Abstract The aim of this study was to detect the IS900 region of Mycobacterium avium subsp. paratuberculosis (MAP) in bovine milk samples using real-time polymerase chain reaction (qPCR) and conventional PCR, and to study the agreement between these tests. A total of 121 bovine milk samples were collected from herds considered positive for MAP, from the State of Pernambuco, Brazil. MAP DNA was detected in 20 samples (16.5%) using conventional PCR and in 34 samples (28.1%) using qPCR. MAP DNA was detected in all of the 6 animal farms studied. Moderate agreement was found between qPCR and conventional PCR results, where the sensitivity and specificity of conventional PCR in relation to qPCR were 50% and 96.6%, respectively. Thus, the IS900 region of MAP was found in bovine milk samples from the State of Pernambuco. To the best of our knowledge, this is the first report of MAP DNA found in bovine milk in Northeast Brazil. We also demonstrated the qPCR technique is more sensitive than conventional PCR with respect to detection of MAP in milk samples.

Resumo A identificação de Listeria spp. em áreas de manipulação de alimentos é de grande preocupação para as agências de vigilância da saúde. O seu controle, muitas vezes, é prejudicado pela capacidade da bactéria em crescer e se manter no ambiente mesmo sob condições adversas. Objetivou-se, com este estudo, isolar e identificar Listeria spp. em áreas de manipulação de estabelecimentos varejistas de alimentos no Estado de Pernambuco, Brasil. Foram colhidas 86 amostras de swabs, procedentes de equipamentos, utensílios e instalações utilizados para processamento de embutidos cárneos, distribuídas em dez estabelecimentos varejistas. Para identificação de Listeria spp., utilizaram-se os métodos rápidos 3M™, Quick Swab e 3M™ Petrifilm™. A confirmação de Listeria monocytogenes foi realizada pela técnica de Reação em Cadeia da Polimerase em Tempo Real (qPCR). Nos estabelecimentos, foi realizada uma lista de verificação para avaliar as condições higiênico-sanitárias dos pontos de coletas. Das 86 amostras analisadas, 27 (31,2%) [intervalo de confiança 21,81% a 42,30%] foram positivas para Listeria spp. e, destas, uma (3,7%) para Listeria monocytogenes. Os pontos de maior contaminação foram o piso (50,0%), a placa de polietileno (42,9%) e a faca (40,0%). Dos dez estabelecimentos analisados, observou-se que oito (80,0%) tiveram ao menos uma amostra positiva para Listeria spp. Ao analisar a associação entre as condições higiênico-sanitárias dos pontos de coletas e o exame microbiológico, foi constatado não haver diferença significativa (p = 0,700). Diante dos resultados obtidos, conclui-se que a Listeria spp. está amplamente distribuída nos ambientes estudados, o que pode favorecer a contaminação dos alimentos e, consequentemente, oferecer riscos à saúde pública. Desta forma, recomenda-se que parâmetros microbiológicos devem ser estabelecidos pelos órgãos reguladores de saúde para alimentos cárneos e seus embutidos, com o intuito de oferecer um alimento seguro para os consumidores.

Abstract The identification of Listeria spp. in food handling areas is of great concern to health surveillance agencies, and their control is often hampered by the ability of the bacteria to grow and maintain themselves even under adverse conditions. The present study aimed to isolate and identify Listeria spp. in the food handling areas of 10 retail food stores in the state of Pernambuco, Brazil. Eighty-six swab samples were collected from equipment, utensils and surfaces used for processing ready-to-eat meat products. The Dry and Wet Swabbing Methods (3M™ Quick Swabs) and 3M™ Petrifilm™ Plates were used to identify Listeria spp. Contamination by Listeria monocytogenes was confirmed by the Real-time Polymerase Chain Reaction (qPCR). The hygienic and sanitary conditions of the food handling areas of each store were also assessed. Listeria spp. was isolated in eight stores (80%). Of the 86 swab samples analyzed, 27 (31.2%) [confidence interval 21.81% to 42.30%] were positive for Listeria spp. and only one (3.7%) was confirmed as Listeria monocytogenes. The main contamination sites were the floor (50.0%), the plastic cutting board (42.9%) and the knife (40.0%). None of the hygienic and sanitary conditions assessed in the present study were associated with contamination by Listeria spp. (p = 0.700). It was concluded that Listeria spp. was widely distributed in the retail food stores studied, being a possible risk factor for public health.

Resumo O objetivo deste estudo foi pesquisar a ocorrência de Toxoplasma gondii em ovinos abatidos no Estado de Alagoas, Brasil por meio de diferentes técnicas de diagnóstico. Foram utilizadas amostras de soros e tecidos de 100 ovinos abatidos. Para a pesquisa de anticorpos foi utilizada a Reação de Imunofluorescência Indireta (RIFI), e os tecidos dos animais soropositivos (ponto de corte ≥1:64) foram submetidos às técnicas de Reação de Cadeia da Polimerase (PCR) e Imunohistoquímica (IHQ). Para o estudo da concordância entre as técnicas diretas foi empregado o teste Kappa. Na RIFI, 14% (14/100) das amostras foram soro-positivas. Na PCR, 21,43% (3/14) dos animais foram positivos e, na IHC, 7,14% (1/14) apresentaram marcação positiva para T. gondii no tecido cerebral. Na histopatologia, o achado predominante foi o infiltrado celular mononuclear no coração e manguito perivascular no cérebro e cerebelo. A concordância entre as técnicas diretas de diagnóstico foi moderada (K= 0,44). Desse modo, é importante utilizar diferentes técnicas diretas no diagnóstico da toxoplasmose em ovinos naturalmente infectados.

Abstract The aim of this study was to investigate occurrence of Toxoplasma gondii in sheep slaughtered in the state of Alagoas, Brazil, by means of different diagnosis techniques. Serum samples and tissues from 100 slaughtered sheep were used. To detect antibodies, the indirect immunofluorescence antibody test (IFAT) was used, and tissues from seropositive animals (cut-off ≥1:64) were submitted to Polymerase Chain Reaction (PCR) and immunohistochemistry (IHC). To assess the concordance between the direct techniques, the kappa test was used. In the IFAT, it was observed that 14% (14/100) of the ovine samples were serum-positive. In the PCR, 21.43% (3/14) of the animals were positive and in IHC, it was observed that 7.14% (1/14) were positively stained for T. gondii in cerebral tissue. Histopathologically, the predominant finding was the presence of mononuclear cell infiltrate in the heart and a perivascular cuff in the cerebrum and cerebellum. The concordance between the direct diagnosis techniques was moderate (k=0.44). Thus, it is important to use different direct techniques in diagnosing toxoplasmosis in naturally infected sheep.

The present study reports the first isolation of Actinobacillus seminis from a goat in Brazil. A four-year-old Moxotó breeding goat in a flock of 70 goats and 65 sheep reared together in the county of Patos, semiarid region of Northeastern Brazil, showed clinical signs of unilateral orchitis and epididymitis. Diagnosis of A. seminis infection was confirmed by association of clinical findings, bacterial isolation and 16S rRNA gene sequencing. This result suggests that A. seminis may be an additional cause of infertility in goats, and that sheep may be the source of infection because the mixed farming system allows the contact between sheep and goats in the semiarid region of Northeastern Brazil.

A study was conducted to verify the presence of mycoplasmas and ureaplasmas DNA in sheep semen samples from the State of Pernambuco. The PCR assay was conducted of according with standard protocols with generic primers. Mollicutes DNA was detected in 26.0% and Ureaplasma spp. in 12.0% of semen samples.

O objetivo do presente trabalho foi determinar a ocorrência de anticorpos anti-Brucella rugosa e anti-Brucella lisa em cães do município de Natal, Estado do Rio Grande do Norte, Brasil, bem como identificar fatores de risco associados à positividade e realizar a detecção molecular em animais soropositivos. Foram utilizados soros sanguíneos de 416 cães atendidos em clínicas veterinárias durante o período de março a novembro de 2011. Para o diagnóstico sorológico da infecção por Brucella rugosa, foi empregada a prova de imunodifusão em gel de ágar (IDGA), utilizando antígeno de lipopolissacarídeos e proteínas de Brucella ovis, amostra Reo 198 e, para o diagnóstico da infecção por Brucella lisa, foi utilizado o teste do antígeno acidificado tamponado (AAT). De animais soropositivos, foram coletadas amostras de sangue com citrato de sódio para o diagnóstico pela reação em cadeia pela polimerase (PCR). A frequência de anticorpos anti-Brucella rugosa foi de 28,9% (120/416). Todos os animais foram negativos para anticorpos anti-Brucella lisa. Dentre 80 animais soropositivos, o DNA de Brucella spp. foi amplificado em três animais (3,8%). Não foram identificados fatores de risco associados à soropositividade. Conclui-se que a infecção por Brucella rugosa está presente no município de Natal, bem como se sugere o monitoramento sorológico de animais atendidos em clínicas visando à identificação de fontes de infecção.

The aim of this study was to determine the occurrence of anti-rough Brucella and anti-smooth Brucella antibodies in dogs from the county of Natal, Rio Grande do Norte state, Brazil, as well as to identify risk factors associated with positivity and to perform molecular detection of the agent in seropositive animals. Sera from 416 dogs attended in veterinary clinics during the period from March to November 2011 were used. For the serological diagnosis of rough Brucella the agar gel immunodiffusion (AGID) test, using antigen of lipopolysaccharides and proteins from Brucella ovis, strain Reo 198, was carried, and for smooth Brucella the buffered plate agglutination test (BPAT) was used. From seropositive animals, blood samples with sodium citrate were collected for the diagnosis by polymerase chain reaction (PCR). Frequency of anti-rough Brucella antibodies was 28.9% (120/416). All animals were negative for anti-smooth Brucella antibodies. Of the 80 seropositive animals Brucella spp. DNA was amplified in three (3.8%). Risk factors associated with the seropositivity were not identified. It was concluded that rough Brucella infection is present in the county of Natal, as well as it is suggested the serological monitoring of animals attended at clinics aiming the identification of sources of infection.