Abstract
The aim of this study was to determine the clonal correlation of Campylobacter strains isolated from diarrheal children under 5 years of age in Iran using the PFGE method and to determine the antimicrobial susceptibility and virulence gene content of strains. Of 750 patients with bacterial diarrhea, 33 (4%) Campylobacter spp., including 31 C. jejuni (94%) and 2 C. coli (6%), were isolated during 18-month period in Tehran, Iran. Except for one strain, remaining Campylobacter strains were positive for the flaA gene. A complete set of cytolethal distending toxin (CDT) encoding genes (cdtABC) were detected in 52% of the C. jejuni strains, while the 2 C. coli isolates under study only harbored cdtA and cdtB of the CDT cluster. All strains were resistant to at least three antibiotic classes. Resistance to ampicillin among C. coli and C. jejuni strains was 100% and 84%, respectively, and 80% of all strains were susceptible to gentamicin. PFGE genotyping generated 19 pulsotypes with two major clusters, displaying the maximum and minimum similarity of 100% and 26%, respectively. The C. coli strains showed clearly distinct pulsotypes and each fell within separate clusters. A very homogeneous Campylobacter population was detected among Iranian patients with 33 % of strains showing identical banding patterns and no clear correlation was observed between antibiotic resistance profiles and PFGE patterns of the isolates.
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Alikhani MY, Hashemi SH, Aslani MM, Farajnia S (2013) Prevalence and antibiotic resistance patterns of diarrheagenic Escherichia coli isolated from adolescents and adults in Hamedan, Western Iran. Iran J Microbiol 5:42
Alm RA, Guerry P (1993) Distribution and polymorphism of the flagellin genes from isolates of Campylobacter coli and Campylobacter jejuni. J Bacteriol 175:3051–3057
Asakura M, Samosornsuk W, Taguchi M, Kobayashi K, Misawa N, Kusumoto M, Nishimura K, Matsuhisa A, Yamasaki S (2007) Comparative analysis of cytolethal distending toxin (cdt) genes among Campylobacter jejuni, C. coli and C. fetus strains. Microb Pathog 42:174–83
Bagger-Skjøt L, Sandvang D, Frimodt-Møller N, Lester CH, Olsen KEP, Jannok Porsbo L, Monnet DL, Hammerum AM (2007) Association between antimicrobial resistance and virulence genes in Escherichia coli obtained from blood and faeces. Scand J Infect Dis 39:724–727
Bakhshi B, Kalantar M, Rastegar-Lari A, Fallah F (2016) PFGE genotyping and molecular characterization of Campylobacter spp. isolated from chicken meat. Iran J Vet Res 17:177–183
Boesze-Battaglia K, Brown A, Walker L, Besack D, Zekavat A, Wrenn S, Krummenacher C, Shenker BJ (2009) Cytolethal distending toxin-induced cell cycle arrest of lymphocytes is dependent upon recognition and binding to cholesterol. J Biol Chem 284:10650–10658
CLSI (2010) Clinical and laboratory standards institute. Methods for antimicrobial dilution and disk susceptibility testing of infrequently isolated or fastidious bacteria; approved guideline—second edition (M45-A2). Wayne, PA
Di Giannatale E et al (2014) Characterization of antimicrobial resistance patterns and detection of virulence genes in Campylobacter isolates in Italy. Sensors (Basel) 14:3308–3322. https://doi.org/10.3390/s140203308
Dingle KE, Colles FM, Falush D, Maiden MC (2005) Sequence typing and comparison of population biology of Campylobacter coli and Campylobacter jejuni. J Clin Microbiol 43:340–347. https://doi.org/10.1128/JCM.43.1.340-347.2005
Doran G, Morris D, O'Hare C, DeLappe N, Bradshaw B, Corbett-Feeney G, Cormican M (2005) Cost-effective application of pulsed-field gel electrophoresis to typing of Salmonella enterica serovar Typhimurium. Appl Environ Microbiol 71:8236–8240. https://doi.org/10.1128/AEM.71.12.8236-8240.2005
Eberle KN, Kiess AS (2012) Phenotypic and genotypic methods for typing Campylobacter jejuni and Campylobacter coli in poultry. Poult Sci 91:255–264. https://doi.org/10.3382/ps.2011-01414
Economou V, Gousia P (2015) Agriculture and food animals as a source of antimicrobial-resistant bacteria. Infect Drug Resist 8:49
Eftekhari N, Bakhshi B, Pourshafie MR, Zarbakhsh B, Rahbar M, Hajia M, Ghazvini K (2013) Genetic diversity of Shigella spp. and their integron content. Foodborne Pathog Dis 10:237–242
Engberg J, Aarestrup FM, Taylor DE, Gerner-Smidt P, Nachamkin I (2001) Quinolone and macrolide resistance in Campylobacter jejuni and C. coli: resistance mechanisms and trends in human isolates. Emerg Infect Dis 7:24–34. https://doi.org/10.3201/eid0701.700024
Epps SV, Harvey RB, Hume ME, Phillips TD, Anderson RC, Nisbet DJ (2013) Foodborne Campylobacter: infections, metabolism, pathogenesis and reservoirs. Int J Environ Res Public Health 10:6292–6304. https://doi.org/10.3390/ijerph10126292
Fallon R, O'sullivan N, Maher M, Carroll C (2003) Antimicrobial resistance of Campylobacter jejuni and Campylobacter coli isolates from broiler chickens isolated at an Irish poultry processing plant. Lett Appl Microbiol 36:277–281
Feizabadi MM, Dolatabadi S, Zali MR (2007) Isolation and drug-resistant patterns of Campylobacter strains cultured from diarrheic children in Tehran. Jpn J Infect Dis 60:217–219
Gaul SB, Wedel S, Erdman MM, Harris D, Harris IT, Ferris KE, Hoffman L (2007) Use of pulsed-field gel electrophoresis of conserved XbaI fragments for identification of swine Salmonella serotypes. J Clin Microbiol 45:472–476
Ghorbanalizadgan M, Bakhshi B, Kazemnejad Lili A, Najar-Peerayeh S, Nikmanesh B (2014) A molecular survey of Campylobacter jejuni and Campylobacter coli virulence and diversity. Iran Biomed J 18:158–164
Hald T (2010) Analysis of the baseline survey on the prevalence of Campylobacter in broiler batches and of Campylobacter and Salmonella on broiler carcasses in the EU, 2008, Part A: Campylobacter and Salmonella prevalence estimates. European Food Safety Authority, Parma
Hamidian M, Sanaei M, Azimi-Rad M, Tajbakhsh M, Dabiri H, Zali M-R (2011) fla-typing, RAPD analysis, isolation rate and antimicrobial resistance profile of Campylobacter jejuni and Campylobacter coli of human origin collected from hospitals in Tehran, Iran. Ann Microbiol 61:315–321
Kalantar M, Soltan Dallal M-M, Fallah F, Yektaie F (2017) Monitoring the virulence genes in Campylobacter coli strains isolated from chicken meat in Tehran, Iran. Infect Epidemiol Microbiol 3:12–15
Khoshbakht R, Tabatabaei M, Hosseinzadeh S, Aski HS, Seifi S (2015) 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 8: e18573
Konkel ME, Gray SA, Kim BJ, Garvis SG, Yoon J (1999) Identification of the enteropathogens Campylobacter jejuni and Campylobacter coli based on the cadF virulence gene and its product. J Clin Microbiol 37:510–517
Lis L, Connerton IF (2016) The minor flagellin of Campylobacter jejuni (FlaB) confers defensive properties against bacteriophage infection. Front Microbiol 7:1–12
Luangtongkum T, Jeon B, Han J, Plummer P, Logue CM, Zhang Q (2009) Antibiotic resistance in Campylobacter: emergence, transmission and persistence. Future Microbiol 4:189–200
McGowan-Spicer LL, Fedorka-Cray PJ, Frye JG, Meinersmann RJ, Barrett JB, Jackson CR (2008) Antimicrobial resistance and virulence of Enterococcus faecalis isolated from retail food. J Food Prot 71:760–769
Modirrousta S, Shapouri R, Rezasoltani S, Molaabaszadeh H (2016) Prevalence of Campylobacter spp. and their common serotypes in 330 cases of red-meat, chicken-meat and egg-shell in Zanjan City, Iran. Infection. Epidemiol Microbiol 2:8–10
Mohran ZS, Guerry P, Lior H, Murphy JR, El-Gendy AM, Mikhail MM, Oyofo BA (1996) Restriction fragment length polymorphism of flagellin genes of Campylobacter jejuni and/or C. coli isolates from Egypt. J Clin Microbiol 34:1216–1219
Newell D et al (2011) Biosecurity-based interventions and strategies to reduce Campylobacter spp. on poultry farms. Appl Environ Microbiol 77:8605–8614
Nylen G et al (2002) The seasonal distribution of campylobacter infection in nine European countries and New Zealand. Epidemiol Infect 128:383–390
Oza AN, McKenna JP, McDowell SW, Menzies FD, Neill SD (2003) Antimicrobial susceptibility of Campylobacter spp. isolated from broiler chickens in Northern Ireland. J Antimicrob Chemother 52:220–223. https://doi.org/10.1093/jac/dkg333
Parkar SF et al (2013) Prevalence, seasonality and antibiotic susceptibility of thermophilic Campylobacters in ceca and carcasses of poultry birds in the live-bird market. Afr J Microbiol Res 7:2442–2453
Peters TM (2009) Pulsed-field gel electrophoresis for molecular epidemiology of food pathogens. Methods Mol Biol 551:59–70. https://doi.org/10.1007/978-1-60327-999-4_6
Ribot EM, Fitzgerald C, Kubota K, Swaminathan B, Barrett TJ (2001) Rapid pulsed-field gel electrophoresis protocol for subtyping of Campylobacter jejuni. J Clin Microbiol 39:1889–1894. https://doi.org/10.1128/JCM.39.5.1889-1894.2001
Serichantalergs O et al (2010) PFGE, Lior serotype, and antimicrobial resistance patterns among Campylobacter jejuni isolated from travelers and US military personnel with acute diarrhea in Thailand, 1998-2003. Gut Pathog 2:1–11
Shams S, Bakhshi B, Moghadam TT (2016a) In silico analysis of the cadF gene and development of a duplex polymerase chain reaction for species-specific identification of Campylobacter jejuni and Campylobacter coli. Jundishapur J Microbiol 9:1–7
Shams S, Bakhshi B, Nikmanesh B (2016b) Designing a rapid and accurate method for transportation and culture of the Campylobacter jejuni and Campylobacter coli-fastidious bacteria in the children with bacterial gastrointestinal symptoms. Koomesh J 18:71–78
Shams S, Ghorbanalizadgan M, Haj Mahmmodi S, Piccirillo A (2017) Evaluation of a multiplex PCR assay for the identification of Campylobacter jejuni and Campylobacter coli. Infect Epidemiol Med 3:6–8
Siemer B, Nielsen E, On S (2005) Identification and molecular epidemiology of Campylobacter coli isolates from human gastroenteritis, food, and animal sources by amplified fragment length polymorphism analysis and Penner serotyping. Appl Environ Microbiol 71:1953–1958
Singh R, Singh P, Rathore R, Dhama K, Malik S (2008) Studies on effect of seasonal variations on the prevalence of Indian. J Comp Microbiol Immunol Infect Dis 29:45–48
Taremi M, Dallal MMS, Gachkar L, MoezArdalan S, Zolfagharian K, Zali MR (2006) Prevalence and antimicrobial resistance of Campylobacter isolated from retail raw chicken and beef meat, Tehran, Iran. Int J Food Microbiol 108:401–403
Tenover FC, Arbeit RD, Goering RV, Mickelsen PA, Murray BE, Persing DH, Swaminathan B (1995) Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing. J Clin Microbiol 33:2233–2239
Van Vliet A, Ketley J (2001) Pathogenesis of enteric Campylobacter infection. J Appl Microbiol 90:45S–56S
Wang Y, Taylor D (1990) Natural transformation in Campylobacter species. J Bacteriol 172:949–955
Wardak S, Jagielski M (2009) Evaluation of genotypic and phenotypic methods for the differentiation of Campylobacter jejuni and Campylobacter coli clinical isolates from Poland. II. PFGE, ERIC-PCR, PCR-flaA-RFLP and MLST. Med Dosw Mikrobiol 61:63–77
Wei B, Cha SY, Kang M, Roh JH, Seo HS, Yoon RH, Jang HK (2014) Antimicrobial susceptibility profiles and molecular typing of Campylobacter jejuni and Campylobacter coli isolates from ducks in South Korea. Appl Environ Microbiol 80:7604–7610. https://doi.org/10.1128/AEM.02469-14
Whiley H, van den Akker B, Giglio S, Bentham R (2013) The role of environmental reservoirs in human campylobacteriosis. Int J Environ Res Public Health 10:5886–5907
Wieczorek K, Osek J (2013) Antimicrobial resistance mechanisms among Campylobacter. Biomed Res Int 2013:1–12
Wieczorek K, Szewczyk R, Osek J (2012) Prevalence, antimicrobial resistance, and molecular characterization of Campylobacter jejuni and C. coli isolated from retail raw meat in Poland. Vet Med (Praha) 57:293–299
Zhang J, Diao B, Zhang N, Cui Z, Zhang L, Xu J, Kan B (2007) Comparison of different electrophoretic parameters of pulse-field gel electrophoresis for Vibrio cholerae subtyping. J Microbiol Methods 71:15–22
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The authors would like to thank the Research Council of the Tarbiat Modares University for the financial support of the study.
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The study was reviewed and approved by Medical Ethics Committee of the Tarbiat Modares University (Code: IR.MODARES.REC) before it began. The written informed consent was provided by participants in order to join in this study.
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Ghorbanalizadgan, M., Bakhshi, B., Shams, S. et al. Pulsed-field gel electrophoresis fingerprinting of Campylobacter jejuni and Campylobacter coli strains isolated from clinical specimens, Iran. Int Microbiol 22, 391–398 (2019). https://doi.org/10.1007/s10123-019-00062-8
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DOI: https://doi.org/10.1007/s10123-019-00062-8