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Heather L. Walsh

Publications and source records attributed to Heather L. Walsh.

At least 37 records · Page 2Linked to original sources

Identification of Aphanomyces invadans, the cause of epizootic ulcerative syndrome, in smallmouth bass (Micropterus dolomieu) from the Cheat River, West Virginia, USA

The oomycete Aphanomyces invadans (Saprolegniales, Oomycetes), the cause of epizootic ulcerative syndrome (EUS), is an OIE (World Organization for Animal Health) reportable pathogen, capable of infecting many fish species worldwide in both freshwater and estuarine environments (Iberahim et al. 2018). Since the discovery of EUS in Japan in 1971 (Egusa and Masuda 1971), it has spread globally and caused substantial fish mortalities and economic losses (Lilley et al. 1998). Cases in the United States have included lesions initially named ulcerative mycosis of Atlantic menhaden Brevoortia tyrannus (Noga et al. 1988) along the Atlantic coast and later confirmed as A. invadans in menhaden from the Chesapeake Bay (Blazer et al. 1999). It has also been reported in several freshwater and estuarine fishes from Florida, including largemouth bass Micropterus salmoides (Sosa et al. 2007) and channel catfish, black bullhead, and bluegill from recreational ponds in Louisiana (Hawke et al. 2003).This communication reports the finding of A. invadans in smallmouth bass Micropterus dolomieu from the Cheat River, West Virginia. During fish health assessments in October 2020, smallmouth bass with grossly observable skin lesions were determined to be infected with A. invadans . We believe this is the first report of A. invadans infection in smallmouth bass and it was not observed during previous health assessments in this region.

West Virginia

Perfluoroalkyl substances in plasma of smallmouth bass from the Chesapeake Bay Watershed

Smallmouth bass Micropterus dolomieu is an economically important sportfish and within the Chesapeake Bay watershed has experienced a high prevalence of external lesions, infectious disease, mortality events, reproductive endocrine disruption and population declines. To date, no clear or consistent associations with contaminants measured in fish tissue or surface water have been found. Therefore, plasma samples from two sites in the Potomac River and two in the Susquehanna River drainage basins, differing in land-use characteristics, were utilized to determine if perfluoroalkyl substances were present. Four compounds, perfluorooctane sulphonic acid (PFOS), perfluoroundecanoic acid (PFUnA), perfluorodecanoic acid (PFDA) and perfluorododecanoic acid (PFDoA), were detected in every fish. Two additional compounds, perfluorooctane sulphonamide (PFOSA) and perfluorononanoic acid (PFNA), were less commonly detected at lower concentrations, depending on the site. Concentrations of PFOS (up to 574 ng/mL) were the highest detected and varied significantly among sites. No seasonal differences (spring versus fall) in plasma concentrations were observed. Concentrations of PFOS were not significantly different between the sexes. However, PFUnA and PFDoA concentrations were higher in males than females. Both agricultural and developed land-use appeared to be associated with exposure. Further research is needed to determine if these compounds could be affecting the health of smallmouth bass and identify sources.

New York, Pennsylvania, Maryland, Virginia

Retrospective analysis of estrogenic endocrine disruption and land-use influences in the Chesapeake Bay watershed

The Chesapeake Bay is the largest estuary in the United States and its watershed includes river drainages in six states and the District of Columbia. Sportfishing is of major economic interest, however, the rivers within the watershed provide numerous other ecological, recreational, cultural and economic benefits, as well as serving as a drinking water source for millions of people. Consequently, major fish kills and the subsequent finding of estrogenic endocrine disruption (intersex or testicular oocytes and plasma vitellogenin in male fishes) raised public and management concerns. Studies have occurred at various sites within the Bay watershed to document the extent and severity of endocrine disruption, identify risk factors and document temporal and spatial variability. Data from these focal studies, which began in 2004, were used in CART (classification and regression trees) analyses to better identify land use associations and potential management practices that influence estrogenic endocrine disruption. These analyses emphasized the importance of scale (immediate versus upstream catchment) and the complex mixtures of stressors which can contribute to surface water estrogenicity and the associated adverse effects of exposure. Both agricultural (percent cultivated, pesticide application, phytoestrogen cover crops) and developed (population density, road density, impervious surface) land cover showed positive relationships to estrogenic indicators, while percent forest and shrubs generally had a negative association. The findings can serve as a baseline for assessing ongoing restoration and management practices.

Chesapeake Bay river watersheds

Assessing native fish restoration potential in Catoctin Mountain Park

Biological conservation is a fundamental purpose of the National Park system, and Catoctin Mountain Park (CATO) supports high-quality habitat for native fishes in the headwaters of the Chesapeake Bay watershed in eastern North America. However, native Blue Ridge sculpin ( Cottus caeruleomentum ) have been extirpated in Big Hunting Creek above Cunningham Falls in CATO. Prior research indicates that infection by the fungal-like protist Dermocystidium is a likely cause for the extirpation, but elevated stream temperatures also have been observed in the study area, and it remains unknown whether thermal stress may exacerbate infections or otherwise limit habitat suitability for fishes in CATO. The purpose of this study was to quantify spatial variation in summer stream temperatures and to evaluate the effects of temperature on sculpin growth rates and susceptibility to Dermocystidium infection. We used observational and experimental methods to address these objectives. First, we deployed stream temperature gages at 10 sites throughout the study area to assess hourly and daily temperatures during the summer of 2019. Second, we conducted an in situ fish enclosure experiment at five of the temperature sites to assess fish growth and susceptibility to Dermocystidium infection over a 45-day exposure period. For this experiment we collected sculpin from a stream in CATO that supports a robust population of Blue Ridge sculpin (Owens Creek) and held them in quarantine for 50 days in the Experimental Stream Laboratory at the U.S. Geological Survey (USGS) Leetown Science Center. Pre-exposure histopathology confirmed the absence of Dermocystidium infection prior to the introduction of fish into experimental enclosures. We found that stream temperatures were warmer where sculpin have been extirpated than elsewhere in CATO where sculpin persist. However, the fish enclosure experiment revealed a positive effect of temperature on fish growth, suggesting that increased food availability and foraging rates compensated for increased metabolic demands in the warmest sites. Moreover, fish held in enclosures did not develop Dermocystidium infection. Our results therefore suggest that current environmental conditions in upper Big Hunting Creek may be suitable for Blue Ridge sculpin reintroduction, and this could ultimately lead to sportfishing opportunities by increasing the forage base for native brook trout ( Salvelinus fontinalis ).

Maryland

Development of a suite of functional immune assays and initial assessment of their utility in wild smallmouth bass health assessments

Methods were developed for measuring immune function in Micropterus dolomieu (smallmouth bass). The ultimate objective is to monitor and evaluate changes over time in immune status and disease resistance in conjunction with other characteristics of fish health and environmental stressors. To test these methods for utility in ecotoxicological studies, 192 smallmouth bass, age 2 years and older, were collected from three sites within the Susquehanna River Basin and one site in the Ohio River Basin during spring and fall 2016 and 2017. The anterior kidney was aseptically removed and homogenized for leukocyte isolation. Leukocytes were tested for bactericidal activity against two species of bacteria; respiratory burst activity when stimulated with phorbol 12-myristate 13-acetate; and mitogenesis activity when stimulated with concanavalin A, phytohemagglutinin, and lipopolysaccharide. Tissues were preserved for histopathological analyses. Two of the sites were part of a monitoring program at which surface-water samples were collected monthly (bimonthly in spring) for chemical contaminants. Significant seasonal and (or) site differences in all three immune function tests were observed. Interpretations of seasonal trends in immune function of wild fish or correlations with environmental variables and other factors are difficult to make owing to the complex nature of the immune response and the environment. Differences in immune function could potentially be related to a variety of confounding factors; therefore, additional endpoints and repeated sampling over an extended period are essential to draw conclusions on the immune status of wild fish.

Pennsylvania

A fishery after the decline: The Susquehanna River Smallmouth Bass story

The Smallmouth Bass Micropterus dolomieu fishery in the Susquehanna River basin, Pennsylvania, is one of the most socioeconomically important fisheries in the region and has recently undergone considerable changes. These changes started in 2005, when disease was documented in young-of-the-year (age-0) Smallmouth Bass. Shortly thereafter, declines in abundance of both juveniles and adults were observed. These declines in abundance coincided with disease infections in age-0, intersex in adults, and concerns regarding contaminant exposure. Natural mortality rates, particularly for age-0, increased during this period (2005–2011), and there were concerns for the overall health of this world-class fishery. However, in recent years (2012–2017), there have been decreases in both mortality rates and external observations of disease and increases in abundance across multiple size-classes. Recent changes are encouraging for the future of the Smallmouth Bass fishery in the Susquehanna River. Yet, in light of the ever changing environmental, social, and anthropogenic influences on aquatic ecosystems, there remain concerns for Smallmouth Bass health and management. Because of this, ongoing research efforts are needed to monitor population and health changes and to conduct integrative research that considers complex relationships between organisms and their environments.

Pennsylvania

The effects of tissue fixation on sequencing and transcript abundance of nucleic acids from microdissected liver samples of smallmouth bass (Micropterus dolomieu)

There is an increasing emphasis on effects-based monitoring to document responses associated with exposure to complex mixtures of chemicals, climate change, pathogens, parasites and other environmental stressors in fish populations. For decades aquatic monitoring programs have included the collection of tissues preserved for microscopic pathology. Consequently, formalin-fixed, paraffin-embedded (FFPE) tissue can be an important reservoir of nucleic acids as technologies emerge that utilize molecular endpoints. Despite the cross-linking effects of formalin, its impact on nucleic acid quality and concentration, amplification, and sequencing are not well described. While fresh-frozen tissue is optimal for working with nucleic acids, FFPE samples have been shown to be conducive for molecular studies. Laser capture microdissection (LCM) is one technology which allows for collection of specific regions or cell populations from fresh or preserved specimens with pathological alterations, pathogens, or parasites. In this study, smallmouth bass (Micropterus dolomieu) liver was preserved in three different fixatives, including 10% neutral buffered formalin (NBF), Z-Fix® (ZF), and PAXgene® (PG) for four time periods (24 hr, 48 hr, seven days, and 14 days). Controls consisted of pieces of liver preserved in RNALater® or 95% ethanol. Smallmouth bass were chosen as they are an economically important sportfish and have been utilized as indicators of exposure to endocrine disruptors and other environmental stressors. Small liver sections were cut out with laser microdissection and DNA and RNA were purified and analyzed for nucleic acid concentration and quality. Sanger sequencing and the NanoString nCounter® technology were used to assess the suitability of these samples in downstream molecular techniques. The results revealed that of the formalin fixatives, NBF samples fixed for 24 and 48 hr were superior to ZF samples for both Sanger sequencing and the Nanostring nCounter®. The non-formalin PAXgene® samples were equally successful and they showed greater stability in nucleic acid quality and concentration over longer fixation times. This study demonstrated that small quantities of preserved tissue from smallmouth bass can be utilized in downstream molecular techniques; however, future studies will need to optimize the methods presented here for different tissue types, fish species, and pathological conditions.

PLoS ONE

Assessment of microscopic pathology in fishes collected at sites impacted by wood tar in Pennsylvania

In an effort to determine whether fish populations in an area affected by wood tar waste exhibited health effects, fish were collected and analyzed with histopathology. Multiple species, including Mottled Sculpin ( Cottus bairdii ), Creek Chub ( Semotilus atromaculatus ), White Sucker ( Catostumus commersonii ), Redside Dace ( Clinostomus elongatus ), Common Shiner ( Luxilus cornutus ), and Western Blacknose Dace ( Rhinichthys obtusus ) were sampled from a reference site, Meade Run, and potentially affected streams, Kinzua Creek and Threemile Run, in northwestern Pennsylvania. A full histopathological evaluation was conducted to identify microscopic abnormalities potentially associated with wood tar exposure. The evaluation identified primarily parasites associated with tissue changes. These included microsporidian parasites in the ovaries of Common Shiner and Western Blacknose Dace; myxozoan cysts in the muscle of Common Shiner, Creek Chub, and Western Blacknose Dace; trematode cysts in the muscle of Creek Chub, Redside Dace and Common Shiner; and coccidia in spleen or pancreas of Creek Chub and Common Shiner. Microscopic abnormalities potentially associated with chemical exposure included ceroid/lipofuscin deposits in the meninges of the olfactory lobe of the brain in Common Shiner, Western Blacknose Dace, and Creek Chub, as well as bile duct proliferation and a biliary tumor in Creek Chub. Overall, the findings did not reveal significant microscopic pathology consistent with exposure to wood tar waste.

Pennsylvania

Assessment of skin and liver neoplasms in white sucker (Catostomus commersonii) collected in the Sheboygan River Area of Concern, Wisconsin, in 2017

Two hundred adult white sucker ( Catostomus commersonii ), age 3 years and older, were collected from the lower Sheboygan River Area of Concern in 2017, during the spring spawning run. Fish were euthanized, weighed, and measured, and any visible abnormalities were documented. Pieces of raised skin lesions as well as five to eight pieces of liver were removed and preserved for histopathological analyses. Skin and liver neoplasm prevalence was determined for assessment of the Fish Tumors or Other Deformities Beneficial Use Impairment. Although 45.5 percent of the suckers had raised skin lesions, the prevalence of skin neoplasms, either papilloma or squamous cell carcinoma, was 29.5 percent. This observation was similar to the prevalence (32.6 percent) of skin neoplasms in 2012; however, the percentage of squamous cell carcinoma was higher in 2017 (9.5 percent) than in 2012 (2.1 percent). The prevalence of liver neoplasms in 2017 (8.5 percent) was similar to that in 2012 (8.3 percent).

Wisconsin

Necropsy-based wild fish health assessment

Anthropogenic influences from increased nutrients and chemical contaminants, to habitat alterations and climate change, can have significant effects on fish populations. Adverse effects monitoring, utilizing biomarkers from the organismal to the molecular level, can be used to assess the cumulative effects on fishes and other organisms. Fish health has been used worldwide as an indicator of aquatic ecosystem health. The necropsy-based fish health assessment provides data on visible abnormalities and lesions, parasites, condition and organosomatic indices. These can be compared by site, season and sex, as well as temporally, to document change over time. Severity ratings can be assigned to various observations to calculate a fish health index for more quantitative assessment. A drawback of the necropsy-based assessment is that it is based on visual observations and condition factors, which are not as sensitive as tissue and subcellular biomarkers for sublethal effects. Additionally, it is rarely possible to identify causes or risk factors associated with observed abnormalities. So, for instance a raised lesion or "tumor" on the fins, lips or body surface may be a neoplasm. However, it could also be a response to a parasite, chronic inflammation or hyperplasia of normal cells in response to an irritant. Conversely, neoplasms, certain parasites, other infectious agents and many tissue changes are not visible and so may be underestimated. However, during the necropsy-based assessment, blood (plasma), tissues for histopathology (microscopic pathology), genomics and other molecular analyses, and otoliths for aging can be collected. These downstream analyses, together with geospatial analyses, habitat assessments, water quality and contaminant analyses can all be important in comprehensive ecosystem evaluations.

Journal of Visualized Experiments

Indicators of exposure to estrogenic compounds at Great Lakes Areas of Concern: Species and site comparisons

Adverse effects resulting from potential exposure of wild fishes to estrogenic endocrine disruptors was assessed at seven United States Great Lakes Areas of Concern using biomarkers ranging from organismal (gonadosomatic indices) to tissue/plasma (histology, plasma vitellogenin) and molecular (hepatic gene transcripts) levels. Biomonitoring was conducted on pelagic, top predator species, largemouth Micropterus salmoides and smallmouth M. dolomieu bass and benthic, omnivorous white sucker Catostomus commersonii. Seasonal (spring and fall) comparisons were conducted at select sites. Intersex (testicular oocytes), plasma vitellogenin and hepatic vitellogenin transcripts were commonly observed in bass species. Testicular oocyte severity was positively, although weakly, correlated with plasma vitellogenin, hepatic transcripts of vitellogenin, estrogen receptor α and estrogen receptor β2, while negatively correlated with androgen receptor β and phosphoenolpyruvate carboxykinase. No testicular oocytes were observed in white sucker, however, plasma vitellogenin and hepatic vitellogenin transcripts were commonly detected in the males. The results demonstrate the importance of utilizing multiple endpoints to assess exposure to estrogenic compounds as well as the importance of choosing sensitive species.

Great Lakes

Spatial and temporal variability of myxozoan parasite, Myxobolus inornatus, prevalence in young of the year smallmouth bass in the Susquehanna River Basin, Pennsylvania

A myxozoan parasite, Myxobolus inornatus, is one disease agent identified in young of the year (YOY) smallmouth bass in the Susquehanna River Basin, Pennsylvania. We investigated spatial and temporal variability in M. Inornatus prevalence across the Susquehanna River Basin and at several out-of-basin sites. We examined potential land use drivers of M. Inornatus prevalence including agricultural and developed land use. In 1,267 YOY smallmouth bass collected from 32 sites during 2013–2016, M. Inornatus was documented in 43.6% of samples. Among-site variability in parasite prevalence was greater than among-year variability. The effect of agricultural land use on M. Inornatus prevalence had a high probability of being positively correlated at multiple spatial scales (probability of positive effect > 0.80). The effect of developed land use on M. Inornatus prevalence had a relatively high probability of being negatively correlated at multiple spatial scales (probability of negative effect > 0.70). Our results suggest that land use practices could be related to M. Inornatus infection of smallmouth bass. Further study will be necessary to determine whether disease dynamics are a consequence of effects on the host, alterations of instream habitat mediating invertebrate host dynamics and/or survival and dispersal of the parasite infective stage.

Pennsylvania

Assessment of skin and liver neoplasms in brown bullhead (Ameiurus nebulosus) collected at the Ashtabula River Area of Concern and associated reference site, Ohio, in 2016

Brown bullhead (Ameiurus nebulosus) is a commonly used indicator species for tumor surveys at Great Lakes Areas of Concern. The “fish tumors or other deformities” is one of the beneficial use impairments at the Ashtabula River Area of Concern. In May 2016, 150 brown bullhead were collected in the lower Ashtabula River and 150 were collected in the nearby Conneaut Creek as a reference. Length, weight and external visible abnormalities were documented. Fish were euthanized, and skin lesions and liver tissue preserved for histopathological analyses. Otoliths were collected for age analyses. The percentage of bullhead with raised external lesions on lips, barbels and body surface was 34.7 percent at the Ashtabula River and 23.3 percent at Conneaut Creek. At the Ashtabula River, 26.7 percent of the bullhead collected had skin neoplasms, including papillomas, melanomas and squamous cell carcinomas, whereas at Conneaut Creek 18.6 percent had only papillomas, benign skin tumors. Liver neoplasms were observed in 7.3 percent of the bullhead from the Ashtabula River and 4.7 percent of those from Conneaut Creek. These neoplasms were observed in fish 6 years of age or older at both sites.

Ohio

Risk factors associated with mortality of age-0 Smallmouth Bass in the Susquehanna River basin, Pennsylvania

Evidence of disease and mortalities of young of the year (age‐0) Smallmouth Bass Micropterus dolomieu has occurred during the late spring and summer in many parts of the Susquehanna River watershed since 2005. To better understand contributing factors, fish collected from multiple areas throughout the watershed as well as out‐of‐basin reference populations (Allegheny and Delaware River basins; experimental ponds, Kearneysville, West Virginia) were examined grossly and histologically for abnormalities. Tissue contaminant concentrations were determined from whole‐body homogenates, and water contaminant concentrations were estimated using time‐integrated passive samplers at selected sites. Observed or isolated pathogens included bacteria, predominantly motile Aeromonas spp. and Flavobacterium columnare ; largemouth bass virus, and parasites, including trematode metacercariae, cestodes, and the myxozoan Myxobolus inornatus . Although these pathogens were found in age‐0 Smallmouth Bass from multiple sites, no one pathogen was consistently associated with mortality. Chemicals detected in tissue included polychlorinated biphenyl (PCB) congeners, organochlorine, and current‐use pesticides. Pyraclostrobin, PCB congeners 170 and 187, cis ‐chlordane and trans ‐nonachlor were detected in all Susquehanna watershed samples but rarely in samples from the reference site. The findings support the idea that there is no single cause for disease of age‐0 Smallmouth Bass; rather the cumulative effects of co‐infections and potential immunomodulation by environmental stressors during a sensitive developmental life stage may lead to mortality. Identifying the most important risk factors will be necessary for more in‐depth analyses of individual stressors and better management of the habitat and fish populations.

Pennsylvania

Pharmaceuticals, hormones, pesticides, and other bioactive contaminants in water, sediment, and tissue from Rocky Mountain National Park, 2012–2013

Pharmaceuticals, hormones, pesticides, and other bioactive contaminants (BCs) are commonly detected in surface water and bed sediment in urban and suburban areas, but these contaminants are understudied in remote locations. In Rocky Mountain National Park (RMNP), Colorado, USA, BCs may threaten the reproductive success and survival of native aquatic species, benthic communities, and pelagic food webs. In 2012–2013, 67 water, 57 sediment, 63 fish, 10 frog, and 12 quality-control samples (8 water and 4 sediment) were collected from 20 sites in RMNP. Samples were analyzed for 369 parameters including 149 pharmaceuticals, 22 hormones, 137 pesticides, and 61 other chemicals or conditions to provide a representative assessment of BC occurrence within RMNP. Results indicate that BCs were detected in water and/or sediment from both remote and more accessible locations in RMNP. The most commonly detected BCs in water were caffeine, camphor, para-cresol, and DEET; and the most commonly detected BCs in sediment were indole, 3-methyl-1H-indole, para-cresol, and 2,6-dimethyl-naphthalene. Some detected contaminants, including carbaryl, caffeine, and oxycodone, are clearly attributable to direct local human input, whereas others may be transported into the park atmospherically (e.g., atrazine) or have local natural sources (e.g., para-cresol). One or more pharmaceuticals were detected in at least 1 sample from 15 of 20 sites. Most of the 29 detected pharmaceuticals are excreted primarily in human urine, not feces. Elevated net estrogenicity was observed in 18% of water samples, and elevated vitellogenin in blood was observed in 12% of male trout, both evidence of potential endocrine disruption. Hormone concentrations in sediment tended to be greater than concentrations in water. Most BCs were observed at concentrations below those not expected to pose adverse effects to aquatic life. Results indicate that even in remote locations aquatic wildlife can be exposed to pharmaceuticals, hormones, pesticides, and other bioactive contaminants.

Colorado

Temporal evaluation of estrogenic endocrine disruption markers in smallmouth bass (Micropterus dolomieu) reveals seasonal variability in intersex

A reconnaissance project completed in 2009 identified intersex and elevated plasma vitellogenin in male smallmouth bass inhabiting the Missisquoi River, VT. In an attempt to identify the presence and seasonality of putative endocrine disrupting chemicals or other factors associated with these observations, a comprehensive reevaluation was conducted between September 2012 and June 2014. Here, we collected smallmouth bass from three physically partitioned reaches along the river to measure biomarkers of estrogenic endocrine disruption in smallmouth bass. In addition, polar organic chemical integrative samples (POCIS) were deployed to identify specific chemicals associated with biological observations. We did not observe biological differences across reaches indicating the absence of clear point source contributions to the observation of intersex. Interestingly, intersex prevalence and severity decreased in a stepwise manner over the timespan of the project. Intersex decreased from 92.8% to 28.1%. The only significant predictor of intersex prevalence was year of capture, based on logistic regression analysis. The mixed model of fish length and year-of-capture best predicted intersex severity. Intersex severity was also significantly different across late summer and early spring collections indicating seasonal changes in this metric. Plasma vitellogenin and liver vitellogenin Aa transcript abundance in males did not indicate exposure to estrogenic endocrine disrupting chemicals at any of the four sample collections. Analysis of chemicals captured by the POCIS as well as results of screening discrete water samples or POCIS extracts did not indicate the contribution of appreciable estrogenic chemicals. It is possible that unreported changes in land-use activity have ameliorated the problem, and our observations indicate recovery. Regardless, this work clearly emphasizes that single, snap shot sampling for intersex may not yield representative data given that the manifestation of this condition within a population can change dramatically over time.

Vermont

Evidence of estrogenic endocrine disruption in smallmouth and largemouth bass inhabiting Northeast U.S. National Wildlife Refuge waters: A reconnaissance study

Intersex as the manifestation of testicular oocytes (TO) in male gonochoristic fishes has been used as an indicator of estrogenic exposure. Here we evaluated largemouth bass ( Micropterus salmoides ) or smallmouth bass ( Micropterus dolomieu ) form 19 National Wildlife Refuges (NWRs) in the Northeast U.S. inhabiting waters on or near NWR lands for evidence of estrogenic endocrine disruption. Waterbodies sampled included rivers, lakes, impoundments, ponds, and reservoirs. Here we focus on evidence of endocrine disruption in male bass evidenced by gonad histopathology including intersex or abnormal plasma vitellogenin (Vtg) concentrations. During the fall seasons of 2008–2010, we collected male smallmouth bass ( n =118) from 12 sites and largemouth bass ( n =173) from 27 sites. Intersex in male smallmouth bass was observed at all sites and ranged from 60% to 100%; in male largemouth bass the range was 0–100%. Estrogenicity, as measured using a bioluminescent yeast reporter, was detected above the probable no effects concentration (0.73 ng/L) in ambient water samples from 79% of the NWR sites. Additionally, the presence of androgen receptor and glucocorticoid receptor ligands were noted as measured via novel nuclear receptor translocation assays. Mean plasma Vtg was elevated (>0.2 mg/ml) in male smallmouth bass at four sites and in male largemouth bass at one site. This is the first reconnaissance survey of this scope conducted on US National Wildlife Refuges. The baseline data collected here provide a necessary benchmark for future monitoring and justify more comprehensive NWR-specific studies.

Maine, Maryland, Massachusetts, New Hampshire, New

Monitoring of wild fish health at selected sites in the Great Lakes Basin: methods and preliminary results

During fall 2010 and spring 2011, a total of 119 brown bullhead ( Ameiurus nebulosus ), 136 white sucker ( Catostomus commersoni ), 73 smallmouth bass ( Micropterus dolomieu ), and 59 largemouth bass ( M. salmoides ) were collected from seven Great Lakes Basin Areas of Concern and one Reference Site. Comprehensive fish health assessments were conducted in order to document potential adverse affects from exposure to complex chemical mixtures. Fish were necropsied on site, blood samples obtained, pieces of liver, spleen, kidney, gill and any abnormalities placed in fixative for histopathology. Liver samples were saved for gene expression analysis and otoliths were removed for aging. A suite of fish health indicators was developed and implemented for site comparisons and to document seasonal effects and species differences in response to environmental conditions. Organism level (grossly visible lesions, condition factor), tissue level (microscopic pathology, organosomatic indices, micronuclei, and other nuclear abnormalities), plasma factors (reproductive steroid hormones, vitellogenin), and molecular (gene expression) indicators were included. This report describes the methods and preliminary results.

Great Lakes Basin