RESUMO 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.

ABSTRACT 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.

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.

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.

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.

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%. 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.

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.

Objetivou-se com este estudo identificar os fatores de risco associados à infecção por Chlamydophila abortus em caprinos e ovinos nas regiões do Litoral/ Zona da 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 de risco associados à infecção em caprinos foram a raça (OR=9,10) e o manejo intensivo (OR=6,41) e para ovinos não foram encontradas associações significativas para nenhum fator analisado. Concluiu-se que a infecção por Chlamydophila 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 of serum-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 factors associated 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.

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.

The objective of the present trial was to characterize genetically strains of Campylobacter jejuni subsp. jejuni isolated from humans and several animal sources (bovines, swine, dogs, primates, wild boars and poultry). A total of 828 different animal samples (feces, carcass, aborted fetus and hysterectomized uterus) were analysed by means of routine bacteriological methods, and 36 C. jejuni strains were isolated. Thirty strains of human fecal origin were obtained in clinical analysis laboratories in the city of São Paulo. The 66 C. jejuni strains isolated were submitted to genetic characterization. Primers based on fla A gene were used in a polymerase chain reaction (PCR) and amplified a fragment of the 702 bp. PCR products were evaluated by means of sequencing and genealogic analysis. Genetic variability analysis of 66 strains showed 44 different subtypes of C. jejuni. One subtype was identical to a C. jejuni strain of human origin with the sequence in the GenBank (GENBANK accession number AF050186). Subtyping analysis of C. jejuni strains based on sequencing of the fla A gene variable region and analysis of sequence alignment by the Maximum Parsimony method showed to be highly discriminatory, providing the best conditions to differentiate strains involved in outbreaks from those sporadically isolated. This is the first study of molecular subtyping analysis of human and animal C. jejuni strains using sequencing technique and genealogic analysis in the state of São Paulo, Brazil.

A leptospirose é uma zoonose de ampla distribuição geográfica que acomete os animais domésticos e silvestres, tendo o homem como ponto final da cadeia epidemiológica. O diagnóstico da leptospirose em primatas é dificultado em relação a outras espécies animais, porque os sinais clínicos e lesões são menos evidentes e a resposta de anticorpos é detectada apenas em curtos períodos. O presente trabalho tem por objetivo descrever a detecção de Leptospira spp empregando-se a Reação em Cadeia pela Polimerase (PCR) em amostras clínicas provenientes de um macaco prego (Cebus apella) criado em cativeiro, que apresentou ao exame macroscópico realizado durante a necrópsia, características sugestivas de leptospirose (icterícia e rins hemorrágicos). Amostras de urina e fragmento de rim friável foram submetidas às técnicas cultivo e inoculação experimental em cobaias e PCR empregando-se primers gênero específicos da seqüência do gene 16S rRNA de Leptospira interrogans sorovar canicola. Na PCR as amostras clínicas revelaram um fragmento amplificado de 330 pb correspondente ao gênero Leptospira. O isolamento do agente foi negativo nas demais técnicas empregadas. Pelos resultados obtidos, verifica-se que a técnica de PCR representou uma importante ferramenta de detecção de leptospiras em primatas não-humanos, principalmente considerando-se que não foi necessária a viabilidade do patógeno, permitindo a detecção do agente em amostras autolisadas, congeladas ou amostras mal conservadas, que prejudicaram o isolamento e a inoculação experimental.

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.

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.

The genomic DNA of thirty strains of Aujeszky's disease virus (ADV) isolated in the South and Southeast regions of Brazil from 1982 to 1996 were characterized by restriction endonuclease analysis with BamHI. Twenty seven strains were isolated from pigs, 1 from cattle, 1 from cat and 1 from dog. Using a systematization previously described, the 30 ADV strains could be classified as genomic types I (n = 2) and II (n = 28). Genomic type III was not observed. In this first study of genomic type characterization of brazilian ADV strains, we could demonstrate the occurence in Brazil of the genomic types I and II, with a large predominance of genomic type II.