scholarly journals Detection of Enterotoxigenic Clostridium perfringens Type A Isolates in American Retail Foods

2004 ◽  
Vol 70 (5) ◽  
pp. 2685-2691 ◽  
Author(s):  
Qiyi Wen ◽  
Bruce A. McClane

ABSTRACT Currently there is only limited understanding of the reservoirs for Clostridium perfringens type A food poisoning. A recent survey (Y.-T. Lin and R. Labbe, Appl. Environ. Microbiol. 69:1642-1646, 2003) of non-outbreak American retail foods did not identify the presence of a single C. perfringens isolate carrying the enterotoxin gene (cpe) necessary for causing food poisoning. The present study revisited this issue, using revised methodology and food sampling strategies. In our survey, cpe-positive C. perfringens isolates were detected in ∼1.4% of ∼900 surveyed non-outbreak American retail foods. Interestingly, those enterotoxigenic isolates in non-outbreak foods appear indistinguishable from C. perfringens isolates known to cause food poisoning outbreaks: i.e., the enterotoxigenic retail food isolates all carry a chromosomal cpe gene, are classified as type A, and exhibit exceptional heat resistance. Collectively, these findings indicate that some American foods are contaminated, at the time of retail purchase, with C. perfringens isolates having full potential to cause food poisoning. Furthermore, demonstrating that type A isolates carrying a chromosomal cpe gene are the enterotoxigenic isolates most commonly present in foods helps to explain why these isolates (rather than type A isolates carrying a plasmid cpe gene or cpe-positive type C or D isolates) are strongly associated with food poisoning outbreaks. Finally, since type A chromosomal cpe isolates present in the surveyed raw foods exhibited strong heat resistance, it appears that exceptional heat resistance is not a survivor trait selected for by cooking but is instead an intrinsic trait possessed by many type A chromosomal cpe isolates.

2012 ◽  
Vol 80 (12) ◽  
pp. 4354-4363 ◽  
Author(s):  
Menglin Ma ◽  
Jihong Li ◽  
Bruce A. McClane

ABSTRACTClostridium perfringenstype C strains are the only non-type-A isolates that cause human disease. They are responsible for enteritis necroticans, which was termed Darmbrand when occurring in post-World War II Germany. Darmbrand strains were initially classified as type F because of their exceptional heat resistance but later identified as type C strains. Since only limited information exists regarding Darmbrand strains, this study genetically and phenotypically characterized seven 1940s era Darmbrand-associated strains. Results obtained indicated the following. (i) Five of these Darmbrand isolates belong to type C, carry beta-toxin (cpb) and enterotoxin (cpe) genes on large plasmids, and express both beta-toxin and enterotoxin. The other two isolates arecpe-negative type A. (ii) All seven isolates produce highly heat-resistant spores withD100values (the time that a culture must be kept at 100°C to reduce its viability by 90%) of 7 to 40 min. (iii) All of the isolates surveyed produce the same variant small acid-soluble protein 4 (Ssp4) made by type A food poisoning isolates with a chromosomalcpegene that also produce extremely heat-resistant spores. (iv) The Darmbrand isolates share a genetic background with type A chromosomal-cpe-bearing isolates. Finally, it was shown that both thecpeandcpbgenes can be mobilized in Darmbrand isolates. These results suggest thatC. perfringenstype A and C strains that cause human food-borne illness share a spore heat resistance mechanism that likely favors their survival in temperature-abused food. They also suggest possible evolutionary relationships between Darmbrand strains and type A strains carrying a chromosomalcpegene.


2007 ◽  
Vol 73 (22) ◽  
pp. 7218-7224 ◽  
Author(s):  
Jihong Li ◽  
Sameera Sayeed ◽  
Bruce A. McClane

ABSTRACT In the United States and Europe, food poisoning due to Clostridium perfringens type A is predominantly caused by C. perfringens isolates carrying a chromosomal enterotoxin gene (cpe). Neither the reservoir for these isolates nor the point in the food chain where these bacteria contaminate foods is currently understood. Therefore, the current study investigated whether type A isolates carrying a chromosomal cpe gene are present in two potential reservoirs, i.e., soil and home kitchen surfaces. No C. perfringens isolates were recovered from home kitchen surfaces, but most surveyed soil samples contained C. perfringens. The recovered soil isolates were predominantly type A, but some type C, D, and E soil isolates were also identified. All cpe-positive isolates recovered from soil were genotyped as type A, with their cpe genes on cpe plasmids rather than the chromosome. However, two cpe-positive soil isolates did not carry a classical cpe plasmid. Both of those atypical cpe-positive soil isolates were sporulation capable yet failed to produce C. perfringens enterotoxin, possibly because of differences in their upstream promoter regions. Collectively these results suggest that neither soil nor home kitchen surfaces represent major reservoirs for type A isolates with chromosomal cpe that cause food poisoning, although soil does appear to be a reservoir for cpe-positive isolates causing non-food-borne gastrointestinal diseases.


2008 ◽  
Vol 74 (11) ◽  
pp. 3328-3335 ◽  
Author(s):  
Benjamin Orsburn ◽  
Stephen B. Melville ◽  
David L. Popham

ABSTRACT The endospores formed by strains of type A Clostridium perfringens that produce the C. perfringens enterotoxin (CPE) are known to be more resistant to heat and cold than strains that do not produce this toxin. The high heat resistance of these spores allows them to survive the cooking process, leading to a large number of food-poisoning cases each year. The relative importance of factors contributing to the establishment of heat resistance in this species is currently unknown. The present study examines the spores formed by both CPE+ and CPE− strains for factors known to affect heat resistance in other species. We have found that the concentrations of DPA and metal ions, the size of the spore core, and the protoplast-to-sporoplast ratio are determining factors affecting heat resistance in these strains. While the overall thickness of the spore peptidoglycan was found to be consistent in all strains, the relative amounts of cortex and germ cell wall peptidoglycan also appear to play a role in the heat resistance of these strains.


2012 ◽  
Vol 42 (8) ◽  
pp. 1450-1456 ◽  
Author(s):  
Thais Sebastiana Porfida Ferreira ◽  
Andrea Micke Moreno ◽  
Renata Rodrigues de Almeida ◽  
Cleise Ribeiro Gomes ◽  
Debora Dirani Sena de Gobbi ◽  
...  

Clostridium perfringens is an anaerobic Gram-positive bacterium known as common pathogen for humans, for domestic and wildlife animals. Although infections caused by C. perfringens type C and A in swine are well studied, just a few reports describe the genetic relationship among strains in the epidemiological chain of swine clostridioses, as well as the presence of the microorganism in the slaughterhouses. The aim of the present study was to isolate C. perfringens from feces and carcasses from swine slaughterhouses, characterize the strains in relation to the presence of enterotoxin, alpha, beta, epsilon, iota and beta-2 toxins genes, using polymerase chain reaction (PCR) and comparing strains by means of Pulsed field gel electrophoresis (PFGE). Clostridium perfringens isolation frequencies in carcasses and finishing pig intestines were of 58.8% in both types of samples. According to the polymerase chain reaction assay, only alfa toxin was detected, being all isolates also negative to enterotoxin and beta2 toxin. Through PFGE technique, the strains were characterized in 35 pulsotypes. In only one pulsotype, the isolate from carcass sample was grouped with fecal isolate of the same animal, suggesting that the risk of cross-contamination was low. Despite the high prevalence of C. perfringens in swine carcasses from the slaughterhouses assessed, the risk of food poisoning to Brazilian pork consumers is low, since all strains were negative to cpe-gene, codifying enterotoxin.


2002 ◽  
Vol 70 (8) ◽  
pp. 4261-4272 ◽  
Author(s):  
Kazuaki Miyamoto ◽  
Ganes Chakrabarti ◽  
Yosiharu Morino ◽  
Bruce A. McClane

ABSTRACT Clostridium perfringens type A isolates causing food poisoning have a chromosomal enterotoxin gene (cpe), while C. perfringens type A isolates responsible for non-food-borne human gastrointestinal diseases carry a plasmid cpe gene. In the present study, the plasmid cpe locus of the type A non-food-borne-disease isolate F4969 was sequenced to design primers and probes for comparative PCR and Southern blot studies of the cpe locus in other type A isolates. Those analyses determined that the region upstream of the plasmid cpe gene is highly conserved among type A isolates carrying a cpe plasmid. The organization of the type A plasmid cpe locus was also found to be unique, as it contains IS1469 sequences located similarly to those in the chromosomal cpe locus but lacks the IS1470 sequences found upstream of IS1469 in the chromosomal cpe locus. Instead of those upstream IS1470 sequences, a partial open reading frame potentially encoding cytosine methylase (dcm) was identified upstream of IS1469 in the plasmid cpe locus of all type A isolates tested. Similar dcm sequences were also detected in several cpe-negative C. perfringens isolates carrying plasmids but not in type A isolates carrying a chromosomal cpe gene. Contrary to previous reports, sequences homologous to IS1470, rather than IS1151, were found downstream of the plasmid cpe gene in most type A isolates tested. Those IS1470-like sequences reside in about the same position but are oppositely oriented and defective relative to the IS1470 sequences found downstream of the chromosomal cpe gene. Collectively, these and previous results suggest that the cpe plasmid of many type A isolates originated from integration of a cpe-containing genetic element near the dcm sequences of a C. perfringens plasmid. The similarity of the plasmid cpe locus in many type A isolates is consistent with horizontal transfer of a common cpe plasmid among C. perfringens type A strains.


1988 ◽  
Vol 101 (3) ◽  
pp. 661-667 ◽  
Author(s):  
P. W. Collier ◽  
J. C. M. Sharp ◽  
Aida F. MacLeod ◽  
G. I. Forbes ◽  
F. Mackay

SUMMARYDuring the 10-year period 1978–87 there were 48 outbreaks of food poisoning in Scottish hospitals affeeting a total of 2287 persons of whom 12 died. This compared with 50 outbreaks during the previous 5 years (1973–77) when over 1500 persons and 7 deaths were recorded. Although the incidence of outbreaks has decreased the average number of persons affected in outbreaks has increased. A marked reduction was seen in the incidence of outbreaks due to Clostridium perfringens, in contrast to foodborne salmonellosis which remains a problem. Thirty-four hospitals, of which 10 reported two or more outbreaks, were involved. The type of hospitals most frequently affected were general (14), psychiatric (13), geriatric (9) and hospitals for the mentally subnormal (7). Meat, including poultry meat, was incriminated in over 90% of outbreaks where a food vehicle was identified. In modern or re-equipped kitchens cooking in advance with subsequent reheating is being progressively discontinued as more food is being cooked on the day of consumption, a practice which may readily explain the decreasing incidence of outbreaks due to Cl. perfringens. Bacterial cross-contamination from poultry-meat and other raw foods, compounded by inadequate temperature control, however, continues to be a problem in some hospitals. It is too early as yet to determine whether the removal of Crown immunity will have any effect on the future incidence of hospital ‘food poisoning’.


1970 ◽  
Vol 16 (5) ◽  
pp. 331-338 ◽  
Author(s):  
A. H. W. Hauschild ◽  
L. Niilo ◽  
W. J. Dorward

The possible involvement of the following factors in Clostridium perfringens type A enteritis was investigated: cellular constituents of C. perfringens, extracellular components, gas generated by growing cultures, and organic acid production. Lambs were used as experimental animals, and all materials were administered by the intraduodenal route.Vegetative cells (1.5 to 5 × 109) administered along with fresh medium caused diarrhea while 100 times the number of cells in spent medium or saline had no effect. Sporulating cells in saline, and their extracts, caused profuse diarrhea. Supernatant fluids from both vegetative and sporulating cultures concentrated by dialysis against polyethylene glycol had no effect, but diarrhea was produced by supernatant fluids from vegetative cultures concentrated by rotary evaporation, and by the gas generated during growth. The effect of the gas was simulated with hydrogen. The pH of the fluid in the small intestine remained constant during C. perfringens enteritis.It was concluded (i) that the factor responsible for experimental enteritis in lambs resulting from infection of the small intestine with vegetative cells of C. perfringens is produced in situ, and (ii) that of the various factors tested, a heat-sensitive, nondialyzable component of sporulating cells is most likely identical with the main enteropathogenic factor in C. perfringens enteritis.


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