Molecular detection and characterization of Clostridium perfringens toxin genes causing necrotic enteritis in broiler chickens

Kumar, Nitu; Kumar, Naveen; Karthik, A.

doi:10.1007/s11250-019-01847-9

Cited by 17 publications

(4 citation statements)

References 28 publications

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“…In this study, the cpb 2 gene was identified from C. perfringens strains isolated from chickens, pigeons, camels, and human sources. Our results conform with previously published studies that identified the cpb 2 gene in isolates from chickens [ 56 , 58 ], humans [ 47 ], and camels [ 58 ]. A recent study in Egypt documented a high frequency of the cpb 2 gene from C. perfringens isolates in various sources, including chickens and humans [ 59 ].…”

Section: Discussionsupporting

confidence: 93%

“…The beta2 toxin was found to possess a weaker cytotoxic activity than the beta toxin, despite having similar biological activity [ 54 ]. However, a possible association was reported between enteric disease and the presence of C. perfringens isolates carrying the cpb 2 gene [ 56 , 57 ]. In this study, the cpb 2 gene was identified from C. perfringens strains isolated from chickens, pigeons, camels, and human sources.…”

Section: Discussionmentioning

confidence: 99%

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Genetic Relatedness, Antibiotic Resistance, and Effect of Silver Nanoparticle on Biofilm Formation by Clostridium perfringens Isolated from Chickens, Pigeons, Camels, and Human Consumers

Ahmed

Bayomi

Hamed³

et al. 2022

Veterinary Sciences

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In this study, we determined the prevalence and toxin types of antibiotic-resistant Clostridium perfringens in chicken, pigeons, camels, and humans. We investigated the inhibitory effects of AgNPs on biofilm formation ability of the isolates and the genetic relatedness of the isolates from various sources determined using RAPD-PCR. Fifty isolates were identified using PCR, and all the isolates were of type A. The cpe and cpb2 genes were detected in 12% and 56% of the isolates, respectively. The effect of AgNPs on biofilm production of six representative isolates indicated that at the highest concentration of AgNPs (100 µg/mL), the inhibition percentages were 80.8–82.8%. The RAPD-PCR patterns of the 50 C. perfringens isolates from various sources revealed 33 profiles and four clusters, and the discriminatory power of RAPD-PCR was high. Multidrug-resistant C. perfringens isolates are predominant in the study area. The inhibition of biofilm formation by C. perfringens isolates was dose-dependent, and RAPD-PCR is a promising method for studying the genetic relatedness between the isolates from various sources. This is the first report of AgNPs’ anti-biofilm activity against C. perfringens from chickens, pigeons, camels, and humans, to the best of our knowledge.

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Section: Discussionsupporting

confidence: 93%

Section: Discussionmentioning

confidence: 99%

Genetic Relatedness, Antibiotic Resistance, and Effect of Silver Nanoparticle on Biofilm Formation by Clostridium perfringens Isolated from Chickens, Pigeons, Camels, and Human Consumers

Ahmed

Bayomi

Hamed³

et al. 2022

Veterinary Sciences

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“…In the present study, the overall prevalence of C. perfringens in chicken is 34.5%, but in some countries, particularly Jordan and Egypt, it is higher than 40% [ 20 , 21 ]. Besides, the prevalence of C. perfringens in broiler chicken was 33.3% in our study, which is slightly higher than the recent findings of Praveen Kumar et al [ 22 ] and Zhang et al [ 23 ]. They described the prevalence as 21.97% and 23.1% in broiler chicken in India and China, respectively.…”

Section: Discussioncontrasting

confidence: 84%

Phenotypical Identification and Toxinotyping of Clostridium perfringens Isolates from Healthy and Enteric Disease‐Affected Chickens

Rana

Nizami

Islam

et al. 2023

Veterinary Medicine International

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Clostridium perfringens is a ubiquitous spore-forming anaerobic pathogen that is frequently associated with enteric disease in chickens. Moreover, enterotoxin-producing C. perfringens has high zoonotic potential as well as serious public health concerns due to the emanation of food-borne intoxication. The present study was designed to isolate, identify, and toxinotype C. perfringens from both healthy and cases of necrotic or ulcerative enteritis chickens. A total of 110 samples were collected from July 2019 to February 2021. Among the samples, 38 (34.5%, 95% CI: 26.39–43.83) were positive for C. perfringens and were obtained from broiler 21 (33.3%, 95% CI: 22.91–45.67), Sonali 9 (34.6%, 95% CI: 19.31–53.88), and layer 8 (38%, 95% CI: 20.68–59.20). C. perfringens was highly prevalent (35.7%, 95% CI: 25.48–47.44) in enteritis chickens compared with healthy ones. In multiplex PCR toxinotyping, 34 (89.4%) isolates were identified as C. perfringens type A by the presence of the alpha toxin gene (cpa). Moreover, in addition to the cpa gene, 3 (14.3%, 95% CI: 4.14–35.48) broiler and 1 (11.1%, 95% CI: 0.01–45.67) Sonali isolates harbored the enterotoxin gene (cpe) and were classified as type F. However, none of the isolates carried genes encoding beta (cpb), epsilon (etx), iota (iap), or beta-2 (cpb2) toxins. Multivariable logistic regression analysis identified the following variables such as; “previously used litter materials” (OR 21.77, 95% CI 2.22–212.66, p ≤ 0.008 ); intestinal lesions, “presence of ulceration” (OR 30.01, 95% CI 3.02–297.91, p ≤ 0.004 ); “ballooned with gas” (OR 24.74, 95% CI 4.34–140.86, p ≤ 0.001 ) and “use of probiotics” (OR 5.24, 95% CI 0.74–36.75, p ≤ 0.095 ) act as risk factors for C. perfringens colonization in chicken gut. This is the first study of molecular toxinotyping of C. perfringens from healthy and enteric-diseased chickens in Bangladesh, which might have a potential food-borne zoonotic impact on human health.

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“…A total of 173 samples comprised of intestinal scrapings from dead birds having clinical signs of NE and cloacal swabs from NE suspected live birds were collected from chicken farms located in Anhui ( n = 50), Guangzhou ( n = 56), Guangxi ( n = 48), and Fujian ( n = 19) province, China. The intestinal scrapings from dead chickens were taken using sterile cotton swabs and kept in sealed plastic bags [ 24 ]. The samples were randomly collected from farms having outbreaks of NE located in South China and transported over ice to the lab for processing and stored under refrigerated conditions.…”

Section: Methodsmentioning

confidence: 99%

Animal Model Studies, Antibiotic Resistance and Toxin Gene Profile of NE Reproducing Clostridium perfringens Type A and Type G Strains Isolated from Commercial Poultry Farms in China

et al. 2023

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Poultry necrotic enteritis (NE) is a complex and multifactorial disease caused by Clostridium perfringens types. Earlier, the disease was prevented and/or controlled through the addition of in-feed antibiotics and antimicrobial growth promoters (AGPs). The ban on the use of these agents as feed additives has been a major reason for re-emergence of this disease leading to huge economic losses to the world poultry industry. Understanding the pathogenesis of NE by developing an effective experimental model remains challenging and lacks consistency owing to the involvement of several critical factors involved in causing lesions of disease in the field. In this study, locally characterized C. perfringens types, i.e., ACP (toxinotype A), and GCP (toxinotype G), obtained from NE outbreaks on commercial farms in China (2020–2022), were used to experimentally induce NE in Specific-Pathogen-Free (SPF) chicks. The lesion scores observed on day 20 were 1.9 ± 1.10 (GCP strain) and 1.5 ± 1.08 (ACP strain), and both had significant difference as compared to the control group. The inclusion of fishmeal in addition to oral clostridial dose, i.e., fishmeal (day 7 onward) + Clostridia (7.5 × 108 cfu/mL consecutively for 04 days) induced a lesion score of 2.0 ± 1.15 in respective groups. Use of coccidia (Eimeria necatrix) on day 9 followed by clostridia challenge enhanced the lesion scores to 2.5 ± 1.08 and 2.2 ± 1.23 for type G and type A strains, respectively. When both predisposing factors (coccidia + fish meal) were given together, i.e., fishmeal (day 7 onward) and coccidia (day 9) along with clostridia, the lesion scores were 3.2 ± 1.22 (GCP + coccidia + fish meal) and 3.0 ± 1.15 (ACP + coccidia + fish meal). These results were significantly different from group 1 (ACP) and 2 (GCP), in which only C. perfringens was used to induce NE. The clinical signs as well as histopathological lesions in experimentally induced groups were found similar as reported in the literature. The two type G strains identified in this study were also used for susceptibility testing against various drugs. Both strains were found to be resistant to amikacin, doxycycline, metronidazole, neomycin, nystatin, polymyxin B, streptomycin, and tetracycline. Variable susceptibility was seen against ceftriaxone, florfenicol, gentamicin, and kanamycin drugs. Amoxicillin, ampicillin, cefotaxime, ciprofloxacin, enrofloxacin, ofloxacin, and penicillin were effective drugs based upon their low level of resistance and therefore they might be preferred over other antimicrobial agents for proper treatment/prophylaxis of NE infections. Further studies are needed to study the pathogenesis of NE in detail in experimentally induced models along with continuous monitoring of the resistance pattern of C. perfringens strains in the field.

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Molecular detection and characterization of Clostridium perfringens toxin genes causing necrotic enteritis in broiler chickens

Cited by 17 publications

References 28 publications

Genetic Relatedness, Antibiotic Resistance, and Effect of Silver Nanoparticle on Biofilm Formation by Clostridium perfringens Isolated from Chickens, Pigeons, Camels, and Human Consumers

Genetic Relatedness, Antibiotic Resistance, and Effect of Silver Nanoparticle on Biofilm Formation by Clostridium perfringens Isolated from Chickens, Pigeons, Camels, and Human Consumers

Phenotypical Identification and Toxinotyping of Clostridium perfringens Isolates from Healthy and Enteric Disease‐Affected Chickens

Animal Model Studies, Antibiotic Resistance and Toxin Gene Profile of NE Reproducing Clostridium perfringens Type A and Type G Strains Isolated from Commercial Poultry Farms in China

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