Male Japanese Medaka Research Articles

The effect of particle size on the bioavailability of estrogenic chemicals from sediments

The effects of particle size on the bioavailability of estrogenic chemicals in the sediments from the Yeongsan River and its tributaries in South Korea were evaluated for 2006 and 2007. Samples for chemical analysis and bioassays were collected from 6 sampling sites during both dry and rainy seasons. The pore water of the sediment samples was extracted, and estrogenic chemicals were eluted using a liquid–liquid extraction (LLE) method. Concentrations of 4-nonylphenol (NP), 4- tert-octylphenol (OP), bisphenol A (BPA), estrone (E1), 17β-estradiol (E2), 17α-ethynylestradiol (EE2) and genistein (Gen) were analyzed by gas chromatography–mass spectrometry (GC–MS). To evaluate bioavailability, hepatic vitellogenin (Vtg) concentrations of male Japanese medaka were measured after exposure to the sediment or its fractions for 7 d. NP, BPA and E2 were detected in all the sediment sample extracts from the Yeongsan River and its tributaries. The concentrations of NP in the sedimentary samples ranged from 60 to 400 ng g −1 on a dry weight basis. Similarly, OP and E2 were detected in nearly all the sediment extracts, with concentrations of 13 and 26 ng g −1, respectively. According to the bioassay test results, all the sediment samples significantly induced Vtg in male fish after 7 d of exposure. Fractionation of sediments into different size-classes (i.e., particle size >1 μm, particle size <1 μm) eliminated bioavailable estrogenic activity, but fine particles of less than 1 μm in size increased the absorption of E2 from E2-amended sediment particle fractions. Consequently, the study suggested that the presence of particles and its interaction in the water environment might change the bioavailability of estrogenic chemicals.

The Effect of Estrogens, River Water, and Treated Wastewater on Vitellogenin Induction of Japanese Medaka1

Abstract: The objective of this study is to evaluate the effect of estrogens on fish endocrine disruption in river water and treated wastewater. Endogenous estrogen estrone (E1), 17β‐estradiol (E2), river water, and treated wastewater were used for exposure tests on male Japanese medaka. Vitellogenin induction in male medaka was regarded as the endpoint of endocrine disruption. The effective concentrations of E1 and E2 on vitellogenin induction in medaka were evaluated by breeding medaka for 14 days in tanks with various concentrations of E1 or E2. Vitellogenin induction increased with elapse of time during exposure, with higher estrogen concentrations causing greater vitellogenin induction. According to the test results, the lowest observed effect concentrations (LOECs) of E1 and E2 were estimated to be 31.6 and 5.0 ng/l, respectively; and the E2 equivalent (EEQ) LOEC value of E1 was 5.2 ng‐E2/l, derived by multiplying the relative potency of E1 by that of E2 as estimated in this study. In continuous exposure tests using river water or treated wastewater, the estrogenic activity, an index of total estrogenic potential measured by yeast screen assay and expressed as EEQ, varied widely during the exposure tests, and significant vitellogenin induction was observed after several days of high levels of estrogenic activity in water. Vitellogenin concentration tended to significantly increase if EEQ exceeded the level of 5 ng‐E2/l. The threshold value was substantially the same as the results for E1 and E2 exposure tests of medaka. Consequently, EEQ over 5 ng‐E2/l was revealed to have the potential to cause endocrine disruption of male medaka. As estrogenic activity exceeding 5 ng‐E2/l was observed in some rivers in Japan, total estrogenic potential needs to be further decreased at wastewater treatment plants to prevent fish endocrine disruption.

Interactions between estrogenic chemicals in binary mixtures investigated using vitellogenin induction and factorial analysis

Considerable progress has been made in assessing the combined effects of chemicals, but the effect of mixtures remains one of the most daunting challenges in environmental toxicology. In this study, the effects of binary mixtures of estrogenic chemicals were investigated using plasma vitellogenin (VTG) induction in male adult Japanese medaka ( Oryzias latipes) as the endpoint. We focused on whether or not the factorial design using rigorous statistical methods was appropriate for the identification of possible interactions. The estrogenic activities of 17β-estradiol (E2), 4- tert-nonylphenol (NP) and bisphenol A (BPA) were investigated in medaka following 14 d of exposure. Although all chemicals induced concentration-dependent increases in plasma VTG, a complete concentration–response curve was obtained only for E2, while partial curves were obtained for NP and BPA. Therefore, a 3×3 factorial design was employed to identify both the individual and interactive effects. A significant difference in mean plasma VTG induction was found when any chemical was considered alone. However, no significant interactions between E2 and NP, E2 and BPA, and NP and BPA were demonstrated. These results suggest that these three model estrogenic chemicals share a common mechanism for inducing VTG synthesis, and that no interactions occur when they act in combination. We also demonstrated the ability of this experimental design to detect interactions between binary mixtures, which will allow the assessment of biological effects of more complex mixtures in future research.

Expression Levels of Hormone Receptors and Bone Morphogenic Protein in Fins of Medaka

In the genus Oryzias, the morphologies of the dorsal and anal fins are typical secondary sex characters. In the Japanese medaka (Oryzias latipes) and Thai medaka (Oryzias minutillus), androgen receptor (AR) expression levels in the dorsal, anal, and pectoral fins were higher in males than in females. Conversely, in both species estrogen receptor (ER) beta expression levels in the dorsal and anal fins were higher in females than in males. AR and ERbeta expression levels in the dorsal and anal fins of sex-undeterminable individuals of Thai medaka were intermediate between those in normal male and female Thai medaka. There was no difference in the bone morphogenic protein (Bmp) 2b expression level between male and female Japanese medaka. In contrast, the Bmp2b expression level in the dorsal fin of sex-undeterminable individuals was lower than in normal male and female Thai medaka. It is thus clear that androgen and estrogen regulate the sex-dependent characters of fin morphology in both Oryzias species. In sex-undeterminable individuals of Thai medaka, the low levels of Bmp2b expression in the dorsal fin are evidence that androgen and estrogen are necessary for adequate expression of Bmp2b in the normal development of at least the dorsal fin.

Levels of Vitellogenin in Male Japanese Medaka (Oryzias latipes) Exposed to the Chemotherapeutics: Oxytetracycline, Romet-30®, and Copper Sulfate

ABSTRACT Japanese medaka (Oryzias latipes) were exposed to oxytetracycline and Romet-30® at daily doses via feed of 0.06 mg, 0.11 mg, and 0.28 mg per gram body weight for 11 days, and at daily doses of 0.05 mg, 0.10 mg, and 0.15 mg per gram body weight for 5 days, respectively. In addition, fish were exposed to copper sulfate in their aquaria water at 1.85 mg/L for 1 h each day (over a span of 1 day, 7 consecutive days, or 1 day in every 3 to 5 days for a total of seven doses). Vitellogenin levels were measured in fish using a sandwich enzyme-linked immunosorbent assay (ELISA). Vitellogenin levels were not induced from exposure to either oxytetracycline or Romet-30® (P > 0.05); however, copper sulfate for 1 h on 1 day showed a decrease in vitellogenin from the control fish and other treatment groups (P = 0.007). Under these test conditions, oxytetracycline, Romet-30, and copper sulfate were not found to act in medaka as estrogenic compounds by inducing vitellogenin.

Estrogenic activity and reproductive effects of the UV-filter oxybenzone (2-hydroxy-4-methoxyphenyl-methanone) in fish

Previous studies in extracts of sediments surrounding municipal outfalls off the coast of California, USA and effluents of New York City, NY, USA indicated the UV-filtering agent, oxybenzone (CAS# 131-57-7; benzophenone-3) as a potential estrogen. The effects of oxybenzone on estrogenic activity and reproduction were evaluated using a 14-day juvenile rainbow trout assay for plasma vitellogenin and a subsequent 21-day Japanese medaka reproduction assay. Significant induction of vitellogenin was observed in the rainbow trout at the 1000 μg/L nominal concentration (749 μg/L median measured value) of oxybenzone which was approximately 75 times greater than the concentrations observed in previous wastewater effluent. Vitellogenin induction was also observed in the 1000 μg/L nominal concentration (620 μg/L median measured) of oxybenzone in male Japanese medaka ( Oryzias latipes) after 21 days of exposure. The number of eggs produced per female per day exposed to the same concentration (620 μg/L) were significantly lower after 7 days, but returned to control values after 21 days. Fertilized eggs were then monitored for 20 days to assess hatching success. The overall percentage of fertilized eggs collected during the 21-day exposure that hatched was significantly lower in the 620 μg/L oxybenzone concentration. There was also a temporal effect at this concentration as egg viability (percentage of fertilized eggs that hatched) was diminished 13–15 days after eggs were collected. All three oxybenzone concentrations (16, 132, and 620 μg/L) and the 50 ng/L estradiol positive control showed reduced hatching of eggs at day 15, and the 132 and 620 μg/L oxybenzone concentrations diminished the percentage of eggs that hatched on days 13–15. These data indicate that the UV-filter oxybenzone alters endocrine or reproduction endpoints in two fish species, but at concentrations significantly higher than those measured in the environment.

Chlortetracycline Caused Vitellogenin Induction at Male Japanese Medaka (Oryzias latipes)

This investigation was intended to find out the estrogenic effect of chlortetracycline (CTC) on vitellogenin induction in adult male Japanese medaka (Oryzias latipes). Vitellogenin (Vtg) produced in male fish has been used to as one of a biomarker of endocrine disrupters. The positive control was <TEX>$17{\beta}-estradiol$</TEX> (E2) that induced Vtg in male fish. As a result, male and female fish were exposed to 0.1, 1, 10 and 100 ppm of CTC. Western blot results showed approximately 205 kDa, that is similar to myosin at high molecular weight range Sigma maker. Vtg band was showed fainted to 10 and 100ppm for chlortetracycline. Vtg concentration of CTC was qunatified by total protein quantification and ELISA. Exposure of the male fish to CTC of 0.1, 1, 10 and 100 ppm produced Vtg concentrations of 0.24, 0.12, 7.61 and 40.02%o, respectively, that value was elevated than control male fish (0.14%o). CTC exerted as a Vtg inducer in male fish from 10 ppm, but it was a reducer in female fish from 0.1 ppm level. The results say that vitellogenin induction patterns alter in male medaka treated with CTC, and that CTC may caused endocrine disruption in fish.

Comparative sensitivity in Chinese rare minnow (Gobiocypris rarus) and Japanese Medaka (Oryzias latipes) exposed to ethinylestradiol

Morphological and biochemical indices, including hepatosomatic index, gonadosomatic index, and vitellogenin (VTG) induction, were compared between two teleostean species to determine their relative sensitivity of exposure to 17-α -ethinylestradiol (EE2). Chinese rare minnow (Gobiocypris rarus) and Japanese medaka (Oryzias latipes) were exposed to aqueous concentrations of EE2 ranging from 0.2 to 100 ng/L for 21 days. A stronger liver response to EE2 in Chinese rare minnow was observed than that in Japanese medaka. In Japanese medaka, significantly increased male GSI could be observed at 50 and 100 ngEE2/L. The semi-quantitative estimation by integrated optic density following electrophoresis showed that exposure of male Chinese rare minnow and Japanese medaka to EE2 resulted in significant (P < 0.05) induction of VTG at 0.2 and 2 ngEE2/l, respectively. It indicated that the sensitivity of Chinese rare minnow to the stress of EE2 is about 10-fold greater than that of Japanese medaka.

2,3,7,8-Tetrachlorodibenzo-p-Dioxin (TCDD) Induces Organ- Specific Differential Gene Expression in Male Japanese Medaka (Oryzias latipes)

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a ubiquitous environmental contaminant with well-known adverse effects in fish. In this study, we initially exploited suppression subtractive hybridization (SSH) as a screening tool to assess qualitative gene expression changes in whole brain, liver, and testis of adult male Japanese medaka (Oryzias latipes) exposed for 48 h to a single intraperitoneal-injected dose of TCDD (10 microg TCDD/kg body weight). Across these three organs, SSH identified a total of 335 unique genes. Each set of forward- and reverse-subtracted organ cDNA libraries consisted of a distinct gene list and corresponding distribution of biological processes, suggesting that transcript profiles of these libraries were highly organ-specific. Based on sequence match significance and frequencies within each set of organ libraries, genes hypothesized to be strongly responsive (42 total) within male medaka brain, liver, or testis were semi-quantitatively screened with replicate cDNA nylon membrane arrays. In addition, TCDD-treated male medaka were surveyed for gross histological analysis of brain, liver, and testis. In general, adverse histopathological changes were not observed in the brain, and glycogen depletion was observed only in the liver. However, significant histological changes occurred in the testis, and included disorganization of spermatogenesis at the testis periphery, disruption of the interstitium, Leydig cell swelling, and Sertoli cell vacuolation. Of the 42 genes screened by cDNA array analysis, cytochrome P450 1A (CYP1A) mRNA was the only transcript significantly higher in TCDD-exposed brain, whereas 12 transcripts (including CYP1A) were significantly higher in TCDD-exposed liver, and 34 transcripts were significantly lower in TCDD-exposed testis. Therefore, the degree of TCDD-induced alterations observed in each organ at a gross histological level corresponded well with the number and ontology of gene transcripts affected on the array. Based on real-time reverse transcription polymerase chain reaction (RT-PCR), relative CYP1A (but not AHR1) transcript levels were confirmed to be significantly higher in TCDD-treated brain and liver. However, CYP1A was not significantly induced in TCDD-exposed testis, suggesting that gene expression and histopathological responses observed in the testis at 48 h may be CYP1A-independent. Based on these data, unique liver-specific and testis-specific mRNA-level targets in male medaka were identified as promising biomarkers of acute TCDD-induced toxicity.

The effect of bisphenol A and chlorinated derivatives of bisphenol A on the level of serum vitellogenin in Japanese medaka (Oryzias latipes)

2,2-bis (4-hydroxyphenyl) propane or Bisphenol A (BPA), has been reported to behave as an endocrine disrupter below acute toxic levels, and is widely present in the water environment. Although BPA is easily chlorinated, very little is reported on the effect of chlorinated BPA to the aquatic organisms. In this study, the estrogenic activities of BPA and its chlorinated derivatives were evaluated by the induction of vitellogenin (VTG) in the serum of mature male Japanese medaka. In addition, the effect of sodium hypochlorite on the decomposition of BPA was tested. The relative potencies of estrogenic activities of chlorinated BPA descended in the order 3,3'-diCIBPA>BPA> or =3-CIBPA>3,3',5-triCIBPA, and no estrogenic activity was observed in 3,3',5,5'-tetraCIBPA. Lowest Observed Effect Concentration (LOEC) and No Observed Effect Concentration (NOEC) for both 3-CIBPA and 3,3'-diCIBPA were 500 microg/L and 200 microg/L, respectively. LOEC for 3,3',5-triCIBPA was >500 microg/L. When BPA was reacted with sodium hypochlorite (24 hours; residual chlorine at 1 ppm), however, complete decomposition of BPA and its chlorinated derivatives was observed. The decrease in BPA and its chlorinated derivatives paralleled the decrease in estrogenic potency evaluated by the induction of vitellogenin (VTG) in the serum of mature male Japanese medaka.

Decreased apoptosis in the forebrain of adult male medaka ( Oryzias latipes) after aqueous exposure to ethinylestradiol

Endocrine disrupting compounds (EDCs), especially those that are estrogenic, are an issue of growing concern because they may ultimately adversely affect wildlife survival. 17-β-Estradiol and its synthetic counterpart, 17-α-ethinylestradiol, two common EDCs, are associated with intersex conditions and impaired male reproductive behavior in fish. Male and female Japanese medaka ( Oryzias latipes) were exposed to 10 ng/l ethinylestradiol for 6 months. Using terminal dideoxynucleotidyl-mediated dUTP nick end-labeling (TUNEL) to quantitate cell death, we found that ethinylestradiol-exposed males had significantly fewer apoptotic cells in the forebrain compared to untreated males and exposed females. Our results show that the effects of ethinylestradiol exposure are highly variable among individuals of the same species and even within tissues of the same individual. Thus, when examining the effects of EDCs on natural populations, data from a variety of tissues should be examined and the interpretation of any effects should include consideration of tissue-specific processes.

Effects of tamoxifen, 17α-ethynylestradiol, flutamide, and methyltestosterone on plasma vitellogenin levels of male and female Japanese medaka ( Oryzias latipes)

The effects of oral administration of tamoxifen, 17α-ethynylestradiol (EE2), flutamide, and methyltestosterone (MT), on plasma vitellogenin levels of male and female medaka were investigated. Medaka were fed diets containing different concentrations of these chemicals for 7 days, and these plasma vitellogenin levels were measured. Tamoxifen increased significantly the vitellogenin levels in male, but inhibited the normal vitellogenin induction in female in the high concentration groups. EE2 increased significantly vitellogenin levels in both sexes. Flutamide increased significantly the vitellogenin levels in female, but gave no effects on male. MT inhibited the normal vitellogenin induction in female, but increased slightly vitellogenin levels in male without a clear tendency. Administration of tamoxifen, EE2, flutamide, and MT showed the different pattern in vitellogenin levels in both sexes.

Alteration of General Behavior of Male Medaka,oryzias latipes,Exposed to Tributyltin and /or Polychlorinated Biphenyls

We examined the general behavior of male Japanese medaka, Oryzias latipes, after exposed to tributyltin (TBT), polychlorinated biphenyls (PCBs), or a mixture of these chemicals at a concentration of I ~tg/g body weight daily for 3 weeks. We analyzed swimming velocity as an indicator of acute toxicity and counted the frequencies of straight swlmnting and swlmming in circles. We also calculated the entropy of the positions of the fish within the experimental chamber. Neither TBT nor PCBS nor their mixture affected the swimming velocity, suggesting that there were no acute toxic effects. However, PCBS exposure increased the frequencies of both straight and circular swimming, suggesting hyperactivity. TBT increased the positional entropy value. Both TBT and PCBS affect the general behavior of mature male medaka, although exposure to the 2 chemicals together has no significant additive behavioral effect.

Effects of cadmium on the reproductive axis of Japanese medaka ( Oryzias latipes)

Cadmium (Cd) is a ubquitous element and a significant inorganic pollutant that has previously been found to bioaccumulate in reproductive organs of fish and disrupt important endocrine processes, especially those involved in synthesis, release and metabolism of hormones. Clearly, there is potential for reproductive effects in fish populations exposed to Cd, however, few studies have investigated the non-lethal consequences of Cd in fish. To this extent, adult male and female Japanese medaka were exposed to 0–10 ppb Cd for 7 weeks. Reproductive endpoints were monitored during weeks 6 and 7 of exposure and compared to physiological responses along the hypothalamus-pituitary-gonadal (HPG) axis, including plasma vitellogenin (VTG), hepatic estrogen receptor (ER), plasma steroids, gonadal-somatic indices (GSI), and gonadal steroid release. There were no observed effects on VTG and ER by long-term Cd exposure. However, gonadal steroid release was significantly decreased in males and females at all exposure concentrations and female plasma estradiol levels were significantly altered at concentrations higher than 5 ppb Cd. Overall, responses along the HPG axis were more sensitive to Cd exposure than the reproductive and developmental endpoints, which were not affected in this study, indicating that higher level impairment in fish might be relatively protected.

Oxidative degradation of alkylphenols by horseradish peroxidase

Alkylphenols such as bisphenol A (2,2-bis(4-hydroxyphenyl)propane; BPA), p-nonylphenol ( p-NP), and p-octylphenol ( p-OP) that are known as endocrine disrupters were oxidized by horseradish ( Armoracia rusticana) peroxidase (HRP) with H 2O 2. The optimal pHs for BPA, p-NP, and p-OP were 8.0, 7.0, and 5.0, respectively. The optimal temperature for BPA was 20°C. Although BPA was rapidly degraded by HRP, its degradation depended on the concentration of HRP. Most of the oxidation products of BPA were polymers, although some 4-isopropenylphenol was produced. When male Japanese medaka ( Oryzias latipes) were exposed to BPA, vitellogenin in the blood increased. However, no increased vitellogenin was observed in medaka exposed to HRP-oxidized BPA. The enzymatic oxidation of BPA using HRP was able to eliminate its estrogen-like activity.

Suppression of sexual behavior in male Japanese medaka (Oryzias latipes) exposed to 17beta-estradiol.

Mating pairs of Japanese medaka (Oryzias latipes) received 17beta-estradiol (E2; dose, 3 and 30 microg/g body weight daily) in their diet and E2-free diet (control) for two weeks. The effect on reproductive success was assessed in the mating pairs, and sexual and nonsexual behaviors in the male were quantified. The fecundities of the E2-treated groups were reduced significantly (p < 0.05) compared with those of control, and the sexual behaviors (following, dancing, floating, and crossing) of male fish in response to prostaglandin F2alpha (PG)-injected E2-unexposed females were suppressed dramatically. While the fertility of spawned eggs and the frequencies of the nonsexual behaviors of resting and swimming in a straight line performed by male were unchanged in E2-exposed groups. Therefore, sexual behavior in male may be linked with reproductive success in mating pair of madaka exposed to E2. This assay for sexual behavior using PG may provide a useful tool for assessing the effect of endocrine-disrupting chemicals.

Increased cellular apoptosis after chronic aqueous exposure to nonylphenol and quercetin in adult medaka ( Oryzias latipes)

Increasing evidence suggests that sublethal effects of natural or xenobiotic chemicals in the environment may be mediated via the stimulation of apoptosis. To investigate whether apoptosis can be induced in fish by weakly estrogenic and androgenic chemicals, adult male Japanese medaka ( Oryzias latipes) were exposed to 100 ppb of the estrogenic alkylphenol, 4-nonylphenol, and adult female medaka were exposed to 100 ppb of the aromatase-inhibiting bioflavonoid, quercetin, for 6 weeks. Exposure to nonylphenol and quercetin had no significant effect on the length, weight or condition factors compared to solvent (acetone) controls in male or female medaka. Apoptosis was evaluated in blinded histological sections of whole medaka using terminal dideoxynucleotidyl transferase dUTP nick end labeling (TUNEL) that labels nuclei of cells containing apoptotic (fragmented) DNA. There was a six-fold greater extent of apoptosis in spermatocytes, Sertoli cells and Leydig-homologue cells, but not in spermatids of testes from nonylphenol-exposed male medaka compared to testes of solvent controls. No significant differences in the extent of apoptosis were detected in intestine, liver or kidney from the same male fish. Quercetin-treated female medaka had a significantly increased number of atretic ovarian follicles, but no significant differences in the extent of apoptosis in intestine, liver or kidney. These results suggest that nonylphenol caused testicular degeneration via increased testicular cell apoptosis, while quercetin may be ovotoxic via increased follicular atresia.

Competitor-to-resource ratio, a general formulation of operational sex ratio, as a predictor of competitive aggression in Japanese medaka (Pisces: Oryziidae)

Operational sex ratio (OSR), the number of potentially mating males divided by the number of fertilizable females, plays a central role in the theory of mating systems by predicting the intensity of intra-sexual competition and sexual selection. We introduce a general version of OSR, competitor-to-resource ratio (CRR, the number of potential competitors divided by the number of resource units), as a potential way of predicting the intensity of competition for any resource. We manipulated CRR over a broad range (0.5-8) by varying both the number of competing male Japanese medaka fish (Oryzias latipes) and the number of resources, either females or food items. We tested whether the rate of male—male aggression differed depending on resource type and whether it increased monotonically or followed a dome-shaped relationship with increasing CRR. The patterns of competitive aggression in relation to CRR did not differ significantly between resource types. In addition, the per capita rate of aggression followed a dome-shaped curve; it was low when CRR was less than one, initially increased as CRR increased, was highest at a CRR of about two, and then decreased when CRR was greater than two. However, competitor number, independent of CRR, had a significant and negative effect on rate of aggression. We suggest that CRR is a valuable predictor of the rate of competitive aggression and may be a useful concept for synthesizing ideas about resource competition and monopolization that are currently dispersed in the separate bodies of literature on mating systems, social foraging and territoriality.

Serum vitellogenin levels and reproductive impairment of male Japanese Medaka (Oryzias latipes) exposed to 4-tert-octylphenol.

The induction of synthesis of the "female" yolk precursor protein vitellogenin (VTG) in male fish by estrogenic chemicals in the environment has been demonstrated in many recent reports. However, little is known about the organismal and biological significance of this phenomenon. To examine the relationship between VTG production in male fish and reproductive impairment, adult male medaka were exposed to 4-tert-octylphenol (OP), a known environmental estrogen, in concentrations ranging from 20 to 230 ppb for 21 days, under flow-through conditions. Following exposure, male fish were mated, in the absence of OP, with unexposed females. Breeding groups composed of exposed males and control females produced about 50% fewer eggs than control groups. VTG levels in serum of male fish increased with increasing OP exposure concentration and decreased after OP exposure was discontinued. Nevertheless, significant correlations (p<0.01) were observed between VTG levels in exposed male fish and 1) OP exposure concentrations, 2) percent of fertilized eggs, and 3) survival of embryos. OP-induced VTG synthesis and reproductive impairment appear to be closely linked phenomena. Histological examination indicated spermatogenesis in OP-exposed fish was inhibited, and some exposed fish had oocytes in their testes. Finally, OP caused a significant increase in the number of abnormally developing embryos, suggesting that OP may be genotoxic as well as estrogenic.

Serum Vitellogenin Levels and Reproductive Impairment of Male Japanese Medaka (Oryzias latipes) Exposed to 4-tert-Octylphenol

The induction of synthesis of the "female" yolk precursor protein vitellogenin (VTG) in male fish by estrogenic chemicals in the environment has been demonstrated in many recent reports. However, little is known about the organismal and biological significance of this phenomenon. To examine the relationship between VTG production in male fish and reproductive impairment, adult male medaka were exposed to 4-tert-octylphenol (OP), a known environmental estrogen, in concentrations ranging from 20 to 230 ppb for 21 days, under flow-through conditions. Following exposure, male fish were mated, in the absence of OP, with unexposed females. Breeding groups composed of exposed males and control females produced about 50% fewer eggs than control groups. VTG levels in serum of male fish increased with increasing OP exposure concentration and decreased after OP exposure was discontinued. Nevertheless, significant correlations (p&lt;0.01) were observed between VTG levels in exposed male fish and 1) OP exposure concentrations, 2) percent of fertilized eggs, and 3) survival of embryos. OP-induced VTG synthesis and reproductive impairment appear to be closely linked phenomena. Histological examination indicated spermatogenesis in OP-exposed fish was inhibited, and some exposed fish had oocytes in their testes. Finally, OP caused a significant increase in the number of abnormally developing embryos, suggesting that OP may be genotoxic as well as estrogenic.