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Avaliação Da Toxicidade De Resíduos Industriais E Urbanos Aplicados Na Agricultura

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Avaliação Da Toxicidade De Resíduos Industriais E Urbanos Aplicados Na Agricultura UNIVERSIDADE ESTADUAL PAULISTA “JÚLIO DE MESQUITA FILHO” unesp INSTITUTO DE BIOCIÊNCIAS RIO CLARO PROGRAMA DE PÓS-GRADUAÇÃO EM CIÊNCIAS BIOLÓGICAS BIOLOGIA CELULAR E MOLECULAR AVALIAÇÃO DA TOXICIDADE DE RESÍDUOS INDUSTRIAIS E URBANOS APLICADOS NA AGRICULTURA CINTYA APARECIDA CHRISTOFOLETTI Tese apresentada ao Instituto de Biociências do Campus de Rio Claro, Universidade Estadual Paulista, como parte dos requisitos para a obtenção do título de Doutor em Ciências Biológicas (Biologia Celular e Molecular). Maio - 2013 UNIVERSIDADE ESTADUAL PAULISTA “JÚLIO DE MESQUITA FILHO” unesp INSTITUTO DE BIOCIÊNCIAS RIO CLARO PROGRAMA DE PÓS-GRADUAÇÃO EM CIÊNCIAS BIOLÓGICAS BIOLOGIA CELULAR E MOLECULAR AVALIAÇÃO DA TOXICIDADE DE RESÍDUOS INDUSTRIAIS E URBANOS APLICADOS NA AGRICULTURA CINTYA APARECIDA CHRISTOFOLETTI Orientadora: Profª Drª Carmem Silvia Fontanetti Christofoletti Tese apresentada ao Instituto de Biociências do Campus de Rio Claro, Universidade Estadual Paulista, como parte dos requisitos para a obtenção do título de Doutor em Ciências Biológicas (Biologia Celular e Molecular). Rio Claro Estado de São Paulo - Brasil Maio - 2013 iii iv v vi Para minha família, que compartilha todos os momentos da vida, a caminhada, as conquistas e realizações. A eles, por tudo que têm me ensinado. vii AGRADECIMENTOS Meus sinceros agradecimentos às instituições: À Universidade Estadual Paulista “Júlio de Mesquita Filho” (UNESP), Campus de Rio Claro. Ao Instituto de Biociências, Departamento de Biologia e aos Laboratórios de Citogenética, Histologia, Microscopia Eletrônica e Mutagênese Ambiental, por me acolher e fornecerem toda a estrutura necessária para o desenvolvimento deste trabalho. À Pró-Reitoria de Pós-Graduação e Pesquisa da UNESP. Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) pela bolsa de estudos concedida (processo: 2009/50578-3). À Companhia de Saneamento Básico do Estado de São Paulo (SABESP) - ETE Franca, ao Sr. Américo Sampaio, ao Sr. Rui César Rodrigues Bueno e ao Sr. Luciano Remale, pela autorização das coletas de lodo de esgoto e biossólido; à Usina Santa Lúcia (Araras/SP), por nos fornecer as amostras de vinhaça e ao meu pai, Almir José Christofoletti, pelas coletas destas. Ao engenheiro agrônomo Prof. Dr. Cássio Hamilton Abreu Junior, do CENA/USP pelo auxílio com os cálculos para a aplicação das amostras de lodo de esgoto, biossólido e vinhaça. Agradeço a Deus pela vida, pela oportunidade de desenvolver este trabalho, pela presença constante, pelo suporte em todos os momentos e por colocar em meu caminho pessoas com as quais pude contar ao longo desta jornada. Dentre elas, minha orientadora e amiga, Profª. Drª. Carmem S. Fontanetti Christofoletti – sem parentesco – exemplo de pessoa e profissional. Não tenho palavras para descrever o quanto sou grata pelos anos de orientação, paciência, convívio, conversas e risadas. Agradeço também a amizade, o carinho e a cumplicidade. Espero um dia ser 1/5 da professora e orientadora que és! “Podem esquecer o que você disse, mas eles nunca esquecerão como você as fez sentir” (Carol Buchner). Aos meus pais, Almir e Analice Christofoletti, pelo apoio e amor incondicional. Meus irmãos, Diego e Eduardo J. Christofoletti, minhas cunhadas, Francieli e Natalie, pelo apoio e incentivo. Aos meus queridos avós, Aristides Christofoletti, Olga e Isidoro Schnetes (in memorian) que sei que olham e torcem por mim de onde estão! Aos meus dois “cãopanheiros”, Ike e Lambari. viii Ao meu marido Juliano Liscia Pedroso de Figueiredo, por compartilhar e participar da realização de mais este trabalho. Obrigada pelo auxílio nas coletas e tranporte dos resíduos; por sempre me escutar, apoiar, incentivar e acreditar que eu era capaz. “O verdadeiro amor nunca se desgasta. Quanto mais se dá, mais se tem” (Antonie de Saint-Exupéry). Aos meus sogros, Itamar e José Figueiredo, minha cunhada Daniella, pelo incentivo e carinho. Aos professores do Programa de Pós-Graduação, área de Biologia Celular e Molecular, por terem contribuído para minha formação. Em especial à Profª. Drª. Maria Aparecida Marin-Morales, Profª. Drª. Patrícia Pasquali Parise Maltempi, Profª. Drª. Sanae Kasahara, Profª. Drª. Doralice Maria Cella (in memorian), Prof. Dr. Diogo Cavalcanti Cabral de Mello, Profª. Drª. Ana Maria Costa Leonardo, Profª. Drª. Maria Izabel Camargo, Profª Drª. Lúcia Regina Ribeiro, Prof. Dr. Odair C. Bueno e Prof. Dr. Osmar Malaspina. Aos funcionários do Departamento de Biologia, Lucila Segala Franco e Cristiane M. Mileo, pela amizade e carinho. Aos técnicos dos laboratórios Sandra Ap. Veloso, Gerson Mello Souza, Mônika Iamonte, Pablo H. Nunes e Antonio Yabuki, pelo auxílio indispensável. À técnica do CEA, Francisca de Assis Mattioli Gonçalves. Aos que no momento integram o grupo de pesquisa da Profª Drª Carmem, Ana Cláudia de Castro Marcato, Annelise Francisco, Cleiton Souza, Cristina Morerira de Souza, Dânia Elisa Christofoletti Mazzeo Morales, Janaína Pedro-Escher, Jorge Evangelista Correia, Julia Fernanda Urbano Marinho, Nilton Righetto Neto, Raphael Baston de Souza, Thays Guedes, Vinícius Daguano Gastaldi e Yadira Ansoar Rodríguez. “O que une uma equipe é quando um cobre as fraquezas do outro” (Phil Jackson). Obrigada por todo o auxílio no desenvolvimento deste trabalho, o convívio diário, as muitas reuniões com discussões produtivas, a companhia nos congressos, o carinho e a amizade! Vocês são todos muitos especiais e moram no meu coração!... e aos que já o fizeram e muito me ajudaram: Amanda Alfonso Batista, Danila Torres, Danielle Giuliano Perez, Frederico Rezes Biagini, Guilherme Thiago Maziviero, José Augusto de Oliveira David, Larissa Rosa Nogarol, Luciana Tendolini Brito, Tamaris Gimenez Pinheiro, Tatiana da Silva Souza, Tatiane Abe e Vanessa Vanderléia Merlini. Em especial, aos meus co-orientados: Annelise Francisco, Jorge Evagelista Correia e Amanda Alfonso Batista. Agradeço a oportunidade, a amizade e a confiança que depositaram em mim e em meu trabalho. Com vocês, eu mais pude aprender do que ensinar. “Feliz aquele que transfere o que sabe e aprende o que ensina” (Cora Coralina). ix Ao grupo de alunos da Profª Drª Maria Aparecida Marin Morales, Bruna de Campos Ventura Camargo, Cristiane Talhiaferro, Franco Dani, Jaqueline Bianchi Ambrósio, Jaqueline Ap. de Oliveira, Laís Sommagio, Lívia Loureiro, Márcia Miyuki Hoshina, Maria Tereza Pamplona, Matheus M. Roberto, Michele Bereta, Nádia Corroqué, Raquel Vaz Hara, Roberta Kleiner, Paula Suares Rocha, Thaís Cristina Casimiro Fernandes, Verônica Monteiro e Yaliana Tafurt. Agradeço o auxílio com o presente trabalho, a amizade e os momentos de descontração. A todos os amigos do laboratório de Citogenética e do Departamento de Biologia. Aos amigos e professores da graduação, do Centro Universitário Hermínio Ometto (UNIARARAS). Aos meus amigos que muito me escutaram e suportaram a minha preocupação com o desenvolvimento deste trabalho: família Trombini, Karina F. G. Joaquim e Jean W. Joaquim, Márcia C. Bortolin Arnosti, Leticia Vargas Bueno, Sandra Mattos e Daniela Schmidt. “O valor das coisas não está no tempo em que elas duram, mas na intensidade com que acontecem. Por isso, existem momentos inesquecíveis, coisas inexplicáveis e pessoas incomparáveis” (Fernando Pessoa). x “Dai-me Senhor, a perseverança das ondas do mar, que fazem de cada recuo, um ponto de partida para um novo avançar”. Cecília Meirelles “Estou sempre alegre, essa é a melhor maneira de vencer os obstáculos da vida”. Charles Chaplin xi RESUMO A degradação de solos provocada pelo mau gerenciamento das atividades humanas consiste em uma constante e crescente preocupação. O tratamento adequado e disposição final do grande volume de resíduos urbanos, industriais e agrícolas produzidos diariamente são um grande desafio à comunidade científica. Neste sentido, vem sendo pesquisadas inúmeras alternativas para o aproveitamento e destinação ambientalmente segura de tais resíduos, dentre as quais se destaca a aplicação destes como fertilizantes agrícolas. Tendo isso em vista, o presente trabalho teve por objetivo simular a dose de aplicação em cultura de cana-de- açúcar, de amostras de lodo de esgoto primário, biossólido e vinhaça de cana-de-açúcar, de acordo com a legislação brasileira vigente, e verificar seus potenciais tóxico, citotóxico, genotóxico e mutagênico, tanto das amostras brutas quanto de combinações destas com amostras de solo controle, antes e após o seu bioprocessamento por diplópodos. As amostras foram analisadas antes da exposição à diplópodos, designado por tempo 0 (t0 – momento da mistura) e após o bioprocessamento destas pelos animais (t30 – 30 dias após a exposição aos diplópodos). As análises químicas das amostras demonstraram diferentes concentrações de metais e macro/micronutrientes, evidenciando o bioprocessamento destes resíduos pelos animais. Para avaliar a toxicidade das diferentes combinações dos resíduos foram avaliadas a taxa de mortalidade e o comportamento dos indivíduos, e realizadas análises histológicas, histoquímicas e ultra-estruturais do intestino médio dos diplópodos, após 7, 30 e 90 dias de exposição. Os resultados obtidos nas avaliações histológica, histoquímica e ultra-estrutural, evidenciaram a toxicidade dos resíduos, observada por diferentes alterações como aumento nas taxas de renovação epitelial, espessamento do bordo em escova, acúmulo de grânulos citoplasmáticos nas células hepáticas dos animais e grupamentos de hemócitos por entre as células hepáticas, indicando um processo inflamatório severo, pois foram observadas ainda a presença de células
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