Splenic hemangioendothelial neoplasm in a captive Northern Snakehead from the Potomac River.

OBJECTIVE: At the U.S. Geological Survey Leetown Research Laboratory in Kearneysville, West Virginia, an approximately 3-year-old, captive-held Northern Snakehead Channa argus with clinical signs of abdominal distention died and was necropsied 1 day after an examination under anesthesia. A mass discovered in the midcoelomic cavity, presumed to be deformed spleen, was comprised of large, pseudocystic structures that contained considerable volumes of opaque, straw-colored fluid. METHODS: A histopathological evaluation revealed that the tissue consisted of foci of small capillaries, nodular areas of proliferating, pleomorphic endothelial cells, and areas of necrosis within the pseudocyst wall. Positive nuclear and nonspecific immunolabeling with a vascular marker, cluster of differentiation 31, was concentrated in and around vascular spaces. RESULT: Based on these observations, the tumor has been putatively identified as a hemangioendothelial neoplasm. CONCLUSION: This would represent the first report of a vascular tumor in a Northern Snakehead and, globally, one of the few described neoplasms identified in a member of the Channidae family.

White perch health relative to urbanization and habitat degradation in Chesapeake Bay tributaries. I. Biliary neoplasms and hepatic lesions.

White perch Morone americana (Gmelin, 1789) from the Chesapeake Bay (USA) watershed have a high incidence of liver disease, including neoplasms of bile duct origin. Fish collected seasonally from spring 2019 to winter 2020 from the urban Severn River and the more rural Choptank River were evaluated for hepatic lesions. Biliary hyperplasia (64.1%), neoplasms (cholangioma and cholangiocarcinoma, 27%), and dysplasia (24.9%) were significantly higher in Severn River fish compared to Choptank River fish (52.9, 16.2, and 15.8%, respectively). Hepatocellular lesions were less common, including foci of hepatocellular alteration (FHA, 13.3%) and hepatocellular neoplasms (1%). There was also a progressive age-related increase in copper-laden granules in hepatocytes, which was a significant risk factor for FHA and could be a source of oxidative stress in the liver. Significant risk factors for biliary neoplasms included age, bile duct fibrosis, and infections by the myxozoan parasite Myxidium murchelanoi, but the prevalence and relative intensity of M. murchelanoi infections did not differ significantly between fish populations. Hepatic disease in this species appears to be chronic and may stem from an age-related accumulation of damage, possibly from parasitic infections and contaminants such as polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), and copper. Watershed development and exposures to PCBs and PAHs were generally higher for white perch in the Severn River, but similar suites of chemical contaminants were detected in the Choptank River. A broader survey of white perch within and outside Chesapeake Bay may allow determination of the extent of biliary neoplasia in this species.

White perch health relative to urbanization and habitat degradation in Chesapeake Bay tributaries. II. Hepatic and splenic macrophage aggregates.

Macrophage aggregate (MA) abundance in fish is a useful general biomarker of contaminant exposures and environmental stress. Hepatic and splenic MAs were evaluated in semi-anadromous white perch Morone americana (Gmelin, 1789) from the urbanized Severn River (S) and the more rural Choptank River (C), Chesapeake Bay. Fish were collected from different sites in the annual migratory circuit in each river that corresponded to active spawning in late winter-early spring, summer regenerating, autumn developing, and winter spawning-capable phases. An age-associated progressive increase in the total volume of MAs (MAV) was evident in the liver and spleen. Mean hepatic MAV (range in seasonal means, C: 6.4-23.1 mm3; S: 15.7-48.7 mm3) and mean splenic MAV (C: 7.3-12.6 mm3; S: 16.0-33.0 mm3) differed significantly among seasons and were significantly greater in females and in Severn River fish. Age and river were the most influential factors, suggesting that increased MAV in Severn River fish resulted from chronic exposures to higher concentrations of environmental contaminants. Hepatic MAV was directly related to the relative volume of copper granules in the liver. Less influential factors on splenic MAV included fish condition, trematode infections, and granulomas, indicating possible functional differences in MAs by organ. While organ volumes were strongly linked to gonadosomatic index (GSI) and reproductive phase, the reason for seasonal differences in MAV was less clear. Water temperature, salinity, and dissolved oxygen were not significantly related to MAV, and indicators of reproductive phase (hepatosomatic index and GSI) were significant but less important in explaining variation in MAV.

A case study: temporal trends of environmental stressors and reproductive health of smallmouth bass (Micropterus dolomieu) from a site in the Potomac River Watershed, Maryland, USA.

Decades of poor reproductive success and young-of-the-year survival, combined with adult mortality events, have led to a decline in the smallmouth bass (SMB; Micropterus dolomieu) population in sections of the Potomac River. Previous studies have identified numerous biologic and environmental stressors associated with negative effects on SMB health. To better understand the impact of these stressors, this study was conducted at the confluence of Antietam Creek and the Potomac River from 2013 to 2019 to identify temporal changes associated with SMB reproductive health. Surface water samples were collected and analyzed for over 300 organic contaminants, including pesticides, phytoestrogens, pharmaceuticals, hormones and total estrogenicity (E2Eq). Adult SMB were collected and sampled for multiple endpoints, including gene transcripts associated with reproduction (molecular), histopathology (cellular), and organosomatic indices (tissue). In males, biomarkers of estrogenic endocrine disruption, including testicular oocytes (TO) and plasma vitellogenin (Vtg) were assessed. Numerous agriculture-related contaminants or land use patterns were associated with gene transcript abundance in both male and female SMB. Positive associations between pesticides in the immediate catchment with TO severity and E2Eq with plasma Vtg in males were identified. In males, the prevalence of TO and detectable levels of plasma Vtg, liver vitellogenin transcripts (vtg) and testis vtg were high throughout the study. Peaks of complex mixtures of numerous contaminants occurred during the spring/early summer when spawning and early development occurs and to a lesser extent in fall/winter during recrudescence. Management practices to reduce exposure during these critical and sensitive periods may enhance reproductive health of these economically important sportfishes.

Development of a Multiplex Fluorescence in Situ Hybridization Assay to Identify Coinfections in Young-of-the-Year Smallmouth Bass.

Histopathological assessments of young-of-the-year (age-0) Smallmouth Bass Micropterus dolomieu in the Susquehanna River drainage identified a high prevalence of the myxozoan Myxobolus inornatus. This myxozoan infects the connective tissue of the muscle below the skin but is sometimes observed in the esophagus and buccal cavity. In some instances, shallow infections cause breaks in the skin, which could increase the chance of opportunistic bacterial infections. Several microbial pathogens, including Flavobacterium columnare, Aeromonas spp., and Largemouth Bass virus, have also been cultured from clinically diseased young of year. A multiplex fluorescence in situ hybridization (FISH) assay was developed to determine potential colocalization of M. inornatus, Flavobacterium spp., and Aeromonas spp. infections. With FISH, 75% of age-0 Smallmouth Bass exhibited M. inornatus infections, 10% had Aeromonas spp. infections, and 5% had Flavobacterium spp. infections, while 3% had coinfections with both bacterial species and M. inornatus. The results of the multiplex FISH assay revealed a low occurrence of coinfections of Flavobacterium spp. and/or Aeromonas spp. with M. inornatus in randomly sampled individuals.

Production and characterization of a mouse monoclonal antibody against smallmouth bass (Micropterus dolomieu) IgM.

A murine monoclonal antibody (mAb, IgG2a) was produced for the detection of smallmouth bass (Micropterus dolomieu) immunoglobulin (IgM). The antibody is specific for IgM heavy chain and was shown to also recognize the Ig heavy chain of largemouth bass (Micropterus salmoides) and bluegill (Lepomis macrochirus) using Western Blot analysis of plasma from 9 teleost taxa. When applied to the analysis of smallmouth bass total plasma IgM using ELISA, the mAb was found to be effective when used in an inhibition kinetic assay.

Biological and anthropogenic influences on macrophage aggregates in white perch Morone americana from Chesapeake Bay, USA.

The response of macrophage aggregates in fish to a variety of environmental stressors has been useful as a biomarker of exposure to habitat degradation. Total volume of macrophage aggregates (MAV) was estimated in the liver and spleen of white perch Morone americana from Chesapeake Bay using stereological approaches. Hepatic and splenic MAV were compared between fish populations from the rural Choptank River (n = 122) and the highly urbanized Severn River (n = 131). Hepatic and splenic MAV increased with fish age, were greater in females from the Severn River only, and were significantly greater in fish from the more polluted Severn River (higher concentrations of polycyclic aromatic hydrocarbons, organochlorine pesticides, and brominated diphenyl ethers). Water temperature and dissolved oxygen had a significant effect on organ volumes, but not on MAV. Age and river were most influential on hepatic and splenic MAV, suggesting that increased MAV in Severn River fish resulted from chronic exposures to higher concentrations of environmental contaminants and other stressors. Hemosiderin was abundant in 97% of spleens and was inversely related to fish condition and positively related to fish age and trematode infections. Minor amounts of hemosiderin were detected in 30% of livers and positively related to concentrations of benzo[a]pyrene metabolite equivalents in the bile. This study demonstrated that hepatic and splenic MAV were useful indicators in fish from the 2 tributaries with different land use characteristics and concentrations of environmental contaminants. More data are needed from additional tributaries with a wider gradient of environmental impacts to validate our results in this species.

Reproductive health and endocrine disruption in smallmouth bass (Micropterus dolomieu) from the Lake Erie drainage, Pennsylvania, USA.

Smallmouth bass Micropterus dolomieu were sampled from three sites within the Lake Erie drainage (Elk Creek, Twentymile Creek, and Misery Bay, an embayment in Presque Isle Bay). Plasma, tissues for histopathological analyses, and liver and testes preserved in RNALater® were sampled from 30 smallmouth bass (of both sexes) at each site. Liver and testes samples were analyzed for transcript abundance with Nanostring nCounter® technology. Evidence of estrogenic endocrine disruption was assessed by the presence and severity of intersex (testicular oocytes; TO) and concentrations of plasma vitellogenin in male fish. Abundance of 17 liver transcripts associated with reproductive function, endocrine activity, and contaminant detoxification pathways and 40 testes transcripts associated with male and female reproductive function, germ cell development, and steroid biosynthesis were also measured. Males with a high rate of TO (87-100%) and plasma vitellogenin were noted at all sites; however, TO severity was greatest at the site with the highest agricultural land cover. Numerous transcripts were differentially regulated among the sites and patterns of transcript abundance were used to better understand potential risk factors for estrogenic endocrine disruption. The results of this study suggest endocrine disruption is prevalent in this region and further research would benefit to identify the types of contaminants that may be associated with the observed biological effects.

Modeling estrogenic activity in streams throughout the Potomac and Chesapeake Bay watersheds.

Endocrine-disrupting compounds (EDCs), specifically estrogenic endocrine-disrupting compounds, vary in concentration and composition in surface waters under the influence of different landscape sources and landcover gradients. Estrogenic activity in surface waters may lead to adverse effects in aquatic species at both individual and population levels, often observed through the presence of intersex and vitellogenin induction in male fish. In the Chesapeake Bay Watershed, located on the mid-Atlantic coast of the USA, intersex has been observed in several sub-watersheds where previous studies have identified specific landscape sources of EDCs in tandem with observed fish health effects. Previous work in the Potomac River Watershed (PRW), the largest basin within the Chesapeake Bay Watershed, was leveraged to build random forest regression models to predict estrogenic activity at unsampled reaches in both the Potomac River and larger Chesapeake Bay Watersheds (CBW). Model outputs including important variables, partial dependence plots, and predicted values of estrogenic activity at unsampled reaches provide insight into drivers of estrogenic activity at different seasons and scales. Using the US Environmental Protection Agency effects-based threshold of 1.0 ng/L 17 ß-estradiol equivalents, catchments predicted to exceed this value were categorized as at risk for adverse effects from exposure to estrogenic compounds and evaluated relative to healthy watersheds and recreation access locations throughout the PRW. Results show immediate catchment scale models are more reliable than upstream models, and the best predictive variables differ by season and scale. A small percentage of healthy watersheds (< 13%) and public access sites were classified as at risk using the "Total" (annual) model in the CBW. This study is the first Potomac River Watershed assessment of estrogenic activity, providing a new foundation for future risk assessment and management design efforts, with additional context provided for the entire Chesapeake Bay Watershed.

High prevalence of biliary neoplasia in white perch Morone americana: potential roles of bile duct parasites and environmental contaminants.

Recent surveys of white perch Morone americana from Chesapeake Bay, USA, revealed a high prevalence of hepatic and biliary lesions, including neoplasia, and bile duct parasites. Here, we describe lesions in the liver and gallbladder and evaluate for statistical associations among lesions, parasites, and biomarkers of chemical exposure in fish from 2 tributaries of Chesapeake Bay. Fish were collected from an estuarine site in the Choptank River (n = 122, ages 3-11), a tributary with extensive agriculture within the watershed, and the Severn River (n = 131, ages 2-16), a tributary with extensive urban development. Passive integrative samplers were deployed at the fish collection site and an upstream, non-tidal site in each river for 30 d. Intrahepatic biliary lesions observed in fish from both rivers included neoplasia (23.3%), dysplasia (16.2%), hyperplasia (46.6%), cholangitis (24.9%), and dilated ducts containing plasmodia of Myxidium sp. (24.9%). Hepatocellular lesions included foci of hepatocellular alteration (FHA, 15.8%) and neoplasia in 4 Severn River fish (2.3%). Age of fish and Myxidium sp. infections were significant risk factors for proliferative and neoplastic biliary lesions, age alone was a risk factor for FHA, and Goussia bayae infections were associated with cholangitis and cholecystitis. Lesion prevalence was higher in fish from the Severn River, which contained higher concentrations of PAHs, organochlorine pesticides, and brominated diphenyl ethers. Metabolite biomarkers indicated higher PAH exposures in Severn River fish. This study suggests Myxidium sp. as a promoter of bile duct tumors, but more data are needed to evaluate the biological effects of environmental contaminants in this species.

Hyperpigmented melanistic skin lesions of smallmouth bass Micropterus dolomieu from the Chesapeake Bay watershed.

Hyperpigmented melanistic skin lesions (HPMLs) of smallmouth bass Micropterus dolomieu are observed in the Potomac and Susquehanna rivers, Chesapeake Bay watershed, USA. Routine, nonlethal population surveys were conducted at 8 sites on the mainstem Susquehanna River and 9 on the Juniata River, a tributary of the Susquehanna River, between 2012 and 2018, and the prevalence of HPMLs was documented. A total of 4078 smallmouth bass were collected from the mainstem Susquehanna River and 6478 from the Juniata River. Lesions were primarily seen in bass greater than 200 mm, and prevalence in the Susquehanna River (8%) was higher (p < 0.001) than in the Juniata River (2%). As part of ongoing fish health monitoring projects, smallmouth bass were collected at additional sites, primarily tributaries of the Susquehanna (n = 758) and Potomac (n = 545) rivers between 2013 and 2018. Prevalence in the Susquehanna River (13%) was higher (p < 0.001) than the Potomac (3%). Microscopically, HPMLs were characterized by an increased number of melanocytes in the epidermis or within the dermis and epidermis. RNAseq analyses of normal and melanistic skin identified 3 unique sequences in HPMLs. Two were unidentified and the third was a viral helicase (E1). Transcript abundance in 16 normal skin samples and 16 HPMLs showed upregulation of genes associated with melanogenesis and cell proliferation in HPMLs. The E1 transcript was detected in 12 of the 16 melanistic areas but in no samples from normal skin. Further research will be necessary to identify the putative new virus and determine its role in melanocyte proliferation.

Malignant melanoma of brown bullhead (Ameiurus nebulosus) in Lake Memphremagog, Vermont/Quebec.

In 2012, brown bullhead (Ameiurus nebulosus) with large, raised, black growths were first reported from multiple areas within the Vermont portion of Lake Memphremagog. Subsequent surveys conducted from 2014 to 2017 at two sites within the lake indicated a prevalence of 30% in adult brown bullhead 200 mm and above total length. These lesions ranged from slightly raised smooth black areas to large nodular areas on the body surface and fins and within the oral cavity. Microscopically, these lesions were determined to be malignant melanoma with invasion into surrounding hypodermis, skeletal muscle and bone as well as metastases to gill, ovary and intestine. Liver neoplasms were also observed in 8% of the bullhead collected from Lake Memphremagog in 2015. Neither skin nor liver neoplasms were noted in Ticklenaked Pond, a site used for comparison.

Mercury bioaccumulation in freshwater fishes of the Chesapeake Bay watershed.

Chemical contaminants are a threat to the Chesapeake Bay watershed, with mercury (Hg) among the most prevalent causes of impairment. Despite this, large-scale patterns of Hg concentrations, and the potential risks to fish, wildlife, and humans across the watershed, are poorly understood. We compiled fish Hg data from state monitoring programs and recent research efforts to address this knowledge gap and provide a comprehensive assessment of fish Hg concentrations in the watershed's freshwater habitats. The resulting dataset consisted of nearly 8000 total Hg (THg) concentrations from 600 locations. Across the watershed, fish THg concentrations spanned a 44-fold range, with mean concentrations varying by 2.6- and 8.8-fold among major sub-watersheds and individual 8-digit hydrological units, respectively. Although, mean THg concentrations tended to be moderate, fish frequently exceeded benchmarks for potential adverse health effects, with 45, 48, and 36% of all samples exceeding benchmarks for human, avian piscivore, and fish risk, respectively. Importantly, the percentage of fish exceeding these benchmarks was not uniform among species or locations. The variation in fish THg concentrations among species and sites highlights the roles of waterbody, landscape, and ecological processes in shaping broad patterns in Hg risk across the watershed. We outline an integrated Hg monitoring program that could identify key factors influencing Hg concentrations across the watershed and facilitate the implementation of management strategies to mitigate the risks posed by Hg.

In vitro immune function in laboratory-reared age-0 smallmouth bass (Micropterus dolomieu) relative to diet.

Smallmouth bass (Micropterus dolomieu) are used as an indicator species in environmental monitoring and assessment studies. However, laboratory-based studies for methods development and effector assessment are limited for this species. Nutrition, a known modulator of teleost physiological responses including immune function, is a critical knowledge-gap sometimes overlooked in the design of laboratory studies. We report the results from a study evaluating a commercially available artificial pelleted diet for bass and live feed (fathead minnows). Following a six-month diet-acclimation period, age-0 smallmouth bass were assessed using morphometric measures, histologic and immune-function end points using conventional methods, miniaturized cell isolation and assay methods as well as imaging flow cytometry. Fish on the two diets did not significantly differ in length, weight, or condition factor, indicating that growth was similar in the two groups. Histologic examination revealed relatively higher levels of macrophage aggregates and accumulation of ceroid/lipofuscin in the spleen as well as hepatocyte changes in the pellet-fed group. Leukocytes from the pellet-fed group exhibited significantly elevated bactericidal activity and significantly depressed mitogen response compared to fish fed live feed. Following exposure to a known inducer of inflammatory responses, bacterial lipopolysaccharide, responses including the transition of leukocytes to an apoptotic/necrotic condition differed significantly based on diet. Histologic findings were consistent with the occurrence of diet-related oxidative stress in the pellet-fed fish. Oxidative stress can be induced by multiple factors including environmental pollutants. For a diet to be useful in laboratory-based studies, it cannot elicit response that could also be induced by experimental treatments. To do so greatly complicates the detection of experimental effects. Until an artificial diet is developed for smallmouth bass that does not produce potentially confounding conditions for laboratory-based studies, use of a live feed appears to be the best option.

Seasonal development of the coccidian parasite Goussia bayae and hepatobiliary histopathology in white perch Morone americana from Chesapeake Bay.

The coccidium Goussia bayae infects the gallbladder and bile ducts of white perch Morone americana from Chesapeake Bay, USA. Seasonal changes in coccidian infections were analyzed from bile specimens of 1588 fish from the Choptank River during 2016-2018 using wet mount preparations with a Sedgwick-Rafter counting chamber. Histopathology of the gallbladder and liver was analyzed from a subset (n = 480) of these fish. Maximum parasite prevalence (100%) and intensities in the gallbladder occurred during the fish spawning season in March and April. Asynchronous coccidian development and prevalence of infections in fish increased gradually during autumn and winter, but coccidian intensity increased sharply 2-4 wk prior to the onset of fish spawning activity and decreased after spawning activity concluded. Sporulation was internal, and the gallbladder was the primary reservoir for oocysts. Two previously undescribed species of coccidia were observed in the intestine. Lesions in the gallbladder were rare and included cholecystitis and epithelial necrosis. Intrahepatic bile duct lesions were more common and included distension, cholangitis, epithelial erosion and necrosis, cholestasis, hyperplasia, and neoplasia. Cholangitis and necrosis of intrahepatic bile ducts were significantly associated with coccidial infections, while plasmodia of a myxosporean (spore morphology consistent with the genera Myxidium and Zschokella) were significantly associated with bile duct hyperplasia. Biliary neoplasia included cholangiomas (5% prevalence) and cholangiocarcinomas (1% prevalence). No association was detected between G. bayae and biliary neoplasms, but an association may exist between these lesions and the myxosporean plasmodia.

Comparisons of Stereological and Other Approaches for Quantifying Macrophage Aggregates in Piscine Spleens.

Macrophage aggregates (MAs) are focal accumulations of pigmented macrophages in the spleen and other tissues of fish. A central role of MAs is the clearance and destruction of degenerating cells and recycling of some cellular components. Macrophage aggregates also respond to chemical contaminants and infectious agents and may play a role in the adaptive immune response. Tissue damage or physiological stress can result in increased MA accumulation. As a result, MAs may be sensitive biomarkers of environmental stress in fish. Abundance of MAs in tissues has been reported in a variety of ways-most commonly as density, mean size, and relative area-but the utility of these estimates has not been compared. In this study, four different types of splenic MA abundance estimates (abundance score, density, relative area, and total volume) were compared in two fish populations (Striped Bass Morone saxatilis and White Perch M. americana) with a wide range in ages. Stereological estimates of total volume indicated an increase in MA abundance with spleen volume, which generally corresponded to fish age, and with splenic infections (mycobacteria or trematode parasites). Abundance scores were generally limited in the ability to detect changes in MA abundance by these factors, whereas density estimates were greatly influenced by changes in spleen volume. In some instances, densities declined while the total volume of MAs and spleen volume increased. Experimentally induced acute stress resulted in a decrease in spleen volume and an increase in MA density, although the total volume of MAs remained unchanged. Relative area estimates accounted for the size and number of MAs but not for changes in organ volume. Total volume is an absolute measure of MA abundance irrespective of changes in organ volume or patterns of accumulation and may provide an improved means of quantifying MAs in the spleens of fish.

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.

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.

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 were 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 ß2, while negatively correlated with androgen receptor ß 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.

Characterization of a Novel Hepadnavirus in the White Sucker (Catostomus commersonii) from the Great Lakes Region of the United States.

UNLABELLED: The white sucker Catostomus commersonii is a freshwater teleost often utilized as a resident sentinel. Here, we sequenced the full genome of a hepatitis B-like virus that infects white suckers from the Great Lakes Region of the United States. Dideoxy sequencing confirmed that the white sucker hepatitis B virus (WSHBV) has a circular genome (3,542 bp) with the prototypical codon organization of hepadnaviruses. Electron microscopy demonstrated that complete virions of approximately 40 nm were present in the plasma of infected fish. Compared to avi- and orthohepadnaviruses, sequence conservation of the core, polymerase, and surface proteins was low and ranged from 16 to 27% at the amino acid level. An X protein homologue common to the orthohepadnaviruses was not present. The WSHBV genome included an atypical, presumptively noncoding region absent in previously described hepadnaviruses. Phylogenetic analyses confirmed WSHBV as distinct from previously documented hepadnaviruses. The level of divergence in protein sequences between WSHBV and other hepadnaviruses and the identification of an HBV-like sequence in an African cichlid provide evidence that a novel genus of the family Hepadnaviridae may need to be established that includes these hepatitis B-like viruses in fishes. Viral transcription was observed in 9.5% (16 of 169) of white suckers evaluated. The prevalence of hepatic tumors in these fish was 4.9%, and only 2.4% of fish were positive for both virus and hepatic tumors. These results are not sufficient to draw inferences regarding the association of WSHBV and carcinogenesis in white sucker. IMPORTANCE: We report the first full-length genome of a hepadnavirus from fishes. Phylogenetic analysis of this genome indicates divergence from genomes of previously described hepadnaviruses from mammalian and avian hosts and supports the creation of a novel genus. The discovery of this novel virus may better our understanding of the evolutionary history of hepatitis B-like viruses of other hosts. In fishes, knowledge of this virus may provide insight regarding possible risk factors associated with hepatic neoplasia in the white sucker. This may also offer another model system for mechanistic research.