Uso de anticorpos policlonais, microscopia eletrônica e histologia para detecção de partículas semelhantes ao iridovírus em rãs-touro
DOI:
https://doi.org/10.20950/1678-2305.2016v42n3p523Palavras-chave:
Rana catesbeiana, FV3, imunoeletro microscopia, ranavírusResumo
Os iridovirus do gênero Ranavirus têm sido implicados no declínio dos anfíbios em todo o mundo afetando animais de vida livre e aqueles em cativeiro. O objetivo deste estudo foi detectar a presença de particulas semelhantes ao iridovírus em três ranários na região sudeste do Brasil, utilizando anticorpos policlonais primários, microscopia eletrônica de transmissão (MET) e achados histológicos. Os órgãos alvo foram o figado e os rins. Sessenta rãs-touro adultas (Lithobates catesbeianus) e sessenta girinos da mesma espécie foram usados para o estudo. A MET revelou a presença de partículas semelhantes ao iridovírus em tecido hepático utilizando a técnica de contrastação negativa. Os resultados positivos foram também observados por imunomicroscopia eletrônica e imunocitoquímica. As análises histológicas nas mesmas amostras evidenciaram hemorragia no fígado e corpúsculos de inclusão em hepatócitos, e degeneração glomerulotubular e necrose nos rins. Os métodos usados neste estudo foram altamente eficientes na detecção de partículas semelhantes ao iridovírus e possivel infecção por ranavirus.
Referências
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CHINCHAR V.G.; HYATT, A.; MIYAZAKI, T.; WILLIANS, T. 2009 Family Iridoviridae: Poor viral relations no longer. Current Topics in Microbiology and Immunology, 328: 123-170.
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DIAS, D.C.; De STÉFANI, M.V.; FERREIRA, C.M.; FRANí"¡A, F.M.; RANZANI-PAIVA, M.J.T.; SANTOS, A.A. 2010 Haematologic and immunologic parameters of bullfrogs, Lithobates catesbeianus, fed probiotics. Aquacluture Research, 41: 1064-1071.
DUFFUS, A.L.J.; PAULI, B.D.; WOZNEY, K.; BRUNETTI, C.R.; BERRIL, M. 2008 Frog virus 3- like infections in aquatic amphibian communities. Journal Wildlife Diseases, 44: 109-120.
FAO - Food and Agriculture Organization of the United Nations. Cultured Aquatic Species Information Programme 2014. U R L : < http://www.fao.org/fishery/culturedspecies/Rana_catesbeiana/en> accessed 2 setembro 2014.
FERREIRA, C.M.; PIMENTA, A.G.C.; PAIVANETO, J.S. 2002 Introdução í Ranicultura. Boletim Técnico do Instituto de Pesca , 33: 1-15.
FOX, S.F.; GREER, A.L.; TORRES-CERVANTES, R.; COLLINS, J.P. 2006 First case of ranavirusassociated morbidity and mortality in natural populations of the South American frog Atelognathus patagonicus. Diseases of Aquatic
organisms, 72: 87-92.
GALLI, L.; PEREIRA, A.; MARQUEZ, A.; MAZZONI, R.; 2006 Ranavirus detection by PCR in cultured tadpoles (Rana catesbeiana, Shaw 1802) from South America. Aquaculture, 257(1-4): 78-82.
GOLDSMITH, C.S. and MILLER, S.E. 2009 Modern uses of electron microscopy for detection of viruses. Clinical Microbiology Reviews, 22(4): 552-63.
GREEN, D.M. and CONVERSE, K.A. 2005 Diseases of amphibian eggs and embryos. In: Majumdar S.K., Huffman JE, Brenner F.J., Panah A.I. (eds) Wildlife diseases: landscape epidemiology, spatial distribution, and utilization of remote sensing technology. Pennsylvania Academy of Science, p. 118-130.
GREER, A.M.; BERRILL, M.; WILSON, P.J. 2005 Five amphibian mortality events associated with ranavirus infection in south central Ontario, Canada. Diseases of Aquatic Organisms, 67: 9-14.
HAYAT, M.A.; MILLER, S.E. 1990 Negative Staining. Mc. Graw-Hill Publ. Company, New York. Micron and Microscopica Acta, 22(3): 310.
HIPOLITO, M. and BACH, E.E. 2002 Patologias em rã touro (Rana catesbeiana Shaw 1802). Primeira revisão da bibliografia brasileira. Arquivos do Instituto Biológico de São Paulo, 69(2): 113-120.
HIPOLITO, M.; CATROXO, M.H.B.; MARTINS, A.M.C.R.P.F.; MELO, N.A.; PITUCO, E.M.; GALLETI, N.T.C.; RANZANI-PAIVA, M.J.T.; MOURIí"˜O, J.L.P.; FERREIRA, C.M. 2012 Detection of White Spot Syndrome Virus in Brazil using Negative Staining, Immunoelectron Microscopy and Immunocytochemistry Techniques. International Journal of Morphology, 30(2): 761-768.
HOVERMAN, J.T.; GRAY, M.J.; MILLER, D.L. 2010 Anuran susceptibilities to ranaviruses: role of species identity, exposure route, and a novel virus isolate. Diseases of Aquatic Organisms, 89: 97-107.
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KNUTTON, S. 1995 Electron microscopical methods in adhesion. Methods Enzymol, 253: 145-58. MAJJI, S.; LAPATRA, S.; LONG, S.M.; SAMPLE, R.; BRYAN, L.; SINNING, A.; CHINCHAR, G. 2006 Rana catesbeiana virus Z (RCV-Z): a novel
pathogenic ranavirus. Diseases of Aquatic Organisms, 73: 1-11.
MAZZONI, R.; MESQUITA, A.J.; FLEURY, L.F.F.; BRITO, W.M.E.D.; NUNES, I.A.; ROBERT, J.; MORALES, H., COELHO, A.S.G.; BARTHASSON, D.L.; GALLI, L.; CATROXO, M.H.B. 2009 Mass mortality associated with a frog virus 3- like Ranavirus infection in farmed tadpoles Rana catesbeiana from Brazil. Diseases Aquatic Organisms, 86: 181-191.
MILLER, D.L.; RAJEEV, S.; GRAY, M.J.; BALDWIN, C.A. 2007 Frog Vírus 3 Infection, Cultured American bullfrogs. Emerging Infectious Diseases, 13: 342í 343
OIE The World Organisation for Animal Health. 2011 Infection with ranavirus. URL <http://www.oie.int/fileadmin/Home/eng/Health_standards/aahc/2010/en_chapitre_1.8.2.pdf> Accessed jul 2014.
ROBERT, J.; GEORGE, E.; ANDINO, F.J.; CHEN, G. 2011 Waterborne infectivity of the Ranavirus frog virus 3 in Xenopus laevis. Virology, 417(2): 410-417
ROMANSIC, J.C.; HIGASHI, E.M.; DIEZ, K.A.; BLAUSTEIN, A.R. 2007 Susceptibility of Newlymetamorphosed Frogs to a Pathogenic Water Mould (Saprolegnia sp.). Herpetology Journal (17): 161-166.
ZUPANOVIC, Z.; MUSSO, C.; LOPEZ, G.; LOUREIRO, C.L.; HYATT, A.D.; HENGSTBERGER, S.; ROBINSON, A.J. 1998
Isolation and characterization of iridoviruses from the giant toad Bufo marinus in Venezuela. Diseases of Aquatic Organisms, 33: 1-9.
CHINCHAR V.G.; HYATT, A.; MIYAZAKI, T.; WILLIANS, T. 2009 Family Iridoviridae: Poor viral relations no longer. Current Topics in Microbiology and Immunology, 328: 123-170.
CONVERSE, K.A. and GREEN, D.M. 2005 Diseases of salamanders. In: Majumdar S.K., Huffman J.E, Brenner F.J., Panah A.I. (eds) Wildlife diseases: landscape epidemiology, spatial distribution, and utilization of remote sensing technology. Pennsylvania Academy of Science, p. 118-130.
DASZAK, P.; BERGER, L.; CUNNINGHAM, A.A.; HYATT, A.D.; GREEN, D.E.; SPEARE, R. 1999 Emerging infectious diseases and amphibian population declines. Emerging Infection Diseases, 5: 735-748.
DIAS, D.C.; De STÉFANI, M.V.; FERREIRA, C.M.; FRANí"¡A, F.M.; RANZANI-PAIVA, M.J.T.; SANTOS, A.A. 2010 Haematologic and immunologic parameters of bullfrogs, Lithobates catesbeianus, fed probiotics. Aquacluture Research, 41: 1064-1071.
DUFFUS, A.L.J.; PAULI, B.D.; WOZNEY, K.; BRUNETTI, C.R.; BERRIL, M. 2008 Frog virus 3- like infections in aquatic amphibian communities. Journal Wildlife Diseases, 44: 109-120.
FAO - Food and Agriculture Organization of the United Nations. Cultured Aquatic Species Information Programme 2014. U R L : < http://www.fao.org/fishery/culturedspecies/Rana_catesbeiana/en> accessed 2 setembro 2014.
FERREIRA, C.M.; PIMENTA, A.G.C.; PAIVANETO, J.S. 2002 Introdução í Ranicultura. Boletim Técnico do Instituto de Pesca , 33: 1-15.
FOX, S.F.; GREER, A.L.; TORRES-CERVANTES, R.; COLLINS, J.P. 2006 First case of ranavirusassociated morbidity and mortality in natural populations of the South American frog Atelognathus patagonicus. Diseases of Aquatic
organisms, 72: 87-92.
GALLI, L.; PEREIRA, A.; MARQUEZ, A.; MAZZONI, R.; 2006 Ranavirus detection by PCR in cultured tadpoles (Rana catesbeiana, Shaw 1802) from South America. Aquaculture, 257(1-4): 78-82.
GOLDSMITH, C.S. and MILLER, S.E. 2009 Modern uses of electron microscopy for detection of viruses. Clinical Microbiology Reviews, 22(4): 552-63.
GREEN, D.M. and CONVERSE, K.A. 2005 Diseases of amphibian eggs and embryos. In: Majumdar S.K., Huffman JE, Brenner F.J., Panah A.I. (eds) Wildlife diseases: landscape epidemiology, spatial distribution, and utilization of remote sensing technology. Pennsylvania Academy of Science, p. 118-130.
GREER, A.M.; BERRILL, M.; WILSON, P.J. 2005 Five amphibian mortality events associated with ranavirus infection in south central Ontario, Canada. Diseases of Aquatic Organisms, 67: 9-14.
HAYAT, M.A.; MILLER, S.E. 1990 Negative Staining. Mc. Graw-Hill Publ. Company, New York. Micron and Microscopica Acta, 22(3): 310.
HIPOLITO, M. and BACH, E.E. 2002 Patologias em rã touro (Rana catesbeiana Shaw 1802). Primeira revisão da bibliografia brasileira. Arquivos do Instituto Biológico de São Paulo, 69(2): 113-120.
HIPOLITO, M.; CATROXO, M.H.B.; MARTINS, A.M.C.R.P.F.; MELO, N.A.; PITUCO, E.M.; GALLETI, N.T.C.; RANZANI-PAIVA, M.J.T.; MOURIí"˜O, J.L.P.; FERREIRA, C.M. 2012 Detection of White Spot Syndrome Virus in Brazil using Negative Staining, Immunoelectron Microscopy and Immunocytochemistry Techniques. International Journal of Morphology, 30(2): 761-768.
HOVERMAN, J.T.; GRAY, M.J.; MILLER, D.L. 2010 Anuran susceptibilities to ranaviruses: role of species identity, exposure route, and a novel virus isolate. Diseases of Aquatic Organisms, 89: 97-107.
ICTV International Committee on Taxonomy of Viruses. 2011 Virus Taxonomy: 2011 Release
(current). URL: <http://ictvonline.org/virusTaxonomy.asp?version=2011> Accessed jul 2014.
KNUTTON, S. 1995 Electron microscopical methods in adhesion. Methods Enzymol, 253: 145-58. MAJJI, S.; LAPATRA, S.; LONG, S.M.; SAMPLE, R.; BRYAN, L.; SINNING, A.; CHINCHAR, G. 2006 Rana catesbeiana virus Z (RCV-Z): a novel
pathogenic ranavirus. Diseases of Aquatic Organisms, 73: 1-11.
MAZZONI, R.; MESQUITA, A.J.; FLEURY, L.F.F.; BRITO, W.M.E.D.; NUNES, I.A.; ROBERT, J.; MORALES, H., COELHO, A.S.G.; BARTHASSON, D.L.; GALLI, L.; CATROXO, M.H.B. 2009 Mass mortality associated with a frog virus 3- like Ranavirus infection in farmed tadpoles Rana catesbeiana from Brazil. Diseases Aquatic Organisms, 86: 181-191.
MILLER, D.L.; RAJEEV, S.; GRAY, M.J.; BALDWIN, C.A. 2007 Frog Vírus 3 Infection, Cultured American bullfrogs. Emerging Infectious Diseases, 13: 342í 343
OIE The World Organisation for Animal Health. 2011 Infection with ranavirus. URL <http://www.oie.int/fileadmin/Home/eng/Health_standards/aahc/2010/en_chapitre_1.8.2.pdf> Accessed jul 2014.
ROBERT, J.; GEORGE, E.; ANDINO, F.J.; CHEN, G. 2011 Waterborne infectivity of the Ranavirus frog virus 3 in Xenopus laevis. Virology, 417(2): 410-417
ROMANSIC, J.C.; HIGASHI, E.M.; DIEZ, K.A.; BLAUSTEIN, A.R. 2007 Susceptibility of Newlymetamorphosed Frogs to a Pathogenic Water Mould (Saprolegnia sp.). Herpetology Journal (17): 161-166.
ZUPANOVIC, Z.; MUSSO, C.; LOPEZ, G.; LOUREIRO, C.L.; HYATT, A.D.; HENGSTBERGER, S.; ROBINSON, A.J. 1998
Isolation and characterization of iridoviruses from the giant toad Bufo marinus in Venezuela. Diseases of Aquatic Organisms, 33: 1-9.
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2016-09-30
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