Genetic diversity of Clostridium perfringens isolated from healthy broiler chickens at a commercial farm

Chalmers, Gabhan; Martin, S W; Hunter, Douglas B.; Prescott, John F.; Weber, L J; Boerlin, Patrick

doi:10.1016/j.vetmic.2007.08.008

Cited by 58 publications

(48 citation statements)

References 31 publications

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“…In this paper, net B toxin of C. perfringens was studied and detected in NE diseased broilers in a percentage of (46.7%) but didn't found in the isolates from healthy birds. These results were in accordance with a study [47] in which they stated that net B gene was only detected in Candian isolates that were associated with NE outbreaks but it wasn't found in isolates from healthy birds. In addition, net B gene was found in 77.8%, 74.4% and 70% in chickens derived NE C. perfringens strains [9,18,48].…”

Section: Discussionsupporting

confidence: 92%

“…In this study, it was discovered in both diseased and healthy birds in percentages of 75% and 71.4%. Similarl studies [51,47,36] detected cpb 2 toxin gene in 75%, 74.2% and 62.6% of C. perfringens type A isolates in NE affected chickens. C. perfringens isolates were not capable of causing disease without net B gene especially it is linked with the health condition of the bird while a weak or no relationship between β 2 toxin and NE disease in birds [8,46].…”

Section: Discussionsupporting

confidence: 65%

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

Ibrahim¹

2017

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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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Section: Discussionsupporting

confidence: 92%

Section: Discussionsupporting

confidence: 65%

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

Ibrahim¹

2017

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“…However, there is a growing body of evidence that suggests that the beta-2 toxin is not a critical factor of C. perfringens infection in this particular case of poultry (4,6,30). The recently discovered netB toxin gene, however, was found exclusively in isolates which were all associated with NE outbreaks and was never detected in healthy control samples from processor 2.…”

Section: Discussionmentioning

confidence: 91%

“…These methods include, among others, pulsed-field gel electrophoresis (4,7,23), multilocus variable number of tandem repeats analysis (5,25), and multilocus sequence typing (MLST) (16). MLST is a procedure for characterizing bacteria by using the sequences of several "house-keeping" genes (20).…”

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Multilocus Sequence Typing Analysis ofClostridium perfringensIsolates from Necrotic Enteritis Outbreaks in Broiler Chicken Populations

et al. 2008

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Clostridium perfringens is an important pathogen of animals and humans and is the causative agent of necrotic enteritis (NE) in poultry. This study focuses on the typing of intestinal C. perfringens isolates (n ‫؍‬ 61) from outbreaks of NE collected from several areas of Southern Ontario, using a recently developed multilocus sequence typing (MLST) technique. For comparison, C. perfringens isolates from healthy birds were also obtained and typed. An additional locus, the pfoS locus, was included in our analysis, in an attempt to increase the discriminatory ability of the method previously published. Birds were collected from two major poultry processors in Canada, and isolates from processor 2 formed a distinct MLST cluster. Isolates from healthy birds also collected from the outbreak flocks clustered together with isolates from the birds with NE. Although isolates from eight outbreaks clustered together, MLST types were also occasionally different between outbreaks. Strong linkage disequilibrium was observed between loci, suggesting a clonal C. perfringens population structure. Detection assays for toxin genes cpb2 (beta-2 toxin), tpeL, and the newly described netB (NetB toxin) were also performed. netB was almost always found in outbreak isolates, whereas cpb2 was found exclusively in healthy bird isolates. The toxin gene tpeL, which has not been previously identified in C. perfringens type A strains, was also found, but only in the presence of netB. Resistance to bacitracin was found in 34% of isolates from antimicrobial agent-free birds and in 100% of isolates from conventionally raised birds.

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“…Several of the 54 C. perfringens poultry isolates examined in the current study had been typed by disparate methods, including MLST, PFGE, and multilocus variable number of tandem repeats analysis (MLVA), in previous studies (14,23,24). To assess the genetic relatedness of the isolates using a common method, all were typed by PFGE, with the exception of one strain (JP77) that was not typeable (see Fig.…”

Section: Resultsmentioning

confidence: 99%

Identification of Accessory Genome Regions in Poultry Clostridium perfringens Isolates Carrying the netB Plasmid

Lepp

Gong

Songer

et al. 2013

Journal of Bacteriology

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c Necrotic enteritis (NE) is an economically important disease of poultry caused by certain Clostridium perfringens type A strains. NE pathogenesis involves the NetB toxin, which is encoded on a large conjugative plasmid within a 42-kb pathogenicity locus. Recent multilocus sequence type (MLST) studies have identified two predominant NE-associated clonal groups, suggesting that host genes are also involved in NE pathogenesis. We used microarray comparative genomic hybridization (CGH) to assess the gene content of 54 poultry isolates from birds that were healthy or that suffered from NE. A total of 400 genes were variably present among the poultry isolates and nine nonpoultry strains, many of which had putative functions related to nutrient uptake and metabolism and cell wall and capsule biosynthesis. The variable genes were organized into 142 genomic regions, 49 of which contained genes significantly associated with netB-positive isolates. These regions included three previously identified NE-associated loci as well as several apparent fitness-related loci, such as a carbohydrate ABC transporter, a ferric-iron siderophore uptake system, and an adhesion locus. Additional loci were related to plasmid maintenance. Cluster analysis of the CGH data grouped all of the netB-positive poultry isolates into two major groups, separated according to two prevalent clonal groups based on MLST analysis. This study identifies chromosomal loci associated with netB-positive poultry strains, suggesting that the chromosomal background can confer a selective advantage to NE-causing strains, possibly through mechanisms involving iron acquisition, carbohydrate metabolism, and plasmid maintenance. C lostridium perfringens is an important Gram-positive anaerobic pathogen of humans and animals and is found ubiquitously in soil and in the gastrointestinal tract of vertebrates. It causes a number of histotoxic and enterotoxemic diseases, including necrotic enteritis (NE), an economically important disease of poultry. NE is characterized by necrotic lesions in the small intestine and can occur in an acute form, which is often responsible for high flock mortality, and a subclinical form, which results in production losses (1). A novel toxin, NetB, is present in the majority of disease-associated isolates and plays a critical role in NE pathogenesis (2, 3).As a species, C. perfringens produces an array of extracellular toxins, four of which (alpha, beta, epsilon, and iota) form the basis for a toxin-typing scheme (4). Several of these toxins, including beta2-toxin, C. perfringens enterotoxin (CPE) (in non-food-borne strains), and all of the typing toxins except for alpha-toxin, are encoded on a conserved family of large plasmids related to the pCW3 tetracycline resistance plasmid (5-7). These plasmids share a conserved core region that includes the transfer of the clostridial plasmid (tcp) locus required for conjugation (6,8,9). The gene encoding NetB was recently localized to a 42-kb pathogenicity locus, NELoc-1, which resides on an ϳ85-kb p...

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Genetic diversity of Clostridium perfringens isolated from healthy broiler chickens at a commercial farm

Cited by 58 publications

References 31 publications

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

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

Multilocus Sequence Typing Analysis ofClostridium perfringensIsolates from Necrotic Enteritis Outbreaks in Broiler Chicken Populations

Identification of Accessory Genome Regions in Poultry Clostridium perfringens Isolates Carrying the netB Plasmid

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Genetic diversity of Clostridium perfringens isolated from healthy broiler chickens at a commercial farm

Cited by 58 publications

References 31 publications

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

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

Multilocus Sequence Typing Analysis ofClostridium perfringensIsolates from Necrotic Enteritis Outbreaks in Broiler Chicken Populations

Identification of Accessory Genome Regions in Poultry Clostridium perfringens Isolates Carrying the netB Plasmid

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

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