Nebahat BİLGE, Çiğdem SEZER, Leyla VATANSEVER, Sevda PEHLİVANLAR ÖNEN

Tavuk Etlerinde Sefalosporine Dirençli Escherichia coli Varlığı ve Moleküler Karakterizasyonu

Etlerin geniş spektrumlu beta-laktamaz (GSBL) ve/veya AmpC tipi beta-laktamaz üreten Escherichia coli ile kontaminasyonu, insanlarda meydana gelen enfeksiyonların insidensinde artışa neden olabilir. Bu nedenle bu bakterilerin düzenli olarak taranması, halk sağlığının korunması açısından önemlidir. Bu çalışmada Kars ilinden temin edilen100 adet tavuk eti incelenmiş ve bunlardan 72’sinden 214 adet E. coli izolatı elde edilmiştir. Yapılan antibiyotik duyarlılık testleri izolatlardan 33’ünün (%15.42) en az bir antibiyotiğe dirençli olduğunu, 20’sinin (%9.34) beta-laktamaz, 5’inin ise (%2.33) GSBL ürettiğini göstermiştir. Moleküler testler, beta-laktamaz üreten E. coli izolatlarından 8’inin blaTEM, 3’ünün blaSHV, 7’sinin blaCTX-M ve 13’ünün plazmid aracılı AmpC beta-laktamaz (blaCMY) genlerine sahip olduğuna işaret etmiştir. Plazmid aracılı kinolon direnç genleri de polimeraz zincir reaksiyonu ile görüntülenip sekanslanarak identifiye edilmiş; 3 izolatın qnrB, 4 izolatın da qnrS taşıdığı belirlenmiştir. Bunun yanında izolatlardan 19’unun fimH ve 2’sinin kpsMT II virulens genlerine sahip olduğu görülmüştür. Son olarak 13, 2, 2, 2 ve 1 izolatın sırasıyla filogrup D2, A0, A1, B1 ve D1’e dahil olduğu ortaya konmuştur. Bu çalışmadan elde edilen bulgular, kanatlı etinin GSBL pozitif E. coli açısından önemli bir taşıyıcı rolü üstlenebileceğini göstermiştir.

Occurrence and Molecular Characterization of Cephalosporin Resistant Escherichia coli Isolates from Chicken Meat

Contamination of retail meat with extended spectrum beta-lactamase (ESBL) and/or AmpC type beta-lactamase (AmpC) producing Escherichia coli may contribute to increased incidences of infections in humans. Regular monitoring of these bacteria is required in the view of one health approach. In this study, 100 chicken meat samples obtained from Kars, Turkey were analysed and 214 isolates recovered from 72/100 samples were identified as E. coli. Antibiotic susceptibility tests showed that 15.42% of isolates (33/214) were resistant to at least one cephalosporin antibiotic, 20 (9.34%) were beta-lactamase producer. Among beta-lactamase producing E. coli isolates 8 had blaTEM, 7 had blaCTX-M and 3 had blaSHV genes. Plasmid-mediated AmpC beta-lactamase (blaCMY) gene was present in 13 isolates. Plasmid mediated quinolone resistance genes were also screened by polymerase chain reaction and identified by sequencing of the isolates. As a result, 3 isolates were found to be positive for qnrB, whereas the qnrS gene was detected in 4 isolates. Regarding the virulence genes 19 isolates were positive for fimH and 2 isolates were carrying kpsMT II. Phylo-group D2, A0, A1, B1 and D1 were detected in 13, 2, 2, 2 and 1 isolates, respectively. Our findings indicate that poultry meat could be an important carrier of ESBL positive E. coli.

PDF

___

1. Smet A, Martel A, Persoons D, Dewulf J, Heyndrickx M, Catry B, Herman L, Haesebrouck F, Butaye P: Diversity of extended-spectrum beta-lactamases and class C beta-lactamases among cloacal Escherichia coli isolates in Belgian broiler farms. Antimicrob Agents Chemother, 52 (4): 1238-1243, 2008. DOI: 10.1128/AAC.01285-07
2. Pitout JDD: Extraintestinal pathogenic Escherichia coli: A combination of virulence with antibiotic resistance. Front Microbiol, 3:9, 2012. DOI: 10.3389/ fmicb.2012.00009
3. Clements A, Young JC, Constantinou N, Frankel G: Infection strategies of enteric pathogenic Escherichia coli. Gut Microbes, 3 (2): 71-87, 2012. DOI: 10.4161/gmic.19182
4. EFSA Panel on Biological Hazards (BIOHAZ): Scientific opinion on the public health risks of bacterial strains producing extended-spectrum beta-lactamases and/or AmpC β‐lactamases in food and food producing animals. EFSA J, 9:2322, 2011. DOI: 10.2903/j.efsa.2011.2322
5. Hao H, Cheng G, Iqbal Z, Ai X, Hussain HI, Huang L, Dai M, Wang Y, Liu Z, Yuan Z: Benefits and risks of antimicrobial use in food-producing animals. Front Microbiol, 5:288, 2014. DOI: 10.3389/fmicb.2014.00288
6. Garcia-Graells C, Botteldoom N, Dierick K: Microbial surveillance of ESBL E. coli in poultry meat, a possible vehicle for transfer of antimicrobial resistance to humans. WIV-ISP, 13, 1-6, 2012.
7. Hawkey PM, Jones AM: The changing epidemiology of resistance. J Antimicrob Chemother, 64 (Suppl. 1): i3-i10, 2009. DOI: 10.1093/jac/ dkp256
8. de Kraker MEA, Davey PG, Grundmann H: Mortality and hospital stay associated with resistant Staphylococcus aureus and Escherichia coli bacteraemia: Estimating the burden of antibiotic resistance in Europe. PLoS Med, 8 (10):e1001104, 2011. DOI: 10.1371/journal.pmed.1001104
9. Livermore DM: Has the era of untreatable infections arrived? J Antimicrob Chemother, 64 (Suppl. 1): i29-i36, 2009. DOI: 10.1093/jac/dkp255
10. Smet A, Martel A, Persoons D, Dewulf J, Heyndrickx M, Herman L, Haesebrouck F, Butaye P: Broad-spectrum β-lactamases among Enterobacteriaceae of animal origin: Molecular aspects, motility and impact on public health. FEMS Microbiol Rev, 34 (3): 295-316, 2010. DOI: 10.1111/j.1574-6976.2009.00198.x
11. Aslantaş Ö, Elmacıoğlu S, Yılmaz EŞ: Prevalence and characterization of ESBL- and AmpC-producing Escherichia coli from cattle. Kafkas Univ Vet Fak Derg, 23, 63-67, 2017. DOI: 10.9775/kvfd.2016.15832
12. Projahn M, von Tippelskirch P, Semmler T, Guenther S, Alter T, Roesler U: Contamination of chicken meat with extended spectrum betalactamases producing- Klebsiella pneumoniae and Escherichia coli during scalding and defeathering of broiler carcasses. Food Microbiol, 77, 185-191, 2019. DOI: 10.1016/j.fm.2018.09.010
13. Ewers C, Bethe A, Semmler T, Guenther S, Wieler LH: Extendedspectrum β-lactamase-producing and AmpC-producing Escherichia coli from livestock and companion animals, and their putative impact on public health: A global perspective. Clin Microbiol Infect, 18, 646-655, 2012. DOI: 10.1111/j.1469-0691.2012.03850.x
14. Kluytmans JAJW, Overdevest ITMA, Willemsen I, Kluytmansvan den Bergh MFQ, van der Zwaluw K, Heck M, Rijnsburger M, Vanderbroucke-Grauls CMJE, Savelkoul PHM, Johnston BD, Gordon D, Johnson JR: Extended-spectrum beta-lactamase producing Escherichia coli from retail chicken meat and humans: Comparison of strains, plasmids, resistance genes, and virulence factors. Clin Infect Dis, 56, 478-487, 2013. DOI: 10.1093/cid/cis929
15. Overdevest I, Willemsen I, Rijnsburger M, Eustace A, Xu L, Hawkey PM, Heck M, Savelkoul P, Vandenbroucke-Grauls C, van der Zwaluw K, Huijdens X, Kluytmans J: Extended-spectrum β-lactamase genes of Escherichia coli in chicken meat and humans, the Netherlands. Emerg Infect Dis, 17 (7): 1216-1222, 2011. DOI: 10.3201/eid1707.110209
16. EFSA: Scientific opinion on the public health hazards to be covered by inspection of meat (poultry). EFSA J, 10 (6): 2741, 2012. DOI: 10.2903/j. efsa.2012.2741
17. Ben Said L, Jouni A, Klibi N, Dziri R, Alonso CA, Boudabous A, Ben Slama K, Torres C: Detection of extended spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae in vegetables, soil and water of the farm environment in Tunisia. Int J Food Microbiol, 203, 86-92, 2015. DOI: 10.1016/j.ijfoodmicro.2015.02.023
18. Fattahi F, Mirvaghefi A, Farahmand H, Rafiee G, Abdollahi A: Development of 16S rRNA targeted PCR method for detection of Escherichia coli in Rainbow Trout (Onchorynchus mykiss). Iranian J Pathol, 8 (1): 36-44, 2013.
19. Clinical and Laboratory Standards Institute (CLSI): Performance standards for antimicrobial susceptibility testing. Twenty-fourth informational supplement. M100-S24, 2014.
20. Clermont O, Bonacorsi S, Bingen E: Rapid and simple determination of the Escherichia coli phylogenetic group. Appl Environ Microbiol, 66, 45554558, 2000.
21. Escobar-Páramo P, Le Menac’h A, Le Gall T, Amorin C, Gouriou S, Picard B, Skurnik D, Denamur E: Identification of forces shaping the commensal Escherichia coli genetic structure by comparing animal and human isolates. Environ Microbiol, 8, 1975-1984, 2006. DOI: 10.1111/j.14622920.2006.01077.x
22. Hasman H, Mevius D, Veldman K, Olesen I, Aarestrup FM: β-Lactamases among extended-spectrum β-lactamase (ESBL)-resistant Salmonella from poultry, poultry products and human patients in the Netherlands. J Antimicrob Chemother, 56, 115-121, 2005. DOI: 10.1093/ jac/dki190
23. Leinberger DM, Grimm V, Rubtsova M, Weile J, Schröppel K, Wichelhaus TA, Knabbe C, Schmid RD, Bachmann TT: Integrated detection of extended-spectrum-beta-lactam resistance by DNA microarraybased genotyping of TEM, SHV, and CTX-M genes. J Clin Microbiol, 48, 460471, 2010. DOI: 10.1128/JCM.00765-09
24. Mulvey MR, Soule G, Boyd D, Demczuk W, Ahmed R, Multiprovincial Salmonella Typhimurium Case Control Study Group: Characterization of the first extended-spectrum beta-lactamase-producing Salmonella isolate identified in Canada. J Clin Microbiol, 41, 460-462, 2003. DOI: 10.1128/ JCM.41.1.460-462.2003
25. Cattoir V, Poirel L, Rotimi V, Soussy CJ, Nordmann P: Multiplex PCR for detection of plasmid-mediated quinolone resistance qnr genes in ESBLproducing enterobacterial isolates. J Antimicrob Chemother, 60, 394-397, 2007. DOI: 10.1093/jac/dkm204
26. Cavaco LM, Frimodt-Møller N, Hasman H, Guardabassi L, Nielsen L, Aarestrup FM: Prevalence of quinolone resistance mechanisms and associations to minimum inhibitory concentrations in quinolone-resistant Escherichia coli isolated from humans and swine in Denmark. Microb Drug Resist, 14, 163-169, 2008. DOI: 10.1089/mdr.2008.0821
27. Cavaco LM, Hasman H, Xia S, Aarestrup FM: qnrD, a novel gene conferring transferable quinolone resistance in Salmonella enterica Serovar Kentucky and Bovismorbificans strains of human origin. Antimicrob Agents Chemother, 53, 603-608, 2009. DOI: 10.1128/AAC.00997-08
28. Jacoby GA, Gacharna N, Black TA, Miller GH, Hooper DC: Temporal appearance of plasmid-mediated quinolone resistance genes. Antimicrob Agents Chemother, 53, 1665-1666, 2009. DOI: 10.1128/AAC.01447-08
29. Park CH, Robicsek A, Jacoby GA, Sahm D, Hooper DC: Prevalence in the United States of aac (6’)-Ib-cr encoding a ciprofloxacin-modifying enzyme. Antimicrob Agents Chemother, 50, 3953-3955, 2006. DOI: 10.1128/ AAC.00915-06
30. Chapman TA, Wu XY, Barchia I, Bettelheim KA, Driesen S, Trott D, Wilson M, Chin JJ: Comparison of virulence gene profiles of Escherichia coli strains isolated from healthy and diarrheic swine. Appl Environ Microbiol, 72, 4782-4795, 2006. DOI: 10.1128/AEM.02885-05
31. Kar D, Bandyopadhyay S, Bhattcharyya D, Samanta I, Mahanti A, Nanda PK, Mondal B, Dandapat P, Das AK, Dutta TK, Bandyopadhyay S, Singh RK: Molecular and phylogenetic characterization of multidrug resistant extended spectrum beta-lactamase producing Escherichia coli isolated from poultry and cattle in Odisha, India. Infect Genet Evol, 29, 82-90, 2015. DOI: 10.1016/j.meegid.2014.11.003
32. Kürekci C, Osek J, Aydın M, Tekeli İO, Kurpas M, Wieczorek K, Sakin F: Evaluation of bulk tank raw milk and raw chicken meat samples as source of ESBL producing Escherichia coli in Turkey: Recent insights. J Food Saf, 39:e12605, 2019. DOI: 10.1111/jfs.12605
33. Shigemura H, Matsui M, Sekizuka T, Onozuka D, Noda T, Yamashita A, Kuroda M, Suzuki S, Kimura H, Fujimoto S, Oishi K, Sera N, Inoshima Y, Murakami K: Decrease in the prevalence of extended-spectrum cephalosporin-resistant Salmonella following cessation of ceftiofur use by the Japanese poultry industry. Int J Food Microbiol, 274, 45-51, 2018. DOI: 10.1016/j.ijfoodmicro.2018.03.011
34. Tansawai U, Sanguansermsri D, Na-udom A, Walsh TR, Niumsup PR: Occurrence of extended spectrum β-lactamase and AmpC genes among multidrug-resistant Escherichia coli and emergence of ST131 from poultry meat in Thailand. Food Control, 84, 159-164, 2018. DOI: 10.1016/j. foodcont.2017.07.028
35. Linscott AJ, Brown WJ: Evaluation of four commercially available extended-spectrum beta-lactamase phenotypic confirmation tests. J Clin Microbiol, 43 (3): 1081-1085, 2005. DOI: 10.1128/JCM.43.3.1081-1085.2005
36. Drieux L, Brossier F, Sougakoff W, Jarlier V: Phenotypic detection of extended-spectrum beta-lactamase production in Enterobacteriaceae: Review and bench guide. Clin Microbiol Infect, 14 (Suppl. 1): 90-103, 2008. DOI: 10.1111/j.1469-0691.2007.01846.x
37. Ojo OE, Schwarz S, Michael GB: Detection and characterization of extended-spectrum β-lactamase-producing Escherichia coli from chicken production chains in Nigeria. Vet Microbiol, 194, 62-68, 2016. DOI: 10.1016/j. vetmic.2016.04.022
38. Kim YJ, Moon JS, Oh DH, Chon JW, Song BR, Lim JS, Heo EJ, Park HJ, Wee SH, Sung K: Genotypic characterization of ESBL-producing E. coli from imported meat in South Korea. Food Res Int, 107, 158-164, 2018. DOI: 10.1016/j.foodres.2017.12.023
39. Müller A, Jansen W, Grabowski NT, Monecke S, Ehricht R, Kehrenberg C: ESBL- and AmpC-producing Escherichia coli from legally and illegally imported meat: Characterization of isolates brought into the EU from third countries. Int J Food Microbiol, 283, 52-58, 2018. DOI: 10.1016/j.ijfoodmicro.2018.06.009
40. Tekiner İH, Özpınar H: Occurence and characteristics of extended spectrum beta-lactamase-producing Enterobacteriaceae from foods of animal origin. Braz J Microbiol, 47, 444-451, 2016. DOI: 10.1016/j.bjm.2015.11.034