Investigation of Antimicrobial Susceptibilities and Resistance Genes of Campylobacter Isolates from Patients in Edirne, Turkey
,
Abstract
Background: We aimed to determine the susceptibility of Campylobacter isolates obtained from patients to various antimicrobial agents and to investigate some related antimicrobial resistance genes.
Methods: Fifty-six Campylobacter isolates obtained from fecal specimens by conventional methods at the Trakya University Health Center for Medical Research and Practice, Department of Medical Microbiology in Edirne, Turkey, from 2017-2017 were included. Antimicrobial susceptibilities were investigated by the gradient strip test method, and species determination was made by multiplex polymerase chain reaction (mPCR). The presence of the erm(B) gene and tet(O) gene was investigated in all isolates by PCR. DNA sequence analysis was performed to detect the presence of mutations in the 23S rRNA positions 2074 and 2075 in five isolates, including two erythromycin resistant isolates. The gyrA gene mutation was investigated by the mismatch amplification mutation assay (MAMA)-PCR.
Results: In 54 C. jejuni isolates, resistance to erythromycin was 3.7%; to tetracycline, 59.3%; and to ciprofloxacin, 74.1%. Phenotypically, the tet(O) gene was detected in 33 tetracycline-resistant isolates, but no erm(B) gene was found in any of the Campylobacter isolates. As a result of the DNA sequencing, it was found no mutations in the 23S rRNA gene at the 2074 and 2075 positions. The gyrA mutation was observed in all 41 ciprofloxacin resistant Campylobacter isolates.
Conclusion: Among the antimicrobial agents tested, ciprofloxacin had the highest resistance rate, and erythromycin had the lowest. Antimicrobial resistance in Campylobacter increased significantly compared with previously studies in our region as well as in the entire world. Monitoring the resistance to antimicrobial agents used to treat Campylobacter infections is important in determining empiric antimicrobial treatment.
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6. Connell SR, Tracz DM, Nierhaus KH, Tay-lor, DE (2003). Ribosomal protection proteins and their mechanism of tetracy-cline resistance. Antimicrob Agents Chemoth-er, 47 (12): 3675-81.
7. Iovine NM (2013). Resistance mechanisms in Campylobacter jejuni. Virulence, 4 (3): 230-40.
8. Tang Y, Fang L, Xu C, Zhang Q (2017). Antibiotic resistance trends and mecha-nisms in the foodborne pathogen, Cam-pylobacter. Anim Health Res Rev, 18 (2): 87-98.
9. Qin S, Wang Y, Zhang Q, Zhang M, Deng F, Shen Z, Wu C, Wang S, Zhang J, Shen J (2014). Report of ribosomal RNA methylase gene erm(B) in multidrug-resistant Campylobacter coli. J Antimicrob Chemother, 69: 964-8.
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11. Ge B, Wang F, Sjölund-Karlsson M, McDermott PF (2013). Antimicrobial re-sistance in Campylobacter: Susceptibility testing methods and resistance trends. J Microbiol Methods, 95 (1): 57-67.
12. Baker CN (1992). The E‑Test and Campylo-bacter jejuni. Diagn Microbiol Infect Dis, 15: 469‑72.
13. CLSI (2016). Methods for antimicrobial dilution and disk susceptibility testing of infrequently isolat-ed or fastidious bacteria. 3rd ed. CLSI Guide-line M45. Clinical and Laboratory Stand-ards Institute, Wayne, PA.
14. Wang G, Clark CG, Taylor TM, Pucknell C, Barton C, Price L, Woodward DL, Rodgers FG (2002). Colony multiplex PCR assay for identification and differen-tiation of Campylobacter jejuni, C. coli, C. lari, C. upsaliensis, and C. fetus subsp. fetus. J Clin Microbiol, 40 (12): 4744-7.
15. Zhang A, Song L, Liang H, Gu Y, Zhang C, Liu X, Zhang J, Zhang M (2016). Molec-ular subtyping and erythromycin re-sistance of Campylobacter in China. J Appl Microbiol, 121 (1): 287-93.
16. Gibreel A, Tracz DM, Nonaka L, Ngo TM, Connell SR, Taylor DE (2004). Incidence of antibiotic resistance in Campylobacter je-juni isolated in Alberta, Canada, from 1999 to 2002, with special reference to tet(O)-mediated tetracycline resistance. Antimicrob Agents Chemother, 48(9): 3442-50.
17. Zirnstein G, Li Y, Swaminathan B, Angulo F (1999). Ciprofloxacin resistance in Cam-pylobacter jejuni isolates: detection of gyrA resistance mutations by mismatch ampli-fication mutation assay PCR and DNA sequence analysis. J Clin Microbiol, 37 (10): 3276-80.
18. Zirnstein G, Helsel L, Li Y, Swaminathan B, Besser J (2000). Characterization of gyrA mutations associated with fluoroquino-lone resistance in Campylobacter coli by DNA sequence analysis and MAMA PCR. FEMS Microbiol Lett, 190 (1): 1-7.
19. Alonso R, Mateo E, Churruca E, Martinez I, Girbau C, Fernández-Astorga A (2005). MAMA-PCR assay for the detection of point mutations associated with high-level erythromycin resistance in Campylo-bacter jejuni and Campylobacter coli strains. J Microbiol Methods, 63 (1): 99-103.
20. Alfredson DA, Korolik V (2007). Antibiotic resistance and resistance mechanisms in Campylobacter jejuni and Campylobacter coli. FEMS Microbiol Lett, 277 (2): 123-32.
21. Jonaidi-Jafari N, Khamesipour F, Ranjbar R, Kheiri R (2016). Prevalence and antimi-crobial resistance of Campylobacter species isolated from the avian eggs. Food Con-trol, 70: 35-40.
22. Fitzgerald C (2015). Campylobacter. Clin Lab Med, 35(2): 289-98.
23. Patrick ME, Henao OL, Robinson T, Geis-sler AL, Cronquist A, Hanna S, Hurd S, Medalla F, Pruckler J, Mahon BE (2018). Features of illnesses caused by five spe-cies of Campylobacter, Foodborne Dis-eases Active Surveillance Network (FoodNet) - 2010-2015. Epidemiol Infect, 146(1): 1-10.
24. Ciftci N, Turk-Dagi H, Tuncer I (2019). Prevalence and antimicrobial susceptibility of Campylobacter and Salmonella species in patients with acute gastroenteritis. Klimik Derg, 32 (2): 127-31.
25. Ranjbar R, Babazadeh D, Jonaidi-Jafari N (2017). Prevalence of Campylobacter jejuni in adult patients with inflammatory bacterial diarrhea, East Azerbaijan, Iran. Acta Med Mediterr, 33: 901.
26. Zhang L, Li Y, Shao Y, et al (2020). Molecu-lar characterization and antibiotic resistant profiles of Campylobacter species isolated from poultry and diarrheal patients in Southeastern China 2017–2019. Front Mi-crobiol, 11:1244.
27. Sproston EL, Wimalarathna HML, Shep-pard SK (2018). Trends in fluoroquino-lone resistance in Campylobacter. Microb Ge-nom, 4 (8): e000198.
28. Smith JL, Fratamico PM (2010). Fluoroquin-olone resistance in Campylobacter. J Food Prot, 73 (6): 1141-52.
29. Elhadidy M, Ali MM, El-Shibiny A, Miller WG, Elkhatib WF, Botteldoorn N, Dier-ick K. (2020). Antimicrobial resistance patterns and molecular resistance markers of Campylobacter jejuni isolates from human diarrheal cases. PLoS One, 15 (1): e0227833.
30. Kim JS, Lee MY, Kim SJ, Jeon S-E, Cha I, Hong S, Chung GT, Huh M-J, Kang Y-H, Yoo C-K, Kim J (2016). High-level ciprofloxacin-resistant Campylobacter jejuni isolates circulating in humans and animals in Incheon, Republic of Korea. Zoonoses Public Health, 63 (7): 545-54.
31. Kayman T, Abay S, Hızlısoy H (2013). Iden-tification of Campylobacter spp. isolates with phenotypic methods and multiplex polymerase chain reaction and their anti-biotic susceptibilities. Mikrobiyol Bul, 47 (2): 230-9.
32. Kayman T, Abay S, Aydin F, Şahin O (2019). Antibiotic resistance of Campylo-bacter jejuni isolates recovered from hu-mans with diarrhoea in Turkey. J Med Mi-crobiol, 68 (2): 136-42.
33. Zhou J, Zhang M, Yang W, Fang Y, Wang G, Hou F (2016). A seventeen-year ob-servation of the antimicrobial susceptibil-ity of clinical Campylobacter jejuni and the molecular mechanisms of erythromycin-resistant isolates in Beijing, China. Int J Infect Dis, 42: 28-33.
34. Carev M, Kovačić A, Novak A, Tonkić M, Jerončić A (2017). Campylobacter jejuni strains coresistant to tetracycline and ciprofloxacin in patients with gastroenter-itis in Croatia. Infect Dis (Lond), 49 (4): 268-76.
35. Chen J, Sun XT, Zeng Z, Yu YY (2011). Campylobacter enteritis in adult patients with acute diarrhea from 2005 to 2009 in Beijing, China. Chin Med J (Engl), 124 (10): 1508-12.
36. Shen Z, Wang Y, Zhang Q, Shen J (2018). Antimicrobial resistance in Campylobacter spp. Microbiol Spectr, 6 (2).
37. García‑Fernández A, Dionisi AM, Arena S, Iglesias‑Torrens Y, Carattoli A, Luzzi I (2018). Human campylobacteriosis in Ita-ly: Emergence of multi‑drug resistance to ciprofloxacin, tetracycline, and erythro-mycin. Front Microbiol, 9: 1906.
38. Yılmaz AT, Tuğrul HM (2005). Investigation on Campylobacter species as diarrheogenic agents in Edirne, Turkey, and their anti-microbial susceptibility patterns. Turkish J Infect, 19: 53‑9.
39. Eryildiz C, Tabakcioglu K, Kuyucuklu G, Sakru N (2020). Investigation of antimi-crobial resistance and virulence genes of Campylobacter isolates from patients in a tertiary hospital in Edirne, Turkey. Indian J Med Microbiol, 38 (2): 157-61.
2. Wieczorek K, Osek J (2013). Antimicrobial resistance mechanisms among Campylo-bacter. Biomed Res Int, 2013: 340605.
3. Man SM (2011). The clinical importance of emerging Campylobacter species. Nat Rev Gastroenterol Hepatol, 8 (12): 669-85.
4. Whitehouse CA, Zhao S, Tate H (2018). An-timicrobial resistance in Campylobacter spe-cies: Mechanisms and genomic epidemi-ology. Adv Appl Microbiol, 103: 1-47.
5. Igwaran, A, Okoh AI (2019). Human cam-pylobacteriosis: A public health concern of global importance. Heliyon, 5 (11): e02814.
6. Connell SR, Tracz DM, Nierhaus KH, Tay-lor, DE (2003). Ribosomal protection proteins and their mechanism of tetracy-cline resistance. Antimicrob Agents Chemoth-er, 47 (12): 3675-81.
7. Iovine NM (2013). Resistance mechanisms in Campylobacter jejuni. Virulence, 4 (3): 230-40.
8. Tang Y, Fang L, Xu C, Zhang Q (2017). Antibiotic resistance trends and mecha-nisms in the foodborne pathogen, Cam-pylobacter. Anim Health Res Rev, 18 (2): 87-98.
9. Qin S, Wang Y, Zhang Q, Zhang M, Deng F, Shen Z, Wu C, Wang S, Zhang J, Shen J (2014). Report of ribosomal RNA methylase gene erm(B) in multidrug-resistant Campylobacter coli. J Antimicrob Chemother, 69: 964-8.
10. Wang Y, Zhang M, Deng F, Shen Z, Wu C, Zhang J, Zhang Q, Shena J (2014). Emergence of multidrug-resistant Campyl-obacter species isolates with a horizontally acquired rRNA methylase. Antimicrob Agents Chemother, 58 (9): 5405-12.
11. Ge B, Wang F, Sjölund-Karlsson M, McDermott PF (2013). Antimicrobial re-sistance in Campylobacter: Susceptibility testing methods and resistance trends. J Microbiol Methods, 95 (1): 57-67.
12. Baker CN (1992). The E‑Test and Campylo-bacter jejuni. Diagn Microbiol Infect Dis, 15: 469‑72.
13. CLSI (2016). Methods for antimicrobial dilution and disk susceptibility testing of infrequently isolat-ed or fastidious bacteria. 3rd ed. CLSI Guide-line M45. Clinical and Laboratory Stand-ards Institute, Wayne, PA.
14. Wang G, Clark CG, Taylor TM, Pucknell C, Barton C, Price L, Woodward DL, Rodgers FG (2002). Colony multiplex PCR assay for identification and differen-tiation of Campylobacter jejuni, C. coli, C. lari, C. upsaliensis, and C. fetus subsp. fetus. J Clin Microbiol, 40 (12): 4744-7.
15. Zhang A, Song L, Liang H, Gu Y, Zhang C, Liu X, Zhang J, Zhang M (2016). Molec-ular subtyping and erythromycin re-sistance of Campylobacter in China. J Appl Microbiol, 121 (1): 287-93.
16. Gibreel A, Tracz DM, Nonaka L, Ngo TM, Connell SR, Taylor DE (2004). Incidence of antibiotic resistance in Campylobacter je-juni isolated in Alberta, Canada, from 1999 to 2002, with special reference to tet(O)-mediated tetracycline resistance. Antimicrob Agents Chemother, 48(9): 3442-50.
17. Zirnstein G, Li Y, Swaminathan B, Angulo F (1999). Ciprofloxacin resistance in Cam-pylobacter jejuni isolates: detection of gyrA resistance mutations by mismatch ampli-fication mutation assay PCR and DNA sequence analysis. J Clin Microbiol, 37 (10): 3276-80.
18. Zirnstein G, Helsel L, Li Y, Swaminathan B, Besser J (2000). Characterization of gyrA mutations associated with fluoroquino-lone resistance in Campylobacter coli by DNA sequence analysis and MAMA PCR. FEMS Microbiol Lett, 190 (1): 1-7.
19. Alonso R, Mateo E, Churruca E, Martinez I, Girbau C, Fernández-Astorga A (2005). MAMA-PCR assay for the detection of point mutations associated with high-level erythromycin resistance in Campylo-bacter jejuni and Campylobacter coli strains. J Microbiol Methods, 63 (1): 99-103.
20. Alfredson DA, Korolik V (2007). Antibiotic resistance and resistance mechanisms in Campylobacter jejuni and Campylobacter coli. FEMS Microbiol Lett, 277 (2): 123-32.
21. Jonaidi-Jafari N, Khamesipour F, Ranjbar R, Kheiri R (2016). Prevalence and antimi-crobial resistance of Campylobacter species isolated from the avian eggs. Food Con-trol, 70: 35-40.
22. Fitzgerald C (2015). Campylobacter. Clin Lab Med, 35(2): 289-98.
23. Patrick ME, Henao OL, Robinson T, Geis-sler AL, Cronquist A, Hanna S, Hurd S, Medalla F, Pruckler J, Mahon BE (2018). Features of illnesses caused by five spe-cies of Campylobacter, Foodborne Dis-eases Active Surveillance Network (FoodNet) - 2010-2015. Epidemiol Infect, 146(1): 1-10.
24. Ciftci N, Turk-Dagi H, Tuncer I (2019). Prevalence and antimicrobial susceptibility of Campylobacter and Salmonella species in patients with acute gastroenteritis. Klimik Derg, 32 (2): 127-31.
25. Ranjbar R, Babazadeh D, Jonaidi-Jafari N (2017). Prevalence of Campylobacter jejuni in adult patients with inflammatory bacterial diarrhea, East Azerbaijan, Iran. Acta Med Mediterr, 33: 901.
26. Zhang L, Li Y, Shao Y, et al (2020). Molecu-lar characterization and antibiotic resistant profiles of Campylobacter species isolated from poultry and diarrheal patients in Southeastern China 2017–2019. Front Mi-crobiol, 11:1244.
27. Sproston EL, Wimalarathna HML, Shep-pard SK (2018). Trends in fluoroquino-lone resistance in Campylobacter. Microb Ge-nom, 4 (8): e000198.
28. Smith JL, Fratamico PM (2010). Fluoroquin-olone resistance in Campylobacter. J Food Prot, 73 (6): 1141-52.
29. Elhadidy M, Ali MM, El-Shibiny A, Miller WG, Elkhatib WF, Botteldoorn N, Dier-ick K. (2020). Antimicrobial resistance patterns and molecular resistance markers of Campylobacter jejuni isolates from human diarrheal cases. PLoS One, 15 (1): e0227833.
30. Kim JS, Lee MY, Kim SJ, Jeon S-E, Cha I, Hong S, Chung GT, Huh M-J, Kang Y-H, Yoo C-K, Kim J (2016). High-level ciprofloxacin-resistant Campylobacter jejuni isolates circulating in humans and animals in Incheon, Republic of Korea. Zoonoses Public Health, 63 (7): 545-54.
31. Kayman T, Abay S, Hızlısoy H (2013). Iden-tification of Campylobacter spp. isolates with phenotypic methods and multiplex polymerase chain reaction and their anti-biotic susceptibilities. Mikrobiyol Bul, 47 (2): 230-9.
32. Kayman T, Abay S, Aydin F, Şahin O (2019). Antibiotic resistance of Campylo-bacter jejuni isolates recovered from hu-mans with diarrhoea in Turkey. J Med Mi-crobiol, 68 (2): 136-42.
33. Zhou J, Zhang M, Yang W, Fang Y, Wang G, Hou F (2016). A seventeen-year ob-servation of the antimicrobial susceptibil-ity of clinical Campylobacter jejuni and the molecular mechanisms of erythromycin-resistant isolates in Beijing, China. Int J Infect Dis, 42: 28-33.
34. Carev M, Kovačić A, Novak A, Tonkić M, Jerončić A (2017). Campylobacter jejuni strains coresistant to tetracycline and ciprofloxacin in patients with gastroenter-itis in Croatia. Infect Dis (Lond), 49 (4): 268-76.
35. Chen J, Sun XT, Zeng Z, Yu YY (2011). Campylobacter enteritis in adult patients with acute diarrhea from 2005 to 2009 in Beijing, China. Chin Med J (Engl), 124 (10): 1508-12.
36. Shen Z, Wang Y, Zhang Q, Shen J (2018). Antimicrobial resistance in Campylobacter spp. Microbiol Spectr, 6 (2).
37. García‑Fernández A, Dionisi AM, Arena S, Iglesias‑Torrens Y, Carattoli A, Luzzi I (2018). Human campylobacteriosis in Ita-ly: Emergence of multi‑drug resistance to ciprofloxacin, tetracycline, and erythro-mycin. Front Microbiol, 9: 1906.
38. Yılmaz AT, Tuğrul HM (2005). Investigation on Campylobacter species as diarrheogenic agents in Edirne, Turkey, and their anti-microbial susceptibility patterns. Turkish J Infect, 19: 53‑9.
39. Eryildiz C, Tabakcioglu K, Kuyucuklu G, Sakru N (2020). Investigation of antimi-crobial resistance and virulence genes of Campylobacter isolates from patients in a tertiary hospital in Edirne, Turkey. Indian J Med Microbiol, 38 (2): 157-61.
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| Issue | Vol 51 No 3 (2022) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijph.v51i3.8933 | |
| Keywords | ||
| Campylobacter Antimicrobial resistance Resistance genes | ||
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How to Cite
1.
Eryıldız C, Sakru N, Kuyucuklu G. Investigation of Antimicrobial Susceptibilities and Resistance Genes of Campylobacter Isolates from Patients in Edirne, Turkey. Iran J Public Health. 2022;51(3):569-577.
