The effects of pH on fluoxetine in Japanese medaka (Oryzias latipes): acute toxicity in fish larvae and bioaccumulation in juvenile fish.

Recent detection of fluoxetine in the aquatic environment and fish suggests a possibly high accumulation of fluoxetine; however, no report is available on the bioaccumulation of fluoxetine in aquatic organisms. Since bioaccumulation of fluoxetine was probably dependent on pH near the pK(a) value of 10.1, experiments were conducted approximately at pH 7, 8, and 9. Distribution coefficients between 1-octanol and water (D(ow)), and those between synthetic membrane vesicles (liposomes) and water (D(lip-wat)) were determined at pH 7, 8, and 9. The D(ow) and D(lip-wat) values increased significantly with increasing pH. Acute toxicity tests were performed using Japanese medaka (Oryzias latipes) prior to the bioaccumulation test, and 96-h LC(50) values were 5.5, 1.3, and 0.20mgl(-1) at pH 7, 8, and 9, respectively. In the bioaccumulation test, concentrations of fluoxetine and its major metabolite, norfluoxetine, in the fish body and liver were measured. The bioconcentration factors (BCF) of fluoxetine for Japanese medaka were 8.8, 3.0x10, and 2.6x10(2) in the body and 3.3x10(2), 5.8x10(2), and 3.1x10(3) in the liver at pH 7, 8, and 9, respectively. The BCF values were lower at pH 7 and higher at pH 9 mainly because of the increase in nonionized species with significantly higher hydrophobicity than the ionized species at pH values closer to pK(a). A similar trend was obtained for the concentration of norfluoxetine in the fish but the pseudo-BCF values (the ratio of the norfluoxetine concentration in the fish and the fluoxetine concentration in test water) were higher than the BCF value of fluoxetine at all pH conditions.

Initial ecological risk assessment of eight selected human pharmaceuticals in Japan.

Eight pharmaceuticals were selected on the basis of their domestic consumption in Japan, the excretion ratio of the parent compound and the frequency of detection in the aquatic environment or wastewater treatment plant effluent. Toxicity tests on these pharmaceuticals were conducted using Japanese medaka (Oryzias latipes), daphnia (Daphnia magna), and green algae (Psuedokirchneriella subcapitata). Predicted no effect concentration (PNEC) was calculated using lethal or effect concentration 50 (LC50 or EC50) values and no effect concentration (NOEC) obtained in the toxicity tests for these compounds. Predicted environmental concentration (PEC) was also calculated from annual consumption, the excretion rate of the parent compound, and removal rate in the preliminary batch activated sludge treatment performed in this study. Maximum concentrations found in the aquatic environment or sewage effluent in Japan or foreign countries were also used for another calculation of PEC. Initial risk assessment on the selected pharmaceuticals was performed using the PEC/PNEC ratio. The results of initial risk assessment on the eight selected pharmaceuticals suggest neither urgent nor severe concern for the ecological risk of these compounds, but further study needs to be conducted using chronic toxicity tests, including reproduction inhibition and endocrine disruption assessments.

Preliminary ecological risk assessment of butylparaben and benzylparaben -1. Removal efficiency in wastewater treatment, acute/chronic toxicity for aquatic organisms, and effects on medaka gene expression.

Butylparaben and benzylparaben, used as preservatives mainly in cosmetic products, have recently been found to be weakly estrogenic. Batch activated-sludge treatment and batch chlorination were carried out to roughly determine the removal efficiency of a wastewater treatment plant. Combining the removal efficiency with the estimated annual consumption and the unaltered excretion ratio, the maximum predicted environmental concentration (PEC) was estimated. Conventional acute/chronic toxicity tests were conducted using Japanese medaka (Oryzias latipes), daphnia (Daphnia magna), and green algae (Pseudokirchneriella subcapitata) for n-butylparaben, i-butylparaben, and benzylparaben. Medaka vitellogenin assays were also conducted for the three compounds and DNA microarray analysis was carried out to examine the effects of benzylparaben on gene expression. The plasma vitellogenin concentration of male medaka increased for concentrations of 200, 100, and 100 microg L(-1) n-butylparaben, i-butylparaben, and benzylparaben for 14 days, respectively, while the expression levels of genes encoding proteins such as p53, cytochrome P450 3A40, and choriogenin-L increased for concentrations higher than 4 microg L(-1) of benzylparaben. Furthermore, the predicted no-effect concentration (PNEC) was calculated using the lethal or effect concentration 50 (LC50 or EC50) values and no-effect concentrations (NOECs) obtained in the toxicity tests for these compounds. The maximum concentrations found in the aquatic environment or sewage effluent (MEC eff) were used to carry out preliminary environmental risk assessment. The calculated MEC/PNEC ratio suggests the necessity of further study such as a more detailed large-scale monitoring and chronic toxicity tests including reproduction inhibition and endocrine disruption.

Feminization of Japanese medaka (Oryzias latipes) exposed to 17beta-estradiol: effect of exposure period on spawning performance in sex-transformed females.

Two groups of Japanese medaka (Oryzias latipes) were exposed to 17beta-estradiol (E2: 150ng/L, nominal concentration) for either a short-term exposure (STE: 0-31 days after fertilization (daf); egg-larval period) or a long-term exposure period (LTE: 0-81 daf; egg-adult period) and their subsequent spawning performance was compared in terms of fecundity, spawning time, and fertility. Most genetic males were transformed to phenotypic females by E2 following both short-term and long-term exposure, but spawning performance and gonad somatic index (GSI) of sex-transformed females (XY females) following long-term exposure were lower than those of sex-transformed females following short-term exposure and those of normal females (XX) in the control group. Sex-transformed females in the STE group and normal females possessed mature ovary, whereas most of the sex-transformed females in the LTE group possessed immature ovary, with most oocytes being in the pre-vitellogenic phase. Moreover, the chromosome types of first filial generation delivered from sex-transformed female in STE group composed with 51.9% as XY, 18.5% as YY, and 29.6% as XX. From these results, it seems that exposure to E2 until the end of the larval period produces sex-transformed medaka with high reproductive ability, similar to normal females, but longer exposure to E2 may inhibit sexual maturation in the sex-transformed female.

Feminization of Japanese medaka (Oryzias latipes) exposed to 17beta-estradiol: formation of testis-ova and sex-transformation during early-ontogeny.

Gonad histological changes were examined in Japanese medaka exposed to 17beta-estradiol (E2) during early-life stages. Two experiments were conducted at different concentrations of E2 (33.5 and 140.6 ng/L, mean value of measurement) and larvae and juveniles were observed for histological changes in the gonad. Differentiation of ovary and testis in control fish was apparent 12 days post-hatch (dph). At 12 dph, normal testes were observed in male fish that had been exposed to 33.5 ng/L E2, but at 14 and 20 dph, testis-ova was recognized in male fish. Male fish exposed to 140.6 ng/L E2 had testis-ova at 12 dph and gradual transformation to ovary was observed in male fish until 20 dph. In both experiments, the ovarian tissue in testis of male fish exposed to E2 was frequently distributed along the central transverse axis of the gonad, expanding into the transverse axis. The results indicated that 17beta-estradiol can induce testis-ova in male medaka during the larval period and sex-transformation is more frequent at higher (140.6 ng/L) than lower concentrations (33.5 ng/L) of estradiol. The results also demonstrated that testis-ova first appear in the central area of the transverse axis of testis.

Bioconcentration factor of relatively low concentrations of chlorophenols in Japanese medaka.

Bioconcentration factors (BCF) for pentachlorophenol (PCP) and 2,4-dichlorophenol (2,4-DCP) in Japanese medaka (Oryzias latipes) were determined at five different concentrations of the chemicals, between 0.1 and 10 microg/l (PCP), 0.3 and 30 microg/l (2,4-DCP), in the ambient water. Medaka were exposed to each chemicals in a continuous-flow system during the embryonic development period and 60 days after hatching from eggs collected in the laboratory. Both the exposure time and the aqueous concentrations are much more realistic and closer to natural aquatic environments than those used in conventional BCF studies. The BCF values of PCP were from (4.9+/-2.8)x10(3) at the aqueous concentration of 0.074+/-0.028 microg/l to (2.1+/-1.4)x10(3) at 9.70+/-0.56 microg/l. The BCF value of 2,4-DCP were from (3.4+/-3.0)x10(2) at 0.235+/-0.060 microg/l to 92+/-27 at 27.3+/-1.6 microg/l. Generally, BCF values increased as the aqueous concentrations of PCP or 2,4-DCP decreased. This finding suggests that a relatively low and realistic aqueous concentration of these compounds is necessary to more accurately determine their BCF values in natural aquatic environments. Conventional BCF experiments at higher aqueous concentrations may underestimate the BCF values.

Seasonal changes in sex ratio, maturation, and size composition of fresh water snail, Sinotaia quadrata histrica, in Lake Kasumigaura.

Sinotaia quadrata histrica is a fresh water viviparous snail distributing from the Kanto region to Kyushu Island, Japan. About 7000 snails were collected in Lake Kasumigaura (L. Nishiura and L. Kitaura) in 2001 and 2002, and the sex ratio, maturity in terms of the gonad-somatic index (GSI) and operculum diameter were determined. The total female proportion was 55.2% in 2001, 53.0% in 2002 in L. Nishiura, and that of L. Kitaura was 60.4% in 2002. Comparing the season, the female proportion was the highest during early summer in both 2001 (59.6%, July in L. Nishiura) and 2002 (61.6%, June in L. Nishiura, 65.8%, July in L. Kitaura). The GSI of females in L. Nishiura significantly increased from April to May and significantly decreased from June to August. The GSI of males was higher in spring, but significantly lower from June to August. The mean female operculum diameter was consistently larger than that of males for each month and year, and a particularly significant difference was found between females and males from April to August 2001, and from April to September 2002. The number of resting zones on the operculum correlates with the operculum diameter and the female proportion was larger in the snails, which have a high number of resting zones, suggesting a sex-dependent difference in age composition. This study estimated that the sex ratio and seasonal maturation of S. quadrata histrica and the sex-dependent difference in age composition might contribute to the population structure in L. Kasumigaura.