Testicular Oocytes Research Articles

Reproductive endocrine disruption and gonadal intersex induction in male Japanese medaka chronically exposed to betamethasone at environmentally relevant levels

The broad utilization of betamethasone in medical treatments may pose a significant ecotoxicological risk to aquatic organisms, yet its potential reproductive toxicity remains unclear. The present study examined the impacts of environmental exposure on male reproduction using Japanese medaka (Oryzias latipes). After 110 days of betamethasone exposure at environmentally relevant concentrations (0, 20 and 200 ng/L), LH/FSH synthesis and release in the pituitary was inhibited, and the production of sex hormones and their signaling pathways in the gonads of male medaka were greatly influenced. This synthetic glucocorticoid restrained testosterone (T) synthesis and gave rise to a significant increase in E2/T and E2/11-KT ratios. Furthermore, chronic betamethasone exposure (20 and 200 ng/L) led to the suppression of androgen receptor (AR) signaling and enhancement of estrogen receptors (ERs) signaling. An increase in hepatic vitellogenin contents was also detected, and testicular oocytes were observed in both 20 and 200 ng/L betamethasone-treated groups. It showed that 20 and 200 ng/L betamethasone could induce male feminization and even intersex, triggering abnormal spermatogenesis in medaka males. With its adverse effects on male fertility, betamethasone could potentially influence the fishery productivity and population dynamics in aquatic ecosystems.

A multi-level assessment of biological effects associated with mercury concentrations in smallmouth bass, Micropterus dolomieu

Total mercury (THg) was measured in muscle (fillet) and liver tissue of adult smallmouth bass Micropterus dolomieu collected at multiple sites in the Potomac and Susquehanna River drainages within the Chesapeake Bay watershed. Smallmouth bass in these drainages have experienced episodic mortality events, a high prevalence of skin lesions and reproductive endocrine disruption (intersex or testicular oocytes and plasma vitellogenin in males). A multi-level assessment of general and reproductive health including indicators at the organismal, organ, cellular and molecular levels was conducted on adult smallmouth bass during the spring (prespawn) season. Concentrations of THg were correlated with increased visible abnormalities, increased macrophage aggregates and tissue parasite burdens. In male bass positive correlations of THg were observed with plasma vitellogenin and hepatic transcript abundance of estrogen receptor β1 and androgen receptor α, while there was a negative association with estrogen receptors α and β2 and androgen receptors β. In female bass there was a negative correlation between THg and plasma vitellogenin as well as hepatic transcript abundance of vitellogenin, choriogenin, estrogen receptor β2 and 17β hydroxysteroid dehydrogenase. Associations of THg concentrations with various biological indicators suggest mercury may be an important environmental stressor contributing to the observed adverse effects in smallmouth bass populations.

Butachlor at environmentally relevant concentrations induces partial feminization in male Luzon wart frog Fejervarya vittigera Wiegmann, 1834

Exposure to herbicides and pesticides is linked to adverse effects on amphibian populations. Butachlor (C17H26ClNO2) is one of the common herbicides used in rice fields in the Philippines and Southeast Asia. The Luzon wart frog Fejervarya vittigera is abundant at high densities in rice fields in the Philippines. Adult F. vittigera are exposed to agriculturally relevant concentrations of butachlor for seven days under laboratory conditions to assess the effect of exposure on the histology of the testis and ovaries. Six groups: dechlorinated water (control), 0.4 mg/L, 0.8 mg/L, 1.6 mg/L, 3.2 mg/L, and 4.8 mg/L butachlor were used as test treatments, and morphology of the oocytes and seminiferous tubules and spermatogenic cells in adult F. vittigera. Partial feminization, as seen by the appearance of testicular (perinucleolar) oocytes, was observed in the testis of frogs exposed to realistic concentrations of butachlor. Overall results suggest that butachlor may interfere with normal reproductive conditions and may cause irregularities in the male reproductive tissues and the hormonal milieu.

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.

Potential health effects of contaminant mixtures from point and nonpoint sources on fish and frogs in the New Jersey Pinelands

Aquatic ecosystems convey complex contaminant mixtures from anthropogenic pollution on a global scale. Point (e.g., municipal wastewater) and nonpoint sources (e.g., stormwater runoff) are both drivers of contaminant mixtures in aquatic habitats. The objectives of this study were to identify the contaminant mixtures present in surface waters impacted by both point and nonpoint sources, to determine if aquatic biota (amphibian and fish) health effects (testicular oocytes and parasites) occurred at these sites, and to understand if differences in biological and chemical measures existed between point (on-stream) and nonpoint sources (off-stream). To accomplish this, water chemistry, fishes, and frogs were collected from 21 sites in the New Jersey Pinelands, United States. Off-stream sites consisted of 3 reference and 10 degraded wetlands. On-stream sites consisted of two reference lakes and six degraded streams/lakes (four sites above and two sites below wastewater outfalls). Surface water was collected four times at each site and analyzed for 133 organic and inorganic contaminants. One native and five non-native fish species were collected from streams/lakes and native green frogs from wetlands (ponds and stormwater basins). Limited differences in contaminant concentrations were observed in reference and degraded wetlands but for streams/lakes, results indicated that landscape alteration, (upland agricultural and developed land) was the primary driver of contaminant concentrations rather than municipal wastewater. Incidence of estrogenic endocrine disruption (intersex) was species dependent with the highest prevalence observed in largemouth bass and black crappie and the lowest prevalence observed in green frogs and tessellated darters. Parasite prevalence was site and species dependent. Prevalence of eye parasites increased with increasing concentrations of industrial, mycotoxin, and cumulative inorganic contaminants. These findings are critical to support the conservation, protection, and management of a wide range of aquatic species in the Pinelands and elsewhere as habitat loss, alteration, and fragmentation increase with increasing development.

Gonadal Development in Smallmouth Bass (Micropterus Dolomieu) Reared in the Absence and Presence of 17-α-Ethinylestradiol.

Testicular oocytes in wild adult bass (Micropterus spp.) are considered a potential indication of exposure to estrogenic compounds in municipal, agricultural, or industrial wastewater. However, our ability to interpret links between testicular oocyte occurrence in wild fish species and environmental pollutants is limited by our understanding of normal and abnormal gonadal development. We previously reported low-to-moderate testicular oocyte prevalence (7%-38%) among adult male bass collected from Minnesota waters with no known sources of estrogenic compounds. In the present study, two experiments were conducted in which smallmouth bass (Micropterus dolomieu) fry were exposed to control water or 17-α-ethinylestradiol (EE2) during gonadal differentiation, then reared in clean water for an additional period. Histological samples were evaluated at several time points during the exposure and grow-out periods, and the sequence and timing of gonadal development in the presence of estrogen were compared with that of control fish. Testicular oocytes were not observed in any control or EE2-exposed fish. Among groups exposed to 1.2 or 5.1 ng/L EE2 in Experiment 1 or 3.0 ng/L EE2 in Experiment 2, ovaries were observed in 100% of fish up to 90 days after exposure ceased, and approximately half of those ovaries had abnormal characteristics, suggesting that they likely developed in sex-reversed males. Groups exposed to 0.1, 0.4, or 1.0 ng/L in Experiment 2 developed histologically normal ovaries and testes in proportions not significantly different from 1:1. These findings suggest that, while presumably able to cause sex reversal, juvenile exposure to EE2 may not be a unique cause of testicular oocytes in wild bass, although the long-term outcomes of exposure are unknown. Environ Toxicol Chem 2022;41:1416-1428. © 2022 SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

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.

Testicular oocyte development and other toxic effects in the common hourglass tree frog exposed to three commercial insecticide formulations

The Ceylon Journal of Science is published by the University of Peradeniya, Sri Lanka. Full text available. The journal also has its own website. The Ceylon Journal of Science is a continuation of the Ceylon Journal of Science (Biological Sciences) which is no longer being published as a separate journal. The history of the journal can be found here.From May 2020, Ceylon Journal of Science is indexed in DOAJ.

Health status of fathead minnow (Pimephales promelas) populations in a municipal wastewater effluent-dominated stream in the Canadian prairies, Wascana Creek, Saskatchewan.

Their unique hydrological and climatic conditions render surface water systems in the southern Canadian Prairies at an elevated risk from exposure to contaminants released from municipal wastewater effluents (MWWEs). The aim of this study was to characterize the potential health effects and their underlying molecular mechanisms in populations of fathead minnow (Pimephales promelas; FHM) in Wascana Creek, an effluent dominated stream in Southern Saskatchewan, Canada. Studies were conducted during the spawning season in 2014 and 2015 to assess responses in terms of overall health, reproductive functions, plasma sex steroid hormone levels, and expression of selected genes along the hypothalamus-pituitary-gonadal axis. FHM downstream of the effluent fallout had lower gonadosomatic indices and significantly greater hepatosomatic indices compared to upstream populations. In both male and female FHMs, significantly greater occurrence and severity of gonadal degradation and delayed maturation were observed in downstream fish compared to upstream fish. Downstream males also displayed lower scores of secondary sexual characteristics and a decreasing trend in plasma 11-ketotestosterone levels. Interestingly, no indications of exposure to estrogenic compounds, such as occurrence of testicular oocytes were observed, which was in accordance with the lack of presence of key biomarkers of estrogenic exposure, such as induction of vitellogenin. In general, expression of the majority of transcripts measured in FHMs downstream of the effluent fallout was significantly downregulated, which supports observations of the general deterioration of the health and reproductive status of these fish. Chemical analysis indicated that 10 pharmaceuticals and personal care products (PPCPs) were present at the downstream site, some at sufficiently great concentrations that may present a risk to aquatic organisms. With continuous exposure to a diverse number of stressors including high nutrient and ammonia levels, the presence of a variety of PPCPs and other contaminants, Wascana Creek should be considered as an ecosystem at risk.

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.

Gonad metabolomics and blood biochemical analysis reveal differences associated with testicular oocytes in wild largemouth bass (Micropterus salmoides)

Adverse reproductive effects associated with gonadal intersex among freshwater fish could hold considerable implications for population sustainability. Presence of testicular oocytes (TO) is the most common form of intersex and is widespread among centrarchids (sunfishes) of North America and other freshwater teleosts. Placing TO within the toxicological context of adverse outcome pathways (AOPs) to assess ecological risk is a priority for ecotoxicologists due to the association of TO with harmful chemical exposure and adverse reproductive effects in some cases. However, key event relationships between EDC exposure, incidence of TO, and apical outcomes have yet to be fully elucidated - in part due to a lack of knowledge of relationships between intersex gonad physiology and fish health. Understanding the physiological status of intersex fish is critical to assess ecological risk, understand mechanisms of induction, and to establish biomarkers of intersex in fish. In the present study, features of gonad metabolite profiles associated with TO in largemouth bass (LMB, Micropterus salmoides) from an impoundment in Georgia (USA) were determined using GC-MS-based metabolomics. Clinical blood biochemical screens were used to evaluate markers of fish health associated with TO. Results suggest that physiological changes in energy expenditure as well as relatively 'feminized' gonad lipid and protein metabolism may be related to the occurrence of TO in male LMB, and highlight the need to understand relationships between intersex and physical stressors such as elevated temperature and hypoxia. These results provide novel insight to AOPs associated with TO and identify candidate analytes for biomarker discovery.

SAT-732 Historical Occurrence of Intersex in Largemouth Bass (Micropterus Salmoides) and Smallmouth Bass (Micropterus Dolomieu)

The widespread occurrence of gonadal intersex (presence of testicular oocytes) has been recently reported in populations of Largemouth Bass (Micropterus salmoides) and Smallmouth Bass Micropterus dolomieu) from populations across North America. To evaluate the historical occurrence of intersex in bass species, gonads were examined visually and histologically from Largemouth Bass and Smallmouth Bass from museum specimens collected from 16 States. These fishes were collected between 1823-1965, before the widespread discharge of estrogenic chemicals to surface waters via wastewater effluents. All females examined had normal ovaries. Intersex gonads were identified in 17.8 % of male Largemouth Bass, and in 14.2% of male Smallmouth Bass. The intensity of testicular oocytes in some males was as great as observed since 2000. These results indicate that the occurrence of this form of intersex in these bass species is not a recent phenomenon.

Reconnaissance of Surface Water Estrogenicity and the Prevalence of Intersex in Smallmouth Bass (Micropterus Dolomieu) Inhabiting New Jersey

The observation of testicular oocytes in male fishes has been utilized as a biomarker of estrogenic endocrine disruption. A reconnaissance project led in the Northeastern United States (US) during the period of 2008–2010 identified a high prevalence of intersex smallmouth bass on or near US Fish & Wildlife Service National Wildlife Refuges that included the observation of 100% prevalence in smallmouth bass males collected from the Wallkill River, NJ, USA. To better assess the prevalence of intersex smallmouth bass across the state of New Jersey, a tiered reconnaissance approach was initiated during the fall of 2016. Surface water samples were collected from 101 (85 river, 16 lake/reservoir) sites across the state at base-flow conditions for estrogenicity bioassay screening. Detectable estrogenicity was observed at 90% of the sites and 64% were above the US Environmental Protection Agency trigger level of 1 ng/L. Median surface water estrogenicity was 1.8 ng/L and a maximum of 6.9 ng/L E2EqBLYES was observed. Adult smallmouth bass were collected from nine sites, pre-spawn during the spring of 2017. Intersex was identified in fish at all sites, and the composite intersex prevalence was 93.8%. Prevalence across sites ranged from 70.6% to 100%. In addition to intersex, there was detectable plasma vitellogenin in males at all sites. Total estrogenicity in surface water was determined at these fish collection sites, and notable change over time was observed. Correlation analysis indicated significant positive correlations between land use (altered land; urban + agriculture) and surface water estrogenicity. There were no clear associations between land use and organismal metrics of estrogenic endocrine disruption (intersex or vitellogenin). This work establishes a baseline prevalence of intersex in male smallmouth bass in the state of New Jersey at a limited number of locations and identifies a number of waterbodies with estrogenic activity above an effects-based threshold.

Histopathologic Prevalence and Severity of Testicular Oocytes in Smallmouth Bass from Two Archival Collections.

During recent decades, survey studies have documented the widespread presence of oocytes in the testes of male Smallmouth Bass Micropterus dolomieu collected from surface waters throughout the United States. There are few published reports of testicular oocytes (TO) in Smallmouth Bass before the 1990s, so it is difficult to know how long this has been occurring. Consequently, this study was conducted to evaluate the prevalence and severity of TO occurrence in whole fish specimens from two archival collections-the Smithsonian Institution's National Museum of Natural History in Suitland, Maryland, and Cornell University's Museum of Vertebrates in Ithaca, New York. Gonads were excised from 167 preserved male Smallmouth Bass that were originally collected between 1875 and 2004, and routine histologic sections were prepared and examined. The severity of TO was determined using a semiquantitative scoring system. Overall, 52.1% of male Smallmouth Bass were found to have TO. Affected fish had been collected in 11 of the 18 represented states, and TO were found in specimens harvested during decades as early as the 1880s and 1900s. Unfortunately, the small number of samples acquired at the earliest time periods precluded analyses of prevalence and severity trends over time. The results of this study demonstrated that the phenomenon of TO in male Smallmouth Bass is at least a century old and confirmed the widespread nature of this finding throughout the species' historic range. Further research efforts should focus on determining the baseline prevalence of TO in laboratory-reared male Smallmouth Bass that have not been exposed to endocrine active substances or the effects of experimental estrogen exposure on such fish.

Survey of Intersex (Testicular Oocytes) in Black Crappie Collected from the Illinois River Waterway

Abstract Researchers have documented intersex (testicular oocytes) in male fishes in many species worldwide and prevalence of the condition is strongly related to exposure to endocrine-disrupting compounds in aquatic ecosystems. There is no available knowledge about the incidence of intersex in Black Crappie Pomoxis nigromaculatus, an important North American sport fish. The objective of this survey was to investigate the prevalence and distribution of the intersex condition in Black Crappie collected from portions of the Illinois River waterway. We collected and examined 83 Black Crappie. We observed testicular oocytes in 22 individuals (27% intersex), representing the first documentation of intersex condition in wild Black Crappie. The number of oocytes that we observed in intersex Black Crappie ranged from 1 to 67. We documented intersex in individuals collected from all habitats, but there were no significant differences in the prevalence of the condition (Kruskal–Wallis P = 0.56) or the number of oocytes present (P = 0.25) in intersex males from the different habitats. Permutational multivariate analysis of variance indicated that morphometric parameters varied significantly among locations (P = 0.0001), but did not vary significantly between intersex and normal males. We have little information about the current distribution of endocrine-disrupting compounds in the specific regions where we collected Black Crappie for our study. However, we believe our study represents a valuable effort to detect the condition in an important sport fish and lays a foundation for future research.

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 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.

Quantifying population-level effects of water temperature, flow velocity and chemical-induced reproduction depression: A simulation study with smallmouth bass

Evaluating stochastic abiotic factors and their combined effects on fish and wildlife populations have been challenging in environmental sciences. Contributing to this challenge is the paucity of data describing how observations made on individuals related to exposure to chemical compounds ultimately effect population vital rates, and how this exposure interacts with other abiotic drivers. Using three smallmouth bass populations in Pennsylvania as a case study, we explored both single-factor and combined effects of water temperature, flow velocity and chemical exposure on populations through a model simulation. Although there are many pathways through which chemicals can affect population vital rates, we focused on one where chemical exposure leads to reduced reproduction. Among the three populations considered, two (the Juniata and Susquehanna populations) have experienced adverse health, including the potential adverse effects of environmental stress and chemical contamination that may cause disease and mortality of young-of-year (YOY), various skin lesions and a high prevalence of intersex or testicular oocytes in adults. The third population (The Alleghany population) has not encountered mortality events of YOY and intersex prevalence is much lower. The simulation involved projecting populations using a length-based model under different environmental conditions. In the simulations, abiotic factors influenced population dynamics through their impacts on growth, survival and recruitment. Response to the same environmental stress varied by population and life-stage of the species. Factors affecting young adult and adult life-stages had great influence on proportional stock density (PSD) and the probability of having PSD within the suggested range (PSD probability). Increases in water temperature had a negative effect and dominant role in the combined effect on population size structure (e.g., PSD and PSD probability) – increases in flow velocity during the spring season also had a negative effect on abundance. Populations with high recruitment rates sustained relatively large population size, even under high water temperature and/or high flow velocity, which suggests that factors and management strategies that benefit recruitment (such as reduced chemical contaminants) may compensate for the negative effects of warming water temperatures and high spring flow velocity on population size.

Hermaphroditism induction of Sub-Lethal Dose of Atrazine and Atrazine-Nitrate on the Egyptian Toad, <i>Sclerophrys Regularis</i>

In this study, atrazine has been shown to act as a potent endocrine disruptor in amphibians either alone or in combination with nitrate under stable laboratory conditions, causing altered gonadal development at the sub-lethal concentrations. A control group and three treatments were tested; atrazine, nitrate, atrazine- nitrate treatments with doses of 300 μg/L, 200 mg/L and their combination, respectively. Atrazine exhibited increase the presence of testicular oocytes TOs in most treated specimens; furthermore, the sizes of seminiferous tubules were decreased compared to control group. Conversely no intersex individuals were detected in the nitrate treatment alone. The combined effects of nitrate and atrazine were not additive or synergistic but were similar to the effects of atrazine in raising the percentage of intersex but with increasing the size of testicular oocyte compared to atrazine alone. On the other hand, the control group in the present study did not contain the rudimentary testicular oocytes and showed normal structures. These gonadal abnormalities may reduce reproductive success and could be a major factor behind amphibian declines. Also, these negative impacts may be a bioindicator alarming the ecosystem disrupting caused by the uncontrolled apply of these chemicals in agriculture.

The occurrence of intersex fishes in Boulder Creek, Colorado, is a recent phenomenon

Gonads were examined visually and histologically from white sucker (Catostomus commersoni) and fathead minnows (Pimephales promelas) isolated from museum specimens collected from Boulder Creek, Colorado. These fishes were collected between 42 and 102 years ago before addition of large quantities of estrogenic chemicals via wastewater effluent was reported to disrupt reproductive structures and functions in white suckers living in Boulder Creek downstream of the wastewater treatment facility (WWTF) and in test exposures of fathead minnows to wastewater effluent at the WWTF. No evidence of abnormal external gonad appearance or histology (e.g., testicular oocytes, mixed gonadal tissue) were observed in male or female museum specimens of either species supporting the conclusion that observations of reproductive abnormalities, feminization, demasculinization, and altered sex ratios are recent phenomena.

Testicular oocytes in smallmouth bass in northeastern Minnesota in relation to varying levels of human activity.

Testicular oocytes (TOs) have been found in black bass (Micropterus spp.) from many locations in North America. The presence of TOs is often assumed to imply exposure to estrogenic endocrine disrupting compounds (EDCs); however, a definitive causal relationship has yet to be established, and TO prevalence is not consistently low in fish from areas lacking evident EDC sources. This might indicate any of a number of situations: 1) unknown or unidentified EDCs or EDC sources, 2) induction of TOs by other stressors, or 3) testicular oocytes occurring spontaneously during normal development. In the present study, we analyzed TO occurrence in smallmouth bass (Micropterus dolomieu) from 8 populations in northeastern Minnesota watersheds with differing degrees of human development and, hence, presumed likelihood of exposure to anthropogenic chemicals. Three watersheds were categorized as moderately developed, based on the presence of municipal wastewater discharges and higher human population density (4-81 per km2 ), and 5 watersheds were minimally developed, with very low human population density (0-1 per km2 ) and minimal built environment. Testicular tissues from mature fish were evaluated using a semiquantitative method that estimated TO density, normalized by cross-sectional area. Testicular oocyte prevalence and density among populations from moderately developed watersheds was higher than in populations from minimally developed watersheds. However, TO prevalence was unexpectedly high and variable (7-43%) in some populations from minimally developed watersheds, and only weak evidence was found for a relationship between TO density and watershed development, suggesting alternative or more complex explanations for TO presence in smallmouth bass from this region. Environ Toxicol Chem 2017;36:3424-3435. © 2017 SETAC.