Research Article
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Identification of Anaerobic Bacteria Isolated From Clinical Samples and Determination of Antibiotic Resistance Profiles

Year 2023, Volume: 7 Issue: 3, 157 - 165, 25.10.2023
https://doi.org/10.34084/bshr.1352333

Abstract

Objective: In this study, it was aimed to identify anaerobic bacteria isolated from various clinical samples, and to determine their antibiotic resistance by gradient method (E-test).
Methods: The study was carried out between January 15 and November 1, 2021. The 213 of 863 samples were included in the study. Anaerobic strains were isolated by conventional methods and identified by an automated system. Antimicrobial susceptibility was determined by the gradient method according to the Clinical and Laboratory Standards Institute (CLSI) criteria.
Results: Anaerobic bacteria were detected in 10.3% of the samples, aerobic/facultative anaerobic bacteria were detected in 34.8%, while growth was not observed in 54.9% of the samples. The 77% of the samples were abscess. The 72.7% of anaerobic bacteria were Gram positive bacteria, and 27.3% were Gram negative bacteria. The most common species were; Cutibacterium (22.7%), Actinomyces (18.3%), Prevotella (13.7%), Bacteroides (9.1%), Anaerococcus (9.1%), Clostridium species (9.1%). The antibiotic susceptibilities of all anaerobic bacteria were as following; moxifloxacin (95.5%), piperacillin-tazobactam (95.5%), amoxicillin-clavulanic acid (95.5%), cefoxitin (90.9%), meropenem (90.9%), clindamycin (77.3%), ampicillin (59.1%), and metronidazole (22.7%), respectively. The susceptibility rates of gram positive bacilli were 91.7% for ampicillin, amoxicillin-clavulanic acid, piperacillin-tazobactam, cefoxitin, moxifloxacin, meropenem, and 75% for clindamycin. In Gram positive cocci, susceptibility to ampicillin was 50%, susceptibility to amoxicillin-clavulanic acid, piperacillin-tazobactam, cefoxitin, clindamycin, moxifloxacin were 100%, and to meropenem was 75%. The susceptibility rates for Gram-negative bacilli were 0.0% for ampicillin, 100% for amoxicillin-clavulanic acid, piperacillin-tazobactam, moxifloxacin, meropenem, 83.3% for metronidazole, 66.7% for cefoxitin, and 50% for clindamycin.
Conclusion: In our study, it was observed that the sensitivity rates for especially, metronidazole and ampicillin were low among anaerobic bacteria. The resistance profile of many anaerobic bacteria has changed significantly over the past decade, making the antimicrobial susceptibility of anaerobic bacteria unpredictable. For this reason, revealing and documenting local data on this subject at regular intervals will constitute an important reference for both empirical treatment, public health, and surveillance studies.

Ethical Statement

Çukurova Üniversitesi Tıp Fakültesi Girişimsel Olmayan Klinik Araştırmalar Etik Kurulu’ndan onay alınmıştır (No: 104, Tarih: 02.10.2020).

Supporting Institution

Bu çalışma Tıpta Uzmanlık Tezi olarak Çukurova Üniversitesi Bilimsel Araştırma Projeleri tarafından TTU2020-13333 No’lu proje ile desteklenmiştir.

Project Number

TTU2020-13333

Thanks

-

References

  • REFERENCES 1. Sood A, Ray P, Angrup A. Antimicrobial susceptibility testing of anaerobic bacteria: In routine and research. Anaerobe. 2022; 75: 102559. doi: 10.1016/j.anaerobe.2022.102559.
  • 2. Winn WC, Koneman EW, Allen SD, et al. The Anaerobic Bacteria (6. Baskı). Konemans Color Atlas and Textbook of Diagnostic Microbiology. Philadelphia: Lippincott Williams & Wilkins Publication, 2006: 877-944.
  • 3. Nagy LE, Boyanova, Justesen US. How to isolate, identify and determine antimicrobial susceptibility of anaerobic bacteria in routine laboratories. Clin Microbiol Infect. 2018; 24(11): 1139-1148. doi: 0.1016/j.cmi.2018.02.008.
  • 4. Hentges DJ. Anaerobes as normal flora. Anaerobic infections in humans, Finegold SM, George WL (Eds), San Diego, Elsevier Publications, 2012; p. 37e53.
  • 5. Boyanova L, Kolarov R, Mitov I. Recent evolution of antibiotic resistance in the anaerobes as compared to previous decades. Anaerobe. 2015; 31: 4e10. doi: 10.1016/j.anaerobe.2014.05.004.
  • 6. Clinical and Laboratory Standards Institute. Methods for antimicrobial susceptibility testing of anaerobic bacteria (9th Ed.), Document M11eA8, Wayne, PA, USA, 2019.
  • 7. Byun JH, Kim M, Lee Y, et al. Antimicrobial Susceptibility Patterns of Anaerobic Bacterial Clinical Isolates From 2014 to 2016, Including Recently Named or Renamed Species. Ann Lab Med. 2019; 39: 190-199. doi: 10.3343/alm.2019.39.2.190.
  • 8. Clinical and Laboratory Standards Institute (2012, January). Performance Standards for Antimicrobial Susceptibility Testing. Twenty-Second Informational Supplement, Document M100-S22, Clinical and Laboratory Standards Institute, Wayne PA, USA. https://m.ibric.org/miniboard/down.php?Board=exp_qna&filename=CLSI%20-%20M100%20S22E.pdf&id=531983&fidx=1 adresinden 30 Haziran 2023’de erişildi.
  • 9. Brook I. Spectrum and treatment of anaerobic infections. J Infect Chemother. 2016; 22: 1e13. doi: 10.1016/j.jiac.2015.10.010.
  • 10. Shah NB, Tande AJ, Patel R, et al. Anaerobic prosthetic joint infection. Anaerobe. 2015; 36: 1-8. doi:10.1016/j.anaerobe.2015.08.003.
  • 11. Gajdács M, Spengler G, Urbán E. Identification and Antimicrobial Susceptibility Testing of Anaerobic Bacteria: Rubik's Cube of Clinical Microbiology?. Antibiotics (Basel). 2017; 6(4): 25. doi:10.3390/antibiotics6040025.
  • 12. Boyanova L, Kolarov R, Mitov I. Recent evolution of antibiotic resistance in the anaerobes as compared to previous decades. Anaerobe. 2015; 31: 4-10. doi:10.1016/j.anaerobe.2014.05.004.
  • 13. Ananth-Shenoy P, Vishwanath S, Targain R, et al. Anaerobic infections in surgical wards: A two year study. Iran J Microbiol. 2016; 8: 181–186.
  • 14. Byun JH, Kim M, Lee Y, et al. Antimicrobial Susceptibility Patterns of Anaerobic Bacterial Clinical Isolates From 2014 to 2016, Including Recently Named or Renamed Species. Ann Lab Med. 2019; 39(2): 190-199. doi:10.3343/alm.2019.39.2.190.
  • 15. Demir C, Keşli R. Çeşitli klinik örneklerden izole edilen Gram-negatif anaerop basillerin tiplendirilmesi ve antibiyotik direnç profillerinin E-test yöntemi ile belirlenmesi [Identification of anaerobic Gram-negative bacilli isolated from various clinical specimens and determination of antibiotic resistance profiles with E-test methods]. Microbiyol Bul. 2018; 52(1): 72-79. doi:10.5578/mb.66175.
  • 16. Özcan N, Saat N, Atmaca N. Klinik örneklerden soyutlanan anaerop bakterilerin in vitro antibiyotik duyarlılıkları. Flora. 2020; 25(2): 245-55. doi: 10.5578/flora.68705.
  • 17. Doğan M, Baysal B. Çeşitli klinik örneklerden izole edilen anaerop bakterilerin tanımlanması ve antibiyotik duyarlılıklarının belirlenmesi. Microbiyol Bul. 2010; 44: 211-219.
  • 18. Uysal BE, Çelik C, Alan Ç, et al. Klinik örneklerden izole edilen anaerobik bakteriler: yedi yıllık değerlendirme. Cumhuriyet Tıp Derg. 2014;36:327-31. doi: 10.7197/cmj.v36i3.5000034023.
  • 19. Özcan N, Bacalan F, Çakır F, et al. Identification and antimicrobial susceptibility testing of anaerobic bacteria isolated from clinical samples. J Bacteriol Mycol Open Access. 2020; 8(1): 29-32. doi: 10.15406/jbmoa.2020.08.00269.
  • 20. Boyanova L, Kolarov R, Gergova G, et al. Anaerobic bacteria in 118 patients with deep-space head and neck infections from the University Hospital of Maxillofacial Surgery, Sofia, Bulgaria. J Med Microbiol. 2006; 55(Pt12): 1759-1760. doi:10.1099/jmm.0.46512-0.
  • 21. Cobo F, Guillot V, Navarro-Marí JM. Breast Abscesses Caused by Anaerobic Microorganisms: Clinical and Microbiological Characteristics. Antibiotics (Basel). 2020; 9(6): 341. doi:10.3390/antibiotics9060341.
  • 22. Byun JH, Kim M, Lee Y, et al. Antimicrobial Susceptibility Patterns of Anaerobic Bacterial Clinical Isolates From 2014 to 2016, Including Recently Named or Renamed Species. Ann Lab Med. 2019; 39(2): 190-199. doi:10.3343/alm.2019.39.2.190.
  • 23. Wybo I, Van den Bossche D, Soetens O, et al. Fourth Belgian multicentre survey of antibiotic susceptibility of anaerobic bacteria. J Antimicrob Chemother. 2014;69(1):155-161. doi:10.1093/jac/dkt344.
  • 24. Badr MT, Blümel B, Baumgartner S, et al. Antimicrobial Susceptibility Patterns and Wild-Type MIC Distributions of Anaerobic Bacteria at a German University Hospital: A Five-Year Retrospective Study (2015-2019). Antibiotics (Basel). 2020; 9(11): 823. doi:10.3390/antibiotics9110823.
  • 25. Ishak N, Abdul Wahab Z, Amin Nordin S, et al. Susceptibility patterns of anaerobes isolated from clinical specimens in tertiary Hospital, Malaysia. Malays J Pathol. 2020; 42(2): 245-252.
  • 26. Naidoo S, Perovic O, Richards GA, Duse AG. Clinically significant anaerobic bacteria isolated from patients in a South African academic hospital: antimicrobial susceptibility testing. S Afr Med J. 2011;101(10):732-734.
  • 27. Bozkurt H, Güdücüoğlu H, Bayram Y, et al. Klinik örneklerden izole edilen anaerob bakteriler ve antibiyotik duyarlılıkları. Van Tıp Derg. 2004; 11: 85-91.
  • 28. Jeverica S, Kolenc U, Mueller-Premru M, et al. Evaluation of the routine antimicrobial susceptibility testing results of clinically significant anaerobic bacteria in a Slovenian tertiary-care hospital in 2015. Anaerobe. 2017; 47: 64-69. doi:10.1016/j.anaerobe.2017.04.007.
  • 29. Katsandri A, Avlamis A, Pantazatou A, et al. In vitro activities of tigecycline against recently isolated Gram-negative anaerobic bacteria in Greece, including metronidazole-resistant strains. Diagn Microbiol Infect Dis. 2006; 55(3): 231-236. doi:10.1016/j.diagmicrobio.2006.01.022.
  • 30. Akhi MT, Ghotaslou R, Beheshtirouy S, et al. Antibiotic Susceptibility Pattern of Aerobic and Anaerobic Bacteria Isolated From Surgical Site Infection of Hospitalized Patients. Jundishapur J Microbiol. 2015; 8(7): e20309. doi:10.5812/jjm.20309v2.
  • 31. Lee Y, Park Y, Kim MS, et al. Antimicrobial susceptibility patterns for recent clinical isolates of anaerobic bacteria in South Korea. Antimicrob Agents Chemother. 2010; 54(9): 3993-3997. doi:10.1128/AAC.00481-10

Klinik Örneklerden İzole Edilen Anaerop Bakterilerin Tiplendirilmesi ve Antibiyotik Direnç Profillerinin Belirlenmesi

Year 2023, Volume: 7 Issue: 3, 157 - 165, 25.10.2023
https://doi.org/10.34084/bshr.1352333

Abstract

ÖZ
Amaç: Bu çalışmada, çeşitli klinik örneklerden izole edilen anaerop bakterilerin tanımlanması ve gradiyent yöntemi (E-test) ile antibiyotik dirençlerinin belirlenmesi amaçlandı.
Yöntem: Çalışma, 15 Ocak-1 Kasım 2021 tarihlerinde gerçekleştirildi. 863 örneğin 213’ü çalışmaya dahil edildi. Anaerop suşlar konvansiyonel yöntemlerle izole edildi ve otomatize sistemle tanımlandı. Antibiyotik duyarlılıkları Clinical and Laboratory Standards Institute (CLSI) kriterlerine göre gradiyent yöntemi ile belirlendi.
Bulgular: Örneklerin %10,3’ünde anaerop bakteri, %34,8’inde aerop/fakültatif anaerop bakteri tespit edilirken, %54,9’unda üreme görülmedi. Örneklerin %77’si apse materyaliydi. Anaerop bakterilerin %72,7’si Gram pozitif bakteri, %27,3’ü Gram negatif basildi. En yaygın türler; Cutibacterium (%22,7), Actinomyces (%18,3), Prevotella (%13,7), Bacteroides (%9,1), Anaerococcus (%9,1), Clostridium türleri (%9,1) idi. Sırasıyla, tüm anaerop bakterilerin antibiyotik duyarlılıkları; moksifloksasin (%95,5), piperasilin-tazobaktam (%95,5), amoksisilin-klavulonik asit (%95,5), sefoksitin (%90,9), meropenem (%90,9) klindamisin (%77,3), ampisilin (%59,1) ve metronidazol (%22,7) idi. Gram pozitif basillerin duyarlılık oranları ampisilin, amoksisillin-klavulanik asit, piperasilin-tazobaktam, sefoksitine, moksifloksasin, meropenem için %91,7, klindamisin için %75’di. Gram pozitif koklarda ampisiline duyarlılık %50, amoksisillin-klavulanik asit, piperasilin-tazobaktam, sefoksitin, klindamisin, moksifloksasine duyarlılık %100 iken, meropeneme duyarlılık %75’di. Gram negatif basillerde duyarlılık oranları ampisilin %0,0, amoksisillin-klavulanik asit, piperasilin-tazobaktam, moksifloksasin, meropenem için %100, metronidazol %83,3, sefoksitin için %66,7 ve klindamisin için %50 idi.
Sonuç: Çalışmamızda anaerop bakterilerde özellikle metronidazol ve ampisilin için duyarlılık oranlarının düşük olduğu gözlemlendi. Birçok anaerop bakterinin direnç profilinin, son on yılda önemli ölçüde değişmesi, anaerop bakterilerin antimikrobiyal duyarlılıklarını fazla tahmin edilemez hale getirmiştir. Bu sebeple bu konudaki lokal verilerin belli aralıklarla ortaya çıkarılması ve dökümante edilmesi hem ampirik tedavinin şekillenmesi, hem halk sağlığı, hem de surveyans çalışmaları için önemli bir referans oluşturacaktır.

Project Number

TTU2020-13333

References

  • REFERENCES 1. Sood A, Ray P, Angrup A. Antimicrobial susceptibility testing of anaerobic bacteria: In routine and research. Anaerobe. 2022; 75: 102559. doi: 10.1016/j.anaerobe.2022.102559.
  • 2. Winn WC, Koneman EW, Allen SD, et al. The Anaerobic Bacteria (6. Baskı). Konemans Color Atlas and Textbook of Diagnostic Microbiology. Philadelphia: Lippincott Williams & Wilkins Publication, 2006: 877-944.
  • 3. Nagy LE, Boyanova, Justesen US. How to isolate, identify and determine antimicrobial susceptibility of anaerobic bacteria in routine laboratories. Clin Microbiol Infect. 2018; 24(11): 1139-1148. doi: 0.1016/j.cmi.2018.02.008.
  • 4. Hentges DJ. Anaerobes as normal flora. Anaerobic infections in humans, Finegold SM, George WL (Eds), San Diego, Elsevier Publications, 2012; p. 37e53.
  • 5. Boyanova L, Kolarov R, Mitov I. Recent evolution of antibiotic resistance in the anaerobes as compared to previous decades. Anaerobe. 2015; 31: 4e10. doi: 10.1016/j.anaerobe.2014.05.004.
  • 6. Clinical and Laboratory Standards Institute. Methods for antimicrobial susceptibility testing of anaerobic bacteria (9th Ed.), Document M11eA8, Wayne, PA, USA, 2019.
  • 7. Byun JH, Kim M, Lee Y, et al. Antimicrobial Susceptibility Patterns of Anaerobic Bacterial Clinical Isolates From 2014 to 2016, Including Recently Named or Renamed Species. Ann Lab Med. 2019; 39: 190-199. doi: 10.3343/alm.2019.39.2.190.
  • 8. Clinical and Laboratory Standards Institute (2012, January). Performance Standards for Antimicrobial Susceptibility Testing. Twenty-Second Informational Supplement, Document M100-S22, Clinical and Laboratory Standards Institute, Wayne PA, USA. https://m.ibric.org/miniboard/down.php?Board=exp_qna&filename=CLSI%20-%20M100%20S22E.pdf&id=531983&fidx=1 adresinden 30 Haziran 2023’de erişildi.
  • 9. Brook I. Spectrum and treatment of anaerobic infections. J Infect Chemother. 2016; 22: 1e13. doi: 10.1016/j.jiac.2015.10.010.
  • 10. Shah NB, Tande AJ, Patel R, et al. Anaerobic prosthetic joint infection. Anaerobe. 2015; 36: 1-8. doi:10.1016/j.anaerobe.2015.08.003.
  • 11. Gajdács M, Spengler G, Urbán E. Identification and Antimicrobial Susceptibility Testing of Anaerobic Bacteria: Rubik's Cube of Clinical Microbiology?. Antibiotics (Basel). 2017; 6(4): 25. doi:10.3390/antibiotics6040025.
  • 12. Boyanova L, Kolarov R, Mitov I. Recent evolution of antibiotic resistance in the anaerobes as compared to previous decades. Anaerobe. 2015; 31: 4-10. doi:10.1016/j.anaerobe.2014.05.004.
  • 13. Ananth-Shenoy P, Vishwanath S, Targain R, et al. Anaerobic infections in surgical wards: A two year study. Iran J Microbiol. 2016; 8: 181–186.
  • 14. Byun JH, Kim M, Lee Y, et al. Antimicrobial Susceptibility Patterns of Anaerobic Bacterial Clinical Isolates From 2014 to 2016, Including Recently Named or Renamed Species. Ann Lab Med. 2019; 39(2): 190-199. doi:10.3343/alm.2019.39.2.190.
  • 15. Demir C, Keşli R. Çeşitli klinik örneklerden izole edilen Gram-negatif anaerop basillerin tiplendirilmesi ve antibiyotik direnç profillerinin E-test yöntemi ile belirlenmesi [Identification of anaerobic Gram-negative bacilli isolated from various clinical specimens and determination of antibiotic resistance profiles with E-test methods]. Microbiyol Bul. 2018; 52(1): 72-79. doi:10.5578/mb.66175.
  • 16. Özcan N, Saat N, Atmaca N. Klinik örneklerden soyutlanan anaerop bakterilerin in vitro antibiyotik duyarlılıkları. Flora. 2020; 25(2): 245-55. doi: 10.5578/flora.68705.
  • 17. Doğan M, Baysal B. Çeşitli klinik örneklerden izole edilen anaerop bakterilerin tanımlanması ve antibiyotik duyarlılıklarının belirlenmesi. Microbiyol Bul. 2010; 44: 211-219.
  • 18. Uysal BE, Çelik C, Alan Ç, et al. Klinik örneklerden izole edilen anaerobik bakteriler: yedi yıllık değerlendirme. Cumhuriyet Tıp Derg. 2014;36:327-31. doi: 10.7197/cmj.v36i3.5000034023.
  • 19. Özcan N, Bacalan F, Çakır F, et al. Identification and antimicrobial susceptibility testing of anaerobic bacteria isolated from clinical samples. J Bacteriol Mycol Open Access. 2020; 8(1): 29-32. doi: 10.15406/jbmoa.2020.08.00269.
  • 20. Boyanova L, Kolarov R, Gergova G, et al. Anaerobic bacteria in 118 patients with deep-space head and neck infections from the University Hospital of Maxillofacial Surgery, Sofia, Bulgaria. J Med Microbiol. 2006; 55(Pt12): 1759-1760. doi:10.1099/jmm.0.46512-0.
  • 21. Cobo F, Guillot V, Navarro-Marí JM. Breast Abscesses Caused by Anaerobic Microorganisms: Clinical and Microbiological Characteristics. Antibiotics (Basel). 2020; 9(6): 341. doi:10.3390/antibiotics9060341.
  • 22. Byun JH, Kim M, Lee Y, et al. Antimicrobial Susceptibility Patterns of Anaerobic Bacterial Clinical Isolates From 2014 to 2016, Including Recently Named or Renamed Species. Ann Lab Med. 2019; 39(2): 190-199. doi:10.3343/alm.2019.39.2.190.
  • 23. Wybo I, Van den Bossche D, Soetens O, et al. Fourth Belgian multicentre survey of antibiotic susceptibility of anaerobic bacteria. J Antimicrob Chemother. 2014;69(1):155-161. doi:10.1093/jac/dkt344.
  • 24. Badr MT, Blümel B, Baumgartner S, et al. Antimicrobial Susceptibility Patterns and Wild-Type MIC Distributions of Anaerobic Bacteria at a German University Hospital: A Five-Year Retrospective Study (2015-2019). Antibiotics (Basel). 2020; 9(11): 823. doi:10.3390/antibiotics9110823.
  • 25. Ishak N, Abdul Wahab Z, Amin Nordin S, et al. Susceptibility patterns of anaerobes isolated from clinical specimens in tertiary Hospital, Malaysia. Malays J Pathol. 2020; 42(2): 245-252.
  • 26. Naidoo S, Perovic O, Richards GA, Duse AG. Clinically significant anaerobic bacteria isolated from patients in a South African academic hospital: antimicrobial susceptibility testing. S Afr Med J. 2011;101(10):732-734.
  • 27. Bozkurt H, Güdücüoğlu H, Bayram Y, et al. Klinik örneklerden izole edilen anaerob bakteriler ve antibiyotik duyarlılıkları. Van Tıp Derg. 2004; 11: 85-91.
  • 28. Jeverica S, Kolenc U, Mueller-Premru M, et al. Evaluation of the routine antimicrobial susceptibility testing results of clinically significant anaerobic bacteria in a Slovenian tertiary-care hospital in 2015. Anaerobe. 2017; 47: 64-69. doi:10.1016/j.anaerobe.2017.04.007.
  • 29. Katsandri A, Avlamis A, Pantazatou A, et al. In vitro activities of tigecycline against recently isolated Gram-negative anaerobic bacteria in Greece, including metronidazole-resistant strains. Diagn Microbiol Infect Dis. 2006; 55(3): 231-236. doi:10.1016/j.diagmicrobio.2006.01.022.
  • 30. Akhi MT, Ghotaslou R, Beheshtirouy S, et al. Antibiotic Susceptibility Pattern of Aerobic and Anaerobic Bacteria Isolated From Surgical Site Infection of Hospitalized Patients. Jundishapur J Microbiol. 2015; 8(7): e20309. doi:10.5812/jjm.20309v2.
  • 31. Lee Y, Park Y, Kim MS, et al. Antimicrobial susceptibility patterns for recent clinical isolates of anaerobic bacteria in South Korea. Antimicrob Agents Chemother. 2010; 54(9): 3993-3997. doi:10.1128/AAC.00481-10
There are 31 citations in total.

Details

Primary Language English
Subjects Medical Bacteriology, Medical Infection Agents
Journal Section Research Article
Authors

Tünzala Asgarova 0000-0003-4000-5095

Filiz Kibar 0000-0003-2983-2399

Hatice Hale Gümüş 0000-0001-9071-9606

Project Number TTU2020-13333
Early Pub Date October 27, 2023
Publication Date October 25, 2023
Acceptance Date September 16, 2023
Published in Issue Year 2023 Volume: 7 Issue: 3

Cite

AMA Asgarova T, Kibar F, Gümüş HH. Identification of Anaerobic Bacteria Isolated From Clinical Samples and Determination of Antibiotic Resistance Profiles. J Biotechnol and Strategic Health Res. October 2023;7(3):157-165. doi:10.34084/bshr.1352333

Journal of Biotechnology and Strategic Health Research