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 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.

Clostridium perfringens Type A from Broiler Chicken with Necrotic Enteritis

2008 ◽  
Vol 7 (6) ◽  
pp. 601-609 ◽  
Author(s):  
Arunava Das ◽  
Yahya Mazumder ◽  
Biman Kumar Dutta ◽  
Bibek Ranjan Shome ◽  
Komal Molla Bujarbarua ◽  
...  
Keyword(s):  
Clostridium Perfringens ◽  
Broiler Chicken ◽  
Type A ◽  
Necrotic Enteritis

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 Antimicrobial Susceptibility and Association with Toxin Determinants in Clostridium perfringens Isolates from Chickens

2020 ◽  
Vol 8 (11) ◽  
pp. 1825
Author(s):  
Bai Wei ◽  
Se-Yeoun Cha ◽  
Jun-Feng Zhang ◽  
Ke Shang ◽  
Hae-Chul Park ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Clostridium Perfringens ◽  
Clinical Signs ◽  
Toxin Gene ◽  
Necrotic Enteritis ◽  
Growth Promoters ◽  
Complete Understanding ◽  
Toxin Genes ◽  
Antimicrobial Growth Promoters ◽  
Selection Of

The aim of the present study was to investigate variation in antimicrobial resistance in Clostridium perfringens (C. perfringens) isolated from chickens after withdrawal of antimicrobial growth promoters (AGPs); and to investigate the correlation between the presence of toxin genes (cpb2, netB, and tpeL) and antimicrobial resistance. Altogether, 162 isolates of C. perfringens were obtained from chickens displaying clinical signs of necrotic enteritis (n = 65) and from healthy chickens (n = 97) in Korea during 2010–2016. Compared to before AGP withdrawal, increased antimicrobial resistance or MIC50/MIC90 value was observed for nine antimicrobials including penicillin, tetracycline, tylosin, erythromycin, florfenicol, enrofloxacin, monensin, salinomycin, and maduramycin. Significantly (p < 0.05) higher resistance to gentamicin, clindamycin, and virginiamycin was found in isolates from chickens with necrotic enteritis compared to those from healthy chickens. tpeL gene was not detected in C. perfringens isolates from healthy chickens. A correlation between toxin gene prevalence and antibiotic resistance was found in the C. perfringens isolates. Because the usage of antimicrobials may contribute to the selection of both resistance and toxin genes, these can potentially make it challenging to control antimicrobial resistance in pathogenic colonies. Therefore, a more complete understanding of the interplay between resistance and virulence genes is required.

scholarly journals Genetic Diversity of Clostridium perfringens Strains Isolated from Broiler Chickens Revealed by PFGE Analysis in China and Pakistan

2021 ◽  
Vol 41 (01) ◽  
pp. 85-91
Author(s):  
Muhammad Umar Zafar Khan
Keyword(s):  
Clostridium Perfringens ◽  
Broiler Chickens ◽  
Broiler Chicken ◽  
Control Strategies ◽  
Type A ◽  
Necrotic Enteritis ◽  
Pfge Profile ◽  
Pfge Analysis ◽  
Lack Of Information ◽  
Beta 2

Clostridium perfringens (C. perfringens) is widely distributed in broiler chickens causing clinical and subclinical enteritis and is especially known for causing necrotic enteritis (NE). There are numerous reports of NE outbreaks in Pakistan as well as China but there is a lack of information related to PFGE profile from both the countries. To close this gap, we designed this study and obtained samples from broiler chicken farms located in 3 different regions of Pakistan and 4 different regions of China. A total of 79 fecal swabs (Pakistan=29; China=50) were collected and grown on FTA media. Further, isolates were grown on TSE agar and black colonies were selected for DNA extraction. All 79 isolates were tested for toxin profiles by PCR (α-gene; beta-2; netB gene) and PFGE profiling (pulsotypes analysis). Toxinotyping results revealed that all the isolates (n=50) from China were type A (α-toxin positive) while 23 and 6 isolates (n=29) from Pakistan were type A (α-toxin positive) and type G (α-toxin, NetB positive), respectively. Toxinotyping revealed α-toxin is highly prevalent in both the countries while from Pakistani isolates, NetB toxin was also detected. PFGE discriminated 79 isolates into 45 different PFGE patterns (pulsotypes). The analysis further showed different pulsotypes originating from China and Pakistan and isolates were subtyped by SmaI. The results showed high genetic polymorphism in C. perfringens even within the same strain. These preliminary findings of genetic variations will further help to design control strategies

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 Isolation and Genotyping Study of Clostridium Perfringens From Broiler Farms Infected with Necrotic Enteritis in Sulaimania Province

2019 ◽  
Vol 12 (2) ◽  
Keyword(s):  
Sequence Analysis ◽  
Dna Sequence ◽  
Clostridium Perfringens ◽  
Causative Agent ◽  
Type A ◽  
Dna Sequence Analysis ◽  
Necrotic Enteritis ◽  
Intestinal Contents ◽  
Lymphoid Nodules ◽  
Mucosal Scraping

The study is conducting to isolate and toxinotype the suspected cases of Clostridium perfringens infections of broiler farms in Sulaimania province. A total of 108 samples were collected from intestinal contents, mucosal scraping, and hemorrhagic lymphoid nodules from suspected cases of necrotic enteritis in broilers. The result of isolated and identified bacteria were revealed that 63 (58%) out of 108 samples were positive for C. perfringens. The results revealed that the isolates were only positive for alpha and beta2 toxin genes. Phylogenetic and DNA sequence analysis of cpa and cpb2 gene showed that cpa genes were highly identical to isolates from broiler in Iran, poultry stool and broiler in Brazil, and blue calves in Belgium. While cpb2 gene is closely related to the isolates of broiler in Iran, India and isolates of goat in Pakistan. The results indicated that the causative agent of necrotic enteritis in broiler farms in the region was mainly due to C. perfringens type A infection

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