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Shifts in microbial community structure and function in surface waters impacted by unconventional oil and gas wastewater revealed by metagenomics

Unconventional oil and gas (UOG) production produces large quantities of wastewater with complex geochemistry and largely uncharacterized impacts on surface waters. In this study, we assessed shifts in microbial community structure and function in sediments and waters upstream and downstream from a UOG wastewater disposal facility. To do this, quantitative PCR for 16S rRNA and antibiotic resistance genes along with metagenomic sequencing were performed. Elevated conductivity and markers of UOG wastewater characterized sites sampled downstream from the disposal facility compared to background sites. Shifts in overall high level functions and microbial community structure were observed between background sites and downstream sediments. Increases in Deltaproteobacteria and Methanomicrobia and decreases in Thaumarchaeota were observed at downstream sites. Genes related to dormancy and sporulation and methanogenic respiration were 18–86 times higher at downstream, impacted sites. The potential for these sediments to serve as reservoirs of antimicrobial resistance was investigated given frequent reports of the use of biocides to control the growth of nuisance bacteria in UOG operations. A shift in resistance profiles downstream of the UOG facility was observed including increases in acr B and mex B genes encoding for multidrug efflux pumps, but not overall abundance of resistance genes. The observed shifts in microbial community structure and potential function indicate changes in respiration, nutrient cycling, and markers of stress in a stream impacted by UOG waste disposal operations.

West Virginia

Genomically diverse carbapenem resistant Enterobacteriaceae from wild birds provide insight into global patterns of spatiotemporal dissemination

Carbapenem resistant Enterobacteriaceae (CRE) are a threat to public health globally, yet the role of the environment in the epidemiology of CRE remains elusive. Given that wild birds can acquire CRE, likely from foraging in anthropogenically impacted areas, and may aid in the maintenance and dissemination of CRE in the environment, a spatiotemporal comparison of isolates from different regions and timepoints may be useful for elucidating epidemiological information. Thus, we characterized the genomic diversity of CRE from fecal samples opportunistically collected from gulls ( Larus spp.) inhabiting Alaska (USA), Chile, Spain, Turkey, and Ukraine and from black kites ( Milvus migrans ) sampled in Pakistan and assessed evidence for spatiotemporal patterns of dissemination. Within and among sampling locations, a high diversity of carbapenemases was found, including Klebsiella pneumoniae carbapenemase (KPC), New Delhi metallo-beta-lactamase (NDM), oxacillinase (OXA), and Verona integron Metallo beta-lactamase (VIM). Although the majority of genomic comparisons among samples did not provide evidence for spatial dissemination, we did find strong evidence for dissemination among Alaska, Spain, and Turkey. We also found strong evidence for temporal dissemination among samples collected in Alaska and Pakistan, though the majority of CRE clones were transitory and were not repeatedly detected among locations where samples were collected longitudinally. Carbapenemase-producing hypervirulent K. pneumoniae was isolated from gulls in Spain and Ukraine and some isolates harbored antimicrobial resistance genes conferring resistance to up to 10 different antibiotic classes, including colistin. Our results are consistent with local acquisition of CRE by wild birds with spatial dissemination influenced by intermediary transmission routes, likely involving humans. Furthermore, our results support the premise that anthropogenically-associated wild birds may be good sentinels for understanding the burden of clinically-relevant antimicrobial resistance in the local human population.

Science of the Total Environment

Factors related to occurrence and distribution of selected bacterial and protozoan pathogens in Pennsylvania streams

The occurrence and distribution of fecal indicator bacteria (FIB) and bacterial and protozoan pathogens are controlled by diverse factors. To investigate these factors in Pennsylvania streams, 217 samples were collected quarterly from a 27-station water-quality monitoring network from July 2007 through August 2009. Samples were analyzed for concentrations of Escherichia coli (EC) and enterococci (ENT) indicator bacteria, concentrations of Cryptosporidium oocysts and Giardia cysts, and the presence of four genes related to pathogenic types of EC ( eae A, stx 2, stx 1, rfb O157 ) plus three microbial source tracking (MST) gene markers that are also associated with pathogenic ENT and EC ( esp , LTIIa, STII). Water samples were concurrently analyzed for basic water chemistry, physical measures of water quality, nutrients, metals, and a suite of 79 organic compounds that included hormones, pharmaceuticals, and antibiotics. For each sample location, stream discharge was measured by using standardized methods at the time of sample collection, and ancillary sample site information, such as land use and geological characteristics, was compiled. Samples exceeding recreational water quality criteria were more likely to contain all measured pathogen genes but not Cryptosporidium or Giardia (oo)cysts. FIB and Giardia density and frequency of eae A gene occurrence were significantly related to season. When discharge at a sampling location was high (>75th percentile of daily mean discharge), there were greater densities of FIB and Giardia , and the stx2 , rfb O157 , STII, and esp genes were found more frequently than at other discharge conditions. Giardia occurrence was likely related to nonpoint sources, which are highly influential during seasonal overland transport resulting from snowmelt and elevated precipitation in late winter and spring in Pennsylvania. When MST markers of human, swine, or bovine origin were present, samples more frequently carried the eae A, stx 2, stx 1, and rfb O157 genes, but no genes were related exclusively to an individual MST marker. The human source pharmaceuticals (HSPs) acetaminophen and caffeine were correlated with Giardia , and the presence of HSPs proved to be more useful than MST markers in distinguishing the occurrence of Giardia . The HSPs caffeine and carbamazepine were correlated with the sum total of pathogen genes detected in a sample, demonstrating the value of using HSPs as an indicator of fecally derived pathogens. Sites influenced by urban land use with less forest were more likely to have greater FIB and Giardia densities and sum of the array of pathogen genes. Sites dominated by shallow carbonate bedrock in the upstream catchment were likely to have greater FIB densities and higher sum totals of pathogen genes but no correlation with Giardia detection. Our study provides a range of specific environmental, chemical, geologic, and land-use variables related to occurrence and distribution of FIB and selected bacterial and protozoan pathogens in Pennsylvania streams. The information presented could be useful for resource managers in understanding bacterial and protozoan pathogen occurrence and their relation to fecal indicator bacteria in similar settings.

Pennsylvania

Food, beverage, and feedstock processing facility wastewater: A unique and underappreciated source of contaminants to U.S. streams

Process wastewaters from food, beverage, and feedstock facilities, although regulated, are an under-investigated environmental contaminant source. Food process wastewaters (FPWWs) from 23 facilities in 17 U.S. states were sampled and documented for a plethora of chemical and microbial contaminants. Of the 576 analyzed organics, 184 (32%) were detected at least once, with concentrations as large as 143 μg L –1 (6:2 fluorotelomer sulfonic acid), and as many as 47 were detected in a single FPWW sample. Cumulative per/polyfluoroalkyl substance concentrations up to 185 μg L –1 and large pesticide transformation product concentrations (e.g., methomyl oxime, 40 μg L –1 ; clothianidin TMG, 2.02 μg L –1 ) were observed. Despite 48% of FPWW undergoing disinfection treatment prior to discharge, bacteria resistant to third-generation antibiotics were found in each facility type, and multiple bacterial groups were detected in all samples, including total coliforms. The exposure–activity ratios and toxicity quotients exceeded 1.0 in 13 and 22% of samples, respectively, indicating potential biological effects and toxicity to vertebrates and invertebrates associated with the discharge of FPWW. Organic contaminant profiles of FPWW differed from previously reported contaminant profiles of municipal effluents and urban storm water, indicating that FPWW is another important source of chemical and microbial contaminant mixtures discharged into receiving surface waters.

Alabama, Arkansas, California, Idaho, Illinois, Io

Contaminant exposure and transport from three potential reuse waters within a single watershed

Global demand for safe and sustainable water supplies necessitates a better understanding of contaminant exposures in potential reuse waters. In this study, we compared exposures and load contributions to surface water from the discharge of three reuse waters (wastewater effluent, urban stormwater, and agricultural runoff). Results document substantial and varying organic-chemical contribution to surface water from effluent discharges (e.g., disinfection byproducts [DBP], prescription pharmaceuticals, industrial/household chemicals), urban stormwater (e.g., polycyclic aromatic hydrocarbons, pesticides, nonprescription pharmaceuticals), and agricultural runoff (e.g., pesticides). Excluding DBPs, episodic storm-event organic concentrations and loads from urban stormwater were comparable to and often exceeded those of daily wastewater-effluent discharges. We also assessed if wastewater-effluent irrigation to corn resulted in measurable effects on organic-chemical concentrations in rain-induced agricultural runoff and harvested feedstock. Overall, the target-organic load of 491 g from wastewater-effluent irrigation to the study corn field during the 2019 growing season did not produce substantial dissolved organic-contaminant contributions in subsequent rain-induced runoff events. Out of the 140 detected organics in source wastewater-effluent irrigation, only imidacloprid and estrone had concentrations that resulted in observable differences between rain-induced agricultural runoff from the effluent-irrigated and nonirrigated corn fields. Analyses of pharmaceuticals and per-/polyfluoroalkyl substances in at-harvest corn-plant samples detected two prescription antibiotics, norfloxacin and ciprofloxacin, at concentrations of 36 and 70 ng/g, respectively, in effluent-irrigated corn-plant samples; no contaminants were detected in noneffluent irrigated corn-plant samples.

Oklahoma

Escherichia coli and enterococci at beaches in the Grand Traverse Bay, Lake Michigan: Sources, characteristics, and environmental pathways

This study quantified Escherichia coli(EC) and enterococci (ENT) in beach waters and dominant source materials, correlated these with ambient conditions, and determined selected EC genotypes and ENT phenotypes. Bathing-water ENT criteria were exceeded more frequently than EC criteria, providing conflicting interpretations of water quality. Dominant sources of EC and ENT were bird feces (108/d/bird), storm drains (107/d), and river water (1011/d); beach sands, shallow groundwater and detritus were additional sources. Beach-water EC genotypes and ENT phenotypes formed clusters with those from all source types, reflecting diffuse inputs. Some ENT isolates had phenotypes similar to those of human pathogens and/or exhibited high-level resistance to human-use antibiotics. EC and ENT concentrations were influenced by collection time and wind direction. There was a 48-72-h lag between rainfall and elevated EC concentrations at three southern shoreline beaches, but no such lag at western and eastern shoreline beaches, reflecting the influence of beach orientation with respect to cyclic (3-5 d) summer weather patterns. In addition to local contamination sources and processes, conceptual or predictive models of Great Lakes beach water quality should consider regional weather patterns, lake hydrodynamics, and the influence of monitoring method variables (time of day, frequency).

Grand Traverse Bay

Comparison of seven protocols to identify fecal contamination sources using Escherichia coli

Microbial source tracking (MST) uses various approaches to classify fecal-indicator microorganisms to source hosts. Reproducibility, accuracy, and robustness of seven phenotypic and genotypic MST protocols were evaluated by use of Escherichia coli from an eight-host library of known-source isolates and a separate, blinded challenge library. In reproducibility tests, measuring each protocol's ability to reclassify blinded replicates, only one (pulsed-field gel electrophoresis; PFGE) correctly classified all test replicates to host species; three protocols classified 48-62% correctly, and the remaining three classified fewer than 25% correctly. In accuracy tests, measuring each protocol's ability to correctly classify new isolates, ribotyping with EcoRI and PvuII approached 100% correct classification but only 6% of isolates were classified; four of the other six protocols (antibiotic resistance analysis, PFGE, and two repetitive-element PCR protocols) achieved better than random accuracy rates when 30-100% of challenge isolates were classified. In robustness tests, measuring each protocol's ability to recognize isolates from nonlibrary hosts, three protocols correctly classified 33-100% of isolates as "unknown origin," whereas four protocols classified all isolates to a source category. A relevance test, summarizing interpretations for a hypothetical water sample containing 30 challenge isolates, indicated that false-positive classifications would hinder interpretations for most protocols. Study results indicate that more representation in known-source libraries and better classification accuracy would be needed before field application. Thorough reliability assessment of classification results is crucial before and during application of MST protocols.

Environmental Science & Technology

What makes a natural clay antibacterial?

Natural clays have been used in ancient and modern medicine, but the mechanism(s) that make certain clays lethal against bacterial pathogens has not been identified. We have compared the depositional environments, mineralogies, and chemistries of clays that exhibit antibacterial effects on a broad spectrum of human pathogens including antibiotic resistant strains. Natural antibacterial clays contain nanoscale (<200 nm), illite-smectite and reduced iron phases. The role of clay minerals in the bactericidal process is to buffer the aqueous pH and oxidation state to conditions that promote Fe 2+ solubility. Chemical analyses of E. coli killed by aqueous leachates of an antibacterial clay show that intracellular concentrations of Fe and P are elevated relative to controls. Phosphorus uptake by the cells supports a regulatory role of polyphosphate or phospholipids in controlling Fe 2+ . Fenton reaction products can degrade critical cell components, but we deduce that extracellular processes do not cause cell death. Rather, Fe 2+ overwhelms outer membrane regulatory proteins and is oxidized when it enters the cell, precipitating Fe 3+ and producing lethal hydroxyl radicals.

Environmental Science & Technology

Fate of sulfamethoxazole, 4-nonylphenol, and 17β-estradiol in groundwater contaminated by wastewater treatment plant effluent

Organic wastewater contaminants (OWCs) were measured in samples collected from monitoring wells located along a 4.5-km transect of a plume of groundwater contaminated by 60 years of continuous rapid infiltration disposal of wastewater treatment plant effluent. Fifteen percent of the 212 OWCs analyzed were detected, including the antibiotic sulfamethoxazole (SX), the nonionic surfactant degradation product 4-nonylphenol (NP), the solvent tetrachloroethene (PCE), and the disinfectant 1,4-dichlorobenzene (DCB). Comparison of the 2005 sampling results to data collected from the same wells in 1985 indicates that PCE and DCB are transported more rapidly in the aquifer than NP, consistent with predictions based on compound hydrophobicity. Natural gradient in situ tracer experiments were conducted to evaluate the subsurface behavior of SX, NP, and the female sex hormone 17&beta;-estradiol (E2) in two oxic zones in the aquifer: (1) a downgradient transition zone at the interface between the contamination plume and the overlying uncontaminated groundwater and (2) a contaminated zone located beneath the infiltration beds, which have not been loaded for 10 years. In both zones, breakthrough curves for the conservative tracer bromide (Br &minus; ) and SX were nearly coincident, whereas NP and E2 were retarded relative to Br &minus; and showed mass loss. Retardation was greater in the contaminated zone than in the transition zone. Attenuation of NP and E2 in the aquifer was attributed to biotransformation, and oxic laboratory microcosm experiments using sediments from the transition and contaminated zones show that uniform-ring-labeled 14 C 4-normal-NP was biodegraded more rapidly (30&minus;60% recovered as 14 CO 2 in 13 days) than 4- 14 C E2 (20&minus;90% recovered as 14 CO 2 in 54 days). There was little difference in mineralization potential between sites.

Massachusetts

Fungal impacts on Earth’s ecosystems

Over the past billion years, the fungal kingdom has diversified to more than two million species, with over 95% still undescribed. Beyond the well-known macroscopic mushrooms and microscopic yeast, fungi are heterotrophs that feed on almost any organic carbon, recycling nutrients through the decay of dead plants and animals and sequestering carbon into Earth’s ecosystems. Human-directed applications of fungi extend from leavened bread, alcoholic beverages and biofuels to pharmaceuticals, including antibiotics and psychoactive compounds. Conversely, fungal infections pose risks to ecosystems ranging from crops to wildlife to humans; these risks are driven, in part, by human and animal movement, and might be accelerating with climate change. Genomic surveys are expanding our knowledge of the true biodiversity of the fungal kingdom, and genome-editing tools make it possible to imagine harnessing these organisms to fuel the bioeconomy. Here, we examine the fungal threats facing civilization and investigate opportunities to use fungi to combat these threats.

Nature

Gulls as sources of environmental contamination by colistin-resistant bacteria

In 2015, the mcr-1 gene was discovered in Escherichia coli in domestic swine in China that conferred resistance to colistin, an antibiotic of last resort used in treating multi-drug resistant bacterial infections in humans. Since then, mcr-1 was found in other human and animal populations, including wild gulls. Because gulls could disseminate the mcr-1 gene, we conducted an experiment to assess whether gulls are readily colonized with mcr-1 positive E. coli, their shedding patterns, transmission among conspecifics, and environmental deposition. Shedding of mcr-1 E. coli by small gull flocks followed a lognormal curve and gulls shed one strain >101 log10 CFU/g in their feces for 16.4 days, which persisted in the environment for 29.3 days. Because gulls are mobile and can shed antimicrobial-resistant bacteria for extended periods, gulls may facilitate transmission of mcr-1 positive E. coli to humans and livestock through fecal contamination of water, public areas and agricultural operations.

Scientific Reports

Validation and application of a standardized quantitative PCR assay for the assessment of antimicrobial resistance genes in surface water

Antimicrobial resistance can be an indicator of anthropogenic contamination in surface waters and is a potential public health threat. Methodological standardization for characterization of antimicrobial resistance in the environment is lacking. Quantitative PCR (qPCR) is used for rapid assessment of antibiotic resistance genes (ARGs) from environmental sources, including surface water. Here we describe the validation and application of a qPCR assay for 47 bacterial gene targets intended for surface water samples. The qPCR assay displayed excellent sensitivity (97.66%) and specificity (98.71%) for detecting ARGs when compared to whole genome sequencing of bacterial isolates. The qPCR assay was able to detect up to 6/8 (75.0%) of ARGs spiked into sterile water at varying concentrations and four sample ultrafiltration volumes. Nineteen different ARGs were detected across six samples sites at three national parks in Alaska using ultrafiltered surface water samples. The number of unique ARGs detected was higher at sites within parks with greater visitation. The relative abundance of ARGs/16S from Exit Creek in Kenai Fjords National Park, downstream from a visitor center was greater than all other sampled sites. We have demonstrated a robust qPCR assay for monitoring ARGs in surface waters, including those that are minimally human impacted.

Alaska

Assessing microplastics, per- and polyfluoroalkyl substances (PFAS), and other contaminants of global concern in wadable agricultural streams

Microplastics, per- and polyfluoroalkyl substances (PFAS), antibiotic resistance genes (ARGs), pharmaceuticals and personal care products (PPCPs), and pesticides may lead to unintended environmental contamination through many pathways in multiple matrices. This statewide, multi-matrix study of contaminants of global concern (CGCs) in agricultural streams across Iowa (United States) is the first to examine multiple CGCs in water, bed sediment, and fish to understand their occurrence in small streams located in regions of intense agriculture activity. Iowa plays a pivotal role in agriculture, with more than 85% of Iowa’s landscape devoted to agriculture making it an ideal location for determining the prevalence of CGCs to provide critical baseline exposure data. Fifteen sites were sampled across a range of predominant land uses (e.g., poultry, swine); all sites had detections of microplastics in all matrices. Concentrations of PFAS varied but were detected in water and sediment; all fish had detections of perfluorooctanesulfonate (PFOS), a type of PFAS. More than 50% of water and bed sediment samples had detections of ARGs. The most frequently detected PPCP was metformin. No sites had a cumulative exposure activity ratio greater than 1.0 for chemical exposures; 13 sites were above the 0.001 precautionary threshold. Toxicity quotients calculated using Aquatic Life Benchmarks were below the 0.1 moderate risk threshold for chemical exposures for all but one site. For fish, all sites exceeded the moderate and high-risk thresholds proposed for microplastic particles for food dilution (both chronic and acute exposures) and all sites exceeded the microplastic moderate threshold proposed for chronic tissue translocation, and two sites exceeded the threshold for acute tissue translocation.

Iowa

Thermal sensitivity of immune function: evidence against a generalist-specialist trade-off among endothermic and ectothermic vertebrates

Animal body temperature (Tbody) varies over daily and annual cycles, affecting multiple aspects of biological performance in both endothermic and ectothermic animals. Yet a comprehensive comparison of thermal performance among animals varying in Tbody (mean and variance) and heat production is lacking. Thus, we examined the thermal sensitivity of immune function (a crucial fitness determinant) in Vertebrata, a group encompassing species of varying thermal biology. Specifically, we investigated temperature-related variation in two innate immune performance metrics, hemagglutination and hemolysis, for 13 species across all seven major vertebrate clades. Agglutination and lysis were temperature dependent and were more strongly related to the thermal biology of species (e.g., mean Tbody) than to the phylogenetic relatedness of species, although these relationships were complex and frequently surprising (e.g., heterotherms did not exhibit broader thermal performance curves than homeotherms). Agglutination and lysis performance were positively correlated within species, except in taxa that produce squalamine, a steroidal antibiotic that does not lyse red blood cells. Interestingly, we found the antithesis of a generalist-specialist trade-off: species with broader temperature ranges of immune performance also had higher peak performance levels. In sum, we have uncovered thermal sensitivity of immune performance in both endotherms and ectotherms, highlighting the role that temperature and life history play in immune function across Vertebrata.

American Naturalist

Surgical implantation of transmitters into fish

Although the Animal Welfare Act does not cover poikilotherms, individual institutions and policies and legal requirements other than the Animal Welfare Act (e.g., the US Public Health Service and the Interagency Research Animal Committee's Principles for the Utilization and Care of Vertebrate Animals Used in Testing, Research, and Training) require the review of projects involving fish by institutional animal care and use committees (IACUCs). IACUCs may, however, lack the knowledge and experience to evaluate fish projects judiciously, especially when the projects are in field settings. Surgeries involving implantation of transmitters and other instruments into the coelom, which now comprise a very common research tool in the study of free-ranging fishes, are examples of surgeries that use a broad spectrum of surgical and anesthetic techniques, some of which would not be considered acceptable for similar work on mammals. IACUCs should apply the standards they would expect to be used for surgeries on homeotherms to surgeries on fish. Surgeons should be carefully trained and experienced. Surgical instruments and transmitters should be sterile. Regulations and laws on the use of drugs in animals should be followed, particularly those concerned with anesthetics and antibiotics used on free-ranging fish. Exceptions to surgical procedures should be made only when circumstances are extreme enough to warrant the use of less than optimal procedures.

ILAR Journal

Cnidarian–algal partnerships structure bacterial communities during strobilation in Cassiopea xamachana

Cnidarian–algal (Symbiodiniaceae) symbioses rely on complex interactions among the cnidarian host, algal symbionts, and associated bacterial communities. In the upside-down jellyfish Cassiopea xamachana , the polyp-to-medusa transition (strobilation) requires the establishment of symbiosis with Symbiodiniaceae algal partners, yet bacterial community dynamics during this developmental process remain unknown. Here, we experimentally induced symbiosis in aposymbiotic polyps using four algal treatments: xenic Symbiodinium microadriaticum (native symbiont), xenic Breviolum minutum , antibiotic-treated B. minutum , and a photosynthetically impaired B. minutum mutant. We combined 16S rRNA gene sequencing with measurements of photosynthetic efficiency, asexual budding, and algal surface N-glycan profiles to characterize holobiont assembly during symbiosis onset and strobilation. Algal treatment structured bacterial communities in both algal cultures and polyp tissues. Our analyses identified a set of amplicon sequence variants that consistently distinguished strobilating polyps from non-strobilating aposymbiotic and mutant polyps, in addition to potential bacterial biomarkers associated with successful metamorphosis. Strobilation was associated with the enrichment of bacterial communities putatively involved in sulfur and nitrogen cycling, whereas non-strobilating aposymbiotic and mutant polyps were characterized by opportunistic bacteria and increased community variability. Together, these results reveal coordinated changes in algal physiology, surface glycan profiles, and bacterial community structure associated with successful strobilation in C. xamachana and support a model in which tripartite host–alga–bacteria interactions influence cnidarian life stage transitions.

ISME Communications

Low occurrence of multi-antimicrobial and heavy metal resistance in Salmonella enterica from wild birds in the United States

Wild birds are common reservoirs of Salmonella enterica . Wild birds carrying resistant S . enterica may pose a risk to public health as they can spread the resistant bacteria across large spatial scales within a short time. Here, we whole-genome sequenced 375 S . enterica strains from wild birds collected in 41 U.S. states during 1978–2019 to examine bacterial resistance to antibiotics and heavy metals. We found that Typhimurium was the dominant S . enterica serovar, accounting for 68.3% (256/375) of the bird isolates. Furthermore, the proportions of the isolates identified as multi-antimicrobial resistant (multi-AMR: resistant to at least three antimicrobial classes) or multi-heavy metal resistant (multi-HMR: resistant to at least three heavy metals) were both 1.87% (7/375). Interestingly, all the multi-resistant S . enterica ( n = 12) were isolated from water birds or raptors; none of them was isolated from songbirds. Plasmid profiling demonstrated that 75% (9/12) of the multi-resistant strains carried resistance plasmids. Our study indicates that wild birds do not serve as important reservoirs of multi-resistant S . enterica strains. Nonetheless, continuous surveillance for bacterial resistance in wild birds is necessary because the multi-resistant isolates identified in this study also showed close genetic relatedness with those from humans and domestic animals.

Environmental Microbiology

Do pharmaceuticals, pathogens, and other organic waste water compounds persist when waste water is used for recharge?

A proof-of-concept experiment was devised to determine if pharmaceuticals and other organic waste water compounds (OWCs), as well as pathogens, found in treated effluent could be transported through a 2.4 m soil column and, thus, potentially reach ground water under recharge conditions similar to those in arid or semiarid climates. Treated effluent was applied at the top of the 2.4 m long, 32.5 cm diameter soil column over 23 days, Samples of the column inflow were collected from the effluent storage tank at the beginning (T begin ) and end (T end ) of the experiment, and a sample of the soil column drainage at the base of the column (B end ) was collected at the end of the experiment. Samples were analyzed for 131 OWCs including veterinary and human antibiotics, other prescription and nonprescription drugs, widely used household and industrial chemicals, and steroids and reproductive hormones, as well as the pathogens Salmonella and Legionella . Analytical results for the two effluent samples taken at the beginning (T begin ) and end (T end ) of the experiment indicate that the number of OWCs detected in the column inflow decreased by 25% (eight compounds) and the total concentration of OWCs decreased by 46% while the effluent was in the storage tank during the 23-day experiment. After percolating through the soil column, an additional 18 compounds detected in T end (67% of OWCs) were no longer detected in the effluent (B end ) and the total concentration of OWCs decreased by more than 70%. These compounds may have been subject to transformation (biotic and abiotic), adsorption, and (or) volatilization in the storage tank and during travel through the soil column. Eight compounds&mdash;carbamazapine; sulfamethoxazole; benzophenone; 5-methyl-1H-benzotriazole; N,N-diethyltoluamide; tributylphosphate; tri(2-chloroethyl) phosphate; and cholesterol&mdash;were detected in all three samples indicating they have the potential to reach ground water under recharge conditions similar to those in arid and semiarid climates. Results from real-time polymerase chain reactions demonstrated the presence of Legionella in all three samples. Salmonella was detected only in T begin , suggesting that the bacteria died off in the effluent storage tank over the period of the experiment. This proof-of-concept experiment demonstrates that, under recharge conditions similar to those in arid or semiarid climates, some pharmaceuticals, pathogens, and other OWCs can persist in treated effluent after soil-aquifer treatment.

Ground Water Monitoring and Remediation