Role of Clostridium perfringens Toxins in Necrotic Enteritis in Poultry

ABSTRACT Clostridium perfringens encodes at least two different quorum sensing (QS) systems, the Agr-like and LuxS, and recent studies have highlighted their importance in the regulation of toxin production and virulence. The role of QS in the pathogenesis of necrotic enteritis (NE) in poultry and the regulation of NetB, the key toxin involved, has not yet been investigated. We have generated isogenic agrB-null and complemented strains from parent strain CP1 and demonstrated that the virulence of the agrB-null mutant was strongly attenuated in a chicken NE model system and restored by complementation. The production of NetB, a key NE-associated toxin, was dramatically reduced in the agrB mutant at both the transcriptional and protein levels, though not in a luxS mutant. Transwell assays confirmed that the Agr-like QS system controls NetB production through a diffusible signal. Global gene expression analysis of the agrB mutant identified additional genes modulated by Agr-like QS, including operons related to phospholipid metabolism and adherence, which may also play a role in NE pathogenesis. This study provides the first evidence that the Agr-like QS system is critical for NE pathogenesis and identifies a number of Agr-regulated genes, most notably netB, that are potentially involved in mediating its effects. The Agr-like QS system thus may serve as a target for developing novel interventions to prevent NE in chickens.

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ROLE OF CLOSTRIDIUM PERFRINGENS IN PATHOGENICITY OF SOME DOMESTIC ANIMALS

JOURNAL OF ADVANCES IN AGRICULTURE ◽

10.24297/jaa.v7i3.6325 ◽

2017 ◽

Vol 7 (3) ◽

pp. 1117-1121 ◽

Cited By ~ 1

Author(s):

Mohammad Reza Mohammadabadi

Keyword(s):

Clostridium Perfringens ◽

Gastrointestinal Disease ◽

Food Poisoning ◽

Small Ruminants ◽

Necrotic Enteritis ◽

Economic Stability ◽

Native Breed ◽

The World ◽

The Tropics

Clostridium perfringens, is an anaerobic, gram-positive, pathogenic and spore-forming bacillus and broadly gave out in our territory. This bacterium has spore formation capability and creating gangrene and gastrointestinal disease, for example food poisoning and necrotic enteritis in human, whilst in other animals, gastrointestinal and enterotoxemic diseases more happening. Prevalence of necrotic enteritis, created by C. perfringens, has been often stated in sheep, chickens and ostrich throughout the world. The most critical problem for epidemiological investigations and vaccines improvement is accurate recognition of C. perfringens variants. Moreover, Small ruminants, especially native breed types, play an important role to the livelihoods of a considerable part of human population in the tropics from socio-economic aspects. Therefore, integrated attempt in terms of management and genetic improvement to enhance production is of crucial importance. Poultry provide humans with companionship, food and fiber in the form of eggs, meat and feathers. Many people love to raise and show chickens and other poultry species at fairs and other poultry shows. Others just love to raise them for backyard pets and for fresh eggs every day. In the last few years, ostrich farming has progressed dramatically and the world ostrich industry has achieved some economic stability. There is considerable scope for improvement in the areas of artificial incubation, chick nutrition, environmental requirements and selective breeding. Hence, the aim of this paper was to study role of Clostridium perfringens in pathogenicity of sheep, broilers and Ostrich. In conclusion, recognition of toxins producing by C. perfringens is very momentous because their toxin types are related to particular gastric and intestinal animal sickness and PCR has become an essential research and diagnostic tool, being a powerful technique with a vast and increasing range of applications. Hence, it is better that animal breeders identify different types of C. perfringens using PCR technique to prevent the damage caused by this bacterium.

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Developing an experimental necrotic enteritis model in turkeys - the impact of Clostridium perfringens, Eimeria meleagrimitis and host age on frequency of severe intestinal lesions

10.21203/rs.2.12449/v2 ◽

2019 ◽

Author(s):

Simon P. Hardy ◽

Sylvie L. Benestad ◽

Inger Sofie Hamnes ◽

Torfinn Moldal ◽

Bruce David ◽

...

Keyword(s):

Clostridium Perfringens ◽

Scoring System ◽

In Vivo Model ◽

Necrotic Enteritis ◽

Intestinal Lesions ◽

Gross Lesions ◽

Successful Establishment ◽

The Impact

Abstract Background: Necrotic enteritis is a significant problem to the poultry industry globally and, in Norway up to 30 % of Norwegian turkey grow-outs can be affected. However, despite an awareness that differences exist between necrotic enteritis in chickens and turkeys, little information exists concerning the pathogenesis, immunity, microbiota or experimental reproduction of necrotic enteritis in turkeys. In particular, it is important to determine the appearance of the gross lesions, the age dependency of the disease and the role of netB toxin of Clostridium perfringens . To this end, we report our findings in developing an in vivo experimental model of necrotic enteritis in turkeys. Results: A four tier (0-3) scoring system with clearly defined degrees of severity of macroscopic intestinal lesions was developed, based on 2312 photographic images of opened intestines from 810 B.U.T. 10 or B.U.T. Premium turkeys examined in nine experiments. Loss of macroscopically recognizable villi in the anterior small intestine was established as the defining lesion qualifying for a score 3 (severe intestinal lesions). The developed scoring system was used to identify important factors in promoting high frequencies of turkeys with severe lesions: a combined Eimeria meleagrimitis and Clostridium perfringens challenge, challenge at five rather than three weeks of age, the use of an Eimeria meleagrimitis dose level of at least 5 000 oocysts per bird and finally, examination of the intestines of 5-week-old turkeys at 125 to 145 hours after Eimeria meleagrimitis inoculation. Numbers of oocysts excreted were not influenced by Clostridium perfringens inoculation or turkey age. Among three different lesion score outcomes tested, frequency of severe lesions proved superior in discriminating between impact of four combinations of Clostridium perfringens inoculation and turkey age at challenge. Conclusions: This study provides details for the successful establishment of an in vivo model of necrotic enteritis in turkeys.

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A novel PilR/PilS two-component system regulates NE pilus production in Clostridium perfringens

Journal of Bacteriology ◽

10.1128/jb.00096-21 ◽

2021 ◽

Author(s):

Yuanyuan Zhou ◽

Dion Lepp ◽

Jason Carere ◽

Hai Yu ◽

Chengbo Yang ◽

...

Keyword(s):

Quorum Sensing ◽

Clostridium Perfringens ◽

Binding Activity ◽

Necrotic Enteritis ◽

Wild Type ◽

Two Component System ◽

Collagen Binding ◽

Component System ◽

Two Component

Clostridium perfringens causes necrotic enteritis (NE) in poultry. A chromosomal locus (VR-10B) was previously identified in NE-causing C. perfringens strains that encodes an adhesive pilus (NE pilus), along with a two-component system (TCS), designated here as PilRS. While the NE pilus is important in pathogenesis, the role of PilRS remains to be determined. The current study investigated the function of PilRS, as well as the Agr-like quorum-sensing (QS) system and VirSR TCS, in the regulation of pilin production. Isogenic pilR , agrB and virR null mutants were generated from parent strain CP1 by insertional inactivation using the ClosTron system, along with the respective complemented strains. Immunoblotting analyses showed no detectable pilus production in the CP1 pilR mutant, while production in its complement (CP1 pilR +) was greater than wild-type levels. In contrast, pilus production in the agrB and virR mutants was comparable or higher than the wild type, but reduced in their respective complemented strains. When examined for collagen-binding activity, the pilR mutant showed significantly lower binding to most collagen types (types I – V) than CP1 ( p ≤ 0.05), whereas this activity was restored in the complemented strain ( p > 0.05). In contrast, binding of agrB and virR mutants to collagen showed no significant differences in collagen-binding activity compared to CP1 ( p > 0.05), whereas the complemented strains exhibited significantly reduced binding ( p ≤ 0.05). These data suggest that the PilRS TCS positively regulates pilus production in C. perfringens , while the Agr-like QS system may serve as a negative regulator of this operon. Importance Clostridium perfringens type G isolates cause necrotic enteritis (NE) in poultry, presenting a major challenge for poultry production in the post-antibiotic era. Multiple factors in C. perfringens , including both virulent and non-virulent, are involved in the development of the disease. We previously discovered a cluster of C. perfringens genes that encode a pilus involved in adherence and NE development and a predicted two-component regulatory system (TCS), designated PilRS. In the present study, we have demonstrated the role of PilRS in regulating pilus production and collagen binding of C. perfringens . In addition, the Agr-like quorum sensing signalling pathway was found to be involved in the regulation. These findings have identified additional targets for developing non-antibiotic strategies to control NE disease.

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Isolation and characterization of Clostridium perfringens strains isolated from ostriches (Struthio camelus) in Vietnam

Veterinary World ◽

10.14202/vetworld.2020.1679-1684 ◽

2020 ◽

Vol 13 (8) ◽

pp. 1679-1684

Author(s):

Tham Thi Nguyen ◽

Hung Vu-Khac ◽

Tan Duc Nguyen

Keyword(s):

Clostridium Perfringens ◽

Pathogenic Bacteria ◽

Necrotic Enteritis ◽

Fecal Samples ◽

Isolation And Characterization ◽

Sds Page ◽

Pcr Method ◽

Encoding Gene

Background and Aim: Clostridium perfringens can cause enteritis in ostriches. The toxin release is believed to play a major role in determining pathogenesis properties of these pathogenic bacteria. This study was conducted to isolate and characterize C. perfringens strains from ostriches in Vietnam for identifying if particular virulence factors of these pathogenic bacteria are associated with enteritis progress in ostriches. Materials and Methods: The prevalence of cpa, cpb, iA, etx, cpe, and cpb2 genes among C. perfringens isolates was determined by a multiplex polymerase chain reaction (PCR) method. The NetB toxin-encoding gene was detected by PCR and then sequenced to observe their variation. The expression of NetB toxin was checked by SDS-PAGE. Results: A total of 116 C. perfringens isolates were obtained from 318 fecal samples and 105 intestinal organs. Of 80 isolates from fecal samples, 33 isolates were from healthy and 47 isolates were from diseased ostriches. The results of multiplex PCR showed that all 116 C. perfringens strains from healthy and enteric disordered ostriches were positive for the alpha toxin-encoding gene (cpa). The cpe and cpb2 genes were found in only one and five diseased ostriches, respectively. The netB gene was detected in 1/33 (3.03%) C. perfringens isolates from healthy ostriches, in 8/47 (17.05%) isolates from feces, and in 7/36 (19.44%) intestinal contents of diseased ostriches. The full-length sequences of 5 out of 15 netB-positive isolates from diseased ostriches showed 100% identity to each other as well as to the netB sequences available in GenBank. All of these five isolates produced NetB toxin in vitro. Conclusion: Type A is the most prevalent among C. perfringens isolates from ostriches in Vietnam. Especially, the study provides data emphasizing the role of NetB toxin in causing necrotic enteritis by C. perfringens in ostriches.

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Role of Clostridium perfringens Necrotic Enteritis B-like Toxin in Disease Pathogenesis

Vaccines ◽

10.3390/vaccines10010061 ◽

2021 ◽

Vol 10 (1) ◽

pp. 61

Author(s):

Kyung-Woo Lee ◽

Hyun S. Lillehoj

Keyword(s):

Clostridium Perfringens ◽

Large Scale ◽

Scientific Evidence ◽

Short Review ◽

Complex Nature ◽

Necrotic Enteritis ◽

Toxin A ◽

Commercial Broiler ◽

Large Scale Field

Necrotic enteritis (NE) is a devastating enteric disease caused by Clostridium perfringens type A/G that impacts the global poultry industry by compromising the performance, health, and welfare of chickens. Coccidiosis is a major contributing factor to NE. Although NE pathogenesis was believed to be facilitated by α-toxin, a chromosome-encoded phospholipase C enzyme, recent studies have indicated that NE B-like (NetB) toxin, a plasmid-encoded pore-forming heptameric protein, is the primary virulence factor. Since the discovery of NetB toxin, the occurrence of NetB+ C. perfringens strains has been increasingly reported in NE-afflicted poultry flocks globally. It is generally accepted that NetB toxin is the primary virulent factor in NE pathogenesis although scientific evidence is emerging that suggests other toxins contribute to NE. Because of the complex nature of the host-pathogen interaction in NE pathogenesis, the interaction of NetB with other potential virulent factors of C. perfringens needs better characterization. This short review will summarize the primary virulence factors involved in NE pathogenesis with an emphasis on NetB toxin, and a new detection method for large-scale field screening of NetB toxin in biological samples from NE-afflicted commercial broiler flocks.

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Role of Clostridium perfringens Toxins in Necrotic Enteritis in Poultry

Toxinology - Microbial Toxins ◽

10.1007/978-94-007-6449-1_15 ◽

2018 ◽

pp. 119-134

Author(s):

M. Flores-Díaz ◽

E. Barquero-Calvo ◽

M. Ramírez ◽

A. Alape-Girón

Keyword(s):

Clostridium Perfringens ◽

Necrotic Enteritis

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

Animals ◽

10.3390/ani11072027 ◽

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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