Search USGSSearch

SEARCH · Search USGS

Results for “Environmental DNA”

Search indexed USGS publications on groundwater, aquifers, geologic maps, mineral resources and earthquakes. Explore source records by subject and place.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 253 records · Page 14Linked to original sources

Estrogen receptor 1 expression and methylation of Esr1 promoter in mouse fetal prostate mesenchymal cells induced by gestational exposure to bisphenol A or ethinylestradiol

Fetal/neonatal environmental estrogen exposures alter developmental programing of the prostate gland causing onset of diseases later in life. We have previously shown in vitro that exposures to 17β-estradiol (E2) and the endocrine disrupting chemical bisphenol A, at concentrations relevant to human exposure, cause an elevation of estrogen receptor α ( Esr1 ) mRNA in primary cultures of fetal mouse prostate mesenchymal cells; a similar result was observed in the fetal rat urogenital sinus. Effects of these chemicals on prostate mesenchyme in vivo are not well understood. Here we show effects in mice of fetal exposure to the estrogenic drug in mixed oral contraceptives, 17α-ethinylestradiol (EE2), at a concentration of EE2 encountered by human embryos/fetuses whose mothers become pregnant while on EE2-containing oral contraceptives, or bisphenol A at a concentration relevant to exposures observed in human fetuses in vivo . Expression of Esr1 was elevated by bisphenol A or EE2 exposures, which decreased the global expression of DNA methyltransferase 3A ( Dnmt3a ), while methylation of Esr1 promoter was significantly increased. These results show that exposures to the environmental estrogen bisphenol A and drug EE2 cause transcriptional and epigenetic alterations to expression of estrogen receptors in developing prostate mesenchyme in vivo .

Environmental Epigenetics

The role of neutral and adaptive genomic variation in population diversification and speciation in two ground squirrel species of conservation concern

Understanding the neutral (demographic) and adaptive processes leading to the differentiation of species and populations is a critical component of evolutionary and conservation biology. In this context, recently diverged taxa represent a unique opportunity to study the process of genetic differentiation. Northern and southern Idaho ground squirrels ( Urocitellus brunneus —NIDGS, and U . endemicus —SIDGS, respectively) are a recently diverged pair of sister species that have undergone dramatic declines in the last 50 years and are currently found in metapopulations across restricted spatial areas with distinct environmental pressures. Here we genotyped single-nucleotide polymorphisms (SNPs) from buccal swabs with restriction site-associated DNA sequencing (RADseq). With these data we evaluated neutral genetic structure at both the inter- and intraspecific level, and identified putatively adaptive SNPs using population structure outlier detection and genotype–environment association (GEA) analyses. At the interspecific level, we detected a clear separation between NIDGS and SIDGS, and evidence for adaptive differentiation putatively linked to torpor patterns. At the intraspecific level, we found evidence of both neutral and adaptive differentiation. For NIDGS, elevation appears to be the main driver of adaptive differentiation, while neutral variation patterns match and expand information on the low connectivity between some populations identified in previous studies using microsatellite markers. For SIDGS, neutral substructure generally reflected natural geographical barriers, while adaptive variation reflected differences in land cover and temperature, as well as elevation. These results clearly highlight the roles of neutral and adaptive processes for understanding the complexity of the processes leading to species and population differentiation, which can have important conservation implications in susceptible and threatened species.

Idaho

Cytochrome b sequences in black-crowned night-herons (Nycticorax nycticorax) from heronries exposed to genotoxic contaminants

DNA sequence analysis of a 215 base-pair region of the mitochondrial cytochrome b gene was used to examine genetic variation and search for evidence of an increased mutation rate in black-crowned night-herons. We examined five populations exposed to environmental contamination (primarily PAHs and PCBs) and one reference population from the eastern U.S. There was no evidence of a high mutation rate even within populations previously shown to exhibit increased variation in DNA content among somatic cells as a result of petroleum exposure. Three haplotypes were observed among 99 individuals. The low level of variability could be evidence for a genetic bottleneck, or that cytochrome b is too conservative for use in population genetic studies of this species. With the exception of one population from Louisiana, pair-wise Phi st estimates were very low, indicative of little population structure and potentially high rates of effective migration among populations.

Ecotoxicology

Museum metabarcoding: a novel method revealing gut helminth communities of small mammals across space and time

Natural history collections spanning multiple decades provide fundamental historical baselines to measure and understand changing biodiversity. New technologies such as next generation DNA sequencing (NGS) have considerably increased the potential of museum specimens to address significant questions regarding the impact of environmental changes on host and parasite/pathogen dynamics. We developed a new technique to identify intestinal helminth parasites and applied it to shrews (Eulipotyphla: Soricidae) because they are ubiquitous, occupy diverse habitats, and host a diverse and abundant parasite fauna. Notably, we included museum specimens preserved in various ways to explore the efficacy of using metabarcoding analyses that may enable identification of helminth symbiont communities from historical archives. We successfully sequenced the parasite communities (using 12S mtDNA, 16S mtDNA, 28S rDNA) of 23 whole gastrointestinal (GI) tracts. All GI tracts were obtained from the Museum of Southwestern Biology (MSB), USA, and from recent field collections, varying both in time since fixation (ranging from 4 months to 16 years) and preservation method (70% or 95% ethanol stored at room temperature, or flash frozen in liquid nitrogen and stored at -80°C). Our proof of concept demonstrates the feasibility of applying NGS techniques to authoritatively identify the parasite/pathogen communities within whole GI tracts from museum specimens of varying age and fixation, and the value of future preservation of host-associated whole GI tracts in public research archives. This powerful approach facilitates future comparative examinations of the distributions and interactions among multiple associated groups of organisms through time and space.

International Journal for Parasitology

Urbanization and host relatedness shape virome composition in a widespread, generalist carnivore

Urban wildlife species have the potential to serve as links in disease transmission between wildlife, humans and domestic animals at the wildland–urban interface (WUI), contributing to both sustained cross-species transmission of pathogens and the emergence of diseases in susceptible populations. However, the relative roles of host and environmental factors in shaping the composition of pathogen communities in urban wildlife is understudied. In this study, we integrated DNA and RNA virome data with host genomic and GPS datasets to investigate factors shaping virome composition in bobcats ( Lynx rufus ) at the WUI in the Tucson Mountains, Arizona, USA. Using a hybrid-capture approach for 31 scats and 17 buccal swabs, we identified multiple viruses that could affect carnivore health at the WUI, including canine parvovirus, feline astrovirus, Felis catus papillomaviruses 2 and 3 and Lyon-IARC polyomavirus. Models of virome composition and distribution of viral taxa indicated contributions of host genetic relatedness and factors relating to urbanisation (such as percentages of urban land cover, road and building densities and distances to roads). Genetic associations with virome compositions were particularly influenced by females. While females exhibit significant isolation by distance, partial Mantel tests revealed a significant correlation between beta diversity and host genetic distance in females only. To our knowledge, this study represents the first assessment of factors shaping virome composition in a wild felid. Our finding of known feline and canine pathogens in bobcats underscores the potential of the WUI to facilitate cross-species transmission between wild and domestic animals.

Arizona

Genomic and environmental influences on resilience in a cold-water fish near the edge of its range

Small, isolated populations present a challenge for conservation. The dueling effects of selection and drift in a limited pool of genetic diversity make the responses of small populations to environmental perturbations erratic and difficult to predict. This is particularly true at the edge of a species range, where populations often persist at the limits of their environmental tolerances. Populations of cisco, Coregonus artedi , in inland lakes have experienced numerous extirpations along the southern edge of their range in recent decades, which are thought to result from environmental degradation and loss of cold, well-oxygenated habitat as lakes warm. Yet, cisco extirpations do not show a clear latitudinal pattern, suggesting that local environmental factors and potentially local adaptation may influence resilience. Here, we used genomic tools to investigate the nature of this pattern of resilience. We used restriction site-associated DNA capture (Rapture) sequencing to survey genomic diversity and differentiation in southern inland lake cisco populations and compared the frequency of deleterious mutations that potentially influence fitness across lakes. We also examined haplotype diversity in a region of the major histocompatibility complex involved in stress and immune system response. We correlated these metrics to spatial and environmental factors including latitude, lake size, and measures of oxythermal habitat and found significant relationships between genetic metrics and broad and local factors. High levels of genetic differentiation among populations were punctuated by a phylogeographic break and residual patterns of isolation-by-distance. Although the prevalence of deleterious mutations and inbreeding coefficients was significantly correlated with latitude, neutral and non-neutral genetic diversity were most strongly correlated with lake surface area. Notably, differences among lakes in the availability of estimated oxythermal habitat left no clear population genomic signature. Our results shed light on the complex dynamics influencing these isolated populations and provide valuable information for their conservation.

Wisconsin

DNA metabarcoding and video camera collars yield different inferences about the summer diet of an arctic ungulate

The diets of wild ungulates are a foundational component of their ecology, influencing their behavior, body condition, and demography. With changing environmental conditions, there is a significant need to identify important forage items for ungulates, but this has often proved challenging. Declines in several barren-ground caribou herds across the North American Arctic have raised concerns about the influence of climate change on caribou forage conditions. Shifts in plant phenology, biomass, quality, and composition may be influencing caribou diets and subsequently affecting their body condition and demographic rates. Although forage is a primary driver of barren-ground caribou behavior and population dynamics, there is limited recent information about the specific foods they consume, and uncertainty about appropriate methods for identifying those foods. Investigators are increasingly using fecal DNA metabarcoding and video camera collars to assess ungulate diets, but comparative studies of these approaches are lacking. To examine the summer diets of barren-ground caribou, we used both fecal metabarcoding and video camera collars to identify forage used by the Porcupine caribou herd, which spans the Alaska–Yukon border. In 2021, we sampled the diets of adult females by collecting fecal samples and observing collar videos during 4 sampling occasions. We found that caribou consumed very specific forage items, and those items varied markedly across the growing season. Caribou predominantly consumed graminoids and lichens during early summer, and shrubs and forbs later in the season. Metabarcoding and video data provided significantly different estimates of diet for all taxonomic levels we evaluated, and inferences from the two approaches were often disparate. Metabarcoding failed to detect some items frequently consumed in videos, such as lichens, and indicated high use of other items rarely consumed, such as mosses. We found that video data provided greater taxonomic diversity and resolution for vascular plants and lichens, and more closely aligned with past research and biological expectations than data from fecal metabarcoding. Additional research is needed to be able to use these methods to identify the biomass of different forage items consumed.

Alaska, Yukon

Water Quality, Cyanobacteria, and Environmental Factors and Their Relations to Microcystin Concentrations for Use in Predictive Models at Ohio Lake Erie and Inland Lake Recreational Sites, 2013-14

Harmful cyanobacterial &ldquo;algal&rdquo; blooms (cyanoHABs) and associated toxins, such as microcystin, are a major water-quality issue for Lake Erie and inland lakes in Ohio. Predicting when and where a bloom may occur is important to protect the public that uses and consumes a water resource; however, predictions are complicated and likely site specific because of the many factors affecting toxin production. Monitoring for a variety of environmental and water-quality factors, for concentrations of cyanobacteria by molecular methods, and for algal pigments such as chlorophyll and phycocyanin by using optical sensors may provide data that can be used to predict the occurrence of cyanoHABs. To test these monitoring approaches, water-quality samples were collected at Ohio recreational sites during May&ndash;November in 2013 and 2014. In 2013, samples were collected monthly at eight sites at eight lakes to facilitate an initial assessment and select sites for more intensive sampling during 2014. In 2014, samples were collected approximately weekly at five sites at three lakes. Physical water-quality parameters were measured at the time of sampling. Composite samples were preserved and analyzed for dissolved and total nutrients, toxins, phytoplankton abundance and biovolume, and cyanobacterial genes by molecular methods. Molecular assays were done to enumerate (1) general cyanobacteria, (2) general Microcystis and Dolichospermum ( Anabaena ), (3) mcyE genes for Microcystis , Dolichospermum ( Anabaena ), and Planktothrix targeting deoxyribonucleic acid (DNA), and (4) mcyE transcripts for Microcystis , Dolichospermum ( Anabaena ), and Planktothrix targeting ribonucleic acid (RNA).The DNA assays for the mcyE gene provide data on cyanobacteria that have the potential to produce microcystin, whereas the RNA assays provide data on cyanobacteria that are actively transcribing the toxin gene. Environmental data were obtained from available online sources. Quality-control (QC) samples were collected and analyzed for all constituents to characterize bias and variability; however, QC data for molecular assays were examined in more detail than for the other constituents. The QC data for molecular assays suggested that sampling variability and qPCR variability were small in comparison with the combined variability associated with sample filtering, extraction and purification, and the matrix itself. A total of 46 water-quality samples were collected during 2013 at 8 beach sites&mdash;Buck Creek, Buckeye Crystal, Deer Creek, Harsha Main, Maumee Bay State Park (MBSP) Inland (negative control site), MBSP Lake Erie, Port Clinton, and Sandusky Bay. Microcystin was detected in 67&ndash;100 percent of samples at all sites except for MBSP Inland, where microcystin was detected in only 20 percent of samples. Microcystin concentrations ranged from <0.10 to 48 micrograms per liter (&micro;/L), with the widest range found at MBSP Lake Erie and the highest concentrations found at Buckeye Crystal. Saxitoxin was detected in five samples, and cylindrospermopsin was not detected in any samples. A total of 65 water-quality samples were collected during 2014 at 5 sites on 3 lakes&mdash;Buckeye Fairfield and Onion Island, Harsha Main and Campers, and MBSP Lake Erie beach. Four of the sites were bathing beaches and one site, Onion Island, was an offshore boater swim area. Concentrations of microcystin ranged from <0.10 to 240 &micro;/L and, as in 2013, the widest range was found at MBSP Lake Erie. At Buckeye Lake, microcystin concentrations were consistently high (greater than 20 &micro;/L), ranging from 23 to 81 &micro;/L. At Harsha Main and Campers, microcystin concentrations ranged from <0.10 to 15 &micro;/L. Saxitoxin was detected in four samples collected at MBSP Lake Erie. Throughout the 2014 season, the cyanobacterial community, as determined by molecular and microscopy methods, and the dominance associated with the highest microcystin concentrations were unique to individual lakes. At Buckeye Lake, Planktothrix dominated the cyanobacterial community throughout the season and Planktothrix DNA and RNA were found in 100 percent of samples; Microcystis mcyE DNA was found in low concentrations. At Harsha Lake, Dolichospermum and Microcystis were a substantial percentage of the community from late May through August, and the highest microcystin concentrations occurred in June and July. At MBSP Lake Erie, Microcystis generally dominated from mid-July through early November, and the highest microcystin concentrations occurred in August. Spearman&rsquo;s correlation coefficient (rho) was computed to determine the relations between environmental and water-quality factors and microcystin concentrations at four sites&mdash;Buckeye Fairfield, Buckeye Onion Island, Harsha Main, and MBSP Lake Erie. Factors were evaluated for use as potential independent variables in two types of predictive models&mdash;daily and long-term models. Easily or continuously measured water-quality factors and available environmental data are used for daily predictions that do not require a site visit. Data from factors used in daily predictions and results from samples collected and analyzed in a laboratory are used for long-term predictions (a few days to several weeks). A few statistically significant correlations (p &le; 0.05) between microcystin concentrations and factors for both daily and long-term predictions were found at Buckeye Onion Island, and many were found at Harsha Main and MBSP Lake Erie. There were only a few statistically significant factors for daily predictions at Buckeye Fairfield, likely because of the lack of variability in microcystin concentrations. Among factors for daily predictions, phycocyanin had the highest Spearman&rsquo;s correlation to microcystin concentrations (rho = 0.79 to 0.93) at all sites except for Buckeye Fairfield. Turbidity, pH, algae category, and Secchi depth were significantly correlated to microcystin concentrations at Harsha Main and MBSP Lake Erie. Algae categories were observational categories from 0 (none) to 4 (extreme). Several discharge variables (Maumee River at Waterville, river mouth is approximately 3.5 miles from the beach) at MBSP Lake Erie were promising environmental factors for daily predictions. In addition to discrete water-quality measurements recorded at Harsha Main at the time of sampling, many manipulated measurements (factors derived from mathematical manipulation of time-series data) available from a nearby continuous monitor were strongly correlated to microcystin concentrations; the highest correlation was found for the relation between microcystin concentrations and the antecedent 7-day average phycocyanin (rho = 0.98). For long-term predictions, the most highly correlated molecular assays were Planktothrix mcyE DNA at Buckeye Onion Island and Microcystis mcyE DNA at Harsha Main and MBSP Lake Erie. Concentrations of several nutrient constituents were significantly correlated to microcystin concentrations including total nitrogen at Buckeye Onion Island, ammonia and nitrate plus nitrite (both negatively correlated) at Harsha Main and MBSP Lake Erie, and total phosphorus at MBSP Lake Erie. The results of this study showed that water-quality and environmental variables are promising for use in site-specific daily or long-term predictive models. In order to develop more accurate models to predict toxin concentrations at freshwater lake sites, data need to be collected more frequently and for consecutive days in future studies.

Ohio

Evaluation of two spike-and-recovery controls for assessment of extraction efficiency in microbial source tracking studies

Quantitative PCR (qPCR), applied to complex environmental samples such as water, wastewater, and feces, is susceptible to methodological and sample related biases. In this study, we evaluated two exogenous DNA spike-and-recovery controls as proxies for recovery efficiency of Bacteroidales 16S rDNA gene sequences (AllBac and qHF183) that are used for microbial source tracking (MST) in river water. Two controls-(1) the plant pathogen Pantoea stewartii, carrying the chromosomal target gene cpsD, and (2) Escherichia coli, carrying the plasmid-borne target gene DsRed2-were added to raw water samples immediately prior to concentration and DNA extraction for qPCR. When applied to samples processed in replicate, recovery of each control was positively correlated with the observed concentration of each MST marker. Adjustment of MST marker concentrations according to recovery efficiency reduced variability in replicate analyses when consistent processing and extraction methodologies were applied. Although the effects of this procedure on accuracy could not be tested due to uncertainties in control DNA concentrations, the observed reduction in variability should improve the strength of statistical comparisons. These findings suggest that either of the tested spike-and-recovery controls can be useful to measure efficiency of extraction and recovery in routine laboratory processing. ?? 2009 Elsevier Ltd.

Water Research

Thirteen novel ideas and underutilized resources to support progress towards a range-wide American eel stock assessment

A robust assessment of the American eel ( Anguilla rostrata ) stock, required to guide conservation efforts, is challenged by the species’ vast range, high variability in demographic parameters and data inadequacies. Novel ideas and underutilised resources that may assist both analytic assessments and spatially oriented modelling include (1) species and environmental databases; (2) mining of data from scattered sources; (3) infilling of data gaps by spatial analysis; (4) age estimation from measurements of DNA methylation; evaluation of eel abundance by (5) larval, (6) glass-bottom boat, (7) net enclosure and (8) eDNA surveys; (9) accounting for dam-induced habitat increases in eel watercourse modelling; (10) spatially oriented modelling with and without temporal components; (11) geographically nested modelling of glass eel recruitment; (12) spawner per recruit modelling and (13) life cycle modelling to examine larval allocation effects. Eel biologists are too few to gather the required assessment data across all of the species’ range. Public posting of electrofishing and eDNA metabarcoding data sets and the use of machine learning techniques to comprehensively inventory small dams will help meet some data needs. These approaches address only a small proportion of the assessment challenges that face American eels. Worldwide collaboration amongst Anguilla scientists is a key enabler of progress towards stock assessment goals.

North America, South America

Exploration of the 2016 Yellowstone River fish kill and proliferative kidney disease in wild fish populations

Proliferative kidney disease (PKD) is an emerging disease that recently resulted in a large mortality event of salmonids in the Yellowstone River (Montana, USA). Total PKD fish mortalities in the Yellowstone River were estimated in the tens of thousands, which resulted in a multi‐week river closure and an estimated economic loss of US$500,000. This event shocked scientists, managers, and the public, as this was the first occurrence of the disease in the Yellowstone River, the only reported occurrence of the disease in Montana in the past 25 yr, and arguably the largest wild PKD fish kill in the world. To understand why the Yellowstone River fish kill occurred, we used molecular and historical data to evaluate evidence for several hypotheses: Was the causative parasite Tetracapsuloides bryosalmonae a novel invader, was the fish kill associated with a unique parasite strain, and/or was the outbreak caused by unprecedented environmental conditions? We found that T. bryosalmonae is widely distributed in Montana and have documented occurrence of this parasite in archived fish collected in the Yellowstone River prior to the fish kill. T. bryosalmonae had minimal phylogeographic population structure, as the DNA of parasites sampled from the Yellowstone River and distant water bodies were very similar. These results suggest that T. bryosalmonae could be endemic in Montana. Due to data limitations, we could not reject the hypothesis that the fish kill was caused by a novel and more virulent genetic strain of the parasite. Finally, we found that single‐year environmental conditions are insufficient to explain the cause of the 2016 Yellowstone River PKD outbreak. Other regional rivers where we documented T. bryosalmonae had similar or even more extreme conditions than the Yellowstone River and similar or more extreme conditions have occurred in the Yellowstone River in the recent past, yet mass PKD mortalities have not been documented in either instance. We conclude by placing these results and unresolved hypotheses into the broader context of international research on T. bryosalmonae and PKD, which strongly suggests that a better understanding of bryozoans, the primary host of T. bryosalmonae , is required for better ecosystem understanding.

Montana

Comparative assessment of a restored and natural wetland using 13C-DNA SIP reveals a higher potential for methane production in the restored wetland

Wetlands are the largest natural source of methane (CH 4 ), a potent greenhouse gas produced by methanogens. Methanogenesis rates are controlled by environmental factors such as redox potential, temperature, and carbon and electron acceptor availability and are presumably dependent on the composition of the active methanogen community. We collected intact soil cores from a restored and natural freshwater depressional wetland on Maryland’s Delmarva Peninsula (USA) to assess the effects of wetland restoration and redox shifts on microbial processes. Intact soil cores were incubated under either saturated (anoxic) or unsaturated (oxic) conditions and amended with 13 C-acetate for quantitative stable isotope probing (qSIP) of the 16S rRNA gene. Restored wetland cores supported a distinct community of methanogens compared to natural cores, and acetoclastic methanogens putatively identified in the genus Methanosarcina were among the most abundant taxa in restored anoxic and oxic cores. The active microbial communities in the restored wetland cores were also distinguished by the unique presence of facultatively anaerobic bacteria belonging to the orders Firmicutes and Bacteroidetes . In natural wetland incubations, methanogen populations were not among the most abundant taxa, and these communities were instead distinguished by the unique presence of aerobic bacteria in the phyla Acidobacteria , Actinobacteria , and class Alphaproteobacteria . Iron-reducing bacteria, in the genus Geobacter , were active across all redox conditions in both the restored and the natural cores, except the natural oxic–anoxic condition. These findings suggest an overall higher potential for methanogenesis in the restored wetland site compared to the natural wetland site, even when there is evidence of Fe reduction.

Maryland

A new DNA extraction method (HV-CTAB-PCI) for amplification of nuclear markers from open ocean-retrieved faeces of an herbivorous marine mammal, the dugong

Non-invasively collected faecal samples are an alternative source of DNA to tissue samples, that may be used in genetic studies of wildlife when direct sampling of animals is difficult. Although several faecal DNA extraction methods exist, their efficacy varies between species. Previous attempts to amplify mitochondrial DNA (mtDNA) markers from faeces of wild dugongs ( Dugong dugon ) have met with limited success and nuclear markers (microsatellites) have been unsuccessful. This study aimed to establish a tool for sampling both mtDNA and nuclear DNA (nDNA) from dugong faeces by modifying approaches used in studies of other large herbivores. First, a streamlined, cost-effective DNA extraction method that enabled the amplification of both mitochondrial and nuclear markers from large quantities of dugong faeces was developed. Faecal DNA extracted using a new ‘High Volume- Cetyltrimethyl Ammonium Bromide- Phenol-Chloroform-Isoamyl Alcohol’ (HV-CTAB-PCI) method was found to achieve comparable amplification results to extraction of DNA from dugong skin. As most prevailing practices advocate sampling from the outer surface of a stool to maximise capture of sloughed intestinal cells, this study compared amplification success of mtDNA between the outer and inner layers of faeces, but no difference in amplification was found. Assessment of the impacts of faecal age or degradation on extraction, however, demonstrated that fresher faeces with shorter duration of environmental (seawater) exposure amplified both markers better than eroded scats. Using the HV-CTAB-PCI method, nuclear markers were successfully amplified for the first time from dugong faeces. The successful amplification of single nucleotide polymorphism (SNP) markers represents a proof-of-concept showing that DNA from dugong faeces can potentially be utilised in population genetic studies. This novel DNA extraction protocol offers a new tool that will facilitate genetic studies of dugongs and other large and cryptic marine herbivores in remote locations.

Queensland

Cell proliferation detected with flow cytometric cell cycle analysis and immunohistochemical detection of Proliferating Cell Nuclear Antigen (PCNA) from somatic tissues of Eastern Oysters, Crassostrea virginica

Two novel biomarker of response assays were developed and compared for use with bivalves. Bivalve mollusks are often used as bioindicators to monitor contaminant body burdens and are employed globally in pollution monitoring and as sentinels of environmental quality. The prevalence of proliferating cells in tissues of the eastern oyster, Crassostrea virginica , (n = 10) was investigated by using immunohistochemical staining of proliferating cell nuclear antigen (PCNA) and by flow cytometric analysis of DNA in cell cycle phases. The PCNA protein is highly conserved and central to DNA synthesis in dividing cells. This protein was detected by using a commercial antibody in the histological staining of gills, mantle, digestive gland, stomach, and labial palps. Cells obtained from these same organs and from the hearts of the oysters were used for flow cytometry (FCM). The proportions of nuclei in the S plus G2/M and G2/M fractions were compared with percentages of PCNA-positive cells scored through microscopy. Proliferation responses generated by microscopy and FCM (S plus G2/M) were equal for digestive gland, gills, stomach, and mantle. The proliferation response in labial palps was significantly higher by PCNA microscopy than by DNA FCM (S plus G2/M). For PCNA microscopy results, a significantly higher proliferation response was noted both for labial palps as compared to gills and mantle and for stomach and digestive gland as compared to gills (P < 0.005). For DNA FCM results, the percentages of proliferating cells in G2M fractions were significantly higher for labial palps and heart when compared to gills, mantle, and digestive gland. By FCM (S plus G2M), the percentages of proliferating cells followed a similar trend, although no significant differences were found between organs. The choice of a relatively highly proliferative tissue, such as labial palps as determined by this study, has utility for developing a cell line, for examining proliferation as a biomarker due to stressor impacts in feral oysters, and for detecting impacts of compounds on proliferative mechanisms.

Environmental Bioindicators

Sperm quality biomarkers complement reproductive and endocrine parameters in investigating environmental contaminants in common carp (Cyprinus carpio) from the Lake Mead National Recreation Area

Lake Mead National Recreational Area (LMNRA) serves as critical habitat for several federally listed species and supplies water for municipal, domestic, and agricultural use in the Southwestern U.S. Contaminant sources and concentrations vary among the sub-basins within LMNRA. To investigate whether exposure to environmental contaminants is associated with alterations in male common carp ( Cyprinus carpio ) gamete quality and endocrine- and reproductive parameters, data were collected among sub-basins over 7 years (1999–2006). Endpoints included sperm quality parameters of motility , viability, mitochondrial membrane potential, count, morphology, and DNA fragmentation; plasma components were vitellogenin (VTG), 17ß-estradiol, 11-keto-testosterone, triiodothyronine, and thyroxine. Fish condition factor, gonadosomatic index, and gonadal histology parameters were also measured. Diminished biomarker effects were noted in 2006, and sub-basin differences were indicated by the irregular occurrences of contaminants and by several associations between chemicals (e.g., polychlorinated biphenyls, hexachlorobenzene , galaxolide, and methyl triclosan) and biomarkers (e.g., plasma thyroxine, sperm motility and DNA fragmentation). By 2006, sex steroid hormone and VTG levels decreased with subsequent reduced endocrine disrupting effects. The sperm quality bioassays developed and applied with carp complemented endocrine and reproductive data, and can be adapted for use with other species.

Lake Mead National Recreation Area

Dispersal hazards of Pseudogymnoascus destructans by bats and human activity at hibernacula in summer

Bats occupying hibernacula during summer are exposed to Pseudogymnoascus destructans ( Pd ), the causative agent of white-nose syndrome (WNS), and may contribute to its dispersal. Furthermore, equipment and clothing exposed to cave environments are a potential source for human-assisted spread of Pd . To explore dispersal hazards for Pd during the nonhibernal season, we tested samples that were collected from bats, the environment, and equipment at hibernacula in the eastern US between 18 July–22 August 2012. Study sites included six hibernacula known to harbor bats with Pd with varying winter-count impacts from WNS and two hibernacula (control sites) without prior history of WNS. Nucleic acid from Pd was detected from wing-skin swabs or guano from 40 of 617 bats (7% prevalence), including males and females of five species at five sites where WNS had previously been confirmed as well as from one control site. Analysis of guano collected during summer demonstrated a higher apparent prevalence of Pd among bats (17%, 37/223) than did analysis of wing-skin swabs (1%, 4/617). Viable Pd cultured from wing skin (2%, 1/56) and low recapture rates at all sites suggested bats harboring Pd during summer could contribute to pathogen dispersal. Additionally, Pd DNA was detected on clothing and trapping equipment used inside and near hibernacula, and Pd was detected in sediment more readily than in swabs of hibernaculum walls. Statistically significant differences in environmental abundance of Pd were not detected among sites, but prevalence of Pd differed between sites and among bat species. Overall, bats using hibernacula in summer can harbor Pd on their skin and in their guano, and demonstration of Pd on clothing, traps, and other equipment used at hibernacula during summertime within the WNS-affected region indicates risk for pathogen dispersal during the nonhibernal season.

Indiana, Kentucky, Ohio, Tennessee, Virginia

Sunlight, season, snowmelt, storm, and source affect E. coli populations in an artificially ponded stream

Reducing fecal indicator bacteria, such as Escherichia coli (E. coli), in streams is important for many downstream areas. E. coli concentrations within streams may be reduced by intervening ponds or wetlands through a number of physical and biological means. A section of Dunes Creek, a small coastal stream of southern Lake Michigan, was impounded and studied for 30??months from pre-through post-construction of the experimental pond. E. coli reduction became more predictable and effective with pond age. E. coli followed the hydrograph and increased several-fold during both rainfall and snowmelt events. Seasonally, the pond was more effective at reducing E. coli during summer than winter. Late summer, non-solar reduction or inactivation of E. coli in the pond was estimated at 72% and solar inactivation at 26%. E. coli DNA fingerprinting demonstrated that the winter population was genetically more homogeneous than the summer population. Detection of FRNA coliphages suggests that there was fecal contamination during heavy rain events. An understanding of how environmental factors interact with E. coli populations is important for assessing anticipated contaminant loading and the reduction of indicator bacteria in downstream reaches. ?? 2007.

Science of the Total Environment

Bacterial pathogen gene abundance and relation to recreational water quality at seven Great Lakes beaches

Quantitative assessment of bacterial pathogens, their geographic variability, and distribution in various matrices at Great Lakes beaches are limited. Quantitative PCR (qPCR) was used to test for genes from E. coli O157:H7 ( eae O157 ), shiga-toxin producing E. coli ( stx2 ), Campylobacter jejuni ( mapA ), Shigella spp. ( ipaH ), and a Salmonella enterica -specific ( SE ) DNA sequence at seven Great Lakes beaches, in algae, water, and sediment. Overall, detection frequencies were mapA > stx2 > ipaH > SE > eae O157 . Results were highly variable among beaches and matrices; some correlations with environmental conditions were observed for mapA , stx2 , and ipaH detections. Beach seasonal mean mapA abundance in water was correlated with beach seasonal mean log 10 E. coli concentration. At one beach, stx2 gene abundance was positively correlated with concurrent daily E. coli concentrations. Concentration distributions for stx2 , ipaH , and mapA within algae, sediment, and water were statistically different (Non-Detect and Data Analysis in R). Assuming 10, 50, or 100% of gene copies represented viable and presumably infective cells, a quantitative microbial risk assessment tool developed by Michigan State University indicated a moderate probability of illness for Campylobacter jejuni at the study beaches, especially where recreational water quality criteria were exceeded. Pathogen gene quantification may be useful for beach water quality management.

Environmental Science & Technology