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Transport and fate of microbial pathogens in agricultural settings

An understanding of the transport and survival of microbial pathogens (pathogens hereafter) in agricultural settings is needed to assess the risk of pathogen contamination to water and food resources, and to develop control strategies and treatment options. However, many knowledge gaps still remain in predicting the fate and transport of pathogens in runoff water, and then through the shallow vadose zone and groundwater. A number of transport pathways, processes, factors, and mathematical models often are needed to describe pathogen fate in agricultural settings. The level of complexity is dramatically enhanced by soil heterogeneity, as well as by temporal variability in temperature, water inputs, and pathogen sources. There is substantial variability in pathogen migration pathways, leading to changes in the dominant processes that control pathogen transport over different spatial and temporal scales. For example, intense rainfall events can generate runoff and preferential flow that can rapidly transport pathogens. Pathogens that survive for extended periods of time have a greatly enhanced probability of remaining viable when subjected to such rapid-transport events. Conversely, in dry seasons, pathogen transport depends more strongly on retention at diverse environmental surfaces controlled by a multitude of coupled physical, chemical, and microbiological factors. These interactions are incompletely characterized, leading to a lack of consensus on the proper mathematical framework to model pathogen transport even at the column scale. In addition, little is known about how to quantify transport and survival parameters at the scale of agricultural fields or watersheds. This review summarizes current conceptual and quantitative models for pathogen transport and fate in agricultural settings over a wide range of spatial and temporal scales. The authors also discuss the benefits that can be realized by improved modeling, and potential treatments to mitigate the risk of waterborne disease transmission.

Critical Reviews in Environmental Science and Tech

Population genetic structure of clinical and environmental isolates of Blastomyces dermatitidis based on 27 polymorphic microsatellite markers

Blastomyces dermatitidis , a thermally dimorphic fungus, is the etiologic agent of North American blastomycosis. Clinical presentation is varied, ranging from silent infections to fulminant respiratory disease and dissemination to skin and other sites. Exploration of the population genetic structure of B. dermatitidis would improve our knowledge regarding variation in virulence phenotypes, geographic distribution, and difference in host specificity. The objective of this study was to develop and test a panel of microsatellite markers to delineate the population genetic structure within a group of clinical and environmental isolates of B. dermatitidis . We developed 27 microsatellite markers and genotyped B. dermatitidis isolates from various hosts and environmental sources ( n =112). Assembly of a neighbor-joining tree of allele-sharing distance revealed two genetically distinct groups, separated by a deep node. Bayesian admixture analysis showed that two populations were statistically supported. Principal coordinate analysis also reinforced support for two genetic groups, with the primary axis explaining 61.41% of the genetic variability. Group 1 isolates average 1.8 alleles/locus, whereas group 2 isolates are highly polymorphic, averaging 8.2 alleles/locus. In this data set, alleles at three loci are unshared between the two groups and appear diagnostic. The mating type of individual isolates was determined by PCR. Both mating type-specific genes, the HMG and α-box domains, were represented in each of the genetic groups, with slightly more isolates having the HMG allele. One interpretation of this study is that the species currently designated B. dermatitidis includes a cryptic subspecies or perhaps a separate species.

Applied and Environmental Microbiology

Predictive models for Escherichia coli concentrations at inland lake beaches and relationship of model variables to pathogen detection

Predictive models, based on environmental and water quality variables, have been used to improve the timeliness and accuracy of recreational water quality assessments, but their effectiveness has not been studied in inland waters. Sampling at eight inland recreational lakes in Ohio was done in order to investigate using predictive models for Escherichia coli and to understand the links between E. coli concentrations, predictive variables, and pathogens. Based upon results from 21 beach sites, models were developed for 13 sites, and the most predictive variables were rainfall, wind direction and speed, turbidity, and water temperature. Models were not developed at sites where the E. coli standard was seldom exceeded. Models were validated at nine sites during an independent year. At three sites, the model resulted in increased correct responses, sensitivities, and specificities compared to use of the previous day's E. coli concentration (the current method). Drought conditions during the validation year precluded being able to adequately assess model performance at most of the other sites. Cryptosporidium , adenovirus, eaeA ( E. coli ), ipaH ( Shigella ), and spvC ( Salmonella ) were found in at least 20% of samples collected for pathogens at five sites. The presence or absence of the three bacterial genes was related to some of the model variables but was not consistently related to E. coli concentrations. Predictive models were not effective at all inland lake sites; however, their use at two lakes with high swimmer densities will provide better estimates of public health risk than current methods and will be a valuable resource for beach managers and the public.

Ohio

Understanding contaminants associated with mineral deposits

Interdisciplinary studies by the U.S. Geological Survey (USGS) have resulted in substantial progress in understanding the processes that control the release of metals and acidic water from inactive mines and mineralized areas, the transport of metals and acidic water to streams, and the fate and effect of metals and acidity on downstream ecosystems. The potential environmental effects associated with abandoned and inactive mines, resulting from the complex interaction of a variety of chemical and physical processes, is an area of study that is important to the USGS Mineral Resources Program. Understanding the processes contributing to the environmental effects of abandoned and inactive mines is also of interest to a wide range of stakeholders, including both those responsible for managing lands with historically mined areas and those responsible for anticipating environmental consequences of future mining operations. The recently completed (2007) USGS project entitled 'Process Studies of Contaminants Associated with Mineral Deposits' focused on abandoned and inactive mines and mineralized areas in the Rocky Mountains of Montana, Colorado, New Mexico, Utah, and Arizona, where there are thousands of abandoned mines. Results from these studies provide new information that advances our understanding of the physical and biogeochemical processes causing the mobilization, transport, reaction, and fate of potentially toxic elements (including aluminum, arsenic, cadmium, copper, iron, lead, and zinc) in mineralized near-surface systems and their effects on aquatic and riparian habitat. These interdisciplinary studies provide the basis for scientific decisionmaking and remedial action by local, State, and Federal agencies charged with minimizing the effects of potentially toxic elements on the environment. Current (2007) USGS research highlights the need to understand (1) the geologic sources of metals and acidity and the geochemical reactions that release them from their sources, (2) the pathways that facilitate transport from those sources, and (3) the processes that control the fate of the elements once released from the sources. Experts in the fields of economic geology, structural geology, mineralogy, geophysics, geochemistry, hydrology, ground-water modeling, microbiology, and toxicology came together for a series of studies that address these relationships on scales ranging from the microscopic to the watershed. This Circular presents results and highlights from the detailed, interdisciplinary studies that include investigations in both mining-affected areas and mineralized but unmined areas. The first section of the Circular describes laboratory and site-scale field investigations that primarily focus on mineralogic and biologic controls on the source and release of metals and acidity from mine-waste rock and hydrothermally altered areas. The second section describes a set of basin- to watershed-scale studies that not only investigate the source and release of metals and acidity but also the transport of these constituents away from the source areas. The third section is a summary of results from postremediation ecosystem monitoring. For more information on these and other project-related studies, please visit the project Web site at http://minerals.cr.usgs.gov/projects/contaminants/index.html. The Web site includes a complete bibliography and detailed descriptions of each interdisciplinary study.

Circular

Autoinducer 2: a concentration-dependent signal for mutualistic bacterial biofilm growth

4,5-dihydroxy-2,3-pentanedione (DPD), a product of the LuxS enzyme in the catabolism of S -ribosylhomocysteine, spontaneously cyclizes to form autoinducer 2 (AI-2). AI-2 is proposed to be a universal signal molecule mediating interspecies communication among bacteria. We show that mutualistic and abundant biofilm growth in flowing saliva of two human oral commensal bacteria, Actinomyces naeslundii T14V and Streptococcus oralis 34, is dependent upon production of AI-2 by S. oralis 34. A luxS mutant of S. oralis 34 was constructed which did not produce AI-2. Unlike wild-type dual-species biofilms, A. naeslundii T14V and an S. oralis 34 luxS mutant did not exhibit mutualism and generated only sparse biofilms which contained a 10-fold lower biomass of each species. Restoration of AI-2 levels by genetic or chemical (synthetic AI-2 in the form of DPD) complementation re-established the mutualistic growth and high biomass characteristic for the wild-type dual-species biofilm. Furthermore, an optimal concentration of DPD was determined, above and below which biofilm formation was suppressed. The optimal concentration was 100-fold lower than the detection limit of the currently accepted AI-2 assay. Thus, AI-2 acts as an interspecies signal and its concentration is critical for mutualism between two species of oral bacteria grown under conditions that are representative of the human oral cavity.

Molecular Microbiology

Evaluating the impacts of foreshore sand and birds on microbiological contamination at a freshwater beach

Beaches along the Great Lakes shorelines are important recreational and economic resources. However, contamination at the beaches can threaten their usage during the swimming season, potentially resulting in beach closures and/or advisories. Thus, understanding the dynamics that control nearshore water quality is integral to effective beach management. There have been significant improvements in this effort, including incorporating modeling (empirical, mechanistic) in recent years. Mechanistic modeling frameworks can contribute to this understanding of dynamics by determining sources and interactions that substantially impact fecal indicator bacteria concentrations, an index routinely used in water quality monitoring programs. To simulate E. coli concentrations at Jeorse Park beaches in southwest Lake Michigan, a coupled hydrodynamic and wave–current interaction model was developed that progressively added contaminant sources from river inputs, avian presence, bacteria–sediment interactions, and bacteria–sand–sediment interactions. Results indicated that riverine inputs affected E. coli concentrations at Jeorse Park beaches only marginally, while avian, shoreline sand, and sediment sources were much more substantial drivers of E. coli contamination at the beach. By including avian and riverine inputs, as well as bacteria–sand–sediment interactions at the beach, models can reasonably capture the variability in observed E. coli concentrations in nearshore water and bed sediments at Jeorse Park beaches. Consequently, it will be crucial to consider avian contamination sources and water-sand-sediment interactions in effective management of the beach for public health and as a recreational resource and to extend these findings to similar beaches affected by shoreline embayment.

Illlinois

Ideas and perspectives: A strategic assessment of methane and nitrous oxide measurements in the marine environment

>In the current era of rapid climate change, accurate characterization of climate-relevant gas dynamics – namely production, consumption, and net emissions – is required for all biomes, especially those ecosystems most susceptible to the impact of change. Marine environments include regions that act as net sources or sinks for numerous climate-active trace gases including methane (CH4) and nitrous oxide (N2O). The temporal and spatial distributions of CH4 and N2O are controlled by the interaction of complex biogeochemical and physical processes. To evaluate and quantify how these mechanisms affect marine CH4 and N2O cycling requires a combination of traditional scientific disciplines including oceanography, microbiology, and numerical modeling. Fundamental to these efforts is ensuring that the datasets produced by independent scientists are comparable and interoperable. Equally critical is transparent communication within the research community about the technical improvements required to increase our collective understanding of marine CH4 and N2O. A workshop sponsored by Ocean Carbon and Biogeochemistry (OCB) was organized to enhance dialogue and collaborations pertaining to marine CH4 and N2O. Here, we summarize the outcomes from the workshop to describe the challenges and opportunities for near-future CH4 and N2O research in the marine environment.

Biogeosciences

Microbial antimony biogeochemistry: Enzymes, regulation, and related metabolic pathways

Antimony (Sb) is a toxic metalloid that occurs widely at trace concentrations in soil, aquatic systems, and the atmosphere. Nowadays, with the development of its new industrial applications and the corresponding expansion of antimony mining activities, the phenomenon of antimony pollution has become an increasingly serious concern. In recent years, research interest in Sb has been growing and reflects a fundamental scientific concern regarding Sb in the environment. In this review, we summarize the recent research on bacterial antimony transformations, especially those regarding antimony uptake, efflux, antimonite oxidation, and antimonate reduction. We conclude that our current understanding of antimony biochemistry and biogeochemistry is roughly equivalent to where that of arsenic was some 20 years ago. This portends the possibility of future discoveries with regard to the ability of microorganisms to conserve energy for their growth from antimony redox reactions and the isolation of new species of “antimonotrophs.”

Applied and Environmental Microbiology

Vulnerability of high-latitude soil organic carbon in North America to disturbance

This synthesis addresses the vulnerability of the North American high-latitude soil organic carbon (SOC) pool to climate change. Disturbances caused by climate warming in arctic, subarctic, and boreal environments can result in significant redistribution of C among major reservoirs with potential global impacts. We divide the current northern high-latitude SOC pools into (1) near-surface soils where SOC is affected by seasonal freeze-thaw processes and changes in moisture status, and (2) deeper permafrost and peatland strata down to several tens of meters depth where SOC is usually not affected by short-term changes. We address key factors (permafrost, vegetation, hydrology, paleoenvironmental history) and processes (C input, storage, decomposition, and output) responsible for the formation of the large high-latitude SOC pool in North America and highlight how climate-related disturbances could alter this pool's character and size. Press disturbances of relatively slow but persistent nature such as top-down thawing of permafrost, and changes in hydrology, microbiological communities, pedological processes, and vegetation types, as well as pulse disturbances of relatively rapid and local nature such as wildfires and thermokarst, could substantially impact SOC stocks. Ongoing climate warming in the North American high-latitude region could result in crossing environmental thresholds, thereby accelerating press disturbances and increasingly triggering pulse disturbances and eventually affecting the C source/sink net character of northern high-latitude soils. Finally, we assess postdisturbance feedbacks, models, and predictions for the northern high-latitude SOC pool, and discuss data and research gaps to be addressed by future research.

Journal of Geophysical Research G: Biogeosciences

Beach monitoring criteria: reading the fine print

Beach monitoring programs aim to decrease swimming-related illnesses resulting from exposure to harmful microbes in recreational waters, while providing maximum beach access. Managers are advised by the U.S. EPA to estimate microbiological water quality based on a 5-day geometric mean of fecal indicator bacteria (FIB) concentrations or on a jurisdiction-specific single-sample maximum; however, most opt instead to apply a default single-sample maximum to ease application. We examined whether re-evaluation of the U.S. EPA ambient water quality criteria (AWQC) and the epidemiological studies on which they are based could increase public beach access without affecting presumed health risk. Single-sample maxima were calculated using historic monitoring data for 50 beaches along coastal Lake Michigan on various temporal and spatial groupings to assess flexibility in the application of the AWQC. No calculation on either scale was as low as the default maximum (235 CFU/100 mL) that managers typically use, indicating that current applications may be more conservative than the outlined AWQC. It was notable that beaches subject to point source FIB contamination had lower variation, highlighting the bias in the standards for these beaches. Until new water quality standards are promulgated, more site-specific application of the AWQC may benefit beach managers by allowing swimmers greater access to beaches. This issue will be an important consideration in addressing the forthcoming beach monitoring standards.

Environmental Science & Technology

Environmental Escherichia coli : Ecology and public health implications - A review

Escherichia coli is classified as a rod-shaped, Gram-negative bacterium in the family Enterobacteriaceae . The bacterium mainly inhabits the lower intestinal tract of warm-blooded animals, including humans, and is often discharged into the environment through feces or wastewater effluent. The presence of E. coli in environmental waters has long been considered as an indicator of recent fecal pollution. However, numerous recent studies have reported that some specific strains of E. coli can survive for long periods of time, and potentially reproduce, in extra-intestinal environments. This indicates that E. coli can be integrated into indigenous microbial communities in the environment. This naturalization phenomenon calls into question the reliability of E. coli as a fecal indicator bacterium (FIB). Recently, many studies reported that E. coli populations in the environment are affected by ambient environmental conditions affecting their long-term survival. Large-scale studies of population genetics provide the diversity and complexity of E. coli strains in various environments, affected by multiple environmental factors. This review examines the current knowledge on the ecology of E. coli strains in various environments in regards to its role as a FIB and as a naturalized member of indigenous microbial communities. Special emphasis is given on the growth of pathogenic E. coli in the environment, and the population genetics of environmental members of the genus Escherichia . The impact of environmental E. coli on water quality and public health is also discussed.

Journal of Applied Microbiology

Water quality and ecological risk assessment of intermittent streamflow through mining and urban areas of San Marcos River sub-basin, Mexico

Intermittent rivers are becoming more ecologically stressed worldwide. Flow cessation occurs naturally and spatiotemporally in these systems and anthropogenic activities such as wastewater discharges can have considerable impacts. Public entities mostly monitor water quality in permanent streams, leading to insufficient monitoring of intermittent streams and consequently to their potentially inadequate management.. This study analyzed spatiotemporal patterns of water quality and associated ecological risk through the quantification of physicochemical and microbiological pollutants in the intermittent river system of El Novillo and San Marcos in Northeast Mexico. Results showed that water quality varied geographically and seasonally. Based on national and international criteria, annual averages of water quality parameters analyzed suggested that streamflow in these river systems is of poor quality and poses high ecological risk to aquatic life. In the urban area, annual mean concentrations of Cd and Pb (0.14 and 0.4 mg/L) were 77- and 10-fold higher than their respective water quality criteria (<0.0018 and 0.04 mg/L). Statistically significant (q < 0.05) correlations were identified in concentrations of cyanide, Cd, Cu and Pb between wastewater seeping into the river and streamflow within the urban area. These observations highlight the unique sensitivity of intermittent urban streams to anthropogenic activities and may provide useful information to enhance current water management plans for the El Novillo-San Marcos River system for the protection of ecosystem integrity and human health.

El Novillo Canyon

A cross inoculation experiment reveals Ophidiomyces ophiodiicola and Nannizziopsis guarroi can each infect both snakes and lizards

Host range and specificity are key concepts in the study of infectious diseases. However, both concepts remain largely undefined for many influential pathogens, including many fungi within the Onygenales order. This order encompasses reptile-infecting genera ( Nannizziopsis , Ophidiomyces , and Paranannizziopsis ) formerly classified as the Chrysosporium anamorph of Nannizziopsis vriesii (CANV). The reported hosts of many of these fungi represent a narrow range of phylogenetically related animals, suggesting that many of these disease-causing fungi are host specific, but the true number of species affected by these pathogens is unknown. For example, to date, Nannizziopsis guarroi (the causative agent of yellow fungus disease) and Ophidiomyces ophiodiicola (the causative agent of snake fungal disease) have been documented only in lizards and snakes, respectively. In a 52-day reciprocal-infection experiment, we tested the ability of these two pathogens to infect currently unreported hosts, inoculating central bearded dragons ( Pogona vitticeps ) with O. ophiodiicola and corn snakes ( Pantherophis guttatus ) with N. guarroi . We confirmed infection by documenting both clinical signs and histopathological evidence of fungal infection. Our reciprocity experiment resulted in 100% of corn snakes and 60% of bearded dragons developing infections with N. guarroi and O. ophiodiicola , respectively, demonstrating that these fungal pathogens have a broader host range than previously thought and that hosts with cryptic infections may play a role in pathogen translocation and transmission.

Applied and Environmental Microbiology

USGS leads United States effort in Mallik Well

This winter, in the extremely cold, far reaches of the upper Northwest Territory of Canada, there is an international consortium of researchers participating in a program to study methane hydrates. The researchers are currently drilling a 1200 m-deep production research well through the permafrost. It is one of three wells located in the Mackenzie Delta, on the shore of the Beaufort Sea. Two observation wells were drilled adjacent to the main production test well earlier this year. Research objectives for the program focus on two themes: (1) the assessment of the production and properties of gas hydrates, and (2) an assessment of the stability of continental gas hydrates given warming trends predicted by climate change models. Of particular interest is the physical response of the gas hydrate to depressurization and thermal production stimulation. Cores are being taken from the well, and scientists hope to retrieve at least 200 m of core, including all the gas hydrate-rich intervals. Once cored, the samples are transported 200 kilometers over ice roads to Inuvik. Nearly 60 researchers are examining the cores for everything from geophysical parameters to microbiological analyses.

Fire in the Ice: NETL Methane Hydrate Newsletter

Understanding how microbiomes influence the systems they inhabit

Translating the ever-increasing wealth of information on microbiomes (environment, host, or built environment) to advance the understanding of system-level processes is proving to be an exceptional research challenge. One reason for this challenge is that relationships between characteristics of microbiomes and the system-level processes they influence are often evaluated in the absence of a robust conceptual framework and reported without elucidating the underlying causal mechanisms. The reliance on correlative approaches limits the potential to expand the inference of a single relationship to additional systems and advance the field. We propose that research focused on how microbiomes influence the systems they inhabit should work within a common framework and target known microbial processes that contribute to the system-level processes of interest. Here we identify three distinct categories of microbiome characteristics (microbial processes, microbial community properties, and microbial membership) and propose a framework to empirically link each of these categories to each other and the broader system level processes they affect. We posit that it is particularly important to distinguish microbial community properties that can be predicted from constituent taxa (community aggregated traits) from and those properties that are currently unable to be predicted from constituent taxa (emergent properties). Existing methods in microbial ecology can be applied to more explicitly elucidate properties within each of these categories and connect these three categories of microbial characteristics with each other. We view this proposed framework, gleaned from a breadth of research on environmental microbiomes and ecosystem processes, as a promising pathway with the potential to advance discovery and understanding across a broad range of microbiome science.

Nature Microbiology

SYBR green-based real-time reverse transcription-PCR for typing and subtyping of all hemagglutinin and neuraminidase genes of avian influenza viruses and comparison to standard serological subtyping tests

Continuing outbreaks of H5N1 highly pathogenic (HP) avian influenza virus (AIV) infections of wild birds and poultry worldwide emphasize the need for global surveillance of wild birds. To support the future surveillance activities, we developed a SYBR green-based, real-time reverse transcriptase PCR (rRT-PCR) for detecting nucleoprotein (NP) genes and subtyping 16 hemagglutinin (HA) and 9 neuraminidase (NA) genes simultaneously. Primers were improved by focusing on Eurasian or North American lineage genes; the number of mixed-base positions per primer was set to five or fewer, and the concentration of each primer set was optimized empirically. Also, 30 cycles of amplification of 1:10 dilutions of cDNAs from cultured viruses effectively reduced minor cross- or nonspecific reactions. Under these conditions, 346 HA and 345 NA genes of 349 AIVs were detected, with average sensitivities of NP, HA, and NA genes of 10 1.5 , 10 2.3 , and 10 3.1 50% egg infective doses, respectively. Utility of rRT-PCR for subtyping AIVs was compared with that of current standard serological tests by using 104 recent migratory duck virus isolates. As a result, all HA genes and 99% of the NA genes were genetically subtyped, while only 45% of HA genes and 74% of NA genes were serologically subtyped. Additionally, direct subtyping of AIVs in fecal samples was possible by 40 cycles of amplification: approximately 70% of HA and NA genes of NP gene-positive samples were successfully subtyped. This validation study indicates that rRT-PCR with optimized primers and reaction conditions is a powerful tool for subtyping varied AIVs in clinical and cultured samples.

Journal of Clinical Microbiology

Practical application of disease resistance: A brook trout fishery selected for resistance to furunculosis

Selection is unwittingly influenced by nuances of fish culturists and adaptation of fish to intensive culture within artificial environments. When pathogens are present, susceptible individuals may be selectively eliminated from a population, accounting for inter-specific and intra-specific differences in susceptibility to disease. Biologists often intensify such selection to enhance resistance. Particularly important to this discourse is the selective breeding program initiated by the New York State Department of Environmental Conservation (Rome, NY, USA) to produce brown trout ( Salmo salar ) and brook trout ( Salvelinus fontinalis ) that are resistant to Aeromonas salmonicida . This program, specifically as it relates to brook trout, is reviewed throughout the current manuscript. In addition, we present a comparison between the performance of the Rome strain of brook trout to that of the Owhi strain of brook trout during a 17-month production cycle at the Ed Weed Fish Culture Station (Grand Isle, VT, USA) and after stocking into Vermont waters. The Owhi strain is used in recreational fisheries because it has good post-stocking survival, but these fish are sensitive to furunculosis. Our data indicated that the Rome strain had a food conversion rate of 1.3 that was slightly less than the 1.1 conversion rate of the Owhi brook trout. The growth rate measured as Monthly Temperature Units per inch (MTU/in.) among Rome brook trout (24.9 MTU/in.) was better than the Owhi brook trout (29.5 MTU/in.). Both the average length (23.6 cm) and weight (159.7 g) of individual Rome brook trout were superior to those of the Owhi strain (20.4 cm and 78.2 g, respectively). Just prior to stocking, survival within the hatchery was 84% among the Rome trout, but only 51% among the Owhi brook trout and mortality was attributed to a persistent epizootic of furunculosis. Microbiological assays indicated that the prevalence of A. salmonicida in dermal mucus attained equivalent levels within both strains of fish at different phases of the production cycle. The level of systemic infection, however, was regulated to a much greater extent by the Rome strain of brook trout throughout production. Our results indicated that the Rome brook trout strain satisfies management objectives for establishing an effective recreational brook trout fishery. Use of this strain also reduces management issues that complicate production when A. salmonicida is enzootic.

Vermont

Diversity, composition, and geographical distribution of microbial communities in California salt marsh sediments

The Pacific Estuarine Ecosystem Indicators Research Consortium seeks to develop bioindicators of toxicant-induced stress and bioavailability for wetland biota. Within this framework, the effects of environmental and pollutant variables on microbial communities were studied at different spatial scales over a 2-year period. Six salt marshes along the California coastline were characterized using phospholipid fatty acid (PLFA) analysis and terminal restriction fragment length polymorphism (TRFLP) analysis. Additionally, 27 metals, six currently used pesticides, total polychlorinated biphenyls and polycyclic aromatic hydrocarbons, chlordanes, nonachlors, dichlorodiphenyldichloroethane, and dichlorodiphenyldichloroethylene were analyzed. Sampling was performed over large (between salt marshes), medium (stations within a marsh), and small (different channel depths) spatial scales. Regression and ordination analysis suggested that the spatial variation in microbial communities exceeded the variation attributable to pollutants. PLFA analysis and TRFLP canonical correspondence analysis (CCA) explained 74 and 43% of the variation, respectively, and both methods attributed 34% of the variation to tidal cycles, marsh, year, and latitude. After accounting for spatial variation using partial CCA, we found that metals had a greater effect on microbial community composition than organic pollutants had. Organic carbon and nitrogen contents were positively correlated with PLFA biomass, whereas total metal concentrations were positively correlated with biomass and diversity. Higher concentrations of heavy metals were negatively correlated with branched PLFAs and positively correlated with methyl- and cyclo-substituted PLFAs. The strong relationships observed between pollutant concentrations and some of the microbial indicators indicated the potential for using microbial community analyses in assessments of the ecosystem health of salt marshes. Copyright ?? 2006, American Society for Microbiology. All Rights Reserved.

Applied and Environmental Microbiology