PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Neves S.M.N., Gabardo M.P. & Guedes R.M.C. 2017. [Fluorescence in situ hybridization for diagnosis of Brachyspira hyodysenteriae e B. pilosicoli in swine.] Hibridização in situ fluorescente para diagnóstico de Brachyspira hyodysenteriae e B. pilosicoli em suínos. Pesquisa Veterinária Brasileira 37(10):1101-1107. Departamento de Clínica e Cirurgia, Escola de Veterinária, Universidade Federal de Minas Gerais, Av. Antônio Carlos 6627, Pampulha, Belo Horizonte, MG 31270-901, Brazil. E-mail: guedesufmg@gmail.com Growing and finishing pigs are affected by pathogenic spirochetes of the genus Brachyspira sp., which cause a significant economic impact due to direct and indirect losses. Thus, efficient diagnosis of these species enables better technical intervention to prevent or treat diseases. This study aimed to evaluate the fluorescent in situ hybridization (FISH) for the diagnosis of B. hyodysenteriae and B. pilosicoli in histopathologic fragments of pig’s intestine and compare it to the duplex PCR. Thirty-four samples collected from pigs positive for these species in at least one of the tests were used in the study. Out of the 34 analyzed intestine samples, 28 (82.35%) were positive by PCR and FISH. Among the 29 B. hyodysenteriae positive samples, 23 (79.3%) were positive by PCR and 21 (72.4%) by FISH. There was no statistical difference among the detection rate of the used tests. Based on the fact this technique can be performed in formalin fixed tissue samples, it is practical, fast and allows the association of labeling a specific agent with histological lesions, FISH has become an alternative diagnostic method for Brachyspira hyodysenteriae and B. pilosicoli.

• Hybridization Brachyspira hyodysenteriae Brachyspira pilosicoli swine spirochaetes dysentery diarrhea suínos espiroquetas disenteria suína colite espiroquetal

Abstract in Portuguese:

RESUMO.- Neves S.M.N., Gabardo M.P. & Guedes R.M.C. 2017. [Fluorescence in situ hybridization for diagnosis of Brachyspira hyodysenteriae e B. pilosicoli in swine.] Hibridização in situ fluorescente para diagnóstico de Brachyspira hyodysenteriae e B. pilosicoli em suínos. Pesquisa Veterinária Brasileira 37(10):1101-1107. Departamento de Clínica e Cirurgia, Escola de Veterinária, Universidade Federal de Minas Gerais, Av. Antônio Carlos 6627, Pampulha, Belo Horizonte, MG 31270-901, Brazil. E-mail: guedesufmg@gmail.com Disenteria Suína e Colite Espiroquetal são duas enfermidades importantes em suínos causados pela Brachyspira hyodysenteriae e Brachyspira pilosicoli, respectivamente. O diagnóstico eficaz dessas espécies é extremamente importante para a adoção de estratégias adequadas para o controle. Propõe-se avaliar a técnica de hibridização in situ de fluorescência (FISH) para detecção de B. hyodysenteriae e B. pilosicoli em fragmentos histopatológicos de intestino de suínos e compará-la ao PCR duplex. Foram analisadas amostras de fezes e intestinos de suínos de terminação com histórico de diarreia pelas técnicas de reação em cadeia da polimerase duplex (dPCR), hibridização in situ fluorescente (FISH) para diagnóstico dessas bactérias. Foram utilizadas 34 amostras de intestino de suínos de campo positivos para alguma das duas espécies de Brachyspira sp. nos testes de FISH ou PCR. Das 34 amostras analisadas, foram detectadas 28 (82,35%) positivas na PCR e no FISH. Dentre as 29 amostras positivas para B. hyodysenteriae, 23 (79,3%) foram positivas à PCR e 21 (72,4%) no FISH. Os resultados de FISH e PCR não diferiram estatisticamente entre si. Baseado no fato dessa técnica poder ser realizada em tecidos formolizados, ser prática, rápida e associar a marcação especifica do agente com lesões histológicas, o FISH demonstrou ser mais uma alternativa no diagnóstico de Brachyspira hyodysenteriae e B. pilosicoli.

Abstract in English:

ABSTRACT.- Daniel A.G.S., Sato J.P.H., Gabardo M.P., Resende T.P., Barcellos D.E.S.N., Pereira C.E.R. Vannucci F.A. & Guedes R.M.C. 2017. Minimum inhibitory concentration of Brazilian Brachyspira hyodysenteriae strains. Pesquisa Veterinária Brasileira 37(4):331-338. Departamento de Clínica e Cirurgia Veterinária, Escola de Veterinária, Universidade Federal de Minas Gerais, Avenida Pres. Antônio Carlos 6627, Pampulha, Belo Horizonte, MG 31270-901, Brazil. E-mail: guedesufmg@gmail.com The objectives of this study were to characterize Brachyspira hyodysenteriae isolates and to evaluate the antimicrobial susceptibility patterns of strains obtained from pigs in Brazil based on the minimal inhibitory concentration test (MIC). The MIC was performed for 22 B. hyodysenteriae isolates obtained from 2011 to 2013 using the following antimicrobial drugs: tylosin, tiamulin, valnemulin, doxycycline, lincomycin and tylvalosin. Outbreaks of swine dysentery were diagnosed based on clinical presentation, bacterial isolation, gross and microscopic lesions, duplex PCR for B. hyodysenteriae and B. pilosicoli and nox gene sequencing. All obtained MIC values were consistently higher or equal to the microbiological cut-off described in the literature. The MIC 90 values for the tested drugs were 8µg/ml for doxycycline, >4µg/ml for valnemulin, 8µg/ml for tiamulin, 32µg/ml for tylvalosin, >64µg/ml for lincomycin and >128µg/ml for tylosin. These results largely corroborate those reported in the literature. Tiamulin, doxycycline and tylvalosin showed the lowest MIC results. All of the samples subjected to phylogenetic analysis based on the nox gene sequence exhibited similar results, showing 100% identity to B. hyodysenteriae. This is the first study describing the MIC pattern of B. hyodysenteriae isolated in Brazil.

• Brachyspira hyodysenteriae swine dysentery diarrhea antimicrobial sensitivity nox gene disenteria suína sensibilidade antimicrobiana gene nox

Abstract in Portuguese:

RESUMO.- Daniel A.G.S., Sato J.P.H., Gabardo M.P., Resende T.P., Barcellos D.E.S.N., Pereira C.E.R. Vannucci F.A. & Guedes R.M.C. 2017. Minimum inhibitory concentration of Brazilian Brachyspira hyodysenteriae strains. [Concentração inibitória mínima de cepas Brachyspira hyodysenteriae brasileiras.] Pesquisa Veterinária Brasileira 37(4):331-338. Departamento de Clínica e Cirurgia Veterinária, Escola de Veterinária, Universidade Federal de Minas Gerais, Avenida Pres. Antônio Carlos 6627, Pampulha, Belo Horizonte, MG 31270-901, Brazil. E-mail: guedesufmg@gmail.com Os objetivos deste trabalho foram a caracterização de isolados de Brachyspira hyodysenteriae e avaliar os padrões de sensibilidade antimicrobiana de isolados obtidos a partir de suínos no Brasil com base no teste de concentração inibitória mínima (MIC). A MIC foi realizada em 22 isolados de B. hyodysenteriae obtidos entre 2011 a 2013 usando os seguintes antimicrobianos: tilosina, tiamulina, valnemulina, doxiciclina, lincomicina e tilvalosina. Surtos de disenteria suína foram diagnosticados com base na apresentação clínica, isolamento bacteriano, lesões macroscópicas e microscópicas, PCR duplex para B. hyodysenteriae e B. pilosicoli e sequenciamento do gene nox. Todos os valores de MIC obtidos foram consistentemente mais elevados ou igual ao ponto de corte microbiológica descrito na literatura. Os valores de MIC 90 para os fármacos testados foram de 8 µg / mL para a doxiciclina, > 4 µg/ml de valnemulina, 8 µg / mL para a tiamulina, 32 µg / ml para tilvalosina, > 64 µg / ml para a lincomicina e > 128 μg / ml de tilosina. Estes resultados corroboram em grande parte com os relatados na literatura. Tiamulina, doxiciclina e tilvalosina apresentaram os menores resultados de MIC. Todas as amostras submetidas à análise filogenética com base na sequência do gene nox exibiram resultados semelhantes, indicando 100% de identidade com B. hyodysenteriae. Este é o primeiro estudo que descreve o padrão MIC de B. hyodysenteriae isoladas no Brasil.

Abstract in English:

Abstract.- Pavarini S.P., Bezerra Junior P.S., Santos A.S., Pescador C.A. & Driemeier D. 2008. [Clinic pathological features during a winter dysentery outbreak in cattle in Rio Grande do Sul.] Achados clínico patológicos durante um surto de disenteria de inverno em bovinos no Rio Grande do Sul. Pesquisa Veterinária Brasileira 28(12):611- 616. Departamento de Patologia Clínica Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: davetpat@ufrgs.br The report describes a winter dysentery outbreak that affected 10 dairy cows from a ranch located in the county of Viamão, Rio Grande do Sul. The most significant clinical sign was profuse and watery diarrhea, which ranged from a greenish to a brownish coloration and from occasional blood streaks with mucus to a bloody diarrhea. Most cases persisted for 5 days and also included depression, drastic decrease in milk production and in food consumption. Only one of the affected animals died. The necropsy revealed pale mucosa and sanguineous content with high quantity of blood clots, particularly within the spiral colon, and petechiae on the colonic mucosa. Histopathological lesions were predominant in the spiral colon, and consisted of a high number of dilated crypts without epithelium or with the replacement of a pavement epithelium with occasional immature cuboidal cells, which sometimes showed enlarged nucleus and proeminent nucleolus. Some crypts were filled with epithelial desquamation and polymorphnuclear cells. Bovine coronavirus (8F2) monoclonal antibody was used for immunohistochemistry on sections of the spiral colon that showed positive reactions in the cytoplasm of the infected crypt epithelium, sloughed necrotic cells, and within macrophages in the lamina propria of both.

• Winter dysentery diarrhea spiral colon bovine coronavirus

Abstract in Portuguese:

Abstract.- Pavarini S.P., Bezerra Junior P.S., Santos A.S., Pescador C.A. & Driemeier D. 2008. [Clinic pathological features during a winter dysentery outbreak in cattle in Rio Grande do Sul.] Achados clínico patológicos durante um surto de disenteria de inverno em bovinos no Rio Grande do Sul. Pesquisa Veterinária Brasileira 28(12):611- 616. Departamento de Patologia Clínica Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: davetpat@ufrgs.br The report describes a winter dysentery outbreak that affected 10 dairy cows from a ranch located in the county of Viamão, Rio Grande do Sul. The most significant clinical sign was profuse and watery diarrhea, which ranged from a greenish to a brownish coloration and from occasional blood streaks with mucus to a bloody diarrhea. Most cases persisted for 5 days and also included depression, drastic decrease in milk production and in food consumption. Only one of the affected animals died. The necropsy revealed pale mucosa and sanguineous content with high quantity of blood clots, particularly within the spiral colon, and petechiae on the colonic mucosa. Histopathological lesions were predominant in the spiral colon, and consisted of a high number of dilated crypts without epithelium or with the replacement of a pavement epithelium with occasional immature cuboidal cells, which sometimes showed enlarged nucleus and proeminent nucleolus. Some crypts were filled with epithelial desquamation and polymorphnuclear cells. Bovine coronavirus (8F2) monoclonal antibody was used for immunohistochemistry on sections of the spiral colon that showed positive reactions in the cytoplasm of the infected crypt epithelium, sloughed necrotic cells, and within macrophages in the lamina propria of both.

Abstract in English:

ABSTRACT.- Brandão P.E., Laura Y. B. Villarreal L.Y.B., F.Gregori F., Souza S.L.P., Lopes M.A.E., Gomes C.R., Sforsin A.J., Sanches A.A., Rosales C.A.R., Richtzenhain L.J., Ferreira A.J.P. & Jerez J.A. 2007. On the etiology of an outbreak of winter dysentery in dairy cows in Brazil. Pesquisa Veterinária Brasileira 27(10):398-402. Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, São Paulo, SP 05508-270, Brazil. E-mail: paulo7926@yahoo.com Winter dysentery (WD) is a seasonal infectious disease described worldwide that causes a marked decrease in milk production in dairy cows. In the Northern hemisphere, where the disease is classically recognized, bovine coronavirus (BCoV) has been assigned as a major etiologic agent of the disease. Nonetheless, in the Southern hemisphere, an in-deep etiological survey on WD cases had not been carried out. This study aimed to survey for BCoV by nested-RT-PCR, rotavirus by polyacrylamide gel electrophoresis (PAGE) and ELISA, bacteria by classical bacteriological methods and PCR for virulence factors and parasites by sugar flotation test on fecal samples of 21 cows from a farm during an outbreak of WD in São Paulo state, Southeastern Brazil. BCoV was detected in all 21 samples, while rotavirus was detected in two symptomatic cows. Escherichia coli, Yersinia intermedia, Providencia rustigianii Proteus penneri, Klebsiella terrigena and Enterobacter aglomerans were detected in samples from both asymptomatic and healthy cows in different associations. The study of E. coli virulence factors revealed that the strains isolated were all apathogenic. Cysts of Eimeria sp. and eggs of Strongyloidea were detected at low numbers in four of the symptomatic cows, with one co-infestation. These results suggest BCoV as the main etiologic agent of the cases of WD in Brazil, a conclusion that, with the clinical and epidemiological patterns of the disease studied herein, match those already described elsewhere. These findings give basis to the development of preventive measures and contribute to the understanding of the etiology of WD.

• Bovine coronavirus etiology winter dysentery

Abstract in Portuguese:

ABSTRACT.- Brandão P.E., Laura Y. B. Villarreal L.Y.B., F.Gregori F., Souza S.L.P., Lopes M.A.E., Gomes C.R., Sforsin A.J., Sanches A.A., Rosales C.A.R., Richtzenhain L.J., Ferreira A.J.P. & Jerez J.A. 2007. On the etiology of an outbreak of winter dysentery in dairy cows in Brazil. Pesquisa Veterinária Brasileira 27(10):398-402. Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, São Paulo, SP 05508-270, Brazil. E-mail: paulo7926@yahoo.com Winter dysentery (WD) is a seasonal infectious disease described worldwide that causes a marked decrease in milk production in dairy cows. In the Northern hemisphere, where the disease is classically recognized, bovine coronavirus (BCoV) has been assigned as a major etiologic agent of the disease. Nonetheless, in the Southern hemisphere, an in-deep etiological survey on WD cases had not been carried out. This study aimed to survey for BCoV by nested-RT-PCR, rotavirus by polyacrylamide gel electrophoresis (PAGE) and ELISA, bacteria by classical bacteriological methods and PCR for virulence factors and parasites by sugar flotation test on fecal samples of 21 cows from a farm during an outbreak of WD in São Paulo state, Southeastern Brazil. BCoV was detected in all 21 samples, while rotavirus was detected in two symptomatic cows. Escherichia coli, Yersinia intermedia, Providencia rustigianii Proteus penneri, Klebsiella terrigena and Enterobacter aglomerans were detected in samples from both asymptomatic and healthy cows in different associations. The study of E. coli virulence factors revealed that the strains isolated were all apathogenic. Cysts of Eimeria sp. and eggs of Strongyloidea were detected at low numbers in four of the symptomatic cows, with one co-infestation. These results suggest BCoV as the main etiologic agent of the cases of WD in Brazil, a conclusion that, with the clinical and epidemiological patterns of the disease studied herein, match those already described elsewhere. These findings give basis to the development of preventive measures and contribute to the understanding of the etiology of WD.