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Comparison of a new, bismuth-iron-sulfite-cycloserine agar for isolation of Clostridium perfringens with the tryptose-sulfite-cycloserine and blood agars.

A new differential and selective, bismuth-iron-sulfite-cycloserine (BISC) medium, for isolation and enumeration of Clostridium perfringens from food and feces, was developed. The medium was compared with the widely-used tryptose-sulfite-cycloserine (TSC) medium and blood agar (BA) in recovering actively growing cells, cold- (refrigerated and frozen) stressed, and heat-stressed C. perfringens cells, and heat-activated spores from human feces. Both selective media were satisfactory in recovering actively growing cells and heat-activated spores of C. perfringens. Both were inferior to non-inhibitory blood agar in recovering heat or cold-stressed cells. The advantages of the new BISC medium over the TSC medium were: elimination of the need to prepare pour- or overlay-agar plates, which simplified inoculation of specimens on the medium and simplified the subcultures of colonies for confirmatory identification. All colonies of C. perfringens developed on BISC were black or dark gray. This was contrary to TSC medium, which gave, on average, 39.6% of white colonies when inoculated with the pure cultures of C. perfringens.

Bacteriological Techniques↗

Molecular analysis of the virulence determinants of Clostridium perfringens associated with foal diarrhoea.

During an epidemiological study of foal diarrhoea, over half of the cases yielded Clostridium perfringens which was significantly associated with disease (Netherwood et al., 1996b). However, the association could not be accounted for by enterotoxigenic isolates which had a low prevalence (Netherwood et al., 1997). Nonetheless, we have hypothesized that the association may be caused by a pathogenic sub-population which would be significantly more common amongst C. perfringens-positive cases compared with C. perfringens-positive healthy controls if it acted as a pathogen when present. Conversely, if foal diarrhoea caused by C. perfringens was dependent on a predisposing factor, then such an association might not be evident. As a first step to determine if a molecular marker was more frequently to be found in C. perfringens-positive cases than controls, we have genotyped the study isolates (up to five per foal) by polymerase chain reaction (PCR) based on the published gene sequences for the major lethal toxins alpha, beta, epsilon and iota as well as for theta toxin, large and small sialidases, hyaluronidase and virulence regulation. Isolates of major toxin types B, C, D and E, or isolates which were untypeable, were isolated from less than 15% of C. perfringens-positive foals and these were not associated with diarrhoea nor were they more commonly found in C. perfringens-positive cases. Isolates of type A were found in more than 90% of all C. perfringens-positive foals. A number of different genotypes were identified by their different patterns of gene possession but types without any of the genes for theta toxin, large and small sialidases, hyaluronidase and virulence regulation were found in only 10% of positive foals. Only type A isolates with all of these genes were associated with diarrhoea overall but they were not more commonly isolated from C. perfringens-positive cases than controls. In conclusion, genotyping by the sequenced virulence genes did not identify a marker for a sub-population of C. perfringens which may be acting more frequently as a pathogen when present.

Amino Acid Sequence↗

A fatal sandwich.

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Bacterial Toxins↗

Application of serological typing to the investigation of outbreaks of Clostridium perfringens food poisoning, 1970-1978.

Serological typing was used as an epidemiological tool in the investigation of 524 outbreaks of Clostridium perfringens food poisoning in the United Kingdom and 37 outbreaks in other countries. Five thousand five hundred and fifty-four (77%) of 7245 strains of C. perfringens associated with the 561 outbreaks were typable with the 75 Food Hygiene Laboratory antisera; in 354 (63%) of these outbreaks a specific serotype was established as being responsible for the outbreak. An assessment is made of the ability of two additional sets of antisera, prepared against 34 American and 34 Japanese strains of C. perfringens, to increase the number of strains which can be typed. The extent of cross-reaction between the three sets of antisera was determined and the results are discussed in relation to the source and history of the type strains.

Clostridium Infections↗

Large outbreaks of Clostridium perfringens food poisoning associated with the consumption of boiled salmon.

Five large outbreaks of food poisoning are described in which clinical, epidemiological or laboratory data indicated Clostridium perfringens as the causative organism. The foodstuff common to all incidents was boiled salmon served cold as an hors d 'oeuvre. In all cases the fish had been subject to a long period of cooling or storage between boiling and consumption. It is thought that multiplication of the organism occurred during this time. Recommendations are made for the avoidance of further similar incidents.

Animals↗

Techniques known to prevent post-operative wound infection.

The most important factors in controlling postsurgical sepsis are appropriate surgical judgment and technique. Efficacious prophylactic antibiotics, when indicated, also significantly reduce the postoperative infection rate. Among the other techniques often heralded as important adjunctive measures, only duration of preoperative hospitalization, preoperative bathing, use of electrocautery, preoperative hair removal, use of prophylactic drains, and duration of operation are of proven significance.

Anti-Bacterial Agents↗

Wound infection: a review of diagnosis and treatment.

Surgical wounds can become infected by a variety of organisms, leading to a variety of clinical courses. In this article, several types of infections, therapies, prognoses and prophylaxes are discussed. Skin and subcutaneous tissues infect with a range of gram-positive and gram-negative aerobic and anerobic bacteria. Gangrenous infections, whether necrotizing fasciitis or bacterial synergistic gangrene, can be devastating or fatal. Gangrenous infection in the muscle is also discussed, and various treatments outlined. The intention is to familiarize readers with the predisposing conditions, clinical manifestations and therapeutic alternatives for these surgical infections.

Bacterial Infections↗

Occurrence of Clostridium perfringens beta2-toxin amongst animals, determined using genotyping and subtyping PCR assays.

Clostridium perfringens isolates are currently classified into one of five biotypes on the basis of the differential production of alpha-, beta-, epsilon- and iota-toxins. Different biotypes are associated with different diseases of man and animals. In this study a multiple PCR assay was developed to detect the genes encoding these toxins. In addition, detection of the genes encoding the C. perfringens enterotoxin and beta2-toxin allowed subtyping of the bacteria. C. perfringens isolates taken from a variety of animals, including foals, piglets or lambs, were genotyped using this assay. Most of the isolates were found to be genotype A and the gene encoding beta2-toxin [corrected] was present in 50% of the isolates genotyped. A significant association between C. perfringens possessing the beta2-toxin gene and diarrhoea in piglets was identified, suggesting that beta2-toxin may play a key role in the pathogenesis of the disease.

Animals↗

Antibody response in goats vaccinated with liposome-adjuvanted Clostridium perfringens type D epsilon toxoid.

A trial was performed using 20 goats to evaluate the antibody responses to a liposome-adjuvanted Clostridium perfringens epsilon toxoid vaccine (LIPV). The antibody response was compared with those produced by epsilon toxoid vaccines prepared using aluminium hydroxide (ALV) and incomplete Freud's adjuvant (FAV). The animals were allocated to four groups at the beginning of the trial. The animals in group 1 were vaccinated with ALV, while the animals in group 2 received FAV and those in groups 3 and 4 were vaccinated with LIPV. The animals in groups 1 to 3 received three doses of the corresponding vaccine at intervals of three weeks, while those in group 4 received only 1 dose of vaccine at the beginning of the trial. A blood sample was obtained from all the goats at the beginning of the trial and then weekly for 8 weeks. The samples were analysed for epsilon toxoid antibodies by an indirect ELISA technique. No major clinical abnormalities were observed in the animals after vaccination, with the exception of those that received the FAV, which experienced transient lameness. The highest antibody response was observed in the animals vaccinated with FAV, but they presented moderate to severe inflammatory tissue reactions at the injection site. Moderately high antibody responses were obtained with the ALV, with which only minor local reactions were observed. No significant antibody responses were obtained with the LIPV, nor were local reactions observed.

Aluminum Hydroxide↗

[The necrotizing enteritis by Clostridium perfringens type C in piglets: II. Molecular epidemiology study].

Investigations were performed on shedding of C. perfringens in sows from four different pig farms. In two farms where no outbreaks of necrotizing enteritis had been observed, no strains of C. perfringens producing beta-toxin were detected in the faeces of sows. In contrast, C. perfringens strains producing beta-toxin were detected in sows on both farms suffering outbreaks of acute necrotizing enteritis. Strains of C. perfringens producing beta-toxin were invariably positive for the beta 2-toxin gene. However, strains carrying the beta 2-toxin gene only (i.e. negative for beta-toxin) were present in animals on all farms with roughly similar frequencies (mean 28.2% carriers). Some sows carried C. perfringens strains of both toxin genotypes simultaneously. Whereas these data further support the role of betatoxin as a cause of necrotizing enteritis, the role of beta 2-toxin in intestinal disease of piglets remains unclear. To establish the role of faecal shedding vs. environmental contamination as reservoirs of C. perfringens type C, strains were isolated from teats and feedlot trough swabs (toxin genotype beta/beta 2), as well as from fodder (genotype beta 2). However, sows carried this pathogen intermittently and in small numbers. This renders an individual, reliable diagnosis of carrier sows very difficult. Ribotyping of 34 C. perfringens isolates of different toxin genotypes showed five distinct profiles. Different toxin genotypes can belong to the same ribotype, and the same toxin genotype can be present in different ribotypes. Thus, even if a majority (79.4%) of strains investigated in a limited geographic region belonged to ribotype 1, ribotyping offered discrimination of strains beyond toxin typing.

Animals↗

[Tetanus in cats: 3 case descriptions].

Three cats with spasticity on one leg or on all four limbs were presented between 1996 and 1998 at the Department of clinical veterinary medicine, Section of neurology, Vetsuisse-Faculty of Bern. The presumptive diagnosis was tetanus. A focal form was present in two cases and generalised tetanus in one cat. All cats had a history of injury at the affected legs respectively at the neck. The first clinical signs were seen between two days and three weeks after injury. The bacteriologic examination of serous fluid from the site of injury revealed an infection with Clostridium. EMG in one cat during anaesthesia showed motor united potentials (MUPs) on the spastic leg. All patients received antibiotics (Penicillin, respectively Amoxicillin/Clavulanic acid and Metronidazol). Supportive aid were initially sedation, wound revision and in one cat nutrition through oesophageal sonde. In a second phase physiotherapy was performed. All three animals were significantly better after a couple of weeks, two cats were without symptoms after eight and five weeks respectively.

Animals↗