Incidence Of Intersex Research Articles

First account of a transient intersex in spotted scat, Scatophagus argus: a marine gonochoristic fish.

This study presents the first incidence of intersex associated with testis-ova in spotted scat (Scatophagus argus) reared in a controlled environment. The testis-ova is associated with the abnormal occurrence of primary oocytes (POs) in some male testis and is referred to as ectopic primary oocytes (Ecto-PO), whiles individuals with Ecto-PO are called "Ecto-PO gonad/individuals." We investigated gonads of 129 male spotted scat aged 4-12 and 18 months after hatch (mah) by histological studies for the presence of female sexual characteristics. A total of 20 out of 88 gonads representing 22.7% of male fish aged 6-12, or 15.5% of all male fish sampled, were found to have visible Ecto-PO. At least, the Ecto-PO had an average of 7 oocytes per gonadal section, indicating high severity. The Ecto-PO appears after sex differentiation and degenerates during sexual maturation. The Ecto-PO did not significantly influence the expression pattern of male and female sex-related genes performed using qPCR. Immunofluorescence of 42sp50 specifically stained the Ecto-PO without influence from the surrounding testicular tissues. In addition, temperature did not correlate with the proliferation of the Ecto-PO, but rather gonad developmental strategy. The results show that the naturally occurring Ecto-PO might not be detrimental to testis development and could be considered a frequent-high-level incidence of natural aberration. This study highlights the intricacy of fish sex differentiation and provides a new research chapter to ascertain the mystery behind the occurrence of Ecto-PO.

The Occurrence of Intersex in Different Populations of the Marine Amphipod Echinogammarus marinus in North-West Brittany - A Longterm-Study.

In the past two decades, an increasing body of studies has been published on the intersex phenomenon in separate-sexed crustaceans from marine and freshwater ecosystems. Various causes are being considered that could have an influence on the occurrence of intersex. Besides genetic factors, environmental conditions such as photoperiodicity, temperature, salinity and parasitism, but also environmental pollution with endocrine disrupting chemicals (EDCs) are discussed. As part of a long-term monitoring (2012 – 2020) in north-west Brittany, we recorded the occurrence of intersex in the marine amphipod Echinogammarus marinus. We quantified the intersex incidence at marine and estuarine sites and analyzed the incidence in relation to the endocrine potential of the sediments. Intersex occurred with mean frequencies between 0.87% and 12%. It was striking that the incidence of intersex increased with increasing distance from the sea. Since the highest incidence was observed at the range boundary of this stenohaline species, we assume that intersex is triggered by endocrine potential and increasing stress due to increasing freshwater content − and thus an interplay of different environmental factors.

First report of intersex in Cobia, Rachycentron canadum reared in Gulf of Mannar, India—A case study

The present study reports the first incidence of intersex in Cobia, Rachycentron canadum maintained at sea cages in the Gulf of Mannar, India. During the 10 year study period (2010–2019), at total of 815 broodstock (386 wild captured and 429 captive raised) were maintained under captivity in sea cages in the Gulf of Mannar, India for the purpose of seed production. Of which, 318 male and 252 female broods’ gonads were examined post-mortem following a natural death. Only one male fish was found with a visible gonadal anomaly. Microscopic analysis of gonad revealed the presence of both the characteristic female and male tissues in testes of a single specimen (testis-ova). Only the abnormal nodular portion contained primary oocytes, whereas the central testicular tissue contained spermatocytes and spermatids in the seminiferous lobules. This is the first reported intersex of cobia from the Gulf of Mannar, India. It could be interpreted as low-level rare incidence of natural aberration.

Effects of early life stage exposure of largemouth bass to atrazine or a model estrogen (17α-ethinylestradiol)

Endocrine disrupting contaminants are of continuing concern for potentially contributing to reproductive dysfunction in largemouth and smallmouth bass in the Chesapeake Bay watershed (CBW) and elsewhere. Exposures to atrazine (ATR) have been hypothesized to have estrogenic effects on vertebrate endocrine systems. The incidence of intersex in male smallmouth bass from some regions of CBW has been correlated with ATR concentrations in water. Fish early life stages may be particularly vulnerable to ATR exposure in agricultural areas, as a spring influx of pesticides coincides with spawning and early development. Our objectives were to investigate the effects of early life stage exposure to ATR or the model estrogen 17α-ethinylestradiol (EE2) on sexual differentiation and gene expression in gonad tissue. We exposed newly hatched largemouth bass (LMB, Micropterus salmoides) from 7 to 80 days post-spawn to nominal concentrations of 1, 10, or 100 µg ATR/L or 1 or 10 ng EE2/L and monitored histological development and transcriptomic changes in gonad tissue. We observed a nearly 100% female sex ratio in LMB exposed to EE2 at 10 ng/L, presumably due to sex reversal of males. Many gonad genes were differentially expressed between sexes. Multidimensional scaling revealed clustering by gene expression of the 1 ng EE2/L and 100 µg ATR/L-treated male fish. Some pathways responsive to EE2 exposure were not sex-specific. We observed differential expression in male gonad in LMB exposed to EE2 at 1 ng/L of several genes involved in reproductive development and function, including star, cyp11a2, ddx4 (previously vasa), wnt5b, cyp1a and samhd1. Expression of star, cyp11a2 and cyp1a in males was also responsive to ATR exposure. Overall, our results confirm that early development is a sensitive window for estrogenic endocrine disruption in LMB and are consistent with the hypothesis that ATR exposure induces some estrogenic responses in the developing gonad. However, ATR-specific and EE2-specific responses were also observed.

2-Ethylhexyl Diphenyl Phosphate and Its Hydroxylated Metabolites are Anti-androgenic and Cause Adverse Reproductive Outcomes in Male Japanese Medaka (Oryzias latipes).

Although high concentrations of 2-ethylhexyl diphenyl phosphate (EHDPP) have been detected in wild fish, its reproductive toxicity in fish remains unclear. In this study, we for the first time observed that EHDPP elicited androgen receptor (AR) antagonistic activity with a 50% inhibitory concentration of 37.5 μM. 2-Ethyl-5-hydroxyhexyl diphenyl phosphate was proved to be the dominant metabolite of EHDPP in Japanese medaka and elicited 3.1-fold stronger AR antagonistic activity than that of EHDPP. Medaka larvae (0-day post hatching) were exposed to EHDPP for 100 days, and intersex was observed in males from all exposure groups with significantly increased incidence (13.5-48.6%). 17β-E2 was promoted at 104 ng/L, and androgens were suppressed at 434 ng/L, which account for the intersex incidence in the high-exposure groups but do not explain the significant incidence of intersex in the 29.9 ng/L exposure group. The AR antagonistic activity of EHDPP and its metabolites must therefore play a key role in intersex incidence. EHDPP also significantly (p < 0.05) repressed reproductive behaviors of males in the 434 ng/L group and decreased fertility in high-exposure groups compared with the control. All the adverse outcomes were observed under environmentally relevant concentrations, implying that EHDPP poses an ecological risk for wild fish populations.

Chronic simultaneous exposure of common carp (Cyprinus carpio) from embryonic to juvenile stage to drospirenone and gestodene at low ng/L level caused intersex.

Synthetic progestins are emerging contaminants of the aquatic environment with endocrine disrupting potential. The main aim of the present study was to investigate the effects of the synthetic progestins gestodene, and drospirenone on sex differentiation in common carp (Cyprinus carpio) by histological analysis. To gain insights into the mechanisms behind the observations from the in vivo experiment on sex differentiation, we analyzed expression of genes involved in hypothalamus-pituitary-gonad (HPG) and hypothalamus-pituitary-thyroid (HPT) axes, histology of hepatopancreas, and in vitro bioassays. Carp were continuously exposed to concentrations of 2 ng/L of single progestins (gestodene or drospirenone) or to their mixture at concentration 2 ng/L of each. The exposure started 24 h after fertilization of eggs and concluded 160 days post-hatching. Our results showed that exposure of common carp to a binary mixture of drospirenone and gestodene caused increased incidence of intersex (32%) when compared to clean water and solvent control groups (both 3%). Intersex most probably was induced by a combination of multiple modes of action of the studied substances, namely anti-gonadotropic activity, interference with androgen receptor, and potentially also with HPT axis or estrogen receptor.

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.

Chronic nitrate exposure alters reproductive physiology in fathead minnows

Nitrate is a ubiquitous aquatic pollutant that is commonly associated with eutrophication and dead zones in estuaries around the world. At high concentrations nitrate is toxic to aquatic life but at environmental concentrations it has also been purported as an endocrine disruptor in fish. To investigate the potential for nitrate to cause endocrine disruption in fish, we conducted a lifecycle study with fathead minnows (Pimephales promelas) exposed to nitrate (0, 11.3, and 56.5 mg/L (total nitrate-nitrogen (NO3-N)) from <24 h post hatch to sexual maturity (209 days). Body mass, condition factor, gonadal somatic index (GSI), incidence of intersex, and vitellogenin induction were determined in mature male and female fish and plasma 11-keto testosterone (11-KT) was measured in males only. In nitrate-exposed males both 11-KT and vitellogenin were significantly induced when compared with controls. No significant differences occurred for body mass, condition factor, or GSI among males and intersex was not observed in any of the nitrate treatments. Nitrate-exposed females also had significant increases in vitellogenin compared to controls but no significant differences for mass, condition factor, or GSI were observed in nitrate exposed groups. Estradiol was used as a positive control for vitellogenin induction. Our findings suggest that environmentally relevant nitrate levels may disrupt steroid hormone synthesis and/or metabolism in male and female fish and may have implications for fish reproduction, watershed management, and regulation of nutrient pollution.

Endocrine-disruptor molecular responses, occurrence of intersex and gonado-histopathological changes in tilapia species from a tropical freshwater dam (Awba Dam) in Ibadan, Nigeria

In the present study, the occurrence of endocrine disruptive responses in Tilapia species from Awba Dam has been investigated, and compared to a reference site (Modete Dam). The Awba Dam is a recipient of effluents from University of Ibadan (Nigeria) and several other anthropogenic sources. A total of 132 Tilapia species (Sarotherodon malenotheron (n=57 and 32, males and females, respectively) and Tilapia guineensis (n=23 and 20, males and females, respectively)) were collected from June to September 2014. At the reference site, samples of adult male and female S. melanotheron (48 males and 47 females) and T. guineensis (84 males and 27 females) were collected. Gonads were morphologically and histologically examined and gonadosomatic index (GSI) was calculated. Hepatic mRNA transcriptions of vitellogenin (Vtg) and zona radiata protein (Zrp) genes were analyzed using validated RT-qPCR. Significant increase in Vtg and Zrp transcripts were observed in male tilapias from Awba Dam, compared to males from the reference site. In addition, male tilapias from Awba Dam produced significantly higher Vtg and Zrp mRNA, compared to females in June and July. However, at the natural peak spawning period in August and September, females produced, significantly higher Vtg and Zrp mRNA, compared to males. Fish gonads revealed varying incidence of intersex with a striking presence of two (2) pairs of testes and a pair of ovary in S. melanotheron from Awba Dam. The entire fish population examined at Awba Dam showed a high prevalence of intersex (34.8%), involving phenotypic males and females of both species. Analysis of sediment contaminant levels revealed that As, Cd, Pb, Hg and Ni (heavy metals), monobutyltin cation, 4-iso-nonyphenol and PCB congeners (138, 153 and 180) were significantly higher in Awba Dam, compared to the reference site. Principal component analysis (PCA) showed that fish variables were positively correlated with sediment contaminant burden at Awba Dam, indicating that the observed endocrine disruptive responses are associated with contaminant concentrations. Overall, the occurrence of intersex and elevated expressions of Vtg and Zrp in male fish, suggest that the measured contaminants were eliciting severe endocrine disruptive effects in Awba Dam biota, which is an important source of domestic water supply and fisheries for the University of Ibadan community.

Whole organism responses and intersex severity in rainbow darter (Etheostoma caeruleum) following exposures to municipal wastewater in the Grand River basin, ON, Canada. Part A

Municipal wastewater effluents (MWWEs) contain anthropogenic substances that can exhibit endocrine-disrupting activity. These complex mixtures have been observed to exert adverse effects on fish. Rainbow darter (Etheostoma caeruleum, RBD) is a small benthic fish that is widespread throughout the Grand River, Ontario, Canada, and has been previously shown to be adversely affected by MWWE exposure in this watershed. The objectives of this study were to quantify biological responses in this sentinel species and intersex severity in male fish, in relation to the area of urbanization. It focused on RBD populations adjacent to wastewater outfalls in the Grand River watershed. In May 2011, nine sites across the urban gradient were selected to evaluate the impact of MWWEs. Endpoints for energy storage (i.e. condition factor, k; liversomatic index, LSI) as well as reproductive endpoints (i.e. gonadosomatic index, GSI; gonad development, hormone production), and intersex were assessed in the fish. Rainbow darter showed a high incidence of intersex downstream of the wastewater outfalls, especially below the largest treatment plant outfall at Kitchener (∼85%). We applied an intersex index (score from 0 to 7) that considers the number of eggs within the testis and the stage of maturation of the egg. RBD exposed downstream of the largest wastewater outfall at Kitchener had a score of 3.81±0.37 compared to upstream to the urban areas where there were no intersex males found other than a single individual with a score 1 (average intersex score of site 0.06±0.06). In addition, several fish associated with the Kitchener outfall had macroscopic vitellogenic eggs in the testes (intersex scores 5 and 6). The sub-population of fish located at the wastewater outfall also showed a tendency towards skewed sex ratios (greater proportion of females to male fish) compared to the population at the reference sites. Male fish inhabiting the urbanized area of the Grand River also showed reduced levels of testosterone (T) and 11-ketotestosterone (11KT). Intersex males had the lower levels of 11KT relative to the upstream reference fish but could not be distinguished from normal males collected at the exposed sites. Despite the high levels of intersex at these sites, no relationships were evident among intersex severity and other measured endpoints such as GSI, LSI or in vitro steroid production. The effects observed appear to be associated with urbanization and exposure to treated MWWEs in the watershed. Although intersex incidence and severity was a very good indicator of wastewater exposure, intersex could not be directly linked to other effects in this wild population. The effects of MWWEs on transcriptional changes in adult RBD exposed to the effluents are reported in the corresponding report (Part B).

Survey of Intersex Largemouth Bass from Impoundments in Georgia USA

AbstractIntersex fish are increasingly being reported worldwide, primarily in rivers that receive treated wastewater, but few studies have investigated intersex in waters that do not receive wastewater. In a recent reconnaissance survey of intersex fish in North America, a high rate of intersex was reported for Largemouth Bass Micropterus salmoides in some southeastern U.S. rivers; however, the occurrence of intersex in impoundments has not been well described, especially on a statewide scale. Therefore, our objective for this project was to survey the occurrence of intersex Largemouth Bass in a variety of impoundment habitats across Georgia. Largemouth Bass were collected from 11 impoundments without direct municipal or agricultural wastewater inputs. Gonads from all male Largemouth Bass were evaluated for the incidence and severity of the intersex condition based on presence and arrangement of testicular oocytes. Overall 48% of male Largemouth Bass collected from impoundments were intersex, which was found in 9 of the 11 impoundments. Among impoundments, incidence of intersex ranged from 0 to 82% of the males sampled and surface area of the impoundment was a significant predictor of intersex incidence. Intersex fish were smaller than normal males, but population‐level effects of intersex and causative factors of endocrine disruption in the impoundments remain unknown. The high incidence of intersex males in small impoundments demonstrates that the condition is not confined to rivers and suggests that factors other than those previously associated with intersex (i.e., municipal wastewater) may be involved.

Assessing the Continuous Impact of Tributyltin from Antifouling Paints in a Brazilian Mangrove Area Using Intersex in Littoraria angulifera (Lamarck, 1822) as Biomarker

Intersex is a sensitive biomarker of TBT exposure and effects in littorinid gastropods and described for the mangrove periwinkle Littoraria angulifera for the first time in this study. The objective was to describe the occurrence of intersex in L. angulifera, to propose the species as a sentinel organism to assess TBT contamination, and to characterize the contamination in mangroves. The study was carried out in 2009 by sampling at 20 stations near harbors and marinas and at a reference station on the coast of Espírito Santo Estate, Brazil. At the reference station, no intersex specimens were found, while at 20 sampling stations 51% of the females exhibited different degrees of intersex development, including the occurrence of functionally sterilized females. The highest incidence of intersex and greatest intersex intensities was found in areas close to marinas and shipyards indicating that vessel-related activities are still the main source of TBT contamination. L. angulifera collected from stations in areas with well-preserved mangroves was larger than specimens collected from other areas. These differences are attributed to environmental quality and not to occurrence of intersex. The results indicate that this region is still affected by TBT contamination and that L. angulifera has the required sensitivity to be used as a bioindicator.

Region-Wide Reproductive Disruption, Masculinization and Aromatase Suppression in Marine Fish Collected from a Coastal Hypoxic Zone.

One of the most dramatic global changes due to human activities over the last half century has been the marked increase in the extent of seasonal coastal hypoxia (dissolved oxygen, DO < 2 mg O2/L) which now affects over 400 regions and covers a total area of about 250,000 sq.km. The long-term impacts on marine ecosystems of this dramatic worldwide increase in coastal hypoxia are unknown. Here we show widespread reproductive disruption in Atlantic croaker collected from hypoxic sites 120 km apart in the extensive hypoxic region in the northern Gulf of Mexico, the second largest hypoxic region in the world. Gonadal growth and gamete production were impaired in croaker from hypoxic sites compared to fish from reference normoxic sites. Male germ cells were detected in 16-24% of croaker ovaries collected in the hypoxic region, but were absent in croaker ovaries from normoxic sites. In addition, the sex ratio was skewed towards males at the hypoxic sites. The masculinization and other reproductive disruptions were associated with declines in neuroendocrine function as well as ovarian and brain expression of aromatase, the enzyme that is critical for ovarian differentiation in fish. A similar incidence of intersex and decline in ovarian aromatase expression was observed in croaker after chronic laboratory exposure to hypoxia, indicating that masculinization of ovaries is a specific response to hypoxia and is due to decreased aromatase activity. The results suggest that severe reproductive impairment can occur over large coastal regions in marine fish populations exposed to seasonal hypoxia, with potential long-term impacts on population abundance. (poster)

Intersex and reproductive impairment of wild fish exposed to multiple municipal wastewater discharges

The Grand River watershed in Ontario, Canada, receives and assimilates the outflow of 29 Municipal Wastewater Effluent (MWWE) discharges which is a mixture of domestic and industrial wastes. The purpose of this study was to investigate the cumulative impact of multiple sewage discharges on populations of wild fish. In field studies, responses of fish populations and individual fish responses in terms of growth (condition factor), reproduction (in vitro sex steroid production, gonadosomatic indices, histology [cellular development and intersex]) were assessed upstream and downstream of two municipal discharges. Fish [Greenside Darters Etheostoma blennioides and Rainbow Darters E. caeruleum] collected downstream of two municipal wastewater plants had the potential to have greater growth (longer and heavier) when compared to reference fish collections regardless of sex. Fish were not assimilating additional anthropogenic resources into energy storage (increased condition, liver somatic index). Impacts on ovarian development appeared to be minor with no differences in growth, steroid production or cellular development. Sewage exposed male fish were experiencing impairment in the capacity to produce testosterone and 11-ketotestosterone in vitro, and in cellular development (GSI, intersex). Male darters of both species collected in the upstream agricultural region demonstrated no evidence of intersex whereas our urban reference sites had incidence of intersex of up to 20%. Rates of intersex were elevated downstream of both sewage discharges studied (33% and>60%, respectively). Lower rates of intersex at the intermediate sites, and then increases downstream of second sewage discharge suggests that fish populations have to potential to recover prior to exposure to the second sewage effluent. Pre-spawning darters demonstrated dramatically higher incidence of intersex in the spring at both urban reference sites (33% and 50%, respectively), and increased more so downstream of the near-field and far-field exposure sites (60% and 100%, respectively). These findings suggest that the compounds released in STP effluents have a tendency to act on the male reproductive system. These effects may become more pronounced as projected human population growth will require the aquatic environment to assimilate an increasing amount of sewage waste.

Extinction Risk of Exploited Wild Roach (Rutilus rutilus) Populations Due to Chemical Feminization

A model that assesses risks posed by feminization to wild populations of roach was developed. A population life table matrix model that considered both sexes and a newly developed fertility kinetic function was applied to calculate the intrinsic population growth rate (lambda) of roach populations where males had been feminized. The maximum sustainable yield (MSY) was used to quantify the effect of various degrees of feminization on sustainability of exploited fisheries. Risk of extinction was calculated for wild roach populations. The results of the simulations suggested that (a) In the absence of fishing pressure lambda would only be decreased 1.5-1.7% even in the presence of a 100% incidence of intersex; (b) in the presence of selective fishing, the occurrence of intersex could significantly increase the extinction risk of local roach populations; (c) the benchmark value for the severity index of intersex and sex ratio required for a sustainable population of roach were estimated to be 1.13 and 0.57, respectively. The approach presented here provides a tool to (1) understand effects of male's feminization on population dynamics; (2) assess extinction risk of wild roach populations from feminization; (3) assist environmental managers in making policy decisions relative to fishery resource conservation.

Widespread occurrence of intersex in black basses ( Micropterus 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′-DDE, p, p′-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. Few reproductive 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.

Incidence of intersex in male clams Scrobicularia plana in the Guadiana Estuary (Portugal)

Endocrine disrupting chemicals (EDCs) known to be present in the marine environment, have great potential to influence reproductive status in aquatic organisms. A widespread incidence of intersex linked with EDCs exposure has been reported in molluscs and fish species. The species Scrobicularia plana is inherently gonochoristic; nonetheless, the feminisation of male clams was described in estuaries from the United Kingdom. Thus, this study reports the presence of oocytes in the testis of male clams of S. plana collected in the Guadiana River (Portugal). Samples were collected between June and September (2007), and gonad tissue analysed by light microscopy. Intersex with several degrees of intensity was detected during the first three months of sampling, in which the percentage of male clams affected varied between 5.9, 58.3 and 71.4%. Little is known about the causes of intersex in S. plana, being the urban, industrial and agricultural discharges possible major sources of induction. In the Guadiana River, the presence of several EDCs may be directly linked to the incidence of the intersex condition detected in Scrobicularia plana.

Amphipod intersex, metals and latitude: A perspective

Intersexuality has been widely reported in crustaceans with several mechanisms being directly or indirectly held responsible for its occurrence, amongst which pollution. No mechanistic relationship between metals and intersex has ever been established. Also the incidence of intersex in populations of the same invertebrate species in a latitudinal gradient has never been studied so far. Three populations (Iceland, Scotland and Portugal) of the amphipod Echinogammarus marinus were scrutinized. Intersex females from Iceland registered the highest fecundity loss. Only in Scottish samples females with two genital papillae and males with only one genital papillae were observed. Nevertheless, water, biota and sediment samples pointed to equivalent metal levels, and in consonance the prevalence of intersex was not significantly different between locations. An unequivocal relationship between metal presence and intersex induction cannot be presented, but our results advocate the potential role of metals as a direct cause of intersexuality in E. marinus.

Effects of the fungicide prochloraz on the sexual development of zebrafish ( Danio rerio)

Some chemicals have the potential to adversely affect sexual development through multiple endocrine actions. Prochloraz is an imidazole fungicide that displays diverse mechanisms of action, including inhibition of aromatase activity, inhibition of androgen synthesis, and antagonism of the androgen receptor. The objective of this study was to assess the effects of prochloraz on the sexual development of zebrafish ( Danio rerio) in the Fish Sexual Development Test (FSDT) proposed as an OECD test guideline for detection of endocrine disruptors. Zebrafish were exposed to prochloraz (0, 16, 64 or 202 μg/L) for 60 days from 24 h post fertilization. Fish exposed to 202 μg/L prochloraz showed an increased proportion of males. Furthermore, the incidence of intersex and the stages of the gonads were altered in the treated fish compared to the control fish. A significant vitellogenin decrease was observed in both female and male zebrafish at an exposure concentration of 202 μg/L prochloraz. However, in the male fish, significantly increased vitellogenin concentrations were observed in the groups exposed to 16 or 64 μg/L prochloraz. This study serves as a part of the validation of the FSDT and indicates that the FSDT is suitable in detecting compounds with multiple endocrine actions. This is of importance in the assessment of the potential risk of existing and new chemicals.

Male Coho Salmon (Oncorhynchus kisutch) Exposed to a Time-Course of Urban Sewage Effluent Exhibit a Sporadic Low Incidence of Sex Reversal and Intersex

Abstract Developing coho salmon alevin (Oncorhynchus kisutch) were exposed to concentrations of 1 to 10% municipal wastewater effluent (MWWE) during the labile period of sexual differentiation. MWWE was collected at regular time intervals from a municipal wastewater treatment plant during a 4-week period that coincided with the latter part of the spawning season of coho salmon. The salmon were exposed, from just prior to hatch until 28 d post hatch, to MWWE collected at 3 a.m., 8 a.m., 3 p.m. or 8 p.m. Subsequent gonadal development was assessed by histology and compared to genetic sex as determined by Y-chromosomal DNA markers. The MWWE (analyzed using GC-HRMS) had relatively high concentrations of steroidal estrogens and other endocrine active substances at some time points during the exposure period. A low incidence of intersex and sex reversal was noted in the coho salmon exposed to MWWE collected at only two time points; 3 a.m. (1% MWWE) and 8 p.m. (10% MWWE), and ≤19% of the male fish in these exposure groups were affected. The coho salmon utilized were of wild genetic backgrounds and thus the sporadic nature of the response observed may be due to a significant genetic influence on the ability to respond to exogenous hormones at this early stage of development.