Glicina E Demais Componentes Da Formulação De Uso Comercial

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Glicina E Demais Componentes Da Formulação De Uso Comercial UNIVERSIDADE DE SÃO PAULO ESCOLA DE ENGENHARIA DE LORENA AMANDA GUIMARÃES MORAES CAMARGO AVALIAÇÃO DA TOXICIDADE CELULAR EM HUMANOS INDUZIDA POR N-(FOSFONOMETIL)GLICINA E DEMAIS COMPONENTES DA FORMULAÇÃO DE USO COMERCIAL Lorena 2018 AMANDA GUIMARÃES MORAES CAMARGO AVALIAÇÃO DA TOXICIDADE CELULAR EM HUMANOS INDUZIDA POR N-(FOSFONOMETIL)GLICINA E DEMAIS COMPONENTES DA FORMULAÇÃO DE USO COMERCIAL Trabalho de conclusão de curso apresentado à Escola de Engenharia de Lorena da Universidade de São Paulo como requisito final para conclusão da Graduação do curso de Engenharia Ambiental. Orientador: Prof. Dr Morun Bernardino Neto. Lorena 2018 AUTORIZO A REPRODUÇÃO E DIVULGAÇÃO TOTAL OU PARCIAL DESTE TRABALHO, POR QUALQUER MEIO CONVENCIONAL OU ELETRÔNICO, PARA FINS DE ESTUDO E PESQUISA, DESDE QUE CITADA A FONTE Ficha catalográfica elaborada pelo Sistema Automatizado da Escola de Engenharia de Lorena, com os dados fornecidos pelo(a) autor(a) Camargo, Amanda Guimarães Moraes A / Amanda Guimarães Moraes Camargo; orientador Morun Bernardino Neto. - Lorena, 2018. 55 p. Monografia apresentada como requisito parcial para a conclusão de Graduação do Curso de Engenharia Ambiental - Escola de Engenharia de Lorena da Universidade de São Paulo. 2018 1. Glifosato. 2. Toxicidade. 3. Herbicidas. 4. Saúde pública. I. Título. II. Neto, Morun Bernardino, orient. AGRADECIMENTOS À Deus, por estar sempre presente nos momentos abençoando e direcionando o melhor para minha vida. Aos meus pais, Nelson e Mônica, por me ensinarem a correr atrás dos meus sonhos e objetivos, por todo o suporte necessário para que eu pudesse realizar a faculdade e me dedicar integralmente aos estudos. Aos meus irmãos, Rodrigo e Rafaella, por serem referências pra mim. Ao meu namorado Guilherme, por toda paciência e apoio emocional. Ao meu orientador Prof Dr. Morun, pela paciência, pela dedicação, pela confiança no meu trabalho. Aos meus colegas do trabalho da UNESP, por me ajudarem a desenvolver a pesquisa, por todas as discussões científicas. À minha orientadora de Iniciação Científica, Gretta, e à minha amiga de estágio, Kely, pela confiança, pelos aprendizados. Agradecimento ao apoio dado pela Secretaria Municipal de Saúde da Prefeitura de Lorena, que disponibilizou as amostras de material biológico. RESUMO A toxicidade celular proveniente de pesticidas utilizados na agricultura brasileira é um indicador de grande importância para a saúde do indivíduo e à saúde pública. Por meio da avaliação da citotoxicidade de eritrócitos é possível inferir o potencial de causar danos e de quebra da homeostase que um determinado toxicante apresenta. Diversos pesticidas já proibidos em outros países por levarem sérios danos ao meio ambiente e aos seres humanos, continuam liberados no Brasil. Nesse contexto, destaca-se o glifosato (N-fosfonometil-glicina) o mais vendido no país e um dos campeões de vendas no mundo. Por apresentar DL50 considerado baixo, o glifosato é tido como pouco tóxico e seguro quando usado dentro da recomendação. No entanto os efeitos tóxicos produzidos por uma formulação comercial vão além do componente ativo sendo que estabilizantes, dispersantes e conservantes não são totalmente livres de ação tóxica. Este trabalho visou avaliar a toxicidade celular em eritrócitos humanos induzida pelo glifosato e demais componentes da formulação de uso comercial. Para tal, o comportamento da fragilidade osmótica de eritrócitos na presença e ausência do toxicante foi avaliado graficamente por meio da dependência de lise celular em função da osmolaridade do meio. Os gráficos obtidos foram ajustados a uma sigmoide de Boltzmann. Esse trabalho obteve uma correlação forte, inversa e significativa do aumento da concentração da Hemoglobina com a diminuição da estabilidade eritrocitária. Palavras-chave: herbicidas, toxicidade, glifosato, saúde pública. Sumário 1 INTRODUÇÃO .................................................................................... 9 2 REVISÃO DE LITERATURA ............................................................. 11 2.1 Regularização ................................................................................ 16 2.2 Toxicologia ..................................................................................... 18 2.2.1 Toxicologia em crianças ........................................................... 19 2.3 Formulação .................................................................................... 20 2.4 Glifosato ......................................................................................... 21 2.4.1 Indenização .............................................................................. 25 2.5 MECANISMO DE AÇÃO ................................................................ 25 2.5.1 Em Vegetais ............................................................................. 25 2.5.2 Em Animais .............................................................................. 25 2.6 FRAGILIDADE OSMÓTICA ERITROCITÁRIA (FOE) .................... 26 2.6.1 Estrutura da Membrana Plasmática ......................................... 26 2.6.2 Eritrócitos (células vermelhas) ................................................. 27 2.6.3 Teste de fragilidade osmótica eritrocitária ................................ 28 3 OBJETIVOS ...................................................................................... 30 3.1 Objetivo geral ................................................................................. 30 3.2 Objetivos específicos ..................................................................... 30 4 MATERIAL E MÉTODOS .................................................................. 31 4.1 Considerações éticas ..................................................................... 31 4.2 Amostras de material biológico ...................................................... 31 4.3 Reagentes ...................................................................................... 32 4.4 Determinação da estabilidade de eritrócitos humanos na presença de glifosato 32 4.5 Determinação das curvas de transição de hemólise ...................... 34 4.5.1 Determinação da concentração de glifosato ............................ 35 4.6 Determinação de variáveis bioquímicas e hematológicas .............. 36 4.7 Análise estatística dos dados experimentais .................................. 37 5 RESULTADOS .................................................................................. 38 5.1 Teste de fragilidade osmótica eritrocitária (FOE) ........................... 38 5.2 Análise descritiva exploratória ........................................................ 41 5.3 Teste de normalidade exploratória Shapiro Wilk ............................ 43 5.4 Teste de comparação de dois grupos de Wilcoxon ........................ 45 5.5 Matriz de correlações Teste de Spearman ..................................... 47 6 DISCUSSÃO ..................................................................................... 48 7 Considerações finais ......................................................................... 50 8 Agradecimento .................................................................................. 50 REFERÊNCIAS .......................................................................................... 51 LISTA DE FIGURAS Figura 1: Evolução da área plantada em milhões de hectares de arroz, feijão, cana de açúcar, soja e mandioca por ano. ........................................................... 13 Figura 2 : Os 10 ingredientes ativos mais vendidos. .................................. 22 Figura 3 - vendas de glifosato por ano ....................................................... 23 Figura 4: Partes típicas de uma molécula fosfolipídica representada em (a) esquematicamente e em (b) por meio de fórmula estrutural condensada. ........... 27 Figura 5: Gráfico de osmose e ilustração de osmose em eritrócitos .......... 29 Figura 6: Amostras de material biológico.................................................... 32 Figura 7:Bancada Experimental. ................................................................ 33 Figura 8: (a) Banho termostatizado a 37,5 ˚C; (b) Adição de 20 µL de sangue em todos os tubos. ................................................................................................ 33 Figura 9: Tubos eppendorfs com adição de sangue: (a) antes do banho termostatizado; (b) após o banho termostatizado. ................................................ 33 Figura 10: Centrifuga com capacidade máxima de 16 tubos: (a) Vista superior; (b) Equipamento aberto .......................................................................... 34 Figura 11: Leitura de sobrenadante no espectrofotômetro: (a) Leitura da absorbância em 540nm; (b) Cubeta e tubo de eppendorfs de uma amostra com lise; (c) Cubeta e tubo eppendorf de uma amostra sem lise. ....................................... 34 Figura 12: Glifosato e palavras chave. ....................................................... 50 7 LISTA DE GRÁFICOS Gráfico 1: Curva típica de dependência de lise eritrocitária em função da concentração de NaCl. .......................................................................................... 35 Gráfico 2: Curva de fragilidade eritrocitária induzida pela variação da concentração de Roundup e concentração de NaCl constante em 0,9 g/dL. ....... 36 Gráfico 3:Curva de hemólise das dez amostras analisadas: (a) sem a presença de glifosato; (b) com glifosato. ............................................................... 38 Gráfico 4:Curva de hemólise das três amostras
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