Molecular detection and characterization of cpb2 gene in Clostridium perfringens isolates from healthy and diseased chickens

Tolooe, Ali; Shojadoost, Bahram; Peighambari, Seyed Mostafa

doi:10.1590/s1678-91992011000100008

Cited by 11 publications

(6 citation statements)

References 19 publications

(32 reference statements)

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“…Throughout the last decade, several epidemiological studies showed wide distribution of beta 2 (β 2 ) toxigenic C. perfringens strains among human and other animal species [49] but its exact role in pathogenesis would still to be further elucidated [50]. In this study, it was discovered in both diseased and healthy birds in percentages of 75% and 71.4%.…”

Section: Discussionmentioning

confidence: 61%

“…Differently, the difference of nucleotide sequences at positions 6, 10, 12, 20 and 198 of two Iranian C. perfringens isolates was recorded [49] with 99% similarity to each other and 73 % identity with the cpb 2 sequences of C. perfringens strains. An absence of β2 toxin expression where almost half of the non-porcine consensus cpb 2 genes (44.4%) carried a frameshift mutation was also, reported [56].…”

Section: Discussionmentioning

confidence: 86%

Toxin Genotyping of <i>C. perfringens</i> Isolated from Broiler Cases of Necrotic Enteritis

Ibrahim¹

2017

AVS

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Clostridium perfringens organisms have an economic concern in poultry production. The purpose of this study was to investigate Net B and β 2 C. perfringens positive isolates in broiler farms and their clinic-pathological effects in broiler chicks. A bacteriological examination of C. perfringens was carried upon 92 Necrotic enteritis (NE) diseased cases and 55 apparently healthy broilers of different ages in Egypt. C. perfringens type A was only recovered (49.7%). cpa gene was detected in 100% of samples with PCR technique. NE diseased cases exhibited both Net B (87.5%) and cpb 2 (75%) toxin genes. Experimentally, an intra-gut induction of Net B and β 2 C. perfringens toxins were evaluated in chicken models. The hematological studies revealed hemolytic anemia 5 days post infection (p.i) in Net B and β 2 inoculated groups (G1&G2). Leucogram revealed neutrophilia and lymphopenia 5 days p.i. A significant increase in ALT, AST, uric acid and creatinine serum levels were recorded in the infected groups at 5 th and 12 th day p.i. DNA Sequencing for net B gene revealed an amino acid replacement from glutamate into arginine at codon 379 with silent mutation was also detected at nucleotide 1134. Sequencing of both toxin genes were recorded in the gene bank for the first time in Egypt. This study pointed out that C. perfringens Net B toxin, is a new key virulent factor for the development of NE. Further studies of Net B toxiod for vaccine production could minimize the clostridial problems in broiler farms.

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“…Nonetheless, some of these species are associated with very serious diseases (28). The pathogenicity of this bacterium is associated with the production of extracellular toxins produced by some of its strains, such as beta2 toxin (29). The incidence of the disease may have been exacerbated by the use of wheat and barley as basal ingredients in European poultry diets.…”

Section: Discussionmentioning

confidence: 99%

Incidence of Clostridium perfringens in Intestinal Contents of Domestic Livestock Detected by PCR

Arshi¹,

Nikkhah²,

Kabiri³

et al. 2017

IJAR

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“…The cpb2 gene carried by strain EHE-NE18 is identical to the atypical version. Another recent study found that two C. perfringens isolates from diseased chickens also carried the atypical cpb2 gene ( 55 ). Preliminary analysis suggests that many netB -positive C. perfringens strains have the atypical cpb2 gene, including the Belgian strain 56.…”

Section: Discussionmentioning

confidence: 99%

Necrotic Enteritis-Derived Clostridium perfringens Strain with Three Closely Related Independently Conjugative Toxin and Antibiotic Resistance Plasmids

Bannam

Yan

Harrison

et al. 2011

mBio

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The pathogenesis of avian necrotic enteritis involves NetB, a pore-forming toxin produced by virulent avian isolates of Clostridium perfringens type A. To determine the location and mobility of the netB structural gene, we examined a derivative of the tetracycline-resistant necrotic enteritis strain EHE-NE18, in which netB was insertionally inactivated by the chloramphenicol and thiamphenicol resistance gene catP. Both tetracycline and thiamphenicol resistance could be transferred either together or separately to a recipient strain in plate matings. The separate transconjugants could act as donors in subsequent matings, which demonstrated that the tetracycline resistance determinant and the netB gene were present on different conjugative elements. Large plasmids were isolated from the transconjugants and analyzed by high-throughput sequencing. Analysis of the resultant data indicated that there were actually three large conjugative plasmids present in the original strain, each with its own toxin or antibiotic resistance locus. Each plasmid contained a highly conserved 40-kb region that included plasmid replication and transfer regions that were closely related to the 47-kb conjugative tetracycline resistance plasmid pCW3 from C. perfringens. The plasmids were as follows: (i) a conjugative 49-kb tetracycline resistance plasmid that was very similar to pCW3, (ii) a conjugative 82-kb plasmid that contained the netB gene and other potential virulence genes, and (iii) a 70-kb plasmid that carried the cpb2 gene, which encodes a different pore-forming toxin, beta2 toxin.

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“…These findings were in correlation with Lyhs et al (2013) reported that the netB was identified in 6.6 percent from C. perfringens strains which were isolated from NE cases in turkeys. Tolooe et al (2011) who reported, 52.8 percent prevalence of netB from diseased birds and none from healthy birds. Ezatkhah and associates (2016) reported netB gene for the first time at a low incidence (7.77%) in chickens with NE in organic broiler farms and detected 17.98 percent prevalence by PCR.…”

Section: Detection Of Virulent Gene Netb Of C Perfringensmentioning

confidence: 97%

Toxinotyping of Clostridium perfringens in Poultry from Necrotic Enteritis Cases

Shelke¹

2020

JAR

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Clostridium perfringens is well known causative agent of necrotic enteritis in poultry and is mainly caused by Type A toxin. NetB toxin is found to be one of the newly emerging virulent toxin gene which is also responsible for necrotic enteritis. The present study was carried out to characterize and to type the different toxins associated with C. perfringens in NE cases of poultry. For the present study total 125 samples were collected from poultry birds, out of which 50 samples were of intestinal content from diseased birds, 50 cloacal swabs and 25 intestinal content from healthy birds. These samples were further processed for isolation, identification, and toxinotyping of Clostridium perfringens isolates. Onisolation of C. perfringens on blood agar total 43 isolates were found positive showing a pattern of double hemolysis on blood agar. The positive isolates of C. perfringens were further confirmed by using 16S rRNA species specific PCR. After confirmation isolates were processed for toxinotyping mainly targeting cpa, cpb and cpb2 toxins by using multiplex PCR. On toxinotyping it was found that NE in poultry birds were mainly caused by C. perfringens type A. On virulent gene detection of netB toxin, total 4 isolates were found positive for netB toxin. This study pointed out that C. perfringens type A is responsible for development of NE in poultry along with net B toxin which is a new key virulent factor. Further studies of netB toxoid and C. perfringens type A for vaccine production could minimize the clostridial problems in broiler farms.

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Molecular detection and characterization of cpb2 gene in Clostridium perfringens isolates from healthy and diseased chickens

Cited by 11 publications

References 19 publications

Toxin Genotyping of <i>C. perfringens</i> Isolated from Broiler Cases of Necrotic Enteritis

Incidence of Clostridium perfringens in Intestinal Contents of Domestic Livestock Detected by PCR

Necrotic Enteritis-Derived Clostridium perfringens Strain with Three Closely Related Independently Conjugative Toxin and Antibiotic Resistance Plasmids

Toxinotyping of Clostridium perfringens in Poultry from Necrotic Enteritis Cases

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Molecular detection and characterization of cpb2 gene in Clostridium perfringens isolates from healthy and diseased chickens

Cited by 11 publications

References 19 publications

Toxin Genotyping of &lt;i&gt;C. perfringens&lt;/i&gt; Isolated from Broiler Cases of Necrotic Enteritis

Incidence of Clostridium perfringens in Intestinal Contents of Domestic Livestock Detected by PCR

Necrotic Enteritis-Derived Clostridium perfringens Strain with Three Closely Related Independently Conjugative Toxin and Antibiotic Resistance Plasmids

Toxinotyping of Clostridium perfringens in Poultry from Necrotic Enteritis Cases

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Toxin Genotyping of <i>C. perfringens</i> Isolated from Broiler Cases of Necrotic Enteritis