The Influence of Sodium Chloride on the Performance of Gammarus Amphipods and the Community Composition of Microbes Associated with Leaf Detritus

THE INFLUENCE OF SODIUM CHLORIDE ON THE PERFORMANCE OF GAMMARUS AMPHIPODS AND THE COMMUNITY COMPOSITION OF MICROBES ASSOCIATED WITH LEAF DETRITUS

By Shelby McIlheran

Leaf litter decomposition is a fundamental part of the carbon cycle and helps support aquatic food webs along with being an important assessment of the health of rivers and streams. Disruptions in this organic matter breakdown can signal problems in other parts of ecosystems. One disruption is rising chloride concentrations.

Chloride concentrations are increasing in many rivers worldwide due to anthropogenic sources that can harm biota and affect ecosystem processes. Elevated chloride concentrations can lead to lethal or sublethal impacts. While many studies have shown that excessive chloride uptake impacts health (e.g. lowered respiration and growth rates) in a wide variety of aquatic organisms including microbes and benthic invertebrates). The impacts of high chloride concentrations on decomposers are less well understood.

My research objective was to assess how increasing chloride concentrations affect the performance and diversity of decomposer organisms in freshwater systems. I experimentally manipulated chloride concentrations in microcosms containing leaves colonized by microbes or containing leaves, microbes and amphipods. Respiration rate, decomposition, and community composition of the microbes were measured along with the amphipod growth rate, egestion rate, and mortality.

Elevated chloride concentration did not impact microbial respiration rates or leaf decomposition, but had large impacts on bacteria community composition. It did cause a decrease in instantaneous growth rate, and 100% mortality in the highest amphipod chloride treatment, but amphipod egestion rate was not significantly affected. The results of my research suggest that the widespread increases in chloride concentrations in rivers will have an impact on decomposer communities in these systems.

ACKNOWLEDGEMENTS

Support for this project came from a graduate student/faculty collaborative grant from the University of Wisconsin Office of Student Research and Creative Activity. I would like to thank the ERIC lab, Jenna Fossen, and Briana Harter for all their support in this project. Along with these individuals, I would like to thank Hannah Nauth and

Nathan Nozzi for all their help without which this project would not have happened. I would also like to thank my graduate committee members, Dr. Bob Stelzer, Dr. Sabrina

Mueller-Spitz, and Dr. Bob Pillsbury for all their help and for answering all the questions

I had.

TABLE OF CONTENTS

Page LIST OF TABLES ...... iv LIST OF FIGURES ...... v CHAPTER I – INTRODUCTION ...... 1 CHAPTER II – THE INFLUENCE OF SODIUM CHLORIDE ON THE PERFORMANCE OF GAMMARUS AMPHIPODS AND THE COMMUNITY COMPOSITION OF MICROBES ASSOCIATED WITH LEAF DETRITUS ...... 5

Introduction ...... 5 Results ...... 15 Discussion ...... 24 Acknowledgements ...... 31

CHAPTER III – CONCLUSIONS ……………………………………… ...... 32 APPENDICES

Appendix A. Genera that were present exclusively after exposure to a single chloride treatment ...... 35 Appendix B. Sequence count data for all treatments (two replicates are indicated per treatment ...... 38 Appendix C. Genera relative abundance (percentage) data for all Treatments (Two replicates are indicated per treatment) ...... 53 Appendix D. Bacteria phyla relative abundance (percentage) data for all treatments (Two replicates are indicated per treatment) ...... 68 Appendix E: Two Additional Supplemental Figures ...... 70

REFERENCES ...... 73

iii

LIST OF TABLES

Page Table 1 Specific conductance data from chloride treatment solution ...... 10

Table 2 Shannon-Weiner Diversity Index, Genera Evenness, and Richness ....19

LIST OF FIGURES Page Figure 1 Respiration rates (means+SD) from 6/6/18 after exposure to the chloride treatments ...... 15

Figure 2 Percent leaf disk mass remaining on Day 6 (6/6/18) and Day 14 (6/14/18) (means+SD) ...... 16

Figure 3 Dissolved organic carbon (DOC) (means+SD) ...... 16

Figure 4 Bacterial genera relative abundance after exposure to chloride treatments ...... 20

Figure 5 Bacteria genera sequence counts after exposure to chloride treatments ...... 21

Figure 6 Amphipod percent mortality after exposure to chloride treatments ....22

Figure 7 Amphipod instantaneous growth rates (means+SD)...... 22

Figure 8 Amphipod egestion rates (means+SD) ...... 23

Chapter I Introduction

Leaf litter decomposition in streams is necessary to break down allochthonous

(i.e. inputs from outside the system) organic matter into smaller particle sizes that can be

used as energy in ecosystems. Multiple organisms including microbes (bacteria and

fungi) and macroinvertebrates (i.e. larger invertebrates such as worms and amphipods)

break down this matter. Microbes will start decomposition by colonizing the leaf litter, which tends to make it more nutritious and available to macroinvertebrates who can consume organic matter (shredders). These shredders decrease the average particle size of the organic matter, which increases the available surface area for microbes to colonize and further facilitates decomposition. This breakdown helps carbon and energy move throughout the rest of the aquatic ecosystem (Tyree et al., 2016, Gulis et al., 2003). Water quality, including the chemical composition of streams, can affect how well this breakdown occurs. Changes in the ion concentrations of water can impact decomposition rates (Imberger et al., 2008). Chloride concentrations have been rising in many suburban and urban streams throughout the world in recent years (Gardner et al., 2010, Wallace et al., 2016, Corsi et al., 2015, Chapra et al., 2012). Chloride can enter streams, rivers, and other water bodies through anthropogenic sources such as surface water runoff from road salt, agriculture, mines, and wastewater treatment plants along with natural sources such as rock weathering, sea spray in coastal locations, and rainwater (Kaushal et al. 2018,

Cañedo-Argüelles 2016, Cañedo-Argüelles et al., 2013). A dramatic increase in road salt

application during the last several decades is a major cause for rising chloride

concentrations in streams and rivers, especially in urban areas (Kaushal et al., 2018).

Once chloride enters streams or rivers, it can have many negative effects. These

can include pH changes through variation in the number of base cations present, increases in heavy metal bioavailability and transport, and increases in lake stratification (Dugan et al. 2017, Kaushal et al. 2018). It can also cause mortality, changes in fertility and egg hatching, changes in sex ratios, decreased growth rates, and reduced larval recruitment in several animals including fish, amphibians, and invertebrates (Findlay 2011, Cañedo-

Argüelles et al., 2016, Corsi et al., 2015, Corsi et al., 2010, Lambert et al., 2016).

Chloride is able to cause these effects since it remains in solution in the water it enters instead of precipitating out like other ions. Chloride can upset the osmotic balance aquatic organisms have with the environment and cause them to expend more energy to keep this balance which can lead to negative effects such as reduced growth rates or mortality (Cañedo-Argüelles et al., 2014, Canhoto et al., 2017, Tyree et al., 2016, Findlay et al., 2011). Organisms in sub-lethal or lethal amounts (Corsi et al., 2010) can also take up chloride. How organisms are affected depends on how much chloride is in the water

(i.e. its concentration) and if it is released in chronic or short-term periods. The length of time that chloride is at high concentrations can affect its impact on organisms and ecosystem recovery (Cañedo-Argüelles et al., 2014). Along with the time, how tolerant an organism is to elevated chloride can affect how well that organism can handle the chloride concentration in the water. The taxa that are more sensitive are more likely to have decreased growth or perish. This can affect the taxa composition (e.g. biodiversity)

of the ecosystem. Reduced biodiversity may have negative effects on organic matter

decomposition rates, but there is not a consensus on the exact relationship between

biodiversity and decomposition rates since some studies show decreases in biodiversity

having little effect on the decomposition rates, while others have seen decreases in

diversity and decomposition rates (Canhoto et al., 2017, Tyree et al., 2016).

While much is known about how chloride can affect various organisms including

insects, fish, and trees (Findlay et al. 2011), there is less known on how it impacts the

process of decomposition and decomposing organisms including microbes (e.g. bacteria

and fungi) and shredders such as amphipods (a common type of macroinvertebrate found

in Wisconsin streams). The main objective of my study was to assess how increasing

chloride concentrations in streams affect the performance and community composition of

microbial decomposers and the performance of macroinvertebrate decomposers. I

predicted that as the chloride concentration increased, the microbial response would

include an increase in the percent leaf disk mass remaining (e.g. a measure of decomposition), a decrease in the microbial respiration rate, and a shift in the bacteria community composition. The predictions for the amphipod responses are that as the chloride concentration increases, the amphipod mortality will increase, the amphipod instantaneous growth rate will increase, and the amphipod egestion rate will increase. The results of my research show how chloride toxicity can impact an aquatic ecosystem process because the performance of the amphipods and the microbes may be affected.

This will affect the ecosystems and society because, as mentioned previously, chloride levels are rising in many streams and rivers in the United States and around the world and

will likely continue to rise as urbanization expands (Gardner et al., 2010, Wallace et al.,

2016, Corsi et al., 2015, Chapra et al., 2012). Knowing how chloride and road salt can impact freshwater can help communities make more informed decisions about their road salt use.

Chapter II The Influence of Sodium Chloride on the Performance of Gammarus Amphipods and the Community Composition of Microbes Associated with Leaf Detritus

Introduction Urban development and changing land use can have adverse impacts on water

quality, including salinity, in freshwater ecosystems. In recent years, attention has

focused on chloride since concentrations have been rising in many suburban and urban

streams throughout the world (Wallace et al., 2016, Corsi et al., 2015, Chapra et al., 2012,

Tyree et al., 2016, Dugan et al., 2017). Some urban streams have base flow chloride

concentration levels above 100 mg Cl-/L and up to or exceeding 7000 mg Cl-/L during

runoff events (Corsi et al., 2010). Anthropogenic sources of chloride include road salt,

fertilizer, mines, septic tank effluent, and wastewater treatment plants. Chloride can also enter water bodies through rock weathering, sea spray in coastal locations, and rainwater, but these sources are not usually considered important since they tend to be very small compared to others (Kaushal et al. 2018, Cañedo-Argüelles et al., 2016, Cañedo-

Argüelles et al., 2013, Pal et al., 2017). Increases in road salt application in the last several decades have been a major cause for the rising chloride concentrations in freshwater ecosystems, especially in urban areas; including those is the United States,

where millions of tons of road salt are used every year (Findlay et al., 2011).

Elevated chloride concentrations are known to have acute and chronic effects on

aquatic biota including increased mortality of fish and invertebrates (Findlay et al., 2011),

decreased growth rates, changes in reproduction of frogs (Lambert et al., 2016), reduced

larval recruitment and delayed development of invertebrates and amphibians (Findlay et

al.,2011, Corsi et al., 2015, Corsi et al., 2010). Most previous chloride toxicity studies on animals have been restricted to a small number of species such as fathead minnows

(Pimephales promelas) and cladocerans (Ceriodaphnia, Daphnia) (Elphick et al., 2010,

Evans et al., 2001, Corsi et al., 2010, Gardner et al., 2010). More recently, research on other groups of animals, including frogs (Lambert et al., 2016), additional species of invertebrates (Cañedo-Argüelles et al., 2012), and microbes such as fungi and bacteria, had been conducted to evaluate the lethal and sublethal effects of chloride on these taxa

(Canhoto et al., 2017, Nuy et al., 2018).

The impacts of chloride concentration on organisms depend on environmental chloride concentrations and the sensitivity of taxa to elevated salinity. Chloride is one of the most abundant ions in freshwater and it is essential for biological functions such as electrical cell potential generation, fluid regulation, and hormone signaling pathways

(Tyree et al., 2016). However, elevated chloride concentrations in freshwater can upset an organism’s osmotic balance and cause them to expend more energy for osmoregulation

(Cañedo-Argüelles et al., 2013). The species that are more sensitive to increases in chloride may not be able to maintain osmotic balance, which can lead to death. This can also lead to a decrease in how well organisms are able to perform certain functions such as growth (Cañedo-Argüelles et al., 2014, Canhoto et al., 2017, Findlay et al., 2011). The sensitivity of freshwater species to salinity stress is likely to vary among taxa, which

could cause shifts in biodiversity of freshwater ecosystems exposed to elevated chloride

concentrations. These shifts have been shown to affect ecosystem processes in various

studies where elevated chloride concentrations reduced the number of species of

phytoplankton (Dunlop et al. 2005), along with diminishing the riparian vegetation buffer

system of a stream (Coring and Bäthe, 2011). Increases in salinity have also been

associated with biodiversity shifts in microbial biofilms (Nuy et al., 2018). Biofilms are

complex systems that can make a protected environment for microbes in harsh

environments such as high salinity (Hall-Stoodley et al., 2004).

The U.S. Environmental Protection Agency has set ambient water quality criteria

for chloride associated with sodium, as road salt is usually composed of sodium chloride

(Tiwari et al., 2018). These acute and chronic criteria for chloride are 860 mg Cl-/L and

230 mg Cl-/L, respectively. If these criteria are exceeded, there can be negative impacts

on organisms. Chloride concentrations in many urban streams and rivers in the United

States exceed these criteria (Corsi et al., 2010). Chloride concentrations in urban streams

usually exceed those of streams in forest watersheds, which typically have concentrations

below 50 mg Cl-/L (Corsi et al., 2015, Findlay et al., 2011).

Decomposition is a fundamental process that facilitates nutrient cycling and

supports food webs in freshwater ecosystems that may be affected by increases in chloride concentration. However, relatively little is known about the impact of elevated

chloride concentrations on decomposition and decomposers. In a previous study that

focused on multicellular decomposers and microbes there, was reduced biodiversity due

to increased chloride concentration (Canhoto et al., 2017). Some studies have shown that

decreased biodiversity has little effect on the decomposition rate (Geraldes et al., 2012),

while other authors have reported decreased biodiversity was associated with decreased

decomposition rate (Gonçalves et al., 2015). Elevated chloride concentrations can impact

the structure and function of fungal and bacteria communities. (Canhoto et al., 2017, Nuy et al., 2018). Molecular approaches including high-throughput sequencing techniques have made it possible to describe microbial communities. Canhoto et al. (2017) showed in a lab experiment that chloride concentrations up to 16 mg L-1 caused a decrease in fungal diversity. Nuy et al. (2018) performed a mesocosm experiment focused on how stressors like salinity can affect microbes in streams. Chloride concentrations were experimentally increased up to 312.2 mg L-1 in a mesocosm experiment. They used amplicon sequencing and determined the increase in chloride concentration increased the taxa richness of the microbes.

I conducted the study described here to illustrate how elevated chloride concentrations impact the performance and diversity of decomposers in freshwater systems. Three laboratory experiments were performed where chloride concentration in microcosms were manipulated. Experiments I and II focused on microbes, while experiment III focused on Gammarus pseudolimnaues amphipods, a common shredder in streams. My predictions for experiment I were that: 1) microbial respiration rate will decrease with increasing chloride concentration, 2) percent leaf disk mass remaining (a measure of decomposition) will increase with increasing chloride concentration, and 3) dissolved organic carbon will not be affected. My predictions for experiment II were that:

1) increased chloride concentration will impact bacteria community composition, favoring taxa that are more halophilic. My predictions for experiment III were that: 1) amphipod mortality will increase with increasing chloride concentration, 2) amphipod

instantaneous growth rate will decrease with increasing chloride concentration, and 3)

amphipod egestion rate will increase with increasing chloride concentration.

Methods

2.0 Experimental preparation: water and leaf collection. I performed three

experiments to test my predictions about how elevated chloride would impact

decomposers. The first two focused on microbes and the last one focused on amphipods.

In all experiments, chloride concentration was manipulated in microcosms placed in an

incubator at 13ºC (Table 1). The water and decomposer organisms used in the

experiments were collected from Emmons Creek, a trout stream in Portage County, WI

and the temperature of the incubator reflected the cold-water status of the stream. The

water was collected once in May, June, and July 2018 at the start of each experiment and

the amphipods were collected in June 2018. Emmons Creek was selected because

ambient chloride concentrations are consistently low (i.e. < 4 mg Cl-/L) (Stelzer, personal

communication). Stream water was filtered through 5-µm mesh bags. Dried red maple

(Acer rubrum) leaves were collected at abscission from the Emmons Creek watershed.

Leaves were placed in 1-mm mesh bags (about 12 g leaf/bag) and deployed in Emmons

Cr. for about 2 weeks for microbial colonization. Leaves were removed from the stream and brought back to the lab in a cooler immediately prior to each experiment. After macroinvertebrates were removed, leaves were cut into disks (about 1.2 mm diameter) using a sterilized hole punch. Some disks were placed at -20 °C to estimate the average initial leaf mass.

Leaf disks were placed in microcosms that contained the following chloride treatment solutions ~3 mg Cl-/L (ambient), 50 mg Cl-/L, 300 mg Cl-/L, 1000 mg Cl-/L, and 2500 mg Cl-/L (Table 1). The treatment solutions were prepared by adding reagent

grade NaCl (Ward’s Science, Rochester, New York) to filtered stream water except in the

case of the ambient treatment which did not receive additional salt. The specific

conductance of the solutions was measured with a YSI 30 conductivity meter (YSI

Incorporated, Yellow Springs, Ohio, US) to verify that the appropriate amount of salt had

been added (Table 1). The Cl- concentrations were chosen based on environmental

concentrations reported in stream ecosystems that other authors observed which ranged

from under 100 mg Cl-/L to above 3000 mg Cl-/L (Corsi et al., 2010). Details for each

experiment are provided below.

Table 1

Specific Conductance data from the chloride treatment solutions. There was no 1000 mg Cl-/L treatment on 9 August 2018.

Dates Specific Conductance was measured Treatments 6/5/18 6/20/18 6/29/18 7/6/18 8/9/18 (mg Cl-/L) (µS/cm) (µS/cm) (µS/cm) (µS/cm) (µS/cm) 3 424 409 382 412 438 50 510 515 497 550 542 300 1190 1386 1270 1202 1499 1000 3289 3585 3255 3098 -- 2500 7400 7950 7250 7200 7630

2.1 Experiment I: microbial respiration and decomposition. This experiment was set up by adding 20 leaf disks to autoclaved Erlenmeyer flasks that contained 200 mL of the appropriate chloride solution. The flasks were placed in the incubator in the dark for 2 weeks. There were 10 replicates per treatment. The flasks were aerated gently with aquarium pumps to prevent anoxia. Respiration rates were measured on day 6 of the experiment as follows. Four to six leaf disks were removed from 6 replicates per treatment and placed in 60 mL BOD bottles filled with water from the treatment solutions. Dissolved oxygen concentration in the bottles was measured at 2 hours intervals for 4 hours using a YSI 52 dissolved oxygen meter (YSI Incorporated, Yellow

Springs, Ohio, US) equipped with a stirring probe. The bottles were placed in the incubator (13 °C) in the dark between measurements. At the end of the respiration incubations the leaf disks were stored at -20 °C. Their mass was measured after drying at

60 °C for 24 hours. Respiration rate was determined by linearly regressing oxygen concentration on time and was normalized to leaf dry mass.

On day 14 of the experiment, any remaining leaf disks were removed from the microcosms and stored at -20 °C. These disks were later dried and weighed with a

Mettler Toledo ultra-microbalance (1-µg resolution) (Mettler Toledo, Columbus, Ohio,

US). Their masses were compared to the mean mass of initial leaf disks to estimate the extent of decomposition that had occurred.

After all leaf disks were removed, 100 mL of water from two microcosms per treatment was filtered through Whatman GF/F filters (GE Healthcare Bio-Sciences,

Pittsburgh, Pennsylvania, US) and stored at -20 °C. Dissolved organic carbon (DOC)

concentration of these samples was measured with a Shimadzu SSM-5000A TOC-L

Analyzer (Shimadzu Corporation, Kyoto, Japan) using a high-temperature combustion

method (American Society for Testing and Materials, 1994).

2.2 Experiment II: bacterial diversity. The setup of this experiment was

identical to that of experiment I except the focus was on bacterial diversity instead of

microbial respiration. Sixty leaf disks were placed in Erlenmeyer flasks containing 200

ml of filtered stream water. Treatments, each replicated 5 times, consisted of 3 mg Cl-/L,

50 mg Cl-/L, 300 mg Cl-/L, and 2500 mg Cl-/L. After 10 days, the leaf disks were

collected using sterile technique and stored at -20 °C. DNA was extracted from 2

replicates per treatment based on the methods of Das et al., (2007). About 150 mg (wet

mass) of leaf material was freeze dried with liquid nitrogen and then homogenized with a mortar and pestle. A PowerSoil DNA Extraction Kit (Qiagen Sciences Inc, Germantown,

Maryland, US) and a DNA Clean and Concentrator Kit (Zymo Research, Irvin,

California, US) were used for DNA extraction and purification following manufacturer instructions. The quantity and quality of the DNA were assessed using a Nanodrop

Spectrophotometer ND-1000 (Thermofisher Scientific, Waltham, Massachusetts, US) and

Invitrogen Qubit 2.0 Fluorometer (Thermofisher Scientific, Waltham, Massachusetts,

US). PCR was performed with electrophoresis gel using the 8 forward and 1492 primers to confirm there was no inhibition (Frank et al., 2008).

The community composition of the bacteria associated with the leaf disks (2 replicates per treatment) was measured with Illumina sequencing of the 16S rRNA targeting the V3-V6 region. The amplicon sequencing and basic assembly and analysis of

the data were performed by Molecular Research (Mr. DNA, Shallowater, Texas, US).

Bacteria identity was determined at Molecular Research by comparing sequence data to a

database from the NCBI (Dowd et al., 2008). Relative abundances, Shannon-Weiner

Diversity Index, and Genera Evenness were calculated using these genus-level data.

2.3 Experiment III: amphipod performance. In this experiment, amphipods

were exposed to the same chloride treatments as described in experiment I. Amphipods

(Gammarus sp.) were collected from Emmons Creek and brought to the lab where their

initial blotted wet mass was measured with a Mettler Toledo ultra-microbalance (1 µg

resolution). Only amphipods within the range of 2 to 7 mg were used. Amphipods were

placed separately in plastic containers that contained 100 mL of filtered stream water and two leaf disks. A completely randomized design was employed with 18 replicates per treatment. Amphipods were placed in the incubator which was set to a 12-hour light cycle

(12 hours light:12 hours dark). At two-day intervals the amphipods were given fresh

chloride treatment water, the old leaf disks were removed and stored at -20 °C and new

disks were added. Amphipod mortalities were noted during the water changes.

On day 14 of the experiment, the blotted wet mass of the amphipods was measured and instantaneous growth rate (G) was calculated with the equation:

G = ln (( / )/ ) Eq. 1. 𝑡𝑡 0 where: 𝑀𝑀 𝑀𝑀 𝑡𝑡

is the amphipod mass at time (t),

𝑡𝑡 𝑀𝑀 is the initial amphipod mass,

0 and𝑀𝑀 t is the number of days elapsed between the initial and final measurements.

After the amphipods were weighed on Day 14, they were returned to their

containers, which contained new leaf disks, and allowed to feed for 72 hours. On Day 17, after leaf disks and amphipods were removed, the remaining water from the microcosms was filtered through pre-weighed Whatman GF/F filters in order to collect the fecal pellets. The filters were stored at -20 °C, subsequently dried at 60 °C, and reweighed.

Egestion rates (E) were calculated from these data based on the following equation:

= ( )/ / Eq. 2.

𝑡𝑡 0 where: 𝐸𝐸 𝑊𝑊 − 𝑊𝑊 𝐴𝐴 𝑡𝑡

is the initial filter weight,

0 𝑊𝑊 is the filter weight after time (t),

𝑡𝑡 A𝑊𝑊 is the amphipod blotted wet mass,

t is the incubation duration in days.

2.4 Data analysis. Analysis of variance (ANOVA) was used to compare means of

the response variables, including respiration rate and instantaneous growth rate, among

the treatments. Tukey tests were used to compare means of individual treatments if the

overall ANOVA models were statistically significant. A probit analysis was used to find

the LD for the amphipods (Randhawa 2009). A Shannon-Weiner Diversity Index was

50 used to assess microbial diversity. All statistical analysis was performed in R.

Results

3.1 Experiment I: microbial respiration and decomposition. There was no significant difference in mean microbial respiration rates among the chloride treatments

(One-way ANOVA, p=0.6308, Figure 1). The respiration rates decreased slightly as the chloride concentrations increased, but this result was not significant.

There was also no significant difference in the mean amount of leaf mass remaining among treatments for each day that it was measured (One-way ANOVA, day

6: p=0.3688, day 14: p=0.9585, Figure 2).

0.12

0.10

0.08 /mg leaf/min) /mg 2

0.06

0.04

0.02 Respiration Rate (mg O (mg Rate Respiration

0.00 1 10 100 1000 (mg Cl-/L) Figure 1. Microbial respiration rates (means+SD) after exposure to the chloride treatments.

140 3 50 120 300 1000 2500 100

40 PercentDisk Leaf MassRemaining 20

0 6 14 Days Figure 2. Percent leaf disk mass remaining on Day 6 (6/6/18, Day 1) and Day 14 (6/14/18, Day 2) (means+SD).

16 b ab 14

12 ab ab 10

8 a

DOCC/L) (mg 6

0 1 10 100 1000 - (mg Cl /L) Figure 3. Dissolved organic carbon (DOC) (means+SD). (a) and (b) represent significance. Bars with different letters indicate that means are different in pairwise comparisons.

Mean DOC concentrations differed among chloride treatments (One-way

ANOVA, p=0.035, Figure 3). The 1000 mg Cl-/L treatment had a higher mean

concentration of DOC than the 3 mg CL-/L treatment (Tukey HSD, p=0.0382). The 3

mg Cl-/L treatment had the lowest concentration of DOC compared to the other

treatments.

3.2 Experiment II: bacterial diversity. Bacterial community composition, based

on genera relative abundances, differed among chloride treatments, especially when the

lowest concentration treatments (3 and 50 mg Cl-/L) were compared to the highest

concentrations (300 and 2500 mg Cl-/L) (Figure 4). Most bacteria were Bacteroidetes and

Proteobacteria (Appendix IV). Several bacterial genera (e.g. Flavobacterium,

Rhizobium, Acidovorax) were more prominent at lower chloride concentrations while others (e.g. Aeromonas and Lactobacillus) were more prominent at higher chloride concentrations (Figure 4 and 5). Five hundred and three total bacteria genera were identified. The genera cut off was 3% divergence (97% similarity) and the number of

sequence counts was 86000-126000. One hundred-forty bacteria genera were only found in the lowest chloride treatment. Nineteen genera were only found in the highest chloride treatment (Appendix I). One hundred and fifty-five genera were found in all treatments

(Appendix V, Supplemental Figure 1). Shannon Diversity Index (H’) differed among treatments (Table 2). The highest H’ average (3.771) was in the 50 mg Cl-/L treatment

and the highest genera evenness average (0.698) was also in the 50 mg Cl-/L treatment.

The highest genera richness occurred in the 3 mg Cl-/L treatment with an average of 325

different genera and the lowest average genera richness occurred in the 2500 mg Cl-/L

treatment with 201. The total counts of amplified sequences were similar among

treatments (Figure 5).

Variation between replicates in the relative abundances of bacteria differed among

treatments. Both the 3 and 50 mg Cl-/L treatments had little variation between replicates

while there was high variation between replicates in the 300 and 2500 mg Cl-/L

treatments. The 3 mg Cl-/L treatment had 13 genera that exceeded the 1.5% relative

abundance threshold in one replicate and 10 genera in the second. In this treatment there

were similar relative abundances between replicates for most genera. For example,

Flavobacterium was at 31.6% and 32% in the first and second replicate in the 3 mg Cl-/L

treatments and decreased in relative abundance to 26.2% and 11.0% in the 2500 mg Cl-/L

treatment. Rhizobium also decreased from similar relative abundances of 11.0 and 9.2%

in the 3 mg Cl-/L treatment to 3.1% and below 1.5% in the 2500 mg Cl-/L treatment

(Figure 4). The 50 mg Cl-/L treatment also had many taxa that had similar relative

abundances between the two replicates. Flavobacterium was at 20.1 and 19.7% in the

first and second replicate and Rhizobium was at 6.2% relative abundance for both

replicates. (Figure 4). In the 300 mg Cl-/L treatment, Aeromonas was present in the first

replicate at 50.5% while it was below 1.5% in the second replicate. In the replicate with

more Aeromonas, there were also only five genera present that exceeded the 1.5%

threshold while the other replicate had eight genera present above 1.5%. The 2500 mg Cl-

/L treatment also had high variation between replicates. There were 11 different genera in

the first replicate compared to four in the second replicate. The second replicate had a

much higher amount of Aeromonas (62.7%), but a lower amount of Flavobacterium

(11.0%), Lactobacillus (10.1%), and Arthrobacter (2.1%) compared to the other replicate.

Table 2

Shannon-Weiner Diversity Index, Genera Evenness andRichness (mean ± SD) with all taxa in chloride treatments included

Treatments (mg Cl- Shannon-Weiner Genera Evenness Genera /L) Index (H) Richness 3 3.251 ± 0.06 0.562 ± 0.01 325.5 ± 12.02 50 3.771 ± 0.04 0.698 ± 0.004 221.5 ± 4.95 300 2.94 ± 1.01 0.537 ± 0.182 237 ± 9.90 2500 2.747 ± 0.10 0.425 ± 0.16 201.5 ± 12.02

20 Aeromonas Aeromonas Cellulomonas Cellulomonas Uliginosibacterium Asticcacaulis Pseudomonas Methylibium Brevundimonas Halospirulina Buttiauxella Rhodobacter Rhodoferax Massilia Luteolibacter Ferruginibacter Delftia Arthrobacter Methylotenera Cloacibacterium Novosphingobium Lactobacillus Klebsiella Rhizobium Rhizobium Acidovorax Flavobacterium Flavobacterium 2500 300 50 Treatments (mg Cl-/L) (mg Treatments 3

Figure 4. Bacteria genera relative abundance after exposure to chloride treatments. Bacteria Bacteria treatments. chloride to exposure after abundance relative genera 4. Bacteria Figure per bars The separate rRNA. 16S of sequencing amplicon on based composition community abundance relative 1.5% exceeded that genera those Only replicates. indicate treatment are shown. (per replicate) 0

80 60 40 20 100 (%) Abundance Relative

21 Klebsiella Klebsiella Aeromonas Rhodobacter Rhodobacter Rhodoferax Massilia Luteolibacter Ferruginibacter Delftia Arthrobacter Methylotenera Cloacibacterium Novosphingobium Lactobacillus Halospirulina Halospirulina Buttiauxella Methylibium Methylibium Brevundimonas Cellulomonas Cellulomonas Uliginosibacterium Asticcacaulis Pseudomonas Flavobacterium Flavobacterium Rhizobium Acidovorax 2500 300 50 Treatments (mg Cl-/L) (mg Treatments 3

Figure 5. Bacteria genera sequence counts after exposure to chloride treatments. Bacteria community community Bacteria treatments. chloride to exposure after counts sequence genera 5. Bacteria Figure per treatment bars The separate rRNA. 16S of sequencing amplicon on based composition (per replicate) abundance relative 1..5% exceed that genera those Only replicates. indicate are shown. 1.2e+5 1.0e+5 8.0e+4 6.0e+4 4.0e+4 2.0e+4 0.0 Genera Sequence Counts Sequence Genera

100

40 PercentMortality (%)

0 1 10 100 1000 (mg Cl-/L) Figure 6. Amphipod percent mortality after exposure to chloride treatments.

0.025 ab a

0.020 ab

0.015 b

0.010

0.005 ab

0.000 Instantaneous Growth Rate (1/day) Instantaneous -0.005

-0.010 1 10 100 1000 (mg Cl-/L) Figure 7. Amphipod instantaneous growth rate (means+SD).

3.3 Experiment III: amphipod performance. Amphipod performance generally

decreased as chloride concentrations increased. Percent mortality increased at higher

chloride concentrations (Figure 6). There was 100% mortality in the 2500 mg Cl-/L treatment. All the amphipods in this treatment perished within two weeks. Mortalities occurred throughout the experiment (Appendix V, Supplemental Figure 2). For example, out of 18 initial amphipods in the 2500 mg Cl-/L treatment, seven died during the first

week of the experiment and eleven died during the second week (Appendix V,

Supplemental Figure 2). The mortalities did not differ among the 3, 50, and 300 mg Cl-/L treatments. The LD50, determined with the probit analysis at 14 days, was 1015 ±186 mg

Cl-/L.

Instantaneous growth rate of Gammarus was lower at the highest chloride

concentrations. (One-way ANOVA, p=0.016, Figure 7). Mean growth rate differed

0.25

0.20

0.15

0.10

mg dry mass/mg amphipod/day dry mg 0.05

0.00 1 10 100 1000 - (mg Cl /L) Figure 8. Amphipod egestion rates (means+SD).

between the 300 mg Cl-/L and the 1000 mg Cl-/L treatments (Tukey HSD, p=0.0209).

Because of the high mortality in the 2500 mg Cl-/L treatment, there were only two amphipods for which Fgrowth rate could be reported (these 2 amphipods died shortly after their final masses were measured for the growth rate determinations). This resulted in reduced statistical power to detect growth differences between the highest chloride concentration treatment and the other lower chloride treatments.

Egestion rate was not significantly affected by chloride concentration (One-way

ANOVA, p=0.676, Figure 8). Egestion rate was not calculated for any amphipods in the

2500 mg Cl-/L treatment because all amphipods had died in this treatment before egestion rate was measured at 14 days.

Discussion

Most predictions for all three experiments were supported. As expected, there was a shift in the bacterial community composition with increasing chloride concentration.

Amphipod mortality increased with elevated chloride concentration while amphipod instantaneous growth rate decreased. Contrary to predictions, microbial respiration rate, percent leaf mass remaining, and amphipod egestion rate were not affected by chloride concentration. DOC also increased with increasing chloride concentrations.

Microbial responses. Microbial respiration rate was not affected by the chloride treatments. This may have been because of a bacteria community composition shift.

Although bacterial community composition was not measured in experiment I, the similar experimental conditions between experiment I and II suggest that the bacterial

communities in experiment I also shifted in response to elevated chloride concentration.

The respiration rate may not have been affected because of functional redundancy in the

microbial communities, even though there were likely shifts and differences in

community composition. My results differed from other studies that have examined the impact of elevated chloride concentration on microbial respiration (Tyree 2016, Canhoto et al. 2017). For example, Tyree (2016) found that chloride concentrations exceeding 200 mg Cl-/L caused a decrease in microbial respiration rates after 28 days compared to

bacteria exposed to lower chloride concentrations. Tyree (2016) suggested that the

respiration rate could have decreased at higher chloride concentrations due to

osmoregulatory stress. The percent leaf disk mass remaining, which was an indirect

measure of decomposition, was also not significantly affected by the higher chloride

treatments. A change in biomass may have possiblyF masked any changes in

decomposition that occurred. Other microcosm studies have found a lower decomposition

rates at higher chloride concentrations. Canhoto et al (2017) found that leaf

decomposition of leaves was negatively affected by increases in salinity up to 1600 mg /L

NaCl after 35 days which does not agree with my results. In contrast, Tyree (2016) found

that decomposition was not impacted by Cl concentrations up to 214.7 mg/L which

agreed with my results. The length of my experiment was shorter compared to both these

studies since it was only 14 days which may have been a factor on why there was no

significant change in respiration or decomposition.

There were major shifts in microbial community composition as chloride

concentration increased. Several taxa became less prominent as chloride concentration

increased. The decreased prominence of genera like Flavobacterium and Rhizobium at

higher chloride concentrations may have been due to a lack of salt tolerance by these

taxa. Genera that did increase in relative abundance in the higher chloride treatments

included Aeromonas and Lactobacillus which increased from <1.5% in the 3 and the 50

mg Cl-/L treatment to 62.7% and 23.6% in the 2500 mg Cl-/L treatment respectively

(Figure 4). This increase in gene sequences of genera such as Aeromonas and

Lactobacillus may have been because these genera were halophilic or halotolerant.

Bacterial species that are halophilic or halotolerant can maintain an osmotic balance by regulating their organic solutes and accumulating high concentrations of various solutes when necessary (Margesin 2001). Aeromonas and Lactobacillus have been shown to be

halophilic and halotolerant in previous studies (Batra et al., 2016, Hutkins et al., 1987).

Another genus that increased in relative abundance at the higher chloride treatments was

Arthrobacter which increased from <1.5% relative abundance in the 3 mg Cl-/L treatment

to 4.4% in the 2500 mg Cl-/L treatment. Arthrobacter has been found in moderately saline soils in previous studies (Mukhtar et al., 2018). Microbial community shifts with an increase in chloride concentration has been seen in other studies. Canhoto et al. (2017) found shifts in the fungal community from higher to lower diversity with increasing chloride concentrations. Nuy et al. (2018) also showed that an increase in salt influenced the number of OTUs and taxa richness of bacteria and fungi, but the richness increased with the increase in salinity. This may have been because the increase in chloride concentrations was relatively low (18.2 mg Cl-/L to 312.2 mg Cl-/L).

Many of the bacteria genera identified in experiment II were restricted to one chloride treatment. The large number of bacterial genera only found in the 3 mg Cl-/L

treatment and decrease in unique genera at higher chloride concentrations may have been

due to the relatively low osmotolerance of these genera. The genera at higher chloride

concentrations may have had higher osmotolerance. For example, Synechococcus was

one of the genera restricted to the 2500 mg Cl-/L treatment. This genus has been found in hypersaline microbial mats. These mats can be found in deserts and taxa such as

Synechococcus that are tolerant to high salinities and desiccation can survive in them by changing the solutes and fatty acids they produce (Abed et al., 2015). These hypersaline microbial mats can sometimes include bacteria such as Chroococcidiopsis (Abed et al.,

2015), an extremophile, which were only found in the 300 mg Cl-/L treatment in this

experiment.

The shift in bacterial community composition with increasing chloride

concentration did not appear to impact overall metabolic function and decomposition of

the bacterial community since there was no effect of chloride concentration on respiration

rate or decomposition in exp I. Decomposition and other functions such as respiration

rate may not be affected since there can be functional redundancy in communities, which

means that measures of community function such as respiration are not necessarily

affected if communities change (Shade et al., 2012). There were also genera that were

present in all treatments that could have been resilient to the chloride increase. This

resiliency and the interactions and responses between this part of the bacterial community

could have been involved with the functional redundancy to help the decomposition and

respiration rates remain the same. My results are consistent with those of Canhoto (2017)

who found that a loss of fungal species at a higher salt concentration did not depress leaf decomposition (Canhoto et al., 2017). I realize that there are many different biogeochemical processes (e.g. nitrogen transformation, enzyme-driven carbon transformation) that may have changed as the bacteria communities shifted at high salt concentration, but were not evaluated in this study.

The DOC concentration may have been higher in the treatments with higher chloride concentration because of the change in microbial community composition and this change could affect carbon processing or because of the elevated chloride in the water directly causing the DOC to leach out of the leaf disks. A few studies have observed that increased salt exposure can lead to leaching organic matter from soils

(Green et al., 2009, Steele et al., 2013). Steele et al (2013), found that sodium had a large impact on DOC concentration in solutions with increasing sodium adsorption ratios, leading to higher leaching. The amount of leaching that occurred depended on the type of vegetation along with the amount of salinity and sodicity (relative proportion of sodium compared to other cations) (Steele et al., 2013)

Gammarus performance. Elevate chloride concentrations had lethal and sublethal effects on amphipods. The higher mortality in the treatments with elevated chloride concentration may have been a result of osmoregulatory stress. Chloride can be lethal if organisms are not able to handle the change in osmotic pressure (Kefford et al.,

2016). The increase in mortality was expected because other investigators have determined that LD s of amphipods ranged from 1,276 mg Cl-/L to 1,496 mg Cl-/L

(Elphick et al., 2011), which is similar to the LD of 1015. ±186. mg Cl-/L found in this

50 experiment. The 2500 mg Cl-/L treatment was more than double these LD s and was

50 almost three orders of magnitude higher than what the amphipods are exposed to in

Emmons Creek. Hence, the highest mortality in this treatment was expected.

Instantaneous growth rate declined at the highest chloride concentrations, which suggests sub-lethal osmotic or chloride stress. Many species of amphipods have specialized cells near there gills that are known to be mitochondria-rich (Matsumasa et al., 1998). These specialized ion-transport cells are similar to chloride cells in teleost gills and salt-excreting cells in the gills of brine shrimp. These cells facilitate osmoregulation in amphipods and it has been suggested they are involved in the excretion of sodium chloride in hyperosmotic environments (Matsumasa et al., 1998). Aquatic invertebrates can regulate their haemolymph by producing more amino acids and proteins when they need to prevent dehydration and cell shrinkage. In addition, concentrating their haemolymph relative to a hypotonic environment can help invertebrates retain water to prevent dehydration and possible cell shrinkage (Tiwari et al., 2018). Other investigators such as Tyree (2016) have found that somatic growth in Lirceus sp. decreased when energy is allocated to making sure organisms are able to regulate the osmotic balance instead of growth. This regulation in hypertonic conditions tends to require more energy since ions are transported by active transport across membranes than some organisms can acquire by consumption. Increased energy demands by amphipods in the high chloride concentration treatments might have led to decreased growth and increased death in the highest chloride treatments.

Egestion rate of amphipods was not significantly affected by elevated chloride

concentration. The egestion rate may have been unaffected while the instantaneous

growth rate decreased in higher chloride concentrations. If the amphipods were ingesting

leaf material at a similar rate among treatments but were not able to allocate energy to new tissue production in the higher salt concentrations. Egestion not being affected with increases in chloride concentration was also found in a study done by Tyree (2016).

Tyree’s study measured assimilation efficiency which takes egestion into account along with consumption. Assimilation efficiency of Lirceus isopods was estimated in microcosms, and Tyree found that there were no significant changes in assimilation efficiency with increases in chloride concentration (Tyree et al., 2016).

4.4 Summary. Elevated chloride concentrations lead to major changes in bacterial community composition and amphipod performance. DOC concentration increased at higher chloride concentrations and bacterial community composition shifted to more halophilic genera. Microbial respiration rate and decomposition were not affected by chloride concentration. Amphipod mortality increased with increasing chloride concentration while amphipod instantaneous growth rate decreased. Amphipod egestion was not affected. Collectively, these results suggest that as chloride concentration in surface water increases (Gardner et al., 2010, Wallace et al., 2016, Chapra et al., 2012), the community composition of benthic bacteria and the amphipod performance will be impacted, which may have implications for decomposition rates in aquatic ecosystems.

These results are relatively novel because studies of how chloride concentration impacts microbial communities involved in decomposition in aquatic systems are relatively rare.

A fruitful direction for future research would consider how shifts in bacterial community

composition in response to anthropogenic changes affect biogeochemical processes at the decomposer–organic carbon interface.

Chapter III

Conclusions

The microbial results of my study were that the community composition of

bacteria experienced a shift while the microbial respiration and percent leaf mass

remaining (a measure of decomposition) rates were not affected and the dissolved organic

carbon (DOC) increased. The amphipod results showed that amphipod mortality

increased to 100% in 2 weeks in the 2500 mg Cl-/L treatment while the amphipod

instantaneous growth rate decreased, and the amphipod egestion rate was not affected.

The microbial results showed that microbial function did not seem to be affected

by the high chloride concentrations, while the community composition was affected.

Microbial respiration not being significantly affected was unexpected based on the results

of previous studies (Tyree et al., 2016). There could have been no effect because of the

functional redundancy or multiple organisms able to do the same function in the

microbial community. This allows functions such as respiration in this case to not

necessarily be affected if there is a change in the community (Shade et al., 2012). The shift in the bacteria community was predicted. Some taxa in the higher chloride concentration treatments were salt tolerant (Mukhtar et al., 2018, Batra et al., 2016,

Hutkins et al., 1987) and were able to survive in these higher chloride concentrations better than other could. Some of the taxa that were in the higher chloride treatments may have been the cause of the increase in DOC if they processed carbon differently compared to the bacteria in the lower chloride concentration treatments. The increase in

DOC may also have been directly caused by the higher chloride concentrations allowing the DOC to leach out of the leaf disks more.

The amphipod results suggested that chloride can affect the performance of amphipods. There was an increase in mortality in the higher chloride concentration treatments, which was expected since the chloride concentration levels were picked to be higher than the amphipods were used to so they would cause osmoregulatory stress that could be lethal the amphipods (Corsi et al., 2015). This stress could also be the reason for the lower instantaneous growth rates. Amphipods have specialized cells near their gills that help them excrete salt and osmoregulate and can also regulate their haemolymph to prevent dehydration (Matsumasa et al., 1998, Tiwari et al., 2018). The amphipods may have been allocating most of their energy to regulating their osmotic balance through these methods instead of their growth which may be why it decreased in the higher chloride concentration treatments. The egestion rate was not significantly affected by the increase in chloride. This may have been because the amphipods in all treatments were ingesting leaf material at a similar rate, but those in the higher chloride treatments may have been less efficient at allocating energy to new tissue production, so the egestion rates did not differ between treatments.

These results help show how chloride can impact microbial communities and amphipods that are involved in decomposition. Learning how these organisms are affected can improve knowledge about salt toxicity which is important since there have been increases in chloride concentration in surface waters around the world (Gardner et al., 2010, Wallace et al., 2016, Corsi et al., 2015, Chapra et al., 2012). This increase is not

expected to stop since road salt use will continue and will have impacts on aquatic ecosystems near urban areas and on humans since water with higher chloride concentration levels can be toxic to organisms and are not considered potable for human consumption (Cañedo-Argüelles et al., 2013, Kaushal et al., 2018). Using the current knowledge about road salts and chloride more communities are becoming increasingly proactive about what is in their water and will continue to need new information to help make informed decisions about how to keep their communities and freshwater ecosystems functioning well.

APPENDIX A

Genera of bacteria that were present exclusively in individual chloride treatments

Treatments (mg Cl-/L) 3 50 300 2500 Acanthamoeba Chitinimonas Archaeoglobus Anaerococcus Acidiferrobacter

Acetivibrio Chloroidium Caldilinea Candidatus chloracidobacterium Alteromonas

Acetobacterium Chondromyces Candidatus acetothermum Candidatus nitrososphaera Atopostipes

Acidimicrobium Citreicella Ehrlichia Carboxydothermus Brooklawnia

Acidisphaera Cohnella Estrella Chroococcidiopsis Candidatus kuenenia

Adhaeribacter Cupriavidus Ignavibacterium Collinsella Candidatus planktophila

Alcanivorax Cyanidium Isosphaera Frateuria Capnocytophaga

Alkanindiges Cystobacter Methanosphaera Frondihabitans Globicatella

Alloactinosynnema Dehalococcoides Methylomicrobium Halochromatium Heliospora

Allokutzneria Dehalogenimonas Mucilaginibacter Isoptericola Marinithermus

Aminobacter Desulfobacca Pasteuria Loktanella Meiothermus

Ancalomicrobium Desulfotalea Roseibacterium Microbulbifer Nitrososphaera

Azoarcus Diaphorobacter Zea Nitrosospira Olsenella

Azospira Diplorickettsia Nitrospirillum Parvibaculum

Azospirillum Dongia Odoribacter Planococcus

Bdellovibrio Dyella Peptoniphilus Schlegelella

Beggiatoa Ectothiorhodospira Peptostreptococcus Solirubrobacter

Brenneria Elstera Pleomorphobacterium Synechococcus

Buttiauxella Epulopiscium Polyangium Tepidimicrobium

Candidatus babela Ferrimicrobium Rubellimicrobium

Candidatus captivus Fibrella Singulisphaera

Candidatus carsonella Fibrobacter Thiomicrospira

Candidatus lariskella Flectobacillus Williamsia

Candidatus nucleicultrix Fluviimonas

Candidatus trichorickettsia Formivibrio

Candidatus xiphinematobacter Fusobacterium

Catenuloplanes Gallionella

Chamaesiphon Geobacillus

Gordonibacter Oxalobacter

Haematobacter Paludibacter

Haliangium Paludibacterium

Helicobacter Pantoea

Herminiimonas Paracraurococcus

Humitalea Parvularcula

Kaistia Pedomicrobium

Kitasatospora Pelobacter

Labrys Phormidium

Treatments (mg Cl-/L)

3 50 300 2500 Lachnoclostridium Phyllobacterium

Lacibacterium Picochlorum

Leptolyngbya Planomicrobium

Levilinea Pleomorphomonas

Luedemannella Propionigenium

Magnetospirillum Prototheca

Marinobacter Pseudolabrys

Metallibacterium Quatrionicoccus

Methylobacillus Rahnella

Methylomonas Rheinheimera

Methylovorus Rhizobacter

Micavibrio Rhizomicrobium

Modestobacter Rhodocyclus

Morus Rhodopseudomona s Myxozyma Rhodovastum

Nakamurella Roseimaritima

Nitratireductor Roseobacter

Nocardiopsis Saccharibacillus

Novispirillum Saccharospirillum

Ochrobactrum Sanguibacter

Opitutus Sideroxydans

Simaetha Thauera

Sinorhizobium Thermanaerothrix

Smithella Thermovum

Solibacillus Thiorhodospira

Sporobacter Vibrio

Sulfurospirillum Xylella

Tabrizicola Yonghaparkia

Terrabacter Zebrasoma

Thalassospira Zoogloea

APPENDIX B

Bacteria sequence count data for all treatments (Two replicates are indicated per treatment)

Treatments (mg Cl-/L) Genera 3 50 300 2500 Acanthamoeba 0 16 0 0 0 0 0 0

Acetivibrio 0 6 0 0 0 0 0 0

Acetobacterium 17 75 0 0 0 0 0 0

Acidimicrobium 10 3 0 0 0 0 0 0

Acidisphaera 20 7 0 0 0 0 0 0

Acidiferrobacter 0 0 0 0 0 0 11 0

Aciditerrimonas 26 43 0 0 141 1 71 51

Acidobacterium 37 22 398 84 159 166 0 2

Acidothermus 0 0 1 0 0 2 0 0

Acidovorax 4463 6483 4081 2465 1139 682 1999 1209

Acinetobacter 34 18 290 327 463 59 848 30

Actinomadura 0 4 1 0 347 1 0 1

Actinomyces 0 0 2 29 64 3 0 0

Actinomycetospora 4 5 0 0 20 68 0 0

Actinoplanes 183 468 1183 398 175 40 764 13

Adhaeribacter 4 0 0 0 0 0 0 0

Aerococcus 0 0 1 0 0 108 0 0

Aeromonas 953 425 572 284 420 63667 549 63711

Agrobacterium 741 710 691 687 530 293 254 59

Agrococcus 80 4 95 1 1 0 0 0

Alcanivorax 5 0 0 0 0 0 0 0

Alkanindiges 4 6 0 0 0 0 0 0

Alloactinosynnema 5 0 0 0 0 0 0 0

Allokutzneria 0 4 0 0 0 0 0 0

Algorimarina 0 0 0 167 0 0 0 1

Altererythrobacter 91 108 120 715 3 103 103 66

Alteromonas 0 0 0 0 0 0 0 1

Amaricoccus 18 61 2 1 85 0 119 29

Aminobacter 11 24 0 0 0 0 0 0

Amnibacterium 0 0 161 76 878 481 8 25

Anaerococcus 0 0 0 0 119 68 0 0

Anaerolinea 19 47 41 62 0 49 0 1

Anaeromyxobacter 16 39 1 0 1 2 13 0

Ancalomicrobium 0 3 0 0 0 0 0 0

Aquabacterium 355 260 454 721 67 133 283 76

Aquicella 55 13 89 1 53 1 2 0

Aquimonas 4 8 0 1 1 25 49 1

Treatments (mg Cl-/L) Genera 3 50 300 2500 Aquincola 2 5 0 2 3 0 0 0

Archaeoglobus 0 0 27 0 0 0 0 0

Arcicella 98 221 414 689 101 224 4 4

Arcobacter 7 46 0 172 0 1 0 0

Arenimonas 149 227 399 21 49 42 2 40

Arthrobacter 25 26 1529 1423 5847 3353 3895 2181

Asticcacaulis 2727 3306 1661 862 921 1053 657 333

Atopostipes 0 0 0 0 0 0 0 1

Aurantimonas 0 12 0 0 0 22 0 0

Azoarcus 2 1 0 0 0 0 0 0

Azonexus 11 10 0 121 0 0 1 0

Azorhizobium 9 12 0 0 58 0 0 0

Azospira 9 53 0 0 0 0 0 0

Azospirillum 5 7 0 0 0 0 0 0

Bacillus 66 17 71 156 224 197 109 3

Bacteriovorax 14 47 48 1 116 0 0 1

Bacteroides 14 22 0 1 125 65 0 0

Bdellovibrio 41 45 0 0 0 0 0 0

Beggiatoa 22 29 0 0 0 0 0 0

Beijerinckia 175 149 108 305 48 0 18 2

Bergeyella 0 0 0 0 238 0 0 1

Blastochloris 1 2 0 0 0 32 0 0

Blastococcus 0 0 21 0 247 2 1 0

Blastomonas 0 0 0 1 0 0 77 0

Blastopirellula 16 12 234 30 29 0 1 1

Bosea 88 110 20 55 1 68 0 2

Brachybacterium 7 0 0 1 0 0 1 62

Bradyrhizobium 115 148 1 134 1 2 0 3

Brenneria 0 37 0 0 0 0 0 0

Brevibacillus 4 0 0 0 0 0 0 81

Brevundimonas 2184 2304 741 320 612 716 455 411

Brooklawnia 0 0 0 0 0 0 0 11

Brucella 60 60 63 132 67 36 1 0

Burkholderia 6 18 212 130 175 75 1 0

Buttiauxella 1722 946 0 0 0 0 0 0

Byssovorax 0 3 0 61 90 0 0 0

Bythopirellula 10 0 65 0 1 0 0 1

Treatments (mg Cl-/L) Genera 3 50 300 2500 Caedibacter 41 30 0 92 1 23 0 0

Caldilinea 0 0 0 49 0 0 0 0

Candidatus acetothermum 0 0 0 1 0 0 0 0

Candidatus alysiosphaera 0 0 0 1 74 2 27 0

Candidatus babela 0 4 0 0 0 0 0 0

Candidatus captivus 3 0 0 0 0 0 0 0

Candidatus carsonella 3 1 0 0 0 0 0 0

Candidatus chloracidobacterium 0 0 0 0 39 1 0 0

Candidatus kuenenia 0 0 0 0 0 0 0 4

Candidatus lariskella 3 0 0 0 0 0 0 0

Candidatus metachlamydia 41 55 124 0 0 259 0 1

Candidatus microthrix 18 1 89 84 0 77 3 0

Candidatus midichloria 21 28 65 0 0 0 0 0

Candidatus nitrosoarchaeum 0 0 2 2 3 0 0 0

Candidatus nitrososphaera 0 0 0 0 0 3 0 0

Candidatus nucleicultrix 27 28 0 0 0 0 0 0

Candidatus odyssella 16 31 228 1 1 62 0 2

Candidatus pelagibacter 0 0 17 0 116 3 1 1

Candidatus phytoplasma 61 12 3 254 793 119 400 58

Candidatus planktophila 0 0 0 0 0 0 0 38

Candidatus protochlamydia 10 0 0 0 89 0 0 0

Candidatus rhabdochlamydia 8 2 0 0 0 61 0 0

Candidatus saccharimonas 0 8 22 92 266 41 32 1

Candidatus solibacter 0 0 133 0 2 1 0 0

Candidatus trichorickettsia 3 0 0 0 0 0 0 0

Candidatus vidania 1 2 54 0 1 62 0 0

Candidatus xiphinematobacter 0 3 0 0 0 0 0 0

Capnocytophaga 0 0 0 0 0 0 15 0

Carboxydothermus 0 0 0 0 126 0 0 0

Carnobacterium 4 1 0 0 0 0 43 1

Catellicoccus 0 0 0 0 1 1 126 0

Catenuloplanes 0 9 0 0 0 0 0 0

Caulobacter 548 976 415 281 186 122 111 125

Cellulomonas 4287 1493 478 687 278 414 152 70

Cellvibrio 97 51 136 46 0 55 1 42

Chamaesiphon 7 0 0 0 0 0 0 0

Chelatococcus 26 12 5 108 2 3 40 69

Treatments (mg Cl-/L) Genera 3 50 300 2500 Chitinimonas 7 8 0 0 0 0 0 0

Chitinophaga 203 526 359 6 3 70 0 2

Chloroflexus 3 13 122 2 55 2 79 1

Chloroidium 6 0 0 0 0 0 0 0

Chondromyces 6 1 0 0 0 0 0 0

Chroococcidiopsis 0 0 0 0 219 0 0 0

Chryseobacterium 4 0 38 1 0 0 0 0

Chthoniobacter 4 25 0 93 0 155 1 0

Citreicella 63 32 0 0 0 0 0 0

Cloacibacterium 0 5 1 1623 447 7 1 2

Clostridium 67 74 189 92 219 7 55 6

Cohnella 0 3 0 0 0 0 0 0

Collimonas 0 0 85 0 1 0 0 0

Collinsella 0 0 0 0 30 0 0 0

Comamonas 20 52 53 85 0 1 117 3

Compostimonas 35 13 1 0 1 64 0 80

Conexibacter 22 27 11 0 252 1 46 52

Corynebacterium 1 15 127 922 825 12 7 31

Coxiella 26 21 2 94 0 74 26 2

Crocinitomix 73 62 311 386 79 321 0 4

Cronobacter 0 0 0 0 41 0 1 0

Cryocola 0 0 6 7 461 10 4 8

Cupriavidus 3 2 0 0 0 0 0 0

Curtobacterium 197 34 0 0 18 97 5 0

Curvibacter 4 2 191 79 50 35 49 8

Cyanidium 15 6 0 0 0 0 0 0

Cystobacter 5 0 0 0 0 0 0 0

Cytophaga 127 330 177 303 207 2 56 2

Daeguia 0 0 93 120 70 164 22 17

Dechloromonas 101 122 69 1 0 49 0 2

Deefgea 0 5 75 0 0 0 1 0

Defluviicoccus 0 0 0 0 45 0 1 0

Dehalococcoides 0 3 0 0 0 0 0 0

Dehalogenimonas 3 1 0 0 0 0 0 0

Deinococcus 0 0 5 4 62 465 64 1184

Delftia 78 60 3047 2221 185 28 172 13

Desulfobacca 3 1 0 0 0 0 0 0

Treatments (mg Cl-/L) Genera 3 50 300 2500 Desulfobulbus 0 0 29 0 67 40 14 4

Desulfocurvus 5 0 77 0 0 169 0 3

Desulforegula 6 0 0 51 0 22 0 0

Desulfotalea 0 3 0 0 0 0 0 0

Desulfurobacterium 0 0 1 0 162 0 0 0

Desulfuromusa 4 0 0 0 1 1 1 39

Devosia 513 602 337 852 294 314 506 307

Diaphorobacter 14 42 0 0 0 0 0 0

Diplorickettsia 2 5 0 0 0 0 0 0

Dongia 6 0 0 0 0 0 0 0

Duganella 587 322 64 283 2 1 38 48

Dyadobacter 1 21 45 113 0 2 77 1

Dyella 5 5 0 0 0 0 0 0

Ectothiorhodospira 3 0 0 0 0 0 0 0

Ectothiorhodosinus 0 0 36 0 0 25 0 0

Ehrlichia 0 0 24 0 0 0 0 0

Elstera 5 6 0 0 0 0 0 0

Emticicia 124 397 886 813 814 12 86 3

Enhydrobacter 0 0 1 2 338 2 0 0

Ensifer 52 256 0 0 0 15 0 0

Enterococcus 1 2 0 34 0 0 0 0

Epulopiscium 5 0 0 0 0 0 0 0

Erythrobacter 14 5 45 245 0 1 3 2

Estrella 0 0 31 0 0 0 0 0

Eucalyptus 0 0 0 0 0 2 3 1

Exiguobacterium 14 19 37 1 45 1 0 42

Ferribacterium 0 0 0 78 0 0 1 0

Ferrimicrobium 0 3 0 0 0 0 0 0

Ferrithrix 3 5 0 0 35 0 81 1

Ferruginibacter 951 1111 3174 5256 2511 1825 109 18

Fibrella 5 3 0 0 0 0 0 0

Fibrobacter 6 0 0 0 0 0 0 0

Filimonas 18 32 248 11 1 3 0 0

Fimbriimonas 0 18 0 0 0 0 53 0

Finegoldia 0 0 0 0 59 0 0 2

Flavihumibacter 42 99 123 190 118 236 0 0

Flavisolibacter 0 3 97 11 1328 4 5 5

Treatments (mg Cl-/L) Genera 3 50 300 2500 Flavobacterium 28701 31962 17358 18024 13141 23286 23212 11212

Flectobacillus 5 30 0 0 0 0 0 0

Flexibacter 26 50 0 161 1 0 98 0

Fluviicola 32 24 0 0 0 28 103 1

Fluviimonas 16 6 0 0 0 0 0 0

Formivibrio 55 86 0 0 0 0 0 0

Frankia 0 3 0 0 11 0 0 0

Frateuria 0 0 0 0 86 0 0 0

Friedmanniella 42 14 40 0 36 267 0 2

Frondihabitans 0 0 0 0 0 1 0 0

Fusobacterium 4 0 0 0 0 0 0 0

Gaiella 13 8 176 81 21 2 1 1

Gallionella 0 4 0 0 0 0 0 0

Gemella 0 0 0 53 40 0 0 0

Gemmata 47 26 135 120 216 93 0 1

Gemmatimonas 48 14 0 101 120 0 0 1

Gemmobacter 8 11 28 51 44 3 14 5

Geoalkalibacter 0 0 78 2 2 1 1 1

Geobacillus 0 7 0 0 0 0 0 0

Geobacter 77 60 60 0 0 0 70 16

Geodermatophilus 15 58 0 0 1 0 18 0

Georgenia 0 0 0 34 52 0 1 0

Georgfuchsia 19 33 29 2 1 14 1 1

Geothermobacter 4 0 0 0 0 0 34 0

Globicatella 0 0 0 0 0 0 3 0

Gordonibacter 0 4 0 0 0 0 0 0

Haematobacter 0 10 0 0 0 0 0 0

Haliangium 1 15 0 0 0 0 0 0

Haemophilus 0 0 0 3 1 0 0 211

Haliscomenobacter 150 209 521 886 475 230 3 3

Halochromatium 0 0 0 0 107 17 0 0

Halomonas 0 0 0 74 28 1 0 37

Halospirulina 1833 1268 309 5 198 507 2325 285

Helicobacter 6 16 0 0 0 0 0 0

Heliospora 0 0 0 0 0 0 3 0

Herbaspirillum 11 72 96 289 156 75 84 6

Herminiimonas 9 2 0 0 0 0 0 0

Treatments (mg Cl-/L) Genera 3 50 300 2500 Herpetosiphon 34 125 1 1 252 1 211 0

Holophaga 4 3 26 114 0 0 0 0

Humitalea 0 3 0 0 0 0 0 0

Hydrogenophaga 474 650 970 1039 885 236 1128 408

Hymenobacter 2 7 1 0 449 11 0 26

Hyphomicrobium 90 70 170 245 37 79 2 19

Hyphomonas 66 71 181 83 128 162 127 63

Iamia 3 5 0 12 47 1 0 0

Ideonella 105 196 56 18 227 39 35 12

Ignavibacterium 0 0 0 115 0 0 0 0

Ilumatobacter 45 55 222 285 3 89 33 47

Inhella 0 12 70 65 0 0 0 0

Iodobacter 0 0 0 0 79 1 98 53

Isoptericola 0 0 0 0 33 0 0 0

Isosphaera 0 0 36 1 0 0 0 0

Kaistia 3 3 0 0 0 0 0 0

Ketogulonicigenium 0 0 0 0 0 1 0 41

Kineococcus 38 28 1 0 0 20 0 31

Kineosporia 35 740 227 93 3 60 37 2

Kitasatospora 2 1 0 0 0 0 0 0

Klebsiella 0 0 58 344 2193 86 576 142

Kocuria 8 1 0 0 3 0 0 0

Labrys 4 1 0 0 0 0 0 0

Lachnoclostridium 4 1 0 0 0 0 0 0

Lacibacter 2 12 0 2 7 0 119 0

Lacibacterium 26 17 0 0 0 0 0 0

Lactobacillus 73 54 414 325 18546 6276 20916 10217

Lampropedia 0 0 1 0 0 1 85 0

Leadbetterella 455 620 570 776 677 461 609 317

Leeuwenhoekiella 0 0 1 0 93 0 0 0

Legionella 360 288 311 428 311 416 7 29

Leptolyngbya 26 6 0 0 0 0 0 0

Leptothrix 95 224 363 353 215 32 4 4

Leucobacter 0 0 1 2 207 84 29 12

Levilinea 0 4 0 0 0 0 0 0

Lewinella 36 34 203 247 225 56 1 0

Limnohabitans 0 0 0 1 2 67 0 0

Treatments (mg Cl-/L) Genera 3 50 300 2500 Loktanella 0 0 0 0 2 1 0 0

Longilinea 3 13 54 0 1 0 0 23

Luedemannella 1 10 0 0 0 0 0 0

Luteococcus 0 0 0 0 0 2 13 0

Luteolibacter 173 425 5384 6903 565 1781 3438 124

Lysinibacillus 2 7 0 1 152 0 0 0

Lysinimicrobium 53 36 63 0 0 0 1 0

Lysobacter 52 38 0 1 45 123 0 2

Magnetospirillum 0 5 0 0 0 0 0 0

Marinobacter 2 3 0 0 0 0 0 0

Marinithermus 0 0 0 0 0 0 0 46

Marmoricola 4 0 29 44 23 0 0 0

Massilia 287 1696 4173 4508 3452 873 2780 823

Meiothermus 0 0 0 0 0 0 0 57

Mesorhizobium 28 38 45 5 112 42 1 29

Metallibacterium 0 6 0 0 0 0 0 0

Methanosaeta 0 0 4 0 57 0 0 0

Methanosphaera 0 0 9 0 0 0 0 0

Methylibium 2203 2309 3266 2781 3084 990 1092 841

Methylobacillus 11 15 0 0 0 0 0 0

Methylobacterium 609 778 825 419 173 580 128 34

Methylocystis 16 21 28 0 17 1 34 29

Methyloferula 79 14 1 76 0 18 1 59

Methyloligella 0 0 0 1 1 0 0 0

Methylomicrobium 0 0 49 0 0 0 0 0

Methylomonas 6 9 0 0 0 0 0 0

Methylophilus 129 167 333 83 99 221 274 44

Methylosinus 0 3 102 0 0 0 1 1

Methylotenera 1253 1445 1297 2172 567 957 229 242

Methylovorus 3 0 0 0 0 0 0 0

Micavibrio 31 23 0 0 0 0 0 0

Microbacterium 24 17 85 5 39 15 37 77

Microbulbifer 0 0 0 0 0 14 0 0

Micrococcus 0 0 2 0 2 0 0 2

Microlunatus 1 4 0 0 29 27 0 0

Microvirga 0 0 53 1 24 0 0 1

Modestobacter 2 1 0 0 0 0 0 0

Treatments (mg Cl-/L) Genera 3 50 300 2500 Morus 21 0 0 0 0 0 0 0

Mucilaginibacter 0 0 1 1 0 0 0 0

Mycobacterium 154 55 523 158 302 83 51 70

Myxozyma 4 4 0 0 0 0 0 0

Nakamurella 11 8 0 0 0 0 0 0

Nannocystis 0 0 0 87 0 0 2 0

Neochlamydia 10 1 0 0 31 1 0 54

Niastella 61 72 518 273 79 100 4 3

Nitratireductor 5 14 0 0 0 0 0 0

Nitrosococcus 28 15 0 68 99 116 3 0

Nitrososphaera 0 0 0 0 0 0 36 0

Nitrosospira 0 0 0 0 133 2 0 0

Nitrosovibrio 6 0 118 1 157 0 0 0

Nitrospira 1 8 0 0 0 0 0 0

Nitrospina 0 0 0 0 0 0 0 1

Nitrospirillum 0 0 0 0 0 18 0 0

Nocardioides 16 22 30 66 444 2 66 44

Nocardiopsis 4 4 0 0 0 0 0 0

Nonomuraea 0 0 0 33 1 1 1 0

Nordella 34 19 105 67 1 2 62 1

Novispirillum 9 6 0 0 0 0 0 0

Novosphingobium 758 1216 751 1510 391 473 1842 595

Ochrobactrum 3 5 0 0 0 0 0 0

Odoribacter 0 0 0 0 32 0 0 0

Oenothera 130 77 0 49 93 37 129 23

Ohtaekwangia 19 48 131 76 6 2 1 0

Opitutus 2 18 0 0 0 0 0 0

Olsenella 0 0 0 0 0 0 0 34

Owenweeksia 15 8 134 89 0 0 1 0

Oxalobacter 4 3 0 0 0 0 0 0

Paenibacillus 61 92 1 57 0 108 26 1

Paludibacter 48 79 0 0 0 0 0 0

Paludibacterium 6 6 0 0 0 0 0 0

Pantoea 268 197 0 0 0 0 0 0

Parachlamydia 4 3 86 0 0 1 0 0

Paracoccus 17 8 1 2 504 2 64 22

Paracraurococcus 4 1 0 0 0 0 0 0

Treatments (mg Cl-/L) Genera 3 50 300 2500 Parasegetibacter 4 18 141 0 2 55 41 3

Parvularcula 0 3 0 0 0 0 0 0

Parvibaculum 0 0 0 0 0 0 9 1

Pasteuria 0 0 69 46 0 0 0 0

Paucibacter 220 807 393 457 287 414 23 56

Paucimonas 0 0 0 0 0 39 0 15

Pectobacterium 116 149 0 0 28 38 0 0

Pedobacter 43 399 308 88 138 1 626 3

Pedomicrobium 3 1 0 0 0 0 0 0

Pelobacter 3 0 0 0 0 0 0 0

Pedosphaera 0 0 58 93 1 1 1 0

Pelomonas 98 245 224 115 43 64 59 32

Pelosinus 8 7 127 1 0 0 0 1

Pelotomaculum 18 13 9 331 0 145 2 0

Peptoniphilus 0 0 0 0 42 0 0 0

Peptostreptococcus 0 0 0 0 18 0 0 0

Peredibacter 16 0 0 0 148 0 0 0

Phenylobacterium 108 97 2 43 93 110 1 51

Phormidium 23 1 0 0 0 0 0 0

Phycisphaera 68 37 170 102 330 81 0 2

Phyllobacterium 2 3 0 0 0 0 0 0

Phytophthora 4 6 15 7 15 5 1 0

Picochlorum 0 5 0 0 0 0 0 0

Pirellula 151 91 411 461 174 200 83 174

Planctomyces 63 29 122 168 252 101 49 17

Planomicrobium 4 3 0 0 0 0 0 0

Pleomorphomonas 4 5 0 0 0 0 0 0

Planococcus 0 0 0 0 0 0 0 1

Pleomorphobacterium 0 0 0 0 1 1 0 0

Polaromonas 132 97 5 1 150 161 99 18

Polyangium 0 0 0 0 0 51 0 0

Porphyrobacter 54 27 26 113 44 1 37 96

Prevotella 0 0 1 78 61 0 0 15

Prochlorococcus 7 4 0 21 0 0 0 0

Propionibacterium 15 7 43 170 411 48 39 98

Propionicimonas 0 3 0 0 0 23 0 0

Propionigenium 4 1 0 0 0 0 0 0

Treatments (mg Cl-/L) Genera 3 50 300 2500 Prosthecobacter 8 26 177 408 1 1 1 0

Prototheca 0 3 0 0 0 0 0 0

Pseudolabrys 5 0 0 0 0 0 0 0

Proteus 0 0 0 0 0 4 1 0

Pseudoduganella 0 0 3 124 1 0 0 1

Pseudomonas 2711 4404 1647 1431 1375 881 5815 716

Pseudonocardia 0 8 0 1 240 40 2 2

Pseudospirillum 0 0 1 0 0 1 0 24

Pseudoxanthomonas 32 6 0 0 1 0 0 0

Quadrisphaera 18 45 11 0 0 0 0 0

Quatrionicoccus 1 5 0 0 0 0 0 0

Rahnella 113 321 0 0 0 0 0 0

Ralstonia 6 9 4 2 834 3 1 2

Ramlibacter 0 0 41 1 1 12 0 0

Reyranella 7 3 0 1 61 0 0 0

Rheinheimera 3 21 0 0 0 0 0 0

Rhizobacter 1054 1327 0 0 0 0 0 0

Rhizobium 10021 9194 5373 5695 9033 2171 2711 828

Rhizomicrobium 7 1 0 0 0 0 0 0

Rhodanobacter 0 0 1 1 3 0 0 0

Rhodobacter 1600 1162 2417 2691 898 1607 1823 393

Rhodobium 9 2 58 0 1 0 35 1

Rhodocista 48 60 0 0 128 0 0 0

Rhodococcus 64 34 1 1 196 0 66 0

Rhodocyclus 41 60 0 0 0 0 0 0

Rhodoferax 1499 1559 420 893 396 88 61 62

Rhodomicrobium 191 184 107 53 2 3 41 0

Rhodopirellula 0 3 0 0 0 0 0 27

Rhodoplanes 4 10 77 41 0 0 186 1

Rhodopseudomonas 5 4 0 0 0 0 0 0

Rhodospirillum 20 67 117 0 0 0 1 1

Rhodovastum 7 0 0 0 0 0 0 0

Rhodovulum 4 7 1 0 0 0 0 0

Rickettsia 25 61 6 54 0 0 0 0

Rickettsiella 0 0 0 46 0 0 0 1

Roseateles 171 230 2 248 11 2 50 25

Roseibacillus 7 19 80 57 84 105 58 11

Treatments (mg Cl-/L) Genera 3 50 300 2500 Roseibacterium 0 0 58 1 0 0 0 0

Roseiflexus 0 10 0 0 0 0 5 0

Roseimaritima 0 3 0 0 0 0 0 0

Roseobacter 3 0 0 0 0 0 0 0

Roseococcus 5 1 78 0 0 0 0 0

Roseomonas 51 55 50 103 52 42 62 1

Rubellimicrobium 0 0 0 0 132 0 0 0

Rubrivivax 155 226 204 215 272 154 18 57

Rubrobacter 0 0 36 0 196 1 0 1

Runella 36 62 267 121 282 2 2 1

Saccharibacillus 8 2 0 0 0 0 0 0

Saccharibacter 0 6 0 0 0 0 32 0

Saccharospirillum 4 9 0 0 0 0 0 0

Saccharopolyspora 0 0 0 1 0 2 0 0

Sandaracinus 5 3 5 0 0 0 0 0

Sandarakinorhabdus 0 8 0 1 82 0 0 0

Sanguibacter 142 127 0 0 0 0 0 0

Schlegelella 0 0 0 0 0 0 43 0

Sediminibacterium 63 208 105 108 2 2 1 1

Sideroxydans 0 4 0 0 0 0 0 0

Simaetha 0 3 0 0 0 0 0 0

Serratia 0 0 0 0 0 1 0 26

Shigella 0 0 1 2 239 47 413 143

Simplicispira 9 29 17 4 6 0 21 1

Sinorhizobium 3 11 0 0 0 0 0 0

Smithella 0 3 0 0 0 0 0 0

Solibacillus 0 5 0 0 0 0 0 0

Singulisphaera 0 0 0 0 44 0 0 0

Solirubrobacter 0 0 0 0 0 0 5 0

Sorangium 10 0 0 0 0 0 48 0

Sphaerobacter 0 0 109 0 47 1 0 1

Sphaerotilus 365 290 373 570 453 126 4 71

Sphingobacterium 41 63 310 270 59 2 2 3

Sphingobium 356 723 145 373 8 158 1144 417

Sphingomonas 453 512 473 105 814 86 132 166

Sphingopyxis 757 591 1025 1348 937 666 660 398

Sphingorhabdus 24 20 156 34 87 94 44 86

Treatments (mg Cl-/L) Genera 3 50 300 2500 Spirosoma 11 0 0 0 149 18 0 0

Sporichthya 13 4 0 0 0 1 0 41

Sporobacter 0 6 0 0 0 0 0 0

Staphylococcus 13 10 105 105 1041 5 3 9

Starkeya 14 3 2 3 3 0 0 0

Stenotrophomonas 477 275 242 178 28 212 3 2

Steroidobacter 19 30 67 3 0 44 1 0

Sterolibacterium 198 181 217 39 1 86 0 0

Streptococcus 0 0 1 621 1429 1 90 90

Streptomyces 0 0 1 94 125 3 1 2

Sulfurospirillum 4 1 0 0 0 0 0 0

Sulfitobacter 0 0 0 0 1 0 0 2

Sulfurimonas 0 0 0 1 67 1 1 1

Sulfurovum 0 0 17 1 106 151 0 1

Synechococcus 0 0 0 0 0 0 0 4

Syntrophus 0 0 133 0 0 0 0 1

Tabrizicola 361 539 0 0 0 0 0 0

Tepidamorphus 0 0 0 0 54 0 0 1

Tepidimicrobium 0 0 0 0 0 0 0 1

Tepidimonas 0 0 0 73 3 0 1 1

Terrabacter 0 4 0 0 0 0 0 0

Terrimonas 39 85 666 655 488 3 18 2

Thalassospira 11 4 0 0 0 0 0 0

Thauera 3 3 0 0 0 0 0 0

Thermanaerothrix 21 19 0 0 0 0 0 0

Tessaracoccus 0 0 0 0 2 1 1 0

Tetrasphaera 0 0 0 0 58 0 1 22

Thermicanus 0 3 1 0 0 0 0 0

Thermoflavimicrobium 0 0 81 0 0 1 0 0

Thermomonas 98 270 86 136 73 145 1 2

Thermovum 2 1 0 0 0 0 0 0

Thermus 0 3 33 0 0 0 0 0

Thioalkalivibrio 0 3 0 0 1 31 0 0

Thiohalophilus 0 0 0 0 69 1 0 1

Thiomicrospira 0 0 0 0 0 7 0 0

Thioprofundum 0 3 0 0 0 1 0 4

Thiorhodospira 4 2 0 0 0 0 0 0

Treatments (mg Cl-/L) Genera 3 50 300 2500 Thiothrix 0 0 0 185 1 2 0 1

Tissierella 0 0 49 0 0 0 0 1

Tolumonas 1185 1397 94 66 0 1 0 1

Trichococcus 62 247 205 844 124 95 213 63

Turicibacter 0 0 0 2 0 0 0 1

Uliginosibacterium 3404 2349 258 145 25 142 18 7

Undibacterium 21 22 68 0 40 73 1 1

Variovorax 72 100 469 311 432 131 349 48

Veillonella 0 0 0 84 0 0 1 40

Verrucomicrobium 31 54 549 391 3 297 0 0

Vibrio 11 0 0 0 0 0 0 0

Williamsia 0 0 0 0 26 0 0 0

Wolbachia 0 0 200 29 0 1 0 0

Woodsholea 0 0 0 1 51 1 0 0

Xanthomonas 1 24 171 0 0 15 0 1

Xylella 11 5 0 0 0 0 0 0

Yonghaparkia 199 98 0 0 0 0 0 0

Zavarzinella 1 7 0 0 0 0 0 39

Zea 0 0 0 43 0 0 0 0

Zebrasoma 0 5 0 0 0 0 0 0

Zoogloea 17 43 0 0 0 0 0 0 total counts 90860 99848 86494 91594 98334 125996 88497 101648

APPENDIX C

Genera bacteria relative abundance (percentage) data for all treatments (Two replicates are indicated per treatment)

Treatments (mg Cl-/L) Genera 3 50 300 2500 Acanthamoeba 0 0.0160 0 0 0 0 0 0

Acetivibrio 0 0.0060 0 0 0 0 0 0

Acetobacterium 0.0187 0.0751 0 0 0 0 0 0

Acidimicrobium 0.0110 0.0030 0 0 0 0 0 0

Acidisphaera 0.0220 0.0070 0 0 0 0 0 0

Aciditerrimonas 0.0286 0.0431 0 0 0 0 0 0

Acidiferrobacter 0 0 0 0 0 0 0.0124 0

Aciditerrimonas 0 0 0 0 0.1434 0.0008 0.0802 0.0502

Acidobacterium 0.0407 0.0220 0.4601 0.0917 0.1617 0.1318 0 0.0020

Acidothermus 0 0 0.0012 0 0 0.0016 0 0

Acidovorax 4.9120 6.4929 4.7182 2.6912 1.1583 0.5413 2.2588 1.1894

Acinetobacter 0.0374 0.0180 0.3353 0.3570 0.4708 0.0468 0.9582 0.0295

Actinomadura 0 0.0040 0.0012 0 0.3529 0.0008 0 0.0010

Actinomyces 0 0 0.0023 0.0317 0.0651 0.0024 0 0

Actinomycetospora 0.0044 0.0050 0 0 0.0203 0.0540 0 0

Actinoplanes 0.2014 0.4687 1.3677 0.4345 0.1780 0.0317 0.8633 0.0128

Adhaeribacter 0.0044 0 0 0 0 0 0 0

Aerococcus 0 0 0.0012 0 0 0.0857 0 0

Aeromonas 1.0489 0.4256 0.6613 0.3101 0.4271 50.5310 0.6204 62.6780

Agrobacterium 0.8155 0.7111 0.7989 0.7500 0.5390 0.2326 0.2870 0.0580

Agrococcus 0.0880 0.0040 0.1098 0.0011 0.0010 0 0 0

Alcanivorax 0.0055 0 0 0 0 0 0 0

Alkanindiges 0.0044 0.0060 0 0 0 0 0 0

Alloactinosynnema 0.0055 0 0 0 0 0 0 0

Allokutzneria 0 0.0040 0 0 0 0 0 0

Algorimarina 0 0 0 0.1823 0 0 0 0.0010

Altererythrobacter 0.1002 0.1082 0.1387 0.7806 0.0031 0.0817 0.1164 0.0649

Alteromonas 0 0 0 0 0 0 0 0.0010

Amaricoccus 0.0198 0.0611 0.0023 0.0011 0.0864 0 0.1345 0.0285

Aminobacter 0.0121 0.0240 0 0 0 0 0 0

Amnibacterium 0 0 0.1861 0.0830 0.8929 0.3818 0.0090 0.0246

Anaerococcus 0 0 0 0 0.1210 0.0540 0 0

Anaerolinea 0.0209 0.0471 0.0474 0.0677 0 0.0389 0 0.0010

Anaeromyxobacter 0.0176 0.0391 0.0012 0 0.0010 0.0016 0.0147 0

Ancalomicrobium 0 0.0030 0 0 0 0 0 0

Aquabacterium 0.3907 0.2604 0.5249 0.7872 0.0681 0.1056 0.3198 0.0748

Aquicella 0.0605 0.0130 0.1029 0.0011 0.0539 0.0008 0.0023 0

Treatments (mg Cl-/L) Genera 3 50 300 2500 Aquimonas 0.0044 0.0080 0 0.0011 0.0010 0.0198 0.0554 0.0010

Aquincola 0.0022 0.0050 0 0.0022 0.0031 0 0 0

Archaeoglobus 0 0 0.0312 0 0 0 0 0

Arcicella 0.1079 0.2213 0.4786 0.7522 0.1027 0.1778 0.0045 0.0039

Arcobacter 0.0077 0.0461 0 0.1878 0 0.0008 0 0

Arenimonas 0.1640 0.2273 0.4613 0.0229 0.0498 0.0333 0.0023 0.0394

Arthrobacter 0.0275 0.0260 1.7678 1.5536 5.9461 2.6612 4.4013 2.1456

Asticcacaulis 3.0013 3.3110 1.9204 0.9411 0.9366 0.8357 0.7424 0.3276

Atopostipes 0 0 0 0 0 0 0 0.0010

Aurantimonas 0 0.0120 0 0 0 0.0175 0 0

Azoarcus 0.0022 0.0010 0 0 0 0 0 0

Azonexus 0.0121 0.0100 0 0.1321 0 0 0.0011 0

Azorhizobium 0.0099 0.0120 0 0 0.0590 0 0 0

Azospira 0.0099 0.0531 0 0 0 0 0 0

Azospirillum 0.0055 0.0070 0 0 0 0 0 0

Bacillus 0.0726 0.0170 0.0821 0.1703 0.2278 0.1564 0.1232 0.0030

Bacteriovorax 0.0154 0.0471 0.0555 0.0011 0.1180 0 0 0.0010

Bacteroides 0.0154 0.0220 0 0.0011 0.1271 0.0516 0 0

Bdellovibrio 0.0451 0.0451 0 0 0 0 0 0

Beggiatoa 0.0242 0.0290 0 0 0 0 0 0

Beijerinckia 0.1926 0.1492 0.1249 0.3330 0.0488 0 0.0203 0.0020

Bergeyella 0 0 0 0 0.2420 0 0 0.0010

Blastochloris 0.0011 0.0020 0 0 0 0.0254 0 0

Blastococcus 0 0 0.0243 0 0.2512 0.0016 0.0011 0

Blastomonas 0 0 0 0.0011 0 0 0.0870 0

Blastopirellula 0.0176 0.0120 0.2705 0.0328 0.0295 0 0.0011 0.0010

Bosea 0.0969 0.1102 0.0231 0.0600 0.0010 0.0540 0 0.0020

Brachybacterium 0.0077 0 0 0.0011 0 0 0.0011 0.0610

Bradyrhizobium 0.1266 0.1482 0.0012 0.1463 0.0010 0.0016 0 0.0030

Brenneria 0 0.0371 0 0 0 0 0 0

Brevibacillus 0.0044 0 0 0 0 0 0 0.0797

Brevundimonas 2.4037 2.3075 0.8567 0.3494 0.6224 0.5683 0.5141 0.4043

Brooklawnia 0 0 0 0 0 0 0 0.0108

Brucella 0.0660 0.0601 0.0728 0.1441 0.0681 0.0286 0.0011 0

Buttiauxella 1.8952 0.9474 0 0 0 0 0 0

Burkholderia 0.0066 0.0180 0.2451 0.1419 0.1780 0.0595 0.0011 0

Byssovorax 0 0.0030 0 0.0666 0.0915 0 0 0

Treatments (mg Cl-/L) Genera 3 50 300 2500 Bythopirellula 0.0110 0 0.0751 0 0.0010 0 0 0.0010

Caedibacter 0.0451 0.0300 0 0.1004 0.0010 0.0183 0 0

Caldilinea 0 0 0 0.0535 0 0 0 0

Candidatus Acetothermum 0 0 0 0.0011 0 0 0 0

Candidatus Alysiosphaera 0 0 0 0.0011 0.0753 0.0016 0.0305 0

Candidatus Babela 0 0.0040 0 0 0 0 0 0

Candidatus Captivus 0.0033 0 0 0 0 0 0 0

Candidatus Carsonella 0.0033 0.0010 0 0 0 0 0 0

Candidatus Chloracidobacterium 0 0 0 0 0.0397 0.0008 0 0

Candidatus Kuenenia 0 0 0 0 0 0 0 0.0039

Candidatus Lariskella 0.0033 0 0 0 0 0 0 0

Candidatus Metachlamydia 0.0451 0.0551 0.1434 0 0 0.2056 0 0.0010

Candidatus Microthrix 0.0198 0.0010 0.1029 0.0917 0 0.0611 0.0034 0

Candidatus Midichloria 0.0231 0.0280 0.0751 0 0 0 0 0

Candidatus Nitrosoarchaeum 0 0 0.0023 0.0022 0.0031 0 0 0

Candidatus Nitrososphaera 0 0 0 0 0 0.0024 0 0

Candidatus Nucleicultrix 0.0297 0.0280 0 0 0 0 0 0

Candidatus Odyssella 0.0176 0.0310 0.2636 0.0011 0.0010 0.0492 0 0.0020

Candidatus Pelagibacter 0 0 0.0197 0 0.1180 0.0024 0.0011 0.0010

Candidatus Phytoplasma 0.0671 0.0120 0.0035 0.2773 0.8064 0.0944 0.4520 0.0571

Candidatus Planktophila 0 0 0 0 0 0 0 0.0374

Candidatus Protochlamydia 0.0110 0 0 0 0.0905 0 0 0

Candidatus Rhabdochlamydia 0.0088 0.0020 0 0 0 0.0484 0 0

Candidatus Saccharimonas 0 0.0080 0.0254 0.1004 0.2705 0.0325 0.0362 0.0010

Candidatus Solibacter 0 0 0.1538 0 0.0020 0.0008 0 0

Candidatus Trichorickettsia 0.0033 0 0 0 0 0 0 0

Candidatus Vidania 0.0011 0.0020 0.0624 0 0.0010 0.0492 0 0

Candidatus Xiphinematobacter 0 0.0030 0 0 0 0 0 0

Capnocytophaga 0 0 0 0 0 0 0.0169 0

Carboxydothermus 0 0 0 0 0.1281 0 0 0

Carnobacterium 0.0044 0.0010 0 0 0 0 0.0486 0.0010

Catellicoccus 0 0 0 0 0.0010 0.0008 0.1424 0

Catenuloplanes 0 0.0090 0 0 0 0 0 0

Caulobacter 0.6031 0.9775 0.4798 0.3068 0.1892 0.0968 0.1254 0.1230

Cellulomonas 4.7182 1.4953 0.5526 0.7500 0.2827 0.3286 0.1718 0.0689

Cellvibrio 0.1068 0.0511 0.1572 0.0502 0 0.0437 0.0011 0.0413

Chamaesiphon 0.0077 0 0 0 0 0 0 0

Treatments (mg Cl-/L) Genera 3 50 300 2500 Chelatococcus 0.0286 0.0120 0.0058 0.1179 0.0020 0.0024 0.0452 0.0679

Chitinimonas 0.0077 0.0080 0 0 0 0 0 0

Chitinophaga 0.2234 0.5268 0.4151 0.0066 0.0031 0.0556 0 0.0020

Chloroflexus 0.0033 0.0130 0.1411 0.0022 0.0559 0.0016 0.0893 0.0010

Chloroidium 0.0066 0 0 0 0 0 0 0

Chondromyces 0.0066 0.0010 0 0 0 0 0 0

Chroococcidiopsis 0 0 0 0 0.2227 0 0 0

Chryseobacterium 0.0044 0 0.0439 0.0011 0 0 0 0

Chthoniobacter 0.0044 0.0250 0 0.1015 0 0.1230 0.0011 0

Citreicella 0.0693 0.0320 0 0 0 0 0 0

Cloacibacterium 0 0.0050 0.0012 1.7720 0.4546 0.0056 0.0011 0.0020

Clostridium 0.0737 0.0741 0.2185 0.1004 0.2227 0.0056 0.0621 0.0059

Cohnella 0 0.0030 0 0 0 0 0 0

Collimonas 0 0 0.0983 0 0.0010 0 0 0

Collinsella 0 0 0 0 0.0305 0 0 0

Comamonas 0.0220 0.0521 0.0613 0.0928 0 0.0008 0.1322 0.0030

Compostimonas 0.0385 0.0130 0.0012 0 0.0010 0.0508 0 0.0787

Conexibacter 0.0242 0.0270 0.0127 0 0.2563 0.0008 0.0520 0.0512

Corynebacterium 0.0011 0.0150 0.1468 1.0066 0.8390 0.0095 0.0079 0.0305

Coxiella 0.0286 0.0210 0.0023 0.1026 0 0.0587 0.0294 0.0020

Crocinitomix 0.0803 0.0621 0.3596 0.4214 0.0803 0.2548 0 0.0039

Cronobacter 0 0 0 0 0.0417 0 0.0011 0

Cryocola 0 0 0.0069 0.0076 0.4688 0.0079 0.0045 0.0079

Cupriavidus 0.0033 0.0020 0 0 0 0 0 0

Curtobacterium 0.2168 0.0341 0 0 0.0183 0.0770 0.0056 0

Curvibacter 0.0044 0.0020 0.2208 0.0863 0.0508 0.0278 0.0554 0.0079

Cyanidium 0.0165 0.0060 0 0 0 0 0 0

Cystobacter 0.0055 0 0 0 0 0 0 0

Cytophaga 0.1398 0.3305 0.2046 0.3308 0.2105 0.0016 0.0633 0.0020

Daeguia 0 0 0.1075 0.1310 0.0712 0.1302 0.0249 0.0167

Dechloromonas 0.1112 0.1222 0.0798 0.0011 0 0.0389 0 0.0020

Deefgea 0 0.0050 0.0867 0 0 0 0.0011 0

Defluviicoccus 0 0 0 0 0.0458 0 0.0011 0

Dehalococcoides 0 0.0030 0 0 0 0 0 0

Dehalogenimonas 0.0033 0.0010 0 0 0 0 0 0

Deinococcus 0 0 0.0058 0.0044 0.0631 0.3691 0.0723 1.1648

Delftia 0.0858 0.0601 3.5228 2.4248 0.1881 0.0222 0.1944 0.0128

Treatments (mg Cl-/L) Genera 3 50 300 2500 Desulfobacca 0.0033 0.0010 0 0 0 0 0 0

Desulfobulbus 0 0 0.0335 0 0.0681 0.0317 0.0158 0.0039

Desulfocurvus 0.0055 0 0.0890 0 0 0.1341 0 0.0030

Desulforegula 0.0066 0 0 0.0557 0 0.0175 0 0

Desulfotalea 0 0.0030 0 0 0 0 0 0

Desulfurobacterium 0 0 0.0012 0 0.1647 0 0 0

Desulfuromusa 0.0044 0 0 0 0.0010 0.0008 0.0011 0.0384

Devosia 0.5646 0.6029 0.3896 0.9302 0.2990 0.2492 0.5718 0.3020

Diaphorobacter 0.0154 0.0421 0 0 0 0 0 0

Diplorickettsia 0.0022 0.0050 0 0 0 0 0 0

Dongia 0.0066 0 0 0 0 0 0 0

Duganella 0.6460 0.3225 0.0740 0.3090 0.0020 0.0008 0.0429 0.0472

Dyadobacter 0.0011 0.0210 0.0520 0.1234 0 0.0016 0.0870 0.0010

Dyella 0.0055 0.0050 0 0 0 0 0 0

Ectothiorhodosinus 0 0 0.0416 0 0 0.0198 0 0

Ectothiorhodospira 0.0033 0 0 0 0 0 0 0

Ehrlichia 0 0 0.0277 0 0 0 0 0

Elstera 0.0055 0.0060 0 0 0 0 0 0

Emticicia 0.1365 0.3976 1.0243 0.8876 0.8278 0.0095 0.0972 0.0030

Enhydrobacter 0 0 0.0012 0.0022 0.3437 0.0016 0 0

Ensifer 0.0572 0.2564 0 0 0 0.0119 0 0

Enterococcus 0.0011 0.0020 0 0.0371 0 0 0 0

Epulopiscium 0.0055 0 0 0 0 0 0 0

Erythrobacter 0.0154 0.0050 0.0520 0.2675 0 0.0008 0.0034 0.0020

Estrella 0 0 0.0358 0 0 0 0 0

Eucalyptus 0 0 0 0 0 0.0016 0.0034 0.0010

Exiguobacterium 0.0154 0.0190 0.0428 0.0011 0.0458 0.0008 0 0.0413

Ferribacterium 0 0 0 0.0852 0 0 0.0011 0

Ferrimicrobium 0 0.0030 0 0 0 0 0 0

Ferrithrix 0.0033 0.0050 0 0 0.0356 0 0.0915 0.0010

Ferruginibacter 1.0467 1.1127 3.6696 5.7384 2.5535 1.4485 0.1232 0.0177

Fibrella 0.0055 0.0030 0 0 0 0 0 0

Fibrobacter 0.0066 0 0 0 0 0 0 0

Filimonas 0.0198 0.0320 0.2867 0.0120 0.0010 0.0024 0 0

Fimbriimonas 0 0.0180 0 0 0 0 0.0599 0

Finegoldia 0 0 0 0 0.0600 0 0 0.0020

Flavihumibacter 0.0462 0.0992 0.1422 0.2074 0.1200 0.1873 0 0

Treatments (mg Cl-/L) Genera 3 50 300 2500 Flavisolibacter 0 0.0030 0.1121 0.0120 1.3505 0.0032 0.0056 0.0049

Flavobacterium 31.588 32.011 20.068 19.678 13.364 18.482 26.2291 11.0302

Flectobacillus 0.0055 0.0300 0 0 0 0 0 0

Flexibacter 0.0286 0.0501 0 0.1758 0.0010 0 0.1107 0

Fluviicola 0.0352 0.0240 0 0 0 0.0222 0.1164 0.0010

Fluviimonas 0.0176 0.0060 0 0 0 0 0 0

Formivibrio 0.0605 0.0861 0 0 0 0 0 0

Frankia 0 0.0030 0 0 0.0112 0 0 0

Frateuria 0 0 0 0 0.0875 0 0 0

Friedmanniella 0.0462 0.0140 0.0462 0 0.0366 0.2119 0 0.0020

Frondihabitans 0 0 0 0 0 0.0008 0 0

Fusobacterium 0.0044 0 0 0 0 0 0 0

Gaiella 0.0143 0.0080 0.2035 0.0884 0.0214 0.0016 0.0011 0.0010

Gallionella 0 0.0040 0 0 0 0 0 0

Gemella 0 0 0 0.0579 0.0407 0 0 0

Gemmata 0.0517 0.0260 0.1561 0.1310 0.2197 0.0738 0 0.0010

Gemmatimonas 0.0528 0.0140 0 0.1103 0.1220 0 0 0.0010

Gemmobacter 0.0088 0.0110 0.0324 0.0557 0.0447 0.0024 0.0158 0.0049

Geoalkalibacter 0 0 0.0902 0.0022 0.0020 0.0008 0.0011 0.0010

Geobacillus 0 0.0070 0 0 0 0 0 0

Geobacter 0.0847 0.0601 0.0694 0 0 0 0.0791 0.0157

Geodermatophilus 0.0165 0.0581 0 0 0.0010 0 0.0203 0

Georgenia 0 0 0 0.0371 0.0529 0 0.0011 0

Georgfuchsia 0.0209 0.0331 0.0335 0.0022 0.0010 0.0111 0.0011 0.0010

Geothermobacter 0.0044 0 0 0 0 0 0.0384 0

Globicatella 0 0 0 0 0 0 0.0034 0

Gordonibacter 0 0.0040 0 0 0 0 0 0

Haematobacter 0 0.0100 0 0 0 0 0 0

Haemophilus 0 0 0 0.0033 0.0010 0 0 0.2076

Haliangium 0.0011 0.0150 0 0 0 0 0 0

Haliscomenobacter 0.1650 0.2093 0.6024 0.9673 0.4830 0.1825 0.0034 0.0030

Halochromatium 0 0 0 0 0.1088 0.0135 0 0

Halomonas 0 0 0 0.0808 0.0285 0.0008 0 0.0364

Halospirulina 2.0174 1.2699 0.3573 0.0055 0.2014 0.4024 2.6272 0.2804

Helicobacter 0.0066 0.0160 0 0 0 0 0 0

Heliospora 0 0 0 0 0 0 0.0034 0

Herbaspirillum 0.0121 0.0721 0.1110 0.3155 0.1586 0.0595 0.0949 0.0059

Treatments (mg Cl-/L) Genera 3 50 300 2500 Herminiimonas 0.0099 0.0020 0 0 0 0 0 0

Herpetosiphon 0.0374 0.1252 0.0012 0.0011 0.2563 0.0008 0.2384 0

Holophaga 0.0044 0.0030 0.0301 0.1245 0 0 0 0

Humitalea 0 0.0030 0 0 0 0 0 0

Hydrogenophaga 0.5217 0.6510 1.1215 1.1344 0.9000 0.1873 1.2746 0.4014

Hymenobacter 0.0022 0.0070 0.0012 0 0.4566 0.0087 0 0.0256

Hyphomicrobium 0.0991 0.0701 0.1965 0.2675 0.0376 0.0627 0.0023 0.0187

Hyphomonas 0.0726 0.0711 0.2093 0.0906 0.1302 0.1286 0.1435 0.0620

Iamia 0.0033 0.0050 0 0.0131 0.0478 0.0008 0 0

Ideonella 0.1156 0.1963 0.0647 0.0197 0.2308 0.0310 0.0395 0.0118

Ignavibacterium 0 0 0 0.1256 0 0 0 0

Ilumatobacter 0.0495 0.0551 0.2567 0.3112 0.0031 0.0706 0.0373 0.0462

Inhella 0 0.0120 0.0809 0.0710 0 0 0 0

Iodobacter 0 0 0 0 0.0803 0.0008 0.1107 0.0521

Isoptericola 0 0 0 0 0.0336 0 0 0

Isosphaera 0 0 0.0416 0.0011 0 0 0 0

Kaistia 0.0033 0.0030 0 0 0 0 0 0

Ketogulonicigenium 0 0 0 0 0 0.0008 0 0.0403

Kineococcus 0.0418 0.0280 0.0012 0 0 0.0159 0 0.0305

Kineosporia 0.0385 0.7411 0.2624 0.1015 0.0031 0.0476 0.0418 0.0020

Kitasatospora 0.0022 0.0010 0 0 0 0 0 0

Klebsiella 0 0 0.0671 0.3756 2.2302 0.0683 0.6509 0.1397

Kocuria 0.0088 0.0010 0 0 0.0031 0 0 0

Labrys 0.0044 0.0010 0 0 0 0 0 0

Lachnoclostridium 0.0044 0.0010 0 0 0 0 0 0

Lacibacter 0.0022 0.0120 0 0 0 0 0 0

Lactobacillus 0.0803 0.0541 0.4786 0.3548 18.8602 4.9811 23.6347 10.0514

Lacibacterium 0.0286 0.0170 0 0 0 0 0 0

Lampropedia 0 0 0.0012 0 0 0.0008 0.0960 0

Leadbetterella 0.5008 0.6209 0.6590 0.8472 0.6885 0.3659 0.6882 0.3119

Leeuwenhoekiella 0 0 0.0012 0 0.0946 0 0 0

Legionella 0.3962 0.2884 0.3596 0.4673 0.3163 0.3302 0.0079 0.0285

Leptothrix 0.1046 0.2243 0.4197 0.3854 0.2186 0.0254 0.0045 0.0039

Leptolyngbya 0.0286 0.0060 0 0 0 0 0 0

Leucobacter 0 0 0.0012 0.0022 0.2105 0.0667 0.0328 0.0118

Levilinea 0 0.0040 0 0 0 0 0 0

Lewinella 0.0396 0.0341 0.2347 0.2697 0.2288 0.0444 0.0011 0

Treatments (mg Cl-/L) Genera 3 50 300 2500 Limnohabitans 0 0 0 0.0011 0.0020 0.0532 0 0

Loktanella 0 0 0 0 0.0020 0.0008 0 0

Longilinea 0.0033 0.0130 0.0624 0 0.0010 0 0 0.0226

Luedemannella 0.0011 0.0100 0 0 0 0 0 0

Luteococcus 0 0 0 0 0 0.0016 0.0147 0

Luteolibacter 0.1904 0.4256 6.2247 7.5365 0.5746 1.4135 3.8849 0.1220

Lysinibacillus 0.0022 0.0070 0 0.0011 0.1546 0 0 0

Lysinimicrobium 0.0583 0.0361 0.0728 0 0 0 0.0011 0

Lysobacter 0.0572 0.0381 0 0.0011 0.0458 0.0976 0 0.0020

Magnetospirillum 0 0.0050 0 0 0 0 0 0

Marinobacter 0.0022 0.0030 0 0 0 0 0 0

Marinithermus 0 0 0 0 0 0 0 0.0453

Marmoricola 0.0044 0 0.0335 0.0480 0.0234 0 0 0

Massilia 0.3159 1.6986 4.8246 4.9217 3.5105 0.6929 3.1413 0.8097

Meiothermus 0 0 0 0 0 0 0 0.0561

Mesorhizobium 0.0308 0.0381 0.0520 0.0055 0.1139 0.0333 0.0011 0.0285

Metallibacterium 0 0.0060 0 0 0 0 0 0

Methanosaeta 0 0 0.0046 0 0.0580 0 0 0

Methanosphaera 0 0 0.0104 0 0 0 0 0

Methylibium 2.4246 2.3125 3.7760 3.0362 3.1362 0.7857 1.2339 0.8274

Methylobacillus 0.0121 0.0150 0 0 0 0 0 0

Methylobacterium 0.6703 0.7792 0.9538 0.4575 0.1759 0.4603 0.1446 0.0334

Methylocystis 0.0176 0.0210 0.0324 0 0.0173 0.0008 0.0384 0.0285

Methyloferula 0.0869 0.0140 0.0012 0.0830 0 0.0143 0.0011 0.0580

Methyloligella 0 0 0 0.0011 0.0010 0 0 0

Methylomicrobium 0 0 0.0567 0 0 0 0 0

Methylomonas 0.0066 0.0090 0 0 0 0 0 0

Methylophilus 0.1420 0.1673 0.3850 0.0906 0.1007 0.1754 0.3096 0.0433

Methylosinus 0 0.0030 0.1179 0 0 0 0.0011 0.0010

Methylotenera 1.3790 1.4472 1.4995 2.3713 0.5766 0.7595 0.2588 0.2381

Methylovorus 0.0033 0 0 0 0 0 0 0

Micavibrio 0.0341 0.0230 0 0 0 0 0 0

Microbacterium 0.0264 0.0170 0.0983 0.0055 0.0397 0.0119 0.0418 0.0758

Microbulbifer 0 0 0 0 0 0.0111 0 0

Micrococcus 0 0 0.0023 0 0.0020 0 0 0.0020

Microlunatus 0.0011 0.0040 0 0 0.0295 0.0214 0 0

Microvirga 0 0 0.0613 0.0011 0.0244 0 0 0.0010

Treatments (mg Cl-/L) Genera 3 50 300 2500 Modestobacter 0.0022 0.0010 0 0 0 0 0 0

Morus 0.0231 0 0 0 0 0 0 0

Mucilaginibacter 0 0 0.0012 0.0011 0 0 0 0

Mycobacterium 0.1695 0.0551 0.6047 0.1725 0.3071 0.0659 0.0576 0.0689

Myxozyma 0.0044 0.0040 0 0 0 0 0 0

Nakamurella 0.0121 0.0080 0 0 0 0 0 0

Nannocystis 0 0 0 0.0950 0 0 0.0023 0

Neochlamydia 0.0110 0.0010 0 0 0.0315 0.0008 0 0.0531

Niastella 0.0671 0.0721 0.5989 0.2981 0.0803 0.0794 0.0045 0.0030

Nitratireductor 0.0055 0.0140 0 0 0 0 0 0

Nitrosococcus 0.0308 0.0150 0 0.0742 0.1007 0.0921 0.0034 0

Nitrososphaera 0 0 0 0 0 0 0.0407 0

Nitrosospira 0 0 0 0 0.1353 0.0016 0 0

Nitrosovibrio 0.0066 0 0.1364 0.0011 0.1597 0 0 0

Nitrospina 0.0011 0.0080 0 0 0 0 0 0.0010

Nitrospirillum 0 0 0 0 0 0.0143 0 0

Nocardioides 0.0176 0.0220 0.0347 0.0721 0.4515 0.0016 0.0746 0.0433

Nocardiopsis 0.0044 0.0040 0 0 0 0 0 0

Nonomuraea 0 0 0 0.0360 0.0010 0.0008 0.0011 0

Nordella 0.0374 0.0190 0.1214 0.0731 0.0010 0.0016 0.0701 0.0010

Novispirillum 0.0099 0.0060 0 0 0 0 0 0

Novosphingobium 0.8343 1.2179 0.8683 1.6486 0.3976 0.3754 2.0814 0.5854

Odoribacter 0 0 0 0 0.0325 0 0 0

Ochrobactrum 0.0033 0.0050 0 0 0 0 0 0

Oenothera 0.1431 0.0771 0 0.0535 0.0946 0.0294 0.1458 0.0226

Ohtaekwangia 0.0209 0.0481 0.1515 0.0830 0.0061 0.0016 0.0011 0

Olsenella 0 0 0 0 0 0 0 0.0334

Opitutus 0.0022 0.0180 0 0 0 0 0 0

Owenweeksia 0.0165 0.0080 0.1549 0.0972 0 0 0.0011 0

Oxalobacter 0.0044 0.0030 0 0 0 0 0 0

Paenibacillus 0.0671 0.0921 0.0012 0.0622 0 0.0857 0.0294 0.0010

Paludibacter 0.0528 0.0791 0 0 0 0 0 0

Paludibacterium 0.0066 0.0060 0 0 0 0 0 0

Pantoea 0.2950 0.1973 0 0 0 0 0 0

Parachlamydia 0.0044 0.0030 0.0994 0 0 0.0008 0 0

Paracoccus 0.0187 0.0080 0.0012 0.0022 0.5125 0.0016 0.0723 0.0216

Paracraurococcus 0.0044 0.0010 0 0 0 0 0 0

Treatments (mg Cl-/L) Genera 3 50 300 2500 Parasegetibacter 0.0044 0.0180 0.1630 0 0.0020 0.0437 0.0463 0.0030

Parvibaculum 0 0 0 0 0 0 0.0102 0.0010

Parvularcula 0 0.0030 0 0 0 0 0 0

Paucibacter 0.2421 0.8082 0 0 0 0 0 0

Pasteuria 0 0 0.0798 0.0502 0 0 0 0

Paucibacter 0 0 0.4544 0.4989 0.2919 0.3286 0.0260 0.0551

Paucimonas 0 0 0 0 0 0.0310 0 0.0148

Pectobacterium 0.1277 0.1492 0 0 0.0285 0.0302 0 0

Pedobacter 0.0473 0.3996 0.3561 0.0961 0.1403 0.0008 0.7074 0.0030

Pedomicrobium 0.0033 0.0010 0 0 0 0 0 0

Pedosphaera 0 0 0.0671 0.1015 0.0010 0.0008 0.0011 0

Pelobacter 0.0033 0 0 0 0 0 0 0

Pelomonas 0.1079 0.2454 0.2590 0.1256 0.0437 0.0508 0.0667 0.0315

Pelosinus 0.0088 0.0070 0.1468 0.0011 0 0 0 0.0010

Pelotomaculum 0.0198 0.0130 0.0104 0.3614 0 0.1151 0.0023 0

Peptoniphilus 0 0 0 0 0.0427 0 0 0

Peptostreptococcus 0 0 0 0 0.0183 0 0 0

Peredibacter 0.0176 0 0 0 0.1505 0 0 0

Phenylobacterium 0.1189 0.0971 0.0023 0.0469 0.0946 0.0873 0.0011 0.0502

Phormidium 0.0253 0.0010 0 0 0 0 0 0

Phycisphaera 0.0748 0.0371 0.1965 0.1114 0.3356 0.0643 0 0.0020

Phyllobacterium 0.0022 0.0030 0 0 0 0 0 0

Phytophthora 0.0044 0.0060 0.0173 0.0076 0.0153 0.0040 0.0011 0

Picochlorum 0 0.0050 0 0 0 0 0 0

Pirellula 0.1662 0.0911 0.4752 0.5033 0.1769 0.1587 0.0938 0.1712

Planctomyces 0.0693 0.0290 0.1411 0.1834 0.2563 0.0802 0.0554 0.0167

Planococcus 0 0 0 0 0 0 0 0.0010

Planomicrobium 0.0044 0.0030 0 0 0 0 0 0

Pleomorphobacterium 0 0 0 0 0.0010 0.0008 0 0

Pleomorphomonas 0.0044 0.0050 0 0 0 0 0 0

Polaromonas 0.1453 0.0971 0.0058 0.0011 0.1525 0.1278 0.1119 0.0177

Polyangium 0 0 0 0 0 0.0405 0 0

Porphyrobacter 0.0594 0.0270 0.0301 0.1234 0.0447 0.0008 0.0418 0.0944

Prevotella 0 0 0.0012 0.0852 0.0620 0 0 0.0148

Prochlorococcus 0.0077 0.0040 0 0.0229 0 0 0 0

Propionibacterium 0.0165 0.0070 0.0497 0.1856 0.4180 0.0381 0.0441 0.0964

Propionicimonas 0 0.0030 0 0 0 0.0183 0 0

Treatments (mg Cl-/L) Genera 3 50 300 2500 Propionigenium 0.0044 0.0010 0 0 0 0 0 0

Prosthecobacter 0.0088 0.0260 0.2046 0.4454 0.0010 0.0008 0.0011 0

Proteus 0 0 0 0 0 0.0032 0.0011 0

Prototheca 0 0.0030 0 0 0 0 0 0

Pseudoduganella 0 0 0.0035 0.1354 0.0010 0 0 0.0010

Pseudolabrys 0.0055 0 0 0 0 0 0 0

Pseudomonas 2.9837 4.4107 1.9042 1.5623 1.3983 0.6992 6.5708 0.7044

Pseudonocardia 0 0.0080 0 0.0011 0.2441 0.0317 0.0023 0.0020

Pseudospirillum 0 0 0.0012 0 0 0.0008 0 0.0236

Pseudoxanthomonas 0.0352 0.0060 0 0 0.0010 0 0 0

Quadrisphaera 0.0198 0.0451 0.0127 0 0 0 0 0

Quatrionicoccus 0.0011 0.0050 0 0 0 0 0 0

Rahnella 0.1244 0.3215 0 0 0 0 0 0

Ralstonia 0.0066 0.0090 0.0046 0.0022 0.8481 0.0024 0.0011 0.0020

Ramlibacter 0 0 0.0474 0.0011 0.0010 0.0095 0 0

Reyranella 0.0077 0.0030 0 0.0011 0.0620 0 0 0

Rheinheimera 0.0033 0.0210 0 0 0 0 0 0

Rhizobacter 1.1600 1.3290 0 0 0 0 0 0

Rhizobium 11.029 9.2080 6.2120 6.2177 9.1860 1.7231 3.0634 0.8146

Rhizomicrobium 0.0077 0.0010 0 0 0 0 0 0

Rhodanobacter 0 0 0.0012 0.0011 0.0031 0 0 0

Rhodobacter 1.7610 1.1638 2.7944 2.9380 0.9132 1.2754 2.0600 0.3866

Rhodobium 0.0099 0.0020 0.0671 0 0.0010 0 0.0395 0.0010

Rhodocista 0.0528 0.0601 0 0 0.1302 0 0 0

Rhodococcus 0.0704 0.0341 0.0012 0.0011 0.1993 0 0.0746 0

Rhodocyclus 0.0451 0.0601 0 0 0 0 0 0

Rhodoferax 1.6498 1.5614 0.4856 0.9750 0.4027 0.0698 0.0689 0.0610

Rhodomicrobium 0.2102 0.1843 0.1237 0.0579 0.0020 0.0024 0.0463 0

Rhodopirellula 0 0.0030 0 0 0 0 0 0.0266

Rhodoplanes 0.0044 0.0100 0.0890 0.0448 0 0 0.2102 0.0010

Rhodopseudomonas 0.0055 0.0040 0 0 0 0 0 0

Rhodospirillum 0.0220 0.0671 0.1353 0 0 0 0.0011 0.0010

Rhodovastum 0.0077 0 0 0 0 0 0 0

Rhodovulum 0.0044 0.0070 0.0012 0 0 0 0 0

Rickettsia 0.0275 0.0611 0.0069 0.0590 0 0 0 0

Rickettsiella 0 0 0 0.0502 0 0 0 0.0010

Roseateles 0.1882 0.2304 0.0023 0.2708 0.0112 0.0016 0.0565 0.0246

Treatments (mg Cl-/L) Genera 3 50 300 2500 Roseibacillus 0.0077 0.0190 0.0925 0.0622 0.0854 0.0833 0.0655 0.0108

Roseibacterium 0 0 0.0671 0.0011 0 0 0 0

Roseiflexus 0 0.0100 0 0 0 0 0.0056 0

Roseimaritima 0 0.0030 0 0 0 0 0 0

Roseobacter 0.0033 0 0 0 0 0 0 0

Roseococcus 0.0055 0.0010 0.0902 0 0 0 0 0

Roseomonas 0.0561 0.0551 0.0578 0.1125 0.0529 0.0333 0.0701 0.0010

Rubellimicrobium 0 0 0 0 0.1342 0 0 0

Rubrivivax 0.1706 0.2263 0.2359 0.2347 0.2766 0.1222 0.0203 0.0561

Rubrobacter 0 0 0.0416 0 0.1993 0.0008 0 0.0010

Runella 0.0396 0.0621 0.3087 0.1321 0.2868 0.0016 0.0023 0.0010

Saccharibacillus 0.0088 0.0020 0 0 0 0 0 0

Saccharibacter 0 0.0060 0 0 0 0 0.0362 0

Saccharospirillum 0.0044 0.0090 0 0 0 0 0 0

Saccharopolyspora 0 0 0 0.0011 0 0.0016 0 0

Sandaracinus 0.0055 0.0030 0.0058 0 0 0 0 0

Sandarakinorhabdus 0 0.0080 0 0.0011 0.0834 0 0 0

Sanguibacter 0.1563 0.1272 0 0 0 0 0 0

Schlegelella 0 0 0 0 0 0 0.0486 0

Sediminibacterium 0.0693 0.2083 0.1214 0.1179 0.0020 0.0016 0.0011 0.0010

Serratia 0 0 0 0 0 0.0008 0 0.0256

Shigella 0 0 0.0012 0.0022 0.2430 0.0373 0.4667 0.1407

Sideroxydans 0 0.0040 0 0 0 0 0 0

Simaetha 0 0.0030 0 0 0 0 0 0

Simplicispira 0.0099 0.0290 0.0197 0.0044 0.0061 0 0.0237 0.0010

Singulisphaera 0 0 0 0 0.0447 0 0 0

Sinorhizobium 0.0033 0.0110 0 0 0 0 0 0

Smithella 0 0.0030 0 0 0 0 0 0

Solibacillus 0 0.0050 0 0 0 0 0 0

Solirubrobacter 0 0 0 0 0 0 0.0056 0

Sorangium 0.0110 0 0 0 0 0 0.0542 0

Sphaerobacter 0 0 0.1260 0 0.0478 0.0008 0 0.0010

Sphaerotilus 0.4017 0.2904 0.4312 0.6223 0.4607 0.1000 0.0045 0.0698

Sphingobacterium 0.0451 0.0631 0.3584 0.2948 0.0600 0.0016 0.0023 0.0030

Sphingobium 0.3918 0.7241 0.1676 0.4072 0.0081 0.1254 1.2927 0.4102

Sphingomonas 0.4986 0.5128 0.5469 0.1146 0.8278 0.0683 0.1492 0.1633

Sphingopyxis 0.8331 0.5919 1.1851 1.4717 0.9529 0.5286 0.7458 0.3915

Treatments (mg Cl-/L) Genera 3 50 300 2500 Sphingorhabdus 0.0264 0.0200 0.1804 0.0371 0.0885 0.0746 0.0497 0.0846

Spirosoma 0.0121 0 0 0 0.1515 0.0143 0 0

Sporichthya 0.0143 0.0040 0 0 0 0.0008 0 0.0403

Sporobacter 0 0.0060 0 0 0 0 0 0

Staphylococcus 0.0143 0.0100 0.1214 0.1146 1.0586 0.0040 0.0034 0.0089

Starkeya 0.0154 0.0030 0.0023 0.0033 0.0031 0 0 0

Stenotrophomonas 0.5250 0.2754 0.2798 0.1943 0.0285 0.1683 0.0034 0.0020

Steroidobacter 0.0209 0.0300 0.0775 0.0033 0 0.0349 0.0011 0

Sterolibacterium 0.2179 0.1813 0.2509 0.0426 0.0010 0.0683 0 0

Streptococcus 0 0 0.0012 0.6780 1.4532 0.0008 0.1017 0.0885

Streptomyces 0 0 0.0012 0.1026 0.1271 0.0024 0.0011 0.0020

Sulfitobacter 0 0 0 0 0.0010 0 0 0.0020

Sulfurimonas 0 0 0 0.0011 0.0681 0.0008 0.0011 0.0010

Sulfurovum 0 0 0.0197 0.0011 0.1078 0.1198 0 0.0010

Sulfurospirillum 0.0044 0.0010 0 0 0 0 0 0

Synechococcus 0 0 0 0 0 0 0 0.0039

Syntrophus 0 0 0.1538 0 0 0 0 0.0010

Tabrizicola 0.3973 0.5398 0 0 0 0 0 0

Tepidamorphus 0 0 0 0 0.0549 0 0 0.0010

Tepidimicrobium 0 0 0 0 0 0 0 0.0010

Tepidimonas 0 0 0 0.0797 0.0031 0 0.0011 0.0010

Terrabacter 0 0.0040 0 0 0 0 0 0

Terrimonas 0.0429 0.0851 0.7700 0.7151 0.4963 0.0024 0.0203 0.0020

Tessaracoccus 0 0 0 0 0.0020 0.0008 0.0011 0

Tetrasphaera 0 0 0 0 0.0590 0 0.0011 0.0216

Thalassospira 0.0121 0.0040 0 0 0 0 0 0

Thauera 0.0033 0.0030 0 0 0 0 0 0

Thermanaerothrix 0.0231 0.0190 0 0 0 0 0 0

Thermicanus 0 0.0030 0.0012 0 0 0 0 0

Thermoflavimicrobium 0 0 0.0936 0 0 0.0008 0 0

Thermomonas 0.1079 0.2704 0.0994 0.1485 0.0742 0.1151 0.0011 0.0020

Thermovum 0.0022 0.0010 0 0 0 0 0 0

Thermus 0 0.0030 0.0382 0 0 0 0 0

Thioalkalivibrio 0 0.0030 0 0 0.0010 0.0246 0 0

Thiohalophilus 0 0 0 0 0.0702 0.0008 0 0.0010

Thiomicrospira 0 0 0 0 0 0.0056 0 0

Thioprofundum 0 0.0030 0 0 0 0.0008 0 0.0039

Treatments (mg Cl-/L) Genera 3 50 300 2500 Thiorhodospira 0.0044 0.0020 0 0 0 0 0 0

Thiothrix 0 0 0 0.2020 0.0010 0.0016 0 0.0010

Tissierella 0 0 0.0567 0 0 0 0 0.0010

Tolumonas 1.3042 1.3991 0.1087 0.0721 0 0.0008 0 0.0010

Trichococcus 0.0682 0.2474 0.2370 0.9215 0.1261 0.0754 0.2407 0.0620

Turicibacter 0 0 0 0.0022 0 0 0 0.0010

Uliginosibacterium 3.7464 2.3526 0.2983 0.1583 0.0254 0.1127 0.0203 0.0069

Undibacterium 0.0231 0.022 0.0786 0 0.0407 0.0579 0.0011 0.0010

Variovorax 0.0792 0.1002 0.5422 0.3395 0.4393 0.1040 0.3944 0.0472

Veillonella 0 0 0 0.0917 0 0 0.0011 0.0394

Verrucomicrobium 0.0341 0.0541 0.6347 0.4269 0.0031 0.2357 0 0

Vibrio 0.0121 0 0 0 0 0 0 0

Williamsia 0 0 0 0 0.0264405 0 0 0

Wolbachia 0 0 0.2312 0.0317 0 0.0008 0 0

Woodsholea 0 0 0 0.0011 0.0519 0.0008 0 0

Xanthomonas 0.0011 0.0240 0.1977 0 0 0.0119 0 0.0010

Xylella 0.0121 0.0050 0 0 0 0 0 0

Yonghaparkia 0.2190 0.0981 0 0 0 0 0 0

Zavarzinella 0.0011 0.0070 0 0 0 0 0 0.0384

Zea 0 0 0 0.0469 0 0 0 0

Zebrasoma 0 0.0050 0 0 0 0 0 0

Zoogloea 0.0187 0.0431 0 0 0 0 0 0

APPENDIX D

Bacteria phyla relative abundance (percentage) data for all treatments (Two replicates are indicated per treatment)

Treatments (mg Cl-/L) Phyla 3 3 50 50 300 300 2500 2500 Acetothermia 0 0 0 0.0011 0 0 0 0 Acidobacteria 0.0451 0.0250 0.6440 0.2162 0.2034 0.1333 0 0.0020 Actinobacteria 6.4110 3.5664 6.0074 5.1401 12.3365 4.2851 6.1482 3.0615 Apicomplexa 0 0 0 0 0 0 0.0034 0 Aquificae 0 0 0.0012 0 0.1647 0 0 0 Armatimonadetes 0 0 0 0 0 0 0.0599 0 Armatimonadetes 0 0.0180 0 0 0 0 0 0 Arthropoda 0 0.0030 0 0 0 0 0 0 Ascomycota 0.0044 0.0040 0 0 0 0 0 0 Bacteroidetes 34.6346 36.8841 31.3421 34.1267 22.7165 21.4515 28.4688 11.4434 Candidatus saccharibacteria 0 0.0080 0.0254 0.1004 0.2705 0.0325 0.0362 0.0010 Chlamydiae 0.0803 0.0611 0.2786 0 0.1220 0.2556 0 0.0541 Chloroflexi 0.0913 0.2354 0.3781 0.1245 0.3610 0.0421 0.3333 0.0256 Chlorophyta 0.0066 0.0080 0 0 0 0 0 0 Chordata 0 0.0050 0 0 0 0 0 0 Cyanobacteria 2.0867 1.2809 0.3573 0.0284 0.4241 0.4024 2.6272 0.2843 Deinococcus_thermus 0 0.0030 0.0439 0.0044 0.0631 0.3691 0.0723 1.2661 Eukaryota 0.0209 0.0280 0.0173 0.0076 0.0153 0.0040 0.0011 0 Euryarchaeota 0 0 0.0462 0 0.0580 0 0 0 Fibrobacteres 0.0066 0 0 0 0 0 0 0 Firmicutes 0.4744 0.6550 1.5724 3.0057 22.5609 5.5661 24.3929 10.3898 Fusobacteria 0.0088 0.0010 0 0 0 0 0 0 Gemmatimonadetes 0.0528 0.0140 0 0.1103 0.1220 0 0 0.0010 Ignavibacteriae 0 0 0 0.1256 0 0 0 0 Nitrospinae 0 0 0 0 0 0 0 0.0010 Nitrospirae 0.0011 0.0080 0 0 0 0 0 0 Planctomycetes 0.3918 0.2083 1.3562 0.9629 1.0637 0.3770 0.1503 0.2617 Proteobacteria 55.2025 56.3236 50.7006 46.9922 37.9492 65.0965 33.1107 72.9950 Spirochaetes 0 0 0 0 0 0 0 0 Streptophyta 0.1662 0.0771 0 0.1004 0.0946 0.0310 0.1492 0.0236 Tenericutes 0.0671 0.0120 0.0035 0.2773 0.8064 0.0944 0.4520 0.0571 Thaumarchaeota 0 0 0.0023 0.0022 0.0031 0.0024 0.0407 0 Verrucomicrobia 0.2476 0.5709 7.2236 8.6741 0.6651 1.8572 3.9538 0.1328

APPENDIX E

Supplemental Figures

First Supplemental Figure. Venn Diagram of bacteria genera in all treatments from Experiment II.

5 3 mg Cl-/L 4 50 mg Cl-/L Mortalities 3 300 mg Cl-/L 1000 mg Cl-/L 2 2500 mg Cl-/L 1

Dates

Second Supplemental Figure. Time series of amphipod mortalities in Experiment III.

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