Clostridium perfringens Type A from Broiler Chicken with Necrotic Enteritis in Kashmir Valley, India

Nyrah, Qazi; Wani, S.A.; Nazir, Nageena; Rasool, Shafqat; Beigh, Qusina; Kashoo, Zahid Amin; Hussain, I.; Qureshi, Sabia; Ali, R. A.

doi:10.20546/ijcmas.2017.612.283

Cited by 4 publications

(8 citation statements)

References 11 publications

(12 reference statements)

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“…1). These present findings were in close association with the results of Nyrah et al (2017) who recorded 66 isolates from poultry and further confirmed by 16S rRNA species specific PCR. Similar findings were reported by Dar et al (2017) who confirmed all the isolates of C. perfringens by species specific PCR.…”

Section: Genotypic Identification Of C Perfringens By Using Species supporting

confidence: 91%

“…On toxinotyping of the isolates in different types, it was seen that, out of 43 isolates, 27(62.79%) isolates belonged to C. perfringens type A and 12 (27.90%) isolates belonged to C. perfringens type C. Toxin cpa (alpha) has been indicated as a main virulence factor for the development of NE in poultry and this toxin hydrolyses phospholipids and promotes cellular membrane disorganization which leads to destruction of intestinal mucosa. Nyrah et al (2017) detected total 66 isolates of C. perfringens and showed 100 percent positivity towards cpa toxin while 13 isolates (19.69%) showed positivity for cpb2 toxins. Recently, it was speculated that cpb2 positive C. perfringens may cause focal duodenal necrosis in egglaying chickens (França et al, 2016).…”

Section: Toxinotyping Of C Perfringensmentioning

confidence: 96%

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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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Section: Genotypic Identification Of C Perfringens By Using Species supporting

confidence: 91%

Section: Toxinotyping Of C Perfringensmentioning

confidence: 96%

Toxinotyping of Clostridium perfringens in Poultry from Necrotic Enteritis Cases

Shelke¹

2020

JAR

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“…Thus, 41 isolates were established for cpa gene (402 bp) that signifies C. perfringens type A in the broiler samples. These PCR assays were also reported to be utilized by other researchers in India [ 28 , 31 ].…”

Section: Discussionmentioning

confidence: 95%

“…In this evaluation, DNA was extracted from the pure bacterial culture by boiling method as previously described [28][29][30]. In brief, a loopful of bacterial colonies was taken and suspended in 1.5 mL microcentrifuge tubes containing 200 µL distilled water by gentle vortexing and then centrifuged for 10 min at 14,000× g. After that, the pellet was suspended in 200 µL of TE buffer by gentle vortexing.…”

Section: Molecular Detectionmentioning

confidence: 99%

Investigation of Clostridium perfringens in small-scale commercial broiler flocks in Mymensingh district of Bangladesh

et al. 2021

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Background and Aim: Necrotic enteritis (NE) is one of the most prevalent diseases in broiler poultry caused by Clostridium perfringens connected with significant economic losses. A cross-sectional survey was conducted in Mymensingh district of Bangladesh to assess the prevalence of C. perfringens through toxinotyping molecular assay and confirm the risk factors for NE, including antimicrobial-resistant (AMR) status of the isolates. Materials and Methods: We included 40 small-scale commercial broiler farms randomly selected from two subdistricts of Mymensingh district of Bangladesh. As an individual sample, 240 cloacal swabs, and as a pooled sample, 40 drinking water, 40 workers' hand washing, 40 litter swab, and 40 feed samples were collected and evaluated by culture, biochemical, and molecular assays. A pretested semi-structured interview questionnaire was employed to capture flock-level data on risk factors from the farm owners. The flock-level data on risk factors were assessed through univariable and multivariable logistic regression analyses with p<0.05 was considered statistically significant. Results: Overall flock-level prevalence of C. perfringens was estimated to be 10.3% (95% confidence interval [CI] 7.5- 13.6%). Litter swab (pooled) was found to be highly contaminated with C. perfringens (25.0%, 95% CI: 12.7-41.2%) followed by the cloacal swab (10.4%, 95% CI: 6.9-15.0%) and feed sample (5.0%, 95% CI: 0.6-16.9%). History of coccidia infection (Adjusted odds ratio =33.01, 95% CI: 2.14-507.59, p=0.01) was significantly associated with flock-level C. perfringens infection status. In this study, 78.1% isolates were found as multidrug-resistant as they demonstrated resistance to 3-5 antimicrobial agents. Conclusion: Evidence-based control options need to be taken through the uses of prebiotics and probiotics, biosecurity, and hygienic measurement, including control of coccidia infection, is needed to lessen the NE infection and AMR related to this pathogen in small-scale commercial broiler poultry.

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“…The boiling method was applied for the extraction of DNA from the pure bacterial culture as previously described (Nyrah et al, 2017;Osman et al, 2012). A single loopful of bacterial colonies were taken from the culture media and diluted in 1.5 mL microcentrifuge tubes with 200 µL distilled water through gentle vortexing, and then centrifuged at 14,000×g for 10 min.…”

Section: Molecular Confirmationmentioning

confidence: 99%

Occurrence of Clostridium perfringens in layer flocks of selected districts in Bangladesh: molecular typing, antimicrobial susceptibility

Arif

Sultana

Islam

et al. 2022

Asian Australas. J. Biosci. Biotechnol.

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Necrotic enteritis (NE) is one of the important enteric diseases in poultry caused by Clostridium perfringens which leads to considerable economic losses for poultry raisers. The present research was carried out to estimate prevalence, risk factors and to confirm molecular pattern, antimicrobial susceptibility status of Clostridium perfringens in layer flocks in Mymensingh and Gazipur districts of Bangladesh, where a total of 287 samples were collected that comprised of water (30), workers’ hand washing (30), and feed (30) as environmental pooled samples; cloacal swab (150) and post mortem sample (intestine) (47) as individual samples, and evaluated by basic (culture, biochemical test), and finally toxinotyping multiplex PCR assays. The flock-level data on potential risk factors were collected through semi-structured interviews with the farm owners and evaluated via univariable and multivariable logistic regression analyses where a p-value of <0.05 was considered as statistically significant. Overall flock-level prevalence of Clostridium perfringens was confirmed to be 6.3% (95% confidence interval [CI]: 3.8-9.7%). The occurrence of C. perfringens was found to be higher in the intestine samples (12.8%, 95% CI: 4.8- 25.7%) followed by feed (6.7%, 95% CI: 0.8-22.1%), cloacal swabs (6.0%, 95% CI: 2.8-11.1%) and workers’ hand washing (3.3%, 95% CI: 0.1- 12.2%). History of immune-suppressive disease (like New castle disease/egg drop syndrome/infectious bursal disease) [Odds Ratio (OR) = 22.16, 95% CI: 1.40-349.36, P = 0.027] and flock size more than 3000 birds (OR = 11.16, 95% CI: 1.26-98.25, P = 0.029) were found to be associated with C. perfringens infection status in the layer flocks. In this study, C. perfringens toxinotype A is circulating in layer flocks with 72.2% of isolates were established as multidrug-resistant as they demonstrated resistance between 3 to 6 antimicrobial agents. Evidence-based control measures to be taken namely use of probiotics, prebiotic, organic acids, and plant extracts as natural alternatives including control of immune-suppressive diseases are needed to minimize the NE infection and antimicrobial resistance in low biosecurity layer flocks. Asian Australas. J. Biosci. Biotechnol. 2022, 7(2), 36-49

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Clostridium perfringens Type A from Broiler Chicken with Necrotic Enteritis in Kashmir Valley, India

Cited by 4 publications

References 11 publications

Toxinotyping of Clostridium perfringens in Poultry from Necrotic Enteritis Cases

Toxinotyping of Clostridium perfringens in Poultry from Necrotic Enteritis Cases

Investigation of Clostridium perfringens in small-scale commercial broiler flocks in Mymensingh district of Bangladesh

Occurrence of Clostridium perfringens in layer flocks of selected districts in Bangladesh: molecular typing, antimicrobial susceptibility

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