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TESE Kelma Sirleide De Souza.Pdf UNIVERSIDADE FEDERAL DE PERNAMBUCO CENTRO DE CIÊNCIAS BIOLÓGICAS PROGRAMA DE PÓS-GRADUAÇÃO EM CIÊNCIAS BIOLÓGICAS KELMA SIRLEIDE DE SOUZA Efeitos bioquímicos e genotóxicos de poluentes na ostra Crassostrea sp., na região norte do complexo estuarino Canal de Santa Cruz, litoral norte de Pernambuco Recife 2016 KELMA SIRLEIDE DE SOUZA Efeitos bioquímicos e genotóxicos de poluentes na ostra Crassostrea sp., na região norte do complexo estuarino Canal de Santa Cruz, litoral norte de Pernambuco Tese apresentada ao Programa de Pós- Graduação em Ciências Biológicas da Universidade Federal de Pernambuco como pré- requisito para a obtenção do grau de doutor em Ciências Biológicas. Orientador: Prof. Dr. Ranilson de Souza Bezerra Co- Orientador: Dr. Caio Rodrigo Dias de Assis Recife 2016 Catalogação na fonte Elaine Barroso CRB 1728 Souza, Kelma Sirleide de Efeitos bioquímicos e genotóxicos de poluentes na ostra Crassostrea sp. na região norte do complexo estuarino Canal de Santa Cruz, litoral norte de Pernambuco / Kelma Sirleide de Souza - Recife: O Autor, 2016. 91 folhas: il., fig., tab. Orientador: Ranilson de Souza Bezerra Coorientador: Caio Rodrigo Silva de Assis Tese (doutorado) – Universidade Federal de Pernambuco. Centro de Biociências. Ciências Biológicas, 2016. Inclui referências 1. Ostras 2. Estuários 3. Santa Cruz, Canal de (PE) I. Bezerra, Ranilson de Souza (orient.) II. Assis, Caio Rodrigo Silva de (coorient.) III. Título 594.4 CDD (22.ed.) UFPE/CB-2017- 406 KELMA SIRLEIDE DE SOUZA Efeitos bioquímicos e genotóxicos de poluentes na ostra Crassostrea sp., na região norte do complexo estuarino Canal de Santa Cruz, litoral norte de Pernambuco Tese apresentada ao Programa de Pós- Graduação em Ciências Biológicas da Universidade Federal de Pernambuco como pré- requisito para a obtenção do grau de doutor em Ciências Biológicas. Aprovado em 30/06/2016 BANCA EXAMINADORA Prof. Dr. Ranilson de Souza Bezerra / UFPE Prof. Dra. Juliett de Fátima Xavier / UFAL Prof. Dra. Mônica Lúcia Adam / UFPE Dra. Marina Marcuschi / UFPE Dra. Renata Cristina da Penha França / UFPE “A imaginação é mais importante que o conhecimento. O conhecimento é limitado. A imaginação envolve o mundo.” Albert Einstein Ao meu filho Álvaro Nascimento pelo amor que me impulsiona e direciona meu olhar para o futuro. AGRADECIMENTOS A Deus, pois tudo pude porque ele me fortaleceu. À minha família pelo amor incondicional, apoio e incentivo durante toda minha vida. Especialmente a meus pais Helena Costa e Severino Souza. Às minhas irmãs Katia Souza e Katiane Souza, por serem verdadeiros anjos na minha vida e na do meu filho. Muito obrigada pelo amor maternal com que cuidaram do “nosso bambino” durante as longas horas de pesquisa. À minha sogra Lourdes Carvalho pela amizade, cuidado intensivo com minha alimentação, dedicação e amor doado ao meu filho. À minha cunhada Kátia Nascimento pela amizade, dedicação e amor nos cuidados com o meu filho durante a finalização deste trabalho. Ao Professor Dr. Ranilson de Souza Bezerra pela oportunidade de ingressar em seu grupo de pesquisa científica, pela confiança, orientação e apoio durante o desenvolvimento deste trabalho. Ao meu Coorientador amigo (Caillou) Dr. Caio Dias pela coorientação, amizade, prestatividade e conhecimento compartilhado durante minha passagem pelo labenz e principalmente na realização deste projeto de pesquisa. Meu muito obrigada! À professora Dra. Mônica Adam, pelos ensinamentos de Genotoxicidade, parceria, simplicidade e por ter sido uma amiga durante todos momentos de orientação. À minha amiga Paula Rayane, companheira de altas horas de pesquisa, pela dedicação, disciplina, independência, parceria valiosa durante o desenvolvimento desse doutorado. Sem você minha amiga, tudo seria mais difícil. Obrigada por tudo flor! À minha amiga dona Luciene Trindade, por estar sempre disponível para realizar as coletas das ostras tão necessárias para o começo, desenvolvimento e fim desse doutorado. Espero retribuir cientificamente todo o esforço que você dedicou a mim e ao meu projeto de pesquisa. Ao meu amigo anjo Anderson Balbino por ser essa pessoa maravilhosa, prestativa, e divina que és, pelos ensinamentos de genotox e pelas longas horas contando células ao meu lado, o que fortaleceu ainda mais nossa amizade. Muito obrigada. Às minhas amigas Kaline Katiely e Marlyete Araújo, pela amizade, ajuda constante nos experimentos, pela simplicidade em aprendermos uma com as outras nesse caminho longo que se chama pesquisa. Amigas vocês foram anjos que surgiram durante o doutorado e que ficarão para sempre na minha vida. À minha eterna amiga Juliett Xavier pelo apoio incondicional durante minha vida acadêmica, pela acolhida em sua casa durante os experimentos com as ostras controle, pelo conhecimento compartilhado e momentos felizes e tristes vividos e superados juntas. À minha amiga Cibelly Marques pela amizade, alegria, gargalhada escandalosa e contagiante, e ajuda sempre que necessária. À minha amiga Werlinha Mendes, pela gargalhada maravilhosa, que faz uma falta danada no meu cotidiano, pela prestatividade, bondade e por me ajudar em muitos momentos na vida acadêmica. À Major Maria Laura pela compreenssão, amizade e apoio referente as atividades docentes. Ao Programa de Pós-Graduação em Ciências Biologicas pela oportunidade e transmissão de conhecimento. A todos que integram o LABENZ: Ana Cláudia, Andreia Cybelle, Amália Medeiros, Augusto Freitas, Caio Assis, Cleópatra Silva, Cyndy Mary, Daniela Campeche, Daniele Matias, Dárlio Teixeira, Diego Buarque, Fábio Marcel, Flávia Thuanne, Guilherme Firmino, Helane Costa, Ian Porto, Janilson Felix, Jéssica Vasconcelos, Juliana Interaminense, Juliana Santos, Juliett Xavier, Kaline Catiely, Karina Ribeiro, Karollina Lopes, Kelma Souza, Lidiane Cristina, Liliane Moreira, Luiz Swintiskas, Marina Marchuschi, Marlyete Chagas, Milena Márcia, Natália Albuquerque, Paula Rayane, Rafael David, Raquel Pereira, Renata Cristina, Renata Nascimento, Robson Coelho, Ruy Tenório, Thiago Cahú, Vagne Melo, Werlayne Mendes, pelo convívio e troca de experiências. Ao professor Rodrigo Torres do LAGEA, por tonar possível o desenvolvimento do capítulo referente à genotoxicidade. Aos funcionários do PPGCB e do departamento de bioquímica Adenilda Eugênia, Miron Oliveira, Albérico Espírito Santo, Sr. João Virgínio e D. Helena pela gentileza, atenção e prestatividade. Aos colegas e amigos da turma do Doutorado em Ciências Biológicas pela convivência, permuta de conhecimentos e pelos momentos de descontração. Aos amigos que com um simples gesto ou sorriso tornaram mais fácil de superar os desencontros da vida. Em especial a Marcela Filgueiras, Daniel Moreira, Ane Emanuelle, Gertrudes Melo, Lúcia Lira e Walquíria Nicácio pela amizade, apoio e torcida constante durante todos esses anos que nos conhecemos. Aos professores da banca avaliadora desta tese pelas sugestões e enriquecimento do trabalho (Dra. Marina Marcuschi, Dra. Mônica Adam, Dra. Juliett Xavier e Dra. Renata França). A todos aqueles que porventura não foram citados, mas que contribuíram para a realização deste trabalho. Ao CNPq e CAPES pelo financiamento do projeto. RESUMO O uso de biomarcadores enzimáticos e de genotoxicidade é uma importante ferramenta na avaliação da contaminação ambiental. Esta poluição, que em grande parte tem origem nas atividades humanas, é liberada no ambiente tendo como destino final, preferencialmente, os corpos d’água. No estuário Canal de Santa de Cruz a expansão das atividades industriais e turísticas , além do elevado crescimento populacional humano e sobrepesca, colocam em risco o delicado sistema ecológico deste local. Neste contexto, o uso do molusco bivalve Crassostrea sp. como bioindicador torna-se uma ferramenta útil e eficaz para diagnosticar os efeitos dos impactos antropogênicos nesse ambiente. Portanto, a presente tese avaliou a atividade de biomarcadores enzimáticos (AChE, BChE, CAT, SOD, amilase, pepsina, tripsina, quimotripsina) e não enzimáticos, GSSG, GSH, Malonaldeído em brânquias, vísceras e músculo adutor da ostra do mangue Crassostrea sp., e o ensaio de Genotoxicidade e mutagenicidade (ensaio cometa e MN) em hemócitos de ostras capturadas durante o período seco e chuvoso em três locais do Canal de Santa de Cruz (vila velha Itamaracá – PE); e em uma região referência (Depuradora/controle) em Penedo, AL. O impacto genotóxico foi confirmado pela detecção de microlesões no DNA por meio do ensaio cometa, e pela alta frequência de macrolesões (MN) nos hemócitos em relação as ostras controle. Diferenças significativas (p<0,05) entre ostras coletadas sazonalmente no estuário, e entre estas ostras e ostras controle, foram observadas nas frequências de MN nas coletas do período seco e chuvoso; no Índice de dano no período seco e na frequência de danos no período chuvoso para um ponto de coleta. No presente trabalho, a AChE presente em vísceras e brânquias de Crassostrea sp. foi caracterizada físico-química e cineticamente e foi exposta a pesticidas organofosforados, carbamatos e a íons. Todos os pesticidas usados mostraram efeito inibitório na atividade da AChE apresentando decréscimo significativo. Foi verificado a influência de alguns íons sobre a atividade da AChE nos dois tecidos estudados. A atividade da AChE e BChE de brânquias, vísceras e musculo adutor de ostra coletadas no estuário no período seco e chuvoso, foi significativamente (p<0,05) menor que as ostras depuradas. No presente estudo foram observadas diferenças sazonais significativas (p<0,05) e respostas sinérgicas para AChE e BChE, e nos
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