Characterization of β-lactamase and quinolone resistant Clostridium perfringens recovered from broiler chickens with necrotic enteritis in Bangladesh

Document Type : Full paper (Original article)

Authors

1 Animal Health Research Division, Bangladesh Livestock Research Institute, Savar, Dhaka 1341, Bangladesh

2 Nourish Poultry Diseases Diagnostic Laboratory, Nourish Poultry and Hatchery Ltd., Dhaka 1210, Bangladesh

Abstract

Background: Clostridium perfringens causes necrotic enteritis (NE) and is considered a major economic burden in the broiler industry and a significant foodborne pathogen, worldwide. Aims: Clostridium perfringens isolated from NE affected broiler chickens was aimed to characterize and the presence of β-lactamase and quinolone resistant genes were also investigated in the isolates. Methods: A total of 224 intestinal and caecal specimens were collected from NE affected broiler chickens and cultured to isolate C. perfringens. The toxicogenic characterization of C. perfringens was appraised using polymerase chain reaction (PCR) and antibiotic susceptibility testing (disc diffusion method). The selected C. perfringens isolates were characterized for β-lactamase and quinolone encoding genes by PCR analysis. Results: All isolates were cultured positive for C. perfringens and the toxin-encoding genes of C. perfringens (α-, β-, β2-, ε-, ι-, and enterotoxin) were also identified. About 65.6% of isolates had a multi-drug resistant (MDR) profile but none of these isolates were resistant or susceptible to all screened antibiotics. A subset of isolates, 160 and 98 were analyzed for β-lactamase and quinolone genes, respectively, and recognized blaTEM, blaSHV, and blaOXA in 64 (40%; CI: 32.35-48.03%; P<0.001) isolates, and qnrB and qnrS in 28 (28.57%; CI: 19.90-38.58%; P<0.001) isolates except qnrA. Conclusion: Therefore, the isolates of C. perfringens were toxicogenic and carried β-lactamase, and quinolone resistance genes. Nowadays, the rational use of antibiotics and safe production of broiler chickens are the major concern to save public health.

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References

Abrar, S; Ain, NU; Liaqat, H; Hussain, S; Rasheed, F and Riaz, S (2019). Distribution of blaCTX-M, blaTEM, blaSHV and blaOXA genes in extended-spectrum-β-lactamase-producing clinical isolates: A three-year multi-center study from Lahore, Pakistan. Antimicrob. Resist. Infect. Control. 8: 1-10.
Ahmed, I; Rabbi, MB and Sultana, S (2019). Antibiotic resistance in Bangladesh: A systematic review. Int. J. Infect. Dis., 80: 54-61.
Ali, MZ; Islam, E and Giasuddin, M (2019). Outbreak investigation, molecular detection, and characterization of foot and mouth disease virus in the Southern part of Bangladesh. J. Adv. Vet. Anim. Res., 6: 346-354.
Arnold, S; Gassner, B; Giger, T and Zwahlen, R (2004). Banning antimicrobial growth promoters in feedstuffs does not result in increased therapeutic use of antibiotics in medicated feed in pig farming. Pharmacoepidemiol. Drug Saf., 13: 323-331.
Burki, TK (2018). Superbugs: An arms race against bacteria. Lancet Respir. Med., 6: 668.
Casewell, M; Friis, C; Marco, E; McMullin, P and Phillips, I (2003). The European ban on growth-promoting antibiotics and emerging consequences for human and animal health. Antimicrob. Agents Chemother., 52: 159-161.
Cattoir, V; Poirel, L; Rotimi, V; Soussy, CJ and Nordmann, P (2007a). Multiplex PCR for detection of plasmid-mediated quinolone resistance qnr genes in ESBL-producing enterobacterial isolates. J. Antimicrob. Chemother., 60: 394-397.
Cattoir, V; Weill, FX; Poirel, L; Fabre, L; Soussy, CJ and Nordmann, P (2007b). Prevalence of qnr genes in Salmonella in France. J. Antimicrob. Chemother., 59: 751-754.
Chen, J; Rood, JI and McClane, BA (2011). Epsilon-toxin production by Clostridium perfringens type D strain CN3718 is dependent upon the agr operon but not the VirS/VirR two-component regulatory system. MBio. 2: e00275-00311.
CLSI (Clinical and Laboratory Standards Institute) (2012). Methods for antimicrobial susceptibility testing of anaerobic bacteria; approved standard. 8th Edn., Wayne, PACLSICLSI Document M11-A8 2012.
Collier, CT; Van, DKJD; Deplancke, B; Anderson, DB and Gaskins, HR (2003). Effects of tylosin on bacterial mucolysis, Clostridium perfringens colonization, and intestinal barrier function in a chick model of necrotic enteritis. Antimicrob. Agents Chemother., 47: 3311-3317.
Cooper, KK and Songer, JG (2010). Virulence of Clostridium perfringens in an experimental model of poultry necrotic enteritis. Vet. Microbiol., 142: 323-328.
Cooper, KK; Songer, JG and Uzal, FA (2013). Diagnosing clostridial enteric disease in poultry. J. Vet. Diagn. Invest., 25: 314-327.
Dallenne, C; Da Costa, A; Decré, D; Favier, C and Arlet, G (2010). Development of a set of multiplex PCR assays for the detection of genes encoding important β-lactamases in Enterobacteriaceae. J. Antimicrob. Chemother., 65: 490-495.
Diarra, MS; Silversides, FG; Diarrassouba, F; Pritchard, J; Masson, L; Brousseau, R; Bonnet, C; Delaquis, P; Bach, S; Skura, BJ and Topp, E (2007). Impact of feed supplementation with antimicrobial agents on growth performance of broiler chickens, Clostridium perfringens and enterococcus counts, and antibiotic resistance phenotypes and distribution of antimicrobial resistance determinants in Escherichia coli isolates. Appl. Environ. Microbiol., 73: 6566-6576.
Fish and Animal Feed Act (2010). Available at: http://dls.portal.gov.bd/sites/default/files/files/dls.portal.gov.bd/page/b4ea0518_7cf7_43e5_b0b8_756b831991d4/1.%20Fish%20feed%20and%20Animal%20Feed%20Act%20-2010.pdf. Accessed Oct.16.2019.
Freedman, JC; Theoret, JR; Wisniewski, JA; Uzal, FA; Rood, JI and McClane, BA (2015). Clostridium perfringens type A-E toxin plasmids. Res. Microbiol., 166: 264-279.
Garoff, L; Yadav, K and Hughes, D (2017). Increased expression of Qnr is sufficient to confer clinical resistance to ciprofloxacin in Escherichia coli. J. Antimicrob. Chemother., 73: 348-352.
Gkiourtzidis, K; Frey, J; Bourtzi-Hatzopoulou, E; Iliadis, N and Sarris, K (2001). PCR detection and prevalence of α-, β-, β2-, ε-, ι- and enterotoxin genes in Clostridium perfringens isolated from lambs with Clostridialdysentery. Vet. Microbiol., 82: 39-43.
Gyles, C and Boerlin, P (2014). Horizontally transferred genetic elements and their role in pathogenesis of bacterial disease. Vet. Pathol., 51: 328-340.
IBM Corp. Released (2017). IBM SPSS Statistics for Windows, Version 25.0, Armonk, NY: IBM Corp. Available at: https://www.ibm.com/support/pages/how-cite-ibm-spss-statistics-or-earlier-versions-spss. Accessed Oct.16.2019.
Immerseel, FV; Buck, JD; Pasmans, F; Huyghebaert, G; Haesebrouck, F and Ducatelle, R (2004). Clostridium perfringens in poultry: an emerging threat for animal and public health. Avian Pathol., 33: 537-549.
Kim, JB; Kim, JM; Cho, SH; Oh, HS; Choi, NJ and Oh, DH (2011). Toxin genes profiles and toxin production ability of Bacillus cereus isolated from clinical and food samples. J. Food Sci., 76: 25-29.
Landers, TF; Cohen, B; Wittum, TE and Larson, EL (2012). A review of antibiotic use in food animals: perspective, policy, and potential. Public Health Rep., 127: 4-22.
Lee, YJ (2016). Antimicrobial resistance and molecular characterization of Clostridium perfringens isolated from chicken. J. Prev. Vet. Med., 40: 71-79.
Marshall, BM and Levy, SB (2011). Food animals and antimicrobials: impacts on human health. Clin. Microbiol. Rev., 24: 718-733.
Miah, MS; Asaduzzaman, M; Sufian, MA and Hossain, MM (2011). Isolation of Clostridium perfringens, causal agents of necrotic enteritis in chickens. J. Bangladesh Agril. Univ., 9: 97-102.
Miller, RW; Skinner, J; Sulakvelidze, A; Mathis, GF and Hofacre, CL (2010). Bacteriophage therapy for control of necrotic enteritis of broiler chickens experimentally infected with Clostridium perfringens. Avian Dis., 54: 33-40.
M’Sadeq, SA; Wu, S; Swick, RA and Choct, M (2015).
Towards the control of necrotic enteritis in broiler chickens with in-feed antibiotics phasing-out worldwide. Anim. Nutr., 1: 1-11.
Mwangi, S; Timmons, J; Fitz-Coy, S and Parveen, S (2018). Characterization of Clostridium perfringens recovered from broiler chicken affected by necrotic enteritis. Poult. Sci., 98: 128-135.
Nhung, NT; Chansiripornchai, N and Carrique-Mas, JJ (2017). Antimicrobial resistance in bacterial poultry pathogens: a review. Front. Vet. Sci., 10: 126.
Park, JY; Kim, S; Oh, JY; Kim, HR; Jang, I; Lee, HS and Kwon, YK (2015). Characterization of Clostridium perfringens isolates obtained from 2010 to 2012 from chickens with necrotic enteritis in Korea. Poult. Sci., 94: 1158-1164.
Poirel, L; Cattoir, V and Nordmann, P (2012). Plasmid-mediated quinolone resistance; interactions between human, animal, and environmental ecologies. Front. Microbiol., 3: 24.
Prescott, JF (2016). Brief description of animal pathogenic Clostridia. In: Uzal, FA; Songer, JG and Prescott, JF (Eds.), Clostridial diseases of animals. (1st Edn.), Ames, Iowa, John Wiley and Sons. PP: 13-19.
Scallan, E; Hoekstra, RM; Angulo, FJ; Tauxe, RV; Widdowson, MA; Roy, SL; Jones, JL and Griffin, PM (2011). Foodborne illness acquired in the United States—major pathogens. Emerg. Infect. Dis., 17: 7-15.
Singer, RS; Porter, LJ; Thomson, DU; Gage, M; Beaudoin, A and Wishnie, JK (2019). Potential impacts on animal health and welfare of raising animals without antibiotics. BioRxiv. 600965.
Skinner, JT; Bauer, S; Young, V; Pauling, G and Wilson, J (2010). An economic analysis of the impact of subclinical (mild) necrotic enteritis in broiler chickens. Avian Dis., 54: 1237-1240.
The Daily Star (2019). Prescription from vet must for giving antibiotics to cows: HC. Available at: https://www. thedailystar.net/city/without-prescription-no-antibiotics-to-cows-high-court-1772242. Accessed Oct.16.2019.
Uzal, FA; Freedman, JC; Shrestha, A; Theoret, JR; Garcia, J; Awad, MM; Adams, V; Moore, RJ; Rood, JI and McClane, BA (2014). Towards an understanding of the role of Clostridium perfringens toxins in human and animal disease. Future Microbiol., 9: 361-377.
Wade, B and Keyburn, A (2015). The cost of necrotic enteritis is huge. World Poultry News 2015. 31: 5. Available at: https://www.poultryworld.net/Meat/Articles/ 2015/10/The-true-cost-of-necrotic-enteritis-2699819W/. Accessed Oct.16.2019.
Yang, WY; Lee, YJ; Lu, HY; Branton, SL; Chou, CH and Wang, C (2019). The netB-positive Clostridium perfringens in the experimental induction of necrotic enteritis with or without predisposing factors. Poult. Sci., 98: 5297-5306.
Yoo, HS; Lee, SU; Park, KY and Park, YH (1997). Molecular typing and epidemiological survey of prevalence of Clostridium perfringens types by multiplex PCR. J. Clin. Microbiol., 35: 228-232.

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