Clostridium perfringens Type E Animal Enteritis Isolates with Highly Conserved, Silent Enterotoxin Gene Sequences

ABSTRACT Several Clostridium perfringens genotype E isolates, all associated with hemorrhagic enteritis of neonatal calves, were identified by multiplex PCR. These genotype E isolates were demonstrated to express α and ι toxins, but, despite carrying sequences for the gene (cpe) encoding C. perfringens enterotoxin (CPE), were unable to express CPE. These silent cpe sequences were shown to be highly conserved among type E isolates. However, relative to the functionalcpe gene of type A isolates, these silent type Ecpe sequences were found to contain nine nonsense and two frameshift mutations and to lack the initiation codon, promoters, and ribosome binding site. The type E animal enteritis isolates carrying these silent cpe sequences do not appear to be clonally related, and their silent type E cpe sequences are always located, near the ι toxin genes, on episomal DNA. These findings suggest that the highly conserved, silent cpe sequences present in most or all type E isolates may have resulted from the recent horizontal transfer of an episome, which also carries ι toxin genes, to several different type A C. perfringens isolates.

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

Infection and Immunity ◽

10.1128/iai.70.8.4261-4272.2002 ◽

2002 ◽

Vol 70 (8) ◽

pp. 4261-4272 ◽

Cited By ~ 51

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.

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Clostridium perfringens Type A Strains Carrying a Plasmid-Borne Enterotoxin Gene (Genotype IS1151-cpe or IS1470-like-cpe) as a Common Cause of Food Poisoning

Journal of Clinical Microbiology ◽

10.1128/jcm.01650-07 ◽

2007 ◽

Vol 46 (1) ◽

pp. 371-373 ◽

Cited By ~ 45

Author(s):

P. Lahti ◽

A. Heikinheimo ◽

T. Johansson ◽

H. Korkeala

Keyword(s):

Clostridium Perfringens ◽

Food Poisoning ◽

Type A ◽

Enterotoxin Gene ◽

Common Cause

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Cloning in Escherichia coli of the Enterotoxin Gene from Clostridium perfringens Type A

Microbiology ◽

10.1099/00221287-135-4-903 ◽

1989 ◽

Vol 135 (4) ◽

pp. 903-909

Author(s):

L. A. Iwanejko ◽

M. N. Routledge ◽

G. S. A. B. Stewart

Keyword(s):

Escherichia Coli ◽

Clostridium Perfringens ◽

Type A ◽

Enterotoxin Gene

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Bicarbonate and amino acids are co-germinants for spores of Clostridium perfringens type A isolates carrying plasmid-borne enterotoxin gene

Food Microbiology ◽

10.1016/j.fm.2017.06.020 ◽

2018 ◽

Vol 69 ◽

pp. 64-71 ◽

Cited By ~ 2

Author(s):

Maryam Alnoman ◽

Pathima Udompijitkul ◽

Saeed Banawas ◽

Mahfuzur R. Sarker

Keyword(s):

Amino Acids ◽

Clostridium Perfringens ◽

Type A ◽

Enterotoxin Gene

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Occurrence and Expression of Toxin Genes in Clostridium Perfringens Isolates from Pigs

Bulletin of the Veterinary Institute in Pulawy ◽

10.2478/v10213-012-0087-0 ◽

2012 ◽

Vol 56 (4) ◽

pp. 495-498 ◽

Cited By ~ 2

Author(s):

Elżbieta Kukier ◽

Magdalena Goldsztejn ◽

Tomasz Grenda ◽

Krzysztof Kwiatek

Keyword(s):

Clostridium Perfringens ◽

Type A ◽

Qualitative Assessment ◽

Toxin Genes ◽

Genes Expression

Abstract Clostridium perfringens isolates were obtained from pigs of five porcine farms in Poland. The presence of C. perfringens was detected in 92% of faeces samples and its number ranged from 1.0 x 101 cfu/g to 1.2 x 107 cfu/g. All the isolates belonged to type A and 48.7% of them contained cpb2 gene. The qualitative assessment of toxin genes expression by type A subtype β2 isolates showed expression of cpa gene in 100% of strains and cpb2 gene in 71% of the analysed strains. The isolate from one-day-old piglets demonstrated also the expression of cpa and cpb2 genes.

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