PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Medeiros T.N.S., Lorenzetti E., Alfieri A.F. & Alfieri A.A. 2014. Severe diarrhea outbreak in beef calves (Bos indicus) caused by G6P[11], an emergent genotype of bovine rotavirus group A. Pesquisa Veterinária Brasileira 34(8):717-722. Laboratório de Virologia Animal, Departamento de Medicina Veterinária Preventiva, Universidade Estadual de Londrina, Rodovia Celso Garcia Cid, Campus Universitário, Cx. Postal 10011, Londrina, PR 86057-970, Brazil. E-mail: alfieri@uel.br The episodes of diarrhea caused by neonatal bovine rotavirus group A (BoRVA) constitute one of the major health problems in the calf rearing worldwide. The main G (VP7) and P (VP4) genotypes of BoRVA strains involved in the etiology of diarrhea in calves are G6P[1], G10P[11], G6P[5], and G8P[1]. However, less frequently, other G and P genotypes have been described in BoRVA strains identified in diarrheic fecal samples of calves. This study describes the identification and molecular characterization of an emerging genotype (G6P[11]) in BoRVA strains involved in the etiology of a diarrhea outbreak in beef calves in a cattle herd of high production in extensive management system. The diarrhea outbreak, which showed high morbidity (60%) and lethality (7%) rates, occurred in calves (n= 384) Nelore (Bos indicus) up to 30-day-old from the State of Mato Grosso do Sul, Brazil. BoRVA was identified in 80% (16/20) of the fecal samples analyzed by polyacrylamide gel electrophoresis (PAGE) technique. In all PAGE-positive fecal samples were amplified products with 1,062-bp and 876-bp in the RT-PCR assays for VP7 (G type) and VP4 (VP8*) (P type) of BoRVA, respectively. The nucleotide sequence analysis of VP7 and VP4 genes of four wild-type BoRVA strains showed G6-III P[11]-III genotype/lineage. The G6P[11] genotype has been described in RVA strains of human and animal hosts, however, in calves this genotype was only identified in some cross-sectional studies and not as a single cause of diarrhea outbreaks in calves with high morbidity and lethality rates as described in this study. The monitoring of the G and P genotypes of BoRVA strains involved in diarrhea outbreaks in calves is important for both animal and public health by allowing the identification of the most frequent genotypes, the characterization of novel genotypes and to identify reassortments with genotypes described in animal and human hosts. The results of this study show the importance of the monitoring of the genotypes of BoRVA strains involved in episodes of bovine neonatal diarrhea as for characterization of frequency of occurrence and pathogenic potential of uncommon genotypes as for monitoring of the emergency of different BoRVA genotypes not included in commercial vaccines.

• Calf rotaviruses bezerro rotavirose

Abstract in Portuguese:

RERSUMO.- Medeiros T.N.S., Lorenzetti E., Alfieri A.F. & Alfieri A.A. 2014. Severe diarrhea outbreak in beef calves (Bos indicus) caused by G6P[11], an emergent genotype of bovine rotavirus group A. [Surto de diarreia neonatal em bezerros de corte (Bos indicus) ocasionado por um genotipo emergente G6P[11] de rotavírus bovino grupo A.] Pesquisa Veterinária Brasileira 34(8):717-722. Laboratório de Virologia Animal, Departamento de Medicina Veterinária Preventiva, Universidade Estadual de Londrina, Rodovia Celso Garcia Cid, Campus Universitário, Cx. Postal 10011, Londrina, PR 86057-970, Brazil. E-mail: alfieri@uel.br Os episódios de diarreia neonatal ocasionados pelo rotavírus bovino grupo A (BoRVA) constituem-se em um dos principais problemas sanitários na criação de bezerros em todo o mundo. Os principais genotipos G (VP7) e P (VP4) de cepas de BoRVA envolvidos na etiologia da diarreia em bezerros são G6P[1], G10P[11], G6P[5] e G8P[1]. No entanto, com menor frequência, outros genotipos G e P têm sido descritos em cepas de BoRVA identificadas em amostras de fezes diarreicas de bezerros. Este estudo descreve a identificação e caracterização molecular de um genotipo emergente (G6P[11]) em cepas de BoRVA envolvidas na etiologia de um surto de diarreia em bezerros de um rebanho bovino de corte de alta produção em sistema de manejo extensivo. O surto, que apresentou altas taxas de morbidade (60%) e de letalidade (7%), ocorreu em bezerros (n=384) da raça Nelore (Bos indicus) com até 30 dias de idade, provenientes do estado do Mato Grosso do Sul, Brasil. O BoRVA foi identificado em 80% (16/20) das amostras fecais analisadas pela técnica de eletroforese em gel de poliacrilamida (PAGE). Em todas as amostras fecais PAGE-positivas foi possível a amplificação por RT-PCR de produtos com 1.062 pb e 876 pb referentes aos genes VP7 (G tipo) e VP4 (VP8*) (P tipo), respectivamente, de BoRVA. A análise da sequência de nucleotídeos dos genes VP7 e VP4 de quatro cepas de BoRVA demonstrou a presença do genotipo/linhagem G6-III P[11]-III. O genotipo G6P[11] tem sido descrito em cepas de RVA de hospedeiros humanos e animais. Contudo, em bezerros, este genotipo foi apenas identificado em alguns estudos transversais e não como a única causa de surtos de diarreia em bezerros com altas taxas de morbidade e letalidade como descrito neste estudo. O monitoramento dos genotipos G e P de cepas de BoRVA envolvidos em surtos de diarreia em bezerros é relevante tanto para a saúde animal quanto para a saúde pública por possibilitar a identificação dos genotipos mais frequentes, a caracterização de novos genotipos e por identificar reassortments com genotipos descritos em hospedeiros humanos e animais. Os resultados deste estudo demonstram a importância do monitoramento dos genotipos de cepas de BoRVA envolvidas em surtos de diarreia neonatal bovina tanto para a caracterização da frequência de ocorrência e potencial patogênico de genotipos incomuns quanto para o monitoriamento da emergência de genotipos distintos daqueles incluídos em vacinas comerciais.

Abstract in English:

ABSTRACT.- Lorenzetti E., Stipp D.T., Possatti F., Campanha J.E.T., Alfieri A.F. & Alfieri A.A. 2014. Diarrhea outbreaks in suckling piglets due to rotavirus group C single and mixed (rotavirus groups A and B) infections. Pesquisa Veterinária Brasileira 34(5):391-397. Laboratory of Animal Virology, Department of Veterinary Preventive Medicine, Universidade Estadual de Londrina, Rodovia Celso Garcia Cid, Campus Universitário, Cx. Postal 10011, Londrina, PR 86057-970, Brazil. E-mail: alfieri@uel.br Porcine group A rotavirus (PoRVA) is a major cause of neonatal diarrhea in suckling and recently weaned piglets worldwide. The involvement of non-group A rotavirus in cases of neonatal diarrhea in piglets are sporadic. In Brazil there are no reports of the porcine rotavirus group C (PoRVC) as etiologic agent of the diarrhea outbreaks in piglets. The aim of this study was to describe the identification of rotavirus group C in single and in mixed infection with rotavirus groups A and B in three neonatal diarrhea outbreaks in suckling (≤21-day-old) piglets, with 70% to 80% and 20% to 25% of morbidity and lethality rates, respectively, in three pig herds located in the state of Santa Catarina, Brazil. The diagnosis of PoRV in the diarrheic fecal samples was performed using polyacrylamide gel electrophoresis (PAGE) to identify the presence of porcine rotavirus groups A, B (PoRVB), and C, and by RT-PCR (PoRVA and PoRVC) and semi-nested (SN)-PCR (PoRVB) to partially amplify the VP4 (VP8*)-VP7, NSP2, and VP6 genes of PoRVA, PoRVB, and PoRVC, respectively. One RT-PCR (PoRVA and PoRVC) and SN-PCR (PoRVB) product of each group of rotavirus of each diarrhea outbreak was submitted to nucleotide (nt) sequence analysis. Based on the PAGE technique, 4 (25%) and 1 (6.25%) of the 16 diarrheic fecal samples evaluated in the first outbreak presented PoRVA and PoRVC electropherotype, respectively, and 11 (68.75%) were negative. In the second outbreak, 3 (42.85%) of the 7 fecal samples evaluated presented PoRVA electropherotype, and in 3 (42.85%) and in 1 (14.3%) fecal samples were detected inconclusive and negative results, respectively. Three (30%) of the 10 fecal samples of the third outbreak presented PoRVC electropherotype; 5 (50%) and 2 (20%) samples showed negative and inconclusive results, respectively. Based on the RT-PCR and SN-PCR assays in the first neonatal diarrhea outbreak, PoRVC was detected in 13 (81.2%) of the 16 diarrheic fecal samples evaluated. PoRVC single infection was identified in 4 (25%) of these samples and mixed infections with PoRVA and PoRVB in 9 (56.2%) fecal samples. All of the seven diarrheic fecal samples evaluated from the second neonatal diarrhea outbreak were positive for PoRVC, whereas its mixed infection with other PoRV groups was detected in 4 (57.2%) samples. In the third outbreak, PoRVC in single infection was detected in all of the 10 diarrheic fecal samples analyzed. In the nt sequence analysis, the PoRVA strains of the first and second outbreaks demonstrated higher nt identity with G4P[6] and G9P[23] genotypes, respectively. The PoRVB strains (first and second outbreaks) and the PoRVC strains (first, second, and third outbreaks) showed higher nt identity and clustered in the phylogenetic tree with PoRVB and PoRVC strains that belong to the N4 and I1 genotypes, respectively. This is the first description in Brazil of the involvement of PoRVC in the etiology of diarrhea outbreaks in suckling piglets. The results of this study demonstrated that PoRVC, in both single and mixed infections, is an important enteropathogen involved in neonatal diarrhea outbreaks in piglets and that the use of more sensitive diagnostic techniques allows the identification of mixed infections involving two or even three groups of PoRV, which may be more common than previously reported.

• Diarrhea rotaviruses diarreia rotavirose

Abstract in Portuguese:

RESUMO.- Lorenzetti E., Stipp D.T., Possatti F., Campanha J.E.T., Alfieri A.F. & Alfieri A.A. 2014. Diarrhea outbreaks in suckling piglets due to rotavirus group C single and mixed (rotavirus groups A and B) infections. [Surtos de diarreia em leitões lactentes por rotavírus grupo C em infecções singulares e mistas (rotavirus grupos A e B).] Pesquisa Veterinária Brasileira 34(5):391-397. Laboratory of Animal Virology, Department of Veterinary Preventive Medicine, Universidade Estadual de Londrina, Rodovia Celso Garcia Cid, Campus Universitário, Cx. Postal 10011, Londrina, PR 86057-970, Brazil. E-mail: alfieri@uel.br O rotavírus suíno grupo A (PoRVA) é uma das principais causas de diarreia neonatal em leitões lactentes e recém-desmamados em todo o mundo. As descrições do envolvimento de rotavírus não-grupo A em quadros de diarreia neonatal em leitões são esporádicas. No Brasil não há relatos do envolvimento do rotavírus suíno grupo C (PoRVC) na etiologia dos surtos de diarreia em leitões. O objetivo deste estudo foi descrever a identificação de rotavírus grupo C em infecções singulares e mistas com os rotavírus grupos A e B em três surtos de diarreia neonatal em leitões lactentes (≤21 dias de idade), com taxas de morbidade de 70% a 80% e de letalidade de 20% a 25%, em três rebanhos suínos localizados no estado de Santa Catarina, Brasil. O diagnóstico de PoRV nas amostras de fezes diarreicas foi realizado por eletroforese em gel de poliacrilamida (PAGE) para identificar a presença dos grupos A, B (PoRVB), e C de rotavírus suíno e por RT-PCR (PoRVA e PoRVC) e semi-nested (SN)-PCR (PoRVB) com a amplificação parcial dos genes VP4 (VP8*)-VP7, NSP2 e VP6 de PoRVA, PoRVB e PoRVC, respectivamente. Um produto de RT-PCR (PoRVA e PoRVC) e SN-PCR (PoRVB) de cada grupo de rotavírus de cada um dos três surtos de diarreia foi submetido à análise da sequência de nucleotídeos (nt). Com base na técnica de PAGE, 4 (25%) e 1 (6,25%) das 16 amostras de fezes analisadas no primeiro surto apresentaram eletroferotipo característico de PoRVA e PoRVC, respectivamente, e 11 (68,75%) amostras fecais foram negativas. No segundo surto, 3 (42,85%) das 7 amostras de fezes analisadas apresentaram perfil eletroforético de PoRVA; em 3 (42,85%) e em 1 (14,3%) amostras de fezes foram detectados resultados inconclusivos e negativos, respectivamente. Três (30%) das 10 amostras de fezes do terceiro surto apresentaram eletroferotipo característico de PoRVC; 5 (50%) e 2 (20%) amostras apresentaram resultados negativos e inconclusivos, respectivamente. Com base nos resultados da RT-PCR e SN-PCR, no primeiro surto de diarreia neonatal o PoRVC foi detectado em 13 (81,2%) das 16 amostras de fezes diarreicas analisadas, sendo que em 4 (25%) amostras foi identificada infecção singular e em 9 (56,2%) amostras infecção mista com PoRVA e PoRVB. Todas as sete amostras de fezes diarreicas provenientes do segundo surto de diarreia neonatal foram positivas para o PoRVC, enquanto infecções mistas com outros grupos de PoRV foram detectadas em 4 (57,2%) amostras. No terceiro surto o PoRVC foi detectado em infecção singular em todas as dez amostras de fezes diarreicas analisadas. Na análise da sequência de nt as cepas de PoRVA do primeiro e segundo surtos demonstraram maior identidade de nt com os genotipos G4P[6] e G9P[23], respectivamente. As cepas de PoRVB (primeiro e segundo surtos) e as cepas de PoRVC (primeiro, segundo e terceiro surtos) mostraram maior identidade de nt com cepas de PoRVB e PoRVC que pertencem aos genotipos N4 e I1, respectivamente. Esta é a primeira descrição realizada no Brasil do envolvimento de PoRVC na etiologia de surtos de diarreia em leitões lactentes. Os resultados deste estudo demonstram que o PoRVC, tanto em infecções singulares quanto em infecções mistas, é um importante enteropatógeno envolvido em surtos de diarreia neonatal em leitões e que o uso de técnicas de diagnóstico mais sensíveis permite caracterizar que infecções mistas, com dois ou até mesmo com três grupos de PoRV, podem ser mais comuns do que anteriormente relatado.

Abstract in English:

ABSTRACT.- Leme R.A., Alfieri A.F. & Alfieri A.A. 2013. Torque teno sus virus (TTSuV) infection at different stages of pig production cycle. Pesquisa Veterinária Brasileira 33(7):840-846. Laboratório de Virologia Animal, Departamento de Medicina Veterinária Preventiva, Universidade Estadual de Londrina, Rodovia Celso Garcia Cid, Campus Universitário, Cx. Postal 10011, Londrina, PR 86057-970, Brazil E-mail: alfieri@uel.br Torque teno sus virus (TTSuV) infection is present in pig herds worldwide. It has been demonstrated that TTSuV might increase the severity of other important viral diseases with economic and public health impacts. At present, there is no information on the age distribution of pigs infected with TTSuV in Brazilian herds. This study evaluated the frequency of TTSuV infection in pigs at different stages of production. Fecal samples (n=190) from pigs at 1 to 24 weeks of age and from breeders at 6 farrow-to-weaning (up to 8 weeks of age) and 9 grower-to-finish (9 weeks of age onwards) farms in the western region of Paraná state, Brazil, were evaluated by PCR. Fragments of the 5’ UTRs of TTSuV1 and/or TTSuVk2 DNAs were identified in 126 (66.3%) of the fecal samples. Significant differences were found with the percentages of positive samples for TTSuV1, TTSuVk2, and mixed infections by both genera between and within the different pig production stages. Fecal samples from the grower-to-finish farms had TTSuV detection rates (90.1%; 64/71) that were significantly (p<0.05) higher than those from the farrow-to-weaning farms (52.1%; 62/119). TTSuV detection was significantly (p<0.05) more frequent in finisher pigs than in the animals from the other stages. The UTR nucleotide sequences in this study presented higher similarities to strains from Norway (96%, TTSuV1), and Argentina and China (97.1%, TTSuVk2). These results suggest that TTSuV infection has spread to pigs of all production stages and that the viral infection rate increases with the age of the animals. In the western region of Paraná state, Brazil, TTSuV1 and TTSuVk2-induced infections were more frequently observed in suckling piglets and finisher pigs, respectively. Phylogenetic analysis pointed out the possibility of different strains of TTSuV1 and TTSuVk2 circulating in pig herds of Brazil.

• orque teno sus virus Iotatorquevirus Kappatorquevirus TTSuV1 TTSuVk2

Abstract in Portuguese:

RESUMO.- Leme R.A., Alfieri A.F. & Alfieri A.A. 2013. Torque teno sus virus (TTSuV) infection at different stages of pig production cycle. [Infecção pelo Torque teno sus virus (TTSuV) em diferentes categorias do ciclo de produção de suínos.] Pesquisa Veterinária Brasileira 33(7):840-846. Laboratório de Virologia Animal, Departamento de Medicina Veterinária Preventiva, Universidade Estadual de Londrina, Rodovia Celso Garcia Cid, Campus Universitário, Cx. Postal 10011, Londrina, PR 86057-970, Brazil E-mail: alfieri@uel.br A infecção pelo Torque teno sus virus (TTSuV) está presente em rebanhos suinícolas em todo o mundo. Tem sido demonstrado que a infecção pelo TTSuV pode aumentar a gravidade de outras importantes doenças virais com impactos econômicos e na saúde pública. Atualmente não há informações sobre a distribuição da infecção pelo TTSuV, de acordo com a faixa etária, em rebanhos suinícolas brasileiros. Este estudo avaliou a frequência da infecção pelo TTSuV nas diferentes categorias de produção de suínos. Amostras fecais (n=190) de suínos com 1 a 24 semanas de idade e de reprodutores provenientes 6 unidades produtoras de leitão (até 8 semanas de idade) e 9 unidades de terminação (9 semanas de idade em diante) da região oeste do Paraná, Brasil, foram avaliadas pela técnica de PCR. Fragmentos da região 5’ UTR do DNA do TTSuV1 e/ou TTSuVk2 foram identificados em 126 (66,3%) amostras fecais. Diferenças significativas foram encontradas em relação às porcentagens de amostras positivas para o TTSuV1, TTSuVk2 e infecção mista por ambos os gêneros inter e intra categorias. Amostras fecais provenientes de unidades de terminação apresentaram taxas de detecção de TTSuV (90.1%; 64/71) significativamente (p<0.05) mais altas do que aquelas provenientes de unidades produtoras de leitão (52.1%; 62/119). A detecção do TTSuV em animais de terminação foi significativamente (p<0.05) mais frequente do que nos suínos de outras categorias. As sequências de nucleotídeos da UTR deste estudo apresentaram maior similaridade com cepas da Noruega (96%, TTSuV1) e Argentina e China (97,1%, TTSuVk2). Estes resultados sugerem que a infecção pelo TTSuV encontra-se disseminada em suínos de todas as categorias de produção e que a taxa da infecção viral aumenta de acordo com a idade dos animais. Na região oeste do estado do Paraná, infecções induzidas pelo TTSuV1 e TTSuVk2 foram mais frequentemente observadas em leitões de maternidade e suínos de terminação, respectivamente. A análise filogenética indicou a possibilidade de diferentes cepas de TTSuV1 e TTSuVk2 circulando em rebanhos suinícolas brasileiros.

Abstract in English:

ABSTRACT.- Dezen S., Otonel R.A.A., Alfieri A.F., Lunardi M. & Alfieri A.A. 2013. [Bovine viral diarrhea virus (BVDV) infection profile in a high production dairy herd with vaccination program against BVDV.] Perfil da infecção pelo vírus da diarreia viral bovina (BVDV) em um rebanho bovino leiteiro de alta produção e com programa de vacinação contra o BVDV. Pesquisa Veterinária Brasileira 33(2):141-147. Laboratório de Virologia Animal, Departamento de Medicina Veterinária Preventiva, Centro de Ciências Agrárias, Universidade Estadual de Londrina, Cx. Postal 6001, Londrina, PR 86051-990, Brazil. E-mail: alfieri@uel.br The profile of bovine viral diarrhea virus (BVDV) infection was studies in a high production dairy herd selected based on a history of reproductive failures and regular vaccination against BVDV. Virus identification was performed by RT-PCR and serological profile was determined by virus-neutralization (VN). Initially, 100% (n=692) of the animals in the herd were monitored for identification of an active infection by RT-PCR in sera. Four months later, all positive animals (n=29) were retested by RT-PCR, along with newly born animals (n=72), or those that had reproductive failures (n=36) in the interval. The RT-PCR assay identified 27 transiently infected animals and three persistently infected (PI). Serology performed only in positive animals in the first RT-PCR and in cows with reproductive failures between the first and second RT-PCR analysis, showed large variation VN antibody titers and seroconversion in most animals. Increases in VN titers were demonstrated, with variation between 3 and 8 log2, indicating virus circulation within the herd. Virus circulation in the vaccinated herd evaluated in this study was likely responsible for reproductive failures observed in cows with low VN titers and for fetal infections. These results demonstrate that control of BVDV infection by regular vaccination in dairy cattle herds with PI animals represents a great challenge for the prophylaxis of this infection.

• BVDV bovine viral diarrhea virus vírus da diarreia viral bovina

Abstract in Portuguese:

RESUMO.- Dezen S., Otonel R.A.A., Alfieri A.F., Lunardi M. & Alfieri A.A. 2013. [Bovine viral diarrhea virus (BVDV) infection profile in a high production dairy herd with vaccination program against BVDV.] Perfil da infecção pelo vírus da diarreia viral bovina (BVDV) em um rebanho bovino leiteiro de alta produção e com programa de vacinação contra o BVDV. Pesquisa Veterinária Brasileira 33(2):141-147. Laboratório de Virologia Animal, Departamento de Medicina Veterinária Preventiva, Centro de Ciências Agrárias, Universidade Estadual de Londrina, Cx. Postal 6001, Londrina, PR 86051-990, Brazil. E-mail: alfieri@uel.br A infecção pelo vírus da diarreia viral bovina (BVDV) foi avaliada em um rebanho bovino leiteiro de alta produção com histórico de problemas reprodutivos e de vacinação regular contra o BVDV. A identificação do vírus foi realizada por RT-PCR em soro sanguíneo e o perfil sorológico por vírus-neutralização. Inicialmente, 100% (n=692) dos animais do rebanho foram avaliados com relação à presença de infecção ativa pelo BVDV por meio da RT-PCR. Quatro meses após, todos os animais positivos (n=29) na primeira avaliação foram avaliados novamente pela RT-PCR, assim como todos os animais que nasceram (n=72) e os que apresentaram problemas reprodutivos (n=36) no intervalo entre a primeira e a segunda colheita de sangue. Os resultados finais do estudo possibilitaram identificar 27 animais transitoriamente infectados e três animais persistentemente infectados (PI). A sorologia, realizada apenas nos animais positivos na primeira avaliação pela RT-PCR e nas vacas que apresentaram problemas reprodutivos entre a primeira e a segunda RT-PCR, demonstrou grande flutuação nos títulos de anticorpos neutralizantes, além de soroconversão na maioria dos animais. Foram identificados aumentos nos títulos de anticorpos neutralizantes que variaram entre 3 e 8 log2, indicando infecção ativa no rebanho. A circulação viral no rebanho avaliado foi responsável pela expressão de sinais clínicos da esfera reprodutiva em animais com baixo título de anticorpos e consequente falha na proteção fetal. Os resultados demonstram que o controle da infecção pelo BVDV apenas por meio da vacinação regular em rebanhos com animais PI pode não ser eficaz na profilaxia dessa virose.

Abstract in English:

ABSTRACT.- Headley S.A., Sousa I.K.F., Minervino A.H.H., Barros I.O., Barrêto Júnior R.A., Alfieri A.F., Ortolani E.L. & Alfieri A.A. 2012. Molecular confirmation of ovine herpesvirus 2-induced malignant catarrhal fever lesions in cattle from Rio Grande do Norte, Brazil. Pesquisa Veterinária Brasileira 32(12):1213-1218. Laboratório de Patologia Veterinária, Faculdade de Medicina Veterinária, Universidade Norte do Paraná, PR 218 Km 1, Cx. Postal 560, Arapongas, PR 86702-670, Brazil. E-mail: headleysa@gmail.com Molecular findings that confirmed the participation of ovine herpesvirus 2 (OHV-2) in the lesions that were consistent with those observed in malignant catarrhal fever of cattle are described. Three mixed-breed cattle from Rio Grande do Norte state demonstrated clinical manifestations that included mucopurulent nasal discharge, corneal opacity and motor incoordination. Routine necropsy examination demonstrated ulcerations and hemorrhage of the oral cavity, corneal opacity, and lymph node enlargement. Significant histopathological findings included widespread necrotizing vasculitis, non-suppurative meningoencephalitis, lymphocytic interstitial nephritis and hepatitis, and thrombosis. PCR assay performed on DNA extracted from kidney and mesenteric lymph node of one animal amplified a product of 423 base pairs corresponding to a target sequence within the ovine herpesvirus 2 (OHV-2) tegument protein gene. Direct sequencing of the PCR products, from extracted DNA of the kidney and mesenteric lymph node of one cow, amplified the partial nucleotide sequences (423 base pairs) of OHV-2 tegument protein gene. Blast analysis confirmed that these sequences have 98-100% identity with similar OHV-2 sequences deposited in GenBank. Phylogenetic analyses, based on the deduced amino acid sequences, demonstrated that the strain of OHV-2 circulating in ruminants from the Brazilian states of Rio Grande do Norte and Minas Gerais are similar to that identified in other geographical locations. These findings confirmed the active participation of OHV-2 in the classical manifestations of sheep associated malignant catarrhal fever.

• Ovine herpesvirus 2 malignant catarrhal fever gammaherpesvirus herpesvírus ovino tipo 2 febre catarral malígna

Abstract in Portuguese:

RESUMO.- Headley S.A., Sousa I.K.F., Minervino A.H.H., Barros I.O., Barrêto Júnior R.A., Alfieri A.F., Ortolani E.L. & Alfieri A.A. 2012. Molecular confirmation of ovine herpesvirus 2-induced malignant catarrhal fever lesions in cattle from Rio Grande do Norte, Brazil. [Diagnóstico molecular de herpesvírus ovino tipo 2 em surto de febre catarral malígna em bovinos do Rio Grande do Norte.] Pesquisa Veterinária Brasileira 32(12):1213-1218. Laboratório de Patologia Veterinária, Faculdade de Medicina Veterinária, Universidade Norte do Paraná, PR 218 Km 1, Cx. Postal 560, Arapongas, PR 86702-670, Brazil. E-mail: headleysa@gmail.com Os achados moleculares confirmaram a participação do herpesvírus ovino tipo 2 (OHV-2) nas lesões observadas em um surto de febre catarral malígna em bovinos. Três bovinos oriundos de propriedade rural de Mossoró, Rio Grande do Norte apresentaram manifestações clínicas, que incluíram secreção nasal mucopurulenta, opacidade da córnea e incoordenação motora. A necropsia revelou ulcerações e hemorragias da cavidade oral, opacidade da córnea e linfonodomegalia. Os achados histopatológicos significativos incluíam vasculite necrosante generalizada, meningoencefalite não supurativa, nefrite intersticial linfocítica, hepatite linfocítica e trombose. A PCR, realizada a partir de DNA extraído do rim e do linfonodo mesentérico de um dos animais, amplificou um produto com 423 pares de base do gene da proteína do tegumento do herpesvírus ovino 2 (OHV-2). O sequenciamento direto dos produtos da PCR e a análise pelo Blast demonstraram que o produto amplificado apresentava 98-100% de identidade com sequências do OHV-2 depositadas no GenBank. As análises filogenéticas, baseadas nas sequências deduzidas de aminoácidos demonstraram que a cepa de OHV-2 circulando em ruminantes nos estados de Rio Grande do Norte e Minas Gerais são semelhantes àquelas identificadas em outras regiões geográficas. Esses achados confirmam a participação ativa de OHV-2 nas manifestações clássicas de febre catarral maligna em ovinos.

Abstract in English:

ABSTRACT.- Dias F.C., Médici K.C., Alexandrino B., Medeiros A.S.R., Alfieri A.A. & Samara S.I. 2010. [Occurrence of persistently infected animals with bovine viral diarrhoea virus in cattle herds from the states of Minas Gerais and São Paulo, Brazil.] Ocorrência de animais persistentemente infectados pelo vírus da diarréia viral bovina em rebanhos bovinos nos Estados de Minas Gerais e São Paulo. Pesquisa Veterinária Brasileira 30(11):933-939. Departamento de Medicina Veterinária Preventiva e Reprodução Animal, Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista, Via de Acesso Prof. Paulo Donato Castellane s/n, Vila Industrial, Jaboticabal, SP 14884-900, Brazil. E-mail: fabiocadi@yahoo.com.br The research on persistently infected (PI) animals with bovine viral diarrhoea virus (BVDV) was conducted in 26 cattle herds, which were not BVDV vaccinated, located in the states of Minas Gerais and São Paulo, Brazil. Using a sampling strategy, five samples of blood were collected from 6 to 12-month-old calves of each herd, and the blood sera were tested by virusneutralization test (VN) to BVDV-1 and BVDV-2. The herds that had at least three out of five samples reacting to one of the genotypes of BVDV and antibody titers greater than 128 were selected to PI animals research. In three of the herds that matched the before-mentioned criteria, paired blood samples were collected from all its individuals considering a collection interval of 30 days. The blood sera of these samples were VN tested against BVDV-1 and BVDV-2. In samples not reacting to at least one of the BVDV genotypes and also in those collected from calves of less than six months of age, virus research was undertaken by reverse transcription polymerase chain reaction (RT-PCR). From the examined herds, two PI animals were detected in paired samples obtained from a herd located in the state of Minas Gerais.

• Bovine viral diarrhoea virus BVDV-1 BVDV-2 vírus da diarreia viral bovina

Abstract in Portuguese:

RESUMO.- Dias F.C., Médici K.C., Alexandrino B., Medeiros A.S.R., Alfieri A.A. & Samara S.I. 2010. [Occurrence of persistently infected animals with bovine viral diarrhoea virus in cattle herds from the states of Minas Gerais and São Paulo, Brazil.] Ocorrência de animais persistentemente infectados pelo vírus da diarréia viral bovina em rebanhos bovinos nos Estados de Minas Gerais e São Paulo. Pesquisa Veterinária Brasileira 30(11):933-939. Departamento de Medicina Veterinária Preventiva e Reprodução Animal, Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista, Via de Acesso Prof. Paulo Donato Castellane s/n, Vila Industrial, Jaboticabal, SP 14884-900, Brazil. E-mail: fabiocadi@yahoo.com.br A pesquisa de animais persistentemente infectados (PI) pelo vírus da diarréia viral bovina (BVDV) foi realizada em 26 rebanhos bovinos, não vacinados contra o BVDV, localizados nos Estados de Minas Gerais e São Paulo, Brasil. Utilizando uma estratégia de amostragem, de cada rebanho foram obtidas cinco amostras de sangue de bezerros, entre 6 e 12 meses de idade, e os soros sanguíneos foram submetidos ao teste de virusneutralização (VN) para o BVDV-1 e o BVDV-2. Os rebanhos que apresentaram pelo menos três das cinco amostras reagentes a um dos genótipos do BVDV, e com títulos de anticorpos superiores a 128, foram selecionados para a pesquisa de animais PI. Em três rebanhos que apresentaram tal condição, foram colhidas amostras pareadas de sangue de todos os bovinos do rebanho, com intervalo de 30 dias entre as colheitas, e o soro sanguíneo foi submetido ao teste de VN para o BVDV-1 e o BVDV-2. Nas amostras não reagentes a pelo menos um dos genótipos do BVDV e naquelas provenientes de bovinos com menos de seis meses de idade, realizou-se a pesquisa do BVDV pela reação em cadeia da polimerase precedida pela transcrição reversa (RT-PCR). Dos rebanhos analisados, foram detectados dois animais PI a partir de amostras obtidas nas colheitas pareadas provenientes de um rebanho localizado no Estado de Minas Gerais.

Abstract in English:

ABSTRACT.- Claus M.P., Lunardi M., Alfieri A.F., Sartori D., Fungaro M.H.P & Alfieri A.A. 2009. Identification of the recently described new type of bovine papillomavirus (BPV-8) in a Brazilian beef cattle herd. Pesquisa Veterinária Brasileira 29(1):25-28. Laboratório de Virologia Animal, Departamento de Medicina Veterinária Preventiva, Centro de Ciências Agrárias, Universidade Estadual de Londrina, Cx. Postal 6001, Campus Universitário, Londrina, PR 86051-990, Brazil. E-mail: alfieri@uel.br Bovine papillomavirus type 8 (BPV-8) was first detected and described in teat warts as well as in healthy teat skin from cattle raised in Japan. The entire viral genome was sequenced in 2007. Additionally, a variant of BPV-8, BPV-8-EB, was also identified from papillomatous lesions of a European bison in Slovakia. In Brazil, despite the relatively common occurrence of BPV infections, the identification and determination of viral types present in cattle is still sporadic. The aim of this study is to report the occurrence of the recently described BPV-8 in Brazil. The virus was identified in a skin warts obtained from a beef cattle herd located in Parana state, southern Brazil. The papilloma had a macular, non-verrucous gross aspect and was located on the dorsal thorax of a cow. Polymerase chain reaction (PCR) was performed using generic primers for partial amplification of L1 gene. The obtained amplicon (480bp) was cloned and two selected clones were sequenced. The nucleotide sequence was compared to existing papillomaviral genomic sequences, identifying the virus as BPV type 8. This study represents the first report of BPV-8 occurrence in Brazil, what suggests its presence among Brazilian cattle.

• Bovine bovine papillomavirus type 8 BPV-8 cutaneous papillomatosis molecular analysis

Abstract in Portuguese:

ABSTRACT.- Claus M.P., Lunardi M., Alfieri A.F., Sartori D., Fungaro M.H.P & Alfieri A.A. 2009. Identification of the recently described new type of bovine papillomavirus (BPV-8) in a Brazilian beef cattle herd. Pesquisa Veterinária Brasileira 29(1):25-28. Laboratório de Virologia Animal, Departamento de Medicina Veterinária Preventiva, Centro de Ciências Agrárias, Universidade Estadual de Londrina, Cx. Postal 6001, Campus Universitário, Londrina, PR 86051-990, Brazil. E-mail: alfieri@uel.br Bovine papillomavirus type 8 (BPV-8) was first detected and described in teat warts as well as in healthy teat skin from cattle raised in Japan. The entire viral genome was sequenced in 2007. Additionally, a variant of BPV-8, BPV-8-EB, was also identified from papillomatous lesions of a European bison in Slovakia. In Brazil, despite the relatively common occurrence of BPV infections, the identification and determination of viral types present in cattle is still sporadic. The aim of this study is to report the occurrence of the recently described BPV-8 in Brazil. The virus was identified in a skin warts obtained from a beef cattle herd located in Parana state, southern Brazil. The papilloma had a macular, non-verrucous gross aspect and was located on the dorsal thorax of a cow. Polymerase chain reaction (PCR) was performed using generic primers for partial amplification of L1 gene. The obtained amplicon (480bp) was cloned and two selected clones were sequenced. The nucleotide sequence was compared to existing papillomaviral genomic sequences, identifying the virus as BPV type 8. This study represents the first report of BPV-8 occurrence in Brazil, what suggests its presence among Brazilian cattle.

Abstract in English:

Abstract.- Zlotowski P., Corrêa A.M.R., Barcellos D.E.S., Cruz C.E.F., Asanome W., Barry A.F., Alfieri A.A. & Driemeier D. 2008. Intestinal lesions in pigs affected with postweaning multisystemic wasting syndrome. Pesquisa Veterinária Brasileira 28(6):313-318. Departamento de Patologia Clinica Veterinária, Faculdade de Medicina Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: davetpat@ufrgs.br Samples of mesenteric lymph nodes and intestines from 79 unthrifty 3- to 5-month-old postweaning pigs, confirmed as naturally affected with postweaning multisystemic wasting syndrome (PMWS), were studied. Pigs originated from 12 farms in southern Brazil and were selected on the basis of clinical signs and/or gross lesions suggestive of enteric disorder. Lymphohistiocytic infiltrates of varying intensity were associated with anti-porcine circovirus type 2 (anti-PCV2) immunostaining (IS) in samples of intestines and mesenteric lymph nodes from all pigs. Although most findings were similar to those described in PCV2-associated enteritis, anti-PCV2 IS in association with depletion of the goblet cell mucin stores (24 pigs), diffuse ileal villous atrophy and fusion (18 pigs), and dilatation of the lymphatic vessels (11 pigs) combined or not with lymphangitis were also observed. PCV2 antigen was immunohistochemically demonstrated in the cytoplasm and nuclei from intralesional epithelial cells, histiocytes, and endothelial-like cells in intestinal tissues. Together these findings imply an association with PCV2. The presence of co-infections by Lawsonia intracellularis, Brachyspira spp., Mycobacterium spp., Salmonella spp., rotavirus, parvovirus, coronavirus and enteric calicivirus with PCV2 in the intestinal lesions was investigated.

• Circovirus co-infections enteric lesions PCV2 PMWS

Abstract in Portuguese:

Abstract.- Zlotowski P., Corrêa A.M.R., Barcellos D.E.S., Cruz C.E.F., Asanome W., Barry A.F., Alfieri A.A. & Driemeier D. 2008. Intestinal lesions in pigs affected with postweaning multisystemic wasting syndrome. Pesquisa Veterinária Brasileira 28(6):313-318. Departamento de Patologia Clinica Veterinária, Faculdade de Medicina Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: davetpat@ufrgs.br Samples of mesenteric lymph nodes and intestines from 79 unthrifty 3- to 5-month-old postweaning pigs, confirmed as naturally affected with postweaning multisystemic wasting syndrome (PMWS), were studied. Pigs originated from 12 farms in southern Brazil and were selected on the basis of clinical signs and/or gross lesions suggestive of enteric disorder. Lymphohistiocytic infiltrates of varying intensity were associated with anti-porcine circovirus type 2 (anti-PCV2) immunostaining (IS) in samples of intestines and mesenteric lymph nodes from all pigs. Although most findings were similar to those described in PCV2-associated enteritis, anti-PCV2 IS in association with depletion of the goblet cell mucin stores (24 pigs), diffuse ileal villous atrophy and fusion (18 pigs), and dilatation of the lymphatic vessels (11 pigs) combined or not with lymphangitis were also observed. PCV2 antigen was immunohistochemically demonstrated in the cytoplasm and nuclei from intralesional epithelial cells, histiocytes, and endothelial-like cells in intestinal tissues. Together these findings imply an association with PCV2. The presence of co-infections by Lawsonia intracellularis, Brachyspira spp., Mycobacterium spp., Salmonella spp., rotavirus, parvovirus, coronavirus and enteric calicivirus with PCV2 in the intestinal lesions was investigated.

Abstract in English:

Abstract.- Barry A.F, Alfieri A.F. & Alfieri A.A. 2008. Detection and phylogenetic analysis of porcine enteric calicivirus, genetically related to the Cowden strain of sapovirus genogroup III, in Brazilian swine herds. Pesquisa Veterinária Brasileira 28(1):82-86. Laboratório de Virologia Animal, Departamento de Medicina Veterinária Preventiva, Centro de Ciências Agrárias, Universidade Estadual de Londrina, Campus Universitário, Londrina, PR 86051-990, Brazil. E-mail: alinebarry@uol.com.br Sapovirus of the Caliciviridae family is an important agent of acute gastroenteritis in children and piglets. The Sapovirus genus is divided into seven genogroups (G), and strains from the GIII, GVI and GVII are associated with infections in swine. Despite the high prevalence in some countries, there are no studies related to the presence of porcine enteric sapovirus infections in piglets in Brazil. In the present study, 18 fecal specimens from piglets up to 28 days were examined to determine the presence of sapovirus genome by RT-PCR assay, using primers designed to amplify a 331 bp segment of the RNA polymerase gene. In 44.4% (8/18) of fecal samples, an amplified DNA fragment was obtained. One of these fragments was sequenced and submitted to molecular and phylogenetic analysis. This analysis revealed high similarity, with nucleotides (87%) and amino acids (97.8%), to the Cowden strain, the GIII prototype of porcine enteric calicivirus. This is the first description of sapovirus in Brazilian swine herds.

• Swine; piglets diarrhea calicivirus Sapovirus

Abstract in Portuguese:

Abstract.- Barry A.F, Alfieri A.F. & Alfieri A.A. 2008. Detection and phylogenetic analysis of porcine enteric calicivirus, genetically related to the Cowden strain of sapovirus genogroup III, in Brazilian swine herds. Pesquisa Veterinária Brasileira 28(1):82-86. Laboratório de Virologia Animal, Departamento de Medicina Veterinária Preventiva, Centro de Ciências Agrárias, Universidade Estadual de Londrina, Campus Universitário, Londrina, PR 86051-990, Brazil. E-mail: alinebarry@uol.com.br Sapovirus of the Caliciviridae family is an important agent of acute gastroenteritis in children and piglets. The Sapovirus genus is divided into seven genogroups (G), and strains from the GIII, GVI and GVII are associated with infections in swine. Despite the high prevalence in some countries, there are no studies related to the presence of porcine enteric sapovirus infections in piglets in Brazil. In the present study, 18 fecal specimens from piglets up to 28 days were examined to determine the presence of sapovirus genome by RT-PCR assay, using primers designed to amplify a 331 bp segment of the RNA polymerase gene. In 44.4% (8/18) of fecal samples, an amplified DNA fragment was obtained. One of these fragments was sequenced and submitted to molecular and phylogenetic analysis. This analysis revealed high similarity, with nucleotides (87%) and amino acids (97.8%), to the Cowden strain, the GIII prototype of porcine enteric calicivirus. This is the first description of sapovirus in Brazilian swine herds.

Abstract in English:

ABSTRACT.- Oliveira Filho J.P., Silva D.P.G., Pacheco M.D., Mascarini L.M., Marcio Garcia Ribeiro, Alfieri A.A., Alfieri A.F., Stipp D.T., Barros B.J.P. & Borges A.S. 2007. [Diarrhea in Nelore calves: Clinical and etiologic study.] Diarréia em bezerros da raça Nelore criados extensivamente: estudo clínico e etiológico. Pesquisa Veterinária Brasileira 27(10):419-424. Departamento de Clínica Veterinária, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual Paulista, Campus de Botucatu s/n, Distrito de Rubião Júnior, Botucatu, SP 18618000, Brazil. E-mail: zep.filho@hotmail.com Diarrhea is considered as one of the main causes of morbidity and mortality in neonates calves. Fecal samples from 100 diarrheic and 30 non-diarrheic (control group) Nelore calves less than 9 weeks old were collected for Salmonella spp., Escherichia coli, rotavirus, coronavirus, Cryptosporidium spp., and for helminth eggs investigation. Enteropathogens were detected in 79.0% diarrheic samples and 70.0% non-diarrheic samples. Among diarrheic calves, Escherichia coli (69.0%) was the most common agent found, following by Cryptosporidium spp. (30.0%), coronavirus (16.0%), and rotavirus (11.0%). In the control group, E. coli, Cryptosporidium spp. and coronavirus were detected in 66.7%, 10.0% and 3.3% of the samples, respectively. Salmonella spp. and strongylids were not found in any of the calves from either group. The K99 fimbrial only was detected in E. coli strains from diarrheic calves (5.8%). Enrofloxacin, norfloxacin, and gentamicin were the most effective among the antimicrobials tested. The weight of 210-day-old calves did not show statistic differences between diarrheic and non-diarrheic calves.

• Beef calves Nelore diarrhea enteropathogens etiology

Abstract in Portuguese:

ABSTRACT.- Oliveira Filho J.P., Silva D.P.G., Pacheco M.D., Mascarini L.M., Marcio Garcia Ribeiro, Alfieri A.A., Alfieri A.F., Stipp D.T., Barros B.J.P. & Borges A.S. 2007. [Diarrhea in Nelore calves: Clinical and etiologic study.] Diarréia em bezerros da raça Nelore criados extensivamente: estudo clínico e etiológico. Pesquisa Veterinária Brasileira 27(10):419-424. Departamento de Clínica Veterinária, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual Paulista, Campus de Botucatu s/n, Distrito de Rubião Júnior, Botucatu, SP 18618000, Brazil. E-mail: zep.filho@hotmail.com Diarrhea is considered as one of the main causes of morbidity and mortality in neonates calves. Fecal samples from 100 diarrheic and 30 non-diarrheic (control group) Nelore calves less than 9 weeks old were collected for Salmonella spp., Escherichia coli, rotavirus, coronavirus, Cryptosporidium spp., and for helminth eggs investigation. Enteropathogens were detected in 79.0% diarrheic samples and 70.0% non-diarrheic samples. Among diarrheic calves, Escherichia coli (69.0%) was the most common agent found, following by Cryptosporidium spp. (30.0%), coronavirus (16.0%), and rotavirus (11.0%). In the control group, E. coli, Cryptosporidium spp. and coronavirus were detected in 66.7%, 10.0% and 3.3% of the samples, respectively. Salmonella spp. and strongylids were not found in any of the calves from either group. The K99 fimbrial only was detected in E. coli strains from diarrheic calves (5.8%). Enrofloxacin, norfloxacin, and gentamicin were the most effective among the antimicrobials tested. The weight of 210-day-old calves did not show statistic differences between diarrheic and non-diarrheic calves.