Clonal diversity of Clostridium perfringens human clinical isolates with various toxin gene profiles based on multilocus sequence typing and alpha-toxin (PLC) typing

Anaerobe ◽  
2021 ◽  
pp. 102473
Author(s):  
Meiji Soe Aung ◽  
Asami Matsuda ◽  
Noriko Urushibara ◽  
Mitsuyo Kawaguchiya ◽  
Nobuhide Ohashi ◽  
...  
Keyword(s):  
Clostridium Perfringens ◽  
Multilocus Sequence Typing ◽  
Clonal Diversity ◽  
Clinical Isolates ◽  
Toxin Gene ◽  
Alpha Toxin ◽  
Gene Profiles

scholarly journals Prevalence and Genetic Diversity of Toxin Genes in Clinical Isolates of Clostridium perfringens: Coexistence of Alpha-Toxin Variant and Binary Enterotoxin Genes (bec/cpile)

Toxins ◽  
2019 ◽  
Vol 11 (6) ◽  
pp. 326 ◽  
Author(s):  
Asami Matsuda ◽  
Meiji Soe Aung ◽  
Noriko Urushibara ◽  
Mitsuyo Kawaguchiya ◽  
Ayako Sumi ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Clostridium Perfringens ◽  
Clinical Isolates ◽  
Nucleotide Sequencing ◽  
Toxin Gene ◽  
Alpha Toxin ◽  
Toxin Genes ◽  
Food Borne ◽  
Putative Origin ◽  
High Level

Clostridium perfringens (C. perfringens) is responsible for food-borne gastroenteritis and other infectious diseases, and toxins produced by this bacterium play a key role in pathogenesis. Although various toxins have been described for C. perfringens isolates from humans and animals, prevalence of individual toxins among clinical isolates has not yet been well explored. In the present study, a total of 798 C. perfringens clinical isolates were investigated for prevalence of eight toxin genes and their genetic diversity by PCR, nucleotide sequencing, and phylogenetic analysis. Besides the alpha-toxin gene (plc) present in all the isolates, the most common toxin gene was cpe (enterotoxin) (34.2%), followed by cpb2 (beta2 toxin) (1.4%), netB (NetB) (0.3%), and bec/cpile (binary enterotoxin BEC/CPILE) (0.1%), while beta-, epsilon-, and iota-toxin genes were not detected. Genetic analysis of toxin genes indicated a high level of conservation of plc, cpe, and netB. In contrast, cpb2 was revealed to be considerably divergent, containing at least two lineages. Alpha-toxin among 46 isolates was classified into ten sequence types, among which common types were distinct from those reported for avian isolates. A single isolate with bec/cpile harbored a plc variant containing an insertion of 834-bp sequence, suggesting its putative origin from chickens.

scholarly journals Quantification of Cell Proliferation and Alpha-Toxin Gene Expression of Clostridium perfringens in the Development of Necrotic Enteritis in Broiler Chickens

2007 ◽  
Vol 73 (21) ◽  
pp. 7110-7113 ◽  
Author(s):  
Weiduo Si ◽  
Joshua Gong ◽  
Yanming Han ◽  
Hai Yu ◽  
John Brennan ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Proliferation ◽  
Regression Model ◽  
Clostridium Perfringens ◽  
Broiler Chickens ◽  
Toxin Gene ◽  
Necrotic Enteritis ◽  
Lesion Score ◽  
Alpha Toxin ◽  
Toxin Gene Expression

ABSTRACT Cell proliferation and alpha-toxin gene expression of Clostridium perfringens in relation to the development of necrotic enteritis (NE) were investigated. Unlike bacitracin-treated chickens, non-bacitracin-treated birds exhibited typical NE symptoms and reduced growth performance. They also demonstrated increased C. perfringens proliferation and alpha-toxin gene expression that were positively correlated and progressed according to the regression model y = b 0 + b 1 X − b 2 X 2. The average C. perfringens count of 5 log10 CFU/g in the ileal digesta appears to be a threshold for developing NE with a lesion score of 2.

scholarly journals Construction of an Alpha Toxin Gene Knockout Mutant of Clostridium perfringens Type A by Use of a Mobile Group II Intron

2005 ◽  
Vol 71 (11) ◽  
pp. 7542-7547 ◽  
Author(s):  
Yue Chen ◽  
Bruce A. McClane ◽  
Derek J. Fisher ◽  
Julian I. Rood ◽  
Phalguni Gupta
Keyword(s):  
Clostridium Perfringens ◽  
Gene Knockout ◽  
Antibiotic Resistance Genes ◽  
Group Ii Intron ◽  
Toxin Gene ◽  
No Production ◽  
Alpha Toxin ◽  
Knockout Mutant ◽  
New Strategy ◽  
Group Ii

ABSTRACT In developing Clostridium perfringens as a safe vaccine vector, the alpha toxin gene (plc) in the bacterial chromosome must be permanently inactivated. Disrupting genes in C. perfringens by traditional mutagenesis methods is very difficult. Therefore, we developed a new strategy using group II intron-based Target-Tron technology to inactivate the plc gene in C. perfringens ATCC 3624. Western blot analysis showed no production of alpha toxin protein in the culture supernatant of the plc mutant. Advantages of this technology, such as site specificity, relatively high frequency of insertion, and introduction of no antibiotic resistance genes into the chromosome, could facilitate construction of other C. perfringens mutants.

scholarly journals Isolation, identification and toxinotyping of Clostridium perfringens isolated from broilers in Pakistan

2021 ◽  
Vol 58 (04) ◽  
pp. 1367-1372
Author(s):  
Zain Ul Abadeen
Keyword(s):  
Clostridium Perfringens ◽  
Causative Agent ◽  
Broiler Chicken ◽  
Clinical Signs ◽  
Type A ◽  
Toxin Gene ◽  
Necrotic Enteritis ◽  
Alpha Toxin ◽  
Enteric Disease ◽  
Isolation Rate

Necrotic enteritis (NE) is one of the important enteric disease in the poultry industry worldwide, caused by C. perfringens type A. This study describes the isolation, identification, and toxinotyping of C. perfringens in necrotic enteritis affected broiler chicken in Pakistan. A total of 430 intestinal samples from dead carcasses and birds suspected of NE outbreak, in and around Faisalabad, Pakistan were collected from 36 broiler farms which yielded 87 alpha toxin gene (cpa) positive C. perfringens type A isolates. The birds having 4-5 weeks of age, clinical signs, and reared in open (conventional) sheds showed higher C. perfringens isolation rate. The study concluded netB negative C. perfringens type A as a causative agent for NE outbreaks in broiler birds in Faisalabad, Pakistan.

scholarly journals Detection of a Group II Intron without an Open Reading Frame in the Alpha-Toxin Gene of Clostridium perfringens Isolated from a Broiler Chicken

2006 ◽  
Vol 189 (5) ◽  
pp. 1633-1640 ◽  
Author(s):  
Menglin Ma ◽  
Kaori Ohtani ◽  
Tohru Shimizu ◽  
Naoaki Misawa
Keyword(s):  
Clostridium Perfringens ◽  
Broiler Chicken ◽  
Group Ii Intron ◽  
Open Reading Frame ◽  
Start Codon ◽  
Northern Hybridization ◽  
Toxin Gene ◽  
Alpha Toxin ◽  
Reading Frame ◽  
Group Ii

ABSTRACTA DNA insertion of 834 bp, designated CPF-G2Im, was identified within the alpha toxin gene (cpa) ofClostridium perfringensstrain CPBC16ML, isolated from a broiler chicken. Sequence analysis of CPF-G2Im indicated that it was integrated 340 nucleotides downstream of the start codon ofcpa. However, the insertion did not abolish the phospholipase C and hemolytic activities of CPBC16ML. To investigate the expression of its alpha toxin, the intact copy ofcpawas cloned into an expression vector and transformed intoEscherichia coliM15 cells. Immunoblotting analysis showed that the protein expressed from the transformant as well as in the culture supernatant ofC. perfringensstrain CPBC16ML had the expected molecular weight detected in reference strains ofC. perfringens. Northern hybridization and reverse transcriptase PCR (RT-PCR) analysis revealed that the entire CPF-G2Im insertion was completely spliced from thecpaprecursor mRNA transcripts. The sequence of the insertion fragment has 95% and 97% identity to two noncoding regions corresponding to sequences that flank a predicted group II RT gene present in the pCPF4969 plasmid ofC. perfringens. However, an RT was not encoded by the CPF-G2Im fragment. Based on the secondary structure prediction analysis, CPF-G2Im revealed typical features of group II introns. The present study shows that CPF-G2Im is capable of splicing in bothC. perfringensandE. coli. To our knowledge, this is the first report that a group II intron without an open reading frame (ORF) is located in thecpaORF ofC. perfringens.

scholarly journals Genotypes and Toxin Gene Profiles of Staphylococcus aureus Clinical Isolates from China

PLoS ONE ◽  
2011 ◽  
Vol 6 (12) ◽  
pp. e28276 ◽  
Author(s):  
Yanping Xie ◽  
Yiping He ◽  
Andrew Gehring ◽  
Yu Hu ◽  
Qiongqiong Li ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Clinical Isolates ◽  
Toxin Gene ◽  
Gene Profiles

Evaluation in broilers of the probiotic properties of Pichia pastoris and a recombinant P. pastoris containing the Clostridium perfringens alpha toxin gene

2012 ◽  
Vol 156 (3-4) ◽  
pp. 448-451 ◽  
Author(s):  
João Rodrigo Gil de los Santos ◽  
Otávio Brod Storch ◽  
Cristina Gevehr Fernandes ◽  
Carlos Gil-Turnes
Keyword(s):  
Pichia Pastoris ◽  
Clostridium Perfringens ◽  
Toxin Gene ◽  
Probiotic Properties ◽  
Alpha Toxin

scholarly journals Recombinant Bacillus subtilis Expressing the Clostridium perfringens Alpha Toxoid Is a Candidate Orally Delivered Vaccine against Necrotic Enteritis

2008 ◽  
Vol 76 (11) ◽  
pp. 5257-5265 ◽  
Author(s):  
Tran H. Hoang ◽  
Huynh A. Hong ◽  
Graeme C. Clark ◽  
Richard W. Titball ◽  
Simon M. Cutting
Keyword(s):  
Bacillus Subtilis ◽  
Clostridium Perfringens ◽  
Vegetative Cell ◽  
Active Agent ◽  
Secretory Iga ◽  
Toxin Gene ◽  
Spore Coat ◽  
Necrotic Enteritis ◽  
Alpha Toxin

ABSTRACT Recombinant Bacillus subtilis endospores have been used to vaccinate against tetanus and anthrax. In this work, we have developed spores that could be used to vaccinate against Clostridium perfringens alpha toxin and that could be used to protect against gas gangrene in humans and necrotic enteritis in poultry. The primary active agent in both cases is alpha toxin. A carboxy-terminal segment of the alpha toxin gene (cpa) fused to the glutathione-S-transferase (GST) gene was cloned in B. subtilis such that the encoded GST-Cpa247-370 polypeptide had been expressed in the following three different ways: expression in the vegetative cell, expression on the surface of the spore coat (fused to the CotB spore coat protein), and a combined approach of spore coat expression coupled with expression in the vegetative cell. Mice immunized orally or nasally with three doses of recombinant spores that carried GST-Cpa247-370 on the spore surface showed the most striking responses. This included seroconversion with anti-Cpa247-370-specific immunoglobulin G (IgG) responses in their sera, a Th2 bias, and secretory IgA responses in saliva, feces, and lung samples. Neutralizing IgG antibodies to alpha toxin were detected using in vitro and in vivo assays, and a toxin challenge established protection. Mice immunized nasally or orally with recombinant spores were protected against a challenge with 12 median lethal doses of alpha toxin. Existing use of spores as competitive exclusion agents in animal feeds supports their use as a potentially economical and heat-stable vaccine for the poultry industry.

The effects of probiotic Bacillus coagulans on the cytotoxicity and expression of alpha toxin gene of Clostridium perfringens type A

Anaerobe ◽  
2019 ◽  
Vol 59 ◽  
pp. 61-67 ◽  
Author(s):  
Amin Kawarizadeh ◽  
Mohammad Tabatabaei ◽  
Saeid Hosseinzadeh ◽  
Mina Farzaneh ◽  
Maryam Pourmontaseri
Keyword(s):  
Clostridium Perfringens ◽  
Bacillus Coagulans ◽  
Type A ◽  
Toxin Gene ◽  
Alpha Toxin

scholarly journals Establishment of a publicly available core genome multilocus sequence typing scheme for Clostridium perfringens

2021 ◽  
Author(s):  
Mostafa Y. Abdel-Glil ◽  
Prasad Thomas ◽  
Jörg Linde ◽  
Keith A. Jolley ◽  
Dag Harmsen ◽  
...  
Keyword(s):  
Clostridium Perfringens ◽  
Multilocus Sequence Typing ◽  
Core Genome ◽  
Epidemiological Studies ◽  
Pathogen Transmission ◽  
Toxin Gene ◽  
Necrotic Enteritis ◽  
Related Factors ◽  
Typing Scheme ◽  
Meat Samples

AbstractClostridium perfringens is a spore-forming anaerobic pathogen responsible for a variety of histotoxic and intestinal infections in humans and animals. High-resolution genotyping aiming to identify bacteria at strain level has become increasingly important in modern microbiology to understand pathogen transmission pathways and to tackle infection sources. This study aimed at establishing a publicly available genome-wide multilocus sequence-typing scheme for C. perfringens. 1,431 highly conserved core genes (1.34 megabases; 50% of the reference genome genes) were indexed for a core-genome-based MLST scheme for C. perfringens. As an example, we applied the scheme to 87 poultry and 73 non-poultry strains (total=160). The genotyping results of the 160 genomes were congruent in terms of resolution and tree topology between allele-based and single-nucleotide-polymorphism-based core-genome typing. For the analysis of poultry strains of C. perfringens concerning the country of isolation, NetB-toxin gene carriage and clinical disease, we used 60 allelic differences as a clustering threshold. The results showed that poultry strains from a single country formed a cluster (n=17 clusters including 46 strains). Two clusters included six strains from four different countries. These strains were netB-positive, as were seven strains from Denmark and two strains from Finland, possibly indicating common sources of netB-positive strains. In terms of clinical presentation, different clusters of strains were associated with cases of suspected necrotic enteritis. Strains from sick birds grouped with strains from healthy birds or meat samples showing that potentially virulent strains are widespread and that host-related factors contribute significantly to NE. In summary, a publicly available scheme and an allele nomenclature database for genomic typing of C. perfringens has been established and can be used for broad-based and standardised epidemiological studies.

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