PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- De Alcântara B.K., Alfieri A.A., Headley S.A., Rodrigues W.B., Otonel R.A.A., Lunardi M. & Alfieri A.F. 2015. Molecular characterization of bovine Deltapapillomavirus (BPV1, 2, and 13) DNA in equine sarcoids. Pesquisa Veterinária Brasileira 35(5):431-436. 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 Sarcoids are fibroblastic lesions, which are considered as the most common skin tumors of horses; spontaneous regression rarely occurs. The bovine papillomavirus (BPV) types 1 and 2 may be involved in the pathogenesis of sarcoids, and probably the recently described BPV type (BPV13) might be associated with the pathogenesis of this lesion. This study characterized the DNA of BPVs in sarcoids from 15 horses from Brazil by analyzing 20 cutaneous lesions (12 recently collected; 8 from formalin-fixed paraffin-embedded (FFPE) tissues). Histopathology confirmed the proliferative lesions as sarcoids. Three PCRs were performed to amplify papillomavirus (PV) DNA. For screening, the primers IFNR2/IDNT2 were used to amplify a fragment of the PV L1 ORF. The second primer set was complementary to a common sequence of the E5L2 genomic region of BPV1, 2, and 13. The third primer pair (FAP59/FAP64) targeted a fragment of the PVs L1 ORF. The screening and E5L2 PCRs yielded amplicons in all samples evaluated. The FAP amplicons identified BPV1, 2, and 13 only from fresh tissue samples. The phylogenetic analyses of E5L2 resulted in the identification of BPV1, 2, and 13 in 14 (70%), 2 (10%), and 4 (20%) sarcoids, respectively. Two horses demonstrated multiple lesions: the sarcoids of one of these contained only BPV1 DNA and those of the other contained three types of bovine Deltapapillomavirus (BPV1, 2, and 13). This study confirmed the presence of BPV1, 2, and 13 DNA in equine sarcoids. Moreover, these findings represent the first description of three types of BPV diagnosed in the same horse, as well as the first confirmation of BPV1 and 2 in horses from Brazil.

• Papillomavirus Bovine Deltapapillomavirus skin tumor Papilomavírus Deltapapillomavirus bovino tumor de pele

Abstract in Portuguese:

RESUMO.- De Alcântara B.K., Alfieri A.A., Headley S.A., Rodrigues W.B., Otonel R.A.A., Lunardi M. & Alfieri A.F. 2015. Molecular characterization of bovine Deltapapillomavirus (BPV1, 2, and 13) DNA in equine sarcoids. [Caracterização molecular de DNA de Deltapapillomavirus bovino (BPV1, 2 e 13) em sarcoides equinos.] Pesquisa Veterinária Brasileira 35(5):431-436. 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 Sarcoides são tumores fibroblásticos, considerados os tumores de pele mais comuns em pele de equinos e que raramente apresentam regressão espontânea. Papilomavírus bovino (BPV) tipos 1 e 2 são relacionados com a patogenia do sarcoide e, provavelmente, o BPV tipo 13 (BPV13), recentemente descrito, também pode estar associado com a formação dessa lesão. Neste estudo, 20 amostras de lesões cutâneas, sendo 12 constituídas por tecidos frescos e 8 amostras de tecido fixado em formalina e embebido em parafina, provenientes de 15 cavalos foram utilizadas para a identificação do DNA de BPV. A análise histopatológica (HE) confirmou todas as lesões como sarcoide. Para a amplificação do DNA de papilomavírus (PV) foram realizadas três reações de PCR. Como triagem, os primers IFNR2/IDNT2 foram utilizados para amplificar um fragmento da ORF L1 do PV. O segundo par de primers utilizado é complementar a sequência dos genes E5 e L2 de BPVs 1, 2 e 13. O terceiro par de primers (FAP59/FAP64) utilizado tem o gene L1 como alvo. A primeira e a segunda PCRs permitiram amplificar produtos em todas as amostras avaliadas. Entretanto, na terceira reação, na qual foram utilizados os primers FAP, foi possível amplificar produtos com tamanho molecular esperado somente nas amostras constituídas por tecidos frescos. O sequenciamento de nucleotídeos e as análises filogenéticas realizadas nos fragmentos E5L2 resultaram na identificação de BPV1, 2 e 13 em 14 (70%), 2 (10%) e em 4 (20%) amostras de sarcoides, respectivamente. As amostras de sarcoides de um dos animais continha somente o DNA de BPV1. Entretanto, nas amostras provenientes do segundo cavalo foi possível identificar o DNA de três tipos de Deltapapillomavirus bovino (BPV1, 2 e 13) em lesões distintas. Este estudo ratifica a presença do DNA de BPV1, 2 e 13 em lesões de sarcoides em equinos, além de identificar três tipos de BPVs em um mesmo animal e descrever pela primeira vez no Brasil a presença de BPV1 e 2 nesse tipo de lesão.

Abstract in English:

ABSTRACT.- Masuda E.K., Kommers G.D., Rosa F.B., Barros C.S.L., Fighera R.A. & Piazer J.V.M. 2011. [Relationship between lymphopenia and the persistence of alimentary papillomatosis in cattle chronically and spontaneously poisoned by bracken fern (Pteridium aquilinum).] Relação entre a linfopenia e a persistência da papilomatose alimentar em bovinos intoxicados crônica e espontaneamente por samambaia (Pteridium aquilinum). Pesquisa Veterinária Brasileira 31(5):383-388. Laboratório de Patologia Veterinária, Departamento de Patologia, Centro de Ciências da Saúde, Universidade Federal de Santa Maria, Avenida Roraima 1000, Santa Maria, RS 97105-900, Brazil. E-mail:glaukommers@yahoo.com Bovine papillomavirus type-4 (BPV-4) is responsible for the development of papillomas in the upper digestive tract (UDT) of cattle. The necessary steps for the development, growth and regression of papillomas are intimately related with the immunological status of the animal. The consumption of bracken fern (Pteridium aquilinum) has been pointed out as the major factor involved in the persistence of BPV-4 infection of the UDT. The theory involving the relationship between alimentary papillomas and the formation of squamous cell carcinomas (SCCs) suggests the persistence of alimentary papillomatosis through a chronic immunosupressive lymphopenic state induced by bracken fern. The papillomas would serve then as a development site of SCCs through the interaction of BPV-4 oncoproteins and bracken carcinogens. The objective of this study was to evaluate the relationship between lymphopenia and alimentary papillomatosis in spontaneous cases of chronic bracken fern poisoning of cattle. The age, intensity of papillomatosis and the leukon were evaluated of 40 cattle with SCCs of the UDT. Three animals had leukopenia and one had neutrophilia. The mean lymphocyte count (and standart deviation) of mild, moderate and severe papillomatosis was 5,395(±1,696)/mm3, 4,560(±1,561)/mm3 and 5,007(±1,786)/mm3, respectively. There were no statistically significant differences between the intensity of alimentary papillomasis, age and the lymphocyte count. Immunosupression by lymphopenia was a sporadic finding (three cases) in the present study. The results demonstrate that the persistence of alimentary papilomatosis in spontaneous cases of bracken fern-related SCCs of the UDT is not related to lymphopenia and it may be related to other immunological factors.

• Pteridium aquilinum papillomatosis bovine papillomavirus type 4 plant poisoning diseases of cattle Plantas tóxicas papilomatose papilomavírus bovino tipo 4 intoxicação por planta

Abstract in Portuguese:

RESUMO.- Masuda E.K., Kommers G.D., Rosa F.B., Barros C.S.L., Fighera R.A. & Piazer J.V.M. 2011. [Relationship between lymphopenia and the persistence of alimentary papillomatosis in cattle chronically and spontaneously poisoned by bracken fern (Pteridium aquilinum).] Relação entre a linfopenia e a persistência da papilomatose alimentar em bovinos intoxicados crônica e espontaneamente por samambaia (Pteridium aquilinum). Pesquisa Veterinária Brasileira 31(5):383-388. Laboratório de Patologia Veterinária, Departamento de Patologia, Centro de Ciências da Saúde, Universidade Federal de Santa Maria, Avenida Roraima 1000, Santa Maria, RS 97105-900, Brazil. E-mail:glaukommers@yahoo.com Papilomavírus bovino tipo 4 (BPV-4) é responsável pelo desenvolvimento de papilomas no trato alimentar superior (TAS) de bovinos. Os passos necessários para o crescimento, o desenvolvimento e a regressão dos papilomas estão intimamente relacionados com o estado imunológico do animal. A ingestão de samambaia (Pteridium aquilinum) tem sido relacionada como o principal fator envolvido na persistência da infecção pelo BPV-4 no TAS. A teoria que estabelece a relação entre papilomatose alimentar e a formação de CCEs sugere a produção de um estado imunossupressivo crônico pela planta, permitindo a persistência dos papilomas no TAS. Os papilomas serviriam então como sítios de desenvolvimento dos CCEs através da interação entre as proteínas do BPV-4 e os carcinógenos da samambaia. O objetivo deste estudo foi avaliar a relação entre a quantidade de linfócitos circulantes e a papilomatose alimentar em casos de intoxicação espontânea crônica por P. aquilinum em bovinos com CCE no TAS. Quarenta bovinos com CCEs no TAS foram avaliados quanto à idade, à intensidade da papilomatose alimentar no TAS e ao leucograma. Três bovinos tinham leucopenia e um apresentava neutrofilia. A média de linfócitos foi de 5.395 (±1.696) na papilomatose leve, 4.560 (±1.561) na moderada e 5.007 (±1.786) na acentuada. Não houve diferença estatisticamente significativa entre o grau de papilomatose, a idade e a quantidade de linfócitos circulantes. Imunossupressão por linfopenia foi um achado esporádico (três casos) neste estudo. Os resultados indicam que a persistência da papilomatose alimentar em casos espontâneos de intoxicação crônica por samambaia em bovinos não tem relação com a quantidade de linfócitos circulantes e que talvez esteja relacionada a outros fatores imunológicos.

Abstract in English:

ABSTRACT.- Claus M.P., Vivian D., Lunardi M., Alfieri A.F. & Alfieri A.A. 2007. [Phylogenetic analysis of bovine papillomavirus associated with skin warts in cattle herds from the state of Paraná.] Análise filogenética do papilomavírus bovino associado a lesões cutâneas em rebanhos do Estado do Paraná. Pesquisa Veterinária Brasileira 27(7):314-318. 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 (BPV) infection causes hyperplastic lesions in the cutaneous epithelium of cattle. Six types of BPV were classified in two sub-groups, being correlated to the anatomical regions of the infection and morphologic characteristics of the lesions. The present study was carried out to identify the types of BPV present in skin warts of cattle from the state of Paraná, Brazil. The generic primers FAP59 and FAP64 were used for amplification of a 478 bp fragment of BPV L1 gene in nine cutaneous papilloma samples obtained from six animals in four herds. In all papillomas examined, a product with the expected molecular size was amplified. Phylogenetic analysis of the PCR products identified BPV-2 in three samples, BPV-1 in one, and BPV-6 in five papillomas. BPV-6 was detected in cutaneous papillomas of the teat and in other body parts as well. In one animal, from which more than one sample was collected, a concomitant infection by BPV-1 and BPV-2 was identified. The five positive BPV-6 samples showed a nucleotide identity of 100% with the sequence of the reference strain available in GenBank. However, differences among BPV-2 and BPV-1 Brazilian samples and the respective reference sequences deposited in GenBank were observed. Molecular comparison of the two BPV-2 strains identified showed the involvement of two viral variants. This study revealed the diversity of BPV types circulating in the state of Paraná.

• Bovine papillomavirus cutaneous papillomatosis phylogeny polymerase chain reaction

Abstract in Portuguese:

ABSTRACT.- Claus M.P., Vivian D., Lunardi M., Alfieri A.F. & Alfieri A.A. 2007. [Phylogenetic analysis of bovine papillomavirus associated with skin warts in cattle herds from the state of Paraná.] Análise filogenética do papilomavírus bovino associado a lesões cutâneas em rebanhos do Estado do Paraná. Pesquisa Veterinária Brasileira 27(7):314-318. 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 (BPV) infection causes hyperplastic lesions in the cutaneous epithelium of cattle. Six types of BPV were classified in two sub-groups, being correlated to the anatomical regions of the infection and morphologic characteristics of the lesions. The present study was carried out to identify the types of BPV present in skin warts of cattle from the state of Paraná, Brazil. The generic primers FAP59 and FAP64 were used for amplification of a 478 bp fragment of BPV L1 gene in nine cutaneous papilloma samples obtained from six animals in four herds. In all papillomas examined, a product with the expected molecular size was amplified. Phylogenetic analysis of the PCR products identified BPV-2 in three samples, BPV-1 in one, and BPV-6 in five papillomas. BPV-6 was detected in cutaneous papillomas of the teat and in other body parts as well. In one animal, from which more than one sample was collected, a concomitant infection by BPV-1 and BPV-2 was identified. The five positive BPV-6 samples showed a nucleotide identity of 100% with the sequence of the reference strain available in GenBank. However, differences among BPV-2 and BPV-1 Brazilian samples and the respective reference sequences deposited in GenBank were observed. Molecular comparison of the two BPV-2 strains identified showed the involvement of two viral variants. This study revealed the diversity of BPV types circulating in the state of Paraná.