Use of polyclonal antibodies, electron microscopy and histopathology to detect iridovirus-like particles in bullfrogs
DOI:
https://doi.org/10.20950/1678-2305.2016v42n3p523Keywords:
Rana catesbeiana, FV3, immunoelectron microscopy, ranavirusAbstract
Iridoviruses of the Ranavirus genus have been implicated in the decline in amphibians worldwide, capable of affecting animals b o t h i n t h e w i l d a n d i n c a p t i v i t y . This study a i m e d to detect iridovirus-like particles from three frog farms in southeastern Brazil using primary polyclonal antibodies, transmission electron microscopy (TEM) and histologic findings. The target organs were liver and kidneys. Sixty adults and sixty tadpoles of bullfrogs (Lithobates catesbeianus) were used in the study. TEM revealed the presence of iridovirus-like particles in hepatic tissue using the negative staining technique. Positive results were also observed by immunoelectron microscopy and immunocytochemistry (ICC). The histological analysis o f t h e s a m p l e s s h o w e d liver hemorrhage and corpuscles inclusion in hepatocytes as well as glomerulotubular degeneration and necrosis in the kidneys. The methods used in this study were highly efficient to detect the presence of iridovirus-like particles and possible infection of ranavirus.
References
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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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