Clostridium Perfringens Food Poisoning Research Articles

Clostridium perfringens food poisoning: An update on toxins, occurrence, diseases and detection techniques

Clostridium perfringens food poisoning: An update on toxins, occurrence, diseases and detection techniques

Dangers of Clostridium perfringens food poisoning in psychiatric patients.

Clostridium perfringens food poisoning can be fatal in patients with chronic constipation. We report the investigation and management of a probable outbreak of C. perfringens food poisoning among psychiatric patients in Cape Town, South Africa, in 2013.

Epidemiological studies on Clostridium perfringens food poisoning in retail foods.

Clostridium perfringens is an important anaerobic pathogen causing food-borne gastrointestinal (GI) diseases in humans and animals. Meat and meat products are the most common vehicles of C. perfringens type A food poisoning. Contamination of meat by the intestinal contents of slaughtered animals may serve as an important source of this pathogen to the food supply. One hundred and fifty-five non-outbreak food samples were obtained from meat and retail food and examined for the presence of C. perfringens. Multiplex polymerase chain reaction assay to determine the toxin genotype of C. perfringens isolates, and extraction and purification of C. perfringens enterotoxin from enterotoxin gene (cpe)-positive isolates were carried out. The homogeneity of the purified enterotoxin was demonstrated by polyacrylamide gel electrophoresis. In addition, stool samples were collected from 150 persons who had been in contact with animals, and enzyme-linked immunosorbent assays were carried out for the qualitative determination of C. perfringens enterotoxin in the stool samples. The results demonstrated that approximately 2.6% of the tested meat and retail meat samples were contaminated with cpe-positive C. perfringens. The recommended laboratory criteria used to implicate C. perfringens in food-borne disease should involve the detection of C. perfringens enterotoxin production or the presence of the cpe gene in foods or faeces, or in the suspected C. perfringens isolates. In the present study some isolates such as tuna contained the enterotoxin gene although they had a low count of C. perfringens.

Location and stoichiometry of the protease CspB and the cortex-lytic enzyme SleC in Clostridium perfringens spores

The protease CspB and the cortex-lytic enzyme SleC are essential for peptoglycan cortex hydrolysis during germination of spores of the Clostridium perfringens food poisoning isolate SM101. In this study, Western blot analyses were used to demonstrate that CspB and SleC are present exclusively in the C. perfringens SM101 spore coat layer fraction and absent in the lysate from decoated spores and from the purified inner spore membrane. These results indicate why decoating treatments greatly reduce both germination and apparent viability of C. perfringens spores in the absence of an exogenous lytic enzyme. In addition, quantitative Western blot analyses showed that there are approximately 2000 and 130,000 molecules of CspB and pro-SleC, respectively, per C. perfringens SM101 spore, consistent with CspB's role in acting catalytically on pro-SleC to convert this zymogen to the active enzyme.

A Mild Outbreak of Gastroenteritis in Long-Term Care Facility Residents Due toClostridium perfringens, Australia 2009

Clostridium perfringens food poisoning is a commonly cited cause of gastroenteritis outbreaks among elderly long-term care facility (LTCF) residents, yet little is known about the natural history of disease in this vulnerable group. In July 2009, an investigation into diarrheal illness among LTCF residents was commenced. An environmental health investigation and retrospective cohort study were undertaken to confirm the outbreak, to identify a source and mode of transmission, and to implement public health measures to prevent further cases. Menu listings and food safety program details were obtained and food-handling practices were observed. Clinical notes of all residents were reviewed. A possible case was defined as any resident developing one or more acute loose stool episodes between the evenings of 23 July and 27 July. Fifty-two residents (41%) had been ill with diarrhea, and eight residents had fecal samples positive for C. perfringens enterotoxin. LTCF staff failed to perform routine temperature checks on hot foods before the outbreak. A sweet-and-sour pork lunch served on 23 July was implicated in causing residents' illness, but no residual food remained for microbiological testing. Independent associations with illness were demonstrated among residents living in two wings of the facility that received a standard level of service, whereas an inverse association with illness was shown among residents living in an "extra service" wing. Male residents were also more likely to become ill. Illness was mild with case patients reporting a median of two loose stools (range 1-12). C. perfringens is an important cause of both foodborne and nonfoodborne gastroenteritis outbreaks in LTCF, but may be missed due to the often mild nature of illness. This investigation highlights the potential burden of C. perfringens disease among vulnerable LTCF populations. To prevent C. perfringens outbreaks, facilities must adhere to food safety plans and ensure high standards of infection control practice.

The protease CspB is essential for initiation of cortex hydrolysis and dipicolinic acid (DPA) release during germination of spores of Clostridium perfringens type A food poisoning isolates

The genome of the Clostridium perfringens food poisoning isolate SM101 encodes a subtilisin-like protease, CspB, upstream of the sleC gene encoding the enzyme essential for degradation of the peptidoglycan cortex during spore germination. SleC is an inactive pro-SleC in dormant spores that is converted to active SleC during spore germination and Csp proteases convert pro-SleC to the active enzyme in vitro. In this work, the germination and viability of spores of a cspB deletion mutant of strain SM101, as well as cspB expression, were studied. The cspB gene was expressed only during sporulation, and only in the mother cell compartment. cspB spores were unable to germinate significantly with either a rich nutrient medium, KCl, or a 1 : 1 chelate of Ca(2+) and dipicolinic acid (DPA); the viability of these spores was approximately 10(4)-fold lower than that of wild-type spores, although cspB and wild-type spores had similar viability on plates containing lysozyme, and cspB spores could not process inactive pro-SleC into active SleC during spore germination. Germination of cspB spores was blocked prior to DPA release and cortex hydrolysis, and germination and viability defects in these spores were complemented by an ectopic cspB. These results indicate that Csp proteases are essential to generate active SleC and allow cortex hydrolysis early in C. perfringens spore germination. However, Csp proteases likely play another role in spore germination, since cspB spores did not release DPA upon exposure to germinants, while sleC spores have been shown previously to release DPA, albeit slowly, upon exposure to germinants.

Clostridium perfringens spore germination: characterization of germinants and their receptors.

Clostridium perfringens food poisoning is caused by type A isolates carrying a chromosomal enterotoxin (cpe) gene (C-cpe), while C. perfringens-associated non-food-borne gastrointestinal (GI) diseases are caused by isolates carrying a plasmid-borne cpe gene (P-cpe). C. perfringens spores are thought to be the important infectious cell morphotype, and after inoculation into a suitable host, these spores must germinate and return to active growth to cause GI disease. We have found differences in the germination of spores of C-cpe and P-cpe isolates in that (i) while a mixture of L-asparagine and KCl was a good germinant for spores of C-cpe and P-cpe isolates, KCl and, to a lesser extent, L-asparagine triggered spore germination in C-cpe isolates only; and (ii) L-alanine or L-valine induced significant germination of spores of P-cpe but not C-cpe isolates. Spores of a gerK mutant of a C-cpe isolate in which two of the proteins of a spore nutrient germinant receptor were absent germinated slower than wild-type spores with KCl, did not germinate with L-asparagine, and germinated poorly compared to wild-type spores with the nonnutrient germinants dodecylamine and a 1:1 chelate of Ca2+ and dipicolinic acid. In contrast, spores of a gerAA mutant of a C-cpe isolate that lacked another component of a nutrient germinant receptor germinated at the same rate as that of wild-type spores with high concentrations of KCl, although they germinated slightly slower with a lower KCl concentration, suggesting an auxiliary role for GerAA in C. perfringens spore germination. In sum, this study identified nutrient germinants for spores of both C-cpe and P-cpe isolates of C. perfringens and provided evidence that proteins encoded by the gerK operon are required for both nutrient-induced and non-nutrient-induced spore germination.

Influence of Several Methodological Factors on the Growth of Clostridium perfringens in Cooling Rate Challenge Studies

Proper temperature control is essential in preventing Clostridium perfringens food poisoning. The U.S. Department of Agriculture Food Safety and Inspection Service cooling guidelines offer two options for the cooling of meat products: follow a standard time-temperature schedule or validate that alternative cooling regimens result in no more than a 1-log CFU/g increase of C. perfringens and no growth of Clostridium botulinum. The latter option requires laboratory challenge studies to validate the efficacy of a given cooling process. Accordingly, the objective of this study was to investigate the role of several methodological variables that might be encountered during typical C. perfringens challenge studies. Variables studied included plastic bag type (Whirlpak or Spiral Biotech), sealing method (Multivac or FoodSaver), initial spore inoculum size (1 to approximately 3 log CFU/g), and growth environment (ground beef or Trypticase–peptone–glucose–yeast extract [TPGY] broth). The major factors that affected growth were sample bag type and growth environment. Samples incubated in Whirlpak bags showed significantly less growth than those incubated in Spiral Biotech bags, which was likely due to the former bag's greater oxygen permeability. C. perfringens spores showed shorter germination, outgrowth, and lag times and C. perfringens cells showed faster growth rates in ground beef compared with TPGY broth. No significant difference was observed between two different sealing methods. Initial spore inoculum levels in the range studied had no significant effect on final C. perfringens cell concentration.

Preformed Bacterial Toxins

Food poisoning syndromes caused by four different bacteria are described. For all types, food kept at a permissive temperature allows growth of the vegetative forms of the bacteria and production of a toxin or toxins. The key features of these syndromes, as well as possible new trends of concern, are summarized in Table 1.

Bacteriological investigation of an outbreak of Clostridium perfringens food poisoning caused by Japanese food without animal protein

An outbreak of Clostridium perfringens food poisoning occurred in a senior citizen’s home in Japan. Japanese food, spinach boiled with fried bean curd, was considered to be the causative food as a result of the detection of the C. perfringens enterotoxin gene by nested PCR. The number of enterotoxin-positive C. perfringens was enumerated as 4.3×10 5/g in the causative food by the MPN method combined with nested PCR. By cultivation, enterotoxin-positive C. perfringens was isolated from all the fecal specimens of patients tested and the causative food. The isolates from patients were serotypable, heat-resistant and the majority produced enterotoxin, however most isolates from the causative food were nonserotypable, enterotoxin-negative and heat-sensitive.

Direct detection of Clostridium perfringens enterotoxin in patients' stools during an outbreak of food poisoning.

An outbreak of diarrhoea in a hotel affected 25 time keepers attending the 1997 Mediterranean Games. Epidemiological investigation implicated a 'pasta al ragù' consumed at the hotel's restaurant and Clostridium perfringens food poisoning was identified by direct detection of C. perfringens enterotoxin in patients' stools. This report confirms that a careful evaluation of epidemiological features, together with the availability of direct and rapid laboratory methods, may lead to a prompt identification of C. perfringens food poisoning.

Direct detection of Clostridium perfringens enterotoxin in patients' stools during an outbreak of food poisoning

An outbreak of diarrhoea in a hotel affected 25 time keepers attending the 1997 Mediterranean Games. Epidemiological investigation implicated a ‘pasta al ragù’ consumed at the hotel's restaurant and Clostridium perfringens food poisoning was identified by direct detection of C. perfringens enterotoxin in patients' stools. This report confirms that a careful evaluation of epidemiological features, together with the availability of direct and rapid laboratory methods, may lead to a prompt identification of C. perfringens food poisioning.

Clostridial disease of the gut.

Clostridia are an important cause of morbidity and mortality in humans and animals. Some of the most common clostridial infections are those of the gut. The primary infections in humans are Clostridium perfringens food poisoning and Clostridium difficile-mediated antibiotic-associated diarrhea and colitis. Less common but important infections include non-food poisoning C. perfringens nosocomial diarrhea and C. perfringens type C necrotizing jejunitis (pig-bel). C. perfringens is also the dominant cause of gastrointestinal infections in animals, although Clostridium septicum causing braxy in sheep, Clostridium colinum causing ulcerative enteritis is avian species, and Clostridium spiroforme causing enterotoxemia in rabbits are important exceptions.

Clostridium perfringens outbreak associated with minestrone soup.

Clostridium perfringens food poisoning usually occurs after animal protein foods such as meat or poultry are consumed. In November 1990, an outbreak of gastrointestinal illness occurred among attendees at a conference of cake decorators held in Michigan. All 42 conference attendees were surveyed. Thirty-two of 42 (76%) attendees reported gastrointestinal symptoms. The median incubation period was 6.5 hours (range, 1.5-15 hours). Ill persons were almost five times more likely to have consumed minestrone soup than were those who were not ill (relative risk = 4.92, 95% confidence interval 1.23-infinity). Bacteriologic analysis identified C. perfringens in the soup and in 11 of 12 stool specimens from ill persons. The soup was cooked 2 days prior to serving and was slowly cooled before refrigeration; it was briefly reheated on the day served. Failure to monitor time and temperature controls during the cooling and reheating of the soup probably allowed for C. perfringens proliferation in the minestrone soup. This investigation demonstrated that nonanimal protein sources, such as legumes, can support substantial C. perfringens growth and should be regarded as potential vehicles in C. perfringens outbreaks.

Outbreak of Clostridium perfringens food poisoning

An outbreak of diarrhoea occurred at a 647-bedded long-stay hospital from 11 to 14 June 1989. Fifty-eight elderly residents developed symptoms and there were two deaths. The organism was identified as Clostridium perfringens type A, serotype TW23. The source of the outbreak was found to be inadequately reheated minced beef served at lunchtime on 11 June. The reason why only 4 of the possible 16 wards receiving minced beef were affected was thought to be due to the division of the batch of minced beef at a late stage in the preparation process. We conclude that there is a need for effective bowel and nutrition policies and that these are high priorities for audit.

Analytical methods for Bacillus cereus and other Bacillus species

Bacillus cereus can give rise to two distinct forms of foodborne disease, the emetic and the diarrhoeal syndromes. The emetic syndrome is believed to be associated with an emetic toxin pre-formed in food. Cooked rice is the most common vehicle, and the symptoms are similar to those of Staphylococcus aureus intoxication. The diarrhoeal type is caused by an enterotoxin and the symptoms generally parallel those of the Clostridium perfringens food poisoning. The heat resistance of B. cereus spores and the non-fastidious nature of the organism facilitates its survival and/or growth in a wide variety of foods. This review describes analytical methods available for the isolation, identification, and enumeration of the organism, in addition to details about biological and immunological methods for toxin assay. Data are also presented concerning the incidence and epidemiology of B. cereus food poisoning around the world, and especially in Japan.

Clostridium perfringens food poisoning: use of serotyping in an outbreak setting

An outbreak of Clostridium perfringens food poisoning occurred among attendees of a firehouse luncheon. The predominant symptoms of diarrhea (100%) and abdominal pain (81%) among case-patients, the mean incubation period (13.4 h), and the mean duration of illness (21.2 h) were all characteristic of C. perfringens enteritis. Roast beef, although not epidemiologically implicated, was the most likely vehicle of transmission. Fecal specimens from case-patients contained a median C. perfringens spore count of greater than 10(6) and yielded isolates that were heat sensitive and predominantly nonhemolytic, produced C. perfringens enterotoxin A, and, in the majority of specimens (four of five), were identical in serotype. Food samples were negative. This outbreak demonstrates that following enumeration of C. perfringens from a suitable number of fecal specimens from case-patients, serotyping of the isolates may be helpful in implicating C. perfringens as the cause of foodborne illness. This is especially true when implicated food items test negative or are no longer available for testing.

Characterization of an outbreak of Clostridium perfringens food poisoning by quantitative fecal culture and fecal enterotoxin measurement.

Published criteria for implicating Clostridium perfringens as the cause of food-poisoning outbreaks include finding a median fecal C. perfringens spore count of greater than 10(6)/g among specimens from ill persons. We investigated a food-poisoning outbreak with the epidemiologic characteristics of C. perfringens-related disease in a nursing home in which the median fecal spore count for ill patients (2.5 X 10(7)/g) was similar to that for well patients (4.0 X 10(6)/g), making the etiology of the outbreak uncertain. All ill and well patients tested had eaten turkey, the implicated food item. C. perfringens enterotoxin was detected by reverse passive latex agglutination in fecal specimens from six of six ill and none of four well patients who had eaten turkey (P = 0.005), suggesting that this organism had caused the outbreak. This investigation suggests that detection of fecal C. perfringens enterotoxin is a specific way to identify this organism as the causative agent in food-poisoning outbreaks.

Comparison of latex agglutination and ELIS A for the detection of Clostridium perfringens type A enterotoxin in faeces

One hundred and thirty one faecal specimens from cases of suspected Clostridium perfringens food poisoning were examined by both a reverse passive latex agglutination test and a standard ELISA test for the presence of Cl. perfringens enterotoxin. The latex agglutination test proved as sensitive and specific as the ELISA, and required less time at the bench without the need for specialized equipment.

The potential of bacteriocin typing in the study of Clostridium perfringens food poisoning.

A range of 49 bacteriocins was used to type 311 strains of Clostridium perfringens isolated from food poisoning outbreaks. Strains of same serotype within an outbreak showed similar patterns of susceptibility to bacteriocins, whereas strains of different serotype isolated from different sources produced many variations in bacteriocin susceptibility patterns. The 311 strains, along with isolates from a wide range of sources were screened for their ability to produce bacteriocins. A much greater proportion of the strains from food poisoning outbreaks was bacteriocinogenic than were isolates from human and animal infections, various foods and the environment.