Anticorpos anti Brucella abortus, anti Brucella canis e anti Leptospira spp. em raposas (Pseudalopex vetulus) do semiárido paraibano, Nordeste do Brasil

O objetivo do presente trabalho foi determinar a ocorrência de anticorpos anti Brucella abortus, anti Brucella canis e anti Leptospira spp. em raposas (Pseudalopex vetulus). Para tanto, foram utilizadas 60 raposas atropeladas em rodovias no semiárido da Paraíba, Nordeste do Brasil. Para a detecção de anticorpos anti Brucella abortus, o teste do antígeno acidificado tamponado (AAT) foi empregado como teste de triagem, e a prova do 2-mercaptoetanol foi empregada como método confirmatório. Para o diagnóstico sorológico das infecções por Brucella canis e Leptospira spp., foram utilizados os testes de imunodifusão em gel de agar (IDGA) e soroaglutinação microscópica, respectivamente. Todas as amostras foram negativas na pesquisa de anticorpos anti Brucella canis e anti Leptospira spp. Das 60 raposas testadas, 16 (26,6%) foram positivas para anticorpos anti Brucella abortus no teste de AAT, e quatro (6,7%) amostras foram confirmadas no teste de 2-mercaptoetanol, sendo duas amostras com título 100 e duas com título 50.

Pesquisa de aglutininas anti Brucella canis em soros humanos na cidade de São Paulo, Brasil

De 330 soros humanos examinados pela prova de soroaglutinação lenta em tubos, 4(1,21%) apresentaram aglutininas anti Brucella canis em diluição 1:100 (1 reagente com título 100, 2 reagentes com título 200 e 1 reagente com título 400).

Brucella canis: inquéritos sorológico e bacteriológico em população felina

De 134 soros de felinos domésticos examinados pela prova de soroaglutinação lenta em tubos, 4 (3%) foram positivos para Brucella canis, todos com título igual a 100. Não se obteve êxito na tentativa de isolamento de Brucella canis através de hemocultura desses animais.

Inquérito sorológico e fatores de risco para a brucelose por Brucella canis em cães do município de Santana de Parnaíba, Estado de São Paulo

Foi investigada a prevalência da brucelose causada por Brucella canis em cães do município de Santana de Parnaíba, SP, Brasil, e realizado um estudo de possíveis fatores de risco associados à soropositividade para B. canis. Foram examinadas 410 amostras de soro sanguíneo de cães colhidas durante a campanha de vacinação anti-rábica animal, realizada em agosto de 1999. A imunodifusão em gel de ágar (IDGA), utilizando antígeno de lipopolissacarídeos e proteínas de Brucella ovis, amostra Reo 198, foi empregada em soros normais como teste de triagem, e, para a confirmação, a mesma técnica foi aplicada em soros tratados pelo 2-mercaptoetanol (IDGA-ME). A reação de fixação de complemento (CFT), utilizando antígeno de B. ovis, amostra 63/290, também foi utilizada como prova confirmatória. A determinação da prevalência considerou como positivos os animais que reagiram positivamente nos dois testes confirmatórios (IDGA-ME e CFT). A prevalência da B. canis foi de 2,2% (I.C. 95% = 1,01-4,13%). A análise estatística mostrou que os cães com acesso irrestrito à rua o dia todo (manejo do tipo solto) estiveram mais expostos ao risco da infecção por B. canis, com um valor de odds ratio de 8,73 (I.C. 95% = 1,48-51,55) e p=0,04.

Detección molecular y aislamiento de Brucella canis en perros del área metropolitana de Monterrey, N.L.

Tesis (Maestro en Ciencias Veterinarias) U.A.N.L., 2006

Validation of an ELISA method for the serological diagnosis of canine brucellosis due to Brucella canis

In the present study, the validation of an enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of canine brucellosis is described. Two different antigenic extracts, obtained by heat or ultrasonic homogenization of microbial antigens from a wild isolate of Brucella canis bacteria, were compared by ELISA and Western blot (WB). A total of 145 canine sera were used to define sensitivity, specificity and accuracy of the ELISA as follows: (1) sera from 34 animals with natural B. canis infection, confirmed by blood culture and PCR, as well as 51 sera samples from healthy dogs with negative results by the agar-gel immunodiffusion (ACID) test for canine brucellosis, were used as the control panel for B. cants infection; and (2) to scrutinize the possibility of cross reactions with other common dog infections in the same geographical area in Brazil, 60 sera samples from dogs harboring known infections by Leptospira sp., Ehrlichia canis, canine distemper virus (CDV), Neospora caninum, Babesia canis and Leishmania chagasi (10 in each group) were included in the study. The ELISA using heat soluble bacterial extract (HE-antigen) as antigen showed the best values of sensitivity (91.18%), specificity (100%) and accuracy (96.47%). In the WB analyses, the HE-antigen showed no cross-reactivity with sera from dogs with different infections, while the B. canis sonicate had various protein bands identified by those sera. The performance of the ELISA standardized with the heat soluble B. canis antigen indicates that this assay can be used as a reliable and practical method to confirm infection by this microorganism, as well as a tool for seroepidemiological studies. (C) 2010 Elsevier Ltd. All rights reserved.

A polymerase chain reaction for the detection of Brucella canis in semen of naturally infected dogs

The objective was to evaluate a PCR assay for the detection of Brucella canis in canine semen, comparing its performance with that of bacterial isolation, serological tests and PCR assay of blood. Fifty-two male dogs were examined clinically to detect reproductive abnormalities and their serum was tested by the rapid slide agglutination test, with and without 2-mercaptoethanol (2ME-RSAT and RSAT, respectively). In addition, microbiological culture and PCR assays were performed on blood and semen samples. The findings of the semen PCR were compared (Kappa coefficient and McNemar test) to those of blood PCR, culture of blood and semen, RSAT, and 2ME-RSAT. Nucleic acid extracts from semen collected from dogs not infected with B. canis were spiked with decreasing amounts of B. canis RM6/66 DNA and the resulting samples subjected to PCR. In addition, semen samples of non-infected dogs were spiked with decreasing amounts of B. canis CFU and the resulting suspensions were used for DNA extraction and amplification. of the 52 dogs that were examined, the following tests were positive: RSAT, 16 (30.7%); 2ME-RSAT, 5 (9.6%); blood culture, 14 (26.9%); semen culture, 11 (21.1%); blood PCR, 18 (34.6%); semen PCR, 18 (34.6%). The PCR assay detected as few as 3.8 fg of B. canis DNA experimentally diluted in 444.9 ng of canine DNA (extracted from semen samples of noninfected dogs). In addition, the PCR assay amplified B. canis genetic sequences from semen samples containing as little as 1.0 x 10(0) cfu/mL. We concluded that PCR assay of semen was a good candidate as a confirmatory test for the diagnosis of brucellosis in dogs; its diagnostic performance was similar to blood culture or blood PCR. Furthermore, the PCR assay of semen was more sensitive than the 2ME-RSAT or semen culture. Examination of semen by PCR should be included for diagnosis of brucellosis prior to natural mating or AI; in that regard, some dogs that were negative on serological and microbiological examinations as well as blood PCR were positive on PCR of semen. (c) 2007 Elsevier B.V. All rights reserved.

A polymerase chain reaction for detection of Brucella canis in vaginal swabs of naturally infected bitches

A PCR assay for the detection of Brucella canis in canine vaginal swab samples was evaluated, comparing its performance with that of bacterial isolation, serological tests, and a blood PCR assay. One hundred and forty-four female dogs were clinically examined to detect reproductive problems and they were tested by the rapid slide agglutination test, with and without 2-mercaptoethanol (2ME-RSAT and RSAT, respectively). In addition, microbiological culture and PCR were performed on blood and vaginal swab samples. The results of the vaginal swab PCR were compared to those of the other tests using the Kappa coefficient and McNemar test. of the 144 females that were examined, 66 (45.8%) were RSAT positive, 23 (15.9%) were 2ME-RSAT positive, 49 (34.02%) were blood culture positive, 6 (4.1%) were vaginal swab culture positive, 54 (37.5%) were blood PCR positive, 52 (36.2%) were vaginal swab PCR positive, and 50.69% (73/144) were positive by the combined PCR. The PCR was able to detect as few as 3.8 fg of B. canis DNA experimentally diluted in 54 ng of canine DNA, extracted from vaginal swab samples of non-infected bitches. In addition, the PCR assay amplified B. canis genetic sequences from vaginal swab samples containing 1.0 x 10(0) cfu/mL. In conclusion, vaginal swab PCR was a good candidate as a confirmatory test for brucellosis diagnosis in bitches suspected to be infected, especially those negative on blood culture or blood PCR; these animals may be important reservoirs of infection and could complicate attempts to eradicate the disease in confined populations. (C) 2007 Elsevier B.V. All rights reserved.

Perfil sorológico, isolamento bacteriano e valores hematológicos e urinários em cães naturalmente infectados com Brucella canis

Descrevem-se os parâmetros hematológicos, urinários, perfil sorológico de aglutininas antibrucélicas e resultados de isolamento bacteriano de swab vaginal, líquido prostático e hemocultura de 12 cães naturalmente infectados por Brucella canis. Observaram-se flutuação dos resultados sorológicos, ausência de isolamento de B. canis nos diversos materiais colhidos e valores hematológicos e urinários predominantemente normais. Discute-se o diagnóstico de brucelose canina em nível individual.

Ocorrência de anticorpos anti-Brucella abortus e anti-Brucella canis em cães rurais e urbanos do Município de Monte Negro, Rondônia, Brasil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Brazil Anticorpos anti-Brucella canis e anti-Brucella abortus em caes de Araguaina, Tocantins

The aims of the present study were to determine the seroprevalence of infection by Brucella canis and Brucella abortus and to evaluate possible risk factors for infection in dogs from Araguaina, Tocantins, Brazil. Sera from 374 dogs, of the urban zones of the municipality, from both sexes, were submitted to the agar-gel immunodiffusion for Brucella canisantibodies and to rose Bengal test (AAT) and fluorescence polarization assay (FPA) for Brucella abortus-antibodies. From the 374 tested dogs, 21 reacted in the AAT, but no one was positive in the FPA. The seroprevalence of B. canis infection found in Araguaina, Tocantins, Brazil, was 44.53% (95% IC; 39.43 to 49.72). No association was found among seropositivity for B. canis and the risk factors studied. Thus, data from the present study showed that there was no infection by B. abortus among dogs in the sample and that infection by B. canis is widespread and at high prevalence in Araguaina, Tocantins, Brazil.

Risk factors and presence of antibodies to Brucella canis and smooth Brucella in dogs from the municipality of Araguaina, Tocantins, Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Transtornos reprodutivos causados por agentes infecciosos em animais de canis comerciais da microrregião de Ribeirão Preto, estado de São Paulo

Pós-graduação em Medicina Veterinária - FCAV