scholarly journals Enterotoxin Plasmid from Clostridium perfringens Is Conjugative

2001 ◽  
Vol 69 (5) ◽  
pp. 3483-3487 ◽  
Author(s):  
Sigrid Brynestad ◽  
Mahfuzur R. Sarker ◽  
Bruce A. McClane ◽  
Per Einar Granum ◽  
Julian I. Rood
Keyword(s):  
Clostridium Perfringens ◽  
Gastrointestinal Disease ◽  
Intestinal Flora ◽  
Food Poisoning ◽  
Donor Cell ◽  
Cell Contact ◽  
Conjugative Transfer ◽  
Chloramphenicol Resistance ◽  
Food Borne

ABSTRACT Clostridium perfringens enterotoxin is the major virulence factor involved in the pathogenesis of C. perfringens type A food poisoning and several non-food-borne human gastrointestinal illnesses. The enterotoxin gene,cpe, is located on the chromosome of food-poisoning isolates but is found on a large plasmid in non-food-borne gastrointestinal disease isolates and in veterinary isolates. To evaluate whether the cpe plasmid encodes its own conjugative transfer, a C. perfringens strain carrying pMRS4969, a plasmid in which a 0.4-kb segment internal to thecpe gene had been replaced by the chloramphenicol resistance gene catP, was used as a donor in matings with several cpe-negative C. perfringensisolates. Chloramphenicol resistance was transferred at frequencies ranging from 2.0 × 10−2 to 4.6 × 10−4 transconjugants per donor cell. The transconjugants were characterized by PCR, pulsed-field gel electrophoresis, and Southern hybridization analyses. The results demonstrated that the entire pMRS4969 plasmid had been transferred to the recipient strain. Plasmid transfer required cell-to-cell contact and was DNase resistant, indicating that transfer occurred by a conjugation mechanism. In addition, several fragments of the prototype C. perfringens tetracycline resistance plasmid, pCW3, hybridized with pMRS4969, suggesting that pCW3 shares some similarity to pMRS4969. The clinical significance of these findings is that if conjugative transfer of the cpe plasmid occurred in vivo, it would have the potential to convertcpe-negative C. perfringens strains in normal intestinal flora into strains capable of causing gastrointestinal disease.

scholarly journals Complete Sequencing and Diversity Analysis of the Enterotoxin-Encoding Plasmids in Clostridium perfringens Type A Non-Food-Borne Human Gastrointestinal Disease Isolates

2006 ◽  
Vol 188 (4) ◽  
pp. 1585-1598 ◽  
Author(s):  
Kazuaki Miyamoto ◽  
Derek J. Fisher ◽  
Jihong Li ◽  
Sameera Sayeed ◽  
Shigeru Akimoto ◽  
...  
Keyword(s):  
Clostridium Perfringens ◽  
Gastrointestinal Disease ◽  
Food Poisoning ◽  
Variable Region ◽  
Type A ◽  
Gastrointestinal Diseases ◽  
Open Reading Frames ◽  
Toxin Gene ◽  
Virulence Plasmids ◽  
Food Borne

ABSTRACT Enterotoxin-producing Clostridium perfringens type A isolates are an important cause of food poisoning and non-food-borne human gastrointestinal diseases, e.g., sporadic diarrhea (SPOR) and antibiotic-associated diarrhea (AAD). The enterotoxin gene (cpe) is usually chromosomal in food poisoning isolates but plasmid-borne in AAD/SPOR isolates. Previous studies determined that type A SPOR isolate F5603 has a plasmid (pCPF5603) carrying cpe, IS1151, and the beta2 toxin gene (cpb2), while type A SPOR isolate F4969 has a plasmid (pCPF4969) lacking cpb2 and IS1151 but carrying cpe and IS1470-like sequences. By completely sequencing these two cpe plasmids, the current study identified pCPF5603 as a 75.3-kb plasmid carrying 73 open reading frames (ORFs) and pCPF4969 as a 70.5-kb plasmid carrying 62 ORFs. These plasmids share an ∼35-kb conserved region that potentially encodes virulence factors and carries ORFs found on the conjugative transposon Tn916. The 34.5-kb pCPF4969 variable region contains ORFs that putatively encode two bacteriocins and a two-component regulator similar to VirR/VirS, while the ∼43.6-kb pCPF5603 variable region contains a functional cpb2 gene and several metabolic genes. Diversity studies indicated that other type A plasmid cpe +/IS1151 SPOR/AAD isolates carry a pCPF5603-like plasmid, while other type A plasmid cpe +/IS1470-like SPOR/AAD isolates carry a pCPF4969-like plasmid. Tn916-related ORFs similar to those in pCPF4969 (known to transfer conjugatively) were detected in the cpe plasmids of other type A SPOR/AAD isolates, as well as in representative C. perfringens type B to D isolates carrying other virulence plasmids, possibly suggesting that most or all C. perfringens virulence plasmids transfer conjugatively.

scholarly journals Comparative Experiments To Examine the Effects of Heating on Vegetative Cells and Spores of Clostridium perfringens Isolates Carrying Plasmid Genes versus Chromosomal Enterotoxin Genes

2000 ◽  
Vol 66 (8) ◽  
pp. 3234-3240 ◽  
Author(s):  
Mahfuzur R. Sarker ◽  
Robert P. Shivers ◽  
Shauna G. Sparks ◽  
Vijay K. Juneja ◽  
Bruce A. McClane
Keyword(s):  
Clostridium Perfringens ◽  
Gastrointestinal Disease ◽  
Food Poisoning ◽  
Meat Products ◽  
Gastrointestinal Diseases ◽  
Vegetative Cells ◽  
Food Borne ◽  
Enterotoxin Genes ◽  
Plasmid Genes ◽  
High Degree

ABSTRACT Clostridium perfringens enterotoxin (CPE) is an important virulence factor for both C. perfringens type A food poisoning and several non-food-borne human gastrointestinal diseases. Recent studies have indicated that C. perfringensisolates associated with food poisoning carry a chromosomalcpe gene, while non-food-borne human gastrointestinal disease isolates carry a plasmid cpe gene. However, no explanation has been provided for the strong associations between certain cpe genotypes and particular CPE-associated diseases. Since C. perfringens food poisoning usually involves cooked meat products, we hypothesized that chromosomalcpe isolates are so strongly associated with food poisoning because (i) they are more heat resistant than plasmid cpeisolates, (ii) heating induces loss of the cpe plasmid, or (iii) heating induces migration of the plasmid cpe gene to the chromosome. When we tested these hypotheses, vegetative cells of chromosomal cpe isolates were found to exhibit, on average approximately twofold-higher decimal reduction values (Dvalues) at 55°C than vegetative cells of plasmid cpeisolates exhibited. Furthermore, the spores of chromosomalcpe isolates had, on average, approximately 60-fold-higherD values at 100°C than the spores of plasmidcpe isolates had. Southern hybridization and CPE Western blot analyses demonstrated that all survivors of heating retained theircpe gene in its original plasmid or chromosomal location and could still express CPE. These results suggest that chromosomalcpe isolates are strongly associated with food poisoning, at least in part, because their cells and spores possess a high degree of heat resistance, which should enhance their survival in incompletely cooked or inadequately warmed foods.

scholarly journals Organization of the Plasmid cpe Locus in Clostridium perfringens Type A Isolates

2002 ◽  
Vol 70 (8) ◽  
pp. 4261-4272 ◽  
Author(s):  
Kazuaki Miyamoto ◽  
Ganes Chakrabarti ◽  
Yosiharu Morino ◽  
Bruce A. McClane
Keyword(s):  
Clostridium Perfringens ◽  
Horizontal Transfer ◽  
Food Poisoning ◽  
Type A ◽  
Gastrointestinal Diseases ◽  
Open Reading Frame ◽  
Enterotoxin Gene ◽  
Genetic Element ◽  
Reading Frame ◽  
Food Borne

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.

scholarly journals Evaluating the Involvement of Alternative Sigma Factors SigF and SigG in Clostridium perfringens Sporulation and Enterotoxin Synthesis

2010 ◽  
Vol 78 (10) ◽  
pp. 4286-4293 ◽  
Author(s):  
Jihong Li ◽  
Bruce A. McClane
Keyword(s):  
Bacillus Subtilis ◽  
Clostridium Perfringens ◽  
Food Poisoning ◽  
Sigma Factors ◽  
Wild Type ◽  
Alternative Sigma Factors ◽  
Food Borne ◽  
Food Borne Illness ◽  
Bacterial Food ◽  
Null Mutants

ABSTRACT Clostridium perfringens type A food poisoning is the second most commonly identified bacterial food-borne illness. Sporulation contributes to this disease in two ways: (i) most food-poisoning strains form exceptionally resistant spores to facilitate their survival of food-associated stresses, and (ii) the enterotoxin (CPE) responsible for the symptoms of this food poisoning is synthesized only during sporulation. In Bacillus subtilis, four alternative sigma factors mediate sporulation. The same four sigma factors are encoded by C. perfringens genomes, and two (SigE and SigK) have previously been shown to be necessary for sporulation and CPE production by SM101, a transformable derivative of a C. perfringens food-poisoning strain (K. H. Harry, R. Zhou, L. Kroos, and S. B. Melville, J. Bacteriol. 2009, 191:2728-2742). However, the importance of SigF and SigG for C. perfringens sporulation or CPE production had not yet been assessed. In the current study, after confirming that sporulating wild-type SM101 cultures produce SigF (from a tricistronic operon) and SigG, we prepared isogenic sigF- or sigG-null mutants. Whereas SM101 formed heat-resistant, phase-refractile spores, spore formation was blocked in the sigF- and sigG-null mutants. Complementation fully restored sporulation by both mutants. By use of these mutants and complementing strains, CPE production was shown to be SigF dependent but SigG independent. This finding apparently involved regulation of the production of SigE and SigK, which Harry et al. showed to be necessary for CPE synthesis, by SigF. By combining these findings with those previous results, it is now apparent that all four alternative sigma factors are necessary for C. perfringens sporulation, but only SigE, SigF, and SigK are needed for CPE synthesis.

scholarly journals Clostridium perfringens – epidemiological importance and diagnostics

2019 ◽  
Vol 75 (01) ◽  
pp. 6161-2019
Author(s):  
NINA KOZIEŁ ◽  
ELŻBIETA KUKIER ◽  
KRZYSZTOF KWIATEK
Keyword(s):  
Clostridium Perfringens ◽  
Extracellular Enzymes ◽  
Food Poisoning ◽  
Immunocompromised Host ◽  
Necrotic Enteritis ◽  
Virulence Determinants ◽  
Gastrointestinal Infections ◽  
Food Borne ◽  
Disease Diagnostics ◽  
Enteritis Necroticans

Clostridium perfringens is one of the most widespread anaerobic spore forming bacteria found in the environment. The toxotype A of the species inhabits the gastrointestinal tract of birds and mammals exhibiting pathogenic properties in the immunocompromised host. The virulence determinants of C. perfringens are toxins and extracellular enzymes which cause gas gangrene, enteritis necroticans, food poisoning, and non-food borne gastrointestinal infections in humans. The young animals suffer from enterotoxaemia, dysentery and necrotic enteritis due to the anaerobic spore forming bacilli. This article reviews the epidemiological significance of C. perfringens and its disease diagnostics, taking into account all known to date virulence determinants of the microorganism.

scholarly journals Shiga toxin-producing Escherichia coli (STEC) food-borne outbreaks

2017 ◽  
Vol 63 (1) ◽  
pp. 45 ◽  
Author(s):  
A. PEXARA (Α. ΠΕΞΑΡΑ) ◽  
A. S. ANGELIDIS (Α. Σ. ΑΓΓΕΛΙΔΗΣ) ◽  
A. GOVARIS (Α. ΓΚΟΒΑΡΗΣ)
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Intestinal Flora ◽  
Food Poisoning ◽  
Significant Risk ◽  
Public Health Problem ◽  
Human Infection ◽  
Food Sources ◽  
E Coli ◽  
Food Borne

Escherichia coli (E. coli) are Gram negativo, non-sporulating bacteria, which belong to the normal intestinal flora of humans and animals. Shiga toxin-producing E. coli (STFC) arc a group of if. coli that is defined by the capacity to produce toxins called Shiga toxins (Stx). hollowing ingestion of STEC, the significant risk of two serious and potentially life-threatening complications of infection, hemorrhagic colitis and hemolytic uremic syndrome (HUS), makes STHC food poisoning a serious public health problem. Besides Stx, human pathogenic STFC harbor additional virulence factors that are important for their pathogenicity. Although human infection may also be acquired by direct transmission from person to person or by direct contact of humans with animal carriers, the majority of STFC infections are food-borne in origin.The gastrointestinal tract of healthy ruminants seems to be the foremost important reservoir for STFC and ingestion of undercooked beef one of the most likely routes of transmission to humans, Other important food sources include faecally contaminated vegetables and drinking water, The serogroup classification of STHC is based on the somatic (O) and flagellar (H) antigens, and, to date, more than 200 STFC serogroups have been identified, Human STFC infections are, however, associated with a minor subset of 0;H serotypes. Of these, the 0157:H7 or the 0157 :H- serogroups (STFC 0157) are the ones most frequently reported to be associated with food-borne outbreaks. However other non-0157 STFC serogroups such as E. coli 026, 0103, O l l i , 012I, 045 and 0145 have caused several outbreaks in recent years.Two outbreaks of gastroenteritis caused by E. coli 0157:117 were first reported in the US, following the consumption of undercooked hamburgers, in 1982. Since then several outbreaks were reported worldwide. A major E. coli 0157:117 outbreak occurred in Japan and contaminated radish sprouts was identified as the vehicle of infection. More than 6,000 school children were affected, 101 people were hospitalized with lILS and 12 deaths were recorded. The recent outbreak of STFC 0104:114 infection and HUS reported in Germany in the spring of 2011 was one of the largest outbreaks worldwide. As of 27 July, 3 126 cases of STFC infections, 773 cases of HUS including 46 deaths linked to the outbreak in Germany and occurring in the Furopean Union (FU) (including Norway), Outside the FU 8 cases of STFC and 5 cases of HUS, including 1 death have been reported in the USA, Canada and Switzerland, all with recent travel history to Germany.The present review on major STliC food-borne outbreaks recorded worldwide highlights the need for eontrol measures in order to prevent or at least minimize the occurrence of similar events in the future.

scholarly journals ROLE OF CLOSTRIDIUM PERFRINGENS IN PATHOGENICITY OF SOME DOMESTIC ANIMALS

2017 ◽  
Vol 7 (3) ◽  
pp. 1117-1121 ◽  
Author(s):  
Mohammad Reza Mohammadabadi
Keyword(s):  
Clostridium Perfringens ◽  
Gastrointestinal Disease ◽  
Food Poisoning ◽  
Small Ruminants ◽  
Necrotic Enteritis ◽  
Economic Stability ◽  
Native Breed ◽  
The World ◽  
The Tropics

Clostridium perfringens, is an anaerobic, gram-positive, pathogenic and spore-forming bacillus and broadly gave out in our territory. This bacterium has spore formation capability and creating gangrene and gastrointestinal disease, for example food poisoning and necrotic enteritis in human, whilst in other animals, gastrointestinal and enterotoxemic diseases more happening. Prevalence of necrotic enteritis, created by C. perfringens, has been often stated in sheep, chickens and ostrich throughout the world. The most critical problem for epidemiological investigations and vaccines improvement is accurate recognition of C. perfringens variants. Moreover, Small ruminants, especially native breed types, play an important role to the livelihoods of a considerable part of human population in the tropics from socio-economic aspects. Therefore, integrated attempt in terms of management and genetic improvement to enhance production is of crucial importance. Poultry provide humans with companionship, food and fiber in the form of eggs, meat and feathers. Many people love to raise and show chickens and other poultry species at fairs and other poultry shows. Others just love to raise them for backyard pets and for fresh eggs every day. In the last few years, ostrich farming has progressed dramatically and the world ostrich industry has achieved some economic stability. There is considerable scope for improvement in the areas of artificial incubation, chick nutrition, environmental requirements and selective breeding. Hence, the aim of this paper was to study role of Clostridium perfringens in pathogenicity of sheep, broilers and Ostrich. In conclusion, recognition of toxins producing by C. perfringens is very momentous because their toxin types are related to particular gastric and intestinal animal sickness and PCR has become an essential research and diagnostic tool, being a powerful technique with a vast and increasing range of applications. Hence, it is better that animal breeders identify different types of C. perfringens using PCR technique to prevent the damage caused by this bacterium.  

scholarly journals Inorganic Phosphate and Sodium Ions Are Cogerminants for Spores of Clostridium perfringens Type A Food Poisoning-Related Isolates

2009 ◽  
Vol 75 (19) ◽  
pp. 6299-6305 ◽  
Author(s):  
Daniel Paredes-Sabja ◽  
Pathima Udompijitkul ◽  
Mahfuzur R. Sarker
Keyword(s):  
Clostridium Perfringens ◽  
Inorganic Phosphate ◽  
Dipicolinic Acid ◽  
Food Poisoning ◽  
Type A ◽  
Gastrointestinal Diseases ◽  
Wild Type ◽  
Similar Extent ◽  
Active Growth ◽  
Food Borne

ABSTRACT Clostridium perfringens type A isolates carrying a chromosomal copy of the enterotoxin (cpe) gene are involved in the majority of food poisoning (FP) outbreaks, while type A isolates carrying a plasmid-borne cpe gene are involved in C. perfringens-associated non-food-borne (NFB) gastrointestinal diseases. To cause diseases, C. perfringens spores must germinate and return to active growth. Previously, we showed that only spores of FP isolates were able to germinate with K+ ions. We now found that the spores of the majority of FP isolates, but none of the NFB isolates, germinated with the cogerminants Na+ and inorganic phosphate (NaPi) at a pH of ∼6.0. Spores of gerKA-KC and gerAA mutants germinated to a lesser extent and released less dipicolinic acid (DPA) than did wild-type spores with NaPi. Although gerKB spores germinated to a similar extent as wild-type spores with NaPi, their rate of germination was lower. Similarly, gerO and gerO gerQ mutant spores germinated slower and released less DPA than did wild-type spores with NaPi. In contrast, gerQ spores germinated to a slightly lesser extent than wild-type spores but released all of their DPA during NaPi germination. In sum, this study identified NaPi as a novel nutrient germinant for spores of most FP isolates and provided evidence that proteins encoded by the gerKA-KC operon, gerAA, and gerO are required for NaPi-induced spore germination.

scholarly journals Inorganic Phosphate Induces Spore Morphogenesis and Enterotoxin Production in the Intestinal Pathogen Clostridium perfringens

2006 ◽  
Vol 74 (6) ◽  
pp. 3651-3656 ◽  
Author(s):  
Valeria A. Philippe ◽  
Marcelo B. Méndez ◽  
I-Hsiu Huang ◽  
Lelia M. Orsaria ◽  
Mahfuzur R. Sarker ◽  
...  
Keyword(s):  
Clostridium Perfringens ◽  
Inorganic Phosphate ◽  
Food Poisoning ◽  
Inhibitory Effect ◽  
Physiological Signal ◽  
Food Borne ◽  
Intestinal Pathogen ◽  
Important Virulence Factor ◽  
Dna Partitioning ◽  
Effect Of Glucose

ABSTRACT Clostridium perfringens enterotoxin (CPE) is an important virulence factor for food poisoning and non-food borne gastrointestinal (GI) diseases. Although CPE production is strongly regulated by sporulation, the nature of the signal(s) triggering sporulation remains unknown. Here, we demonstrated that inorganic phosphate (Pi), and not pH, constitutes an environmental signal inducing sporulation and CPE synthesis. In the absence of Pi-supplementation, C. perfringens displayed a spo0A phenotype, i.e., absence of polar septation and DNA partitioning in cells that reached the stationary phase of growth. These results received support from our Northern blot analyses which demonstrated that Pi was able to counteract the inhibitory effect of glucose at the onset of sporulation and induced spo0A expression, indicating that Pi acts as a key signal triggering spore morphogenesis. In addition to being the first study reporting the nature of a physiological signal triggering sporulation in clostridia, these findings have relevance for the development of antisporulation drugs to prevent or treat CPE-mediated GI diseases in humans.

scholarly journals Food poisoning in hospitals in Scotland

1979 ◽  
Vol 83 (2) ◽  
pp. 231-236 ◽  
Author(s):  
J. C. M. Sharp ◽  
P. W. Collier ◽  
R. J. Gilbert
Keyword(s):  
Staphylococcus Aureus ◽  
Clostridium Perfringens ◽  
Preventive Measures ◽  
Food Poisoning ◽  
Salmonella Infection ◽  
England And Wales ◽  
Laboratory Facilities ◽  
Food Borne ◽  
Prepared Food

summaryA review of 50 hospital-based outbreaks of food poisoning which were reported in Scotland during 1973–7, is described. At least 1530 persons consuming hospital-prepared food were involved. Thirty-one episodes were associated with Clostridium perfringens (C. welchii), 11 were due to food-borne salmonella infection, three to enterotoxigenic Staphylococcus aureus, and five incidents were of undetermined aetiology. This differs noticeably from the experience in England and Wales where salmonellas appear to predominate as the main cause of hospital outbreaks. Twenty-two incidents occurred in hospitals for psychiatric or mentally subnormal patients, and ten others were located in geriatric units. Only 33 hospitals were involved in the 50 outbreaks as nine hospitals experienced two or more episodes.The role of the hospital in the occurrence of food poisoning may be over-emphasized in comparison with other catering establishments, as outbreaks are more readily recognized and laboratory facilities are usually available for investigation, but it is also believed that many episodes may not be reported. The peculiar problems of the hospital-catering service and particularly those of the older long-stay hospitals, are discussed in relation to preventive measures which would minimize the hazards of food poisoning.

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