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Detecção De Vírus Entéricos Em Amostras De Água E Sua Relação Com a Saúde Humana

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Detecção De Vírus Entéricos Em Amostras De Água E Sua Relação Com a Saúde Humana Detecção de vírus entéricos em amostras de água e sua relação com a saúde humana Fernando Rosado Spilki Programa de Pós-Graduação em Qualidade Ambiental Laboratório de Microbiologia Molecular [email protected] Ci€ncias Ambientais • Multi/Inter/Transdisciplinaridade ––““AA teoriateoria ‚‚ cinzacinza,, masmas oo mundomundo llƒƒ forafora ‚‚ verdeverde”” ‚‚GoetheGoethe Interfaces da crise Ambiental POPULAÇÃO RECURSOS POLUIÇÃO NATURAIS Crescimento e estimativas Qualidade da água • Conjunto de características, componentes orgânicos e inorgânicos e a composição do estado da biota aquática (Chapman, 1992) • A qualidade da água depende de fatores naturais e da ação antrópica Qualidade da água • Fatores não-humanos: – Hidrogeoquímica das rochas – Processos atmosféricos • Evapotranspiração • Ventos – Deposição de matéria orgânica e nutrientes – Processos biológicos • Microbiota do solo e da água Qualidade da água • Efeitos da ação antrópica – Introdução direta ou indireta pelo homem de componentes bióticos ou abióticos, substâncias em geral e energia tem efeitos modificadores, normalmente deletérios sobre a qualidade da água Efeitos da ação antrópica sobre a qualidade da água As atividades humanas podem ter efeitos sobre os ecossistemas aquáticos: I. Biota aquática II. Saúde humana (gastroenterites, etc) III. Comprometimento de atividades econômicas e de subsistência (pesca) IV. Deterioração da qualidade da água e dos recursos hídricos para abastecimento, irrigação e indústria V. Perda do potencial de recreação, lazer e turismo Histórico dos problemas de qualidade da água Acesso à água não-poluída 1990-96 Manejo da água no ambiente urbano Manejo da água no ambiente rural Manejo da água Manejo ideal da água IIEGA – FEEVALE - ANINQAS Justificativa IIEGA – FEEVALE - ANINQAS Eutrofização acelerada ou natural (sucessão ecológica) Eutrofização Análise microbiológica a) Vírus b) Bactérias c) Protozoários e Helmintos Saneamento E.T.A. Portaria MS Nº 518 de 25/03/04 Estabelece os procedimentos e responsabilidades relativos ao controle e vigilância da qualidade da água para consumo humano e seu padrão de potabilidade , e dá outras providências. Tabela 8 - Número mínimo de amostras mensais para o controle da qualidade da água de sistema de abastecimento, para fins de análises microbiológicas, em função da população abastecida. PARÂMETRO SISTEMA DE DISTRIBUIÇÃO (REDE E RESERVATÓRIOS) População abastecida 5.000 a 20.000 a < 5.000 hab 20.000 hab 250.000 hab > 250.000 hab 105 + (1 para COLIFORMES 10 1 para cada 30 +( 1 para cada TOTAIS 500 hab. cada 2.000 5.000hab.) hab.) máximo de 1.000 NOTA: Na saída de cada unidade de tratamento devem ser coletadas, no mínimo, duas amostras semanais, recomendando-se a coleta de, pelo menos 4 amostras semanais. E os agentes virais “Devem ser monitorados?” COMO??? Gastroenterites virais Fernando Rosado Spilki Gastroenterites virais Transmissão fecal-oral; 2a. Causa de morbidade por doenças infecciosas no mundo (até 80% dos atendimentos nos países em desenvolvimento); 2 milhões de óbitos em crianças por ano no mundo (mesmo número de óbitos por HIV-AIDS) 450 milhões de reais /ano/Brasil, apenas em faltas parciais ao trabalho! Rotavírus Família Reoviridae Respiratory-Enteric-Ophan-virus (Sabin, 1959) Vírus não-envelopados Capsídeo protéico camada dupla Genomas com 10 a 12 segmentos de RNA de fita dupla Rotavírus Estáveis a extremos de temperatura a pH Alta resistência no ambiente Gênero Rotavirus Grupos Rotavirus A Rotavirus B Rotavirus C Rotavirus D Rotavirus E Rotavirus F Rotavirus G Rotavirus A - Histórico e características gerais • GARV; RV-A • Partículas detectadas em fezes de crianças com gastroenterite > anos 1970 • Diarréia em outras espécies animais suínos, bovinos, ovinos, equinos, caninos antropozoonose Classificação Arch Virol. 2008;153(8):1621-9. Epub 2008 Jul 5. Recommendations for the classification of group A rotaviruses using all 11 genomic RNA segments. Matthijnssens J, Ciarlet M, Rahman M, Attoui H, Bányai K, Estes MK, Gentsch JR, Iturriza-Gómara M, Kirkwood CD, Martella V, Mertens PP, Nakagomi O, Patton JT, Ruggeri FM, Saif LJ, Santos N, Steyer A, Taniguchi K, Desselberger U, Van Ranst M. Gx-P[x]-Ix-Rx-Cx-Mx-Ax-Nx-Tx-Ex-Hx VP7-VP4-VP6-VP1-VP2-VP3-NSP1-NSP2-NSP3-NSP4-NSP5/6 Gatroenterite por Rotavirus A em seres humanos Transmissão fecal-oral Crianças entre 6 meses de idade e idade pré-escolar (5 anos) Diarréia, vômito, desidratação Quadro leve ou inaparente em adultos Prevenção: Vacinação saneamento básico tem impacto limitado Parashar UD, Gibson CJ, Bresse JS, Glass RI. Rotavirus and severe childhood diarrhea. Emerg Infect Dis. 2006 Feb;12(2):304-6. Figure 2. Estimated global distribution of rotavirus-related deaths. Each dot represents 1,000 rotavirus-related deaths. > 80% EM PAÍSES EM DESENVOLVIMENTO Sazonalidade: diarreia de outono e iverno Diagnóstico laboratorial Quando conveniente: Aglutinação em látex Teste de antígenos virais (strip-tests) SDS-PAGE (corrida eletroforética de RNA em gel de poliacrilamida) RT-PCR > qRT-PCR Isolamento em cultivo celular pouco indicado, laborioso (tratamento com tripsina) e difícil Células MA104, HeLa Prevenção Vacinação por via oral (indução de IgA) Problemas iniciais Vacinas comercialmente disponíveis a | Rotarix is an attenuated human rotavirus vaccine made of a tissue-culture-adapted human P1A[8]G1, VP6 subgroup II and NSP4 geno-group B strain. b | RotaTeq is a bovine (WC3)–human reassortant vaccine composed of the five strains shown, each containing a human rotavirus gene encoding the VP7 neutralizing protein from different serotypes. Notably, in the WI79-9 and SC2-9 viruses (the last was used to create the first), genes 3 (VP3) and 9 (VP7) are of human origin. Although VP6 and NSP4 can potentially be the targets of protective antibodies (Fig. 2), their role in immunity against disease in humans is unknown. Efeitos sobre epidemiologia Efeitos da vacinação HM123819(12220_06RJ) HM066140(14919_08RS) HM123821(12343_06RJ) HM123825(12589_06RJ) HM123826(12647_06RJ) HM123839(15894_08ES) 57 HM066133(12549_06RJ) HM066136(13158_06ES) Figure. Evolutionary HM066137(13668_07AL) relationships of VP6 gene from HM066138(13788_07SE) Human-GARV The evolutionary history was 50 HM066141(15311_08RJ) inferred using the Neighbor- HM066144(15786_08ES) Joining method [1]. The optimal HM066157(16064_09MA) tree with the sum of branch length = 0.22115913 is shown. HM066156(15863_08MA) The percentage of replicate trees CRIANCA-5 Genotype I2 in which the associated taxa CRIANCA-3 clustered together in the bootstrap test (500 replicates) 61 HM467951(LB2772) are shown next to the branches HM467947(LB2744) [2]. The tree is drawn to scale, HM066142(15771_08PE) with branch lengths in the same units as those of the HM066132(12482_06BA) evolutionary distances used to HM066131(12287_06BA) infer the phylogenetic tree. The HM066130(11837_06AC) evolutionary distances were computed using the Kimura 2- HM066128(11580_05AC) parameter method [3] and are in HM066127(11860_06RJ) the units of the number of base substitutions per site. The HM066126(11257_05MS) analysis involved 34 nucleotide 65 HM066135(13039_06RS) sequences. All positions HM123836(14397_07MA) containing gaps and missing data were eliminated. There were a HM066151(15958_08RS) total of 165 positions in the final 65 HM066146(15840_08RS) dataset. Evolutionary analyses GU199507(Matlab36-02) were conducted in MEGA5 [4]. EU372748(CMH150/01) 99 EU372749(CMH185/01) Genotype I1 + non (I+II) 69 GU199521(Dhaka6) EU372750(CMH186/01) 0.02 Norovírus • Fam€lia Caliciviridae – RNA fita simples, polaridade +, n•o-envelopado – Norovirus • gastroenterite • Principal causa de diarreia viral em seres humanos • Sintomas e epidemiologia semelhantes aos descritos hoje desde 1929 na literatura • Norwalk/Ohio/USA > Kapikian, 1958 • ‘‘small round structured viruses’’ (SRSV) – Sapovirus • gastroenterite • Sapporo, Jap•o, 1977 (Chiba, 1978) Morfologia Distribuição sazonal Norovírus Doen„a em adultos “diarreia dos cruzeiros”, surtos em escolas e locais de trabalho Gastroenterite aguda normalmente moderada (raramente grave) Perda de fluidos Les‡es do epitˆlio intestinal Explicação para a correlação entre os norovírus e os navios de cruzeiro (CDC) As doenças ocorridas em navios de cruzeiro contam com acompanhamento por organizações oficiais de saúde. Portanto, surtos são descobertos e relatados mais rapidamente num navio de cruzeiro do que em terra firme. Camarotes e instalações apertadas podem aumentar o contato entre grupos humanos. Passageiros recém-chegados podem trazer o vírus para os outros passageiros e para a tripulação. Como os norovírus se espalham (CDC) As pessoas podem ser infectadas com o vírus: Ao comer alimentos ou beber líquidos infectados com os norovírus Ao tocar superfícies ou objetos infectados com os norovírus e depois tocar a própria boca, nariz ou olhos Ao ter contato de pessoa a pessoa (com uma pessoa infectada) ao: estar presente enquanto alguém está vomitando dividir comida ou comer usando os mesmos talheres cuidar de uma pessoa doente apertar as mãos de alguém Deixando de lavar as mãos depois de usar o banheiro ou trocar fraldas, e antes de comer ou preparar alimentos. Adenovírus Família Adenoviridae DNA fita dupla 50 sorotipos em seres humanos AdV F - G> entéricos AdV-40 e -41 Excreção também de AdV respiratório AdV-2 e -5 Robert Green was born in 1895 in Wadena, Minnesota. He earned four degrees from the University of Minnesota, including an MD in 1922. He joined the staff
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