Identification of clostridium perfringens and Salmonella spp. in swine through PCR

Background: The presence Clostridium spp. and Salmonella spp. is the main concern in swine sector, since they interfere in the microbiological quality of meat and can be responsible for several diseases, presenting occurrences such as intestinal infections and enterotoxaemia. It is necessary to improve production and quality of meat and by-products swine ensuring good quality origin of products marketed. Considering these aspects, the objective of the present study was to assess the presence of Clostridium perfringens and Salmonella spp. in pigs destined for consumption, with bacterial counts and identification using PCR technique.Materials, Methods & Results: The collections were made at slaughterhouse and confinement, with bacterial count by the pour-plate technique with Reinforced Clostridial Agar, and the identification by PCR with cpa, cpb, IA, cpe, etx and cpb-2 genes for C. perfringens; and invA and fliC for Salmonella spp. The scores presented average values of 6.04 x 104 CFU/mL for confinement, and 1.8 x 104 CFU/mL in the slaughterhouse. The means counts between the confinement and slaughterhouse groups showed significant difference (P < 0.01). The samples taken at the slaughterhouse were from several farms at random, and allowed the appraisal of hygienic conditions, since animals of confinement remained sanitized and the accumulation of feces on floor was avoided. The PCR technique presented positive results for C. perfringens alpha toxin (cpa) in 25% of the confinement samples, and 46.4% in the slaughterhouse. Salmonella spp. was identified in 9.5% of confinement and 21.4% of the slaughterhouse. Discussion: The presence of C. perfringens and Salmonella spp. is directly related to losses in the entire system of creation of pigs, since infected animals are susceptible to severe diseases, or without clinical signs can exhibit apathy and loss of appetite making weight gain difficult and causing deficiency of the immune system.[...](AU)

Identification of clostridium perfringens and Salmonella spp. in swine through PCR

Background: The presence Clostridium spp. and Salmonella spp. is the main concern in swine sector, since they interfere in the microbiological quality of meat and can be responsible for several diseases, presenting occurrences such as intestinal infections and enterotoxaemia. It is necessary to improve production and quality of meat and by-products swine ensuring good quality origin of products marketed. Considering these aspects, the objective of the present study was to assess the presence of Clostridium perfringens and Salmonella spp. in pigs destined for consumption, with bacterial counts and identification using PCR technique.Materials, Methods & Results: The collections were made at slaughterhouse and confinement, with bacterial count by the pour-plate technique with Reinforced Clostridial Agar, and the identification by PCR with cpa, cpb, IA, cpe, etx and cpb-2 genes for C. perfringens; and invA and fliC for Salmonella spp. The scores presented average values of 6.04 x 104 CFU/mL for confinement, and 1.8 x 104 CFU/mL in the slaughterhouse. The means counts between the confinement and slaughterhouse groups showed significant difference (P < 0.01). The samples taken at the slaughterhouse were from several farms at random, and allowed the appraisal of hygienic conditions, since animals of confinement remained sanitized and the accumulation of feces on floor was avoided. The PCR technique presented positive results for C. perfringens alpha toxin (cpa) in 25% of the confinement samples, and 46.4% in the slaughterhouse. Salmonella spp. was identified in 9.5% of confinement and 21.4% of the slaughterhouse. Discussion: The presence of C. perfringens and Salmonella spp. is directly related to losses in the entire system of creation of pigs, since infected animals are susceptible to severe diseases, or without clinical signs can exhibit apathy and loss of appetite making weight gain difficult and causing deficiency of the immune system.[...]

Kinetics of epsilon antitoxin antibodies in different strategies for active immunization of lambs against enterotoxaemia / Cinética dos anticorpos antitoxina epsilon em diferentes estratégias de imunização ativa em cordeiros contra a enterotoxemia

Enterotoxaemia, a common disease that affects domestic small ruminants, is mainly caused by the epsilon toxin of Clostridium perfringens type D. The present study tested four distinct immunization protocols to evaluate humoral response in lambs, a progeny of non-vaccinated sheep during gestation. Twenty-four lambs were randomly allocated into four groups according to age (7, 15, 30 and 45 days), receiving the first dose of epsilon toxoid commercial vaccine against clostridiosis with booster after 30 days post vaccination. Indirect ELISA was performed after the first vaccine dose and booster to evaluate the immune response of the lambs. Results showed that for the four protocols tested all lambs presented serum title considered protective (≥0.2UI/ml epsilon antitoxin antibodies) and also showed that the anticipation of primovaccination of lambs against enterotoxaemia conferred serum title considered protective allowing the optimization of mass vaccination of lambs.(AU)

Enterotoxemia, uma das mais comuns enfermidades que acomete os pequenos ruminantes domésticos, é causada principalmente pela toxina épsilon de Clostridium perfringens tipo D. O presente estudo avaliou a resposta humoral conferida por quatro protocolos distintos de primovacinação na progênie de ovelhas não vacinadas durante a gestação. Vinte e quatro cordeiros foram aleatoriamente divididos em quatro grupos de acordo com a idade (dias) que receberam a primeira dose da vacina comercial contra clostridiose contendo toxóide epsilon na sua formulação. Todos os cordeiros foram vacinados aos 7, 15, 30 ou 45 dias de idade e receberam um reforço da dose 30 dias após a vacinação. A avaliação sorológica dos cordeiros pelo teste de ELISA indireto foi realizada por ocasião da administração da primeira dose da vacina. Os resultados elucidaram não haver comprometimento da resposta imune de cordeiros vacinados tanto aos 7, 15, 30 ou 45 dias de idade associada ao reforço da dose 30 dias após, demonstrando assim que a antecipação da primeira vacinação conferiu proteção aos cordeiros contra a enterotoxemia, permitindo otimizar o planejamento da vacinação em massa dos cordeiros.(AU)

Cinética dos anticorpos de origem colostral contra a toxina épsilon de Clostridium perfringens tipo D em cordeiros / Kinetics of colostral antibodies against epsilon toxin produced by Clostridium perfringens type D in lambs

Enterotoxemia, uma das mais importantes enfermidades que acomete os pequenos ruminantes domésticos, é causada principalmente pela toxina épsilon de Clostridium perfringens tipo D. O presente estudo avaliou a cinética de anticorpos colostrais antitoxina épsilon em cordeiros nascidos de ovelhas submetidas a dois diferentes tipos de manejo sanitário. Um grupo de ovelhas prenhes (n=6) foi vacinado com uma dose única de vacina comercial polivalente contra clostridioses contendo toxóide épsilon na sua formulação cerca de 30 dias antes da data prevista para a parição. Outro grupo de ovelhas (n=6) de mesma idade gestacional não foi vacinado. Imediatamente após o parto, antes da ingestão do colostro, foram colhidas amostras sanguíneas dos respectivos cordeiros, bem como aos 30 e 60 dias de idade e submetidas à avaliação sorológica pelo teste de ELISA indireto. Os resultados encontrados permitem concluir que a vacinação de ovelhas prenhes 30 dias antes do parto contra a enterotoxemia causada pela toxina épsilon, com dose única de produto comercial, induz imunidade passiva em níveis considerados protetores (>0,5UI/ml) aos cordeiros por, no mínimo, 60 dias de idade.(AU)

Enterotoxemia, a disease that affect small ruminants, is caused mainly by the epsilon toxin from Clostridium perfringens type D. This study evaluated the kinetics of epsilon antitoxin colostral antibodies in lambs born to ewes submitted to two different types of health management. A group of pregnant ewes (n=6) was vaccinated with a single dose of commercial vaccine against polyvalent clostridial toxoid containing epsilon in its formulation some 30 days before the expected date of birth. Another group of ewes (n=6) of the same gestational age were not vaccinated. Immediately after birth, before intake of colostrum, blood samples were collected from their lambs as well as 30 and 60 days of age for serum evaluation by ELISA. The results allow to conclude that vaccination of pregnant ewes 30 days before parturition allowed the transfer of colostral antibodies specific enough to guarantee the immunity considered protective (0,5UI/ml) of the newborn lamb against enterotoxemia until its entry into the feedlot, with 60 days of age.(AU)

Coccidiosis, Paratuberculosis and Enterotoxaemia in Saudi Goats

During the year 2010, 66 goats suffering from mucoid to hemorrhagic diarrhea were submitted to the Pathology Division, Diagnostic Veterinary Laboratory, Jeddah, Saudi Arabia. After postmortem examination, tissue specimens were obtained from liver, lungs, heart, brain, stomach, small and large intestine and mesenteric lymph nodes for microbiological and histopathological examinations. Data from Microbiology Laboratory revealed that goats had intestinal coccidiosis (34 cases), paratuberculosis (9 cases), enterotoxaemia (22 cases) and mycotic gastritis (1 case). In the present report, we focused on the pathological pictures of the above mentioned diseases in examined goats.(AU)

Enterite necrótica aviária

Avian Necrotic Enteritis is an acute enterotoxaemia that appears suddenly producing rapid deaths, affecting mainly young animals. Although its negative impact in poultry production is already known, factors related to etiology, pathogenesis and control of this important disease need better clarifications. For a long time its control was based on the use of antibiotics in poultry feed, whose the use was banned by several consumer markets, requiring the development of new control strategies. Informations on the etiology, epizootiology, pathogenesis, diagnosis and control are reviewed, emphasizing the role of probiotics and vaccines as control alternatives.

A Enterite Necrótica Aviária (ENA) é uma enterotoxemia aguda que aparece subitamente e provoca morte rápida, afetando principalmente animais jovens. Embora seu impacto negativo na produção, devido ao aumento da conversão alimentar e da condenação de carcaças seja já conhecido, questões relacionadas à etiologia, à patogenia e ao controle desta importante enfermidade necessitam de maiores esclarecimentos. Nos últimos anos, o controle da ENA baseou-se na aplicação de antibióticos na ração animal, prática banida pelo mercado consumidor, que exigiu o desenvolvimento de novas estratégias de controle. Esta revisão aborda informações sobre a etiologia, a epizootiologia, a patogenia, o diagnóstico e o controle da doença, em especial a utilização de probióticos e vacinas como alternativas de controle da ENA.

Enterite necrótica aviária

Avian Necrotic Enteritis is an acute enterotoxaemia that appears suddenly producing rapid deaths, affecting mainly young animals. Although its negative impact in poultry production is already known, factors related to etiology, pathogenesis and control of this important disease need better clarifications. For a long time its control was based on the use of antibiotics in poultry feed, whose the use was banned by several consumer markets, requiring the development of new control strategies. Informations on the etiology, epizootiology, pathogenesis, diagnosis and control are reviewed, emphasizing the role of probiotics and vaccines as control alternatives.

A Enterite Necrótica Aviária (ENA) é uma enterotoxemia aguda que aparece subitamente e provoca morte rápida, afetando principalmente animais jovens. Embora seu impacto negativo na produção, devido ao aumento da conversão alimentar e da condenação de carcaças seja já conhecido, questões relacionadas à etiologia, à patogenia e ao controle desta importante enfermidade necessitam de maiores esclarecimentos. Nos últimos anos, o controle da ENA baseou-se na aplicação de antibióticos na ração animal, prática banida pelo mercado consumidor, que exigiu o desenvolvimento de novas estratégias de controle. Esta revisão aborda informações sobre a etiologia, a epizootiologia, a patogenia, o diagnóstico e o controle da doença, em especial a utilização de probióticos e vacinas como alternativas de controle da ENA.

Enterotoxemia em bovino / Bovine enterotoxaemia

This report describes a case of bovine enterotoxaemia in Morro da Garça, Minas Gerais, Brazil. Clostridium perfringens type D was isolated in pure culture and was characterized by biochemical reactions and PCR. By the mouse neutralization test, the presence of epsilon toxin from intestinal content was detected.(AU)

Diarréia em leitões lactentes por Clostridium perfringens tipo A em granjas tecnificadas nos estados de Minas Gerais e São Paulo / Clostridium perfringens type A diarrhea in suckling piglets in industrial swine farms in the states of Minas Gerais and São Paulo

Diarrhea in suckling piglets caused by Clostridium perfringens type A was diagnosed in industrial (technified) swine farms of the states of Minas Gerais and São Paulo (Brazil), based on isolation and identification of bacterium by biochemical tests, detection of alpha toxin in animal bioassays, and PCR. This seems to be the first report of clostridial enterotoxaemia in piglets by C. perfringens type A in Brazil and allowed specific procedures to control the disease.(AU)

Coccidiosis, paratuberculosis and enterotoxaemia in Saudi goats

During the year 2010, 66 goats suffering from mucoid to hemorrhagic diarrhea were submitted to the Pathology Division, Diagnostic Veterinary Laboratory, Jeddah, Saudi Arabia. After postmortem examination, tissue specimens were obtained from liver, lungs, heart, brain, stomach, small and large intestine and mesenteric lymph nodes for microbiological and histopathological examinations. Data from Microbiology Laboratory revealed that goats had intestinal coccidiosis (34 cases), paratuberculosis (9 cases), enterotoxaemia (22 cases) and mycotic gastritis (1 case). In the present report, we focused on the pathological pictures of the above mentioned diseases in examined goats.

Sudden death of goats due to enterotoxaemia / Morte súbita de caprinos por enterotoxemia

An occurrence of sudden death in goats is presented. Most of the death were without symptons but some animals exhibited diarrhoea, incoordination of movements and, before their death, spelted strong cries. The isolation of anaerobic Gram-positive rod, biochemically identified as Clostridium perfringens was possible in intestines, reticulum and food samples. Filtrates of these organs revelated a termolabile and high lethal toxin when inoculated into mice. Macroscopically it was seen hemorrhages in the intestines mucosa and stomachs filled up with food. Those aspects associated to the laboratory findings led to the diagnostic of enterotoxaemia.

Descreve-se a ocorrência de morte súbita observada em uma criação de caprinos. A maioria das mortes ocorreu sem qualquer manifestação sintomática prévia; entretanto, em alguns casos observou-se incoordenação motora, eliminação de cíbalas envoltas em fragmentos de mucosa intestinal e diarréia. Nos momentos que antecediam à morte, alguns animais emitiam fortes berros. O isolamento de um bacilo anaeróbio, Gram-positivo e bioquimicamente identificado como Clostridium perfríngens, foi possível a partir de amostras de materiais coletadas em diferentes porções dos intestinos, retículo e também da ração. Nos filtrados obtidos dos conteúdos dos órgãos citados, evidenciou-se a presença, em alta dosagem, de uma toxina termolábil e letal para camundongos. À necrópsia, observaram-se compartimentos gástricos plenos de alimento e hemorragias na mucosa intestinal. Estas constatações, associadas aos resultados laboratoriais, possibilitaram concluir tratar-se de um quadro de enterotoxemia.

Enterotoxemia em caprinos no Rio Grande do Sul / Caprine enterotoxaemia in Rio Grande do Sul, Brazil

São descritos surtos de enterotoxemia em caprinos em cinco propriedades no Estado do Rio Grande do Sul. Os animais afetados eram, normalmente, encontrados mortos ou apresentavam evolução aguda de 2 a 3 horas com acentuada depressão, cólicas abdominais e diarréia profusa com fibrina. Em duas propriedades relataram-se casos com a evolução de até 12 horas. Em treze animais necropsiados observaram-se aumento de líquidos nas cavidades abdominal, torácica e pericárdica, congestão e hiperemia da serosa e mucosa do intestino, conteúdo do cólon líquido com fibrina além de hemorragias de serosa e fibrina. Em um animal constatou-se microangiopatia cerebral caracterizada por acúmulo de material homogêneo e eosinofílico no espaço perivascular. No conteúdo intestinal, colônias com bastonetes morfológica e bioquimicamente sugestivos de Clostridium perfringens foram caracterizadas no estudo bacteriológico. A soroneutralização em camundongos com conteúdo intestinal dos animais afetados, revelou a presença da toxina épsilon. Estes achados evidenciam a enterotoxemia como doença de importância para criação de caprinos no Rio Grande do Sul (AU)

Eficácia de vacina experimental contra a enterotoxemia causada pela toxina épsilon do Clostridium perfringens tipo D em caprinos

A enterotoxemia em caprinos é determinada pelos efeitos da toxina épsilon do Clostridium perfringens tipo D. Esta enfermidade é considerada a principal causa infecciosa de mortalidade nas criações de caprinos e ovinos no Brasil. As medidas profiláticas incluem o manejo alimentar correto e o uso de vacinas com alto poder imunogênico. No presente estudo, foram desenvolvidas e avaliadas, quanto à resposta humoral em caprinos, duas vacinas experimentais; ambas as vacinas contendo 9UllmL de toxóide épsilon em adjuvante aquoso (vacina A) e oleoso (vacina B). Foram utilizados 100 caprinos em duas estratégias (1 e 2) distintas de vacinação, com 50 animais em cada. Os animais da estratégia 1 receberam a primeira dose de vacina aos sete dias de idade, e os da estratégia 2, aos 45 dias de idade, ambas seguidas de uma dose de reforço 35 dias após. Cada estratégia foi dividida em cinco grupos com 10 animais cada. Os animais do grupo 1 (G1) receberam somente duas doses da vacina A, e os do grupo 2 (G2) receberam quatro doses dessa mesma vacina. Os animais dos grupos 3 (G3) e 4 (G4) receberam duas ou quatro doses, respectivamente, da vacina B. As doses de reforço foram feitas aos 150 e 270 dias após o início do experimento nos animais do G2 e G4. Os animais do grupo controle (C) não foram vacinados. As amostras de soro sangüíneo foram colhidas nos dias zero, 15,35,50,65,90 e depois a cada 30 dias até os 390 dias. A ocorrência de reações nos locais de aplicação das vacinas foi avaliada pelo registro da formação de granulomas elou abscessos. A quantificação dos anticorpos antitoxina épsilon foi avaliada pela técnica de ELlSA Indireto. Os caprinos pertencentes a todos os grupos foram desafiados experimentalmente aos 105 dias de idade, por meio de infusão intraduodenal de cultura pura de C. perfringens tipo D contendo 9.000 DLsoIKg de peso vivo de toxina épsilon

Enterotoxaemia in goats is believed to be caused by the epsilon toxin of Clostridium perfringens type D. This disease is considered the most important infectious cause of death in ovine and caprine production systems in Brazil. Prophylactic measures against this disease include proper feeding management and use of adequate vaccines of high immunogenic power. In the present trial, two experimental vaccines were developed and evaluated in relation to humoral responses in goats. Both vaccines contained 9Ul/mL of epsilon toxoid in aqueous (vaccine A) and oily (vaccine B) adjuvant. One hundred goats were used with two different vaccination strategies (1 and 2) with 50 animais each. Vaccination schedule for animais in strategy 1 began at seven days of age, while in strategy 2, it began at 45 days of age; both with one booster dose 35 days after. Each strategy was in tum divided into 5 groups of 10 animais each. Animais in group 1 (G1) only received two doses of vaccine A and those of group 2 (G2) received four doses of the same vaccine. Animais of group 3 (G3) and 4 (G4) received two and four doses, respectively, of vaccine B. Booster doses were given in days 150 and 270 of the beginning of the trial in the group G2 and G4. Animais of the control group (C) were not vaccinated. Samples of blood serum were collected on days zero, 15, 35, 50, 65, 90 and after that, every 30 days until 390 days. The occurrence of reaction in the site of application was evaluated by registry of formation of the granulomas and abscesso Measurement of epsilon antitoxin was performed using an Indirect ELlSA. Goats from ali groups were experimentally challenged at 105 days of age with an intraduodenal infusion of apure culture of type D C. perfringens containing 9,000 DLsoIKg body weight of epsilon toxin