Characterization of enterotoxin producing Clostridium perfringens isolated from foods of animal origin

2017 ◽  
Author(s):  
Shahnaz Haque ◽  
Rajeev Kumar Sharma ◽  
Nagendra Nath Barman ◽  
Laishram Babita Devi ◽  
Luit Moni Barkalita ◽  
...  
Keyword(s):  
Clostridium Perfringens ◽  
Protein Profile ◽  
Meat Products ◽  
Type A ◽  
Animal Origin ◽  
Cytopathic Effects ◽  
Vero Cell Line ◽  
Sds Page ◽  
Type C

Screening of 105 samples comprising of milk, meat and their products for isolation of Clostridium perfringens revealed a total of 39 (37.14%) samples to be positive for C. perfringens, yielding an equal number of isolates. Among the total isolates, 11 (20.0%) belonged to milk and milk products, of which 10 were isolated from raw cow’s milk. The remaining isolate was from paneer sample. Similarly, 28 (56.0%) C. perfringens isolates were recovered from meat and meat products. Majority (37) of the isolates belonged to toxin type A while two isolates recovered each from raw beef and chicken meats were of toxin type C. Among the 39 isolates of C. perfringens, only 5(12.82%) isolates could reveal the presence of cpe (enterotoxin) gene. The protein profile (SDS-PAGE) of both the crude and partially purified enterotoxins prepared from cpe positive C. perfringens isolates (type A and C) revealed a very little difference. Both the partially purified enterotoxins of type A and C of C. perfringens displayed almost similar type of cytopathic effects (CPE) and death of cells in Vero cell line.

Preliminary screening for toxin genes amongst stock cultures of Clostridium perfringens strains isolated from dogs and calves

2014 ◽  
Vol 4 (3) ◽  
pp. 127-132
Author(s):  
Egwari L. O. ◽  
Oghogho E. S. ◽  
Okwumabua O. E. ◽  
Oniha M. I.
Keyword(s):  
Clostridium Perfringens ◽  
Protein Profile ◽  
Animal Origin ◽  
Cell Protein ◽  
Toxin Gene ◽  
Whole Cell ◽  
University Of Wisconsin ◽  
Gene Type ◽  
Whole Cell Protein ◽  
Gene 4

The spectrum of Clostridium perfringens infections ranges from food toxinosis to myonecrosis. In the current study, whole cell protein and toxin gene types were profiled in 12 randomly selected C. perfringens veterinary stock cultures from the University of Wisconsin, Madison to determine epidemio-logical similarity, or diversity amongst strains of animal origin. Whole cell protein analysis was done by SDS-PAGE while toxin gene typing was achieved by extracting DNA by boiling, DNA concentration and purity was determined by spectrophotometer and nanodrop while separation was carried out by checking it on gel electrophoresis. Multiplex PCR was used to identify the toxigenic gene-type. C. perfringens B and C. perfringens EE with established profiles were used as control strains. Isolates typed included strains cp 296, 309, 12872 (from dogs) and 304, 305, 306, 341, 342, 10754, 12218-2, 12218-3, 12473 (from calves). All 12 strains possess the cpa gene, 4 strains have cpb2, 3 strains etx, 2 strains positive for cpe and 1 for cpb. None of the strains carries the itx gene. Two strains have only cpa gene however no strains has more than two toxin gene types, with cpa-cpb2 combination be-ing more frequent. C. perfringens 305 (etx and cpa) and 342 (cpe and cpa) shared the same protein profile but belong to different toxinotype. It is evi-dent that the cpa gene is a marker for all C. perfringens strains, and similarity in protein profile is not sine qua non for toxin gene type.

scholarly journals Genotypic and Phenotypic Characterization of Clostridium perfringens Isolates from Darmbrand Cases in Post-World War II Germany

2012 ◽  
Vol 80 (12) ◽  
pp. 4354-4363 ◽  
Author(s):  
Menglin Ma ◽  
Jihong Li ◽  
Bruce A. McClane
Keyword(s):  
World War Ii ◽  
Heat Resistance ◽  
Clostridium Perfringens ◽  
Type A ◽  
World War ◽  
Content Type ◽  
Post World War Ii ◽  
Heat Resistant ◽  
Type C ◽  
Beta Toxin

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.

scholarly journals Epsilon-Toxin Plasmids of Clostridium perfringens Type D Are Conjugative

2007 ◽  
Vol 189 (21) ◽  
pp. 7531-7538 ◽  
Author(s):  
Meredith L. Hughes ◽  
Rachael Poon ◽  
Vicki Adams ◽  
Sameera Sayeed ◽  
Juliann Saputo ◽  
...  
Keyword(s):  
Clostridium Perfringens ◽  
Mdck Cells ◽  
Type A ◽  
Department Of Agriculture ◽  
Chloramphenicol Resistance ◽  
Cytopathic Effects ◽  
Type D ◽  
Class B ◽  
Epsilon Toxin ◽  
Domestic Livestock

ABSTRACT Isolates of Clostridium perfringens type D produce the potent epsilon-toxin (a CDC/U.S. Department of Agriculture overlap class B select agent) and are responsible for several economically significant enterotoxemias of domestic livestock. It is well established that the epsilon-toxin structural gene, etx, occurs on large plasmids. We show here that at least two of these plasmids are conjugative. The etx gene on these plasmids was insertionally inactivated using a chloramphenicol resistance cassette to phenotypically tag the plasmid. High-frequency conjugative transfer of the tagged plasmids into the C. perfringens type A strain JIR325 was demonstrated, and the resultant transconjugants were shown to act as donors in subsequent mating experiments. We also demonstrated the transfer of “unmarked” native ε-toxin plasmids into strain JIR325 by exploiting the high transfer frequency. The transconjugants isolated in these experiments expressed functional ε-toxin since their supernatants had cytopathic effects on MDCK cells and were toxic in mice. Using the widely accepted multiplex PCR approach for toxin genotyping, these type A-derived transconjugants were genotypically type D. These findings have significant implications for the C. perfringens typing system since it is based on the toxin profile of each strain. Our study demonstrated the fluid nature of the toxinotypes and their dependence upon the presence or absence of toxin plasmids, some of which have for the first time been shown to be conjugative.

Comparative analysis of extractable proteins from Clostridium perfringens type A and type C strains showing varying degree of virulence

Anaerobe ◽  
2015 ◽  
Vol 35 ◽  
pp. 77-91 ◽  
Author(s):  
Pratistha Dwivedi ◽  
Syed Imteyaz Alam ◽  
Om Kumar ◽  
Ravi Bhushan Kumar
Keyword(s):  
Comparative Analysis ◽  
Clostridium Perfringens ◽  
Type A ◽  
Type C

scholarly journals Morphology, Protein Profile and the Level of Infestation of Bedbugs (Hempitera: Cimicidae) in the Halls of Residence of a Higher Institution in Lagos.

2019 ◽  
Vol 1 ◽  
pp. 96-104
Author(s):  
K A Kemabonta ◽  
M O Ajiboye
Keyword(s):  
Staining Method ◽  
Profile Analysis ◽  
Protein Profile ◽  
Polyacrylamide Gel Electrophoresis ◽  
Morphological Characterization ◽  
Sds Page ◽  
Higher Institution ◽  
Cimex Hemipterus ◽  
Very High

Morphology, protein profile and the level of Bedbug infestations were carried out at a University in Lagos, Nigeria after an outcry of high infestation by the students in halls of residence. Three bedbugswere each collected from nine halls of residence for morphological characterization of adults and to carry out protein profile analysis usingSodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) and Coomassie staining method. Level of infestation of Bedbugs was carried out in two female halls A and Busing blood smeared on the walls. All bedbugs found were Cimex hemipterus. 29.8% of the rooms in Hall Bhad no bedbug infestation, 23.9%, 20.2%, 15.4% and 10.6% had low, average, high and very high levels of bedbug infestation respectively while in A hall, it was 21.4%, 30.5%, 24.4%, 14.5% and 9.2% respectively. In A hall, 4.6%, 16.8% and 0.8% had one, two and three out of the four mattresses in the room infested with bedbugs respectively while 56.5% had all the four mattresses infested with bedbugs. In B hall, 1.6%, 16.0% and 0% had one, two and three mattresses infested with bedbugs respectively while the remaining 52.7% had all the four mattresses in the room infested with bedbugs. There was a significant relationship between the level of infestation and the number of mattresses infested in each hall (P < 0.01).The protein profile analysis of bedbugs did not show the protein bands clearly because of the low soluble protein content of Cimex hemipterus and the detection limit of Coomassie stain.

scholarly journals Characterization of KPC-Encoding Plasmids from Enterobacteriaceae Isolated in a Czech Hospital

2017 ◽  
Vol 62 (3) ◽  
Author(s):  
Rudolf Kukla ◽  
Katerina Chudejova ◽  
Costas C. Papagiannitsis ◽  
Matej Medvecky ◽  
Katerina Habalova ◽  
...  
Keyword(s):  
Multidrug Resistance ◽  
Type A ◽  
Type B ◽  
Content Type ◽  
Type C ◽  
Derivatives Of

ABSTRACT Ten Enterobacteriaceae isolates collected in a Czech hospital carried bla KPC -positive plasmids of different sizes (∼30, ∼45, and ∼80 kb). Sequencing revealed three types of plasmids (A to C) with the Tn 4401a transposon. Type A plasmids comprised an IncR backbone and a KPC-2-encoding multidrug resistance (MDR) region. Type B plasmids were derivatives of type A plasmids carrying an IncN3-like segment, while type C plasmids were IncP6 plasmids sharing the same KPC-2-encoding MDR region with type A and B plasmids.

scholarly journals Genetic Characterization of Type A Enterotoxigenic Clostridium perfringens Strains

PLoS ONE ◽  
2009 ◽  
Vol 4 (5) ◽  
pp. e5598 ◽  
Author(s):  
Agi Deguchi ◽  
Kazuaki Miyamoto ◽  
Tomomi Kuwahara ◽  
Yasuhiro Miki ◽  
Ikuko Kaneko ◽  
...  
Keyword(s):  
Clostridium Perfringens ◽  
Genetic Characterization ◽  
Type A
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