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. , ChicagoThompson, J.D., Higgins, D.G., Gibson, T.J., Clustal W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, positions-specific gag penalties and weight matrix choice (1994) Nucleic Acids Research, 22 (22), pp. 4673-4680. , OxfordValarcher, J.F., Schelcher, F., Bourhy, H., Evolution of bovine respiratory syncytial virus (2000) Journal of Virology, 74 (22), pp. 10714-10728. , WashingtonValarcher, J.F., Taylor, G., Bovine respiratory syncytial virus infection (2007) Veterinary Research, 38 (2), pp. 153-180. , Les UlisPoel, V.D.W., Brand, A., Kramps, J.A., Van Oirschot, J.T., Respiratory syncytial virus infections in human beings and in cattle (1994) Journal of Infectious Diseases, 29 (2), pp. 215-228. , ChicagoVilcek, S., Elvander, M., Ballagi-Pordány, A., Belák, S., Development of nested PCR assays for detection of bovine respiratory syncytial virus in clinical samples (1994) Journal of Clinical Microbiology, 32 (9), pp. 2225-2231. , 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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... 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Epidemiological And Genetic Characteristics Associated With The Severity Of Acute Viral Bronchiolitis By Respiratory Syncytial Virus

Objective: to assess the epidemiological and genetic factors associated with severity of acute viral bronchiolitis (AVB) by respiratory syncytial virus (RSV). Data source: the key words "bronchiolitis", "risk factor", "genetics" and "respiratory syncytial virus", and all combinations among them were used to perform a search in the PubMed, SciELO, and Lilacs databases, of articles published after the year 2000 that included individuals younger than 2 years of age. Data synthesis: a total of 1,259 articles were found, and their respective summaries were read. Of these, 81 were selected, which assessed risk factors for the severity of AVB, and were read in full; the 60 most relevant studies were included. The epidemiologic factors associated with AVB severity by RSV were prematurity, passive smoking, young age, lack of breastfeeding, chronic lung disease, congenital heart disease, male gender, ethnicity, viral coinfection, low weight at admission, maternal smoking during pregnancy, atopic dermatitis, mechanical ventilation in the neonatal period, maternal history of atopy and/or asthma during pregnancy, season of birth, low socioeconomic status, Down syndrome, environmental pollution, living at an altitude > 2,500 meters above sea level, and cesarean section birth. Conversely, some children with severe AVB did not present any of these risk factors. In this regard, recent studies have verified the influence of genetic factors on the severity of AVB by RSV. Polymorphisms of the TLRs, RANTES, JUN, IFNA5, NOS2, CX3CR1, ILs, and VDR genes have been shown to be associated with more severe evolution of AVB by RSV. Conclusion: the severity of AVB by RSV is a phenomenon that depends on the varying degrees of interaction among epidemiological, environmental, and genetic variables. © 2013 Sociedade Brasileira de Pediatria. Published by Elsevier Editora Ltda. 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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

Prevalence of newcastle disease virus in broiler chickens (Gallus gallus) in Brazil

This study was carried out during 2002/2003, aiming to determine the prevalence of virulent Newcastle disease virus strains (NDV) in Brazilian commercial poultry farms. Clinical samples were obtained from the Southeastern, Southern and Central-Western regions, which comprise the main area of the Brazilian poultry production. Serum samples and tracheal and cloacal swabs of 23,745 broiler chickens from 1,583 flocks, including both vaccinated chickens and those with no vaccination information, were tested for NDV using a diagnostic ELISA kit. The seropositivity was 39.1%, and the isolation percentage by flock varied from 1.0 to 7.6%, and by region from 6.5 to 58.4%. Higher isolation rates (74.3-83.3%) were obtained after three passages in embryonated chicken eggs. All isolates preliminarily identified as NDV were characterized as nonpathogenic strains, as their Intracerebral Pathogenicity Index (ICPI) was below 0.7. Based on results of this study, Brazil can claim a virulent NDV-free status for commercial flocks.349357Conselho 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