This work explores the metabolism of mercury in liver and spleen tissue of fish from a methylmercury contaminated wetland. Wild-caught bass, catfish, bowfin and gar were collected. Macrophage centers, which are both reactive and primary germinal centers in various fish tissues, were hypothesized to be the cause of demethylation of methylmercury in fish tissue. Macrophage centers are differentially expressed in fish tissue based on phylogenetic lineage, and are found primarily in the livers of preteleostean fish and in the spleen of teleostean fish. Histology of liver and spleen was examined in both control and wild-caught fish for pathology, size and number of macrophage centers, and for localization of mercury. Total mercury was estimated in the muscle tissue of all fish by direct mercury analysis. Selenium and mercury concentrations were examined in the livers of wild-caught fish by liquid introduction inductively coupled plasma mass spectrometry (ICP-MS). Total mercury was localized in histologic sections by laser ablation ICP-MS (LA-ICP-MS). Mercury speciation was determined for inorganic and methylmercury in liver and spleen of fish by bas chromatography-cold vapor atomic fluorescence spectroscopy (GC-CVAFS). Macrophage center tissue distribution was found to be consistent with the literature, with a predominance of centers in preteleostean liver and in spleens of teleostean fish. Little evidence histopathology was found in the livers or spleens of fish examined, but differences in morphology of macrophage centers and liver tissue across species are noted. the sole sign of liver pathology noted was increased hepatic hemosiderosis in fish with high proportions of liver inorganic mercury. Inorganic mercury was found to predominate in the livers of all fish but bass. Organic mercury was found to predominate in the spleens of all fish. Mercury was found to accumulate in macrophage centers, but concentrations of mercury in this compartment were found to vary less in relation to total mercury than hepatocyte mercury. No association was found between selenium content and inorganic mercury proportions. Overall, findings from this study to not support a primary role for macrophage centers in the demethylation of methylmercury in fish tissues.

Made available in DSpace on 2014-06-11T19:30:55Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-06-01Bitstream added on 2014-06-13T19:01:01Z : No. of bitstreams: 1 zieri_r_dr_sjrp.pdf: 889017 bytes, checksum: 170c2d62d9c91d29ca0d8ba71a162b3b (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Os vertebrados ectotérmicos apresentam um desenvolvido sistema de células com pigmentos melânicos em seu citoplasma, localizado em diversos órgãos e membranas. A dispersão dos grãos de melanina dentro dos melanócitos responde fortemente à exposição ao hormônio estimulante de melanócitos (MSH) e aos hormônios sexuais, promovendo alteração na coloração. Este trabalho buscou avaliar o efeito dos esteróides cipinato de testosterona, 17 β-estradiol e do MSH sobre a pigmentação hepática e testicular de Eupemphix nattereri, mediante análise histológica, e ultra-estrutural. Foram coletados 60 machos adultos na região de São José do Rio Preto, SP. Para cada experimento, dez machos adultos receberam doses subcutâneas contendo 0,5 mg/kg de cipionato de testosterona e 0,5 mg/kg de 17 β-estradiol, dissolvidos em óleo vegetal e, 2,5x10-7 mmol/10g de α-MSH dissolvido em PBS, durante 7 dias, a cada 48h. O grupo controle recebeu apenas óleo vegetal ou PBS. Grupos com cinco animais foram eutanasiados 24h após do término do tratamento, e outros cinco, após 15 dias. Foram realizados estudos morfológicos utilizando microscopia de luz e microscopia eletrônica de transmissão. No tecido hepático foram encontradas células de Kupffer. No interstício testicular observou-se melanócitos, com inúmeros melanossomos. Para o grupo tratado com cipionato de testosterona, foi observado um aumento de aproximadamente 2x na área ocupada pelas células pigmentadas no fígado e 4x nos testículos mediante analise a curto prazo. Para o mesmo tratamento, foi observado aumento de 1,8x no fígado dos animais eutanasiados após 15 dias. Tal resposta pode ser decorrente do aumento da atividade melanossintética, do aumento do número de células, ou ambos. Os animais tratados com 17 β-estradiol e α- MSH não apresentaram alterações na pigmentação hepática e testicular mediante... / Ectothermic vertebrates have a well-developed system of melanin-containing cells, which are distributed throughout several organs and tissues. The dispersion of melanin-containing granules within melanocytes responds to prolonged exposure to high concentration of MSH and sexual hormones inducing color change. The present study aimed to characterize morphological and stereological patterns of pigmented cells in liver and testis of the Eupemphix nattereri under effect of testosterone cypionate, 17 β-estradiol and α-MSH. Sixty adult males were collected in São José do Rio Preto, São Paulo State, Brazil. For each experiment, ten adult males were treated with the following hormones injected in the dorsal lymph sac during seven days: (1) 5 mg/kg dose of testosterone cypionate or (2) 5 mg/kg dose of 17 β-estradiol (dissolved in vegetal oil) or (3) 2,5x10-7 mmol/10g of α-MSH (dissolved in PBS). The control group received only vegetal oil or PBS solution at the same hormonal concentration. Groups of five animals were euthanatized after 24h and the other five, after 15 days. Morphological studies with light and transmission electron microscopy were carried out in the groups. In the hepatic tissue were found Kupffer cells, which are characterized by multivesicle bodies in the cytoplasm and large amount of melanosomes. In the testis, melanocytes-like cells are present in the interstitium. They presented large and irregular aspect and a great amount of intensely pigmented cytoplasm. For the groups treated with testosterone cypionate an increase of approximately 2x in the occupied area by the pigmented cells in the liver and 4x in the testis was observed in the treatment group against the control. The observed response can be due to increase of melanosynthesis Introdução 3 Zieri et al., 2010 activity, increase in the number of pigmented cells, or both. Moreover, for the same treatment... (Complete abstract click electronic access below)

Resumo: Os vertebrados ectotérmicos apresentam um desenvolvido sistema de células com pigmentos melânicos em seu citoplasma, localizado em diversos órgãos e membranas. A dispersão dos grãos de melanina dentro dos melanócitos responde fortemente à exposição ao hormônio estimulante de melanócitos (MSH) e aos hormônios sexuais, promovendo alteração na coloração. Este trabalho buscou avaliar o efeito dos esteróides cipinato de testosterona, 17 β-estradiol e do MSH sobre a pigmentação hepática e testicular de Eupemphix nattereri, mediante análise histológica, e ultra-estrutural. Foram coletados 60 machos adultos na região de São José do Rio Preto, SP. Para cada experimento, dez machos adultos receberam doses subcutâneas contendo 0,5 mg/kg de cipionato de testosterona e 0,5 mg/kg de 17 β-estradiol, dissolvidos em óleo vegetal e, 2,5x10-7 mmol/10g de α-MSH dissolvido em PBS, durante 7 dias, a cada 48h. O grupo controle recebeu apenas óleo vegetal ou PBS. Grupos com cinco animais foram eutanasiados 24h após do término do tratamento, e outros cinco, após 15 dias. Foram realizados estudos morfológicos utilizando microscopia de luz e microscopia eletrônica de transmissão. No tecido hepático foram encontradas células de Kupffer. No interstício testicular observou-se melanócitos, com inúmeros melanossomos. Para o grupo tratado com cipionato de testosterona, foi observado um aumento de aproximadamente 2x na área ocupada pelas células pigmentadas no fígado e 4x nos testículos mediante analise a curto prazo. Para o mesmo tratamento, foi observado aumento de 1,8x no fígado dos animais eutanasiados após 15 dias. Tal resposta pode ser decorrente do aumento da atividade melanossintética, do aumento do número de células, ou ambos. Os animais tratados com 17 β-estradiol e α- MSH não apresentaram alterações na pigmentação hepática e testicular mediante... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Ectothermic vertebrates have a well-developed system of melanin-containing cells, which are distributed throughout several organs and tissues. The dispersion of melanin-containing granules within melanocytes responds to prolonged exposure to high concentration of MSH and sexual hormones inducing color change. The present study aimed to characterize morphological and stereological patterns of pigmented cells in liver and testis of the Eupemphix nattereri under effect of testosterone cypionate, 17 β-estradiol and α-MSH. Sixty adult males were collected in São José do Rio Preto, São Paulo State, Brazil. For each experiment, ten adult males were treated with the following hormones injected in the dorsal lymph sac during seven days: (1) 5 mg/kg dose of testosterone cypionate or (2) 5 mg/kg dose of 17 β-estradiol (dissolved in vegetal oil) or (3) 2,5x10-7 mmol/10g of α-MSH (dissolved in PBS). The control group received only vegetal oil or PBS solution at the same hormonal concentration. Groups of five animals were euthanatized after 24h and the other five, after 15 days. Morphological studies with light and transmission electron microscopy were carried out in the groups. In the hepatic tissue were found Kupffer cells, which are characterized by multivesicle bodies in the cytoplasm and large amount of melanosomes. In the testis, melanocytes-like cells are present in the interstitium. They presented large and irregular aspect and a great amount of intensely pigmented cytoplasm. For the groups treated with testosterone cypionate an increase of approximately 2x in the occupied area by the pigmented cells in the liver and 4x in the testis was observed in the treatment group against the control. The observed response can be due to increase of melanosynthesis Introdução 3 Zieri et al., 2010 activity, increase in the number of pigmented cells, or both. Moreover, for the same treatment... (Complete abstract click electronic access below) / Orientador: Classius de Oliveira / Coorientador: Sebastião Roberto Taboga / Banca: Rejane Maira Góes / Banca: Patrícia Simone Leite Vilamaior / Banca: Selma Maria Almeida Santos / Banca: Wagner José Fávaro / Doutor

Submitted by Liliane Ferreira (ljuvencia30@gmail.com) on 2018-06-27T14:01:23Z No. of bitstreams: 2 Dissertação - Gabriel Qualhato - 2018.pdf: 4517101 bytes, checksum: 7e23af60d803b2f275d7d9babce3cc73 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2018-06-28T10:51:30Z (GMT) No. of bitstreams: 2 Dissertação - Gabriel Qualhato - 2018.pdf: 4517101 bytes, checksum: 7e23af60d803b2f275d7d9babce3cc73 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2018-06-28T10:51:30Z (GMT). No. of bitstreams: 2 Dissertação - Gabriel Qualhato - 2018.pdf: 4517101 bytes, checksum: 7e23af60d803b2f275d7d9babce3cc73 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2018-02-28 / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / The environmental risk of nanomaterials (NMs) designed and used in nanoremediation process is of emerging concern, but their ecotoxilogical effects to aquatic organism remains unclear. In this present study, the citrate-coated (maghemite) nanoparticles (IONPs) were synthesized and its genotoxic, mutagenic, the melanomacrophage centers (MMC) response and hepatic histopathological biomarkers were investigated in female guppy. Fish were exposed to IONPs at environmentally relevant iron concentration (0.3 mg L-1) during 21 days and the animals were collected at the beginning of the experiment and after 3, 7, 14 and 21 days of exposure. The genotoxicity and mutagenicity were evaluated in terms of DNA damage (comet assay), micronucleus (MN) test, erythrocyte nuclear abnormalities (ENA) frequency and the liver were dissected and fixed dehydrated through increasing ethanol gradient, immersed in xylene PA, embedded in paraffin, performed of liver sections (5 μm thick) (3 sections per animal) in the microtome, stained by Mallory’s Trichrome and H. E. and analyzed. Results showed differential genotoxic and mutagenic effects of IONPs in the P. reticulata according to exposure time. The IONP induced DNA damage in P. reticulata after acute (3 and 7 days) and long-term exposure (14 and 21 days), while the mutagenic effects were observed only after long-term exposure. The DNA damage and the total ENA frequency increase linearly over the exposure time, indicating a higher induction rate of clastogenic and aneugenic effects in P. reticulata erythrocytes after long-term exposure to IONPs. Guppies exposed to IONPs showed increasing in the number, area and perimeter of MMC when compared to the unexposed ones, especially after 7 days of exposure. The results showed an increase in the frequency of histopathological changes in fish after the 7 days of exposure to IONPs, such micro- and macro-vesicular steatosis, melanomacrophage aggregates, exudate and haemorrhagic foci. The acute (3 and 7 days) and long-term (14 and 21 days) exposure of P. reticulata to IONPs induced high histopathological indexes associated with circulatory disorders and inflammatory responses. Results indicated that blood system and liver tissue of P. reticulata are excellent target organs to studies the ecotoxicity of IONPs. Confirming that P. reticulata is a biomonitor specie indicated for studies of ecotoxicity of IONPs. / O risco ambiental dos nanomateriais projetados e utilizados para nanoremediação se mostram hoje de grande interesse econômico e ambiental, mas seus efeitos ecotóxicológicos para os organismos aquáticos ainda permanecem obscuros. Neste presente estudo, as nanopartículas de oxido de ferro funcionalizadas com citrato (maghemita) (NOFs) foram sintetizadas e seus efeitos genotóxicos, mutagênicos, e histopatológicos foram investigados em fêmeas de Poecilia reticulata. Os peixes foram expostos a maghemita em concentrações de ferro ambientalmente relevantes (0,3 mg.L-1) durante 21 dias e os animais foram coletados no início do experimento e após 3, 7, 14 e 21 dias de exposição. A genotoxicidade e a mutagenicidade foram avaliadas por meio do dano causado ao DNA (Ensaio cometa), o teste do micronúcleo (MN) e anormalidades nucleares de eritrócitos (ANE) avaliaram o potencial mutagênico. Os fígados foram dissecados, fixados, desidratados em etanol, imersos em xilol, embebidos em parafina, e seccionados com 5 μm de espessura, os cortes corados em H.E. e Tricrômico de Mallory foram analisados e obtidos dados histomorfométricos. Os resultados mostraram efeitos genotóxicos e mutagênicos diferenciais das NOFs em P. reticulata de acordo com o tempo de exposição. As NOFs induziram danos ao DNA após exposição aguda (3 e 7 dias) e de longo prazo (14 e 21 dias), enquanto os efeitos mutagênicos foram observados somente para a exposição prolongada. O dano no DNA e a frequência total de ANE aumentaram linearmente ao longo do tempo de exposição, indicando uma maior taxa de indução de efeitos clastogênicos e aneugênicos em eritrócitos de P. reticulata após exposição prolongada a NOFs. Os guppies expostos também mostraram aumento do número, área e perímetro dos MMC quando comparados ao grupo controle, especialmente após 7 dias de exposição. Os resultados mostraram um aumento na frequência de alterações histopatológicas em peixes após os 7 dias de exposição as NOFs, tais como esteatose micro- e macro-vesicular, aumento dos MMC, exsudatos e focos hemorrágicos. A exposição aguda (3 e 7 dias) e a longo prazo (14 e 21 dias) de P. reticulata a NOFs induziu índices histopatológicos elevados associados a distúrbios circulatórios e respostas inflamatórias. Os resultados indicaram que os tecidos sanguíneo e hepático dos guppies são excelentes órgão alvo para estudar a ecotoxicidade das NOFs, confirmando que o P. reticulata é uma espécie biomonitora indicada para estudos sobre a ecotoxicidade das NOFs.

Orientador: Classius de Oliveira / Resumo: Peixes e anfíbios estão vulneráveis a diversos poluentes. O benzo[a]pireno (BaP) é um contaminante com propriedades tóxicas. Metabolizado pela enzima P450 (CYP1A1), de fase I EROD (Etoxiresorufina-O-deetilase), produz subprodutos tóxicos. Conhecida como um inibidor da CYP1A1, α-naftoflavona (aNF) impede o metabolismo do BaP e a geração de metabólitos tóxicos. A aNF é um composto exógeno e não se tem conhecimento do efeito dela em órgãos e tecidos. O fígado é a maior fonte de enzimas catalizadoras de reações de biotransformação. A suscetibilidade do órgão a danos por agentes químicos é uma conseqüência de seu papel no metabolismo. Porém, animais desenvolveram mecanismos para detectar e responder a esses produtos químicos. Melanomacrófagos (MMs) hepáticos são células que contém melanina e que estão envolvidos nesse processo, devido a sua função de detoxificação. Dessa forma, nossos objetivos foram avaliar os efeitos do BaP e/ou aNF nos MMs e efeitos genotóxicos dos compostos. Os animais foram expostos a 2mg/kg de BaP e/ou 20mg/kg de aNF por 48 horas e 7 dias. Após os experimentos o fígado passou por procedimentos histológicos para quantificação da área de melanina dos MMs e fagocitose na microscopia de luz; microscopia de fluorescência para análise do citoesqueleto dos MMs; espectrofotometria para quantificar a atividade EROD e síntese de melanina dos MMs; e análise genotóxica dos eritrócitos. Em peixes, após 7d houve diminuição da área de melanina, devido a uma inibição do BaP... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Fish and amphibians are vulnerable to various pollutants. Benzo[a]pyrene (BaP) is a contaminant with toxic properties. Metabolized by the enzyme P450 (CYP1A1) produces toxic byproducts. Known as a CYP1A1 inhibitor, α-naphtoflavone (aNF) prevents BaP metabolism and the generation of toxic metabolites. aNF is an exogenous compound and is not known for its effect on organs and tissues. The liver is the major source of catalyzing enzymes for biotransformation reactions. The susceptibility of the organ to damage by chemical agents is a consequence of its role in metabolism. However, animals have developed mechanisms to detect and respond to these chemicals. Melanomacrophages (MMs) are involved in this process due to their detoxification function. Thus, our objectives were to evaluate the effects of BaP and/or aNF on MMs and genotoxic effects of compounds. The animals were exposed to 2mg/kg BaP and/or 20mg/kg aNF for 48 hours and 7 days. After the experiments, the procedures for light microscopy, fluorescence, spectrophotometry and genotoxic analysis were followed. In fish, after 7d there was a decrease in melanin area, due to a inhibition of BaP in the melanogenic pathway; melanosome aggregation by interference in actin filaments; and increased frequency of micronucleus by the genotoxic potential of the compound. In anurans, at 48h, there was an increase in melanin production in BaP treatments, due to the antioxidant role of melanin; decreased phagocytosis by interfering with this... (Complete abstract click electronic access below) / Doutor