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.

Regulation of toxin gene expression in Clostridium perfringens

2015 ◽  
Vol 166 (4) ◽  
pp. 280-289 ◽  
Author(s):  
Kaori Ohtani ◽  
Tohru Shimizu
Keyword(s):  
Gene Expression ◽  
Clostridium Perfringens ◽  
Toxin Gene ◽  
Toxin Gene Expression

scholarly journals Immunization of Broiler Chickens against Clostridium perfringens-Induced Necrotic Enteritis

2007 ◽  
Vol 14 (9) ◽  
pp. 1070-1077 ◽  
Author(s):  
R. R. Kulkarni ◽  
V. R. Parreira ◽  
S. Sharif ◽  
J. F. Prescott
Keyword(s):  
Clostridium Perfringens ◽  
Broiler Chickens ◽  
Hypothetical Protein ◽  
Oral Infection ◽  
Secreted Proteins ◽  
Enzyme Linked Immunosorbent Assay ◽  
Necrotic Enteritis ◽  
Alpha Toxin ◽  
Genes Encoding ◽  
Antibody Titers

ABSTRACT Necrotic enteritis (NE) in broiler chickens is caused by Clostridium perfringens. Currently, no vaccine against NE is available and immunity to NE is not well characterized. Our previous studies showed that immunity to NE followed oral infection by virulent rather than avirulent C. perfringens strains and identified immunogenic secreted proteins apparently uniquely produced by virulent C. perfringens isolates. These proteins were alpha-toxin, glyceraldehyde-3-phosphate dehydrogenase, pyruvate:ferredoxin oxidoreductase (PFOR), fructose 1,6-biphosphate aldolase, and a hypothetical protein (HP). The current study investigated the role of each of these proteins in conferring protection to broiler chickens against oral infection challenges of different severities with virulent C. perfringens. The genes encoding these proteins were cloned and purified as histidine-tagged recombinant proteins from Escherichia coli and were used to immunize broiler chickens intramuscularly. Serum and intestinal antibody responses were assessed by enzyme-linked immunosorbent assay. All proteins significantly protected broiler chickens against a relatively mild challenge. In addition, immunization with alpha-toxin, HP, and PFOR also offered significant protection against a more severe challenge. When the birds were primed with alpha-toxoid and boosted with active toxin, birds immunized with alpha-toxin were provided with the greatest protection against a severe challenge. The serum and intestinal washings from protected birds had high antigen-specific antibody titers. Thus, we conclude that there are certain secreted proteins, in addition to alpha-toxin, that are involved in immunity to NE in broiler chickens.

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 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.

scholarly journals Evaluation of Candidate Reference Genes Stability for Gene Expression Analysis by Reverse Transcription QPCR in Clostridium Perfringens

2021 ◽  
Author(s):  
Michele Williams ◽  
Mostafa Ghanem
Keyword(s):  
Gene Expression ◽  
Clostridium Perfringens ◽  
Reference Gene ◽  
Reverse Transcription ◽  
Reference Genes ◽  
Quantitative Polymerase Chain Reaction ◽  
Toxin Gene ◽  
Rrna Gene ◽  
Experimental Conditions ◽  
Toxin Gene Expression

Abstract Identification of stable reference genes for normalization purposes is necessary for obtaining reliable and accurate results of reverse transcription quantitative polymerase chain reaction (RT-qPCR) analyses. To our knowledge, no reference gene(s) have been validated for this purpose in Clostridium perfringens. In this study, the expression profile of ten candidate reference genes from three strains of C. perfringens were assessed for stability under various experimental conditions using geNorm in qbase+. These stability rankings were then compared to stability assessments evaluated by BestKeeper, NormFinder, delta Ct, and RefFinder algorithms. When comparing all the analyses; gyrA, ftsZ, and recA were identified within the most stable genes under the different experimental conditions and were further tested as a set of reference genes for normalization of alpha toxin gene expression over a 22-hour period. Depending on the condition, rpoA and rho might also be suitable to include as part of the reference set. Although commonly used for the purpose of normalizing RT-qPCR data, the 16S rRNA gene (rrs) was found to be an unsuitable gene to be used as a reference. This work provides a framework for the selection of a suitable stable reference gene set for data normalization of C. perfringens gene expression.

scholarly journals The effects of Pediococcus acidilactici and Saccharomyces cerevisiae on broiler chickens challenged with Clostridium perfringens induced subclinical necrotic enteritis

2021 ◽  
Vol 91 (4) ◽  
pp. 389-397
Author(s):  
Rachid Merati ◽  
◽  
Ali Alaa Abdel-Fattah Mohamed ◽  
Zahra Berrama ◽  
Hebib Aggad ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Growth Performance ◽  
Clostridium Perfringens ◽  
Broiler Chickens ◽  
Body Weight Gain ◽  
Control Group ◽  
Necrotic Enteritis ◽  
Pediococcus Acidilactici ◽  
Lesion Score ◽  
Feed Conversion

The objective of this study was to evaluate the effects of Pediococcus acidilactici (P. acidilactici) and Saccharomyces cerevisiae (S. cerevisiae) on the growth performance, intestinal lesions and ileal Clostridium perfringens (C. perfringens) count of broiler chickens challenged with C. perfringens induced sub-clinical necrotic enteritis (NE). A total of 150 broiler chicks, allocated into five treatment groups (6 replicates of 5 chicks/cage), were reared in cages for 29 days: T0 (control group, not infected, not supplemented); T1 (infected with C. perfringens, not supplemented); T2 (infected with C. perfringens and coccidia, not supplemented); T3 (infected with C. perfringens and coccidia, supplemented with P. acidilactici); T4 (infected with C. perfringens and coccidia, supplemented with S. cerevisiae). The parameters analyzed were: body weight gain, feed intake, feed conversion ratio, mortality, intestinal lesion score and ileal C. perfringens enumeration by quantitative real-time PCR. The challenge resulted in impairment of growth performance, increased lesion score (≤ 2) and overgrowth of the C. perfringens population. However, the dietary inclusion of P. acidilactici or S. cerevisiae caused a significant improvement in feed conversion, net reduction of gut lesions, as well as a decrease in the intestinal C. perfringens population. In conclusion, these results suggest that dietary supplementation with probiotics (P. acidilactici or S. cerevisiae) could be beneficial to alleviate the negative effects of subclinical NE in broiler chickens.

scholarly journals Supplementing Garlic Nanohydrogel Optimized Growth, Gastrointestinal Integrity and Economics and Ameliorated Necrotic Enteritis in Broiler Chickens Using a Clostridium perfringens Challenge Model

Animals ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 2027
Author(s):  
Doaa Ibrahim ◽  
Tamer Ahmed Ismail ◽  
Eman Khalifa ◽  
Shaimaa A. Abd El-Kader ◽  
Dalia Ibrahim Mohamed ◽  
...  
Keyword(s):  
Growth Performance ◽  
Clostridium Perfringens ◽  
Broiler Chickens ◽  
Body Weight Gain ◽  
Economic Benefits ◽  
Production Costs ◽  
Broiler Chicks ◽  
Necrotic Enteritis ◽  
Feed Conversion ◽  
Intestinal Lesion

Necrotic enteritis (NE) caused by Clostridium perfringens (C. perfringens) results in impaired bird growth performance and increased production costs. Nanotechnology application in the poultry industry to control NE outbreaks is still not completely clarified. Therefore, the efficacy of dietary garlic nano-hydrogel (G-NHG) on broilers growth performance, intestinal integrity, economic returns and its potency to alleviate C. perfringens levels using NE challenge model were addressed. A total of 1200 male broiler chicks (Ross 308) were assigned into six groups; four supplemented with 100, 200, 300 or 400 mg of G-NHG/kg diet and co-challenged with C. perfringens at 21, 22 and 23 d of age and two control groups fed basal diet with or without C. perfringens challenge. Over the total growing period, the 400 mg/kg G-NHG group had the most improved body weight gain and feed conversion efficiency regardless of challenge. Parallel with these results, the mRNA expression of genes encoding digestive enzymes (alpha 2A amylase (AMY2A), pancreatic lipase (PNLIP) and cholecystokinin (CCK)) and intestinal barriers (junctional adhesion molecule-2 (JAM-2), occludin and mucin-2 (Muc-2)) were increased in groups fed G-NHG at higher levels to be nearly similar to those in the unchallenged group. At 14 d post challenge, real-time PCR results revealed that inclusion of G-NHG led to a dose-dependently decrease in the C. perfringens population, thereby decreasing the birds’ intestinal lesion score and mortality rates. Using 400 mg/kg of G-NHG remarkably ameliorated the adverse effects of NE caused by C. perfringens challenge, which contributed to better growth performance of challenged birds with rational economic benefits.

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