Production and purification of Clostridium perfringens type C beta-toxin and IgG and IgY antitoxins.

OBJECTIVES: This study aimed to produce and purify Clostridium perfringens type C beta-toxin, sheep anti-beta toxin immunoglobulin G (IgG) and chicken immunoglobulin Y (IgY). METHODS: Two methods were used for beta-toxin purification: single-step metal affinity chromatography (MAC) using zinc as a chelator and ion exchange chromatography (IEX). The purified and inactivated beta-toxoids were then administered to sheep and chickens in order to produce IgG and IgY. RESULTS: All assays using the IEX failed. In contrast, MAC purified more than 21 mg of toxin per run in a single-step protocol. The purified and inactivated beta-toxoids were then administered to sheep and chickens, and IgG and IgY were purified with a high yield, medium antibody titer of 50 IU/mL, and high avidity (73.2 %). CONCLUSIONS: C. perfringens type C beta-toxin and sheep or chicken anti-beta toxin IgG and IgY antibodies were successfully produced and purified using a simple protocol. This protocol can be used for the production of components used in the diagnosis and research of necrotic enteritis caused by C. perfringens type C, as well as for the evaluation of existing vaccines and the development of new preventive methods against this disease.

Cattle and goats' humoral response to vaccination with Clostridium perfringens type D purified epsilon toxoids.

Herd vaccination is an important preventive measure against enterotoxemia in ruminants. Vaccination in goats should be performed every four months, and recent studies have shown that immunity in cattle lasts for less than one year. One of the mechanisms for increasing the duration of the immune response is to use purified toxoids as immunogens. The aim of the present study was to evaluate the humoral response in cattle and goats after vaccination with purified and semi-purified Clostridium perfringens type D epsilon toxoid. The following three different vaccines were used: vaccine 1 (V1), a semi-purified toxoid adsorbed to aluminum hydroxide; vaccine 2 (V2), a purified toxoid adsorbed to aluminum hydroxide; and vaccine (V3), a purified toxoid adsorbed on chitosan microparticles. Groups of cattle (n = 6-7) and goats (n = 6-7) were vaccinated on days 0 and 30, and serum samples for antitoxin titration were collected every 30 days for one-year post-vaccination. Goats were revaccinated on day 360, and their serum was evaluated on days 367 and 374. The antibody peaks ranged between 6.90 and 11.47 IU/mL in cattle and from 1.11 to 4.40 IU/mL in goats. In cattle administered with the V1 and V2 vaccines, we observed that the antibody titers were maintained above 0.2 IU/mL until the end of the experiment. In goats, V2 elicited long-lasting antibodies, and all animals maintained the protective titers for 210 days after the first dose. In conclusion, the purified toxoid vaccine with aluminum hydroxide adjuvant was able to induce strong and long-lasting humoral responses in both species and could be an alternative for improving the immunization schedule against enterotoxemia in goats and cattle.

Production and purification of Clostridium perfringens type D epsilon toxin and IgY antitoxin.

The aim of this study was to purify Clostridium perfringens type D epsilon toxin and produce and purify anti-epsilon chicken immunoglobulin Y (IgY). A single-step ion exchange chromatography resulted in a high-yield and high-purity toxin, while ion exchange chromatography followed by gel filtration resulted in the highest purity of the toxin, but at a lower yield. Purified and inactivated epsilon toxin were then administered in chickens via four inoculations and IgY was obtained at a high purity and yield, with an antibody titer of 50 IU/mL and high levels of avidity (73.2%). In summary, C. perfringens type D epsilon toxin and chicken anti-epsilon IgY were successfully produced and purified, and may be used for the diagnosis of enterotoxemia caused by the epsilon toxin, as well as in potency tests of existing and future vaccines against enterotoxemia.

Type C botulism in domestic chickens, dogs and black-pencilled marmoset (Callithrix penicillata) in Minas Gerais, Brazil.

Botulism is a well-known intoxication that affects humans and animals. The disease is endemic in cattle in Brazil and recently emerged as an important disease in commercial laying hens and broiler chickens in Europe. Dogs and other animal species can also be affected. Although antitoxins are commonly administered to humans diagnosed with botulism, in animals this is rarely the case and the treatment of botulism is still based only on support therapy. In the present work, we report an outbreak of type C botulism in Brazil that simultaneously affected domestic chickens, dogs and a black-pencilled marmoset (Callithrix penicillata). The successful use of Clostridium botulinum types C and D antitoxin for the treatment of an affected dog is also described.

Comparison of humoral neutralizing antibody response in rabbits, guinea pigs, and cattle vaccinated with epsilon and beta toxoids from Clostridium perfringens and C. botulinum types C and D toxoids.

Rabbits and guinea pigs are used in the official control and validation of clostridial vaccines, but it is unknown whether the antitoxin titers obtained in these animals corroborate with the humoral response in bovine. The objective of the study was to compare the humoral antibody response of guinea pig and rabbits to those obtained in cattle vaccinated with a commercial vaccine containing Clostridium perfringens epsilon and beta, and Clostridium botulinum types C and D toxoids. This study revealed the same level of humoral response in rabbits and cattle for all four toxoids tested, including C. botulinum types C and D toxoids. In contrast, the titers of neutralizing antibodies against C. botulinum type C toxin in guinea pigs differed from those obtained in cattle. Thus, the present work suggests that the potency test for C. botulinum types C in rabbits agrees more with the humoral response in cattle than the potency test in guinea pigs, thereby making it possible to use only rabbits as models in the official control and validations of clostridial vaccines.

Detection of toxins A/B and isolation of Clostridium difficile and Clostridium perfringens from dogs in Minas Gerais, Brazil.

The objective of this study was to detect C. difficile A/B toxins and to isolate strains of C. perfringens and C. difficile from diarrheic and non-diarrheic dogs in Brazil. Stool samples were collected from 57 dogs, 35 of which were apparently healthy, and 22 of which were diarrheic. C. difficile A/B toxins were detected by ELISA, and C. perfringens and C. difficile were identified by multiplex PCR. C. difficile A/B toxins were detected in 21 samples (36.8%). Of these, 16 (76.2%) were from diarrheic dogs, and five (23.8%) were from non-diarrheic dogs. Twelve C. difficile strains (21.1%) were isolated, of which ten were A(+)B(+) and two were A(-)B(-). All non-toxigenic strains were isolated from non-diarrheic animals. The binary toxin gene cdtB was found in one strain, which was A(+)B(+) and was derived from a non-diarrheic dog. C. perfringens strains were isolated from 40 samples (70.2%). Of these, 18 (45%) were from the diarrheic group, and 22 (55%) belonged to the non-diarrheic group. All isolates were classified as C. perfringens type A and there was an association between the detection of the cpe gene and the presence of diarrhea. Interestingly, ten strains (25%) were positive for the presence of the cpb2 gene. The high rate of detection of the A/B toxins in non-diarrheic dogs suggests the occurrence of subclinical disease in dogs or carriage of its toxins without disease. More studies are needed to elucidate the epidemiology of C. difficile and C. perfringens in dogs and to better our understanding of C. difficile as a zoonotic agent. This is the first study to report the binary toxin gene in C. difficile strains isolated from dogs in Brazil.

Spoligotyping and variable number tandem repeat analysis of Mycobacterium bovis isolates from cattle in Brazil.

We performed spoligotyping and 12-mycobacterial interspersed repetitive unit-variable number tandem repeats (MIRU-VNTRs) typing to characterise Mycobacterium bovis isolates collected from tissue samples of bovines with lesions suggestive for tuberculosis during slaughter inspection procedures in abattoirs in Brazil. High-quality genotypes were obtained with both procedures for 61 isolates that were obtained from 185 bovine tissue samples and all of these isolates were identified as M. bovis by conventional identification procedures. On the basis of the spoligotyping, 53 isolates were grouped into nine clusters and the remaining eight isolates were unique types, resulting in 17 spoligotypes. The majority of the Brazilian M. bovis isolates displayed spoligotype patterns that have been previously observed in strains isolated from cattle in other countries. MIRU-VNTR typing produced 16 distinct genotypes, with 53 isolates forming eight of the groups, and individual isolates with unique VNTR profiles forming the remaining eight groups. The allelic diversity of each VNTR locus was calculated and only two of the 12-MIRU-VNTR loci presented scores with either a moderate (0.4, MIRU16) or high (0.6, MIRU26) discriminatory index (h). Both typing methods produced similar discriminatory indexes (spoligotyping h = 0.85; MIRU-VNTR h = 0.86) and the combination of the two methods increased the h value to 0.94, resulting in 29 distinct patterns. These results confirm that spoligotyping and VNTR analysis are valuable tools for studying the molecular epidemiology of M. bovis infections in Brazil.

Production and characterization of Clostridium perfringens recombinant ß toxoid.

In this work, we produced and evaluated a vaccine based on a ß toxoid of Clostridium perfringens type C produced in Escherichia coli (rBT). The non-toxic rBT was innocuous for mice and induced 14 IU mL(-1) of ß antitoxin in rabbits, complying with the European Pharmacopeia and CFR9 - USDA guidelines.

[Presence of IgM antibodies for Leptospira interrogans in wild animals from Tocantins State, 2002]. / Presença de anticorpos da classe IgM de Leptospira interrogans em animais silvestres do Estado do Tocantins, 2002.

Four hundred and twenty-seven serum samples of wild animals were tested against 18 serovars of Leptospira interrogans. Of 286 samples of Cebus apella, 46 (16.1%) were positive for the serovars pomona, brasiliensis, mini, swajizak, grippotyphosa, sarmin, fluminense, autumnalis, hebdomadis, guaratuba, javanica and icterohaemorrhagiae. Of 82 samples of Alouatta caraya, 2 (2.4%) were positive for the serovars mangus and fluminense. Of 31 samples of Nasua nasua, 4 (12.9%) were positive for the serovars fluminense and javanica, and of 10 samples of Cerdocyon thous, 2 (20 %) were positive for the serovars fluminense and brasiliensis. Seven samples of Dasyprocta sp, 6 of Tamandua tetradactyla and 5 of Euphractus sexcintus did not present reactivity.

Detection of toxins A/B and isolation of Clostridium difficile and Clostridium perfringens from dogs in Minas Gerais, Brazil

The objective of this study was to detect C. difficileA/B toxins and to isolate strains of C. perfringensand C. difficile from diarrheic and non-diarrheic dogs in Brazil. Stool samples were collected from 57 dogs, 35 of which were apparently healthy, and 22 of which were diarrheic. C. difficileA/B toxins were detected by ELISA, and C. perfringensand C. difficilewere identified by multiplex PCR. C. difficileA/B toxins were detected in 21 samples (36.8%). Of these, 16 (76.2%) were from diarrheic dogs, and five (23.8%) were from non-diarrheic dogs. Twelve C. difficile strains (21.1%) were isolated, of which ten were A+B+and two were A-B-. All non-toxigenic strains were isolated from non-diarrheic animals. The binary toxin gene cdtBwas found in one strain, which was A+B+and was derived from a non-diarrheic dog. C. perfringensstrains were isolated from 40 samples (70.2%). Of these, 18 (45%) were from the diarrheic group, and 22 (55%) belonged to the non-diarrheic group. All isolates were classified as C. perfringenstype A and there was an association between the detection of the cpegene and the presence of diarrhea. Interestingly, ten strains (25%) were positive for the presence of the cpb2gene. The high rate of detection of the A/B toxins in non-diarrheic dogs suggests the occurrence of subclinical disease in dogs or carriage of its toxins without disease. More studies are needed to elucidate the epidemiology of C. difficileand C. perfringensin dogs and to better our understanding of C. difficileas a zoonotic agent. This is the first study to report the binary toxin gene in C. difficilestrains isolated from dogs in Brazil.

Spoligotyping and variable number tandem repeat analysis of Mycobacterium bovis isolates from cattle in Brazil

We performed spoligotyping and 12-mycobacterial interspersed repetitive unit-variable number tandem repeats (MIRU-VNTRs) typing to characterise Mycobacterium bovis isolates collected from tissue samples of bovines with lesions suggestive for tuberculosis during slaughter inspection procedures in abattoirs in Brazil. High-quality genotypes were obtained with both procedures for 61 isolates that were obtained from 185 bovine tissue samples and all of these isolates were identified as M. bovis by conventional identification procedures. On the basis of the spoligotyping, 53 isolates were grouped into nine clusters and the remaining eight isolates were unique types, resulting in 17 spoligotypes. The majority of the Brazilian M. bovis isolates displayed spoligotype patterns that have been previously observed in strains isolated from cattle in other countries. MIRU-VNTR typing produced 16 distinct genotypes, with 53 isolates forming eight of the groups, and individual isolates with unique VNTR profiles forming the remaining eight groups. The allelic diversity of each VNTR locus was calculated and only two of the 12-MIRU-VNTR loci presented scores with either a moderate (0.4, MIRU16) or high (0.6, MIRU26) discriminatory index (h). Both typing methods produced similar discriminatory indexes (spoligotyping h = 0.85; MIRU-VNTR h = 0.86) and the combination of the two methods increased the h value to 0.94, resulting in 29 distinct patterns. These results confirm that spoligotyping and VNTR analysis are valuable tools for studying the molecular epidemiology of M. bovis infections in Brazil.

Detection of enterotoxin A and cytotoxin B, and isolation of Clostridium difficile in piglets in Minas Gerais, Brazil / Detecção da enterotoxina A e citotoxina B e isolamento de Clostridium difficile em leitões em Minas Gerais, Brasil

Clostridium difficile has emerged as a major cause of neonatal colitis in piglets, displacing classic bacterial pathogens. However, there is no information regarding the distribution of this microorganism in pig farms in Brazil. In the present study, the presence of toxins A/B and of C. difficile strains in stool samples from 60 diarrheic or non-diarrheic newborn piglets (one to seven days old), from 15 different farms, was studied. The presence of toxins A/B was detected by ELISA and PCR was used to identify toxin A, toxin B and binary toxin gene in each isolated strain. C. difficile A/B toxins were detected in ten samples (16.7 percent). Of these, seven were from diarrheic and three were from non-diarrheic piglets. C. difficile was recovered from 12 out of 60 (20 percent) fecal samples. Of those, three strains were non-toxigenic (A-B-) and nine were toxigenic. Of the nine toxigenic strains, four were A+B+ strains and five were A-B+ strains. The presence of binary toxin observed in the present study was much higher (50 percent) than in previously reported studies. All three non-toxigenic strains were isolated from otherwise healthy piglets. The results suggest the occurrence of neonatal diarrhea by C. difficile in farms in Brazil.

Clostridium difficile tem sido relatado como o principal causador de colite neonatal em suínos. Apesar da crescente importância deste agente, não há dados sobre infecções causadas por C. difficile em suínos no Brasil. O objetivo do presente estudo foi detectar as toxinas A/B e isolar C. difficile a partir de 60 amostras de fezes de leitões diarreicos ou apararentemente saudáveis, com no máximo sete dias de vida, e oriundos de 15 granjas diferentes. As toxinas A/B foram detectadas por ELISA e uma PCR multiplex foi utilizada para detecção dos genes responsáveis pela codificação das toxinas A, B e toxina binária. As toxinas A/B de C. difficile foram detectadas em dez amostras de fezes (16.7 por cento). Dessas, sete eram de animais diarreicos e três de leitões aparentemente saudáveis. Foi possível isolar C. difficile em 12 das 60 (20 por cento) amostras trabalhadas. Dessas, três estirpes eram não-toxigênicas (A-B-) e nove eram toxigênicas, sendo quatro caracterizadas como A+B+ e cinco como A-B+. O gene responsável pela codificação da toxina binária foi encontrado em 50 por cento das estirpes isoladas, proporção superior ao relatado em estudos anteriores. Todas as estirpes não toxigênicas foram isoladas de animais não diarreicos. Os resultados encontrados sugerem a ocorrência de diarreia por C. difficile em granjas no Brasil.

Padronização da titulação da toxina épsilon de Clostridium perfringens tipo D em linhagem contínua de células como alternativa ao bioensaio animal / Standardization of the titration of the epsilon toxin of Clostridium perfringens type D in cell line as an alternative to animal bioassay

Enterotoxemia, também chamada de doença do rim pulposo, doença que acomete os ruminantes domésticos, é causada pela ação da toxina épsilon produzida pelo Clostridium perfringens tipo D, um anaeróbio comumente isolado do solo e das fezes de animais sadios. O método tradicional de diagnóstico baseia-se na detecção e classificação dessa exotoxina no conteúdo intestinal por meio da soroneutralização em camundongos. Com isso, o objetivo deste estudo foi padronizar um teste para detecção e titulação dessa toxina in vitro e compará-lo ao fenômeno in vivo. Para isso, uma partida de toxina épsilon de Clostridium perfringens tipo D foi titulada em camundongos e em várias linhagens contínuas de células. Após a determinação da linhagem celular mais sensível, realizaram-se ensaios de titulação in vitro de diluições de uma partida de toxina, comparando-os com os títulos in vivo conhecidos. Os resultados foram agrupados, e foi desenvolvida a equação matemática que melhor adaptou-se aos intervalos trabalhados. A linhagem MDCK, além de mais sensível, demonstrou que o fenômeno observado in vitro pode ser expresso por meio da equação matemática que apresenta uma correlação de 98,33 por cento, com a dose mínima mortal determinada in vivo. Portanto, a linhagem MDCK permite titular a toxina épsilon de C. perfringens tipo D de forma específica e sensível, além de ser uma técnica prática, rápida e que dispensa o uso de animais.

Enterotoxemia (also called pulpy kidney disease) is an enteric disease, that affect ruminants, produced by epsilon toxin from Clostridium perfringens type D, an anaerobic commonly isolated from soil and feces of healthy animals. The diagnostic is based on detection of this exotoxin in the intestinal content by soroneutralization in mice. Therefore, this study aimed to standardize a test for detection and titration of the toxin in vitro, and compare it with the phenomenon in vivo. A volume of epsilon toxin was titrated in mice and in some cell lines. After concluding the most sensitive cell line, were held in vitro titrations of dilutions from a toxin wich one had in vivo titer known. The results were grouped and a mathematical equation was developed. MDCK cell line showed that the phenomenon observed in vitro can be expressed by a mathematical equation wich shows a correlation of 98.33 percent with a minimum lethal dose determined in vivo. Therefore, the soroneutralization using MDCK allows a specific, sensitive, practical, fast, and doesn't requere use of animal titration of epsilon toxin.

Botulismo tipo C em perus em Minas Gerais, Brasil / Type C botulism in turkeys in Minas Gerais, Brazil

O botulismo é uma intoxicação causada pela ingestão das toxinas produzidas pelo Clostridium botulinum, que acomete mamíferos e aves e é caracterizado por um quadro de paralisia flácida. Neste trabalho, é descrito um surto de botulismo em perus, ocorrido no município de Santa Luzia, região metropolitana de Belo Horizonte, Minas Gerais. Os animais apresentavam incoordenação motora, paralisia flácida das patas, asas e pescoço. Em um intervalo de 24 horas, todos os 29 animais do plantel vieram a óbito. Na necropsia, observou-se a presença de larvas de mosca no inglúvio. Nos soros coletados, foi identificada a toxina botulínica tipo C pelo teste de soroneutralização em camundongos.

Botulism is an intoxication caused by the ingestion of toxins produced by Clostridium botulinum. It affects mammals and birds, and is characterized by flaccid paralysis of the limbs. This report describes an outbreak of botulism in turkeys of various ages in the city of Santa Luzia, state of Minas Gerais, Brazil. The animals showed incoordination followed by flaccid paralysis involving the muscles of the legs, wings and neck. Within 24 hours, all 29 (100 percent) turkeys died. The post-mortem examination revealed the presence of fly larvae in the crop and the C. botulinum type C toxin was demonstrated in the sera of two affected animals by serum neutralization test.

Produção e caracterização de anticorpos monoclonais contra toxina épsilon de Clostridium perfringens Tipo D / Production and characterization of monoclonal antibodies against Clostridium perfringens Type D epsilon toxin

Clostridium perfringens tipo D é o agente etiológico da enterotoxemia em ruminantes, causada pela toxina épsilon e caracterizada por edema cardíaco, pulmonar, renal e cerebral. Anticorpos monoclonais contra toxina épsilon de C. perfringens tipo D foram produzidos a partir da fusão da linhagen de mieloma P3-X63-Ag8 653 com células do baço de camundongos Balb/c imunizados com o toxóide épsilon. Seis linhagens de híbridos secretores de anticorpos monoclonais das classes e IgM e IgG foram estabelecidas.

Clostridium perfringens type D is the aetiological agent of enterotoxemia in ruminants. The disease is caused by epsilon toxin characterized by cardiac, pulmonary, kidney and brain edema. Monoclonal antibodies were produced by using myeloma cell line P3-X63-Ag8 653 fused with spleen cells from Balb/c mice, immunized with epsilon toxoid of C. perfringens type D. Six hybrids were established secreting monoclonal antibodies of the IgM class and IgG3 subclass.

Genotipicação de Clostridium perfringens isolados de frangos de corte através da PCR múltipla / Genotyping Clostridium perfringens broiler chickens isolates by multiplex PCR products analyses

Clostridium perfringens (Cp) é uma bactéria aneróbica gram positiva que, além de provocar gangrena gasosa e enterotoxemia em humanos e animais, constitui-se na principal causa de enterite necrótica em aves de criações intensificadas. A identificação dos isolados foi realizada pela reação de lecitinase em ágar TSC-gema de ovo, colônias com dupla hemólise em ágar sangue desfibrinado de eqüino, coloração de Gram e provas bioquímicas. Das amostras analisadas, 171Cp foram isolados em jejuno e íleo de frangos de corte provenientes de um frigorífico da região de Pará de Minas-MG. Cp foi isolado em 62 (49,6 por cento) de 125 amostras de conteúdo lumenal de jejunos e em 109 (87,2 por cento) de igual número de íleos dos frangos de corte. Utilizando-se a técnica da PCR múltipla para genotipicacão das estirpes de Cp, de acordo com os genes para as toxinas principais e letais (cpa, cpb, etx e iA), da toxina cpb2 (cpb2) e enterotoxina (cpe), as estirpes de Cp isoladas foram classificadas em cinco tipos toxigênicos (A-E). Das 62 estirpes de Cp isoladas do jejuno, foram obtidos 42/62 (67,7 por cento) tipo A, 1/62 (1,6 por cento) tipo A com produto de amplificação para o gene da toxina beta2, 0/62 (0 por cento) tipo B, 17/62 (27,4 por cento) tipo C, 1/62 (1,6 por cento) tipo D. Das 109 amostras de Cp isolados do íleo das aves foram obtidos 62/109 (56,9 por cento) tipo A, 3/109 (2,7 por cento) tipo A com produto de amplificação para o gene da toxina beta2, 1/62 (0,9 por cento) tipo B, 38/109 (34,9 por cento) tipo C, 1/109 (0,9 por cento) tipo D. Cp A (60,8 por cento) e Cp C (32,2 por cento) foram os tipos toxigênicos predominantes em conteúdo intestinal de frango de corte. Cinco (2,9 por cento) das 171 amostras de Cp isolados não foram tipificadas. Não foram detectados os genes codificadores das toxinas iota (iA) e enterotoxina (cpe) em nenhuma das 171 estirpes de Cp caracterizados.

Clostridium perfringens (Cp) is an anaerobic gram-positive bacterium which causes gaseous gangrene and enterotoxaemias in humans and domestic animals, besides being the primary cause of necrotic enteritis in poultry. Cp isolates were preliminary identified according to the lecithinase test on agar TSC-egg yolk, colony with double haemolysis in desfibrinated horse blood agar, Gram staining and biochemical tests. Cp isolates (171) were obtained from the intestinal content of broiler chickens sampled in a slaughterhouse in Pará de Minas city, MG, Brazil. Cp was isolated in 62/125 (49.6 percent) strains from jejunum content and in 109/125 (87.2 percent) of ileum. Cp strains were classified into five toxigenic types (A-E), using multiplex PCR assay for genotyping of the principal and lethal toxins in the detection of genes coding for toxins alfa, beta, epsilon e iota, nomely genes cpa, cpb, etx e iA genes, beta2 toxin (cpb2) and enterotoxin (cpe). From a total of 62 Cp jejunum isolates obtained 42/62 (67.7 percent) were type A, 1/62 (1.6 percent) type A with the amplification of products for beta2 toxin gene (A/B2), 0/62 (0 percent) type B, 17/62 (27.4 percent) type C and 1/62 (1.6 percent) type D. A total of 109 ileum Cp isolates were obtained being 62/109 (56.9 percent) type A, 3/109 (2.7 percent) type A/B2 toxin gene, 1/62 (0.9 percent) type B, 38/109 (34.9 percent) type C, 1/109 (0.9 percent) type D. Cp A (60.8 percent) and Cp C (32.2 percent) toxigenic types were the most prevalent types in the analyzed intestinal contents of broiler chickens Cp A 104/171 (60.8 percent) and 55/171 (32.2 percent) toxigenic types which were the most prevalent types analyzed into two partes of the intestinal content of broiler chickens. Five (2.9 percent) out of 171 Cp isolates were not typified. The iota toxin (iA) and enterotoxin gene (cpe) codifying genes were not identified.

Concentração inibitória mínima (CIM) de oito antimicrobianos frente isolados de Streptococcus suis / Mininum inhibitory concentration of eigth antibiotics toward isolates of Streptococcus suis

Avaliou-se a concentração inibitória mínima (CIM) de 75 isolados de Streptococcus suis frente a oito antimicrobianos freqüentemente utilizados no controle da infecção por esse microrganismo. A CIM foi realizada em placas previamente preparadas com ágar sangue, contendo concentrações variando de 0,25 a 256 μg/ml dos seguintes antimicrobianos: amoxicilina, ampicilina, penicilina, ceftiofur sódico, florfenicol, lincomicina, penicilina, sulfametoxazol-trimetoprim e tetraciclina. A amoxicilina nas concentrações de 1 e 2 μg/ml, o florfenicol a 1 μg/ml e o sulfametoxazol-trimetoprim nas concentrações de 2 e 8 μg/ml, foram os antimicrobianos frente aos quais os microrganismos apresentaram menor resistência. Em contraste, a ampicilina, tetraciclina, ceftiofur sódico e lincomicina, foram menos efetivos, apresentando CIM de (64 e 128 μg/ml), (64 e 128 μg/ml), (128 e 256 μg/ml) e (>256 μg/ml), respectivamente. Os resultados deste estudo demonstram que a amoxicilina e o florfenicol são os antibióticos de escolha para o tratamento de infecções pelo S. suis em suínos.

The minimun inhibitory concentration (MIC) was determined toward amoxicilin, ampicilin, penicillin, ceftiofur, florfenicol, lincomycin, trimethoprim-sulfadiazine and tetracycline for 75 strains of Streptococcus suis. The MIC was performed on sheep blood agar plates containing concentrations varying from 0,25 to 256 μg/ml of the antibiotic described above. The amoxicilin in the concentrations of 1 and 2 μg/ml, florfenicol in the concentration of 1 μg/ml and trimethoprim-sulfadiazine in the concentrations of 2 e 8 μg/ml, were the antibiotics that presented minor resistance. In contrast, the ampicilin, tetracycline, ceftiofur and lincomycin, presented MIC of 64 and 128 μg/ml, 64 and 128 μg/ml, 128 and 256 μg/ml and >256 μg/ml, respectively. The results of this study show that the amoxicilin and florfenicol are the antibiotics of choice for the treatment of diseases by S. suis in swine.

Botulismo em ruminantes causado pela ingestão de cama-de-frango / Botulism in ruminants being fed with poultry litter

Botulismo é uma intoxicação causada pela ingestão das toxinas produzidas pelo Clostridium botulinum, que acomete mamíferos e aves. Neste trabalho é descrito um surto de botulismo em ruminantes, ocorrido em duas propriedades localizadas no município de Patos, no Estado da Paraíba, Brasil. Em uma das propriedades, de um total de 88 bovinos, 85 (96,6 por cento) vieram a óbito. Na segunda, morreram 145 ovinos (96,7 por cento), 233 caprinos (57,8 por cento) e 30 bovinos (96,8 por cento). Os animais acometidos apresentavam paralisia progressiva, dificuldade de locomoção, sialorréia e dispnéia. A morte ocorreu entre 24 e 48 horas após o início dos sinais, por parada cardio-respiratória. Nenhuma alteração significativa foi observada no exame post-mortem. O diagnóstico de botulismo foi confirmado pela demonstração das toxinas C e D no conteúdo intestinal e na cama-de-frango utilizada na alimentação dos animais, pela técnica de soroneutralização em camundongos.

Botulism is a poisoning caused by the ingestion of toxins produced by Clostridium botulinum, that infects mammals and birds. This article reports an outbreak of botulism in two different flocks of ruminants at Paraíba, Brazil. In one, 85 out of 88 (96.6 percent) cattles died. In the other, 145 (96.7 percent) sheeps, 233 (57.8 percent) goats and 30 (96.8 percent) cattles died. Clinical signs were progressive paralysis, difficulties in moving, sialorrhoe and dyspnoe. Death occurred 24 to 48 hours after the beginning of clinic signs and at post-mortem examination no noteable changes were observed. Type C and D toxins were demonstrated in the intestinal contents and poultry litter by neutralization test in mice.

Botulismo tipo C em ganso ocorrido em Minas Gerais, Brasil / Type C botulism in a goose at Minas Gerais, Brazil

Botulismo é uma intoxicação causada pela ingestão das toxinas produzidas pelo Clostridium botulinum, que acomete mamíferos e aves, caracterizando-se por um quadro de paralisia flácida. Neste trabalho, é descrito um caso de botulismo em ganso, ocorrido no município de Santa Luzia, região metropolitana de Belo Horizonte, no Estado de Minas Gerais. Ao exame clínico, o animal apresentava-se com um quadro de paralisia flácida dos músculos do pescoço, das pernas e asas, além de apresentar ainda desprendimento de penas. A necropsia não revelou lesões significativas. Foi colhido o soro do animal e submetido ao teste de soroneutralização em camundongo, que identificou a toxina de C. botulinum tipo C.

Botulism is an intoxication caused by the ingestion of toxins produced by Clostridium botulinum, that affects mammals and birds, characterized by a flaceid paralysis. This report describes a case of botulism in a goose in Santa Luzia, Minas Gerais State, Brazil. Clinical examinations showed dropping feathers and flaccid paralysis involving the muscles of the wings, legs and neck. post-mortem examination showed no significant gross or macroscopic lesions C. botulinum type C toxin was demonstrated in the serum of the affected animal through serum neutralization test in mice.

Primeiro relato no Brasil de mastite necrótica bovina por Clostridium perfringens tipo A / First report in Brazil of bovine necrotic mastitis due to Clostridium perfringens type A

Relata-se o primeiro caso no Brasil de mastite bovina por Clostridium perfringens tipo A. O quadro clínico caracterizou-se por necrose da papila mamária e porção ventral do quarto afetado. O agente foi isolado em cultura pura e identificado como tipo A por PCR a partir do leite do quarto mamário afetado.