scholarly journals A rapid qualitative assay for detection of Clostridium perfringens in canned food products

2017 ◽  
Vol 64 (2) ◽  
Author(s):  
Gayatri Ashwinkumar Dave
Keyword(s):  
Phospholipase C ◽  
Clostridium Perfringens ◽  
Colorimetric Assay ◽  
Food Poisoning ◽  
The United States ◽  
Necrotic Enteritis ◽  
Yellow Colour ◽  
Negative Results ◽  
Reporter Assays ◽  
Food Borne Infections

Clostridium perfringens (MTCC 1349) is a Gram-positive, anaerobic, endospore forming, and rod-shaped bacterium. This bacterium produces a variety of toxins under strict anaerobic environment. C. perfringens can grow at temperatures ranging between 20°C and 50°C. It is the major causetive agent for gas gangrene, cellulitis, septicemia, necrotic enteritis and food poisoning, which are common toxin induced conditions noted in human and animals. C. perfringens can produce produce four major types of toxins that are used for the classification of strains, classified under type A–E. Across the globe many countries, including the United States, are affected by C. perfringens food poisonings where it is ranked as one of the most common causes of food borne infections. To date, no direct one step assay for the detection of C. perfringens has been developed and only few methods are known for accurate detection of C. perfringens. Long detection and incubation time is the major consideration of these reporter assays. The prensent study proposes a rapid and reliable colorimetric assay for the detection of C. perfringens. In principale, this assay detects the para nitrophenyl (yellow colour end product) liberated due to the hydrolysis of paranitrophenyl phosphetidyl choline (PNPC) through phospholipase C (lecithinase). Constitutive secretion of phospholipase C is a charactristic feature of C. perfringens. This assay detects the presence of the extracellular lecithinse through the PNPC impragnated impregnated probe. The probe is impregnated with peranitrophenyl phosphotidyl choline ester, which is colourless substrate used by lecithinase. The designed assay is specific towards PNPC and detectes very small quantites of lecithinase under conditions used. The reaction is substrate specific, no cross reaction was observed upon incubation with other substrates. In addition, this assay gave negative results with other clostridium strains, no cross reactions were observed with other experimental strains like C. tetani, C. botulinum, C. acetobutyricum, Bacillus subtilis, and Escherichia coli. This assay is extramly rapid and provides reliable and reproducible results within one hour of incubation at 37°C.

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 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 An atypical lipoteichoic acid from Clostridium perfringens elicits a broadly cross-reactive and protective immune response

2020 ◽  
Vol 295 (28) ◽  
pp. 9513-9530 ◽  
Author(s):  
Cory Q. Wenzel ◽  
Dominic C. Mills ◽  
Justyna M. Dobruchowska ◽  
Jiri Vlach ◽  
Harald Nothaft ◽  
...  
Keyword(s):  
Clostridium Perfringens ◽  
Vaccine Development ◽  
Food Poisoning ◽  
Laboratory Strain ◽  
Lipoteichoic Acid ◽  
2D Nmr ◽  
Effective Vaccine ◽  
Necrotic Enteritis ◽  
Bacterial Killing ◽  
Whole Cells

Clostridium perfringens is a leading cause of food-poisoning and causes avian necrotic enteritis, posing a significant problem to both the poultry industry and human health. No effective vaccine against C. perfringens is currently available. Using an antiserum screen of mutants generated from a C. perfringens transposon-mutant library, here we identified an immunoreactive antigen that was lost in a putative glycosyltransferase mutant, suggesting that this antigen is likely a glycoconjugate. Following injection of formalin-fixed whole cells of C. perfringens HN13 (a laboratory strain) and JGS4143 (chicken isolate) intramuscularly into chickens, the HN13-derived antiserum was cross-reactive in immunoblots with all tested 32 field isolates, whereas only 5 of 32 isolates were recognized by JGS4143-derived antiserum. The immunoreactive antigens from both HN13 and JGS4143 were isolated, and structural analysis by MALDI-TOF-MS, GC-MS, and 2D NMR revealed that both were atypical lipoteichoic acids (LTAs) with poly-(β1→4)-ManNAc backbones substituted with phosphoethanolamine. However, although the ManNAc residues in JGS4143 LTA were phosphoethanolamine-modified, a few of these residues were instead modified with phosphoglycerol in the HN13 LTA. The JGS4143 LTA also had a terminal ribose and ManNAc instead of ManN in the core region, suggesting that these differences may contribute to the broadly cross-reactive response elicited by HN13. In a passive-protection chicken experiment, oral challenge with C. perfringens JGS4143 lead to 22% survival, whereas co-gavage with JGS4143 and α-HN13 antiserum resulted in 89% survival. This serum also induced bacterial killing in opsonophagocytosis assays, suggesting that HN13 LTA is an attractive target for future vaccine-development studies.

Food Poisoning Due to Clostridium perfringens in the United States

1983 ◽  
Vol 147 (1) ◽  
pp. 167-170 ◽  
Author(s):  
W. X. Shandera ◽  
C. O. Tacket ◽  
P. A. Blake
Keyword(s):  
United States ◽  
Clostridium Perfringens ◽  
Food Poisoning ◽  
The United States

scholarly journals NanH Is Produced by Sporulating Cultures of Clostridium perfringens Type F Food Poisoning Strains and Enhances the Cytotoxicity of C. perfringens Enterotoxin

mSphere ◽  
2021 ◽  
Vol 6 (2) ◽  
Author(s):  
Jihong Li ◽  
Bruce A. McClane
Keyword(s):  
Clostridium Perfringens ◽  
Virulence Factor ◽  
Mother Cell ◽  
Food Poisoning ◽  
Mature Spore ◽  
The United States ◽  
Content Type ◽  
Intestinal Pathology ◽  
Mother Cells

ABSTRACT Clostridium perfringens type F food poisoning (FP) strains cause one of the most common foodborne illnesses. This FP develops when type F FP strains sporulate in the intestines and produce C. perfringens enterotoxin (CPE), which is responsible for the diarrhea and abdominal cramps of this disease. While C. perfringens can produce up to three different sialidases, the current study surveyed FP strains, which confirmed the results of a previous study that they consistently carry the nanH sialidase gene, often as their only sialidase gene. NanH production was found to be associated with sporulating cultures of the surveyed type F FP strains, including SM101 (a transformable derivative of a FP strain). The sporulation-associated regulation of NanH production by strain SM101 growing in modified Duncan-Strong medium (MDS) was shown to involve Spo0A, but it did not require the completion of sporulation. NanH production was not necessary for either the growth or sporulation of SM101 when cultured in MDS. In those MDS cultures, NanH accumulated in the sporulating mother cell until it was released coincidently with CPE. Since CPE becomes extracellular when mother cells lyse to release their mature spores, this indicates that mother cell lysis is also important for NanH release. The copresence of NanH and CPE in supernatants from lysed sporulating cultures was shown to enhance CPE cytotoxicity for Caco-2 cells. This enhancement was attributable to NanH increasing CPE binding and could be replicated with purified recombinant NanH. These in vitro findings suggest that NanH may be an accessory virulence factor during type F FP. IMPORTANCE Clostridium perfringens type F strains cause the second most common bacterial foodborne illness in the United States. C. perfringens enterotoxin (CPE) is responsible for the diarrhea and cramping symptoms of this food poisoning (FP). Previous studies showed that NanI sialidase can enhance CPE activity in vitro. While many type F FP strains do not produce NanI, they do consistently make NanH sialidase. This study shows that, like CPE, NanH is produced by sporulating type F FP strains and then released extracellularly when their sporulating cells lyse to release their mature spore. NanH was shown to enhance CPE cytotoxicity in vitro by increasing CPE binding to cultured Caco-2 cells. This enhancement could be important because many type F FP strains produce less CPE than necessary (in a purified form) to cause intestinal pathology in animal models. Therefore, NanH represents a potential accessory virulence factor for type F FP.

scholarly journals Development and Application of a Mouse Intestinal Loop Model To Study theIn VivoAction of Clostridium perfringens Enterotoxin

2011 ◽  
Vol 79 (8) ◽  
pp. 3020-3027 ◽  
Author(s):  
Justin A. Caserta ◽  
Susan L. Robertson ◽  
Juliann Saputo ◽  
Archana Shrestha ◽  
Bruce A. McClane ◽  
...  
Keyword(s):  
Clostridium Perfringens ◽  
Cell Damage ◽  
Gastrointestinal Symptoms ◽  
Food Poisoning ◽  
The United States ◽  
Intestinal Lumen ◽  
Intestinal Loop ◽  
Loop Model ◽  
Internal Organs ◽  
Content Type

ABSTRACTClostridium perfringensenterotoxin (CPE) is responsible for causing the gastrointestinal symptoms ofC. perfringenstype A food poisoning, the second most commonly identified bacterial food-borne illness in the United States. CPE is produced by sporulatingC. perfringenscells in the small intestinal lumen, where it then causes epithelial cell damage and villous blunting that leads to diarrhea and cramping. Those effects are typically self-limiting; however, severe outbreaks of this food poisoning, particularly two occurring in psychiatric institutions, have involved deaths. Since animal models are currently limited for the study of the CPE action, a mouse ligated intestinal loop model was developed. With this model, significant lethality was observed after 2 h in loops receiving an inoculum of 100 or 200 μg of CPE but not using a 50-μg toxin inoculum. A correlation was noted between the overall intestinal histological damage and lethality in mice. Serum analysis revealed a dose-dependent increase in serum CPE and potassium levels. CPE binding to the liver and kidney was detected, along with elevated levels of potassium in the serum. These data suggest that CPE can be absorbed from the intestine into the circulation, followed by the binding of the toxin to internal organs to induce potassium leakage, which can cause death. Finally, CPE pore complexes similar to those formed in tissue culture cells were detected in the intestine and liver, suggesting that (i) CPE actions are similarin vivoandin vitroand (ii) CPE-induced potassium release into blood may result from CPE pore formation in internal organs such as the liver.

Enumeration and Characterization of Clostridium perfringens Spores in the Feces of Food Poisoning Patients and Normal Controls

1986 ◽  
Vol 49 (1) ◽  
pp. 23-28 ◽  
Author(s):  
STANLEY M. HARMON ◽  
DONALD A. KAUTTER ◽  
CHARLES L. HATHEWAY
Keyword(s):  
Clostridium Perfringens ◽  
Food Poisoning ◽  
Egg Yolk ◽  
The United States ◽  
Enzyme Linked Immunosorbent Assay ◽  
Minimal Reduction ◽  
Cooked Meat ◽  
Predominant Strain ◽  
Normal Controls

The fecal spore enumeration method for confirming Clostridium perfringens as the cause of food poisoning was evaluated using strains implicated in nine outbreaks in the United States. Confirmed spore counts from 66 stool specimens were made on tryptose-sulfite-cycloserine (TSC) without egg yolk and trypticase soy-sheep blood (TSB) agars. Counts from outbreak stools on TSC agar ranged from 2.0 × 104 to 3.5 × 108 (mean ≥ 1.4 × 106/g) as compared with <103 to 5.0 × 105/g (overall mean 9.5 × 103/g) from normal stools. Similar results were obtained with TSB agar. Isolates from seven of the nine outbreaks were nonhemolytic and produced ≥100 ng enterotoxin/ml in spore broth, as measured by an enzyme-linked immunosorbent assay. Spores in stools from six of the outbreaks were heat-resistant and survived heating for 30 to 60 min at 100°C in cooked meat medium. Strains from the three remaining outbreaks were heat-sensitive and survived heating for only 15 min at 100°C. Enterotoxigenic isolates from all but one of the outbreaks were serotyped. In all instances, the predominant strain in specimens from an outbreak was of the same serotype, indicating that it was the causative strain. Reexamination of five specimens from each of three outbreaks after storage at −20°C for 6 months showed only a minimal reduction in the spore counts.

scholarly journals Clostridium Perfringens Bacteremia with Acute Hemolytic Anemia in the Setting of Endometrial Malignancy

2019 ◽  
Vol 9 (1-2) ◽  
pp. 127-133
Author(s):  
Luke R. Cypher ◽  
Christopher Sullivan ◽  
Ryan Jones ◽  
Angelina Phillips
Keyword(s):  
Phospholipase C ◽  
Hemolytic Anemia ◽  
Clostridium Perfringens ◽  
Rare Case ◽  
Food Poisoning ◽  
Gas Gangrene ◽  
High Grade ◽  
Alpha Toxin ◽  
Intravascular Hemolysis ◽  
Poorly Differentiated

Acute intravascular hemolysis is a rare and often lethal complication of Clostridium perfringens septicemia.Clostridium perfringens is an anaerobic, gram-positive spore-forming rod which is commonly implicated in cases of food poisoning, gas gangrene, and severe hemolytic anemia in humans via the alpha-toxin (phospholipase C). We report an interesting and rare case of a 72-year-old woman who developed massive intravascular hemolysis secondary to C perfringens bacteremia in the setting of poorly differentiated high-grade endometrial malignancy.

Clostridium perfringens

1991 ◽  
Vol 74 (4) ◽  
pp. 711-714 ◽  
Author(s):  
Ronald G Labbé
Keyword(s):  
Clostridium Perfringens ◽  
Food Poisoning ◽  
Food Service ◽  
Receptor Site ◽  
The United States ◽  
Farm Animals ◽  
Enterotoxin Gene ◽  
Elderly Individuals ◽  
Poultry Products ◽  
Healthy Elderly

Abstract In the United States and Canada, Clostridium perfringens remains a leading cause of bacterial food poisoning in humans. It has been primarily associated with meat and poultry products prepared in food service establishments. Fecal spore levels of 104 or more per g are considered indicative of a food poisoning outbreak. However, elevated spore levels of this organism are frequently seen in healthy elderly individuals, an observation that complicates investigations of suspected outbreaks. Recent studies with this population indicate that fecal enterotoxin levels are a valuable and effective assay for confirming outbreaks due to this organism. With regard to the toxin itself, a membrane protein of 50 000-70 000 molecular weight has been isolated as a possible enterotoxin-receptor site. It is the subsequent action of the toxin on membrane structure that results in the loss of ions and fluid associated with illness. In addition, the enterotoxin gene has been cloned in E. coil and sequenced. Using toxin-specific DNA probes, only 6% of non-symptomatic farm animals were found to possess the enterotoxin gene, disproving the hypothesis that all strains of this organism can produce the toxin.

scholarly journals Sporulation and Enterotoxin (CPE) Synthesis Are Controlled by the Sporulation-Specific Sigma Factors SigE and SigK in Clostridium perfringens

2009 ◽  
Vol 191 (8) ◽  
pp. 2728-2742 ◽  
Author(s):  
Kathryn H. Harry ◽  
Ruanbao Zhou ◽  
Lee Kroos ◽  
Stephen B. Melville
Keyword(s):  
Rna Polymerase ◽  
Clostridium Perfringens ◽  
Mother Cell ◽  
Intestinal Tract ◽  
Developmental Process ◽  
Food Poisoning ◽  
The United States ◽  
Sigma Factors ◽  
Gene Encoding ◽  
Promoter Recognition

ABSTRACT Clostridium perfringens is the third most frequent cause of bacterial food poisoning annually in the United States. Ingested C. perfringens vegetative cells sporulate in the intestinal tract and produce an enterotoxin (CPE) that is responsible for the symptoms of acute food poisoning. Studies of Bacillus subtilis have shown that gene expression during sporulation is compartmentalized, with different genes expressed in the mother cell and the forespore. The cell-specific RNA polymerase sigma factors σF, σE, σG, and σK coordinate much of the developmental process. The C. perfringens cpe gene, encoding CPE, is transcribed from three promoters, where P1 was proposed to be σK dependent, while P2 and P3 were proposed to be σE dependent based on consensus promoter recognition sequences. In this study, mutations were introduced into the sigE and sigK genes of C. perfringens. With the sigE and sigK mutants, gusA fusion assays indicated that there was no expression of cpe in either mutant. Results from gusA fusion assays and immunoblotting experiments indicate that σE-associated RNA polymerase and σK-associated RNA polymerase coregulate each other's expression. Transcription and translation of the spoIIID gene in C. perfringens were not affected by mutations in sigE and sigK, which differs from B. subtilis, in which spoIIID transcription requires σE-associated RNA polymerase. The results presented here show that the regulation of developmental events in the mother cell compartment of C. perfringens is not the same as that in B. subtilis and Clostridium acetobutylicum.

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