Identification of enteric viruses circulating in a dog population with low vaccine coverage

Although the use of vaccines has controlled enteric diseases in dogs in many developed countries, vaccine coverage is still under optimal situation in Brazil. There is a large population of nonimmunized dogs and few studies about the identification of the viruses associated with diarrhea. To address this situation, stool samples from 325 dogs were analyzed by polymerase chain reaction for the detection of common enteric viruses such as Canine adenovirus (CAdV), Canine coronavirus (CCoV), Canine distemper virus (CDV), Canine rotavirus (CRV) and Carnivorous protoparvovirus 1 (canine parvovirus 2; CPV-2). At least one of these species was detected in 56.6% (184/325) of the samples. The viruses detected most frequently in either diarrheic or nondiarrheic dog feces were CPV-2 (54.3% of the positive samples), CDV (45.1%) and CCoV (30.4%), followed by CRV (8.2%) and CAdV (4.9%). Only one agent was detected in the majority of the positive samples (63%), but co-infections were present in 37% of the positive samples and mainly included CDV and CPV-2. The data presented herein can improve the clinical knowledge in regions with low vaccine coverage and highlight the need to improve the methods used to control these infectious diseases in domestic dogs.(AU)

A Multiplex real-time PCR for detection of Mycoplasma gallisepticum and Mycoplasma synoviae in clinical samples from Brazilian commercial poultry flocks

Mycoplasma gallisepticum (MS) and Mycoplasma synoviae (MS) are important avian pathogens and cause economic losses to the poultry industry. Molecular biology techniques are currently used for a rapid detection of these pathogens and the adoption of control measures of the diseases. The aim of this study was to develop and validate a technique for simultaneous detection of MG and MS by multiplex real time polymerase chain reaction (PCR). The complete assay (Multiplex MGMS) was designed with primers and probes specific for each pathogen and developed to be carried out in a single tube reaction. Vaccines, MG and MS isolates and DNA from other Mycoplasma species were used for the development and validation of the method. Further, 78 pooled clinical samples from different poultry flocks in Brazil were obtained and used to determine the sensitivity and specificity of the technique in comparison to 2 real time PCR assays specific for MG (MG PCR) and MS (MS PCR). The results demonstrated an agreement of 100% (23 positive and 44 negative samples) between Multiplex MGMS and MG PCR in the analysis of 67 samples from MG positive and negative poultry flocks, and an agreement of 96.9% between Multiplex MGMS and MS PCR in the analysis of 64 samples from MS positive and negative poultry flocks. Considering the single amplification tests as the gold standard, the Multiplex MGMS showed 100% of specificity and sensitivity in the MG analysis and 94.7% sensitivity and 100% specificity in the MS analysis. This new assay could be used for rapid analysis of MG and MS in the poultry industry laboratories.(AU)

Caracterização antigênica e fenotípica de cepas de Pasteurella multocida isoladas de pulmões de suínos com pneumonia e/ou pleurite / Antigenic and phenotypic characterization of Pasteurella multocida strains isolated from the lungs of pigs with pneumonia and/or pleuritic lesions

Foi analisada a variabilidade antigênica e fenotípica de 22 cepas de Pasteurella multocida isoladas de pulmões de suínos com pneumonia e/ou pleurite. Os testes fenotípicos foram realizados pela determinação de características bioquímicas e sensibilidade a agentes antimicrobianos. Todos os isolados fermentaram manitol e sorbitol, mas nenhum arabinose; 14 foram capazes de metabolizar xilose, quatro trealose, dois dulcitol e um maltose. A análise destas características permitiu agrupar os isolados em 5 padrões bioquímicos distintos. Quanto à sensibilidade a nove agentes antimicrobianos, verificou-se grande variação, com apenas 50 por cento dos isolados sensíveis a pelo menos sete dos nove antibióticos testados. Nenhum princípio ativo foi capaz de inibir todos os isolados. A melhor eficiência foi observada com a amoxicilina (30 mg); 72,7 por cento dos isolados se mostraram sensíveis. A menor eficiência foi demonstrada pela espectinomicina (100 mg) com 45,5 por cento. A caracterização antigênica consistiu na sorotipagem capsular e determinação de variabilidade do gene de proteína de membrana externa (ompH) pela reação em cadeia da polimerase (PCR) e digestão com cinco enzimas de restrição. Das 22 cepas, 21 foram compatíveis com sorotipo capsular A e uma com D. A caracterização do gene ompH agrupou os isolados em sete padrões distintos que apresentaram boa correlação com os testes bioquímicos (AU)

Isolamento de Arcobacter butzleri de músculos de carcaças de suínos de terminação e de matrizes descartadas abatidos em um matadouro no Estado do Rio Grande do Sul, Brasil

Portions of muscle from carcasses of 74 fattening pigs and 150 culled sows were collected from abattoir in the State of Rio Grande do Sul, Brazil. The samples were inoculated into EMJH (Ellinghausen, MacCullough, Johnson & Harris) medium and the culture filtered through 0.45µm membrane on blood agar. Arcobacter sp were isolated, 69 isolates, typed by PCR as Arcobater butzleri, respectively from fattening pigs (21 isolates, 29.3%) and culled sows (48 isolates, 32%).

Foram colhidos fragmentos de músculos da carcaça de 74 suínos de abate e de 150 porcas descartadas de granjas de ciclo completo, abatidos em frigorífico no Rio Grande do Sul. Os materiais foram inoculados em meio EMJH (Ellinghausen, MacCullough, Johnson & Harris), incubados a 30ºC e o cultivo foi passado a placas de agar sangue através de membrana de 0,45µm. Foram obtidas 69 amostras de Arcobacter sp, classificadas por PCR como A. butzleri, respectivamente de suínos de abate (21 isolamentos, 29,3%) e matrizes descartadas (48 isolamentos, 32%).

Isolation of Arcobacter spp from poultry carcasses, in Brazil

Fourty eight isolates of Arcobacter spp were obtained from 37 poultry carcasses, from abattoir, among 80 carcasses examined. Attempts for culturing were made from the skin and muscle, resulting on 25 positive cultures from muscle and 23 from skin. Classification was achieved by phenotypic characterization and PCR and multiplex PCR, resulting 41 samples of Arcobacter butzleri and 07 Arcobacter sp. This is the first report on the occurrence of Arcobacter in animal carcasses in Brazil.

Foram isoladas 48 amostras de Arcobacter spp de 37 carcaças de frangos colhidas em frigorífico, prontas para consumo, entre 80 carcaças examinadas. Foram feitas tentativas de cultivo a partir de pele e de músculo, sendo obtidas 25 cultivos positivos de músculo e 23 de pele. As bactérias foram classificadas pelas características fenotípicas e pelo teste de PCR e PCR múltiplo, obtendo-se 41 amostras classificadas como Arcobacter butzleri e 07 com classificação a nível de gênero Arcobacter sp. Estes são os primeiros relatos sobre a ocorrência das bactérias em carcaças de animais no Brasil.

Isolation of Arcobacter spp from poultry carcasses, in Brazil

Fourty eight isolates of Arcobacter spp were obtained from 37 poultry carcasses, from abattoir, among 80 carcasses examined. Attempts for culturing were made from the skin and muscle, resulting on 25 positive cultures from muscle and 23 from skin. Classification was achieved by phenotypic characterization and PCR and multiplex PCR, resulting 41 samples of Arcobacter butzleri and 07 Arcobacter sp. This is the first report on the occurrence of Arcobacter in animal carcasses in Brazil.

Foram isoladas 48 amostras de Arcobacter spp de 37 carcaças de frangos colhidas em frigorífico, prontas para consumo, entre 80 carcaças examinadas. Foram feitas tentativas de cultivo a partir de pele e de músculo, sendo obtidas 25 cultivos positivos de músculo e 23 de pele. As bactérias foram classificadas pelas características fenotípicas e pelo teste de PCR e PCR múltiplo, obtendo-se 41 amostras classificadas como Arcobacter butzleri e 07 com classificação a nível de gênero Arcobacter sp. Estes são os primeiros relatos sobre a ocorrência das bactérias em carcaças de animais no Brasil.

Isolamento de Arcobacter butzleri de músculos de carcaças de suínos de terminação e de matrizes descartadas abatidos em um matadouro no Estado do Rio Grande do Sul, Brasil

Portions of muscle from carcasses of 74 fattening pigs and 150 culled sows were collected from abattoir in the State of Rio Grande do Sul, Brazil. The samples were inoculated into EMJH (Ellinghausen, MacCullough, Johnson & Harris) medium and the culture filtered through 0.45µm membrane on blood agar. Arcobacter sp were isolated, 69 isolates, typed by PCR as Arcobater butzleri, respectively from fattening pigs (21 isolates, 29.3%) and culled sows (48 isolates, 32%).

Foram colhidos fragmentos de músculos da carcaça de 74 suínos de abate e de 150 porcas descartadas de granjas de ciclo completo, abatidos em frigorífico no Rio Grande do Sul. Os materiais foram inoculados em meio EMJH (Ellinghausen, MacCullough, Johnson & Harris), incubados a 30ºC e o cultivo foi passado a placas de agar sangue através de membrana de 0,45µm. Foram obtidas 69 amostras de Arcobacter sp, classificadas por PCR como A. butzleri, respectivamente de suínos de abate (21 isolamentos, 29,3%) e matrizes descartadas (48 isolamentos, 32%).