Survey for Leptospira and Brucella in Amazonian manatees, Amazon river dolphins, and a tucuxi in the Brazilian Amazon.

Leptospirosis and brucellosis are zoonotic diseases with global distributions that represent severe hazards to humans and animals. We investigated exposure to Leptospira spp. and Brucella spp. in samples from Amazonian manatees Trichechus inunguis, Amazon river dolphins Inia geoffrensis, and a tucuxi Sotalia fluviatilis. The animals were free-ranging or undergoing in situ rehabilitation in the mid-Solimões River region, Brazilian Amazon. Serum samples from 19 Amazonian manatees were tested by microscopic agglutination test, Rose Bengal test, and 2-mercaptoethanol Brucella agglutination test. Antibodies against Leptospira spp. were detected in 63% of the manatees tested and serovar Patoc was considered the infecting serovar in all positive samples. Titers were generally low, indicating chronic exposure, but higher titers indicative of an active infection were detected in 3 animals. Anti-Brucella spp. antibodies were not detected. Tissue and/or body fluid samples from 12 Amazon river dolphins, a tucuxi, and 2 Amazonian manatees were investigated by multiplex PCR and bacteriology for Leptospira spp. and Brucella spp. All samples were negative. However, Enterococcus faecalis was isolated from uterine fluid, lymph node, and lung of 3 Amazon river dolphins. Bacillus spp. were isolated from milk and synovial fluid from 2 Amazon river dolphins and from a milk sample from 1 Amazonian manatee. Knowledge of the pathogens present in Amazonian manatees, Amazon river dolphins, and tucuxis is of great relevance to species conservation and environmental health. Although no clinical signs were noted, further research is needed to elucidate the clinical relevance of infection by Leptospira sp. serovar Patoc in Amazonian aquatic mammals.

Causes of equine abortion, stillbirth, and perinatal mortality in Brazil / Causas de abortamento, natimortalidade e mortalidade em equinos no Brasil

Abortion and complications in reproduction are important causes of economic loss in horse breeding. Studies of its causal agents can help to identify the primary pathogens or other factors involved and define appropriate measures to reduce its occurrence. This research aimed to investigate the primary causes of equine abortion, stillbirth, and perinatal mortality in regions of Brazil. Tissue from aborted fetuses, stillbirths, neonates and foals submitted to the Biological Institute of São Paulo, Brazil, from January 2010 to July 2013 were processed for viral and bacterial isolation, polymerase chain reaction (PCR), histology, and immunohistochemistry. Bacterial infection was the primary detected cause of abortion, found in 16 of the 53 animals submitted for bacterial analysis followed by viruses analysis in 2 of 105 animals, and noninfectious causes (neonatal isoerythrolysis) in 2 of 105 animals. Fungi were found in a single sample of 53 tested. The most frequent bacteria recovered were Escherichia coli, Enterobacter aerogenes, combined E. coli and Streptococcus spp., Staphylococcus spp., and Bacillus spp. The following agents were each observed in a single sample: Arcanobacterium pyogenes, Streptococcus spp., Corynebacterium spp., Actinobacillus spp., and Rhodococcus equi. The predominant identification of fecal and other opportunistic bacteria as opposed to pathogens commonly associated with equine abortion, such as Leptospira spp. and equine herpesvirus type 1 (EHV-1), suggests the need of improving hygiene management of breeding mares to prevent bacterial infection that may cause fetal loss, stillbirth, and perinatal mortality.(AU)

Abortamento e complicações na reprodução são importantes causas de perda econômica na equideocultura. Estudos dos agentes causais podem ajudar a identificar patógenos ou outros fatores envolvidos e definir medidas apropriadas para reduzir sua ocorrência. Esta pesquisa investigou as causas primárias de aborto, natimortalidade e mortalidade perinatal em equinos de diversas regiões do Brasil. Tecidos de fetos abortados, natimortos e potros submetidos ao Instituto Biológico de São Paulo, Brasil, no período de janeiro de 2010 a julho de 2013, foram processados por meio de técnicas de isolamento viral e bacteriano, PCR, histologia e imuno-histoquímica. Infecção bacteriana foi a causa mais detectada, encontrada em 16 de 53 amostras submetidas à análise bacteriana, seguida de causa viral em 2 de 105 amostras, e causas não infecciosas (isoeritrólise neonatal) em 2 de 105 amostras. Fungo foi encontrado em uma única amostra de 53 testadas. As bactérias isoladas mais frequentemente foram Escherichia coli, Enterobacter aerogenes, E. coli associada a Streptococcus spp., Staphylococcus spp. associado a Bacillus spp. Os seguintes agentes foram observados em uma única amostra cada: Arcanobacterium pyogenes, Streptococcus spp., Corynebacterium spp., Actinobacillus spp. e Rhodococcus equi. A identificação predominante de bactérias fecais e outras bactérias oportunistas, ao invés de outros patógenos comumente associados a quadros de abortamento equino, tais como Leptospira spp. e Herpesvírus equino tipo 1, sugere a necessidade de maior atenção no manejo higiênico das éguas em reprodução, a fim de prevenir infecções bacterianas que possam causar perda fetal, natimortalidade e mortalidade perinatal.(AU)

Detection of CDT toxin genes in Campylobacter spp. strains isolated from broiler carcasses and vegetables in São Paulo, Brazil

Campylobacteriosis is a worldwide distributed zoonosis. One of the main virulence factors related to Campylobacter spp. in animals and humans is the cytolethal distending toxin (CDT), encoded by three adjacent genes (cdtA, cdtB, cdtC). The occurrence of Campylobacter spp. in samples of vegetables has not been reported in Brazil yet, and has seldom been described in the international literature. The detection of CDT in these strains has not been reported, either. The objectives of the present study were to determine the occurrence of Campylobacter spp. strains carrying virulence factors in samples of poultry and vegetables (lettuce and spinach) from different points of sale, thus verifying if vegetables are as an important vehicle for potentially virulent Campylobacter spp. strains as poultry. Twenty four strains were identified as Campylobacter jejuni by phenotypic and genotypic methods: 22 from broiler carcasses and two from lettuce samples. Three strains were identified as Campylobacter coli: two from broiler carcasses and one from lettuce. The presence of the cdt genes were detected in 20/24 (83.3%) C. jejuni strains, and 3/3 (100%) C. coli strains. The isolation of Campylobacter spp. strains with the cdt gene cluster in lettuce samples points to a new possible source of contamination, which could have an impact in the vegetable production chain and risk to public health. Results show that potentially virulent C. jejuni and C. coli strains remain viable in samples of broiler carcasses and vegetables at the points of sale.

Caracterização epidemiológica e fatores de risco associados à infecção por Chlamydophila abortus em ovinos deslanados do semiárido brasileiro / Epidemiological characterization and risk factors associated with Chlamydophila abortus infection in sheep in Brazilian semiarid

O objetivo do presente trabalho foi determinar as prevalências de propriedades positivas e animais soropositivos para Chlamydophila abortus em ovinos deslanados da região semiárida do Nordeste do Brasil, bem como identificar fatores de risco. Foram colhidas amostras de sangue de 476 ovinos procedentes de 72 propriedades em 14 municípios na mesorregião do Sertão, Estado da Paraíba. Para o diagnóstico sorológico da infecção por Chlamydophila abortus foi utilizada a reação de fixação de complemento (RFC). Uma propriedade foi considerada positiva quando apresentou pelo menos um animal soropositivo. Das 72 propriedades usadas 38 (52,8%) apresentaram pelo menos um animal soropositivo, e dos 476 animais 94 (19,7%) foram soropositivos. Participação em exposições (odds ratio =4,31; IC 95% =1,80-10,35; p=0,011) foi identificada como fator de risco. Sugere-se que a infecção por Chlamydophila abortus encontra-se disseminada em ovinos da região, e baseando-se na análise de fatores de risco, recomenda-se o controle sanitário nas exposições de animais.

The aim of this investigation was to determine the flock-level and animal-level prevalences of Chlamydophila abortus in sheep from the semiarid region of Northeastern Brazil, as well as to identify risk factors. Blood samples were collected from 476 sheep of 72 flocks in 14 counties in the Sertão mesoregion, state of Paraíba. For the serological diagnosis of Chlamydophila abortus infection the complement fixation test (FC) was carried out. A flock was positive when presented at least one seropositive animal. From the 72 flocks, 38 (52.8%) presented at least one seropositive sheep, and 94 (19.7%) of the 476 animals were seropositive. Participation in animal expositions (odds ratio= 4.31; 95% CI= 1.80-10.35; p=0.011) was identified as risk factor. It is suggested that C. abortus infection is spread in sheep of the region, and based on the risk factor analysis sanitary control in animal expositions is recommended.

Detection of CDT toxin genes in Campylobacter spp. strains isolated from broiler carcasses and vegetables in São Paulo, Brazil.

Campylobacteriosis is a worldwide distributed zoonosis. One of the main virulence factors related to Campylobacter spp. in animals and humans is the cytolethal distending toxin (CDT), encoded by three adjacent genes (cdtA, cdtB, cdtC). The occurrence of Campylobacter spp. in samples of vegetables has not been reported in Brazil yet, and has seldom been described in the international literature. The detection of CDT in these strains has not been reported, either. The objectives of the present study were to determine the occurrence of Campylobacter spp. strains carrying virulence factors in samples of poultry and vegetables (lettuce and spinach) from different points of sale, thus verifying if vegetables are as an important vehicle for potentially virulent Campylobacter spp. strains as poultry. Twenty four strains were identified as Campylobacter jejuni by phenotypic and genotypic methods: 22 from broiler carcasses and two from lettuce samples. Three strains were identified as Campylobacter coli: two from broiler carcasses and one from lettuce. The presence of the cdt genes were detected in 20/24 (83.3%) C. jejuni strains, and 3/3 (100%) C. coli strains. The isolation of Campylobacter spp. strains with the cdt gene cluster in lettuce samples points to a new possible source of contamination, which could have an impact in the vegetable production chain and risk to public health. Results show that potentially virulent C. jejuni and C. coli strains remain viable in samples of broiler carcasses and vegetables at the points of sale.

Inquérito sorológico de lentiviroses de pequenos ruminantes (Maedi-Visna e artrite-encefalite caprina) no estado de São Paulo / Serologic survey of small ruminants lentiviruses (Maedi-Visna and caprine arthritis-encephalitis) in São Paulo state

O objetivo deste estudo foi determinar a frequência de animais soropositivos ao vírus da Maedi-Visna (MVV) em ovinos e ao vírus da CAE (CAEV) em caprinos criados no estado de São Paulo. Na pesquisa dos anticorpos séricos anti- MVV e anti-CAEV foi utilizada a técnica de imunodifusão em gel de ágar (IDGA). Dentre os ovinos estudados, 0,3% (4/1235) eram sororreagentes ao MVV e 15,1% caprinos (30/199) ao CAEV. Foi realizada a análise de fatores de risco associados à condição de propriedade positiva para CAEV e Maedi-Visna. Foram selecionadas variáveis para as duas enfermidades, no entanto, quando essas variáveis foram usadas na regressão logística múltipla, não foram identificados fatores de risco para as infecções. A infecção pelo CAEV no estado de São Paulo tem uma ampla disseminação e com uma alta prevalência enquanto que o MVV apresenta baixa prevalência. Ressalta-se a importância de medidas de prevenção e controle para diminuir a ocorrência da CAE e evitar a disseminação da Maedi-Visna.

The aim of this study was to carry out the serological occurrence of Maedi-Visna virus (MVV) and CAE virus (CAEV) in ovines and caprines breeding in São Paulo state. The test to detect MVV and CAEV antibodies was agar gel immunodiffusion (AGID). The detection of antibodies against MVV was 0,3% (4/1235) and against CAEV was 15,1% (30/199). Was carried analysis of risk factors associated with the presence of positive property for CAEV and Maedi-Visna. Variables were selected for both diseases, however, when these variables were used in multivariate logistic regression model were not identified risk factors for the infections. The CAEV infection in the São Paulo state has a wide spread and a high prevalence while MVV has low prevalence. It emphasizes the importance of prevention and control measures to reduce CAEV occurrence and prevent the spread of Maedi-Visna.

Sinais clínicos e ocorrência de anticorpos anti-Chlamydophila abortus em ovinos de São Paulo e Minas Gerais / Clinical signs and occurrence of antibodies anti-Chlamydophila abortus in ovines of São Paulo and Minas Gerais

A Chlamydophila abortusi, anteriormente conhecida como Chlamydia psittaci sovovar 1, é uma bactéria Gram negativa, intracelular obrigatória. Esse micro-organismo é frequentemente encontrado em distúrbios reprodutivos em ovinos, bovinos e caprinos, sendo o aborto epizoótico dos bovinos e o aborto enzoótico dos ovinos e caprinos as manifestações mais importantes. Considerando-se o pouco material literário a respeito da clamidofilose no Brasil, a pesquisa teve como objetivo determinar a presença de anticorpos fixadores de complemento anti-Chlamydophila abortusi, correlacionando os resultados obtidos com achados no exame clínico e histórico dos animais, além de alterações nos índices zootécnicos, em especial na esfera reprodutiva, tais como alto índice de repetição de cio, número elevado de abortamentos, elevado número de natimortos, entre outros. Foram testadas para prova de fixação do complemento 220 amostras de soro de ovinos, de 26 propriedades, distribuídas em 19 municípios, com relato de manifestação reprodutiva, obtendo-se 19,55% (43/220) de testes positivos para Chlamydophila abortusi, com ocorrência de foco constatada de 61,53%. No geral, a titulação de anticorpos encontrada foi baixa, com título não superior a 64. A frequência de manifestação reprodutiva mais observada foi o aborto, representando 65,12% (28/43) do número total de animais soropositivos, seguido de repetição de cio juntamente com nascimento de cordeiro fraco, com frequência de 6,98% (3/43) e, por fim, morte neonatal com 4,65% (2/43), sendo que não houve associação significativa entre animais que foram positivos ao teste e a esses fatores.

The Chlamydophila abortusi was previously known as Chlamydia psittaci sorovar 1, it is a Gram negative and obligate intracellular bacteria. This microorganism is frequently related with reproductive manifestation in ovines, goats and bovines. The major manifestation are Enzootic abortion in bovines and enzootic abortion in small ruminants. There are few literary material about clamidofilosis in Brazil, so the present research had the objective to determine the presence of complement fixing antibodies anti-Chlamydophila abortusi, correlating the results with the clinical examination and historical of the animals, besides variation in the zootecnic index, especially in the reproductive characters like high index of cio repetition, high abortion number, high neonatal mortality number etc. It was tested for the complement fixation test 220 serum ovine samples, from 26 properties, distributed in 19 cities, with historical of reproductive manifestation. It was found 19.55% (43/220) of positive tests to Chlamydophila abortusi, obtaining 61.53% of focus occurrence. In the study low antibodies titers were found, not superior of 64. Abortion was the major reproductive manifestation observed, expressing 65.12% (28/43) of the total number animals serum positive, followed by estrus repetition and weak lamb born, with a frequency of 6.98% (3/43) and, lastly, neonatal mortality death with 4.65% (2/43). No significant association was observed between serum positivity and these factors.

Seroprevalence and risk factors associated with Chlamydophila abortus infection in dairy goats in the Northeast of Brazil / Soroprevalência e fatores de risco associados com a infecção por Chlamydophila abortus em caprinos leiteiros do Nordeste brasileiro

Few data are available on the prevalence and risk factors of Chlamydophila abortus infection in goats in Brazil. A cross-sectional study was carried out to determine the flock-level prevalence of C. abortus infection in goats from the semiarid region of the Paraíba State, Northeast region of Brazil, as well as to identify risk factors associated with the infection. Flocks were randomly selected and a pre-established number of female goats > 12 mo old were sampled in each of these flocks. A total of 975 serum samples from 110 flocks were collected, and structured questionnaire focusing on risk factors for C. abortus infection was given to each farmer at the time of blood collection. For the serological diagnosis the complement fixation test (CFT) using C. abortus S26/3 strain as antigen was performed. The flock-level factors for C. abortus prevalence were tested using multivariate logistic regression model. Fifty-five flocks out of 110 presented at least one seropositive animal with an overall prevalence of 50.0% (95%; CI: 40.3%, 59.7%). Ninety-one out of 975 dairy goats examined were seropositive with titers >32, resulting in a frequency of 9.3%. Lend buck for breeding (odds ratio = 2.35; 95% CI: 1.04-5.33) and history of abortions (odds ratio = 3.06; 95% CI: 1.37-6.80) were associated with increased flock prevalence.

São escassos os trabalhos publicados sobre a prevalência e fatores de risco associados à infecção por Chlamydophila abortus em caprinos no Brasil. Foi conduzido um estudo transversal para determinar a prevalência de rebanhos positivos para a infecção por C. abortus em caprinos do semiárido do Estado da Paraíba, Nordeste do Brasil, bem como identificar os fatores de risco associados com a infecção. Os rebanhos foram selecionados aleatoriamente e um número pré-estabelecido de cabras com idade >12 meses foi amostrado por rebanho. No total, foi colhido sangue de 975 animais procedentes de 110 rebanhos, e no momento da colheita foi aplicado um questionário epidemiológico a cada proprietário. Para o diagnóstico sorológico foi utilizado o teste de fixação de complemento (FC) usando a estirpe de C. abortus S26/3 como antígeno. Os fatores de risco para a prevalência de C. abortus em nível de rebanho foram testados com o uso de modelo de regressão logística multivariada. Cinquenta e cinco rebanhos dos 110 analisados apresentaram pelo menos um animal soropositivo, com uma prevalência de 50,0% (IC 95%: 40,3-59,7%). Noventa e um animais entre os 975 examinados foram soropositivos com título >32, resultando em uma frequência de 9,3%. Compartilhar reprodutores (odds ratio = 2,35; IC 95%: 1,04-5,33) e histórico de abortamentos (odds ratio = 3,06; IC 95%: 1,37-6,80) foram associados com o aumento da prevalência de rebanhos.

Comparative study on PCR and microbiological culture performance for identifying Mycobacterium tuberculosis complex and Mycobacterium bovis specie in bovine samples

The present study aimed at evaluating the concordance between PCR and microbiological culture techniques for analysing organs samples from cattle with suspected lesions of tuberculosis. Fifty-twosamples collected from slaughter houses were analyzed by microbiological culture, and the extracted DNA was amplified by PCR using NZ1 and NZ2 primers. These primers identify the mycobacteria belongingto M. tuberculosis complex, and the primers pair pncA differentiate the M . bovis from M. tuberculosis species. The colonies isolated from 30 samples were suspended, and the extracted DNA was amplifiedby PCR using the same primer pairs. Although the agreement has been considered weak (k = 0.175) between microbiological culture and PCR performed directly in clinical samples using NZ1 and NZ2 primers, the two pairs of primers could amplify the target genes when 100% of the extracted DNA from 30 isolated colonies were used. Thus, PCR employing pncA primer pair enabled to identify M.bovis in the isolated colonies at a short time when compared with the biochemical assays. The concomitant use of PCR and bacteriologic culture techniques hastens the confirmation of detected agent, which is essential inconducting the epidemiological studies and in taking preventive control measures.

Brucella canis infection in dogs attended in veterinary clinics from patos, Paraíba state, Brazil

To determine the frequency of anti-Brucella canis antibodies in dogs attended in veterinary clinics from Patos, Paraíba State, Brazil, as well as to identify risk factors and to isolate and identify the agent, 193 dogs were used. Agar gel immunodiffusion test (AGID) was used to detect B. canis antibodies in sera. Isolation of B. canis was carried out in blood and bone marrow from seropositive animals. Six animals tested seropositive in AGID, resulting in a frequency of 3.11 percent. B. canis was isolated from bone marrow of one seropositive animal, with confirmation by PCR. Lack of cleaning of the dog's environment was identified as risk factor (odds ratio = 7.91). This is the first report of isolation of B. canis in dogs from the Northeast region of Brazil.

Prevalência e fatores de risco associados à infecção por Chlamydophila abortus em granjas suinícolas tecnificadas no estado de Alagoas, Brasil / Prevalence and risk factors associated with infection by Chlamydophila abortus in commercial pig farms in the state of Alagoas, Brazil

Objetivou-se com este estudo calcular a prevalência e identificar os fatores de risco associados à infecção por Chlamydophila abortus em suínos criados em granjas tecnificadas no Estado de Alagoas, Brasil. Para compor a amostra do estudo foram utilizados 342 suínos, sendo 312 matrizes e 30 varrões oriundos de sete granjas de ciclo completo e distribuídas em cinco municípios do Estado de Alagoas. O diagnóstico sorológico da infecção por C. abortus foi realizado através da microtécnica de Fixação do Complemento (RFC). A análise dos fatores de risco foi realizada por meio da aplicação de questionários investigativos, constituídos por perguntas objetivas referentes ao criador, às características gerais da propriedade, ao manejo produtivo, reprodutivo e sanitário. Observou-se prevalência de 10,5% (36/342) de suínos soropositivos para a infecção por C. abortus, com 85,8% das granjas analisadas com animais positivos. As variáveis que demonstraram associação significativa foram: utilização de bebedouros comuns para jovens e adultos (p=0,024;OR=10,83; IC=1,36–86,03) e método de cobertura de monta natural associada à inseminação artificial (p=0,05; OR=7,62; IC=1,00-58,31). Relata-se a primeira ocorrência de anticorpos anti-C. abortus em suínos no Brasil. Fatores como a introdução de reprodutores nos plantéis e a forma de fornecimento de água foram evidenciados como facilitadores da infecção das matrizes neste estudo. Dessa forma, medidas de controle da infecção devem ser enfocadas nesse aspecto para evitar a disseminação do agente nas granjas suinícolas e em outros plantéis da região.

The objective of this study was to determine the prevalence and identify risk factors associated with Chlamydophila abortus infection in commercial swine farms on the state of Alagoas, Brazil. To compose the study sample 342 pigs were used, with 312 sows and 30 boars from seven swine farms and distributed in five districts of the Alagoas. The serological diagnosis of infection by C. abortus was performed by fixation of complement microtechnique (RFC). The analysis of risk factors were performed by the application of research questionnaires, consisting of objective questions relating to the designer, the general characteristics of the property, the production, reproductive and health management. I was observed a prevalence of 10.5% (36/342) of pigs seropositive for infection by C. abortus with 85.8% of farms with positive animals analyzed. The variables that showed significant association were use of common drinker for young and adults pigs (p=0.024, OR=10.83, CI=1.36-86.03) and associated the natural mount with artificial insemination (p=0.05, OR=7.62, CI=1.00-58.31). This work reports the first occurrence of anti-C. abortusin pigs in Brazil. Factors as the introduction of boars in herds and the form of water supply were seen as facilitators of infection on sows in this study. Thus measures of infection control should be focused on this aspect to prevent the spread of the agent in pig farms and other herds in the region.

Seroprevalence of antibodies to chlamydophila abortus in ovine in the state of Alagoas, Brazil

The goal of this study was to perform a seroepidemiological investigation and to identify risk factors associated with infection of Chlamydophila abortus of sheep herds in the Brazilian state of Alagoas. The study was conducted with samples of 274 ewes with ages equal to or higher than 24 months in 25 herds and in 23 towns located in three regions of the state (Sertão, Agreste and Eastern Alagoas). Anti-C. abortus antibodies were detected using the microcomplement fixation test. The risk factors, were determined based on questionnaires consisting of objective questions, about the farmer and general characteristics of the herd like size, sanitary situation and reproductive management. Among 274 sera samples analyzed for C. abortus, 59 (21.5 percent) were positive with titers >32, 187 (68.3 percent) negative and 28 (10.2 percent) suspect with titers >16. In the 23 towns studied, 20 had positive animals. Among herds 21 (77.7 percent) of had positive animals. The only variable which appeared to be significant in the multivariate analysis was the region, and Sertão was the most significant (p<0.001; OR=3.48; T.I. 1.79 - 6.76). Results indicate that infection by Chlamydophila abortus is widespread on sheep farms in the State of Alagoas. Others studies, however, have to be conducted to isolate the agent in order to confirm the role of the bacteria is reproductive disturbances in sheeps. In addition to that, control and prophylactic measures along with health promoting programs have to be encouraged on the studied farms so that infection reates are reduced.

Seroprevalence of antibodies to Chlamydophila abortus in Ovine in the State of Alagoas, Brazil.

The goal of this study was to perform a seroepidemiological investigation and to identify risk factors associated with infection of Chlamydophila abortus of sheep herds in the Brazilian state of Alagoas. The study was conducted with samples of 274 ewes with ages equal to or higher than 24 months in 25 herds and in 23 towns located in three regions of the state (Sertão, Agreste and Eastern Alagoas). Anti-C. abortus antibodies were detected using the microcomplement fixation test. The risk factors, were determined based on questionnaires consisting of objective questions, about the farmer and general characteristics of the herd like size, sanitary situation and reproductive management. Among 274 sera samples analyzed for C. abortus, 59 (21.5%) were positive with titers ≥32, 187 (68.3%) negative and 28 (10.2%) suspect with titers ≥16. In the 23 towns studied, 20 had positive animals. Among herds 21 (77.7%) of had positive animals. The only variable which appeared to be significant in the multivariate analysis was the region, and Sertão was the most significant (p<0.001; OR=3.48; T.I. 1.79 - 6.76). Results indicate that infection by Chlamydophila abortus is widespread on sheep farms in the State of Alagoas. Others studies, however, have to be conducted to isolate the agent in order to confirm the role of the bacteria is reproductive disturbances in sheeps. In addition to that, control and prophylactic measures along with health promoting programs have to be encouraged on the studied farms so that infection reates are reduced.

Ocorrência e fatores de risco para Chlamydophilaabortus em ovinos e caprinos no estado de Pernambuco: [revisão] / Occurrence and risk factors for Chlamydophilaabortus infection in sheep and goats in Pernambuco: [revision]

Objetivou-se com este estudo identificar os fatores de risco associados à infecção por Chlamydophila abortus em caprinos e ovinos nas regiões do Litoral/ Zonada Mata e Agreste do Estado de Pernambuco. Foram colhidas 290 amostras de soros para pesquisa de anticorpos contra clamídia em 12 propriedades. Para identificar os fatores de risco foram aplicados questionários junto aos proprietários. A freqüência de animais soro-reagentes nos rebanhos estudados foi de 10,3%, sendo 12,0% para caprinos e 8,1% para ovinos, identificando-se 11/12 (91,6%) focos da infecção. Registra-se a primeira ocorrência de anticorpos anti-C. abortus em caprinos no estado de Pernambuco e em ovinos no Brasil. Os fatores derisco associados à infecção em caprinos foram a raça (OR=9,10) e o manejo intensivo (OR=6,41) e para ovinosnão foram encontradas associações significativas para nenhum fator analisado. Concluiu-se que a infecção porChlamydophila sp. encontra-se disseminada em criações de caprinos e ovinos da Zona da Mata e Agreste do Estado de Pernambuco. Medidas de controle devem ser implantadas nos criatórios estudados, enfocando principalmente os fatores de risco identificados neste estudo para reduzir a possibilidade de infecção por este agente.

The study aimed to identify risk factors associated with Chlamydophila abortus infection in sheep and goats from the Litoral/Zona da Mata and Agreste region of Pernambuco state, Brazil. Serum samples (n=290) were analyzed to detect Chlamydophila spp. antibodies in 12 farms. Questionnaires were applied to identify risk factors. Frequency ofserum-reactive animals were 10.3% (12.0% in ewes and 8.1% in goats) and 1/12 (91.6%) infection focuses were identified. This is the first report on anti-Chlamydophhila abortus antibodies in goats and sheep in Pernambuco and Brazil, respectively. Risk factorsassociated with goat infection were breed (OR=9.10) and management (OR=6.41). No significant associations in any of the analyzed factors were found for sheep. In summary, Chlamydophila sp. infection is disseminated in sheep and goat herds in the region. Control measures should be established, focusing primarily risk factor identified in this study, to reduce the possibility of infection by the agent.

Prevalence and risk factors for anti-Toxoplasma gondii antibodies in goats of the Seridó Oriental microregion, Rio Grande do Norte state, Northeast region of Brazil.

The prevalence and risk factors for anti-Toxoplasma gondii antibodies were investigated in goats of the Seridó Oriental microregion, Rio Grande do Norte state, Northeast region of Brazil. Three hundred and sixty-six blood samples from goats collected by jugular venopuncture were used. For the serologic diagnosis of Toxoplasma gondii infection, the indirect fluorescent-antibody test (IFAT) with cut-off value 1:64 was carried out. The prevalence of anti-T. gondii antibodies was 30.6% [95% CI=25.9-35.6%] with titers ranging from 1:64 to 1:16,384. The multivariate logistic regression analysis showed that the risk factors associated to anti-T. gondii antibodies were presence of cats in the herd, extensive/semi-intensive management systems and lack of mineral supplementation.

Detection of Brucella abortus DNA in illegal cheese from São Paulo and Minas Gerais and differentiation of B19 vaccinal strain by means of the polymerase chain reaction (PCR)

A total of 192 samples of illegal cheese from different regions of the states of São Paulo and Minas Gerais, Brazil, were analyzed for the isolation and detection of Brucella spp. DNA by means of microbiological culture and polymerase chain reaction (PCR), respectively. Samples that yielded positive results were submitted to the analysis of the occurrence of Brucella abortus (biovars 1, 2 e 4), as well as to the differentiation of DNA in B19 vaccinal strain or Brucella abortus field strain using PCR. Although the microorganism was not isolated from any sample, PCR detected 37 positive samples (19.27 percent) using genus-specific primers. From these, all (100 percent) were Brucella abortus. Differentiation of the strain showed that 30/37 samples (81.08 percent) were vaccinal strain B19 and seven (18.92 percent) were Brucella abortus field strains. Results showed that diagnostic sensitivity of PCR was greater than that of microbiological culture. The standardization of the reaction for the differentiation of vaccinal and field strains enabled the analysis of all samples positive for Brucella abortus. It is, therefore, a reliable method, also applicable to natural infections caused by the microrganism.

Foram analisadas 192 amostras de queijo clandestinas provenientes de várias regiões do Estado de São Paulo e Minas Gerais, Brasil, para isolamento e detecção de DNA de Brucella spp. através das técnicas de cultivo microbiológico e reação da polimerase em cadeia (PCR), respectivamente. Para as amostras positivas foi pesquisada a ocorrência da espécie Brucella abortus (biovares 1, 2 e 4), além da diferenciação do DNA em cepa vacinal B19 ou de campo por PCR. Não foi possível isolar o microrganismo de nenhuma amostra, porém, na PCR, 37 amostras (19,27 por cento) foram positivas na reação com primers gênero específicos e destas, todas (100 por cento) foram comprovadas como sendo Brucella abortus. A diferenciação da cepa revelou que 30/37 amostras (81,08 por cento) eram cepa vacinal B19 e sete (18,92 por cento) eram cepas de Brucella abortus de campo. Os resultados mostraram uma maior sensibilidade diagnóstica da PCR em relação ao cultivo microbiológico, e a padronização da reação de diferenciação da cepa em vacinal ou campo permitiu que todas as amostras positivas para Brucella abortus fossem analisadas, sendo uma metodologia confiável e aplicável a infecções naturais pelo microrganismo.

Molecular subtyping of Campylobacter jejuni subsp. jejuni strains isolated from different animal species in the state of São Paulo, Brazil

O objetivo do presente trabalho foi caracterizar geneticamente estirpes de Campylobacter jejuni subsp. jejuni isoladas de humanos e de diferentes origens animais (bovinas, suínas, cães, primatas, javalis, suínos e aves de corte). Um total de 828 amostras (fezes, carcaças, fetos abortados e útero histerectomizado) foram analisadas por métodos de rotina bacteriológica e 36 estirpes de C. jejuni foram isoladas. Trinta estirpes de origem fecal humana foram obtidas de laboratórios de análises clínicas da cidade de São Paulo. As 66 estirpes de C. jejuni isoladas foram submetidas à caracterização genética. Oligonucleotídeos baseados no gene fla A foram usados na reação de polimerase em cadeia (PCR) e amplificou um fragmento de 702 pb. Os produtos obtidos pela PCR foram avaliados pelas técnicas de seqüenciamento e análise genealógica. Análise da variabilidade genética das 66 estirpes revelou 44 diferentes subtipos de C. jejuni. Um subtipo de origem humana apresentou seqüência idêntica à de C. jejuni depositada no GenBank (GENBANK acesso número AF050186). A subtipagem das estirpes de C. jejuni baseadas no seqüenciamento da região variável do gene fla A e na análise do alinhamento das seqüências pelo método da Máxima Parcimônia, mostraram-se altamente discriminatórios fornecendo melhores condições para a correta diferenciação entre estirpes originárias de surto e as isoladas esporadicamente. Este foi o primeiro estudo de subtipagem molecular de estirpes de C. jejuni de origem humana e animal utilizando a técnica do seqüenciamento com análise genealógica realizado no Estado de São Paulo, Brasil.

Leptospira spp detection by Polymerase Chain Reaction (PCR) in clinical samples of captive black-capped Capuchin monkey (Cebus apella)

Leptospirosis is a widely distributed zoonosis that affects domestic and wild animals, and that has the man as the end point of its epidemiological chain. Leptospirosis diagnosis in primates is more difficult than in other animal species, as clinical signs and lesions are less evident and antibody response is detected only for short periods. The aim of this article was to describe the detection of Leptospira spp using polymerase chain reaction (PCR), in clinical samples from one captive black-capped Capuchin monkey (Cebus apella), which presented characteristics compatible with leptospirosis (jaundice and haemorrhagic kdney) in the macroscopic post-mortem examination. A friable kidney fragment and urine sample were cultured and submitted to experimental inoculation in guinea pigs and PCR using genus specific primer pair targeting the 16S rRNA region from Leptospira interrogans serovar canicola. Isolation of the agent was negative both in culture and experimental inoculation. The PCR amplification of the clinical samples showed a 330 pb amplified fragment that corresponds to the Leptospira genus. Based on these results PCR was considered an important tool for leptospira detection in nonhumam primates, more sensitive and specific than other techniques, especially considering that the viability of the pathogen was not possible. These advantages enable the detection of the leptospiras in urine and kidney, even when autolysed, frozen or badly conserved, which prevented the isolation and experimental inoculation from positive results.

Characterization of Aujeszky's disease virus isolates from South and Southeast Brazil by RFLP analysis

O DNA genômico de 30 amostras do vírus da doença de Aujeszky (VDA), isoladas nas regiões Sul e Sudeste do Brasil no período de 1982 a 1996, foram caracterizadas por análise do perfil de restrição utilizando a enzima BamHI. Vinte e sete amostras foram isoladas de suínos, uma de bovino, uma de felino e uma de canino. Através de uma sistemática previamente descrita, as 30 amostras de VDA foram classificadas como pertencentes ao tipo genômico I (n = 2) e tipo genômico II (n = 28). O tipo genômico III não foi observado. Neste primeiro estudo de caracterização genômica de isolados de VDA do Brasil, foi possível demonstrar a ocorrência dos tipos genômicos I e II, com marcante predomínio do tipo II.