Production of small, acid-soluble spore proteins inClostridium perfringensnonfoodborne gastrointestinal disease isolates

2007 ◽  
Vol 53 (4) ◽  
pp. 514-518 ◽  
Author(s):  
Deepa Raju ◽  
Mahfuzur R. Sarker
Keyword(s):  
Heat Resistance ◽  
Clostridium Perfringens ◽  
Molecular Basis ◽  
Gastrointestinal Disease ◽  
Food Borne ◽  
Spore Proteins ◽  
The Difference ◽  
Encoding Genes ◽  
Glucuronidase Assay

The molecular basis for the differences in heat resistance between spores of Clostridium perfringens food-borne versus nonfoodborne isolates remains unknown. Since a recent study demonstrated the role of small, acid-soluble spore proteins (SASPs) in heat resistance of spores of food-borne isolates, in the current study, we evaluated the expression of SASP-encoding genes (ssp) and the production of SASPs in nonfoodborne isolates. Our results demonstrated the presence of all three ssp genes in five surveyed nonfoodborne isolates. A β-glucuronidase assay showed that these ssp genes are expressed specifically during sporulation. Furthermore, nonfoodborne isolate F4969 produced SASPs at a level similar to that of food-borne isolate SM101. Collectively, these results suggest that the difference in the levels of heat resistance between spores of food-borne and the nonfoodborne isolates is not the result of impaired expression of ssp genes and (or) decreased production of SASPs in nonfoodborne isolates.

scholarly journals Comparison of the Levels of Heat Resistance of Wild-Type, cpe Knockout, and cpe Plasmid-Cured Clostridium perfringens Type A Strains

2005 ◽  
Vol 71 (11) ◽  
pp. 7618-7620 ◽  
Author(s):  
Deepa Raju ◽  
Mahfuzur R. Sarker
Keyword(s):  
Heat Resistance ◽  
Clostridium Perfringens ◽  
Gastrointestinal Disease ◽  
Type A ◽  
Heat Sensitivity ◽  
Wild Type ◽  
Food Borne ◽  
High Level

ABSTRACT An enterotoxin (cpe) plasmid was cured from a Clostridium perfringens non-food-borne gastrointestinal disease (NFBGID) isolate, and the heat resistance levels of wild-type, cpe knockout, and cpe plasmid-cured strains were compared. Our results demonstrated that (i) wild-type cpe has no influence in mediating high-level heat resistance in C. perfringens and (ii) the cpe plasmid does not confer heat sensitivity on NFBGID isolates.

scholarly journals Multiple-Locus Sequence Typing and Analysis of Toxin Genes in Bacillus cereus Food-Borne Isolates

2007 ◽  
Vol 74 (3) ◽  
pp. 850-860 ◽  
Author(s):  
Barbara Cardazzo ◽  
Enrico Negrisolo ◽  
Lisa Carraro ◽  
Leonardo Alberghini ◽  
Tomaso Patarnello ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Bacillus Cereus ◽  
Reticulate Evolution ◽  
Sequence Information ◽  
Data Set ◽  
Group I ◽  
Food Borne ◽  
Public Data ◽  
Encoding Genes

ABSTRACT In the present study we characterized 47 food-borne isolates of Bacillus cereus using multilocus sequence typing (MLST). Newly determined sequences were combined with sequences available in public data banks in order to produce the largest data set possible. Phylogenetic analysis was performed on a total of 296 strains for which MLST sequence information is available, and three main lineages—I, II, and III—within the B. cereus complex were identified. With few exceptions, all food-borne isolates were in group I. The occurrence of horizontal gene transfer (HGT) among various strains was analyzed by several statistical methods, providing evidence of widespread lateral gene transfer within B. cereus. We also investigated the occurrence of toxin-encoding genes, focusing on their evolutionary history within B. cereus. Several patterns were identified, indicating a pivotal role of HGT in the evolution of toxin-encoding genes. Our results indicate that HGT is an important element in shaping the population structure of the B. cereus complex. The results presented here also provide strong evidence of reticulate evolution within the B. cereus complex.

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 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.

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 Isolation and genotyping of Clostridium perfringens from goats in Minas Gerais, Brazil

2018 ◽  
Vol 48 (7) ◽  
Author(s):  
Jordana Almeida Santana ◽  
Ana Carolina de Andrade Ferreira ◽  
Marina de Castro Campos de Souza ◽  
Maria Aparecida Scatamburlo Moreira ◽  
Magna Coroa Lima ◽  
...  
Keyword(s):  
Clostridium Perfringens ◽  
Small Ruminants ◽  
Type A ◽  
Laboratory Diagnosis ◽  
Minas Gerais ◽  
Stool Samples ◽  
Genotype C ◽  
Type C ◽  
Encoding Genes

ABSTRACT: Despite the known importance of Clostridium perfringens as an enteropathogen in small ruminants, little is known about the role of its additional virulence factors or the frequency of the various C. perfringens genotypes in healthy goats; this complicates the laboratory diagnosis of the infections caused by this microorganism. In light of this, the aim of the present study was to isolate and genotype C. perfringens from stool samples from healthy goats in Brazil. Stool samples from 250 apparently healthy adult goats from 17 different herds in Minas Gerais, Brazil were collected, and isolation and genotyping of C. perfringens was performed. C. perfringens type A was isolated from 189 (75.6%) goats, whereas C. perfringens types C and D were each detected in one goat (0.4%). All isolates were negative for enterotoxin-, NetB-, NetE-, and NetF-encoding genes. These results confirmed C. perfringens type A as part of the microbiota in these animals, and they suggested that C. perfringens type C and D are rarely isolated from healthy goats.

scholarly journals Clostridium perfringens: Sporulation, Spore Resistance and Germination

1970 ◽  
Vol 24 (1) ◽  
pp. 1-8 ◽  
Author(s):  
I-Hsiu Huang ◽  
Deepa Raju ◽  
Daniel Paredes-Sabja ◽  
Mahfuzur R Sarker
Keyword(s):  
United States ◽  
Clostridium Perfringens ◽  
Anaerobic Bacteria ◽  
The United States ◽  
Genome Sequences ◽  
Enteric Diseases ◽  
Spore Resistance ◽  
Food Borne ◽  
Food Borne Illness ◽  
Spore Proteins

Clostridium perfringens are Gram-positive, endospore-forming, anaerobic bacteria with the ability to cause enteric diseases both in human and domesticated animals. As one of the leading cause of food-borne illness in the United States, certain C. perfringens type A isolates exert their action through the production of C. perfringens enterotoxin (CPE), which is expressed only during spore formation. In addition, C. perfringens spores are highly resistant to heat and other environmental factors. Since genome sequences of three C. perfringens strains have been annotated and made public, efforts have been made towards understanding the initiation of sporulation and identifying the key differences between Clostridium and Bacillus sporulation phosphorelay. Small, acid soluble spore proteins (SASPs) have been shown to be required for resistance of C. perfringens spores to heat. Work is also underway to identify nutrient signals required for C. perfringens spore germination. Keywords: Clostridium perfringens, Endospore, Small, acid soluble spore protein (SASP), Heat resistance, GerminationDOI: http://dx.doi.org/10.3329/bjm.v24i1.1229 Bangladesh J Microbiol, Volume 24, Number 1, June 2007, pp 1-8

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 Cysteine-Scanning Mutagenesis Supports the Importance of Clostridium perfringens Enterotoxin Amino Acids 80 to 106 for Membrane Insertion and Pore Formation

2012 ◽  
Vol 80 (12) ◽  
pp. 4078-4088 ◽  
Author(s):  
Jianwu Chen ◽  
James R. Theoret ◽  
Archana Shrestha ◽  
James G. Smedley ◽  
Bruce A. McClane
Keyword(s):  
Amino Acids ◽  
Clostridium Perfringens ◽  
Pore Formation ◽  
Gastrointestinal Symptoms ◽  
Amino Acid Residues ◽  
Content Type ◽  
Food Borne ◽  
Food Borne Illness ◽  
Cysteine Scanning Mutagenesis

ABSTRACTClostridium perfringensenterotoxin (CPE) causes the gastrointestinal symptoms of the second most common bacterial food-borne illness. Previous studies suggested that a region named TM1, which has amphipathic characteristics and spans from amino acids 81 to 106 of the native CPE protein, forms a β-hairpin involved in β-barrel pore formation. To further explore the potential role of TM1 in pore formation, the single Cys naturally present in CPE at residue 186 was first altered to alanine by mutagenesis; the resultant rCPE variant, named C186A, was shown to retain cytotoxic properties. Cys-scanning mutagenesis was then performed in which individual Cys mutations were introduced into each TM1 residue of the C186A variant. When those Cys variants were characterized, three variants were identified that exhibit reduced cytotoxicity despite possessing binding and oligomerization abilities similar to those of the C186A variant from which they were derived. Pronase challenge experiments suggested that the reduced cytotoxicity of those two Cys variants, i.e., the F91C and F95C variants, which model to the tip of the β-hairpin, was attributable to a lessened ability of these variants to insert into membranes after oligomerization. In contrast, another Cys variant, i.e., the G103C variant, with impaired cytotoxicity apparently inserted into membranes after oligomerization but could not form a pore with a fully functional channel. Collectively, these results support the TM1 region forming a β-hairpin as an important step in CPE insertion and pore formation. Furthermore, this work identifies the first amino acid residues specifically involved in those two steps in CPE action.

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.

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