Progestagenic compounds are emerging contaminants found in surface and ground water around the world. Information on the effects and potency of progestagens is needed in order to understand the environmental risks posed by these compounds. Using the Xenopus (Silurana) tropicalis test system, developmental and reproductive toxicity after exposure to selected progestagens were determined. Larval exposure to levonorgestrel (LNG) severely impaired oviduct and ovary development causing sterility. No effects on testicular development, spermcount or male fertility were observed. Hepatic mRNA expression of the androgen receptor was increased in the females indicating that the receptor is involved in LNG-induced developmental reproductive toxicity. Exposure of adult females to LNG, norethindrone (NET) or progesterone (P) increased the proportions of previtellogenic oocytes and reduced the proportions of vitellogenic oocytes compared with the controls, indicating an inhibited vitellogenesis. The effects on oocyte development were ascertained at environmentally relevant concentrations of LNG, NET and P (1.3, 1 and 10 ng/L respectively). Since unintentional co-exposure of progestagens and ethinylestradiol (EE2) occurs in wildlife and also in human infants, data on mixture effects of combined exposures to these hormones during development are needed. Co-exposure during development showed antagonistic effects of EE2 and LNG. EE2 caused a female biased sex ratio which showed a tendency to be antagonized by LNG. Moreover, the hepatic AR induction by LNG was counteracted by co-exposure to EE2. In conclusion, the results show that female amphibians are susceptible to reproductive toxicity of progestagens after developmental exposure as well as after adult exposure during the breeding period. The differentiating Müllerianduct and ovary, and the egg development are sensitive targets for progestagens. Finally, the findings reported in this thesis show that environmental progestagens impairs reproductive function in amphibians and may present a threat to reproduction in wild populations.

Orientador: Alexandre Ninhaus Silveira / Resumo: Pseudoplatystoma reticulatum, Leiarius marmoratus e o híbrido resultante do cruzamento entre essas espécies apresentam características zootécnicas interessantes e promissoras para o cultivo em piscicultura. Porém, a larvicultura dessas espécies apresentam altos índices de mortalidade, pois nesse período as larvas não estão totalmente desenvolvidas e não possuem o sistema digestório completamente formado. Pesquisas relacionadas ao desenvolvimento do sistema digestório de larvas tem sido uma ferramenta importante para o conhecimento sobre os períodos de alimentação endógena, exógena e mista. Neste sentido, este trabalho teve como objetivo analisar e comparar as características anatômicas, histológicas e histoquímicas do sistema digestório de Pseudoplatystoma reticulatum, Leiarius marmoratus e do híbrido, nas fases inicias de desenvolvimento após eclosão. As amostras foram fixadas em solução de karnovsky (paraformoldeído 4% + glutaraldeído 2% + tampão fosfato Sorensen 0,1 M, pH 7,2), e processadas para análise anatômica, histológica e histoquímica. As larvas de P. reticulatum, L. marmoratus e híbrido eclodiram 24 horas após a fertilização. As larvas recém-eclodidas apresentaram-se pouco desenvolvidas e com escassez de cromatóforos. A cavidade bucofaríngea pôde ser observada, mas não foi possível diferenciar os componentes do tubo digestivo. As reservas de vitelo foram observadas até o 3º DPE em P. reticulatum e em L. marmoratus e no Híbrido até o 4º DPE. Para os três modelos bio... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Pseudoplatystoma reticulatum, Leiarius marmoratus and the hybrid resulting from the cross between these species show interesting and promising zootechnical characteristics for fish farming. The main bottleneck of larviculture today is the high mortality rate in the early stages of life, because in this period the larvae are not fully developed and do not have a fully formed digestive system. Research related to the development of the digestive system of larvae, has been an important tool for understanding the periods of endogenous, exogenous and mixed feeding. In this sense, this work aimed to analyze and compare the anatomical, histological and histochemical characteristics of the digestive system of Pseudoplatystoma reticulatum, Leiarius marmoratus and the hybrid, in the early stages of development after hatching. The samples were fixed in a karnovsky solution (paraformoldehyde 4% + glutaraldehyde 2% + phosphate buffer Sorensen 0.1 M, pH 7.2), and processed for anatomical, histological and histochemical analysis. The larvae of P. reticulatum, L. marmoratus and hybrid hatched 24 hours after fertilization. The newly hatched larvae were poorly developed, with a shortage of chromatophores. The buccopharyngeal cavity could be observed, but it was not possible to differentiate the components of the digestive tract. Calf reserves were observed until the 3º DPE in P. reticulatum and in L. marmoratus and in the hybrid until the 4º DPE. For the three biological models, the first gobl... (Complete abstract click electronic access below) / Mestre

<p>A number of environmental pollutants have been shown to mimick the action of the female sex hormone estrogen and are, therefore, suspected to be responsible for reproductive abnormalities seen in wildlife. Test systems which can be used in hazard and risk assessment of chemicals with estrogenic effects are consequently needed. In this thesis, I propose the avian egg as an <i>in vivo</i> test system for estrogenic compounds. I conclude that malformation of the left testis and the Müllerian ducts (MDs: embryonic oviducts) in avian embryos can be used as endpoints to examine estrogenic activity of chemicals. MD malformation is more easily determined and thereby faster to use as an endpoint than histologically observed feminization of the testis. The usefulness of MD/oviduct malformations as biomarkers for estrogenic effects in wild birds should be considered. </p><p>The environmental pollutants bisphenol A (BPA) and <i>o,p´</i>-DDT induced similar effects as the synthetic estrogens, ethynylestradiol and diethylstilbestrol. BPA caused MD malformations in quail embryos and ovotestis formation in chicken embryos. <i>o,p´</i>-DDT induced MD malformations in both quail and chicken embryos and ovotestis in chicken embryos. The flame retardant, tetrabromobisphenol A did not induce estrogen-like effects in quail or chicken embryos, but showed a relatively high embryolethality. </p><p>Embryonic exposure to estrogen caused persisting malformations of the oviduct, as well as a changed distribution pattern of the enzyme carbonic anhydrase in the shell gland of adult females. Considering the crucial role of carbonic anhydrase in shell formation, such changes could result in decreased shell quality. I propose that eggshell thinning in avian wildlife could reflect a functional malformation in the shell gland that is induced by xeno-estrogens during embryonic development, rather than being caused by exposure of the adult bird to environmental pollutants. This hypothesis opens new possibilities for studying the mechanisms behind contaminant-induced eggshell thinning in birds.</p>

A number of environmental pollutants have been shown to mimick the action of the female sex hormone estrogen and are, therefore, suspected to be responsible for reproductive abnormalities seen in wildlife. Test systems which can be used in hazard and risk assessment of chemicals with estrogenic effects are consequently needed. In this thesis, I propose the avian egg as an in vivo test system for estrogenic compounds. I conclude that malformation of the left testis and the Müllerian ducts (MDs: embryonic oviducts) in avian embryos can be used as endpoints to examine estrogenic activity of chemicals. MD malformation is more easily determined and thereby faster to use as an endpoint than histologically observed feminization of the testis. The usefulness of MD/oviduct malformations as biomarkers for estrogenic effects in wild birds should be considered. The environmental pollutants bisphenol A (BPA) and o,p´-DDT induced similar effects as the synthetic estrogens, ethynylestradiol and diethylstilbestrol. BPA caused MD malformations in quail embryos and ovotestis formation in chicken embryos. o,p´-DDT induced MD malformations in both quail and chicken embryos and ovotestis in chicken embryos. The flame retardant, tetrabromobisphenol A did not induce estrogen-like effects in quail or chicken embryos, but showed a relatively high embryolethality. Embryonic exposure to estrogen caused persisting malformations of the oviduct, as well as a changed distribution pattern of the enzyme carbonic anhydrase in the shell gland of adult females. Considering the crucial role of carbonic anhydrase in shell formation, such changes could result in decreased shell quality. I propose that eggshell thinning in avian wildlife could reflect a functional malformation in the shell gland that is induced by xeno-estrogens during embryonic development, rather than being caused by exposure of the adult bird to environmental pollutants. This hypothesis opens new possibilities for studying the mechanisms behind contaminant-induced eggshell thinning in birds.