Detection Of An Untyped Strain Of Bovine Respiratory Syncytial Virus In A Dairy Herd

Bovine respiratory syncytial virus (BRSV) causes important lower respiratory tract illness in calves. According to F and G proteins genetic sequences, three BRSV subgroups have been reported and characterized in several countries, showing differences in its distribution. In Brazil, the virus is widely disseminated throughout the herds and the few characterized isolates revealed the solely occurrence of the subgroup B. This study describes the detection and characterization of an untyped BRSV strain from a twenty-days-old calf from a herd without clinical respiratory disease. Nasal swabs were analyzed by RT-nested PCR for the F and G proteins genes. One sample has amplified the F protein gene. Sequencing and subsequent phylogenetic reconstruction were accomplished, revealing that the strain could not be grouped with any other BRSV subgroups reported. This result may suggest that the BRSV is in constantly evolution, even in Brazil, where the vaccination is not a common practice. More detailed studies about BRSV characterization are necessary to know the virus subgroups distribution among the Brazilian herds to recommend appropriated immunoprophylaxis.35525392550Affonso, I.B., Gatti, S.P., Alexandrino, B., Oliveira, M.C., Medeiros, A.S.R., Buzinaro, M.G., Samara, S.I., Detection of antibodies against bovine respiratory syncytial virus (BRSV) in dairy cattle with different prevalences of bovine herpesvirus type 1 (BHV-1) in São Paulo State, Brazil (2011) Semina: Ciências Agrárias, 32 (1), pp. 295-300. , LondrinaAlmeida, R.S., Domingues, H.G., Spilki, F.R., Larsen, L.E., Hagglund, S., Belák, S., Arns, C.W., Circulation of bovine respiratory syncytial virus in Brazil (2006) Veterinary Record, 158 (18), pp. 632-634. , LondonAlmeida, R.S., Spilki, F.R., Roehe, P.M., Arns, C.W., Detection of Brazilian bovine respiratory syncytial virus strain by a reverse transcriptase-nestedpolymerase chain reaction in experimentally infected calves (2005) Veterinary Microbiology, 105 (2), pp. 131-135. , AmsterdamArns, C.W., Campalans, J., Costa, S.C.B., Domingues, H.G., D'Arce, R.C.F., Almeida, R.S., Characterization of bovine respiratory syncytial virus isolated in Brazil (2003) Brazilian Journal of Medical and Biological Research, 36 (2), pp. 213-218. , Ribeirão PretoBaker, J.C., Frey, M., Bovine respiratory syncytial virus (1985) Veterinary Clinics of North America: Food Animal Practice, 1 (2), pp. 259-272. , PhiladelphiaBidokhti, M.R.M., Travén, M., Ohlson, A., Zarnegar, B., Baule, C., Belák, S., Alenius, S., Liu, L., Phylogenetic analysis of bovine respiratory syncytial viruses from recent outbreaks in feedlot and dairy cattle herds (2012) Archives of Virology, 157 (4), pp. 601-607. , New YorkBunt, A.A., Milne, R.G., Sayaya, T., Verbeek, M., Vetten, H.J., Walsh, J.A., Paramyxoviridae (2005) Virus Taxonomy, Eigth Report of the International Committee on Taxonomy of Viruses, pp. 655-671. , In: FAUQUET, C. M.MAYO, M. A.MANILOFF, J.DESSELBERGER, U.BALL, L. A. (Ed.). London: Elsevier: Academic PressCampalans, J., Arns, C.W., Serological evidence of bovine respiratory syncytial virus in Brazil (1997) Virus Reviews and Research, 2 (1-2), pp. 50-56. , Belo HorizonteDomingues, H.G., Spilki, F.R., Arns, C.W., Detecção molecular e análise filogenética de vírus respiratório sincicial bovino (BRSV) em swabs e tecido pulmonar de bovinos adultos (2011) Pesquisa Veterinária Brasileira, 31 (11), pp. 961-966. , Rio de JaneiroFurze, J.M., Roberts, S.R., Wertz, G.W., Taylor, G., Antigenically distinct G glycoproteins of BRSV strains share a high degree of genetic homogeneity (1997) Virology, 231 (1), pp. 48-58. , New YorkFurze, J., Wertz, G., Lerch, R., Taylor, G., Antigenic heterogeneity of the attachment protein of bovine respiratory syncytial virus (1994) Journal of General Virology, 75 (2), pp. 363-370. , LondonGonçalves, I.P.D., Simanke, A.T., Jost, H.C., Hötzel, I., Dal Soglio, A., Moojen, V., Detection of bovine respiratory syncytial virus in calves of Rio Grande do Sul, Brazil (1993) Ciência Rural, 23 (3), pp. 389-390. , Santa MariaHall, T.A., BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT (1999) Nucleic Acids Symposium Series, 41, pp. 95-98. , OxfordLarsen, L.E., Bovine respiratory syncytial virus (BRSV): A review (2000) Acta Veterinaria Scandinavica, 41 (1), pp. 1-24. , CopenhagenLarsen, L.E., Tjornehoj, K., Viuff, B., Extensive sequence divergence among bovine respiratory syncytial viruses isolated during recurrent outbreaks in closed herds (2000) Journal of Clinical Microbiology, 38 (11), pp. 4222-4227. , WashingtonNettleton, P.F., Gilray, J.A., Caldow, G., Gidlow, J.R., Durkovic, B., Vilcek, S., Recent isolates of Bovine respiratory syncytial virus from Britain are more closely related to isolates from USA than to earlier British and current mainland European isolates (2003) Journal of Veterinary Medicine Series B-Infectious Diseases and Veterinary Public Health, 50 (4), pp. 196-199. , BerlinProzzi, D., Walravens, K., Langedijk, J.P., Daus, F., Kramps, J.A., Letesson, J.J., Antigenic and molecular analyses of the variability of bovine respiratory syncytial virus G glycoprotein (1997) Journal of General Virology, 78 (2), pp. 359-366. , LondonSchrijver, R.S., Daus, F., Kramps, J.A., Langedijk, J.P.M., Buijs, R., Middel, W.G.J., Taylor, G., Van Oirschot, J.T., Subgrouping of bovine respiratory syncytial virus strains detected in lung tissue (1996) Veterinary Microbiology, 53 (3-4), pp. 253-260. , AmsterdamSchrijver, R.S., Langedijk, J.P.M., Poel, V.D.M.W.H., Middel, W.G.J., Kramps, J.A., Van Oirschot, J.T., Antibody responses against the G and F proteins of bovine respiratory syncytial virus after experimental and natural infections (1996) Clinical and Diagnostic Laboratory Immunology, 3 (5), pp. 500-506. , WashingtonSchrijver, R.S., Langedijk, J.P.M., Middel, W.G.J., Kramps, J.A., Rijsewijk, F.A.M., Van Oirschot, J.T., A bovine respiratory syncytial virus strain with mutations in subgroup-specific antigenic domains of the G protein induces partial heterologous protection in cattle (1998) Veterinary Microbiology, 63 (2-4), pp. 159-175. , AmsterdamSpilki, F.R., Almeida, R.S., Domingues, H.G., D'Arce, R.C.F., Ferreira, H.L., Campalans, J., Costa, S.C.B., Arns, C.W., Phylogenetic relationships of Brazilian bovine respiratory syncyctial virus isolates and molecular homology modeling of attachment glycoprotein (2006) Virus Research, 116 (1-2), pp. 161-168. , AmsterdamSpilki, F.R., Arns, C.W., Vírus respiratório sincicial bovino (2008) Acta Scientiae Veterinariae, 36 (3), pp. 197-214. , Porto AlegreStine, L.C., Hoppe, D.K., Clayton, L.K., Sequence conservation in attachment glycoproteins and antigenic diversity among bovine respiratory syncytial virus isolates (1997) Veterinary Microbiology, 54 (3-4), pp. 201-221. , AmsterdamTamura, K., Dudley, J., Nei, M., Kumar, S., MEGA 4: Molecular evolutionary genetics analysis (MEGA) software version 4.0 (2007) Molecular Biology and Evolution, 24 (8), pp. 1596-1599. , ChicagoTaylor, G., Stott, E.J., Furze, J., Ford, J., Sopp, P., Protective epitopes on the fusion protein of respiratory syncytial virus recognized by murine and bovine monoclonal antibodies (1992) Journal of General Virology, 73 (9), pp. 2217-2223. , LondonTaylor, G., Thomas, L.H., Furze, J.M., Cook, R.S., Wyld, S.G., Lerch, R., Hardy, R., Wertz, G.W., Recombinant vaccinia viruses expressing the F, G or N, but not the M2, protein of bovine respiratory syncytial virus (BRSV) induce resistance to BRSV challenge in the calf and protect against the development of pneumonic lesions (1997) Journal of General Virology, 78 (12), pp. 3195-3206. , LondonThomas, L.H., Cook, R.S., Wyld, S.G., Furze, J.M., Taylor, G., Passive protection of gnotobiotic calves using monoclonal antibodies directed at different epitopes on the fusion protein of bovine respiratory syncytial virus (1998) Journal of Infectious Diseases, 177 (4), pp. 874-880. 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Vírus respiratório sincicial bovino: detecção por imunoistoquímica em tecidos de camundongos e bovinos usando AcM contra o vírus respiratório sincicial humano

An immunoistochemical (IHC) test was developed to detect bovine respiratory syncytial virus (BRSV) in cell cultures and tissues of experimentally infected mice and calves, using a commercial monoclonal antibody (Mab) against human respiratory syncytial virus (HRSV), as a less expensive alternative, instead of producing specific monoclonal antibodies to BRSV. Clinical samples from calves suffering respiratory disease were also submitted to this test. IHC detected BRSV antigens in mouse tracheas (3, 5 and 7 days post-infection) and lungs (5 and 7 days post-infection), and in one of three lungs from experimentally infected calves. Lungs samples from two naturally infected calves were tested and resulted positive for BRSV by the IHC test. These results suggest that this test may be used in the future for diagnosis as well as a useful tool to assess the distribution of BRSV infections in Brazilian herds.Desenvolveu-se um teste de imunohistoquímica (IHQ) para detecção do vírus respiratório sincicial bovino (BRSV) multiplicado em cultivo celular e em tecidos de camundongos e bezerros infectados experimentalmente, utilizando um anticorpo monoclonal comercial contra o vírus respiratório sincicial humano (HRSV), como uma alternativa para eliminar os custos de produção de anticorpos monoclonais específicos para o BRSV. Amostras clínicas de bezerros com sintomatologia respiratória foram analisadas. A técnica mostrou-se eficiente na detecção de antígenos do BRSV em traquéias (3, 5 e 7 dias pós-infecção) e pulmões (5 e 7 dias pós-infecção) dos camundongos infectados e em uma das três amostras de pulmões dos bezerros infectados experimentalmente. Amostras de pulmões de dois animais com infecção natural foram positivas para BRSV. Conclui-se que o teste de IHQ pode ser usado no diagnóstico das infecções por BRSV e na avaliação da distribuição dessas infecções nos rebanhos bovinos brasileiros.97398

Comparative Evaluation Of Conventional Rt-pcr And Real-time Rt-pcr (rrt-pcr) For Detection Of Avian Metapneumovirus Subtype A [comparação Entre As Técnicas De Rt-pcr Convencional E Rt-pcr Em Tempo Real Para A Detecção Do Metapneumovírus Aviários Subtipo A]

Avian metapneumovirus (AMPV) belongs to Metapneumovirus genus of Paramyxoviridae family. Virus isolation, serology, and detection of genomic RNA are used as diagnostic methods for AMPV. The aim of the present study was to compare the detection of six subgroup A AMPV isolates (AMPV/A) viral RNA by using different conventional and real time RT-PCR methods. Two new RT-PCR tests and two real time RT-PCR tests, both detecting fusion (F) gene and nucleocapsid (N) gene were compared with an established test for the attachment (G) gene. All the RT-PCR tested assays were able to detect the AMPV/A. The lower detection limits were observed using the N-, F- based RRT-PCR and F-based conventional RT-PCR (10 0.3 to 10 1 TCID 50 mL -1). The present study suggests that the conventional F-based RT-PCR presented similar detection limit when compared to N- and F-based RRT-PCR and they can be successfully used for AMPV/A detection.39514451451Arns, C.W., Hafez, M.H., (1992) Swollen Head Syndrome in Poultry Flocks in Brazil, pp. 81-84. , In: WESTERN POULTRY DISEASE CONFERENCE, 41., 1992, Sacramento, USA. Proceedings... Davis, CA: Conference & Event Services, University of CaliforniaBarik, S., Transcription of human respiratory syncytial virus genome RNA in vitro: Requirement of cellular factor(s) (1992) Journal of Virology, 66, pp. 6813-6818Bäyon-Auboyer, M.H., Comparison of F-, G- and N-based RT-PCR protocols with conventional virological procedures for the detection and typing of turkey rhinotracheitis virus (1999) Archives of Virology, 144 (6), pp. 1091-1109. , http://www.springerlink.com/content/vxcm8vulvfx3vmeh, Disponível em:, Doi: 10.1007/s007050050572Cecchinato, M., Design, validation, and absolute sensitivity of a novel test for the molecular detection of avian pneumovirus (2004) Journal of Veterinary Diagnostic Investigation, 16 (6), pp. 582-585Choi, J.H., Development of real-time PCR assays for detection and quantification of human bocavirus (2008) Journal of Clinical Microbiology, 42 (3), pp. 249-253. , http://www.journalofclinicalvirology.com/article/S1386-6532(08)00070-X, Disponível em:, Doi: 10.1016/j.jcv.2008.02.010Cook, J.K., Cavanagh, D., Detection and differentiation of avian pneumoviruses (metapneumoviruses) (2002) Avian Pathology, 31 (2), pp. 117-132D'arce, R.C., et al, Subtyping of new Brazilian avian metapneumovirus isolates from chickens and turkeys by reverse transcriptase-nested-polymerase chain reaction (2005) Avian Pathology, 34 (2), pp. 133-136Dani, M.A., Molecular characterization of Brazilian avian pneumovirus isolates: Comparison between immunochemiluminescent Southern blot and nested PCR (1999) Journal of Virological Methods, 79 (2), pp. 237-241. , http://www.sciencedirect.com/science?_ob=MImg&_imagekey=B6T96-3WJDTS 6-D-7&_cdi=5106&_user=687304&_orig=search&_coverDate=05% 2F31%2F1999&_sk=999209997&view=c&wchp=dGLbVlW-zSkzS&md5= 6b42016b9f823422152fecdd9a0d7060&ie=, Disponível em:, Doi: 10.1016/S0166-0934(99)00020-8Fauquet, C.M., (2005) Virus taxonomy: VIIIth Report of the International Committee on Taxonomy of Viruses, p. 1162. , Amsterdam: Elsevier AcademicFerreira, H.L., Inhibition of avian metapneumovirus (AMPV) replication by RNA interference targeting nucleoprotein gene (N) in cultured cells (2007) Antiviral Research, 74 (1), pp. 77-81. , http://www.sciencedirect.com/science?_ob=MImg&_imagekey=B6T2H-4MR1GV 1-1-1&_cdi=4919&_user=10&_orig=search&_coverDate=04%2F30 %2F2007&_sk=999259998&view=c&wchp=dGLzVtb-zSkWb&md5=a1d6 3aa0ca8ae7b4fa06307793e8edc5&ie=, Disponível em:, Doi: 10.1016/j.antiviral.2006.12.002Giraud, P., Turkey rhinotracheitis in France: Preliminary investigations on a ciliostatic virus (1986) Veterinary Record, 119 (24), pp. 606-607Gough, R.E., Avian pneumoviruses (2003) Diseases of Poultry, pp. 92-99. , In: SAIF, M. et al., Ames: Iowa StateGuionie, O., Laboratory evaluation of a quantitative real-time reverse transcription PCR assay for the detection and identification of the four subgroups of avian metapneumovirus (2007) Journal of Virological Methods, 139 (2), pp. 150-158. , http://www.sciencedirect.com/science?_ob=MImg&_imagekey=B6T96-4MFJJ1 2-1-3&_cdi=5106&_user=10&_orig=search&_coverDate=02%2F28 %2F2007&_sk=998609997&view=c&wchp=dGLzVtz-zSkWz&md5=70a2 de1148e26bf618e402c54144d25e&ie=, Disponível em:, Doi: 10.1016/j.jviromet.2006.09.022Juhasz, K., Easton, A.J., Extensive sequence variation in the attachment (G) protein gene of avian pneumovirus: Evidence for two distinct subgroups (1994) Journal of General Virology, 75 (PART 11), pp. 2873-2880. , http://vir.sgmjournals.org/cgi/reprint/75/11/2873, Disponível em:, Doi: 10.1099/0022-1317-75-11-2873Keightley, M.C., Real-time NASBA detection of SARS-associated coronavirus and comparison with real-time reverse transcription-PCR (2005) Journal of Medical Virology, 77 (4), pp. 602-608. , http://www3.interscience.wiley.com/journal/112137449, Disponível em:, Doi: 10.1002/jmv.20498Loisy, F., Real-time RT-PCR for norovirus screening in shellfish (2005) Journal of Virological Methods, 123 (1), pp. 1-7Maertzdorf, J., Real-time reverse transcriptase PCR assay for detection of human metapneumoviruses from all known genetic lineages (2004) Journal of Clinical Microbiology, 42, pp. 981-986. , http://jcm.asm.org/cgi/reprint/42/3/981, Disponível em:, Doi: 10.1128/JCM.42.3.981-986.2004Otsuki, K., Demonstration of serum-neutralising antibody to turkey rhinotracheitis virus in serum from chicken flocks in Japan (1996) Journal of Veterinary Medical Science, 58 (9), pp. 869-874Pabbaraju, K., Diagnosis and epidemiological studies of human metapneumovirus using real-time PCR (2007) Journal of Clinical Virology, 40 (3), pp. 186-192. , http://www.journalofclinicalvirology.com/article/PIIS1386653207002740, Doi:10.1016/j.jcv.2007.08.004Reed, J.I., Muench, H., A simple method for estimating fifth percent endpoints (1938) American Journal Hygiene, 27, pp. 493-49

A survey for maintenance of virulent newcastle disease virus-free area in poultry production in Brazil

In 2003, Brazil was recognized as a pathogenic Newcastle Disease Virus (NDV) strain-free country for commercial poultry. This research was conducted in Brazil between December 2003 and March 2005 to verify the maintenance of this virulent NDV-free status. Serum samples from 5,455 flocks for commercial poultry farms were collected, comprising 81,825 broiler chickens. The farms were located in nine states of the country, grouped in three geographic regions. Serological evidence of NDV infection was detected in 28.8% of the surveyed farms. However, all fifteen viruses isolated and identified as Newcastle Disease Virus (NDV) were characterized as nonpathogenic strains, based on the Intracerebral Pathogenicity Index. These results showed that Brazil preserves the virulent NDV-free status for commercial flocks.368375Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Characterization of bovine respiratory syncytial virus isolated in Brazil

This paper presents the first isolation of bovine respiratory syncytial virus in Brazil and its physicochemical, morphological and molecular characterization. The virus was isolated from 33 samples of nasotracheal secretions, successively inoculated into a Madin-Darby bovine kidney cell culture, which was characterized by physicochemical tests and morphological observation by electron microscopy. The Brazilian sample is an RNA pleomorphic, enveloped, thermolabile and non-hemagglutinating spicular virus. Reverse transcription, followed by nested polymerase chain reaction (nRT-PCR) assay was carried out using oligonucleotides B1, B2A, B3 and B4 for the fusion proteins (F) and B5A, B6A, B7A and B8 for the attachment protein (G). The nRT-PCR-F amplified a fragment of 481 bp corresponding to part of the gene that codes for protein F, whereas nRT-PCR-G amplified a fragment of 371 bp, in agreement with part of the G gene. The virus isolated from Brazilian samples in this study corresponded to the bovine respiratory syncytial virus, and RT-PCR proved to be useful for the diagnosis of bovine clinical samples.21321

Complete Genome Sequence Of A Vaccinal Newcastle Disease Virus Strain Isolated From An Owl (rhinoptynx Clamator)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)A Newcastle disease virus (NDV) was isolated in chicken embryonated eggs after detection by real-time reverse transcription- PCR (RRT-PCR) from a captive owl swab. The complete genome sequence of APMV-1/Rhinoptynx clamator/Brazil/22516/2009 (APMV-1, avian paramyxovirus type 1) was obtained using Illumina sequencing. Phylogenetic analysis of the complete genome classified the isolate within NDV class II genotype II. © 2016 Van Borm et al.46CNPq, Conselho Nacional de Desenvolvimento Científico e TecnológicoConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq