Lipophilic antioxidants and lipid peroxidation in yellow perch subjected to various anthropogenic influences along the St. Lawrence River (QC, Canada).

In Lake Saint-Pierre (LSP), the last great widening of the St. Lawrence River (province of Québec, Canada), the yellow perch has been experiencing a significant decline since the mid-1990s. The combined effect of several stressors (deterioration of habitats appropriate for reproduction and growth, invasive species and poor water quality) seems to exert considerable influence on the yellow perch population in LSP, characterized by low recruitment. To better understand possible stressor effects at the biochemical level, LSP yellow perch were compared with other sites along a gradient of increasing human influences from upstream to downstream along the St. Lawrence River. Morphometry (size, weight, circumference and Fulton's condition factor) and biomarkers associated to the peroxidation of lipids, lipophilic antioxidants (α-tocopherol and carotenoids), along with retinoids (vitamins A1and A2) and proteins were compared between sites at the larval, juvenile and adult stages. Fulton's condition factor was similar between sites for juveniles but was significantly lower in LSP adults, suggesting a weakened physiological condition. In most contaminated sites as LSP, lipid peroxidation tended to be higher in juveniles and adults whereas the lipophilic antioxidant lycopene and proteins content were lower. Retinyl esters were significantly lower for LSP fish compared to other sites, not only in larvae but also in the livers of juveniles and adults. These results are consistent with possible altered metabolism in the retinoid system of LSP yellow perch. The overall results reflect the "pressure" gradient tested, where the yellow perch from the most affected sites located downstream had impaired physiological and biochemical conditions compared to the upstream sectors.

Relationships between polybrominated diphenyl ethers and transcription and activity of type 1 deiodinase in a gull highly exposed to flame retardants.

Deca-brominated diphenyl ether (deca-BDE), composed mainly of BDE-209, is subject to usage restrictions in North America and Europe, although global action on its continued use has yet to be undertaken. Relatively large concentrations of polybrominated diphenyl ethers (PBDEs), especially BDE-209 and its higher brominated degradation products, have been reported in tissues of ring-billed gulls (Larus delawarensis) breeding near the densely populated city of Montreal (QC, Canada). There is limited knowledge of BDE-209 biotransformation and toxicokinetics in birds. Deiodinases, a class of enzymes catalyzing thyroid hormone conversion, have been suggested to be involved in BDE-209 debromination in birds. The objective of the present study was to investigate the relationships between PBDE concentrations and type 1 deiodinase (D1) transcription and in vitro activity (microsomes) in livers of Montreal-breeding ring-billed gulls. The ring-billed gulls exhibiting the highest D1 activity in liver microsomes accumulated the greatest liver concentrations of hepta-BDEs and octa-BDEs. Activity of D1 was inversely related to concentration ratios of BDE-209 to octa-BDEs and ∑hepta-BDE. An even stronger inverse relation was found between D1 activity and BDE-209 to ∑nona + octa + hepta-BDE concentration ratios. The messenger ribonucleic acid (mRNA) levels of D1 in gull livers were inversely associated with liver concentrations of ∑octa-BDE. The present study's findings suggest that D1 is potentially involved in BDE-209 biotransformation and accumulation of higher brominated PBDEs in livers of ring-billed gulls. Environ Toxicol Chem 2016;35:2215-2222. © 2016 SETAC.

Ccdc11 is a novel centriolar satellite protein essential for ciliogenesis and establishment of left-right asymmetry.

The establishment of left-right (L-R) asymmetry in vertebrates is dependent on the sensory and motile functions of cilia during embryogenesis. Mutations in CCDC11 disrupt L-R asymmetry and cause congenital heart disease in humans, yet the molecular and cellular functions of the protein remain unknown. Here we demonstrate that Ccdc11 is a novel component of centriolar satellites-cytoplasmic granules that serve as recruitment sites for proteins destined for the centrosome and cilium. Ccdc11 interacts with core components of satellites, and its loss disrupts the subcellular organization of satellite proteins and perturbs primary cilium assembly. Ccdc11 colocalizes with satellite proteins in human multiciliated tracheal epithelia, and its loss inhibits motile ciliogenesis. Similarly, depletion of CCDC11 in Xenopus embryos causes defective assembly and motility of cilia in multiciliated epidermal cells. To determine the role of CCDC11 during vertebrate development, we generated mutant alleles in zebrafish. Loss of CCDC11 leads to defective ciliogenesis in the pronephros and within the Kupffer's vesicle and results in aberrant L-R axis determination. Our results highlight a critical role for Ccdc11 in the assembly and function of motile cilia and implicate centriolar satellite-associated proteins as a new class of proteins in the pathology of L-R patterning and congenital heart disease.

A double-hotdog with a new trick: structure and mechanism of the trans-acyltransferase polyketide synthase enoyl-isomerase.

Many polyketide natural products exhibit invaluable medicinal properties, yet much remains to be understood regarding the machinery responsible for their biosynthesis. The recently discovered trans-acyltransferase polyketide synthases employ processing enzymes that catalyze modifications unique from those of the classical cis-acyltransferase polyketide synthases. The enoyl-isomerase domains of these megasynthases shift double bonds and are well-represented by an enzyme that helps forge the triene system within the antibiotic produced by the prototypical bacillaene synthase. This first crystal structure of an enoyl-isomerase, at 1.73 Å resolution, not only revealed relationships between this class of enzymes and dehydratases but also guided an investigation into the mechanism of double bond migration. The catalytic histidine, positioned differently from that of dehydratases, was demonstrated to independently shuttle a proton between the γ- and α-positions of the intermediate. This unprecedented mechanism highlights the catalytic diversity of divergent enzymes within trans-acyltransferase polyketide synthases.

ZMYND10 is mutated in primary ciliary dyskinesia and interacts with LRRC6.

Defects of motile cilia cause primary ciliary dyskinesia (PCD), characterized by recurrent respiratory infections and male infertility. Using whole-exome resequencing and high-throughput mutation analysis, we identified recessive biallelic mutations in ZMYND10 in 14 families and mutations in the recently identified LRRC6 in 13 families. We show that ZMYND10 and LRRC6 interact and that certain ZMYND10 and LRRC6 mutations abrogate the interaction between the LRRC6 CS domain and the ZMYND10 C-terminal domain. Additionally, ZMYND10 and LRRC6 colocalize with the centriole markers SAS6 and PCM1. Mutations in ZMYND10 result in the absence of the axonemal protein components DNAH5 and DNALI1 from respiratory cilia. Animal models support the association between ZMYND10 and human PCD, given that zmynd10 knockdown in zebrafish caused ciliary paralysis leading to cystic kidneys and otolith defects and that knockdown in Xenopus interfered with ciliogenesis. Our findings suggest that a cytoplasmic protein complex containing ZMYND10 and LRRC6 is necessary for motile ciliary function.

Evidence for metabolic imbalance of vitamin A2 in wild fish chronically exposed to metals.

In a recent study on indigenous yellow perch chronically exposed to metals, we reported a negative correlation between liver metal concentration and liver transcription levels of genes encoding for enzymes involved in the metabolism of retinoids. We therefore speculated that metals, and especially the non-essential metal Cd, could alter the metabolism of retinoids in wild fish. Thus the present field study investigates the impact of in situ metal exposure on retinoid storage. A total of 55 yellow perch (Perca flavescens) were sampled in six lakes representing a metal contamination gradient (8≤N≤10 per lake). Our results show that yellow perch from Cd-contaminated lakes had significantly higher concentrations of liver dehydroretinol and dehydroretinyl esters than did fish from reference lakes. However, the increase in retinyl ester stores with increasing Cd concentrations was quantitatively much more important than the increase in free dehydroretinol. As a result, a significant decrease in the percentage of hepatic free dehydroretinol with increasing renal Cd concentrations was observed. These results suggest that the enzymes and the binding proteins involved in vitamin A homeostasis are inhibited by the presence of Cd. Alternatively, the increase in tissue vitamin A (antioxidant) levels could serve to better counteract the oxidative stress engendered by Cd exposure. Overall our findings illustrate that vitamin A(2) homeostasis can be altered as a consequence of chronic exposure to low Cd concentrations. Thus, in the context of environmental risk assessment, the percentage of liver free dehydroretinol can be considered as a biomarker of for in situ Cd exposure.

Apical movement during interkinetic nuclear migration is a two-step process.

Neural progenitor cells in the pseudostratified neuroepithelium in vertebrates undergo interkinetic nuclear migration, which results in mitotic cells localized to the apical surface. Interphase nuclei are distributed throughout the rest of the epithelium. How mitosis is coordinated with nuclear movement is unknown, and the mechanism by which the nucleus migrates apically is controversial. Using time-lapse confocal microscopy, we show that nuclei migrate apically in G2 phase via microtubules. However, late in G2, centrosomes leave the apical surface after cilia are disassembled, and mitosis initiates away from the apical surface. The mitotic cell then rounds up to the apical surface, which is an actin-dependent process. This behavior is observed in both chicken neural-tube-slice preparations and in mouse cortical slices, and therefore is likely to be a general feature of interkinetic nuclear migration. We propose a new model for interkinetic nuclear migration in which actin and microtubules are used to position the mitotic cell at the apical surface.

Oxidation of retinoic acids in hepatic microsomes of wild bullfrogs Lithobates catesbeianus environmentally-exposed to a gradient of agricultural contamination.

Agricultural contaminants are suspected of contributing to the increased incidence of deformities and the decline of amphibians populations worldwide. Many authors have further suggested that a retinoid effect could be implicated in teratogenic mechanisms since the reported deformities resemble those caused by abnormal levels of retinoic acid (RA). We previously reported altered retinoid concentrations in male bullfrogs from the Yamaska River basin (Québec, Canada) associated with moderate-to-high agricultural activity, and the findings were consistent with a possible effect on hepatic RA oxidation. An in vitro assay was therefore optimized and hepatic microsomal RA oxidation in bullfrogs was found to be quite different from that of other vertebrates. With either all-transRA (atRA) or 13cisRA as the substrate, the major metabolite generated was at4-oxo-RA. The reaction with 13cisRA as substrate, markedly greater compared with atRA, was enhanced in the presence of a reducing agent and inhibited by cytochrome P450 inhibitors in a dose-dependent manner. Hepatic RA oxidation in male bullfrogs showed significant differences between sites with no clear relationship to a gradient of agricultural activity or 13cis-4-oxo-RA quantified in plasma. In contrast, the in vitro RA oxidation in females increased with the levels of contamination and coincided in vivo with higher plasma 13cis-4-oxo-RA concentration. The levels of circulating 4-oxo-derivatives could be influenced by hepatic RA oxidative metabolism as well as isomerization conditions or RA precursor levels.

Interkinetic nuclear migration: a mysterious process in search of a function.

During interkinetic nuclear migration (INM), the nuclei in many epithelial cells migrate between the apical and basal surfaces, coordinating with the cell cycle, and undergoing cytokinesis at the apical surface. INM is observed in a wide variety of tissues and species. Recent advances in time-lapse microscopy have provided clues about the mechanisms and functions of INM. Whether actin or microtubules are responsible for nuclear migration is controversial. How mitosis is initiated during INM is poorly understood, as is the relationship between the cell cycle and nuclear movement. It is possible that the disagreements stem from differences in the tissues being studied, since epithelia undergoing INM vary greatly in terms of cell height and cell fates. In this review we examine the reports addressing the mode and mechanisms that regulate INM and suggest possible functions for this dramatic event.

Changes of thyroid hormone levels and related gene expression in Chinese rare minnow (Gobiocypris rarus) during 3-amino-1,2,4-triazole exposure and recovery.

Thyroid hormones (THs) play an important role in the development and metabolism of fish through their influences on genetic transcription and are targets for endocrine disruptive agents in the aquatic environment. Amitrole is a pesticide potentially interfering with thyroid hormone regulation. In this study, the rare minnow (Gobiocypris rarus) was exposed to different levels of 3-amino-1,2,4-triazole (amitrole) and allowed to recover in clean water. Plasma TH levels and the expression of TH-related genes, including transthyretin (ttr), deiodinases (d1 and d2), and the thyroid hormone receptor (tralpha) from the livers and brains were evaluated. After exposure, the plasma TH levels did not change. Histopathological observations showed that livers were degenerated at 10,000 ng/l and these damages could be recovered by the withdrawal of amitrole. However, the ttr, d1, and d2 mRNA levels in the livers of males were significantly up-regulated in all exposure groups (p<0.05). The ttr and d2 mRNA levels were significantly up-regulated at 10,000 ng/l and 10, 100, and 1000 ng/l in the livers of females, respectively (p<0.05). In the brains of males, a twofold increase of d2 mRNA levels at > or = 100 ng/l and a fivefold decrease of tralpha mRNA levels at > or = 10 ng/l were observed (p<0.05), whereas no significant differences were observed in the expression of d2 and tralpha in the brains of females. After a recovery period, the ttr, d1, and d2 mRNA levels in the livers of males returned to control levels, but the tralpha mRNA levels were irreversibly decreased at all treatments (p<0.05). In addition, the d2 mRNA levels in the livers of females were significantly induced at > or = 100 ng/l. Moreover, the d2 mRNA levels in the brains of males and females were up-regulated at 10,000 ng/l. These results indicated that amitrole exposure could result in alternations of ttr, d1, d2, and tralpha gene expression in different tissues of the rare minnow. The expression of these TH-related genes in males was more sensitive to amitrole than those of females. Recovery in clean water was associated with the selective regulation of TH-related gene transcription in the rare minnow. Therefore, these TH-related genes can serve as biomarkers to screen the effects of thyroid disruption chemicals in rare minnow.

Combined effects of agricultural activity and parasites on biomarkers in the bullfrog, Rana catasbeiana.

Agricultural contaminants can have devastating impacts on amphibian survival and development, particularly considering their sensitivity to environmental perturbation. However, it is commonly overlooked that amphibians are infected with various parasites that can influence the overall health of the animal when exposed to a stressful environment. We investigated the interaction of agriculture and parasitism on the health of bullfrogs (Rana catesbeiana) in the field. Nine physiological and immunological biomarkers were related to naturally acquired parasite infections, along a gradient of agricultural activity. Most health biomarkers were affected by agriculture, parasitism, or both. Although bullfrogs residing in agricultural areas were infected with fewer parasite species, reflecting environmentally compromised ecosystems, certain persistent parasites interacted with agricultural disturbance to alter the physiology and immune competence of bullfrogs. The consequences of the combination for animal health highlight the importance of parasitism in ecotoxicological studies. Consideration of parasitism is warranted when evaluating the influence of anthropogenic disturbance on amphibian declines and environmental health.

Study design, water quality, morphometrics and age of the bullfrog, Rana catesbeiana, in sub-watersheds of the Yamaska River drainage basin, Québec, Canada.

The Yamaska River basin is renowned for its poor water quality, which has been attributed to intensive agriculture (corn, soya, high-density pork and poultry production). Six locations within the Yamaska watershed were selected to evaluate the impact of agriculture on water habitats and study the bullfrog as a sentinel species of potential exposure and effects. The selected sub-watersheds were chosen according to the percentage of surface area under cultivation and classified as low (0-19%), moderate (20-59%) or high (>60%). In 2004 and 2005, analysis of surface water samples demonstrated that pesticide concentrations and most water quality parameters increased with increasing agricultural activity. Sixteen adult bullfrogs were sampled from each site. Animals were weighed, measured for length (total, snout-vent, tibia, tympanum), sexed, and evaluated for the colour of the throat. Skeletochronology was used to estimate the growth and age of the frogs. Cross-sections of decalcified phalanges and femurs were treated in order to count LAGs (lines of arrest growth) and for the observations of other parameters related to bone growth. The bullfrogs from highly contaminated sites had the lowest mean age and the smallest snout-vent length compared to sites of low contamination.

Occurrence and distribution of organochlorine pesticides - lindane, p,p'-DDT, and heptachlor epoxide - in surface water of China.

Persistent organochlorine pesticides pollutants (OCPs) have been reported to occur at relatively high concentrations in some Chinese waters. In order to map the distribution of organochlorine pesticides in the surface water throughout China, samples were collected from over 600 sites in seven major river basins and three main internal rivers drainage areas during 2003 and 2004. The surface water samples were analyzed for the representative organochlorine pesticides contaminants including lindane (gamma-HCH), p,p'-DDT and heptachlor epoxide. In general, the most frequently detected compound was lindane, being detected in 83.9% of samples (mean=31.3 ng/l; range <0.17-860 ng/l), and the highest concentration was present in the Yellow River basin. p,p'-DDT was detected in 63.1% of the samples collected (mean=14.6 ng/l; range <0.14-368 ng/l) with the highest concentration present in the Huaihe River basin. Heptachlor epoxide was detected in only 9.3% of water samples (range <0.11-10 ng/l). Measured concentrations for the three compounds were low and rarely exceed the environment quality standard for surface water of China. Lindane was more frequently detected at much higher concentrations in the rivers of northern China compared with those of southern China. The sites with higher concentration of lindane and p,p'-DDT mainly occurred in the Yellow River and Huaihe River basins, so the results of this investigation indicate that the organochlorine pesticide contamination of Yellow River and Huaihe River basins should be of particular concern relative to the other basins. When compared with other regions of the world, it appears that the Chinese surface water is moderately polluted by lindane and p,p'-DDT.

Gender differences in the effects of organochlorines, mercury, and lead on thyroid hormone levels in lakeside communities of Quebec (Canada).

Environmental chemicals can disrupt endocrine balance and in particular thyroid hormone (TH) homeostasis. However, studies differ with respect to thyroid profile changes and gender differences are rarely examined. This study investigated the THs, triodothyronine (T3), thyroxine (T4), and thyroid stimulating hormone (TSH), in relation to serum organochlorines (OCs), bioindicators of mercury (Hg) and blood lead (Pb) in 211 freshwater fish consumers (124 men and 87 women) from two communities in Canada. Thyroid hormones were within the normal range and the bioindicators of exposure were low compared to other reports on fish consumers. Stratified analysis showed that for women, serum T3 concentrations were negatively related to serum concentrations of PCB 138, PCB 153, the non-coplanar congeners, Arochlor 1260, and SigmaPCB, as well as p,p'-DDE. No relations were observed between T4 and any of the chemicals measured, but TSH was negatively related to blood Pb. For men, serum T4 was inversely related to PCB 138, non-ortho-substituted (dioxin-like) PCBs and SigmaPCB. A significant positive relationship was observed between serum TSH and different PCB congeners (PCB 138, PCB 180, non-coplanar congeners, mono-ortho coplanar congeners, dioxin-like PCBs), as well as SigmaPCB. Serum TSH increased with hair and blood Hg concentrations and was highest among those in the highest 50th percentile for both Hg and dioxin-like PCB congeners compared to the others. No associations were observed for T3 in men. These findings suggest that even at low concentrations, these environmental contaminants can interfere with thyroid status and effects may differ by gender.

Comparison of ethinylestradiol and nonylphenol effects on reproduction of Chinese rare minnows (Gobiocypris rarus).

Breeding pairs of rare minnows were exposed to 5, 10, 20 microg/L 4-nonylphenol (NP) or 4 ng/L 17alpha-ethinylestradiol (EE(2)) for 21 days. Among reproductive parameters investigated, fecundity, fertility, and laying interval were significantly affected by 4 ng/L EE(2), but were not affected at any exposure level of NP. These effects coincided with male feminization, ova-testis, increased plasma VTG in both males and females as well as the accumulation of eosinophilic material in kidneys and livers. Histopathologic evaluation revealed severe kidney lesions in both EE(2) and NP-exposed fish. The bioassay succeeded in demonstrating that EE(2) was several orders of magnitude more potent than the industrial chemical NP. The rare minnow was more sensitive to EE(2) than medaka, but comparable to zebrafish in terms of sensitivity. The results of the present investigation indicated that a short-term reproductive bioassay is feasible using the Chinese rare minnow as the test species.

Assessment of 17alpha-ethinylestradiol effects and underlying mechanisms in a continuous, multigeneration exposure of the Chinese rare minnow (Gobiocypris rarus).

17alpha-Ethinylestradiol (EE(2)) is a synthetic estrogen used primarily in birth control pills and in hormone replacement therapy. Owing to its occurrence in surface waters at concentrations frequently greater than 1 ng/l and its projected future use, EE(2) is expected to pose a significant risk to aquatic organisms. This study was conducted to obtain long-term exposure data necessary for the establishment of water quality criteria and to investigate mechanisms associated with toxic effects. In a multigeneration experiment, Chinese rare minnows (Gobiocypris rarus) were constantly exposed to environmentally relevant concentrations of the synthetic estrogen EE(2). Mortality, deformities, reproductive parameters, plasma vitellogenin and histopathology were assessed. The results showed that, in the F(0) generation, all endpoints were significantly affected at concentrations higher than 0.2 ng/l EE(2). No F(1) phenotypic males developed to maturity at 0.2 ng/l and, when adult females of this exposure group were crossed with unexposed males, no F(2) fertile eggs were produced. Kidney histopathology and ultrastructure suggest anomalies possibly associated with increased vitellogenin accumulation. We concluded that the reproduction of the F(1) minnows was completely inhibited at the lowest concentration tested, 0.2 ng/l EE(2), a concentration frequently detected in surface waters. Growth effects may be related to increased energy requirements including the energy used in VTG synthesis. Reproductive effects are presumably associated with male feminization and the occurrence of testis-ova in males; however, ovarian degeneration observed in females may also have contributed to reproductive failure.

Predicting bioavailability and accumulation of organochlorine pesticides by Japanese medaka in the presence of humic acid and natural organic matter using passive sampling membranes.

Adsorption to dissolved organic matter (DOM) may significantly decrease the freely dissolved concentration of many hydrophobic organic compounds and, hence, result in reduced bioavailability to aquatic organisms. Here, the suitability of using triolein-embedded cellulose acetate membrane (TECAM) as a biomimetic surrogate to assess the bioavailability of organochlorine pesticides (OCPs) in water in the presence of DOM was explored. The accumulation of OCPs was measured in TECAM and pelagic Japanese medaka (Oryzias latipes) in the laboratory after 12 h exposure to water containing different levels of Aldrich humic acid. Further, OCP uptake by TECAM and medaka in real aqueous environments was evaluated after 30 d exposures in two sites. Laboratory results showed that OCP uptake by medaka consistently decreased with increasing levels of humic acid in the range of 0-15 mg C/L in sample solutions. This tendency was closely mimicked by OCP accumulation in TECAM under the same conditions. Field results showed that TECAM accumulated similar OCP patterns as medaka (r2 = 0.92 for site 1 and r2 = 0.94 for site 2), although comparison of the in-field eight OCP concentrations in TECAM to those in medaka yielded approximately a factor of 3 (on a wet weight basis). These results suggest that the TECAM method can be used as a simple and useful tool to predict the bioavailability and bioaccumulation potential of poorly biotransformed organic compounds in pelagic fish in aqueous environment.

Effects of HMX-lead mixtures on reproduction of the earthworm Eisenia andrei.

High metal (e.g., Pb) concentrations are typically found in explosive-contaminated soil, and their presence may increase, decrease, or not influence toxicity predicted on the basis of one explosive alone (e.g., HMX). Nevertheless, few data are available in the scientific literature for this type of multiple exposure. Soil organisms, such as earthworms, are one of the first receptors affected by the contamination of soil. Therefore, a reproductive study was conducted using Eisenia andrei in a forest-type soil. Both HMX and Pb decreased reproduction parameters (number of total cocoons, hatched cocoons, and surviving juveniles) individually. Based on the total number of cocoons, HMX was more toxic in a forest soil than Pb, with EC(50) of 31 mg kg(-1), and 1068 mg kg(-1), respectively. The slope of the concentration-response curve was significantly greater in the case of Pb, which is consistent with the possibility that the two compounds do not act on the same target site. The response-addition model was used to predict the response of earthworms and to test for interaction between the two contaminants. The predicted toxicity was not significantly different than the observed toxicity, implying that Pb and HMX were considered noninteractive compounds. The combined action of Pb-HMX may be described, therefore, as dissimilar-noninteractive joint action in a forest soil. The results illustrate the relevance of considering the presence of metals in the risk assessment of explosive-contaminated sites because metals can add their toxicity to explosives. Extension of this study to other types of soil and other metals would improve the understanding of toxicity at these sites.

Toxicity of the aromatase inhibitor letrozole to Japanese medaka (Oryzias latipes) eggs, larvae and breeding adults.

Letrozole is a synthetic aromatase inhibitor and interferes in the committed step in the synthesis of endogenous estrogens from androgens. To evaluate potential effects on the early life stages of Japanese medaka, larvae and fertilized eggs were exposed to letrozole for 96 h and 14 days, respectively. No larvae died and no adverse effects were found on embryonic development at concentrations up to 3125 microg/L. Reproductive effects were assessed by exposing adults to 1, 5, 25, 125 and 625 microg/L letrozole for 21 days. A dose-dependent decrease in fecundity (>25 microg/L) and fertility (>5 microg/L) accompanied by histological changes suggested the inhibition of oocyte growth and possibly maturation. At 625 microg/L, the fish ceased spawning during the last week of exposure. Letrozole (>5 microg/L) reduced plasma vitellogenin levels in females in a dose-dependent manner. Transgenerational effects were evaluated by removing freshly-laid F1 eggs from letrozole-contaminated water and raising them to 15 days post-hatching in control water. Hatchability and time to hatching were detrimentally affected (>5 microg/L), but no morphological deformities were observed. Furthermore, a dose-dependent increase in the proportion of genotypic F1 males was found (>5 microg/L).

Tamoxifen effects on the early life stages and reproduction of Japanese medaka (Oryzias latipes).

Tamoxifen is widely used in the treatment of breast cancer and can enter the aquatic environment in municipal wastewater. To evaluate potential effects on embryonic development of Japanese medaka, fertilized eggs were exposed to tamoxifen at 1-625µg/l for 14 days. Adverse effects on hatchability and time to hatching only occurred at 125 and 625µg/l. Reproductive effects were assessed by exposing adults for 21 days to the same dose range. At all concentrations tested, tamoxifen significantly increased plasma vitellogenin levels in males in a dose-dependent manner. Fecundity and fertility were detrimentally affected at 625µg/l. Additionally, F1 eggs were removed from tamoxifen-contaminated water to evaluate transgenerational effects. Hatchability was affected at 625µg/l but no morphological deformities were observed. A significant dose-dependent increase in the proportion of genotypic males occurred at all concentrations greater than 5µg/l.