Farklı Hayvan Dışkılarından Antibiyotik Dirençlilik Özelliklerinin Fenotipik ve Genotipik İncelenmesi
Abstract
Kırıkkale İl’i ve çevresindeki koyun, keçi ve sığır dışkılarından izole edilen Escherichia coli (E. coli) izolatlarının antibiyotik dirençliliği, geniş spektrumlu Beta-laktamaz (GSβL), Ampicilin-C (Amp-C) ve Karbapenemaz aktiviteleri fenotipik ve genotipik olarak incelenmiştir. Toplanan 30 sığır, 30 koyun ve 30 keçi izolatında, Kirby-Bauer disk difüzyon yöntemi ile çoklu ilaç direnci araştırılmıştır. Fenotipik olarak GSβL, Amp-C ve Karbapenemaz aktiviteleri Clinical Laboratory Standards Institute (CLSI) doğrulama testleriyle değerlendirilmiş; GSβL genleri (CTX-M1, OXA-1, CTX-M9, SHV, TEM), Amp-C geni (FOX) ve Karbapenemaz genleri (IMP, KPC, OXA-48, VIM, NDM) klasik PCR ile incelenmiştir.Testler sonucunda direnç sıklığının sırasıyla koyun, sığır ve keçi izolatlarında en yüksek olduğu; en yüksek direnç oranının ise trimethoprim/sulfamethoxazole ve gentamicin antibiyotiklerine karşı geliştiği gözlenmiştir. Fenotipik olarak 22 (%24,4) GSβL pozitif, 1 (%1,1) karbapenemaz şüpheli izolat belirlenmiş; Amp-C pozitiflik gözlenmemiştir. Genotipik incelemede GSβL üretimi 4 sığır (%13,3), 7 koyun (%23,3) ve 11 keçi (%36,6) izolatında tespit edilmiş, SHV tipi genin yaygın olduğu belirlenmiştir. Karbapenemaz şüpheli sığır izolatı PCR ile negatif bulunmuştur. Sonuçlar, Amp-C ve karbapenemaz aktivitesinin saptanmamasının olumlu olduğunu; ancak yüksek GSβL üretiminin antibiyotik dirençliliğinin yayılımı açısından önemli bir sorun teşkil ettiğini göstermektedir.
Keywords
Ethical Statement
This study does not present any ethical concerns.
Supporting Institution
The authors declared that there is no conflict of interest.
References
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- Akçam FZ, Gönen İ, Kaya O et al. (2004): Hastane infeksiyonu etkeni Enterobakterilerde Beta-laktam antibiyotiklere duyarlılık ve ESBL sıklığının araştırılması. Med J SDU. 11, 6-9.
- Aslantaş Ö, Elmacıoğlu S, Yılmaz EŞ (2017): Prevalence and characterization of ESβL- and AmpC-producing Escherichia coli from cattle. Kafkas Univ Vet Fak Derg. 23, 63-67. DOI: 10.9775/kvfd.2016.15832
- Biz E, Kutlu HH, Tekeli A (2016): Comparison of modified hodge test and inhibitor based tests for detection of Carbapenemase producer Enterobacteriaceae isolates. J Ankara Üniv Fac Med. 69, 151-157. DOI: 10.1501/Tipfak_0000000937
- Clinical and Laboratory Standards Institute (2018) The modified hodge test for suspected Carbapenemase production in Enterobacteriaceae. 28th Edition, 2-4. Wayne, PA, USA.
- Dağlar D, Öngüt G (2012): Extended Spectrum Beta-Lactamases (ESβL) and diagnostic methods. JHVS. 1, 1-9 Dallenne C, Costa A, Decre D et al. (2010): Development of a set of multiplex PCR assays for the detection of genes encoding important β-lactamases in Enterobacteriaceae. J AntimicroChemother. 490-495. DOI: 10.1093/jac/dkp498
- Egbule OS, Yusuf İ (2019): Multiple Antibiotics Resistance s in Escherichia coli isolated from Cattle and Poultry Faeces in Abraka- South Nigeria. Pertanika J Trop Agric Sci. 42, 585-594.
- Gülay Z (2002): Interpretation of Antibiotic Susceptibility Tests. Türk Thorax Derg. 3,75-88. Güven G, Kızıl S (2022): Investigation of antimicrobial resistance of ESβL, Amp -C, and carbapenemase producing E. coli strains in retail poultry meat. Turk J Vet Anim Sci. 46, 788-794. DOI: 10.55730/1300-0128.4254
- Mahmood FR, Ahmed IM (2022): Molecular detection of ESβL, Amp-C, ß- Lactamase Escherichia coli isolated from sheep in Mosul city. Iraqi J Vet Sci. 36, 387-392. DOI: 10.33899/ijvs.2021.130380.1810 Mc Hugh, M.L. (2012): Interrater reliability: the kappa statistic. Biochemia Medica 22 (3): 276-282.
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- Ögedey E, Cömert F, Köktürk F et al. (2016): Extended Spectrum Beta-Lactamase (ESβL) producing Escherichia coli and Klebsiella spp. determination of CTX-M enzymes in isolates. Turkish Society Microbiol. 46, 88-96. DOI: 10.5222/TMCD.2016.088
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- Pehlivanoğlu F, Özturk D, Turutoğlu H (2020): Carriage of plasmidic AmpC, Beta-Lactamase producing Escherichia coli in cattle and sheep and characterization of the isolates in terms of antibiogram profiles, phylogeny and virulence. Kafkas Univ Vet Fak Derg. 26, 469-476. DOI: 10.9775/kvfd.2019.23541
- Pehlivanoğlu F (2019): Beta-Lactamase enzyme diversity of Gram-negative bacteria and the situation in bacteria of anamal origin in Turkey. F U Vet J Health Sci. 33, 113-122.
- Plassard V, Gisbert P, Granier S et al. (2021): Surveillance of Extended Spectrum β-Lactamase, Cephalosporinase and Carbapenemase Producing Gram-negative bacteria in raw milk filters and healthy dairy Cattle in Tree Farms in Ile-de-France, France. Front Vet Sci. 8, 633598. DOI: 10.3389/fvets.2021.633598
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- Shabana II, Al- Enazi AT (2020): Investigation of plasmid-mediated resistance in E. coli isolated from healthy and diarrheic sheep and goats. Saudi J Biol Sci. 27, 788-796. DOI: 10.1016/j.sjbs.2020.01.009
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- Tello M, Ocejo M, Oporto B et al. (2020): Prevalence of Cefotaxime resistant Escherichia coli isolates from healthy cattle and sheep in Northern Spain: phenotypic and genome-based characterization of antimicrobial susceptibility. Appl Environ Microbiol. 86, e00742-20. DOI: 10.1128/AEM.00742-20
- The European Committee on Antimicrobial Susceptibility Testing (EUCAST) (2022): Breakpoint tables for interpretation of MICs and zone diameters. Version 13.0. Access address: https://www.eucast.org/clinical_breakpoints, Access date: 04.02.2025.
- Yıldırım D, Pehlivanoğlu F (2018): Extended Spectrum Beta Lactamase producing Escherichia coli in calves. F U Vet J Health Sci. 32, 155-160.
Phenotypic and genotypic investigation of antibiotic resistance, Extended Spectrum Beta-lactamase, Ampicillin-C and Carbapenemase activity of Escherichia coli isolated from cattle, sheep and goat feces
Abstract
In this study, antibiotic resistance, Extended-Spectrum Beta-Lactamase (ESβL), Ampicillin-C (Amp-C), and Carbapenemase activities were investigated in Escherichia coli (E. coli) isolates obtained from sheep, goat, and cattle feces in Kırıkkale Province. The samples were identified using conventional methods and rapid diagnostic kits, and their resistance profiles were evaluated using the Kirby-Bauer disk diffusion method. Phenotypic tests, according to the guidelines of the Clinical and Laboratory Standards Institute (CLSI), were used to determine ESβL, Amp-C, and Carbapenemase activities. Positive samples were further analyzed genotypically using PCR to detect resistance genes. Resistance was most commonly observed in sheep isolates, followed by cattle and goats, particularly against trimethoprim/sulfamethoxazole and gentamicin. Phenotypic tests revealed 22 (24.4%) ESβL-positive isolates and one suspected Carbapenemase producer, while no Amp-C activity was detected. Genotypic analysis identified ESβL genes in 4 cattle (13.3%), 7 sheep (23.3%), and 11 goat (36.6%) isolates, with the SHV gene being the most prevalent. The single isolate phenotypically suspected of carbapenemase activity tested negative by PCR. Although the absence of Amp-C and carbapenemase activity is reassuring, the significant prevalence of ESβL-producing isolates raises concerns about the spread of antibiotic resistance. This highlights the need for monitoring and prudent antibiotic use in livestock management.
Ethical Statement
This study does not present any ethical concerns.
Supporting Institution
The authors declared that there is no conflict of interest.
References
- REFERENCES Abdelmoktader A, Far ATE (2019): Methods of ESβLs detection in clinical microbiology lab. Virol Immunol J. 3, 000222. DOI: 10.23880/vij-16000222
- Akçam FZ, Gönen İ, Kaya O et al. (2004): Hastane infeksiyonu etkeni Enterobakterilerde Beta-laktam antibiyotiklere duyarlılık ve ESBL sıklığının araştırılması. Med J SDU. 11, 6-9.
- Aslantaş Ö, Elmacıoğlu S, Yılmaz EŞ (2017): Prevalence and characterization of ESβL- and AmpC-producing Escherichia coli from cattle. Kafkas Univ Vet Fak Derg. 23, 63-67. DOI: 10.9775/kvfd.2016.15832
- Biz E, Kutlu HH, Tekeli A (2016): Comparison of modified hodge test and inhibitor based tests for detection of Carbapenemase producer Enterobacteriaceae isolates. J Ankara Üniv Fac Med. 69, 151-157. DOI: 10.1501/Tipfak_0000000937
- Clinical and Laboratory Standards Institute (2018) The modified hodge test for suspected Carbapenemase production in Enterobacteriaceae. 28th Edition, 2-4. Wayne, PA, USA.
- Dağlar D, Öngüt G (2012): Extended Spectrum Beta-Lactamases (ESβL) and diagnostic methods. JHVS. 1, 1-9 Dallenne C, Costa A, Decre D et al. (2010): Development of a set of multiplex PCR assays for the detection of genes encoding important β-lactamases in Enterobacteriaceae. J AntimicroChemother. 490-495. DOI: 10.1093/jac/dkp498
- Egbule OS, Yusuf İ (2019): Multiple Antibiotics Resistance s in Escherichia coli isolated from Cattle and Poultry Faeces in Abraka- South Nigeria. Pertanika J Trop Agric Sci. 42, 585-594.
- Gülay Z (2002): Interpretation of Antibiotic Susceptibility Tests. Türk Thorax Derg. 3,75-88. Güven G, Kızıl S (2022): Investigation of antimicrobial resistance of ESβL, Amp -C, and carbapenemase producing E. coli strains in retail poultry meat. Turk J Vet Anim Sci. 46, 788-794. DOI: 10.55730/1300-0128.4254
- Mahmood FR, Ahmed IM (2022): Molecular detection of ESβL, Amp-C, ß- Lactamase Escherichia coli isolated from sheep in Mosul city. Iraqi J Vet Sci. 36, 387-392. DOI: 10.33899/ijvs.2021.130380.1810 Mc Hugh, M.L. (2012): Interrater reliability: the kappa statistic. Biochemia Medica 22 (3): 276-282.
- Oberle K, Capdeville MJ, Berthe T et al. (2012): Evidence for a complex relationship between antibiotics and antibiotic resistant Escherichia coli from medical patients to a receiving environment. Environ Sci Technol. 46, 1859-1868. DOI: 10.1021/es203399h
- Ögedey E, Cömert F, Köktürk F et al. (2016): Extended Spectrum Beta-Lactamase (ESβL) producing Escherichia coli and Klebsiella spp. determination of CTX-M enzymes in isolates. Turkish Society Microbiol. 46, 88-96. DOI: 10.5222/TMCD.2016.088
- Öztürk CE, Kaya AD, Yücel M et al. (2010): Comparison of some phenotypic tests used in Extended Spectrum Beta-Lactamase detection. ANKEM. 24, 111-116.
- Pehlivanoğlu F, Özturk D, Turutoğlu H (2020): Carriage of plasmidic AmpC, Beta-Lactamase producing Escherichia coli in cattle and sheep and characterization of the isolates in terms of antibiogram profiles, phylogeny and virulence. Kafkas Univ Vet Fak Derg. 26, 469-476. DOI: 10.9775/kvfd.2019.23541
- Pehlivanoğlu F (2019): Beta-Lactamase enzyme diversity of Gram-negative bacteria and the situation in bacteria of anamal origin in Turkey. F U Vet J Health Sci. 33, 113-122.
- Plassard V, Gisbert P, Granier S et al. (2021): Surveillance of Extended Spectrum β-Lactamase, Cephalosporinase and Carbapenemase Producing Gram-negative bacteria in raw milk filters and healthy dairy Cattle in Tree Farms in Ile-de-France, France. Front Vet Sci. 8, 633598. DOI: 10.3389/fvets.2021.633598
- Rawat N, Singh F, Hirpurkar SD et al. (2018): Detection and characterization of Extended-Spectrum Beta-Lactamase Genes (bla TEM and bla SHV) among Beta-Lactam resistant fecal coliforms of dairy cattle from Chhattisgarh, India. Turk J Vet Anim Sci. 42, 503-511. DOI: 10.3906/vet-1706-77
- Shabana II, Al- Enazi AT (2020): Investigation of plasmid-mediated resistance in E. coli isolated from healthy and diarrheic sheep and goats. Saudi J Biol Sci. 27, 788-796. DOI: 10.1016/j.sjbs.2020.01.009
- Silva N, Carvalho I, Currie C et al. (2020): Extended‐Spectrum β‐Lactamase and Carbapenemase‐Producing Enterobacteriaceae in food‐producing animals in Europe. 261-273, Antibiotic Drug Resistance First Edition. DOI: 10.1002/9781119282549.ch12
- Tello M, Ocejo M, Oporto B et al. (2020): Prevalence of Cefotaxime resistant Escherichia coli isolates from healthy cattle and sheep in Northern Spain: phenotypic and genome-based characterization of antimicrobial susceptibility. Appl Environ Microbiol. 86, e00742-20. DOI: 10.1128/AEM.00742-20
- The European Committee on Antimicrobial Susceptibility Testing (EUCAST) (2022): Breakpoint tables for interpretation of MICs and zone diameters. Version 13.0. Access address: https://www.eucast.org/clinical_breakpoints, Access date: 04.02.2025.
- Yıldırım D, Pehlivanoğlu F (2018): Extended Spectrum Beta Lactamase producing Escherichia coli in calves. F U Vet J Health Sci. 32, 155-160.
Details
| Primary Language | Turkish |
|---|---|
| Subjects | Microbiology (Other) |
| Journal Section | Original Article |
| Authors | |
| Publication Date | July 24, 2025 |
| Submission Date | January 16, 2025 |
| Acceptance Date | May 5, 2025 |
| Published in Issue | Year 2025 Volume: 36 Issue: 1 |
