Caracterização de estirpes de Campylobacter coli isoladas de carcaças de ovinos e de efluentes de abatedouro do estado de São Paulo / Characterization of Campylobacter coli strains isolated from the carcasses of sheep and of wastewater of abattoir in the state of São Paulo

Doença bacteriana zoonótica, a campilobacteriose é responsável mundialmente por frequentes casos de gastroenterite humana. Campylobacter spp. apresenta fator de virulência associado à diarreia, denominado toxina citoletal distensiva (CDT), sendo codificado pelos genes do complexo cdt. Os objetivos do presente estudo foram: 1) isolar e identificar estirpes de Campylobacter spp. de 102 suabes de carcaças e 102 suabes retais de ovinos (Ovis aries) e de sete amostras de água dos efluentes, antes e depois do tratamento de desinfecção de abatedouro localizado no estado de São Paulo; e 2) detectar, pela técnica de multiplex-PCR, a presença do complexo de genes cdt. Foram isoladas e identificadas, por métodos fenotípicos e genotípicos, sete estirpes de Campylobacter coli provenientes de 4/102 (3,92%) das amostras de suabes retais, 1/102 (0,98%) de suabes de carcaças e 2/7 (28,5%) das águas dos efluentes. Dos isolados de suabes retais, em 2/7 (28,6%) estirpes foi detectada a presença dos genes cdt. Trata-se do primeiro relato de isolamento de estirpes de Campylobacter coli provenientes de abatedouro de ovinos e das estirpes portadoras do complexo de genes cdt nessa espécie animal no Brasil.

A zoonosis and bacterial disease, campylobacteriosis is responsible for frequent cases of human gastroenteritis worldwide. Campylobacter spp. presents the virulence factor called cytolethal distensive toxine (CDT), responsible for diarrhea and codified by the cdt gene. The aims of this study were: 1) to isolate and identify Campylobacter spp. strains from 102 carcass swabs and 102 rectal swabs of sheep (Ovis aries) and seven samples of wastewater, before and after the disinfection treatment, collected from the abattoir of the state of São Paulo; and 2) to detect the presence of cdt gene complex by Multiplex-PCR in strains of Campylobacter spp. Seven strains of Campylobacter coli were isolated and identified by phenotypic and genotypic methods: 4/102 (3.92%) from rectal swabs, 1/102 (0.98%) from carcass swabs and 2/7 (28.5%) from wastewater. From the rectal swab samples 2/7 (28.6%) strains were detected with the cdt gene. This is the first report on the isolation of Campylobacter coli from sheep abattoir, and of strains carrying the cdt gene complex in this animal species in Brazil.

Detecção de fatores de virulência em estirpes de Campylobacter spp. isoladas de carcaças de suínos abatidos em frigoríficos / Virulence factors in strains of Campylobacter spp. isolated from the carcasses of swine slaughtered in abattoirs

Isolaram-se estirpes de Campylobacter spp. em amostras de carcaças (n=65), fezes (n=65) e linfonodos mesentéricos (n=65) de suínos abatidos em frigoríficos do estado de São Paulo e detectaram, pela técnica da Multiplex-PCR, a presença do complexo de genes cdt, responsáveis pela expressão do fator de virulência da toxina CDT. Do total de 195 amostras de origem suína, Campylobacter spp. foi isolado de 31 (15,9%), sendo 29 (93,6%) de amostras de suabe retal, 1/65 (3,2%) de suabe de carcaça e um (3,2%) de linfonodo. Vinte e oito estirpes de C. coli foram positivas para a detecção dos genes cdt, e três estirpes de C. jejuni foram negativas para a detecção desses genes. Foi detectada, pela primeira vez no estado de São Paulo, a presença dos genes cdt em 100% das estirpes de Campylobacter coli provenientes de suínos abatidos em frigoríficos.

The purposes of this study were to isolate and identify Campylobacter spp. strains from the carcasses (n=65), feces (n=65) and mesenteric lymph nodes (n=65) of swine slaughtered in abattoirs in the State of Sao Paulo and to detect the presence of the cdt gene complex - responsible for the expression of the virulence factor cytolethal distensive toxin - in these Campylobacter spp. strains through Multiplex-PCR. From 195 samples analyzed, Campylobacter spp. was isolated in 31 (15.9%): 29 (93,6%) samples of rectal swab, 1 (3.2%) carcass swab and 1 (3.2%) lymph node sample. The 28 strains of isolated C. coli were positive for CDT toxin genes and the three strains of isolated C. jejuni were negative for these genes. It was also the first time that the cdt gene cluster was detected in strains isolated from swine in the state of São Paulo. These findings indicate swine as a potential spreading source of virulent strains of Campylobacter coli, either for slaughterhouse staff or consumers of carcasses and sub products.

Caracterização molecular e fenotípica de estirpes de Escherichia coli produtoras de shiga-toxina (STEC) não-O157 de fezes e carcaças bovinas / Molecular and phenotypic characterization of shiga toxin producing Escherichia coli (STEC) non-O157 strains from bovine feces and carcass

Foram coletados 100 suabes retais e 100 suabes de carcaças bovinas em matadouros do estado de São Paulo, e um total de 326 estirpes de E. coli foram identificadas, sendo 163 de amostras retais e 163 de amostras de carcaça. Todos os isolados submetidos à PCR para detecção dos genes das toxinas Stx1 e Stx2 foram identificados como não-O157 e fenotipados pelo teste da citotoxicidade em células Vero. Das 26 estirpes que apresentaram apenas o gene stx1, das 56 que apresentaram apenas o gene stx2 e das 30 estirpes que apresentaram ambos os genes, 17 (65,4%), 42 (75%) e 22 (73,3%), respectivamente, foram positivas ao teste de citotoxicidade. Não houve diferença estatística entre os três perfis genéticos e na positividade ao teste de citotoxicidade. Os resultados mostram a alta frequência de expressão dos fatores de virulência das STEC de bovinos.

In the present study 100 rectal and 100 carcass swabs were collected from bovines at slaughterhouses in São Paulo state, and the total of 326 E. coli strains were identified (163 from rectal samples and 163 from carcass samples). All the isolates were submitted to PCR for Stx1 and Stx2 toxin gene detection and all strains were identified as non-O157 and phenotyped by the citotoxicity test in Vero cells. Out of 26 strains that presented only the stx1 gene, 56 that presented only the stx2 gene and 30 that presented both genes, 17 (65.4%), 42 (75%) and 22 (73.3%), respectively, were positive for the citotoxicity test. There was no statistically significant difference among these three toxinotyping profiles and positivity in the citotoxicity test, but the results show high frequency of virulence factor expression of bovine.

Prejuízos causados pela mastite em ovelhas Santa Inês / Damages caused by mastitis in Santa Inês Brazilian sheep breed

A mastite tem causado grandes prejuízos em criações de ovinos para corte no Brasil, onde o número de cordeiros nascidos e desmamados é de enorme importância econômica para o sistema de produção. O objetivo deste trabalho foi quantificar esses prejuízos em uma propriedade com ovinos da raça Santa Inês, em sistema semi-intensivo de produção, no Estado de São Paulo. Foram feitos exames clínicos, o Califórnia Mastitis Test (CMT), e as colheitas de leite para exame microbiológico das ovelhas na segunda semana de parição e no desmame. No mesmo dia das colheitas de amostras de leite, os cordeiros foram pesados, bem como no dia do nascimento. Quarenta e uma ovelhas pariram 55 cordeiros (taxa de natalidade 1,34) na estação de parição de 23/10/2008 a 4/1/2009. Destes, 20 (36%) morreram, 14 (70%) nas primeiras duas semanas de vida, e seis (30%) antes do desmame, que ocorreu em torno de 70 dias. 70% (14) das mortes foram atribuídas à condição de mastite da ovelha, e 30% (6) a outras causas, principalmente pneumonia. O ganho de peso diário do nascimento ao desmame de cordeiros filhos de ovelhas com mastite, em pelo menos uma glândula mamária, foi inferior (P < 0,10) ao de cordeiros filhos de ovelhas sadias. Das amostras positivas no exame microbiológico, 85,8% estavam positivas para bactérias do gênero Staphylococcus.

In meat sheep herds in Brazil, mastitis has provoked many damages, causing mortality and decreasing the numbers of lambs born and weaned, posing heavy economic impacts for the production system. The objective of this study was to quantify the damages caused by mastitis in a Santa Inês breed herd, reared in a semi-intensive production system, in São Paulo State, Brazil. Clinical exams were conducted and the California mastitis test was carried out, and milk samples were taken from the ewes for microbiological assay. The lambs were weighed at birth and on the days when the milk samples were taken. Forty-one ewes gave birth to 55 lambs (birth rate of 1.34) during the parturition season from 23 October 2008 to 4 January 2009. Twenty (36%) of them died, 14 (70%) in the first 2 weeks following birth, and 6 (30%) before weaning, which occurs at about 70 days. A total of 70% (14) of the deaths were imputed to the occurrence of mastitis in the respective ewe, and 30% (6) to other causes, mainly pneumonia. Mastitis decreased the average daily gain of the lambs from birth to weaning (P < 0.10). Most of the positive microbiological samples (85.8%) where positive for Staphylococcus spp.

Prevalência de anticorpos contra os agentes da maedi-visna e clamidofilose em ovinos no município de Uberlândia, MG / Prevalence of antibodies against the agents of maedi-visna and chlamydophilosis in sheep from Uberlândia county, MG, Brazil

Objetivou-se com este estudo realizar o primeiro inquérito soro-epidemiológico para o vírus da maedi-visna e Chlamydophila spp. em 12 rebanhos de ovinos do Município de Uberlândia, MG. Foram utilizadas 334 amostras de soro sanguíneo de ovinos e aplicou-se um inquérito epidemiológico a cada propriedade. Os testes realizados para a pesquisa de anticorpos contra o vírus da maedi-visna e Chlamydophila spp. foram imunodifusão em gel de ágar (IDGA) e reação de fixação do complemento (RFC), respectivamente. Não foram detectados ovinos reagentes para maedi-visna. Verificou-se uma prevalência de 3,3% para Chlamydophila spp., com títulos variando de 32 a 64. Não houve diferença estatística significativa (p > 0,05) para os fatores de risco analisados. Ressalta-se a importância dos sistemas de vigilância epidemiológica para atuar no controle dessas infecções, evitando a introdução do vírus da maedi-visna e uma maior propagação da Chlamydophila spp. neste município.

The aim of this study was to carry out the first investigation into the serological prevalence of maedi-visna virus and Chlamydophila spp. on 12 sheep breeding farms in Uberlândia County, MG, Brazil. A total of 334 blood serum samples were used and an epidemiological survey was completed by each farm. The tests to detect maedi-visna and Chlamydophila spp. antibodies were an agar gel immunodiffusion (AGID) and a complement fixation test (CFT), respectively. None of the sheep were reactive to maedi-visna. The detection of antibodies against Chlamydophila spp. was 3.3%, with titers varying from 32 to 64. There was no statistically significant difference (p > 0.05) in regard to the risk factors analyzed. The importance of epidemiological surveillance systems to aid in the control of these infections is emphasized, in order to avoid the introduction of maedi-visna virus and a wider spread of Chlamydophila spp. in this county.

Genotyping of Clostridium perfringens associated with sudden death in cattle

Toxigenic types of Clostridium perfringens are significant causative agents of enteric disease in domestic animals, although type E is presumably rare, appearing as an uncommon cause of enterotoxemia of lambs, calves and rabbits. We report herein the typing of 23 C. perfringens strains, by the polymerase chain reaction (PCR) technique, isolated from small intestine samples of bovines that have died suddenly, after manifesting or not enteric or neurological disorders. Two strains (8.7 percent) were identified as type E, two (8.7 percent) as type D and the remainder as type A (82.6 percent). Commercial toxoids available in Brazil have no label claims for efficacy against type E-associated enteritis; however, the present study shows the occurrence of this infection. Furthermore, there are no recent reports on Clostridium perfringens typing in the country.(AU)

Identification of Clostridium chauvoei in clinical samples cultures from blackleg cases by means of PCR

C. chauvoei presence was detected by means of polymerase chain reaction (PCR) from supernatant of culture in cooked meat medium of liver, muscle and metatarsian bone marrow samples of seven calves with blackleg symptoms. The isolation under anaerobic conditions of one muscle sample revealed Clostridium perfringens in pure culture.

Foi detectada presença de Clostridium chauvoei pela reação de PCR a partir de cultivo em cooked meat medium de amostras de fígado, músculo e medula óssea metatarsiana de sete bezerros acometidos de carbúnculo sintomático. O isolamento de uma amostra de músculo sob condições anaeróbias revelou Clostridium perfringens em cultura pura.

Isolation and identification of bovine tuberculosis in a Brazilian herd (São Paulo)

Mycobacterium was verified in animals from a Brazilian dairy herd, a total of 42 samples from 30 cows were submitted to culture and the isolated strains were analyzed by two polymerase chain reaction (PCR), the first specific for species belonging to the Mycobacterium complex (MTBC) and the other for differentiating M. tuberculosis from M. bovis. Twenty seven samples (64.3 percent) from 18 animals (60 percent) were positive for mycobacteria by culture, including samples from 15 retrofaryngeal lymphnodes (55.5 percent), 9 prescapular lymphnodes (33.3 percent), 2 lungs (7.4 percent), and 1 liver (3.7 percent). All isolated colonies were confirmed by PCR to contain MTBC organisms, and were identified as M. bovis by the same methodology.

Clostridium perfringens types A and D associated with enterotoxemia in an 18-month-old goat

Postmortem examination of a Boer buck that died peracutely revealed bowel and liver diffusely congested and edematous. Kidney was apparently edematous. Clostridium perfringens type A was isolated from bowel and type D from kidney. Microscopic examination revealed large areas of necrosis in the renal cortex and medulla (pulpy kidney disease), hyperemia and centrilobular necrosis of the liver, necrosis of the small-intestine wall, pulmonary edema and congestion, intense hyperemia of the cerebellum, hyperemia and edema of the brain.

Antibodies against pseudorabies virus in feral swine in southeast Brazil / Anticorpos contra o vírus da doença de Aujeszky em javalis no estado de São Paulo

Serum samples collected from 358 wild boars (Sus scrofa) in breeding farms in São Paulo, southeast Brazil, from 1998 to 2001, were tested for antibodies against pseudorabies virus (PRV) by means of serum neutralization (SN) and enzyme-linked immunobsorbent assay (ELISA). Seropositive animals were detected in three of seven herds analyzed. Overall seroprevalence as assessed by SN was 30.7 percent, ranging from 25.2 percent to 100 percent for the herds that presented seropositive animals. Indirect ELISA detected lower seroprevalence (19.3 percent). Sensitivity and specificity of ELISA were equal to 57.3 percent and 97.6 percent, respectively. Agreement was equal to 85.2 percent (P<0.0001). These results showed that PRV infections occurred in farmed feral swine in southeast Brazil, and affect pseudorabies eradication program.

Soros de 358 javalis (Sus scrofa), criados em sistema de semiconfinamento em propriedades do estado de São Paulo, foram coletados entre 1998 e 2000 e testados para anticorpos contra o vírus da doença de Aujeszky (VDA), pela técnica de soroneutralização (SN) e ensaio imunoenzimático (ELISA). Foram detectados animais soropositivos em três das sete propriedades analisadas. Do total de javalis testados, 30,7 por cento apresentaram anticorpos neutralizantes contra o VDA, com variação de 25,2 por cento a 100 por cento nas propriedades com animais sororreagentes. O ELISA detectou menor número de sororeagentes (19,3 por cento), sendo a sensibilidade e a especificidade 57,3 por cento e 97,6 por cento, respectivamente, e a correlação observada de 85,2 por cento (P<0,0001). Os resultados mostram que a infecção pelo vírus da doença de Aujeszky ocorre em criações de javalis no estado de São Paulo, e compromete o sucesso de um futuro programa de erradicação da doença na região.