ANTIMICROBIAL RESISTANCE, VIRULENCE-ASSOCIATED GENES, AND FLAGELLIN TYPING OF THERMOPHILIC Campylobacter SPECIES ISOLATED FROM DIARRHEIC HUMANS, RAW MILK, AND BROILER NICHES
DOI:
https://doi.org/10.26873/SVR-1436-2021Keywords:
Campylobacter species, antimicrobial resistance, virulence genes, flagellin typingAbstract
Animal-to-human transmission is frequently linked to commensal bacteria in the gastrointestinal tract, as Campylobacter species. We provide a better insight into the existence of cytolethal distending toxin (cdt) and flagellinA (flaA) genes in different antimicrobial resistance patterns (pandrug-resistance (PDR), extensive drug-resistance (XDR) and multidrug-resistance (MDR)] in Campylobacter species recovered from chickens, milk, and human sources in Egypt. Campylobacter species isolation rate was 89.44%, being 79.50% and 20.50% for C. jejuni and C. coli, respectively. Animal samples (chickens and raw milk) showed a higher prevalence of C. coli (21.17%); whereas C. jejuni was highly documented in human samples (83.33%). Testing of antimicrobial susceptibilities revealed that none of the examined campylobacters was pansusceptible, while PDR (1.86%), XDR (65.53%), and MDR (32.61%) campylobacters were reported. Molecular analysis revealed that 50%, and13.16% of the isolated campylobacters were positive for the cdt and flaA genes, respectively. Interestingly, all flaA-positive isolates were cdt-positive C. jejuni. Restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) assay revealed a genetic diversity among flaA harboring isolates presented as three different RFLP patterns of varying sizes ranged from 112 to 647 bp, where flaA-RFLP pattern-I is overrepresented in C. jejuni isolates from various origins (human, milk, and chicken sources). Evidence from this study showed the possibility of treatment failure in campylobacteriosis due to the existence of resistant isolates to all antimicrobial drugs (PDR) and the marked genetic variability of flaA-RFLP pattern that is useful in the epidemiological issues.
References
● 1. Epps SV, Harvey RB, Hume ME, Phillips TD, Anderson RC, et al. Foodborne Campylobacter: infections, metabolism, pathogenesis and reservoirs. Int J Environ Res Public Health 2013; 10: 6292-6304.
● 2. Mohamed MEM, Mohamed RE., Gharieb RM, Amin MA and Ahmed HA. Antimicrobial Resistance, Viru-lence associated genes and biofilm formation of Salmonella Species iso-lated from different sources. Zag Vet J (2021); 49: 94-108
● 3. Kramer JM, Frost JA, Bolton FJ, Wareing DR. Campylobacter contami-nation of raw meat and poultry at re-tail sale: identification of multiple types and comparison with isolates from human infection. J Food Prot 2000; 63: 1654-1659.
● 4. Butzler J.P. Campylobacter, from obscurity to celebrity. Clin Microbiol Infect 2004; 10: 868-876.
● 5. Kurinčič M, Berce I, Zorman T, Smole Možina S. The prevalence of multiple antibiotic resistance in Campylobacter spp. from retail poultry meat. Food Technol Biotechnol 2005;43: 157-163
● 6. Melero B, Juntunen P, Hänninen M.L, Jaime I, Rovira J.Tracing Campylobacter jejuni strains along the poultry meat production chain from farm to retail by pulsed-field gel electrophoresis, and the antimicrobial resistance of isolates. Food Microbiol 2012; 32: 124-128.
● 7. Wieczorek K, Kania I, Osek J. Prevalence and antimicrobial resistance of Campylobacter spp. isolated from poultry carcasses in Poland. J Food Prot 2013; 76:1451-1455.
● 8. Abdollahpour N, Zendehbad B, Alipour A, Khayatzadeh J. Wild-bird feces as a source of Campylobacter jejuni infection in children's playgrounds in Iran. Food Control 2015; 50:378-381.
● 9. El-sayed Y. El-Naenaeey, Marwa I. Abd El-Hamid and Eman K. Khalifa. Foodborne Campylobacter species: Taxonomy, Isolation, Virulence At-tributes and Antimicrobial Re-sistance. Zag Vet J (2020); 48: 414-432
● 10. Brink A, Feldman C, Richards G, Moolman J, Senekal M. Emergence of extensive drug resistance (XDR) among Gram-negative bacilli in South Africa looms nearer. SAMJ: S Afri Med J 2008; 98: 586-592.
● 11. Heywood W, Henderson B, Nair SP. Cytolethal distending toxin: creating a gap in the cell cycle. J Med Microbiol 2005; 54: 207-216.
● 12. Hendrixson DR.A phase‐variable mechanism controlling the Campylobacter jejuni FlgR response regulator influences commensalism. Mol Microbiol 2006; 61: 1646-1659.
● 13. Khoshbakht R, Tabatabaei M, Hosseinzadeh S, Aski HS, Seifi S. Genetic characterization of Campylobacter jejuni and C. coli isolated from broilers using flaA PCR-restriction fragment length polymorphism method in Shiraz, Southern Iran. Jundishapur J Microbiol 2015; 8: e18573.
● 14. Thomrongsuwannakij T, Blackall PJ, Chansiripornchai N. A study on Campylobacter jejuni and Campylobacter coli through commercial broiler production chains in Thailand: antimicrobial resistance, the characterization of DNA gyrase subunit A mutation, and genetic diversity by flagellin A gene restriction fragment length polymorphism. Avian Dis 2017; 61: 186-197.
● 15. Kovács JK, Cox A, Schweitzer B, Maróti G, Kovács T, et al. Virulence Traits of Inpatient Campylobacter jejuni Isolates, and a Transcriptomic Approach to Identify Potential Genes Maintaining Intracellular Survival. Microorganisms 2020; 8: 531.
● 16. Ammar AM, El-Naenaeey E-SY, El-Malt R, El-Gedawy AA, Khalifa E, et al. Prevalence, Antimicrobial Susceptibility, Virulence and Genotyping of Campylobacter jejuni with a Special Reference to the Anti-Virulence Potential of Eugenol and Beta-Resorcylic Acid on Some Multi-Drug Resistant Isolates in Egypt. Animals 2021;11: 3.
● 17. Ghorbanalizadgan M, Bakhshi B, Lili AK, Najar-Peerayeh S, Nikmanesh B. A molecular survey of Campylobacter jejuni and Campylobacter coli virulence and diversity. Iran Biomed J 2014; 18: 158-164.
● 18. Lee J, Jeong J, Lee H, Ha J, Kim S, et al. Antibiotic susceptibility, genetic diversity, and the presence of toxin producing genes in Campylobacter isolates from poultry. Int J Environ Res Public Health 2017; 14: 1400.
● 19. Vandepitte J, Verhaegen J, Engbaek K, Rohner P, Piot P, et al. Basic laboratory procedures in clinical bacteriology. J. Vandepitte ... [et al.], 2nd ed. World Health Organization, (2003), https://apps. who.int/iris/handle/10665/42696.
● 20. Quinn PJ CM, Markey B and Carter CR. Campylobacter spp. In: clinical vet-erinary microbiology, Edition 1. Wolfe Publishing, London, (1994); 268-272.
● 21. Wang G, Clark CG, Taylor TM, Pucknell C, Barton C, et al. Colony multiplex PCR assay for identification and differentiation of Campylobacter jejuni, C. coli, C. lari, C. upsaliensis, and C. fetus subsp. fetus. J Clin Microbiol 2002; 40: 4744-4747.
● 22. Shin E.J , Lee Y.H. Comparison of three different methods for Campylobacter isolation from porcine intestines. J Microbiol Biotechnol 2009; 19: 647-650.
● 23. Bayer A.W, Kirby W.M, Sherris J.C, Turck M. Antibiotic susceptibility testing by a standardized single disc method. Am J Clin Pathol 1966; 45: 493-496.
● 24. CLSI Performance Standards for Antimicrobial Susceptibility Testing. 26th ed. CLSI supplement M100S, (2016): Wayne, PA. Clinical and Laboratory Standards Institute.
● 25. Testing EcoAS. Breakpoint tables for interpretation of MICs and zone diameters. Version 7.1, valid from 2017-03-10. (2017): URL: http://www eucast.org/fileadmin/src/media/PDFs/EUCAST_files/Breakpoint_tables/v_71_Breakpoint_Tables pdf (1211 2017).
● 26. Tambekar D, Dhanorkar D, Gulhane S, Khandelwal V, Dudhane M. Antibacterial susceptibility of some urinary tract pathogens to commonly used antibiotics. Afr J Biotechnol 2006; 5:1684-5315
● 27. Magiorakos AP, Srinivasan A, Carey RB, Carmeli Y, Falagas ME, et al. Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clin Microbiol Infect 2012; 18: 268-281.
● 28. Scarcelli E, Piatti RM, Harakava R, Miyashiro S, Campos F, et al. Use of pcr-rflp of thefla a gene for detection and subtyping of Campylobacter jejuni strains Potentially related to Guillain-barré syndrome, isolated from humans and animals. Braz J Microbiol 2009; 40: 952-959.
● 29. Bang DD, Nielsen EM, Scheutz F, Pedersen K, Handberg K, et al. PCR detection of seven virulence and toxin genes of Campylobacter jejuni and Campylobacter coli isolates from Danish pigs and cattle and cytolethal distending toxin production of the isolates. J Appl Microbiol 2003; 94: 1003-1014.
● 30. Mshana S, Joloba M, Kakooza A, Kaddu-Mulindwa D. Campylobacter spp among Children with acute diarrhea attending Mulago hospital in Kampala-Uganda. Afr Health Sci 2009; 9: 201-205.
● 31. Ammar A M, Abd El-Aziz N K, Elgdawy A A, Emara M S, Hamdy M M. Genotyping and antimicrobial re-sistance of Campylobacter jejuni: A re-view. Adv Anim Vet Sci 2019; 7: 129-136.
● 32. Friedman CR, Hoekstra RM, Samuel M, Marcus R, Bender J, et al. Risk factors for sporadic Campylobacter infection in the United States: a case-control study in FoodNet sites. Clin Infect Dis 2004; 38: S285-S296.
● 33. Neimann J, Engberg J, Mølbak K, Wegener HC. A case–control study of risk factors for sporadic Campylobacter infections in Denmark. Epidemiol Infect 2003; 130: 353-366.
● 34. Abushahba MF, Ahmed S, Ibrahim A, Mosa H. Prevalence of zoonotic species of Campylobacter in broiler chicken and humans in Assiut governorate, Egypt. Approaches in Poultry, Dairy and Veterinary Science 2018; 3: 260-268.
● 35. Jouahri M, Asehraou A, Karib H, Hakkou A, Touhami M. Prevalence and control of thermo-tolerant Campylobacter species in raw poultry meat in Morocco. Meso: prvi hrvatski časopis o mesu 2007; 9: 262-267.
● 36. Jun W, Guo YC, Ning L. Prevalence and risk assessment of Campylobacter jejuni in chicken in China. Biomed Environ Sci 2013; 26: 243-248.
● 37. Abd El-Aziz NK, Ammar AM, Hamdy MM, Gobouri AA, Azab E, et al. First report of aacC5-aadA7Δ4 gene cassette array and phage tail tape measure protein on class 1 integrons of Campylobacter species isolated from animal and human sources in Egypt. Animals 2020;10: 2067.
● 38. Leedom JM. Milk of nonhuman origin and infectious diseases in humans. Clin Infect Dis 2006; 43:610-615.
● 39. Wicker C, Giordano M, Rouger S, Sorin M.L, Arbault P. Campylobacter detection in food using an ELISA based method. Int J Med Microbiol 2001; 291: 1-12.
● 40. Hassanain NA. Antimicrobial resistant Campylobacter jejuni isolated from humans and animals in Egypt. Glob Vet 2011; 6: 195-200.
● 41. El-Hamid MI, El-Aziz A, Khairy N, Samir M, Remela A, et al. Genetic diversity of Campylobacter jejuni isolated from avian and human sources in Egypt. Front Microbiol 2019; 10: 2353.
● 42. De Vries SP, Vurayai M, Holmes M, Gupta S, Bateman M, et al. Phylogenetic analyses and antimicrobial resistance profiles of Campylobacter spp. from diarrhoeal patients and chickens in Botswana. PloS One 2018; 13: e0194481
● 43. Ghunaim H, Behnke JM, Aigha I, Sharma A, Doiphode SH, et al. Analysis of resistance to antimicrobials and presence of virulence/stress response genes in Campylobacter isolates from patients with severe diarrhoea. PLoS One 2015; 10: e0119268.
● 44. Mdegela R.H, Nonga H.E, Ngowi H.A, Kazwala R.R. Prevalence of thermophilic Campylobacter infections in humans, chickens and crows in Morogoro, Tanzania. J Vet Med, Series B2006; 53: 116-121.
● 45. glesias-Torrens Y, Miró E, Guirado P, Llovet T, Muñoz C, et al. Population structure, antimicrobial resistance, and virulence-associated genes in Campylobacter jejuni isolated from three ecological niches: gastroenteritis patients, broilers, and wild birds. Front Microbiol 2018; 9: 1676.
● 46. Shin E, Hong H, Oh Y, Lee Y. First report and molecular characterization of a Campylobacter jejuni isolate with extensive drug resistance from a travel-associated human case. Antimicrob Agents and Chemother 2015; 59: 6670-6672.
● 47. Rozynek E, Dzierzanowska-Fangrat K, Jozwiak P, Popowski J, Korsak D, et al. Prevalence of potential virulence markers in Polish Campylobacter jejuni and Campylobacter coli isolates obtained from hospitalized children and from chicken carcasses. J Med Microbiol 2005; 54: 615-619.
● 48. Fox JG, Rogers AB, Whary MT, Ge Z, Taylor NS, et al. Gastroenteritis in NF-κB-deficient mice is produced with wild-type Camplyobacter jejuni but not with C. jejuni lacking cytolethal distending toxin despite persistent colonization with both strains. Infect Immun 2004;72: 1116-1125.
● 49. Jain D, Prasad KN, Sinha S, Husain N. Differences in virulence attributes between cytolethal distending toxin positive and negative Campylobacter jejuni strains. J Med Microbiol (2008); 57: 267-272.
● 50. Findik A, Ica T, Onuk EE, Percin D, Kevenk TO, et al. Molecular typing and cdt genes prevalence of Campylobacter jejuni isolates from various sources. Trop Anim Health Prod (2011); 43: 711-719.
● 51. Steinhauserova I, Ceskova J, Nebola M. PCR/restriction fragment length polymorphism (RFLP) typing of human and poultry Campylobacter jejuni strains. Letters appl Microbiol (2002); 34: 354-358.
● 52. Tsai H-J, Huang H-C, Tsai H-L, Chang C-C. PCR-based restriction fragment length polymorphism (RFLP) analysis of Campylobacter jejuni isolates from humans, chickens and dogs in northern Taiwan. J Vet Med Sci (2006); 68: 815-819.
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