,] -0.4465 0.0552 0.0051 0.0319 0.0131 [60,] -0.4514 0.0535 0.0041 0.0365 0.0145 [17,] -0.4354 0.0613 0.0046 0.0128 0.0188 [61,] -0.4519 0.0531 0.0039 0.0366 0.0142 [18,] -0.4520 0.0531 0.0039 0.0365 0.0141 [62,] -0.4518 0.0533 0.0040 0.0367 0.0143 [19,] -0.4432 0.0567 0.0037 0.0283 0.0160 [63,] -0.4520 0.0532 0.0041 0.0366 0.0142 [20,] -0.4502 0.0539 0.0041 0.0339 0.0144 [64,] -0.4505 0.0534 0.0041 0.0380 0.0161 [21,] -0.4515 0.0534 0.0040 0.0370 0.0145 [65,] -0.4424 0.0592 0.0058 0.0211 0.0174 [22,] -0.4519 0.0534 0.0041 0.0362 0.0141 [66,] -0.4215 0.0689 0.0005 0.0613 0.0190 [23,] 10.1133 6.3652 -1.2200 0.1004 -0.6532 [67,] -0.4268 0.0669 0.0043 0.0274 0.0074 [24,] -0.4507 0.0539 0.0044 0.0345 0.0147 [68,] 3.0616 -0.3326 1.2510 -2.5768 0.1398 [25,] -0.4505 0.0540 0.0042 0.0343 0.0147 [69,] 11.7756 -3.9955 0.7526 1.1349 0.9209 [26,] -0.4519 0.0531 0.0038 0.0365 0.0139 [70,] -0.3995 0.0785 0.0031 0.0152 0.0217 [27,] -0.4506 0.0534 0.0048 0.0356 0.0139 [71,] -0.4205 0.0592 0.0049 0.0654 0.0488 [28,] -0.4401 0.0564 0.0041 0.0462 0.0250 [72,] 2.7979 -2.9092 3.4210 -0.3251 -1.3443 [29,] -0.4520 0.0533 0.0040 0.0365 0.0140 [73,] -0.3581 0.0624 0.0123 0.0701 0.0182 [30,] -0.4521 0.0531 0.0039 0.0364 0.0141 [74,] -0.4506 0.0543 0.0037 0.0374 0.0134 [31,] -0.4493 0.0555 0.0036 0.0375 0.0120 [75,] -0.4519 0.0531 0.0042 0.0364 0.0143 [32,] -0.4404 0.0533 0.0003 0.0426 0.0185 [76,] -0.4515 0.0534 0.0040 0.0370 0.0145 [33,] 0.2297 0.4174 -0.0629 0.5426 -0.0164 [77,] -0.4520 0.0529 0.0037 0.0364 0.0141 [34,] 0.5826 0.4484 -0.0948 0.8216 0.4350 [78,] -0.4520 0.0529 0.0037 0.0364 0.0141 [35,] 0.9988 0.9096 -0.2187 1.0749 -0.0830 [79,] 0.3054 -0.5853 0.4584 0.0659 -0.2634 [36,] -0.2982 0.0796 -0.0088 0.1548 0.1303 [80,] -0.4217 0.0759 0.0007 0.0352 0.0029 [37,] -0.4441 0.0586 0.0041 0.0353 0.0101 [81,] -0.4245 0.0676 0.0058 0.0498 -0.0162 [38,] -0.4521 0.0532 0.0040 0.0365 0.0141 [82,] -0.4488 0.0549 0.0038 0.0374 0.0142 [39,] -0.4467 0.0548 0.0040 0.0387 0.0175 [83,] -0.4516 0.0535 0.0040 0.0368 0.0141 [40,] -0.4511 0.0536 0.0039 0.0366 0.0144 [84,] -0.4505 0.0544 0.0039 0.0368 0.0131 [41,] -0.4499 0.0549 0.0038 0.0354 0.0130 [85,] -0.4369 0.0620 0.0011 0.0465 0.0123 [42,] -0.4521 0.0532 0.0040 0.0365 0.0141 [86,] -0.4514 0.0532 0.0040 0.0362 0.0141 [43,] -0.4515 0.0529 0.0044 0.0363 0.0141 [87,] -0.4499 0.0537 0.0032 0.0354 0.0132 [44,] -0.4253 0.0204 -0.0368 0.0192 0.0108 [88,] -0.4331 0.0389 0.0243 0.0428 0.0123 115

16S rDNA PCA Loadings PC1 PC2 PC3 PC4 PC5 Creosote Marsh 0.4650 0.4931 -0.1716 0.3174 -0.6407 Big Marsh Well 0.4830 0.3091 0.0950 -0.7961 0.1685 Slag Dune SW 0.3472 -0.6049 -0.6997 -0.1266 -0.0889 Big Marsh SW 0.3829 -0.5389 0.6864 0.0414 -0.3002 RR Ditch 0.5324 0.0710 0.0263 0.4977 0.6806

16S rDNA Importance of PCA Components PC1 PC2 PC3 PC4 PC5 Standard deviation 1.797 1.002 0.699 0.4786 0.2218 Proportion of Variance 0.646 0.201 0.098 0.0458 0.0098 Cumulative Proportion 0.646 0.847 0.944 0.9902 1

APPENDIX C

PCA DATA Biologs

117

PCA Summary for BIOLOG Data PC1 PC2 PC3 PC4 PC5 PC6 PC7 PC8 PC9 PC10 PC11 Standard deviation 4.19 1.83 1.61 1.48 1.27 1.12 0.96 0.77 0.73 0.66 0.00 Proportion of Varience 0.57 0.11 0.08 0.07 0.05 0.04 0.03 0.02 0.02 0.01 0.00 Cumulative Proportion 0.57 0.67 0.76 0.83 0.88 0.92 0.95 0.97 0.99 1.00 1.00

BIOLOG PCA Loadings

PC1 PC2 PC3 PC4 PC5 PC6 PC7 PC8 PC9 PC10 PC11 [1,] -1.585 -2.800 -1.668 1.208 -0.447 2.209 -0.461 0.477 -0.222 0.234 0.000 [2,] 2.186 -1.904 3.029 -0.924 -0.405 -0.112 0.713 -0.077 -0.070 1.177 0.000 [3,] -2.185 2.001 -1.217 -0.248 2.236 -0.465 -0.177 0.559 -0.839 0.909 0.000 [4,] -0.540 0.315 -1.212 -0.937 1.159 0.758 0.605 -1.585 1.077 0.067 0.000 [5,] -1.637 -2.125 1.278 -1.550 1.444 -0.580 -1.338 0.172 -0.016 -1.009 0.000 [6,] 6.144 2.867 1.624 0.246 0.227 1.683 0.044 0.420 -0.070 -0.604 0.000 [7,] 7.359 -1.078 -1.037 2.441 0.353 -1.646 -0.124 -0.047 0.344 -0.045 0.000 [8,] -5.608 1.242 1.462 2.156 -0.974 -0.312 -0.675 -1.112 -0.581 -0.098 0.000 [9,] -5.701 0.819 0.361 0.561 -0.514 -0.529 0.751 1.222 1.326 -0.126 0.000 [10,] 2.013 1.233 -1.524 -2.095 -2.322 -0.511 -1.262 0.023 0.121 0.295 0.000 [11,] -0.446 -0.570 -1.096 -0.860 -0.758 -0.497 1.924 -0.052 -1.072 -0.800 0.000

118

Biolog PCA Rotation

PC1 PC2 PC3 PC4 PC5 PC6 PC7 PC8 PC9 PC10 PC11

C1 0.1788 -0.1731 -0.2140 0.1399 -0.2442 0.2156 0.1538 -0.0087 -0.0568 0.0220 -0.0113

C2 0.1837 0.2907 0.0225 -0.1147 0.0234 0.1941 0.2071 -0.0572 -0.0792 -0.0873 -0.2371

C3 0.2133 0.2055 -0.0876 0.0833 0.0602 -0.0750 0.0063 -0.0751 0.1392 0.0069 0.2553

C4 0.1611 -0.3052 -0.0869 0.0180 0.2580 0.1451 -0.2545 0.1386 -0.1201 0.0102 -0.2443

C5 0.2051 -0.1585 0.0743 0.0121 -0.2239 0.1840 0.1466 0.1080 -0.1374 0.1037 0.2499

C6 0.2102 0.1112 0.0854 -0.2287 0.1299 -0.0920 -0.0734 -0.0848 0.0082 -0.1051 -0.3991

C7 0.1592 0.2674 -0.2480 -0.0390 0.1247 -0.1696 0.2856 -0.0885 0.1439 0.0514 0.1989

C8 0.2293 0.1086 0.0018 0.0706 -0.0458 0.0210 -0.0567 -0.0403 -0.1978 0.0192 -0.2277

C9 0.1959 0.2022 0.0468 0.1923 -0.0262 0.0720 -0.1208 0.1839 0.1166 -0.3561 -0.2441

C10 0.1016 0.1601 0.4561 0.0227 -0.0560 0.3520 0.1132 0.0343 -0.1482 0.1255 -0.0497

C11 0.1724 -0.0970 0.3079 0.1450 0.0010 -0.1157 0.1803 0.0483 0.0858 0.4843 -0.1407

C12 0.2110 -0.0877 0.0990 -0.1205 0.1710 -0.1441 -0.1141 -0.2498 -0.0682 -0.1688 0.0704

C13 0.1705 -0.2302 -0.0465 -0.2720 0.1769 0.0142 0.1461 -0.3488 -0.0218 -0.0844 0.0552

C14 0.1922 0.0990 -0.0225 -0.1304 0.2841 -0.2839 0.0498 -0.0072 -0.2166 0.2328 0.1822

C15 0.1672 0.0849 -0.2709 0.2839 0.0518 0.1549 -0.0623 -0.2628 -0.1193 0.2732 -0.1326

C16 0.1894 -0.2300 0.1409 -0.0614 -0.1357 -0.1902 -0.1824 0.1875 -0.0949 -0.0838 -0.0687

C17 0.1478 -0.3354 -0.2259 0.1311 -0.0531 0.1582 -0.0459 0.0668 -0.1938 -0.1477 0.2036 - C18 0.1062 0.1229 0.1862 0.3286 -0.2002 -0.0832 -0.2421 -0.6147 -0.3638 -0.0735 0.0759

C19 0.1883 0.1559 0.0768 0.2829 0.0868 0.0632 -0.0176 0.1693 0.1109 -0.3893 0.1651

C20 0.1730 -0.2795 0.1312 -0.2278 -0.0794 -0.1234 -0.0166 -0.0763 -0.0606 -0.2185 0.1555

C21 0.2163 0.0264 -0.1350 -0.1580 0.1594 0.1433 -0.0553 -0.0684 -0.0911 -0.0142 -0.1273

C22 0.1724 0.1728 0.1672 -0.2310 -0.0715 0.2238 -0.1968 -0.1680 0.3438 -0.0977 0.2564

C23 0.2162 0.0790 0.0690 0.1322 -0.1545 -0.1471 -0.1397 0.1140 0.1655 0.0971 0.1630

C24 0.1925 -0.0962 -0.0363 -0.1393 -0.3087 -0.0491 0.3131 -0.1113 0.1878 0.0436 -0.2271

C25 0.1983 -0.0429 -0.2261 0.0920 -0.2198 0.1751 -0.0256 -0.1435 0.2144 0.0090 -0.0016

C26 0.1393 -0.1066 -0.1320 0.3720 0.0726 -0.4386 -0.0446 -0.0260 0.2153 -0.0342 -0.1888

C27 0.1780 -0.0634 0.3807 0.0700 -0.0031 -0.1273 -0.1355 0.0385 0.0377 0.1007 0.0530

C28 0.2047 0.0032 0.0522 0.0671 -0.2049 -0.2203 0.3071 0.0820 -0.2103 -0.1211 0.0767

C29 0.0381 0.1220 -0.1941 -0.3192 -0.4769 -0.1362 -0.4529 0.0127 0.0760 0.2221 -0.0721

C30 0.1274 0.3327 -0.1960 -0.1313 -0.0532 -0.0888 -0.1002 0.3311 -0.4699 0.0323 0.1208

C31 0.1925 -0.0622 -0.0018 0.0706 0.2942 0.2312 -0.2682 0.0590 0.1415 0.3070 0.1936

APPENDIX D

PICRUSt DATA

120

PICRUSt Output Data Big Big Slag RR Creosote #OTU ID Marsh KEGG Pathways Marsh Dunes Ditch Marsh SW 1,1,1-Trichloro- Metabolism; Xenobiotics 2,2-bis(4- Biodegradation and Metabolism; chlorophenyl)etha 883 78 4369 3291 0 1,1,1-Trichloro-2,2-bis(4- ne (DDT) chlorophenyl)ethane (DDT) degradation degradation Environmental Information ABC transporters 2352283 2002231 3308320 3573946 2395820 Processing; Membrane Transport; ABC transporters

Cellular Processes; Cell Adherens junction 0 0 0 0 0 Communication; Adherens junction

Organismal Systems; Endocrine Adipocytokine 72248 57816 122742 130465 71146 System; Adipocytokine signaling signaling pathway pathway

African Human Diseases; Infectious 14066 3269 7703 15393 9836 trypanosomiasis Diseases; African trypanosomiasis

Alanine, aspartate Metabolism; Amino Acid and glutamate 519562 399347 731320 836422 581867 Metabolism; Alanine, aspartate and metabolism glutamate metabolism

Aldosterone- Organismal Systems; Excretory regulated sodium 0 0 0 0 0 System; Aldosterone-regulated reabsorption sodium reabsorption

Human Diseases; Alzheimer's 64726 49543 66318 50308 95124 Neurodegenerative Diseases; disease Alzheimer's disease Amino acid Unclassified; Metabolism; Amino 113707 83468 149642 157230 107327 metabolism acid metabolism Metabolism; Amino Acid Amino acid 676023 530281 959262 1060595 781155 Metabolism; Amino acid related related enzymes enzymes Amino sugar and Metabolism; Carbohydrate nucleotide sugar 485436 297229 748263 949083 435991 Metabolism; Amino sugar and metabolism nucleotide sugar metabolism Genetic Information Processing; Aminoacyl-tRNA 535676 477863 760057 859795 586330 Translation; Aminoacyl-tRNA biosynthesis biosynthesis

Metabolism; Xenobiotics Aminobenzoate 419881 303373 550384 479102 454125 Biodegradation and Metabolism; degradation Aminobenzoate degradation

Human Diseases; Infectious Amoebiasis 8308 7861 15823 21070 19050 Diseases; Amoebiasis Human Diseases; Amyotrophic Neurodegenerative Diseases; lateral sclerosis 51017 32403 36227 35509 60103 Amyotrophic lateral sclerosis (ALS) (ALS) Antigen Organismal Systems; Immune processing and 10489 13496 19834 17422 18102 System; Antigen processing and presentation presentation 121

Cellular Processes; Cell Growth Apoptosis 18075 15168 16142 6381 23196 and Death; Apoptosis

Arachidonic acid Metabolism; Lipid Metabolism; 43384 33590 68408 63508 76676 metabolism Arachidonic acid metabolism

Arginine and Metabolism; Amino Acid proline 722340 514337 1002143 1108128 763292 Metabolism; Arginine and proline metabolism metabolism

Arrhythmogenic Human Diseases; Cardiovascular right ventricular Diseases; Arrhythmogenic right 0 0 0 0 0 cardiomyopathy ventricular cardiomyopathy (ARVC) (ARVC)

Ascorbate and Metabolism; Carbohydrate aldarate 131244 91879 180418 189581 151981 Metabolism; Ascorbate and metabolism aldarate metabolism

Metabolism; Xenobiotics Atrazine 46256 29731 47073 31907 40140 Biodegradation and Metabolism; degradation Atrazine degradation

Bacterial Cellular Processes; Cell Motility; 396949 426981 326478 219036 590800 chemotaxis Bacterial chemotaxis Human Diseases; Infectious Bacterial invasion 626 2 272 19 0 Diseases; Bacterial invasion of of epithelial cells epithelial cells

Bacterial motility Cellular Processes; Cell Motility; 1108981 1260978 985003 614314 1881563 proteins Bacterial motility proteins

Environmental Information Bacterial secretion 469834 369312 554126 512475 585485 Processing; Membrane Transport; system Bacterial secretion system

Environmental Information Bacterial toxins 58149 50804 80400 93150 83836 Processing; Signaling Molecules and Interaction; Bacterial toxins

Genetic Information Processing; Basal transcription 6195 622 13202 22594 2 Transcription; Basal transcription factors factors Genetic Information Processing; Base excision 289813 194438 444607 558093 287934 Replication and Repair; Base repair excision repair

Metabolism; Xenobiotics Benzoate 557081 483520 738170 618058 589584 Biodegradation and Metabolism; degradation Benzoate degradation

Metabolism; Biosynthesis of Other Betalain 859 160 2298 996 0 Secondary Metabolites; Betalain biosynthesis biosynthesis Organismal Systems; Digestive Bile secretion 4989 1126 7641 8949 8 System; Bile secretion Biosynthesis and Unclassified; Metabolism; biodegradation of 54945 37186 62971 54856 48233 Biosynthesis and biodegradation of secondary secondary metabolites metabolites Biosynthesis of Metabolism; Metabolism of 12-, 14- and 16- Terpenoids and Polyketides; 239 109 0 0 0 membered Biosynthesis of 12-, 14- and 16- macrolides membered macrolides 122

Metabolism; Metabolism of Biosynthesis of 31213 28777 45318 48915 30387 Terpenoids and Polyketides; ansamycins Biosynthesis of ansamycins Biosynthesis of Metabolism; Metabolism of siderophore group Terpenoids and Polyketides; 39941 16874 40848 46048 38648 nonribosomal Biosynthesis of siderophore group peptides nonribosomal peptides Metabolism; Metabolism of Biosynthesis of Terpenoids and Polyketides; type II polyketide 0 0 0 0 0 Biosynthesis of type II polyketide backbone backbone Metabolism; Metabolism of Biosynthesis of Terpenoids and Polyketides; type II polyketide 504 100 0 0 0 Biosynthesis of type II polyketide products products Biosynthesis of Metabolism; Lipid Metabolism; unsaturated fatty 186689 125125 261952 260089 227786 Biosynthesis of unsaturated fatty acids acids Metabolism; Metabolism of Biosynthesis of Terpenoids and Polyketides; vancomycin group 27320 23639 31699 35134 38651 Biosynthesis of vancomycin group antibiotics antibiotics Metabolism; Metabolism of Biotin metabolism 86513 70364 113369 101362 103698 Cofactors and Vitamins; Biotin metabolism

Metabolism; Xenobiotics Bisphenol 98342 43879 169194 168983 53722 Biodegradation and Metabolism; degradation Bisphenol degradation

Human Diseases; Cancers; Bladder Bladder cancer 9517 6899 19603 26379 19239 cancer

Butanoate Metabolism; Carbohydrate 821708 672070 1168734 1121678 897206 metabolism Metabolism; Butanoate metabolism

Butirosin and Metabolism; Biosynthesis of Other neomycin 19871 10113 50864 73441 20431 Secondary Metabolites; Butirosin biosynthesis and neomycin biosynthesis

C5-Branched Metabolism; Carbohydrate dibasic acid 168599 151261 257226 279885 186564 Metabolism; C5-Branched dibasic metabolism acid metabolism

Environmental Information CAM ligands 0 0 0 0 0 Processing; Signaling Molecules and Interaction; CAM ligands

Metabolism; Biosynthesis of Other Caffeine 2887 252 2 0 0 Secondary Metabolites; Caffeine metabolism metabolism

Environmental Information Calcium signaling 37 0 308 32 0 Processing; Signal Transduction; pathway Calcium signaling pathway

Metabolism; Xenobiotics Caprolactam 247662 180168 330528 289536 255263 Biodegradation and Metabolism; degradation Caprolactam degradation

Carbohydrate Organismal Systems; Digestive digestion and 3817 198 502 281 5 System; Carbohydrate digestion absorption and absorption 123

Carbohydrate Unclassified; Metabolism; 71392 19872 90774 118428 38768 metabolism Carbohydrate metabolism Carbon fixation in Metabolism; Energy Metabolism; photosynthetic 245946 192705 353237 373515 259840 Carbon fixation in photosynthetic organisms organisms Carbon fixation Metabolism; Energy Metabolism; pathways in 655311 632548 879444 871508 747138 Carbon fixation pathways in prokaryotes prokaryotes Organismal Systems; Circulatory Cardiac muscle 28577 25298 30460 17199 58836 System; Cardiac muscle contraction contraction Metabolism; Metabolism of Carotenoid 44985 17269 29038 22633 17229 Terpenoids and Polyketides; biosynthesis Carotenoid biosynthesis Cellular Processes; Cell Growth Cell cycle 0 0 0 0 0 and Death; Cell cycle

Cell cycle - Cellular Processes; Cell Growth 319251 227190 311228 321935 282152 Caulobacter and Death; Cell cycle - Caulobacter

Cellular Processes; Cell Growth Cell cycle - yeast 0 0 0 0 0 and Death; Cell cycle - yeast

Unclassified; Cellular Processes Cell division 44024 44855 56931 48030 67932 and Signaling; Cell division

Unclassified; Cellular Processes Cell motility and 118421 110741 168264 130689 217822 and Signaling; Cell motility and secretion secretion

Environmental Information Cellular antigens 49845 40634 61479 54140 94951 Processing; Signaling Molecules and Interaction; Cellular antigens

Chagas disease Human Diseases; Infectious (American 10409 2868 8007 14576 19645 Diseases; Chagas disease trypanosomiasis) (American trypanosomiasis)

Genetic Information Processing; Chaperones and 509428 452699 723831 713024 776947 Folding, Sorting and Degradation; folding catalysts Chaperones and folding catalysts

Metabolism; Xenobiotics Chloroalkane and Biodegradation and Metabolism; chloroalkene 194710 145344 278034 270532 213120 Chloroalkane and chloroalkene degradation degradation

Chlorocyclohexan Metabolism; Xenobiotics e and Biodegradation and Metabolism; 80199 52942 98482 79666 93835 chlorobenzene Chlorocyclohexane and degradation chlorobenzene degradation

Cholinergic Organismal Systems; Nervous 0 0 0 0 0 synapse System; Cholinergic synapse Genetic Information Processing; Chromosome 710546 689397 953402 949036 1015910 Replication and Repair; Chromosome Chronic myeloid Human Diseases; Cancers; Chronic 0 0 0 68 0 leukemia myeloid leukemia Organismal Systems; Circadian rhythm - 8351 1311 14954 25193 6 Environmental Adaptation; plant Circadian rhythm - plant 124

Metabolism; Carbohydrate Citrate cycle 491829 378980 750875 854953 521376 Metabolism; Citrate cycle (TCA (TCA cycle) cycle) Metabolism; Biosynthesis of Other Clavulanic acid 186 14 2 0 0 Secondary Metabolites; Clavulanic biosynthesis acid biosynthesis Human Diseases; Cancers; Colorectal cancer 13754 8753 6235 1669 14723 Colorectal cancer Complement and Organismal Systems; Immune coagulation 0 0 0 0 0 System; Complement and cascades coagulation cascades Metabolism; Metabolism of Other Cyanoamino acid 145183 78305 175193 199953 146292 Amino Acids; Cyanoamino acid metabolism metabolism Cysteine and Metabolism; Amino Acid methionine 470497 406824 581679 654931 600045 Metabolism; Cysteine and metabolism methionine metabolism Metabolism; Enzyme Families; Cytochrome P450 1633 211 0 0 0 Cytochrome P450

Environmental Information Cytokine receptors 0 0 0 0 0 Processing; Signaling Molecules and Interaction; Cytokine receptors

Environmental Information Cytokine-cytokine Processing; Signaling Molecules receptor 0 0 0 0 0 and Interaction; Cytokine-cytokine interaction receptor interaction

Cytoskeleton Cellular Processes; Cell Motility; 120273 111831 159240 158384 142923 proteins Cytoskeleton proteins

Organismal Systems; Immune Cytosolic DNA- 0 0 0 0 0 System; Cytosolic DNA-sensing sensing pathway pathway Metabolism; Metabolism of Other D-Alanine 49536 38472 79015 102803 40627 Amino Acids; D-Alanine metabolism metabolism

D-Arginine and D- Metabolism; Metabolism of Other ornithine 6574 1608 9009 6294 1359 Amino Acids; D-Arginine and D- metabolism ornithine metabolism

D-Glutamine and Metabolism; Metabolism of Other D-glutamate 76667 59451 88251 99556 67129 Amino Acids; D-Glutamine and D- metabolism glutamate metabolism

DNA repair and Genetic Information Processing; recombination 1423949 1154286 2000604 2296050 1646932 Replication and Repair; DNA proteins repair and recombination proteins

Genetic Information Processing; DNA replication 278734 233230 405620 475785 313066 Replication and Repair; DNA replication

Genetic Information Processing; DNA replication 474955 400618 658110 738751 557230 Replication and Repair; DNA proteins replication proteins 125

Dilated Human Diseases; Cardiovascular cardiomyopathy 0 0 0 0 0 Diseases; Dilated cardiomyopathy (DCM) (DCM)

Metabolism; Xenobiotics Dioxin 60720 55336 71306 52892 54945 Biodegradation and Metabolism; degradation Dioxin degradation

Metabolism; Xenobiotics Drug metabolism - Biodegradation and Metabolism; 165311 117956 183423 149357 220251 cytochrome P450 Drug metabolism - cytochrome P450

Metabolism; Xenobiotics Drug metabolism - 141089 87235 191690 251297 129737 Biodegradation and Metabolism; other enzymes Drug metabolism - other enzymes

Environmental Information ECM-receptor Processing; Signaling Molecules 0 0 0 0 0 interaction and Interaction; ECM-receptor interaction Unclassified; Cellular Processes Electron transfer 15791 7384 26567 33356 3662 and Signaling; Electron transfer carriers carriers Endocrine and Organismal Systems; Excretory other factor- System; Endocrine and other 0 0 0 0 0 regulated calcium factor-regulated calcium reabsorption reabsorption Cellular Processes; Transport and Endocytosis 114 35 0 0 0 Catabolism; Endocytosis Energy Unclassified; Metabolism; Energy 489697 463231 712025 611834 606837 metabolism metabolism Epithelial cell Human Diseases; Infectious signaling in 21690 32012 33449 23174 27698 Diseases; Epithelial cell signaling Helicobacter in Helicobacter pylori infection pylori infection

Environmental Information ErbB signaling 0 0 0 0 0 Processing; Signal Transduction; pathway ErbB signaling pathway

Ether lipid Metabolism; Lipid Metabolism; 12702 7540 24219 21404 18098 metabolism Ether lipid metabolism

Metabolism; Xenobiotics Ethylbenzene 51746 33273 83063 80747 48473 Biodegradation and Metabolism; degradation Ethylbenzene degradation

Organismal Systems; Digestive Fat digestion and 0 0 0 0 0 System; Fat digestion and absorption absorption

Fatty acid Metabolism; Lipid Metabolism; 328483 269557 438471 418755 393768 biosynthesis Fatty acid biosynthesis

Fatty acid Metabolism; Lipid Metabolism; elongation in 0 0 0 0 0 Fatty acid elongation in mitochondria mitochondria

Fatty acid Metabolism; Lipid Metabolism; 679838 522669 930465 849212 694921 metabolism Fatty acid metabolism 126

Organismal Systems; Immune Fc epsilon RI 0 0 0 0 0 System; Fc epsilon RI signaling signaling pathway pathway Fc gamma R- Organismal Systems; Immune mediated 114 35 0 0 0 System; Fc gamma R-mediated phagocytosis phagocytosis Cellular Processes; Cell Motility; Flagellar assembly 472011 483186 297332 163575 678598 Flagellar assembly

Flavone and Metabolism; Biosynthesis of Other flavonol 646 17 1726 2614 3 Secondary Metabolites; Flavone biosynthesis and flavonol biosynthesis

Metabolism; Biosynthesis of Other Flavonoid 10584 1590 25253 36407 372 Secondary Metabolites; Flavonoid biosynthesis biosynthesis

Metabolism; Xenobiotics Fluorobenzoate 44299 27652 54386 43528 37228 Biodegradation and Metabolism; degradation Fluorobenzoate degradation

Cellular Processes; Cell Focal adhesion 0 0 0 0 0 Communication; Focal adhesion

Metabolism; Metabolism of Folate 262613 197678 325005 350303 306971 Cofactors and Vitamins; Folate biosynthesis biosynthesis Fructose and Metabolism; Carbohydrate mannose 302777 163153 431123 553726 229376 Metabolism; Fructose and mannose metabolism metabolism

Unclassified; Poorly Characterized; Function unknown 1061159 827315 1187511 1186268 1240047 Function unknown

Environmental Information G protein-coupled Processing; Signaling Molecules 0 0 0 0 0 receptors and Interaction; G protein-coupled receptors Environmental Information GTP-binding Processing; Signaling Molecules 0 0 0 0 0 proteins and Interaction; GTP-binding proteins

Galactose Metabolism; Carbohydrate 195172 93796 309828 430773 134466 metabolism Metabolism; Galactose metabolism

Gastric acid Organismal Systems; Digestive 0 0 0 0 0 secretion System; Gastric acid secretion

General function Unclassified; Poorly Characterized; 2151119 1733143 2796828 2867890 2434566 prediction only General function prediction only

Metabolism; Metabolism of Geraniol 353631 281292 502653 438661 381348 Terpenoids and Polyketides; degradation Geraniol degradation

Unclassified; Cellular Processes Germination 17410 17684 1 1 12 and Signaling; Germination

Glioma 0 0 0 0 0 Human Diseases; Cancers; Glioma

Glutamatergic Organismal Systems; Nervous 42395 24696 58914 75408 33131 synapse System; Glutamatergic synapse 127

Metabolism; Metabolism of Other Glutathione 302093 228840 361790 342072 453310 Amino Acids; Glutathione metabolism metabolism Environmental Information Glycan bindng Processing; Signaling Molecules 0 0 0 0 0 proteins and Interaction; Glycan bindng proteins Glycan Unclassified; Metabolism; Glycan biosynthesis and 33076 37638 38870 20376 75077 biosynthesis and metabolism metabolism

Glycerolipid Metabolism; Lipid Metabolism; 212460 138573 270124 310879 197561 metabolism Glycerolipid metabolism

Glycerophospholi Metabolism; Lipid Metabolism; 280810 232494 394379 404417 347674 pid metabolism Glycerophospholipid metabolism

Glycine, serine Metabolism; Amino Acid and threonine 571177 417404 777817 846671 634813 Metabolism; Glycine, serine and metabolism threonine metabolism Metabolism; Carbohydrate Glycolysis / 602712 408452 905528 1062213 567701 Metabolism; Glycolysis / Gluconeogenesis Gluconeogenesis

Metabolism; Glycan Biosynthesis Glycosaminoglyca and Metabolism; n biosynthesis - 0 0 0 0 0 Glycosaminoglycan biosynthesis - chondroitin sulfate chondroitin sulfate

Metabolism; Glycan Biosynthesis Glycosaminoglyca 6559 852 21675 30375 9262 and Metabolism; n degradation Glycosaminoglycan degradation

Metabolism; Glycan Biosynthesis Glycosphingolipid and Metabolism; biosynthesis - 5589 826 18083 25285 9259 Glycosphingolipid biosynthesis - ganglio series ganglio series Metabolism; Glycan Biosynthesis Glycosphingolipid and Metabolism; biosynthesis - 17512 1329 36597 55569 18519 Glycosphingolipid biosynthesis - globo series globo series Glycosphingolipid Metabolism; Glycan Biosynthesis biosynthesis - and Metabolism; 661 5 1 0 0 lacto and neolacto Glycosphingolipid biosynthesis - series lacto and neolacto series Glycosylphosphati Metabolism; Glycan Biosynthesis dylinositol(GPI)- and Metabolism; 0 0 0 0 0 anchor Glycosylphosphatidylinositol(GPI)- biosynthesis anchor biosynthesis Metabolism; Glycan Biosynthesis Glycosyltransferas 180223 147327 211855 207878 232840 and Metabolism; es Glycosyltransferases Glyoxylate and Metabolism; Carbohydrate dicarboxylate 556179 510201 736186 663783 707293 Metabolism; Glyoxylate and metabolism dicarboxylate metabolism

GnRH signaling Organismal Systems; Endocrine 114 35 0 0 0 pathway System; GnRH signaling pathway 128

Environmental Information Hedgehog 0 0 0 0 0 Processing; Signal Transduction; signaling pathway Hedgehog signaling pathway

Hematopoietic cell Organismal Systems; Immune 1141 140 0 0 0 lineage System; Hematopoietic cell lineage

Human Diseases; Infectious Hepatitis C 0 0 0 0 0 Diseases; Hepatitis C

Histidine Metabolism; Amino Acid 347669 276701 525915 585843 378026 metabolism Metabolism; Histidine metabolism

Genetic Information Processing; Homologous 373690 322979 534844 619967 447415 Replication and Repair; recombination Homologous recombination

Human Diseases; Huntington's 81406 60042 76877 60119 129179 Neurodegenerative Diseases; disease Huntington's disease

Hypertrophic Human Diseases; Cardiovascular cardiomyopathy 1073 647 498 102 9819 Diseases; Hypertrophic (HCM) cardiomyopathy (HCM)

Metabolism; Biosynthesis of Other Indole alkaloid 957 161 120 36 0 Secondary Metabolites; Indole biosynthesis alkaloid biosynthesis Human Diseases; Infectious Influenza A 13754 8753 6235 1669 14723 Diseases; Influenza A

Inorganic ion Unclassified; Cellular Processes transport and 216239 188256 276036 293591 315796 and Signaling; Inorganic ion metabolism transport and metabolism

Metabolism; Carbohydrate Inositol phosphate 147810 70665 189363 219424 125911 Metabolism; Inositol phosphate metabolism metabolism

Insulin signaling Organismal Systems; Endocrine 30638 21824 50100 54890 29067 pathway System; Insulin signaling pathway

Environmental Information Ion channels 15047 7444 17098 25239 9773 Processing; Signaling Molecules and Interaction; Ion channels

Metabolism; Biosynthesis of Other Isoflavonoid 256 551 0 0 0 Secondary Metabolites; biosynthesis Isoflavonoid biosynthesis

Isoquinoline Metabolism; Biosynthesis of Other alkaloid 31705 23464 43048 47197 31688 Secondary Metabolites; biosynthesis Isoquinoline alkaloid biosynthesis

Human Diseases; Infectious Leishmaniasis 0 0 0 0 0 Diseases; Leishmaniasis Leukocyte Organismal Systems; Immune transendothelial 0 0 0 0 0 System; Leukocyte transendothelial migration migration

Metabolism; Metabolism of Limonene and 347749 209452 518651 464395 293996 Terpenoids and Polyketides; pinene degradation Limonene and pinene degradation 129

Linoleic acid Metabolism; Lipid Metabolism; 47757 30262 74441 81531 50116 metabolism Linoleic acid metabolism

Lipid biosynthesis Metabolism; Lipid Metabolism; 536055 419502 740857 715036 623079 proteins Lipid biosynthesis proteins

Unclassified; Metabolism; Lipid Lipid metabolism 66179 53846 111868 88736 67457 metabolism Metabolism; Metabolism of Lipoic acid 39515 24522 54265 66525 40231 Cofactors and Vitamins; Lipoic metabolism acid metabolism

Metabolism; Glycan Biosynthesis Lipopolysaccharid 147510 118660 196405 128887 245343 and Metabolism; e biosynthesis Lipopolysaccharide biosynthesis

Metabolism; Glycan Biosynthesis Lipopolysaccharid and Metabolism; e biosynthesis 185304 148991 253409 184254 320648 Lipopolysaccharide biosynthesis proteins proteins Long-term Organismal Systems; Nervous 0 0 0 0 0 depression System; Long-term depression

Long-term Organismal Systems; Nervous 0 0 0 0 0 potentiation System; Long-term potentiation

Lysine Metabolism; Amino Acid 337612 283361 474914 533465 393700 biosynthesis Metabolism; Lysine biosynthesis

Lysine Metabolism; Amino Acid 436935 324591 604768 558728 442108 degradation Metabolism; Lysine degradation

Cellular Processes; Transport and Lysosome 14006 1268 35072 51279 18542 Catabolism; Lysosome

Environmental Information MAPK signaling 0 0 0 0 0 Processing; Signal Transduction; pathway MAPK signaling pathway

Environmental Information MAPK signaling 20258 21786 22103 12856 29028 Processing; Signal Transduction; pathway - yeast MAPK signaling pathway - yeast

Human Diseases; Infectious Measles 0 0 0 0 0 Diseases; Measles

Cellular Processes; Cell Growth Meiosis - yeast 22316 11033 18418 21741 27537 and Death; Meiosis - yeast

Organismal Systems; Endocrine Melanogenesis 1 0 2178 960 0 System; Melanogenesis

Membrane and Unclassified; Cellular Processes intracellular 399348 325491 458507 335973 688273 and Signaling; Membrane and structural intracellular structural molecules molecules Metabolism of Unclassified; Metabolism; cofactors and 128044 94142 139373 118910 136764 Metabolism of cofactors and vitamins vitamins 130

Metabolism; Xenobiotics Metabolism of Biodegradation and Metabolism; xenobiotics by 160354 117530 179811 143538 219881 Metabolism of xenobiotics by cytochrome P450 cytochrome P450

Methane Metabolism; Energy Metabolism; 563289 507119 790397 815616 577881 metabolism Methane metabolism

Organismal Systems; Digestive Mineral absorption 13634 13893 17449 20912 19053 System; Mineral absorption Genetic Information Processing; Mismatch repair 338289 299229 483877 543910 390634 Replication and Repair; Mismatch repair Metabolism; Glycan Biosynthesis N-Glycan 13717 8675 18901 28770 10561 and Metabolism; N-Glycan biosynthesis biosynthesis Organismal Systems; Immune NOD-like receptor 10632 13498 19872 17426 18102 System; NOD-like receptor signaling pathway signaling pathway

Metabolism; Xenobiotics Naphthalene 213637 148119 286430 244511 199349 Biodegradation and Metabolism; degradation Naphthalene degradation

Environmental Information Neuroactive Processing; Signaling Molecules ligand-receptor 0 0 0 0 0 and Interaction; Neuroactive interaction ligand-receptor interaction

Organismal Systems; Nervous Neurotrophin 0 0 0 0 0 System; Neurotrophin signaling signaling pathway pathway

Nicotinate and Metabolism; Metabolism of nicotinamide 259686 205780 337055 394340 289478 Cofactors and Vitamins; Nicotinate metabolism and nicotinamide metabolism

Nitrogen Metabolism; Energy Metabolism; 434166 367892 593975 597898 501896 metabolism Nitrogen metabolism

Metabolism; Xenobiotics Nitrotoluene 47599 70443 66733 49321 55999 Biodegradation and Metabolism; degradation Nitrotoluene degradation

Genetic Information Processing; Non-homologous 48721 33220 78327 95309 28658 Replication and Repair; Non- end-joining homologous end-joining

Environmental Information Notch signaling 0 0 0 68 0 Processing; Signal Transduction; pathway Notch signaling pathway

Metabolism; Biosynthesis of Other Novobiocin 70637 60375 108286 123016 89629 Secondary Metabolites; biosynthesis Novobiocin biosynthesis

Genetic Information Processing; Nucleotide 187963 145593 303958 379503 174563 Replication and Repair; Nucleotide excision repair excision repair

Nucleotide Unclassified; Metabolism; 26621 32523 12483 6985 26338 metabolism Nucleotide metabolism 131

Olfactory Organismal Systems; Sensory 0 0 0 0 0 transduction System; Olfactory transduction Metabolism; Metabolism of One carbon pool 284007 243328 362507 402512 325548 Cofactors and Vitamins; One by folate carbon pool by folate

Cellular Processes; Cell Growth Oocyte meiosis 0 0 0 0 0 and Death; Oocyte meiosis

Metabolism; Glycan Biosynthesis Other glycan 29377 3256 66653 99724 28376 and Metabolism; Other glycan degradation degradation Unclassified; Cellular Processes Other ion-coupled 750552 585552 728281 782753 894142 and Signaling; Other ion-coupled transporters transporters

Unclassified; Cellular Processes Other transporters 132322 103332 157989 140708 197282 and Signaling; Other transporters

Other types of O- Metabolism; Glycan Biosynthesis glycan 0 0 0 0 0 and Metabolism; Other types of O- biosynthesis glycan biosynthesis

Others 565931 345838 769547 804405 547508 Unclassified; Metabolism; Others

Oxidative Metabolism; Energy Metabolism; 786401 618422 1098789 1130793 950814 phosphorylation Oxidative phosphorylation

PPAR signaling Organismal Systems; Endocrine 182439 140868 278451 272347 188662 pathway System; PPAR signaling pathway

Human Diseases; Cancers; Pancreatic cancer 0 0 0 0 0 Pancreatic cancer Pancreatic Organismal Systems; Digestive 0 0 0 0 0 secretion System; Pancreatic secretion

Metabolism; Metabolism of Pantothenate and 322166 277782 478174 527460 362131 Cofactors and Vitamins; CoA biosynthesis Pantothenate and CoA biosynthesis

Human Diseases; Parkinson's 42331 34051 36695 18868 73559 Neurodegenerative Diseases; disease Parkinson's disease Pathogenic Human Diseases; Infectious Escherichia coli 0 0 0 0 0 Diseases; Pathogenic Escherichia infection coli infection Pathways in Human Diseases; Cancers; 41424 31390 50936 49452 52441 cancer Pathways in cancer

Penicillin and Metabolism; Biosynthesis of Other cephalosporin 41933 32192 69914 81822 66279 Secondary Metabolites; Penicillin biosynthesis and cephalosporin biosynthesis

Pentose and Metabolism; Carbohydrate glucuronate 249936 137954 322749 401004 224802 Metabolism; Pentose and interconversions glucuronate interconversions Metabolism; Carbohydrate Pentose phosphate 356495 276097 549373 659019 399647 Metabolism; Pentose phosphate pathway pathway Metabolism; Enzyme Families; Peptidases 872880 673151 1128715 1208336 1004732 Peptidases 132

Metabolism; Glycan Biosynthesis Peptidoglycan 350463 312983 503384 563298 423415 and Metabolism; Peptidoglycan biosynthesis biosynthesis Cellular Processes; Transport and Peroxisome 213987 160710 291528 300237 251309 Catabolism; Peroxisome Human Diseases; Infectious Pertussis 28826 23850 31820 19364 45770 Diseases; Pertussis Cellular Processes; Transport and Phagosome 0 0 0 0 0 Catabolism; Phagosome Metabolism; Amino Acid Phenylalanine 265088 191443 370688 362945 293438 Metabolism; Phenylalanine metabolism metabolism Phenylalanine, Metabolism; Amino Acid tyrosine and Metabolism; Phenylalanine, 388038 361480 559123 630890 494060 tryptophan tyrosine and tryptophan biosynthesis biosynthesis Metabolism; Biosynthesis of Other Phenylpropanoid 67903 15477 92105 120297 42165 Secondary Metabolites; biosynthesis Phenylpropanoid biosynthesis

Environmental Information Phosphatidylinosit Processing; Signal Transduction; ol signaling 52915 40326 75828 83579 60196 Phosphatidylinositol signaling system system

Phosphonate and Metabolism; Metabolism of Other phosphinate 43284 36347 52859 48476 57848 Amino Acids; Phosphonate and metabolism phosphinate metabolism

Environmental Information Phosphotransferas 125517 29073 87451 143994 51902 Processing; Membrane Transport; e system (PTS) Phosphotransferase system (PTS)

Metabolism; Energy Metabolism; Photosynthesis 146634 122386 230737 245743 159025 Photosynthesis

Photosynthesis - Metabolism; Energy Metabolism; 595 4 3872 119 4 antenna proteins Photosynthesis - antenna proteins

Photosynthesis Metabolism; Energy Metabolism; 178224 135570 246758 251421 177078 proteins Photosynthesis proteins

Organismal Systems; Sensory Phototransduction 0 0 0 0 0 System; Phototransduction

Phototransduction Organismal Systems; Sensory 0 0 0 0 0 - fly System; Phototransduction - fly

Organismal Systems; Plant-pathogen 87609 87148 92812 91570 120494 Environmental Adaptation; Plant- interaction pathogen interaction

Polycyclic Metabolism; Xenobiotics aromatic Biodegradation and Metabolism; 117109 72433 186568 189709 68915 hydrocarbon Polycyclic aromatic hydrocarbon degradation degradation

Metabolism; Metabolism of Polyketide sugar 79419 69601 119102 134859 105992 Terpenoids and Polyketides; unit biosynthesis Polyketide sugar unit biosynthesis 133

Unclassified; Cellular Processes Pores ion channels 343643 327219 329801 255725 641890 and Signaling; Pores ion channels

Porphyrin and Metabolism; Metabolism of chlorophyll 584315 522282 692656 696968 623809 Cofactors and Vitamins; Porphyrin metabolism and chlorophyll metabolism

Metabolism; Metabolism of Prenyltransferases 175768 125465 277275 356203 142245 Terpenoids and Polyketides; Prenyltransferases

Primary bile acid Metabolism; Lipid Metabolism; 19435 15132 20924 15567 17474 biosynthesis Primary bile acid biosynthesis

Human Diseases; Immune System Primary 34938 16888 55084 66913 28099 Diseases; Primary immunodeficiency immunodeficiency Human Diseases; Prion diseases 11613 8633 7890 4884 18285 Neurodegenerative Diseases; Prion diseases

Progesterone- Organismal Systems; Endocrine mediated oocyte 10489 13496 19834 17422 18102 System; Progesterone-mediated maturation oocyte maturation

Metabolism; Carbohydrate Propanoate 773122 618989 1050492 988181 829756 Metabolism; Propanoate metabolism metabolism Human Diseases; Cancers; Prostate Prostate cancer 10489 13496 19834 17422 18102 cancer Genetic Information Processing; Proteasome 28807 14730 50358 53322 18102 Folding, Sorting and Degradation; Proteasome Organismal Systems; Digestive Protein digestion 7852 2029 4028 3080 9823 System; Protein digestion and and absorption absorption

Genetic Information Processing; Protein export 291014 242984 423377 510139 313890 Folding, Sorting and Degradation; Protein export

Protein folding Unclassified; Genetic Information and associated 432234 386495 575407 577138 528613 Processing; Protein folding and processing associated processing

Metabolism; Enzyme Families; Protein kinases 262750 203300 350217 347391 326157 Protein kinases

Genetic Information Processing; Protein processing Folding, Sorting and Degradation; in endoplasmic 20586 21971 60088 60108 28574 Protein processing in endoplasmic reticulum reticulum

Proximal tubule Organismal Systems; Excretory bicarbonate 31448 15929 30340 35699 17862 System; Proximal tubule reclamation bicarbonate reclamation

Metabolism; Nucleotide Purine metabolism 1110165 863189 1521145 1716113 1256461 Metabolism; Purine metabolism

Metabolism; Nucleotide Pyrimidine 743443 616200 1070830 1247564 832937 Metabolism; Pyrimidine metabolism metabolism 134

Pyruvate Metabolism; Carbohydrate 740190 593047 973172 970890 818845 metabolism Metabolism; Pyruvate metabolism

RIG-I-like Organismal Systems; Immune receptor signaling 1908 252 5158 9249 367 System; RIG-I-like receptor pathway signaling pathway

Genetic Information Processing; RNA degradation 230650 180052 321074 362717 275227 Folding, Sorting and Degradation; RNA degradation

Genetic Information Processing; RNA polymerase 70209 60422 100315 119129 78515 Transcription; RNA polymerase

Genetic Information Processing; RNA transport 25358 40169 44115 25427 54538 Translation; RNA transport

Regulation of Cellular Processes; Cell Motility; 0 0 0 0 0 actin cytoskeleton Regulation of actin cytoskeleton

Renal cell Human Diseases; Cancers; Renal 17181 9141 24867 30293 19616 carcinoma cell carcinoma

Renin-angiotensin Organismal Systems; Endocrine 3446 859 682 979 9821 system System; Renin-angiotensin system

Replication, Unclassified; Genetic Information recombination and 355870 353972 448668 437024 495195 Processing; Replication, repair proteins recombination and repair proteins

Restriction Unclassified; Genetic Information 30152 39136 67779 50545 66364 enzyme Processing; Restriction enzyme

Metabolism; Metabolism of Retinol 68499 36517 89339 88823 67843 Cofactors and Vitamins; Retinol metabolism metabolism

Rheumatoid Human Diseases; Immune System 0 0 0 0 0 arthritis Diseases; Rheumatoid arthritis

Metabolism; Metabolism of Riboflavin 156775 112986 221431 258478 165925 Cofactors and Vitamins; Riboflavin metabolism metabolism

Genetic Information Processing; Ribosome 941454 823506 1375233 1619403 1059029 Translation; Ribosome

Ribosome Genetic Information Processing; 641640 538322 841381 907228 872692 Biogenesis Translation; Ribosome Biogenesis

Ribosome Genetic Information Processing; biogenesis in 29378 23883 53408 61493 46635 Translation; Ribosome biogenesis eukaryotes in eukaryotes

Organismal Systems; Digestive Salivary secretion 0 0 0 0 0 System; Salivary secretion

Secondary bile Metabolism; Lipid Metabolism; 3577 6380 2506 975 8446 acid biosynthesis Secondary bile acid biosynthesis

Environmental Information Secretion system 1126956 1100765 1276908 1033742 1660406 Processing; Membrane Transport; Secretion system 135

Metabolism; Metabolism of Other Selenocompound 234318 173049 292553 305137 253979 Amino Acids; Selenocompound metabolism metabolism Metabolism; Metabolism of Sesquiterpenoid 0 0 0 0 0 Terpenoids and Polyketides; biosynthesis Sesquiterpenoid biosynthesis Human Diseases; Infectious Shigellosis 224 0 14 0 0 Diseases; Shigellosis

Signal Unclassified; Cellular Processes transduction 286985 249091 345079 327238 377869 and Signaling; Signal transduction mechanisms mechanisms

Small cell lung Human Diseases; Cancers; Small 13754 8753 6235 1669 14723 cancer cell lung cancer

Sphingolipid Metabolism; Lipid Metabolism; 33446 9762 45179 63315 20197 metabolism Sphingolipid metabolism

Genetic Information Processing; Spliceosome 0 0 0 0 0 Transcription; Spliceosome

Unclassified; Cellular Processes Sporulation 73215 233384 19170 26460 174 and Signaling; Sporulation

Human Diseases; Infectious Staphylococcus 6531 285 1645 2229 958 Diseases; Staphylococcus aureus aureus infection infection Metabolism; Carbohydrate Starch and sucrose 307251 141102 428637 576796 250054 Metabolism; Starch and sucrose metabolism metabolism

Steroid Metabolism; Lipid Metabolism; 8535 1132 11817 10442 386 biosynthesis Steroid biosynthesis

Steroid hormone Metabolism; Lipid Metabolism; 20665 20246 18147 19066 25923 biosynthesis Steroid hormone biosynthesis

Stilbenoid, Metabolism; Biosynthesis of Other diarylheptanoid Secondary Metabolites; Stilbenoid, 30672 4001 73732 78275 2119 and gingerol diarylheptanoid and gingerol biosynthesis biosynthesis Metabolism; Biosynthesis of Other Streptomycin 149317 98893 246163 303193 149876 Secondary Metabolites; biosynthesis Streptomycin biosynthesis

Metabolism; Xenobiotics Styrene 71090 50300 89013 87204 68584 Biodegradation and Metabolism; degradation Styrene degradation

Metabolism; Energy Metabolism; Sulfur metabolism 195463 142288 266155 270814 258176 Sulfur metabolism

Genetic Information Processing; Sulfur relay 195185 134992 250531 266214 179684 Folding, Sorting and Degradation; system Sulfur relay system

Synthesis and Metabolism; Lipid Metabolism; degradation of 164029 140924 207182 176526 207603 Synthesis and degradation of ketone bodies ketone bodies Human Diseases; Immune System Systemic lupus 0 34 0 68 0 Diseases; Systemic lupus erythematosus erythematosus 136

Environmental Information TGF-beta 0 0 0 0 0 Processing; Signal Transduction; signaling pathway TGF-beta signaling pathway

Taurine and Metabolism; Metabolism of Other hypotaurine 82836 78477 115167 120115 116022 Amino Acids; Taurine and metabolism hypotaurine metabolism

Terpenoid Metabolism; Metabolism of backbone 330437 269261 473542 564742 324521 Terpenoids and Polyketides; biosynthesis Terpenoid backbone biosynthesis

Metabolism; Metabolism of Tetracycline 72340 74587 92438 69552 106591 Terpenoids and Polyketides; biosynthesis Tetracycline biosynthesis Metabolism; Metabolism of Thiamine 203877 160542 280116 300292 198684 Cofactors and Vitamins; Thiamine metabolism metabolism Cellular Processes; Cell Tight junction 0 0 0 0 0 Communication; Tight junction

Metabolism; Xenobiotics Toluene 140474 92701 175551 184589 143091 Biodegradation and Metabolism; degradation Toluene degradation

Human Diseases; Infectious Toxoplasmosis 13754 8753 6235 1669 14723 Diseases; Toxoplasmosis

Transcription Genetic Information Processing; 1008055 784720 1172577 1250206 1063940 factors Transcription; Transcription factors

Genetic Information Processing; Transcription 473570 413375 664025 722972 550159 Transcription; Transcription machinery machinery Unclassified; Genetic Information Transcription 2793 127 268 88 1 Processing; Transcription related related proteins proteins

Genetic Information Processing; Translation factors 208498 200668 311658 343710 279440 Translation; Translation factors

Translation Unclassified; Genetic Information 395818 395902 523591 502967 552208 proteins Processing; Translation proteins

Environmental Information Transporters 3712446 2905107 5293132 6021677 3599623 Processing; Membrane Transport; Transporters Tropane, Metabolism; Biosynthesis of Other piperidine and Secondary Metabolites; Tropane, 73450 53990 119365 123710 81152 pyridine alkaloid piperidine and pyridine alkaloid biosynthesis biosynthesis Metabolism; Amino Acid Tryptophan 527442 371991 727439 705157 565901 Metabolism; Tryptophan metabolism metabolism Human Diseases; Infectious Tuberculosis 75300 56575 105612 112063 82910 Diseases; Tuberculosis

Environmental Information Two-component 1439233 1305600 1631113 1390084 1979179 Processing; Signal Transduction; system Two-component system

Type I diabetes Human Diseases; Metabolic 22953 15963 36843 42521 20011 mellitus Diseases; Type I diabetes mellitus 137

Type II diabetes Human Diseases; Metabolic 20722 15372 28678 31564 20590 mellitus Diseases; Type II diabetes mellitus

Tyrosine Metabolism; Amino Acid 312245 212802 425667 434042 316205 metabolism Metabolism; Tyrosine metabolism

Ubiquinone and Metabolism; Metabolism of other terpenoid- Cofactors and Vitamins; 195718 144514 291334 314385 214349 quinone Ubiquinone and other terpenoid- biosynthesis quinone biosynthesis

Genetic Information Processing; Ubiquitin system 11683 1060 18255 21404 6 Folding, Sorting and Degradation; Ubiquitin system

Environmental Information VEGF signaling 0 0 0 0 0 Processing; Signal Transduction; pathway VEGF signaling pathway

Valine, leucine Metabolism; Amino Acid and isoleucine 415705 326362 646024 722318 469907 Metabolism; Valine, leucine and biosynthesis isoleucine biosynthesis Valine, leucine Metabolism; Amino Acid and isoleucine 820509 622527 1083161 1008593 867300 Metabolism; Valine, leucine and degradation isoleucine degradation

Various types of Metabolism; Glycan Biosynthesis N-glycan 0 0 0 0 0 and Metabolism; Various types of biosynthesis N-glycan biosynthesis

Organismal Systems; Circulatory Vascular smooth 0 0 0 0 0 System; Vascular smooth muscle muscle contraction contraction

Vasopressin- Organismal Systems; Excretory regulated water 0 0 0 0 0 System; Vasopressin-regulated reabsorption water reabsorption

Vibrio cholerae Human Diseases; Infectious 3351 6283 443 23 8446 infection Diseases; Vibrio cholerae infection

Human Diseases; Infectious Vibrio cholerae 37169 30592 42622 26459 67292 Diseases; Vibrio cholerae pathogenic cycle pathogenic cycle

Human Diseases; Cardiovascular Viral myocarditis 13754 8753 6235 1669 14723 Diseases; Viral myocarditis

Metabolism; Metabolism of Vitamin B6 105293 78549 142757 171038 116511 Cofactors and Vitamins; Vitamin metabolism B6 metabolism

Environmental Information Wnt signaling 0 0 0 68 0 Processing; Signal Transduction; pathway Wnt signaling pathway

Metabolism; Xenobiotics Xylene 42868 29479 49012 38702 28243 Biodegradation and Metabolism; degradation Xylene degradation

Metabolism; Metabolism of Zeatin 17247 15081 26026 30660 19264 Terpenoids and Polyketides; Zeatin biosynthesis biosynthesis 138 alpha-Linolenic Metabolism; Lipid Metabolism; 33018 29164 53261 66934 44861 acid metabolism alpha-Linolenic acid metabolism

Metabolism; Metabolism of Other beta-Alanine 421477 328094 583353 525761 452163 Amino Acids; beta-Alanine metabolism metabolism Metabolism; Biosynthesis of Other beta-Lactam 24402 24189 36615 39654 46670 Secondary Metabolites; beta- resistance Lactam resistance mRNA Genetic Information Processing; surveillance 272 203 0 0 0 Translation; mRNA surveillance pathway pathway

Environmental Information mTOR signaling 0 0 0 0 0 Processing; Signal Transduction; pathway mTOR signaling pathway p53 signaling Cellular Processes; Cell Growth 13754 8753 7525 2076 14723 pathway and Death; p53 signaling pathway

APPENDIX E

BIOLOG DATA

140

Big Marsh Well Groundwater Summer

Carbon Type A2 A3 A4 B1 B2 B3 B4 C1 C2 C3 C4 D1 D2 D3 D4 E1 Carbon source C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 C13 C14 C15 C16

D5.BM.1 1.94 1.93 1.82 1.40 1.98 2.05 1.75 1.32 1.96 2.21 1.91 1.68 1.94 1.59 2.04 2.03

D5.BM.2 1.85 0.57 1.27 1.21 1.66 1.46 1.36 1.95 1.61 2.00 1.96 2.05 2.01 1.74 1.87 2.10 1.23 0.82 1.00 D5.BM.3 0.62 1.27 0.57 1.31 1.02 1.19 1.26 0.64 0.72 0.96 1.04 0.61 1.26 avg 1.67 1.10 1.37 1.08 1.64 1.36 1.47 1.43 1.59 1.82 1.50 1.48 1.64 1.46 1.50 1.80 avg- avg control 0.05 -0.52 -0.26 -0.55 0.01 -0.27 -0.15 -0.20 -0.04 0.19 -0.12 -0.14 0.01 -0.17 -0.12 0.17 avg- avg control >.50 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00

Carbon Type E2 E3 E4 F1 F2 F3 F4 G1 G2 G3 G4 H1 H2 H3 H4 Carbon source C17 C18 C19 C20 C21 C22 C23 C24 C25 C26 C27 C28 C29 C30 C31 control

D5.BM.1 1.36 2.01 1.46 1.89 1.42 1.73 2.06 1.44 2.10 2.25 2.06 1.81 1.92 1.79 1.61 1.85

D5.BM.2 1.17 2.07 1.83 2.01 1.97 2.00 1.13 1.69 2.00 1.73 2.27 1.91 1.95 1.85 1.86 1.76

D5.BM.3 0.64 0.91 0.68 1.87 0.98 1.19 0.89 0.75 1.48 0.99 0.83 1.79 0.64 0.74 0.65 1.28 avg 1.06 1.66 1.32 1.92 1.46 1.64 1.36 1.29 1.86 1.65 1.72 1.84 1.50 1.46 1.37 1.63 avg- avg control -0.57 0.04 -0.31 0.30 -0.17 0.01 -0.27 -0.34 0.23 0.03 0.09 0.21 -0.12 -0.17 -0.25 avg- avg control >.50 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00

Big Marsh Surface Water Summer Carbon Type A2 A3 A4 B1 B2 B3 B4 C1 C2 C3 C4 D1 D2 D3 D4 E1 Carbon source C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 C13 C14 C15 C16

D5.BMSW.1 0.25 0.16 0.12 0.12 0.19 0.19 0.17 0.21 0.23 0.31 0.27 0.13 0.28 0.13 0.19 0.11

D5.BMSW.2 0.11 0.12 0.14 0.09 0.21 0.18 0.27 0.51 0.13 0.20 0.26 0.37 0.19 0.48 0.24 0.31 0.16 0.50 0.25 D5.BMSW.3 0.20 0.31 0.11 0.13 0.31 0.21 0.75 0.15 0.30 0.17 0.09 0.16 0.27 avg 0.17 0.26 0.17 0.14 0.24 0.16 0.19 0.34 0.19 0.42 0.23 0.27 0.21 0.23 0.20 0.23 avg- avg control -0.02 0.07 -0.02 -0.05 0.05 -0.03 0.00 0.16 0.00 0.23 0.04 0.08 0.03 0.05 0.01 0.04 avg- avg control >.50 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Carbon Type E2 E3 E4 F1 F2 F3 F4 G1 G2 G3 G4 H1 H2 H3 H4 Carbon source C17 C18 C19 C20 C21 C22 C23 C24 C25 C26 C27 C28 C29 C30 C31 control

D5.BMSW.1 0.17 0.19 0.19 0.15 0.23 0.16 0.19 0.20 0.34 0.13 0.14 0.15 0.14 0.10 0.24 0.13

D5.BMSW.2 0.14 0.28 0.16 0.20 0.23 0.14 0.23 0.16 0.13 0.10 0.14 0.14 0.10 0.16 0.19 0.20

D5.BMSW.3 0.11 0.12 0.14 0.25 0.12 0.10 0.10 0.15 0.19 0.12 0.12 0.19 0.25 0.13 0.13 0.24 avg 0.14 0.19 0.16 0.20 0.19 0.13 0.17 0.17 0.22 0.12 0.14 0.16 0.16 0.13 0.18 0.19 avg- avg control -0.05 0.01 -0.03 0.01 0.00 -0.06 -0.02 -0.02 0.03 -0.07 -0.05 -0.03 -0.03 -0.06 -0.01 avg- avg control >.50 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00

141

Creosote Marsh Surface Water Summer Carbon A2 A3 A4 B1 B2 B3 B4 C1 C2 C3 C4 D1 D2 D3 D4 E1 Type Carbon C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 C13 C14 C15 C16 source D5.CM.1 1.87 0.16 0.15 2.25 0.13 0.53 0.32 1.64 0.16 0.23 0.55 0.45 0.75 0.45 1.97 0.51

D5.CM.2 2.18 0.20 0.30 1.21 2.73 0.61 0.90 0.48 0.47 0.23 0.58 0.93 0.64 0.62 1.33 1.55

D5.CM.3 2.49 0.12 0.22 2.54 2.42 0.44 0.35 0.92 0.25 0.15 0.34 1.37 3.37 0.28 1.33 0.66 avg 2.18 0.16 0.22 2.00 1.76 0.52 0.52 1.01 0.29 0.20 0.49 0.92 1.59 0.45 1.54 0.91 avg-avg 1.99 -0.02 0.04 1.82 1.58 0.34 0.34 0.83 0.11 0.02 0.31 0.73 1.40 0.26 1.36 0.72 control avg- avg control 1.99 0.00 0.00 1.82 1.58 0.00 0.00 0.83 0.00 0.00 0.00 0.73 1.40 0.00 1.36 0.72 >.50 Carbon E2 E3 E4 F1 F2 F3 F4 G1 G2 G3 G4 H1 H2 H3 H4 Type Carbon C17 C18 C19 C20 C21 C22 C23 C24 C25 C26 C27 C28 C29 C30 C31 control source D5.CM.1 2.69 0.29 0.28 0.87 0.56 0.51 1.36 1.11 0.55 0.81 0.15 0.63 0.56 0.39 0.80 0.18

D5.CM.2 2.86 0.31 0.36 1.02 0.57 0.47 0.27 0.42 1.55 0.20 0.23 0.18 0.43 0.38 2.39 0.15

D5.CM.3 2.31 0.37 0.13 1.16 2.19 0.20 0.15 1.80 1.73 0.26 0.10 0.71 0.53 0.76 0.33 0.22 avg 2.62 0.32 0.26 1.02 1.11 0.40 0.59 1.11 1.28 0.42 0.16 0.51 0.50 0.51 1.17 0.18 avg-avg 2.43 0.14 0.07 0.83 0.92 0.21 0.41 0.93 1.09 0.24 -0.02 0.32 0.32 0.32 0.99 control avg- avg control 2.43 0.00 0.00 0.83 0.92 0.00 0.00 0.93 1.09 0.00 0.00 0.00 0.00 0.00 0.99 >.50

RR Ditch Surface Water Summer Carbon Type A2 A3 A4 B1 B2 B3 B4 C1 C2 C3 C4 D1 D2 D3 D4 E1 Carbon source C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 C13 C14 C15 C16

D5.RRD.1 2.26 1.21 1.00 1.83 2.42 2.14 1.76 1.29 0.62 1.88 1.12 2.17 2.80 2.57 0.81 1.94

D5.RRD.2 1.51 1.10 0.62 1.23 2.85 2.02 0.77 2.00 0.62 1.72 2.52 3.03 2.83 1.73 1.64 2.48 0.93 1.24 1.67 D5.RRD.3 2.26 2.85 1.18 0.62 2.36 1.22 1.90 1.59 2.47 2.85 1.50 0.75 2.25 avg 1.57 1.18 1.10 1.77 2.71 1.78 1.05 1.88 0.82 1.83 1.74 2.56 2.83 1.93 1.06 2.22 avg-avg control 1.07 0.69 0.60 1.28 2.21 1.28 0.55 1.39 0.32 1.34 1.25 2.06 2.33 1.44 0.57 1.73 avg-avg control>.50 1.07 0.69 0.60 1.28 2.21 1.28 0.55 1.39 0.00 1.34 1.25 2.06 2.33 1.44 0.57 1.73 Carbon Type E2 E3 E4 F1 F2 F3 F4 G1 G2 G3 G4 H1 H2 H3 H4 Carbon source C17 C18 C19 C20 C21 C22 C23 C24 C25 C26 C27 C28 C29 C30 C31 control

D5.RRD.1 1.71 0.59 0.59 2.36 1.46 2.16 1.57 2.52 1.25 1.57 3.30 1.15 0.51 0.37 1.40 0.55

D5.RRD.2 1.99 0.68 0.65 3.28 1.36 1.05 0.80 2.71 1.01 0.64 2.75 2.37 0.63 0.29 1.45 0.40

D5.RRD.3 2.04 0.52 0.48 2.10 1.71 1.36 1.14 2.50 0.87 0.44 2.49 2.10 0.59 0.40 1.52 0.54 avg 1.91 0.60 0.57 2.58 1.51 1.52 1.17 2.58 1.05 0.88 2.84 1.87 0.58 0.35 1.46 0.49 avg-avg control 1.42 0.10 0.08 2.09 1.02 1.03 0.67 2.08 0.55 0.39 2.35 1.38 0.08 -0.14 0.96 avg-avg control>.50 1.42 0.00 0.00 2.09 1.02 1.03 0.67 2.08 0.55 0.00 2.35 1.38 0.00 0.00 0.96 142

Big Marsh Groundwater Winter Carbon Type A2 A3 A4 B1 B2 B3 B4 C1 C2 C3 C4 D1 D2 D3 D4 E1 Carbon source C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 C13 C14 C15 C16

D5.SD.1 1.65 1.69 1.61 1.14 1.98 1.39 2.06 1.72 1.49 2.15 2.09 1.88 1.39 2.23 2.21 1.74

D5.SD.2 2.10 1.38 1.31 1.15 1.38 1.62 1.93 1.92 1.39 2.07 1.88 1.92 1.46 1.79 1.82 1.65 1.19 1.24 1.34 D5.SD.3 0.87 1.97 0.91 1.82 1.63 1.48 2.03 1.92 2.22 1.40 1.85 1.93 0.68 avg 1.64 1.44 1.42 1.05 1.78 1.31 1.94 1.76 1.45 2.09 1.96 2.01 1.42 1.96 1.99 1.36 avg- avg control -0.41 -0.62 -0.64 -1.00 -0.28 -0.75 -0.12 -0.30 -0.60 0.03 -0.09 -0.05 -0.64 -0.10 -0.07 -0.70 avg- avg control >.50 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Carbon Type E2 E3 E4 F1 F2 F3 F4 G1 G2 G3 G4 H1 H2 H3 H4 Carbon source C17 C18 C19 C20 C21 C22 C23 C24 C25 C26 C27 C28 C29 C30 C31 control

D5.SD.1 1.17 1.64 1.99 1.69 1.64 1.71 1.79 1.40 2.12 2.06 1.35 1.44 0.95 1.81 1.86 1.96

D5.SD.2 1.58 1.11 1.71 1.82 1.38 1.85 1.51 1.63 1.57 1.82 1.49 1.42 1.98 1.82 1.87 2.15

D5.SD.3 1.56 2.04 1.89 1.85 1.71 2.24 1.81 1.40 1.54 1.98 1.57 1.64 1.07 1.86 1.86 2.06 avg 1.44 1.60 1.87 1.79 1.58 1.93 1.70 1.47 1.75 1.95 1.47 1.50 1.33 1.83 1.86 2.06 avg- avg control -0.62 -0.46 -0.19 -0.27 -0.48 -0.12 -0.35 -0.58 -0.31 -0.10 -0.58 -0.55 -0.72 -0.22 -0.19 avg- avg control >.50 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00

Big Marsh Surface Water Winter Carbon Type A2 A3 A4 B1 B2 B3 B4 C1 C2 C3 C4 D1 D2 D3 D4 E1 Carbon source C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 C13 C14 C15 C16

D5.SD.1 0.41 0.77 0.24 0.19 0.26 0.47 0.52 0.83 0.34 0.52 0.21 0.94 0.53 0.16 0.28 0.21

D5.SD.2 0.39 0.48 0.71 0.23 0.22 1.01 0.52 0.29 0.46 0.21 0.42 0.48 0.60 0.25 1.23 0.30 0.60 1.01 0.42 D5.SD.3 0.23 0.31 0.63 0.57 0.33 0.87 0.24 0.15 0.66 0.49 0.23 0.80 0.40 avg 0.46 0.75 0.46 0.22 0.26 0.70 0.53 0.48 0.55 0.32 0.26 0.70 0.54 0.21 0.77 0.30 avg- avg control 0.10 0.39 0.10 -0.14 -0.10 0.34 0.17 0.12 0.20 -0.04 -0.10 0.34 0.18 -0.15 0.41 -0.06 avg- avg control >.50 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Carbon Type E2 E3 E4 F1 F2 F3 F4 G1 G2 G3 G4 H1 H2 H3 H4 Carbon source C17 C18 C19 C20 C21 C22 C23 C24 C25 C26 C27 C28 C29 C30 C31 control

D5.SD.1 0.77 0.28 0.27 0.21 0.48 0.47 0.46 0.60 0.49 0.18 0.39 0.32 0.47 0.47 0.13 0.13

D5.SD.2 0.61 0.59 0.40 0.21 0.19 0.35 0.37 0.15 0.15 0.21 0.17 0.25 0.22 0.71 0.17 0.52

D5.SD.3 0.42 0.44 0.39 0.38 0.28 0.19 0.14 0.19 0.21 0.16 0.14 0.26 0.23 0.19 0.11 0.42 avg 0.60 0.44 0.35 0.26 0.32 0.34 0.33 0.31 0.28 0.18 0.23 0.27 0.31 0.45 0.14 0.36 avg- avg control 0.24 0.08 -0.01 -0.10 -0.04 -0.02 -0.03 -0.05 -0.08 -0.18 -0.13 -0.09 -0.05 0.10 -0.22 avg- avg control >.50 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00

143

Creosote Marsh Sediment Winter Carbon Type A2 A3 A4 B1 B2 B3 B4 C1 C2 C3 C4 D1 D2 D3 D4 E1 Carbon source C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 C13 C14 C15 C16

D5.SD.1 0.63 1.15 1.37 1.13 0.19 1.80 1.95 0.14 0.66 0.19 0.40 2.03 2.83 1.85 0.58 1.41

D5.SD.2 2.67 1.36 1.85 1.41 1.45 1.50 1.86 1.55 0.18 0.19 0.32 2.58 2.96 1.04 2.16 0.36

D5.SD.3 0.14 1.06 0.93 1.06 0.82 0.78 2.69 1.85 0.40 0.15 0.25 1.07 2.30 1.11 1.87 0.43 avg 1.14 1.19 1.38 1.20 0.82 1.36 2.17 1.18 0.42 0.17 0.32 1.89 2.70 1.33 1.54 0.73 avg- avg control 0.86 0.91 1.10 0.92 0.53 1.08 1.88 0.90 0.13 -0.11 0.04 1.61 2.42 1.05 1.26 0.45 avg- avg control >.50 0.86 0.91 1.10 0.92 0.53 1.08 1.88 0.90 0.00 0.00 0.00 1.61 2.42 1.05 1.26 0.00 Carbon Type E2 E3 E4 F1 F2 F3 F4 G1 G2 G3 G4 H1 H2 H3 H4 Carbon source C17 C18 C19 C20 C21 C22 C23 C24 C25 C26 C27 C28 C29 C30 C31 control

D5.SD.1 2.10 0.35 0.49 2.56 1.78 1.51 0.54 0.56 0.40 0.19 0.23 0.68 0.18 0.20 0.76 0.41

D5.SD.2 1.73 0.38 0.25 0.42 1.61 1.32 0.47 2.43 2.04 0.20 1.45 0.25 1.10 0.85 1.60 0.28

D5.SD.3 0.76 0.17 0.27 0.38 1.24 1.25 0.32 2.06 1.16 0.15 0.13 0.28 0.93 0.84 1.01 0.16 avg 1.53 0.30 0.34 1.12 1.55 1.36 0.44 1.68 1.20 0.18 0.60 0.40 0.74 0.63 1.12 0.28 avg- avg control 1.25 0.02 0.05 0.84 1.26 1.08 0.16 1.40 0.91 -0.10 0.32 0.12 0.45 0.35 0.84 avg- avg control >.50 1.25 0.00 0.00 0.84 1.26 1.08 0.00 1.40 0.91 0.00 0.00 0.00 0.00 0.00 0.84

Creosote Marsh Surface Water Winter Carbon Type A2 A3 A4 B1 B2 B3 B4 C1 C2 C3 C4 D1 D2 D3 D4 E1 Carbon source C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 C13 C14 C15 C16

D5.SD.1 0.50 0.28 0.31 2.14 1.07 1.14 0.45 0.71 0.57 0.11 0.77 2.83 2.36 1.48 1.45 2.25

D5.SD.2 0.53 0.66 0.34 2.07 0.84 1.36 0.80 1.19 0.49 0.13 0.30 1.87 2.53 1.73 0.66 1.05 0.73 0.38 0.41 D5.SD.3 1.69 0.64 1.97 0.42 0.96 0.28 0.20 0.42 2.59 2.52 0.75 0.32 1.89 avg 0.59 0.44 0.35 1.96 0.85 1.49 0.56 0.95 0.45 0.15 0.50 2.43 2.47 1.32 0.81 1.73 avg- avg control 0.25 0.10 0.01 1.63 0.51 1.15 0.22 0.61 0.11 -0.19 0.16 2.09 2.13 0.98 0.47 1.39 avg- avg control >.50 0.00 0.00 0.00 1.63 0.51 1.15 0.00 0.61 0.00 0.00 0.00 2.09 2.13 0.98 0.00 1.39 Carbon Type E2 E3 E4 F1 F2 F3 F4 G1 G2 G3 G4 H1 H2 H3 H4 Carbon source C17 C18 C19 C20 C21 C22 C23 C24 C25 C26 C27 C28 C29 C30 C31 control

D5.SD.1 2.51 0.23 0.30 2.20 1.46 0.83 0.57 0.57 0.81 0.31 1.65 0.57 0.27 0.20 1.07 0.23

D5.SD.2 1.81 0.30 0.36 2.15 1.24 0.72 0.46 1.17 0.34 0.30 2.72 0.21 0.26 0.22 1.07 0.30

D5.SD.3 1.09 0.78 0.47 2.26 1.11 1.46 0.53 0.78 0.88 0.37 0.12 0.38 0.31 0.20 1.11 0.50 avg 1.80 0.43 0.38 2.20 1.27 1.00 0.52 0.84 0.68 0.33 1.50 0.38 0.28 0.21 1.08 0.34 avg- avg control 1.46 0.09 0.04 1.86 0.93 0.66 0.18 0.50 0.34 -0.01 1.16 0.05 -0.06 -0.13 0.74 avg- avg control >.50 1.46 0.00 0.00 1.86 0.93 0.66 0.00 0.50 0.00 0.00 1.16 0.00 0.00 0.00 0.74 144

RR Ditch Sediment Winter Carbon Type A2 A3 A4 B1 B2 B3 B4 C1 C2 C3 C4 D1 D2 D3 D4 E1 Carbon source C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 C13 C14 C15 C16

D5.SD.1 1.77 2.70 2.60 1.44 2.43 2.50 2.42 2.87 2.42 2.03 0.76 2.62 1.93 2.04 1.97 1.21

D5.SD.2 1.71 2.45 2.82 1.78 2.62 2.38 2.54 3.00 2.33 2.54 0.76 2.50 2.28 2.06 1.79 1.81 1.98 2.66 2.79 D5.SD.3 1.69 2.52 2.36 2.34 2.97 2.43 2.33 1.64 2.80 2.19 2.10 2.05 1.76 avg 1.82 2.61 2.74 1.64 2.53 2.41 2.43 2.94 2.39 2.30 1.05 2.64 2.13 2.07 1.93 1.59 avg- avg control 1.44 2.23 2.36 1.26 2.14 2.03 2.05 2.56 2.01 1.92 0.67 2.26 1.75 1.69 1.55 1.21 avg- avg control >.50 1.44 2.23 2.36 1.26 2.14 2.03 2.05 2.56 2.01 1.92 0.67 2.26 1.75 1.69 1.55 1.21 Carbon Type E2 E3 E4 F1 F2 F3 F4 G1 G2 G3 G4 H1 H2 H3 H4 Carbon source C17 C18 C19 C20 C21 C22 C23 C24 C25 C26 C27 C28 C29 C30 C31 control

D5.SD.1 1.60 0.43 1.43 1.68 2.32 2.50 1.77 2.03 1.61 0.49 2.34 1.98 0.80 2.05 2.19 0.39

D5.SD.2 1.96 0.45 0.91 1.62 1.94 2.62 1.88 2.10 1.59 0.49 2.36 1.98 0.75 2.30 1.89 0.37

D5.SD.3 1.89 0.31 0.79 1.72 2.26 2.62 1.10 2.16 1.66 0.45 2.50 2.28 0.68 2.20 1.23 0.38 avg 1.82 0.40 1.04 1.67 2.18 2.58 1.59 2.10 1.62 0.48 2.40 2.08 0.75 2.18 1.77 0.38 avg- avg control 1.44 0.02 0.66 1.29 1.80 2.20 1.21 1.71 1.24 0.10 2.02 1.70 0.37 1.80 1.39 avg- avg control >.50 1.44 0.00 0.66 1.29 1.80 2.20 1.21 1.71 1.24 0.00 2.02 1.70 0.00 1.80 1.39

RR Ditch Surface Water Winter Carbon Type A2 A3 A4 B1 B2 B3 B4 C1 C2 C3 C4 D1 D2 D3 D4 E1 Carbon source C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 C13 C14 C15 C16

D5.SD.1 2.70 1.53 2.84 2.21 2.16 2.22 2.78 2.60 1.76 0.20 1.29 2.86 2.53 2.69 2.71 2.24

D5.SD.2 2.45 1.67 2.96 2.31 2.70 1.77 2.59 2.91 1.09 0.16 1.61 3.16 2.51 2.75 2.88 2.35 2.57 0.45 2.78 D5.SD.3 2.44 2.45 1.79 2.72 2.82 2.55 0.19 1.07 3.12 2.97 1.96 1.81 2.46

Avg 2.57 1.21 2.86 2.32 2.44 1.92 2.70 2.77 1.80 0.18 1.32 3.05 2.67 2.47 2.47 2.35 avg- avg control 2.27 0.91 2.56 2.02 2.13 1.62 2.39 2.47 1.50 -0.12 1.02 2.74 2.37 2.16 2.16 2.05 avg- avg control >.50 2.27 0.91 2.56 2.02 2.13 1.62 2.39 2.47 1.50 0.00 1.02 2.74 2.37 2.16 2.16 2.05 Carbon Type E2 E3 E4 F1 F2 F3 F4 G1 G2 G3 G4 H1 H2 H3 H4 Carbon source C17 C18 C19 C20 C21 C22 C23 C24 C25 C26 C27 C28 C29 C30 C31 Control

D5.SD.1 2.44 0.51 0.91 1.60 2.08 1.62 2.14 2.57 1.95 1.66 2.74 2.61 0.67 1.74 1.84 0.30

D5.SD.2 3.11 0.67 0.94 2.61 1.68 0.42 1.63 2.41 2.03 0.36 2.80 2.38 0.28 1.47 2.09 0.24

D5.SD.3 2.66 0.41 0.73 2.36 2.04 1.06 1.84 2.67 2.08 1.40 1.73 2.90 0.54 0.76 1.39 0.38

Avg 2.73 0.53 0.86 2.19 1.93 1.03 1.87 2.55 2.02 1.14 2.42 2.63 0.50 1.33 1.77 0.30 avg- avg control 2.43 0.23 0.56 1.89 1.63 0.73 1.57 2.25 1.72 0.84 2.12 2.33 0.19 1.02 1.47 avg- avg control >.50 2.43 0.00 0.56 1.89 1.63 0.73 1.57 2.25 1.72 0.84 2.12 2.33 0.00 1.02 1.47 145

Slag Dune Well #2 Groundwater Winter

A2 A3 A4 B1 B2 B3 B4 C1 C2 C3 C4 D1 D2 D3 D4 E1 Carbon source C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 C13 C14 C15 C16

D5.SD.1 0.10 0.10 0.10 0.12 0.11 0.12 0.10 0.17 0.11 0.10 0.10 0.11 0.10 0.10 0.10 0.10

D5.SD.2 0.10 0.11 0.10 0.10 0.10 0.11 0.12 0.14 0.10 0.10 0.10 0.17 0.10 0.10 0.10 0.10 0.10 0.11 0.10 D5.SD.3 0.10 0.11 0.10 0.10 0.16 0.20 0.10 0.10 0.11 0.10 0.10 0.11 0.10 avg 0.10 0.11 0.10 0.11 0.10 0.11 0.11 0.16 0.13 0.10 0.10 0.13 0.10 0.10 0.10 0.10 avg- avg control -0.01 -0.01 -0.02 -0.01 -0.01 -0.01 -0.01 0.04 0.02 -0.01 -0.02 0.02 -0.01 -0.01 -0.01 -0.01 avg- avg control >.50 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Carbon Type E2 E3 E4 F1 F2 F3 F4 G1 G2 G3 G4 H1 H2 H3 H4 Carbon source C17 C18 C19 C20 C21 C22 C23 C24 C25 C26 C27 C28 C29 C30 C31 control

D5.SD.1 0.10 0.11 0.11 0.12 0.12 0.11 0.11 0.11 0.11 0.11 0.12 0.11 0.11 0.10 0.10 0.12

D5.SD.2 0.17 0.13 0.11 0.11 0.10 0.12 0.10 0.10 1.42 0.11 0.11 0.11 0.10 0.26 0.10 0.10

D5.SD.3 0.11 0.12 0.10 0.17 0.10 0.10 0.12 0.10 0.10 0.10 0.11 0.10 0.10 0.10 0.11 0.12 avg 0.13 0.12 0.10 0.13 0.11 0.11 0.11 0.10 0.54 0.11 0.11 0.11 0.10 0.15 0.10 0.11 avg- avg control 0.01 0.01 -0.01 0.02 -0.01 -0.01 -0.01 -0.01 0.43 -0.01 0.00 -0.01 -0.01 0.04 -0.01 avg- avg control >.50 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00

Slag Dune Well #2 Groundwater Sediment Carbon Type A2 A3 A4 B1 B2 B3 B4 C1 C2 C3 C4 D1 D2 D3 D4 E1 Carbon source C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 C13 C14 C15 C16

D5.SD.1 1.79 2.50 2.35 1.15 2.08 1.00 1.19 1.87 1.45 1.36 1.65 1.54 1.76 1.31 1.57 1.51

D5.SD.2 1.87 2.56 1.75 1.11 2.03 1.18 2.17 2.21 1.15 2.30 1.49 1.82 1.08 0.96 1.74 1.15 1.83 1.07 1.39 D5.SD.3 0.70 2.00 0.78 1.38 1.87 1.33 1.35 1.17 1.53 1.43 1.28 1.86 0.65 avg 1.83 2.04 1.83 0.98 2.04 0.99 1.58 1.98 1.31 1.67 1.44 1.63 1.42 1.18 1.72 1.10 avg- avg control 0.07 0.28 0.07 -0.78 0.27 -0.78 -0.19 0.22 -0.46 -0.10 -0.33 -0.14 -0.34 -0.58 -0.04 -0.66 avg- avg control >.50 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Carbon Type E2 E3 E4 F1 F2 F3 F4 G1 G2 G3 G4 H1 H2 H3 H4 Carbon source C17 C18 C19 C20 C21 C22 C23 C24 C25 C26 C27 C28 C29 C30 C31 Control

D5.SD.1 1.59 1.33 0.78 1.13 1.83 1.05 1.07 1.29 1.85 1.72 1.93 0.98 0.93 1.23 1.21 1.60

D5.SD.2 1.35 1.96 1.44 1.39 1.03 1.39 1.21 0.83 1.31 2.08 1.78 0.67 1.08 1.01 1.24 2.13

D5.SD.3 2.15 2.35 1.36 0.58 1.87 0.55 0.98 0.86 0.57 1.34 0.55 1.69 1.05 1.73 1.35 1.57 avg 1.70 1.88 1.19 1.03 1.58 1.00 1.09 0.99 1.24 1.71 1.42 1.11 1.02 1.32 1.27 1.76 avg- avg control -0.07 0.11 -0.57 -0.73 -0.19 -0.77 -0.68 -0.77 -0.52 -0.05 -0.35 -0.66 -0.75 -0.44 -0.50 avg- avg control >.50 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00

146

Slag Dune Well #3 Groundwater Winter Carbon Type A2 A3 A4 B1 B2 B3 B4 C1 C2 C3 C4 D1 D2 D3 D4 E1 Carbon source C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 C13 C14 C15 C16 D5.SD.1 1.34 0.83 0.52 0.66 1.36 0.62 0.85 1.51 1.26 1.62 1.16 1.33 0.99 0.98 1.53 0.60 D5.SD.2 0.61 0.67 1.53 0.23 0.97 0.95 1.88 1.24 0.86 1.54 1.04 1.72 0.82 0.81 1.78 0.66 D5.SD.3 1.28 0.56 0.53 0.36 1.65 0.94 1.24 1.87 0.38 1.20 0.24 1.41 1.09 1.00 1.76 0.44 avg 1.08 0.68 0.86 0.42 1.33 0.84 1.32 1.54 0.83 1.45 0.81 1.48 0.97 0.93 1.69 0.57 avg- avg control 0.14 -0.26 -0.08 -0.53 0.39 -0.10 0.38 0.60 -0.11 0.51 -0.13 0.54 0.02 -0.01 0.75 -0.37 avg- avg control >.50 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.60 0.00 0.51 0.00 0.54 0.00 0.00 0.75 0.00 Carbon Type E2 E3 E4 F1 F2 F3 F4 G1 G2 G3 G4 H1 H2 H3 H4 Carbon source C17 C18 C19 C20 C21 C22 C23 C24 C25 C26 C27 C28 C29 C30 C31 control D5.SD.1 1.45 1.23 1.09 1.22 1.41 0.54 1.11 1.28 1.53 1.34 1.83 0.60 0.84 1.41 1.31 0.75 D5.SD.2 1.73 1.91 1.66 0.79 1.34 1.15 1.57 0.89 0.88 1.43 1.59 0.77 1.61 1.28 0.86 1.30 D5.SD.3 1.82 1.66 0.13 0.77 0.88 1.41 1.64 0.61 1.77 1.35 0.91 0.53 0.42 1.21 1.25 0.78 avg 1.67 1.60 0.96 0.93 1.21 1.03 1.44 0.93 1.40 1.37 1.45 0.63 0.96 1.30 1.14 0.94 avg- avg control 0.73 0.66 0.02 -0.01 0.27 0.09 0.50 -0.02 0.45 0.43 0.50 -0.31 0.01 0.36 0.20 avg- avg control >.50 0.73 0.66 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.50 0.00 0.00 0.00 0.00

Slag Dune Surface Water Sediment Winter Carbon Type A2 A3 A4 B1 B2 B3 B4 C1 C2 C3 C4 D1 D2 D3 D4 E1 Carbon source C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 C13 C14 C15 C16

D5.SD.1 0.09 0.09 0.09 0.11 0.10 0.09 0.09 0.22 0.09 0.09 0.09 0.10 0.09 0.09 0.09 0.09

D5.SD.2 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.19 0.09 0.09 0.11 0.09 0.08 0.09 0.08 0.09 0.09 0.09 0.09 D5.SD.3 0.10 0.09 0.09 0.09 0.16 0.08 0.09 0.09 0.09 0.09 0.09 0.09 0.09 a v g 0.09 0.09 0.09 0.10 0.10 0.09 0.09 0.19 0.09 0.09 0.10 0.10 0.09 0.09 0.09 0.09 avg- avg control 0.00 0.01 0.00 0.01 0.01 0.00 0.00 0.10 0.00 0.00 0.01 0.01 0.00 0.00 0.00 0.00 avg- avg control >.50 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Carbon Type E2 E3 E4 F1 F2 F3 F4 G1 G2 G3 G4 H1 H2 H3 H4 Carbon source C17 C18 C19 C20 C21 C22 C23 C24 C25 C26 C27 C28 C29 C30 C31 control

D5.SD.1 0.09 0.09 0.09 0.11 0.10 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09

D5.SD.2 0.09 0.09 0.09 0.10 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09

D5.SD.3 0.08 0.09 0.09 0.10 0.08 0.09 0.09 0.08 0.08 0.09 0.09 0.09 0.08 0.09 0.09 0.09 avg 0.09 0.09 0.09 0.10 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 avg- avg control 0.00 0.00 0.00 0.01 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 avg- avg control >.50 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00

147

Slag Dune Surface Water Winter Carbon Type A2 A3 A4 B1 B2 B3 B4 C1 C2 C3 C4 D1 D2 D3 D4 E1 Carbon source C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 C13 C14 C15 C16

D5.SD.1 0.36 0.11 0.11 0.10 0.12 0.10 0.12 0.12 0.11 0.11 0.14 0.22 0.36 0.10 0.10 0.11

D5.SD.2 0.10 0.11 0.10 0.10 0.10 0.58 0.89 0.18 0.10 0.10 0.10 0.19 0.10 0.15 0.10 0.10 0.11 0.11 0.10 D5.SD.3 0.16 0.11 0.10 1.44 0.21 0.33 0.10 0.10 0.18 0.10 0.10 0.11 0.12 avg 0.19 0.11 0.10 0.12 0.11 0.26 0.82 0.17 0.18 0.11 0.11 0.20 0.19 0.12 0.10 0.11 avg- avg control 0.07 -0.01 -0.02 0.00 -0.01 0.15 0.70 0.06 0.06 -0.01 0.00 0.08 0.07 0.00 -0.01 -0.01 avg- avg control >.50 0.00 0.00 0.00 0.00 0.00 0.00 0.70 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Carbon Type E2 E3 E4 F1 F2 F3 F4 G1 G2 G3 G4 H1 H2 H3 H4 Carbon source C17 C18 C19 C20 C21 C22 C23 C24 C25 C26 C27 C28 C29 C30 C31 control

D5.SD.1 0.10 0.13 0.10 0.15 0.11 0.10 0.11 0.10 0.12 0.11 0.10 0.12 0.11 0.11 0.10 0.11

D5.SD.2 2.00 0.12 0.11 0.17 0.10 0.13 0.10 0.95 0.11 0.10 0.10 0.10 0.11 0.10 0.10 0.12

D5.SD.3 0.13 0.11 0.10 0.15 0.14 0.13 0.16 1.07 0.10 0.10 0.11 0.10 0.10 0.15 0.13 0.11 avg 0.74 0.12 0.10 0.16 0.11 0.12 0.12 0.71 0.11 0.10 0.10 0.11 0.11 0.12 0.11 0.12 avg- avg control 0.63 0.00 -0.01 0.04 0.00 0.00 0.00 0.59 -0.01 -0.01 -0.01 -0.01 -0.01 0.00 -0.01 avg- avg control >.50 0.63 0.00 0.00 0.00 0.00 0.00 0.00 0.59 0.00 0.00 0.00 0.00 0.00 0.00 0.00

Wolf Lake Sediment Carbon Type A2 A3 A4 B1 B2 B3 B4 C1 C2 C3 C4 D1 D2 D3 D4 E1 Carbon source C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 C13 C14 C15 C16

D5.SD.1 2.40 1.43 1.51 1.31 1.88 1.75 2.12 1.62 0.69 0.18 0.34 2.19 1.74 1.67 1.61 1.48

D5.SD.2 0.97 0.91 1.51 1.23 2.05 1.48 1.68 2.28 1.00 0.11 0.67 2.20 1.91 1.66 1.45 1.55 1.22 1.26 1.74 D5.SD.3 0.61 0.87 1.71 1.84 1.73 0.63 0.12 0.42 1.09 1.84 1.08 0.89 1.56 avg 1.53 1.20 1.58 1.05 1.60 1.65 1.88 1.88 0.77 0.14 0.48 1.82 1.83 1.47 1.32 1.53 avg- avg control 1.31 0.98 1.37 0.83 1.38 1.43 1.66 1.66 0.55 -0.08 0.26 1.61 1.61 1.25 1.10 1.31 avg- avg control >.50 1.31 0.98 1.37 0.83 1.38 1.43 1.66 1.66 0.55 0.00 0.00 1.61 1.61 1.25 1.10 1.31 Carbon Type E2 E3 E4 F1 F2 F3 F4 G1 G2 G3 G4 H1 H2 H3 H4 Carbon source C17 C18 C19 C20 C21 C22 C23 C24 C25 C26 C27 C28 C29 C30 C31 control

D5.SD.1 1.60 0.54 1.13 1.79 1.39 2.33 1.07 2.23 1.44 0.26 0.11 1.90 0.88 2.64 0.94 0.30

D5.SD.2 1.85 0.59 0.42 1.67 2.02 1.65 1.23 2.12 2.03 0.31 1.09 1.24 0.49 2.18 1.03 0.14

D5.SD.3 1.51 0.38 0.34 1.51 1.28 1.08 0.96 2.48 1.03 0.20 1.37 0.61 1.04 1.54 0.53 0.22 avg 1.65 0.50 0.63 1.65 1.56 1.69 1.09 2.28 1.50 0.26 0.86 1.25 0.80 2.12 0.83 0.22 avg- avg control 1.44 0.28 0.41 1.43 1.34 1.47 0.87 2.06 1.28 0.04 0.64 1.03 0.58 1.90 0.62 avg- avg control >.50 1.44 0.00 0.00 1.43 1.34 1.47 0.87 2.06 1.28 0.00 0.64 1.03 0.58 1.90 0.62 148 Wolf Lake Surface Water Winter Carbon Type A2 A3 A4 B1 B2 B3 B4 C1 C2 C3 C4 D1 D2 D3 D4 E1 Carbon source C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 C13 C14 C15 C16

D5.SD.1 1.73 0.96 0.70 0.80 1.16 0.91 1.72 0.93 0.44 0.31 0.26 1.42 2.17 0.96 0.89 1.66

D5.SD.2 1.28 1.22 0.77 0.79 1.92 1.14 1.68 1.86 0.38 0.43 0.33 1.71 2.64 1.31 1.12 0.57 1.69 0.83 0.67 D5.SD.3 1.12 1.53 0.90 1.68 1.02 1.01 0.78 0.46 1.49 1.92 1.49 0.64 0.67 avg 1.57 1.00 0.71 0.90 1.54 0.98 1.69 1.27 0.61 0.50 0.35 1.54 2.24 1.25 0.88 0.97 avg- avg control 1.40 0.84 0.55 0.74 1.37 0.82 1.53 1.10 0.45 0.34 0.18 1.38 2.08 1.09 0.72 0.80 avg- avg control >.50 1.40 0.84 0.55 0.74 1.37 0.82 1.53 1.10 0.00 0.00 0.00 1.38 2.08 1.09 0.72 0.80 Carbon Type E2 E3 E4 F1 F2 F3 F4 G1 G2 G3 G4 H1 H2 H3 H4 Carbon source C17 C18 C19 C20 C21 C22 C23 C24 C25 C26 C27 C28 C29 C30 C31 control

D5.SD.1 1.96 0.26 0.32 1.27 0.94 0.51 0.28 2.33 0.92 0.52 0.16 1.32 0.32 1.43 0.62 0.17

D5.SD.2 1.88 0.25 0.31 2.17 1.38 0.58 0.33 2.24 0.83 0.24 0.14 1.64 0.31 1.24 0.52 0.15

D5.SD.3 1.79 0.17 0.27 1.33 1.14 0.68 0.50 1.51 0.60 0.25 0.12 2.31 0.32 0.92 0.52 0.18 avg 1.88 0.23 0.30 1.59 1.15 0.59 0.37 2.02 0.78 0.34 0.14 1.75 0.31 1.20 0.55 0.17 avg- avg control 1.71 0.06 0.14 1.43 0.99 0.43 0.20 1.86 0.62 0.17 -0.03 1.59 0.15 1.03 0.39 avg- avg control >.50 1.71 0.00 0.00 1.43 0.99 0.00 0.00 1.86 0.62 0.00 0.00 1.59 0.00 1.03 0.00

Big Marsh Groundwater Sediment Winter Carbon Type A2 A3 A4 B1 B2 B3 B4 C1 C2 C3 C4 D1 D2 D3 D4 E1 Carbon source C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 C13 C14 C15 C16

D5.SD.1 0.11 0.87 1.49 0.96 0.11 0.68 2.37 0.96 0.37 0.13 0.17 1.07 0.70 1.90 1.64 0.20

D5.SD.2 0.16 0.86 0.51 1.18 0.54 1.73 1.67 1.22 0.41 0.12 0.17 1.22 1.37 2.00 1.10 0.14 0.15 0.74 1.06 D5.SD.3 1.10 0.19 0.58 1.59 1.18 0.24 0.13 0.13 1.23 1.37 1.81 1.14 0.14 avg 0.14 0.82 1.02 1.08 0.28 0.99 1.88 1.12 0.34 0.12 0.16 1.17 1.15 1.90 1.29 0.16 avg- avg control 0.02 0.70 0.90 0.96 0.16 0.87 1.76 1.00 0.22 0.01 0.04 1.06 1.03 1.78 1.18 0.04 avg- avg control >.50 0.00 0.70 0.90 0.96 0.00 0.87 1.76 1.00 0.00 0.00 0.00 1.06 1.03 1.78 1.18 0.00 Carbon Type E2 E3 E4 F1 F2 F3 F4 G1 G2 G3 G4 H1 H2 H3 H4 Carbon source C17 C18 C19 C20 C21 C22 C23 C24 C25 C26 C27 C28 C29 C30 C31 control

D5.SD.1 0.76 0.18 0.12 0.16 0.92 0.19 0.17 0.24 0.16 0.12 0.14 0.21 0.35 1.81 1.09 0.10

D5.SD.2 1.59 0.23 0.17 0.20 1.48 0.54 0.21 0.27 0.15 0.14 0.14 0.30 0.20 1.94 0.59 0.15

D5.SD.3 0.79 0.14 0.11 0.21 1.13 0.41 0.13 0.26 1.00 0.11 0.10 0.20 0.19 1.60 1.17 0.11 avg 1.04 0.18 0.13 0.19 1.18 0.38 0.17 0.26 0.44 0.12 0.13 0.24 0.25 1.79 0.95 0.12 avg- avg control 0.93 0.06 0.01 0.07 1.06 0.26 0.05 0.14 0.32 0.01 0.01 0.12 0.13 1.67 0.83 avg- avg control >.50 0.93 0.00 0.00 0.00 1.06 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 1.67 0.83