Gonadal abnormalities in frogs (\u3ci\u3eLithobates\u3c/i\u3e spp.) collected from managed wetlands in an agricultural region of Nebraska, USA

Nebraska’s Rainwater Basin (RWB) provides important wetland habitat for North American migratory birds. Concern exists that pesticide and nutrient runoff from surrounding row-crops enters wetlands degrading water quality and adversely affecting birds and wildlife. Frogs may be especially vulnerable. Plains leopard (Lithobates blairi) metamorphs from RWB wetlands with varying concentrations of pesticides were evaluated for a suite of biomarkers of exposure to endocrine active chemicals. Froglets had ovarian dysgenesis, high rates of testicular oocytes, and female-biased sex ratios however, there was no clear statistical association between pesticide concentrations and biomarkers. Data interpretation was hindered because timing and duration of exposures were unknown and due to an incomplete understanding of L. blairi sexual development. Emphasis is on describing the complex developmental biology of closely-related leopard frogs, how this understanding can explain RWB L. blairi anomalies, and the need for sampling at the appropriate life stage

Widespread occurrence of intersex in black basses (\u3ci\u3eMicropterus\u3c/i\u3e spp.) from U.S. rivers, 1995–2004

Intersex occurrence in freshwater fishes was evaluated for nine river basins in the United States. Testicular oocytes (predominantly male testes containing female germ cells) were the most pervasive form of intersex observed, even though similar numbers of male (n = 1477) and female (n = 1633) fish were examined. Intersex was found in 3% of the fish collected. The intersex condition was observed in four of the 16 species examined (25%) and in fish from 34 of 111 sites (31%). Intersex was not found in multiple species from the same site but was most prevalent in largemouth bass (Micropterus salmoides; 18% of males) and smallmouth bass (M. dolomieu; 33% of males). The percentage of intersex fish per site was 8–91% for largemouth bass and 14–73% for smallmouth bass. The incidence of intersex was greatest in the southeastern United States, with intersex largemouth bass present at all sites in the Apalachicola, Savannah, and Pee Dee River Basins. Total mercury, trans-nonachlor, p,p\u27-DDE, p,p\u27-DDD, and total PCBs were the most commonly detected chemical contaminants at all sites, regardless of whether intersex was observed. Although the genotype of the intersex fish was not determined, the microscopic appearance of the gonads, the presence of mature sperm, and the concentrations of sex steroid hormones and vitellogenin indicate the intersex bass were males. Fewre productive endpoints differed significantly among male and intersex bass; plasma vitellogenin concentration in males was not a good indicator of intersex presence. Hierarchical linkages of the intersex condition to reproductive function will require a more quantitative measure of intersex (e.g. severity index) rather than presence or absence of the condition. The baseline incidence of intersex gonadal tissue in black basses and other freshwater fishes is unknown, but intersex prevalence may be related to collection season, age, and endocrine active compounds in the environment. Intersex was not found in largemouth bass older than five years and was most common in 1–3-year-old male largemouth bass. The cause(s) of intersex in these species is also unknown, and it remains to be determined whether the intersex we observed in largemouth and smallmouth bass developed during sex differentiation in early life stages, during exposure to environmental factors during adult life stages, or both

In ovo exposure to o,p -DDE affects sexual development but not sexual differentiation in Japanese medaka (Oryzias latipes).

Despite being banned in many countries, dichlorodiphenyltrichloroethane (DDT) and its metabolites dichlorodiphenyldichloroethylene (DDE) and dichlorodiphenyldichloroethane (DDD) continue to be found in fish tissues at concentrations of concern. Like o,p -DDT, o,p -DDE is estrogenic and is believed to exert its effects through binding to the estrogen receptor. The limited toxicologic data for o,p -DDE suggest that it decreases fecundity and fertility of fishes. We conducted an egg injection study using the d-rR strain of medaka and environmentally relevant concentrations of o,p -DDE to examine its effects on sexual differentiation and development. The gonads of exposed fish showed no evidence of sex reversal or intersex. However, other gonad abnormalities occurred in exposed individuals. Females exhibited few vitellogenic oocytes and increased atresia. Male testes appeared morphologically normal but were very small. Gonadosomatic index values for both sexes were lower for exposed fish. Our observations of abnormal female and very small male gonads after in ovo o,p -DDE exposure may be indicative of effects on early endocrine processes important for normal ovarian and testicular development

Gonadal abnormalities in frogs (\u3ci\u3eLithobates\u3c/i\u3e spp.) collected from managed wetlands in an agricultural region of Nebraska, USA

Nebraska’s Rainwater Basin (RWB) provides important wetland habitat for North American migratory birds. Concern exists that pesticide and nutrient runoff from surrounding row-crops enters wetlands degrading water quality and adversely affecting birds and wildlife. Frogs may be especially vulnerable. Plains leopard (Lithobates blairi) metamorphs from RWB wetlands with varying concentrations of pesticides were evaluated for a suite of biomarkers of exposure to endocrine active chemicals. Froglets had ovarian dysgenesis, high rates of testicular oocytes, and female-biased sex ratios however, there was no clear statistical association between pesticide concentrations and biomarkers. Data interpretation was hindered because timing and duration of exposures were unknown and due to an incomplete understanding of L. blairi sexual development. Emphasis is on describing the complex developmental biology of closely-related leopard frogs, how this understanding can explain RWB L. blairi anomalies, and the need for sampling at the appropriate life stage

Bioaccumulation of Stentorin, the Probable Causative Agent for Discolored (“Purple”) Eggs and Ovaries in Blue Catfish (\u3ci\u3eIctalurus furcatus\u3c/i\u3e) from Eufaula Lake, Oklahoma, USA

Observations of reddish to “purple” discolored eggs in the ovaries of adult female blue catfish (Ictalurus furcatus) from the northern arm of Eufaula Lake, a eutrophic multiuse impoundment in east-central Oklahoma, were first reported in 2006. Blue catfish eggs are normally cream to light yellow. Reports peaked in 2007−2008 and declined through 2009−2010; purple eggs have not been reported between 2010 and 2014. In the laboratory, all tissues and fluids of affected fish were strongly orange-red fluorescent under UV illumination, with the fluorescence most apparent in the lipid-rich ovaries and eggs. The causative agent was isolated chromatographically and confirmed by mass spectrometry as stentorin (1,3,4,6,8,10,11,13- octahydroxy-2,5-diisopropyl-phenanthro[1,10,9,8,o,p,q,r,a]- perylene-7,14-dione), the fluorescent, lipophilic pigment associated with the photoreceptor protein of the ciliated protozoan Stentor coeruleus (Heterotrichea; Stentoridae). Larval medaka (Orizias latipes) readily consumed S. coeruleus in the laboratory and were observed to fluoresce in the same manner as the affected blue catfish. Potential deleterious effects of stentorin bioaccumulation remain to be determined, as do the geographic extent and the identities of other fluorescent compounds isolated from catfish eggs and ovaries

Evaluating spawning migration patterns and predicting spawning success of shovelnose sturgeon in the Lower Missouri River

Approaches using telemetry, precise reproductive assessments, and surgically implanted data storage tags (DSTs) were used in combination with novel applications of analytical techniques for fish movement studies to describe patterns in migratory behavior and predict spawning success of gravid shovelnose sturgeon. From 2004 to 2007, over 300 gravid female shovelnose sturgeon (Scaphirhynchus platorynchus) from the Lower Missouri River, that were expected to spawn in the year they were collected, were surgically implanted with transmitters and archival DSTs. Functional cluster modeling of telemetry data from the spawning season suggested two common migration patterns of gravid female shovelnose sturgeon. Fish implanted from 958 to 1181 river kilometer (rkm) from the mouth of the Missouri River (or northern portion of the Lower Missouri River within 354 rkm of the lowest Missouri River dam at rkm 1305) had one migration pattern. Of fish implanted from 209 to 402 rkm from the mouth of the Missouri River (or southern portion of the Lower Missouri River), half demonstrated a movement pattern similar to the northern fish while the other half demonstrated a migration pattern that covered more of the river. There was no apparent difference in migration patterns between successful and unsuccessful spawners. Multiple hypotheses exist to explain differences in migratory patterns among fish from different river reaches. Additional work is required to determine if observed differences are due to multiple adapted strategies, environmental alteration, and ⁄ or initial tagging date. Hierarchical Bayesian modeling of DST data indicated that variation in depth usage patterns was consistently different between successful and unsuccessful spawners, as indicated by differences in likelihood of switching between high and low variability states. Analyses of DST data, and data collected at capture, were sufficient to predict 8 of 10 non-spawners ⁄ incomplete spawners and all 30 spawners in the absence of telemetry location data. Together, the results of these two separate analyses suggest that caution is necessary in extrapolating spawning success from broad-scale movement data alone. More direct measures of spawning success may be necessary to precisely determine spawning success and to evaluate the effects of management actions

Bioaccumulation of Stentorin, the Probable Causative Agent for Discolored (“Purple”) Eggs and Ovaries in Blue Catfish (<i>Ictalurus furcatus</i>) from Eufaula Lake, Oklahoma, USA

Observations of reddish to “purple” discolored eggs in the ovaries of adult female blue catfish (Ictalurus furcatus) from the northern arm of Eufaula Lake, a eutrophic multiuse impoundment in east-central Oklahoma, were first reported in 2006. Blue catfish eggs are normally cream to light yellow. Reports peaked in 2007–2008 and declined through 2009–2010; purple eggs have not been reported between 2010 and 2014. In the laboratory, all tissues and fluids of affected fish were strongly orange-red fluorescent under UV illumination, with the fluorescence most apparent in the lipid-rich ovaries and eggs. The causative agent was isolated chromatographically and confirmed by mass spectrometry as stentorin (1,3,4,­6,8,10,­11,13-octahydroxy-2,5-diisopropyl-phenanthro­[1,10,9,­8,o,p,­q,r,a]­perylene-7,14-dione), the fluorescent, lipophilic pigment associated with the photoreceptor protein of the ciliated protozoan Stentor coeruleus (Heterotrichea; Stentoridae). Larval medaka (Orizias latipes) readily consumed S. coeruleus in the laboratory and were observed to fluoresce in the same manner as the affected blue catfish. Potential deleterious effects of stentorin bioaccumulation remain to be determined, as do the geographic extent and the identities of other fluorescent compounds isolated from catfish eggs and ovaries

A HEALTH RISK EVALUATION FOR PALLID STURGEON (\u3ci\u3eSCAPHIRHYNCHUS ALBUS\u3c/i\u3e) IN THE LOWER PLATTE RIVER USING SHOVELNOSE STURGEON (\u3ci\u3eSCAPHIRHYNCHUS PLATORYNCHUS\u3c/i\u3e) AS A SURROGATE

Most sturgeon species worldwide have been in steep decline since the 1900s. This research evaluated shovelnose sturgeon health, reproduction, and exposure to environmental contamination in the lower Platte River. Shovelnose sturgeon served as a surrogate species for the endangered pallid sturgeon and their health was assessed by incorporating measurements of general health with hepatic, immune, and reproductive system biomarkers. Environmental contaminants were measured in water, potential pallid sturgeon food items (cyprinid minnows), and shovelnose sturgeon digesta, liver, and blood plasma. Contaminants detected in shovelnose sturgeon at concentrations of concern included PCBs, selenium, and atrazine. Total PCBs in carcasses (n = 8) averaged 0.32 μg/g ww. Selenium averaged 4.8 μg/g dw in carcasses (n = 30) and 80 percent of individuals sampled were within the 4 to 6 μg/g threshold range for reproductive impairment in sensitive fish species. Pallid sturgeon food items had significantly (p \u3c 0.05) greater concentrations of Hg, Se, and Zn than shovelnose sturgeon digesta. Atrazine was detected in all blood plasma samples analyzed (n = 50) at concentrations from 0.24 to 28 μg/L, but was not detected in liver (n = 19). Although the effects of atrazine exposure to shovelnose sturgeon are unknown, results of this study and previous work by others indicate that it may be disrupting steroidogeneisis. Gross observations and condition indices seem to indicate that shovelnose sturgeon from the lower Platte River are healthy; however, reproductive biomarkers and histological examination of gonads indicate potential reproductive impairment as indicated by ovicular atresia, abnormal estrogen to testosterone ratios, and high concentrations of vitellogenin in males. Pallid sturgeon may be especially at risk to contaminants in the lower Platte River that bioaccumulate and cause reproductive impairment because they have a more piscivourus diet, greater maximum life-span, and a longer reproductive cycle than shovelnose sturgeon. Strategies to reduce shovelnose sturgeon and pallid sturgeon exposure to environmental contaminants in the lower Platte River are presented