Exploring the genetic diversity and the association of drug resistance and biofilm production in Acinetobacter baumannii strains isolated from burn wound infections
,
Abstract
Background and Objectives: Acinetobacter baumannii is considered a troublesome cause of infection in burn units, where its capability to form biofilm and resist antibiotics significantly hampers therapeutic success. This study explored the correlations between antimicrobial resistance profiles, biofilm-producing capacity, and genetic diversity of A. baumannii strains from patients with burn wound infection in Isfahan, Iran.
Materials and Methods: Ninety-six isolates were analyzed for antibiotic resistance using the disk diffusion technique and for biofilm formation through the microtiter dish assay. The prevalence of ten biofilm-related genes was investigated using specific primers. Clonal relatedness among bacterial strains was defined by Enterobacterial Repetitive Intergenic Consensus-Polymerase Chain Reaction (ERIC-PCR).
Results: A vast majority of isolates (99%) exhibited resistance to meropenem, ciprofloxacin, ceftriaxone, cefotaxime, piperacillin-tazobactam, and imipenem, qualifying them as extensively drug-resistant (XDR). Twenty-five percent of the strains were strong biofilm formers, while 68% demonstrated moderate or weak biofilm formation. The most commonly identified biofilm-related genes included bfmR (100%), ompA (100%), and bap (99%). A significant association was found between the production of biofilm, resistance to aminoglycosides, and the presence of csuE and bap genes. ERIC-PCR typing showed the presence of 3 clonal types and 7 single types, with biofilm producers predominantly clustering to clonal type 2.
Conclusion: This work highlights a notable prevalence of biofilm-producing XDR A. baumannii in burn patients, underscoring the need for continuous surveillance and enhanced infection control strategies.
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2. Norbury W, Herndon DN, Tanksley J, Jeschke MG, Finnerty CC. Infection in burns. Surg Infect (Larchmt) 2016; 17: 250-255.
3. Zeighami H, Valadkhani F, Shapouri R, Samadi E, Haghi F. Virulence characteristics of multidrug resistant biofilm forming Acinetobacter baumannii isolated from intensive care unit patients. BMC Infect Dis 2019; 19: 629.
4. Amin M, Navidifar T, Shooshtari FS, Rashno M, Savari M, Jahangirmehr F, et al. Association between biofilm formation, structure, and the expression levels of genes related to biofilm formation and biofilm-specific resistance of Acinetobacter baumannii strains isolated from burn infection in Ahvaz, Iran. Infect Drug Resist 2019; 12: 3867-3881.
5. Khoshnood S, Sadeghifard N, Mahdian N, Heidary M, Mahdian S, Mohammadi M, et al. Antimicrobial resistance and biofilm formation capacity among Acinetobacter baumannii strains isolated from patients with burns and ventilator‐associated pneumonia. J Clin Lab Anal 2023; 37(1): e24814.
6. Kim H-J, Kim N-Y, Ko S-Y, Park S-Y, Oh M-H, Shin M-S, et al. Complementary regulation of BfmRS two-component and AbaIR quorum sensing systems to express virulence-associated genes in Acinetobacter baumannii. Int J Mol Sci 2022; 23: 13136.
7. Abd El-Rahman OA, Rasslan F, Hassan SS, Ashour HM, Wasfi R. The RND efflux pump gene expression in the biofilm formation of Acinetobacter baumannii. Antibiotics (Basel) 2023; 12: 419.
8. Mahmoudi H, Shokoohizadeh L, Zare Fahim N, Mohamadi Bardebari A, Moradkhani S, Alikhani MY. Detection of adeABC efllux pump encoding genes and antimicrobial effect of Mentha longifolia and Menthol on MICs of imipenem and ciprofloxacin in clinical isolates of Acinetobacter baumannii. BMC Complement Med Ther 2020; 20: 92.
9. Dolma KG, Khati R, Paul AK, Rahmatullah M, de Lourdes Pereira M, Wilairatana P, et al. Virulence characteristics and emerging therapies for biofilm-forming Acinetobacter baumannii: a review. Biology (Basel) 2022; 11: 1343.
10. Mostafavi NS, Rahimi F, Khashei S. Characterization of clonal groups of antibiotic-resistant biofilm-forming Staphylococcus aureus strains isolated from patients with urinary tract infections in Isfahan, Iran. Arch Clin Infect Dis 2024; 19(4): e143383.
11. Mobarak-Qamsari M, Jenaghi B, Sahebi L, Norouzi-Shadehi M, Salehi M-R, Shakoori-Farahani A, et al. Evaluation of Acinetobacter baumannii, Klebsiella pneumoniae, and Staphylococcus aureus respiratory tract superinfections among patients with COVID-19 at a tertiary-care hospital in Tehran, Iran. Eur J Med Res 2023; 28: 314.
12. Mobarak Qamsari M, Sahebi L, Salehi MR, Labbani-Motlagh Z, Shavandi M, Alijani N, et al. Respiratory bacterial and fungal superinfections during the third surge of the COVID-19 pandemic in Iran. Microb Drug Resist 2023; 29: 104-111.
13. Falah F, Shokoohizadeh L, Adabi M. Molecular identification and genotyping of Acinetobacter baumannii isolated from burn patients by PCR and ERIC-PCR. Scars Burn Heal 2019; 5: 2059513119831369.
14. CLSI (2022). CLSI Publishes CLSI M100—Performance Standards for Antimicrobial Susceptibility Testing, 32nd Edition.
https://clsi.org/about/news/clsi-publishes-m100-performance-standards-for-antimicrobial-susceptibility-testing-32nd-edition/
15. ECDC (2017). European Centre for Disease Prevention and Control. Antimicrobial resistance surveillance in Europe 2015. Annual Report of the European Antimicrobial Resistance Surveillance Network (EARS-Net). https://www.ecdc.europa.eu/en/publications-data/antimicrobial-resistance-surveillance-europe-2015
16. Chen L, Li H, Wen H, Zhao B, Niu Y, Mo Q, et al. Biofilm formation in Acinetobacter baumannii was inhibited by PAβN while it had no association with antibiotic resistance. Microbiologyopen 2020; 9(9): e1063.
17. Khashei S, Fazeli H, Rahimi F, Karbasizadeh V. Antibiotic-potentiating efficacy of Rosmarinus officinalis L. to combat planktonic cells, biofilms, and efflux pump activities of extensively drug-resistant Acinetobacter baumannii clinical strains. Front Pharmacol 2025; 16: 1558611.
18. Al-Shamiri MM, Zhang S, Mi P, Liu Y, Xun M, Yang E, et al. Phenotypic and genotypic characteristics of Acinetobacter baumannii enrolled in the relationship among antibiotic resistance, biofilm formation and motility. Microb Pathog 2021; 155: 104922.
19. Farshadzadeh Z, Hashemi FB, Rahimi S, Pourakbari B, Esmaeili D, Haghighi MA, et al. Wide distribution of carbapenem resistant Acinetobacter baumannii in burns patients in Iran. Front Microbiol 2015; 6: 1146.
20. Maleki A, Kaviar VH, Koupaei M, Haddadi MH, Kalani BS, Valadbeigi H, et al. Molecular typing and antibiotic resistance patterns among clinical isolates of Acinetobacter baumannii recovered from burn patients in Tehran, Iran. Front Microbiol 2022; 13: 994303.
21. Sannathimmappa MB, Nambiar V, Aravindakshan R. Antibiotic resistance pattern of Acinetobacter baumannii strains: a retrospective study from Oman. Saudi J Med Med Sci 2021; 9: 254-260.
22. Al-Kadmy IMS, Ali ANM, Salman IMA, Khazaal SS. Molecular characterization of Acinetobacter baumannii isolated from Iraqi hospital environment. New Microbes New Infect 2017; 21: 51-57.
23. Bello-López E, Rocha-Gracia RDC, Castro-Jaimes S, Cevallos MÁ, Vargas-Cruz M, Verdugo-Yocupicio R, et al. Antibiotic resistance mechanisms in Acinetobacter spp. strains isolated from patients in a paediatric hospital in Mexico. J Glob Antimicrob Resist 2020; 23: 120-129.
24. Hujer KM, Hujer AM, Hulten EA, Bajaksouzian S, Adams JM, Donskey CJ, et al. Analysis of antibiotic resistance genes in multidrug-resistant Acinetobacter sp. isolates from military and civilian patients treated at the Walter Reed Army Medical Center. Antimicrob Agents Chemother 2006; 50: 4114-4123.
25. Mahdian S, Sadeghifard N, Pakzad I, Ghanbari F, Soroush S, Azimi L, et al. Acinetobacter baumannii clonal lineages I and II harboring different carbapenem-hydrolyzing-β-lactamase genes are widespread among hospitalized burn patients in Tehran. J Infect Public Health 2015; 8: 533-542.
26. Thomas RE, Thomas BC. Reducing biofilm infections in burn patients’ wounds and biofilms on surfaces in hospitals, medical facilities and medical equipment to improve burn care: a systematic review. Int J Environ Res Public Health 2021; 18: 13195.
27. Donadu MG, Mazzarello V, Cappuccinelli P, Zanetti S, Madléna M, Nagy ÁL, et al. Relationship between the biofilm-forming capacity and antimicrobial resistance in clinical Acinetobacter baumannii isolates: results from a laboratory-based in vitro study. Microorganisms 2021; 9: 2384.
28. Yang C-H, Su P-W, Moi S-H, Chuang L-Y. Biofilm formation in Acinetobacter baumannii: genotype-phenotype correlation. Molecules 2019; 24: 1849.
29. Hoffman LR, D'Argenio DA, MacCoss MJ, Zhang Z, Jones RA, Miller SI. Aminoglycoside antibiotics induce bacterial biofilm formation. Nature 2005; 436: 1171-1175.
30. Penesyan A, Paulsen IT, Gillings MR, Kjelleberg S, Manefield MJ. Secondary effects of antibiotics on microbial biofilms. Front Microbiol 2020; 11: 2109.
31. López D, Vlamakis H, Kolter R. Biofilms. Cold Spring Harb Perspect Biol 2010; 2: a000398.
32. de la Fuente-Núñez C, Reffuveille F, Fernández L, Hancock RE. Bacterial biofilm development as a multicellular adaptation: antibiotic resistance and new therapeutic strategies. Curr Opin Microbiol 2013; 16: 580-589.
33. Gaddy JA, Actis LA. Regulation of Acinetobacter baumannii biofilm formation. Future Microbiol 2009; 4: 273-278.
34. Tomaras AP, Dorsey CW, Edelmann RE, Actis LA. Attachment to and biofilm formation on abiotic surfaces by Acinetobacter baumannii: involvement of a novel chaperone-usher pili assembly system. Microbiology (Reading) 2003; 149: 3473-3484.
35. Ribeiro EA, Gales AC, Oliveira APS, Coelho DD, Oliveira RA, Pfrimer IAH, et al. Molecular epidemiology and drug resistance of Acinetobacter baumannii isolated from a regional hospital in the Brazilian Amazon region. Rev Soc Bras Med Trop 2020; 54: e20200087.
2. Norbury W, Herndon DN, Tanksley J, Jeschke MG, Finnerty CC. Infection in burns. Surg Infect (Larchmt) 2016; 17: 250-255.
3. Zeighami H, Valadkhani F, Shapouri R, Samadi E, Haghi F. Virulence characteristics of multidrug resistant biofilm forming Acinetobacter baumannii isolated from intensive care unit patients. BMC Infect Dis 2019; 19: 629.
4. Amin M, Navidifar T, Shooshtari FS, Rashno M, Savari M, Jahangirmehr F, et al. Association between biofilm formation, structure, and the expression levels of genes related to biofilm formation and biofilm-specific resistance of Acinetobacter baumannii strains isolated from burn infection in Ahvaz, Iran. Infect Drug Resist 2019; 12: 3867-3881.
5. Khoshnood S, Sadeghifard N, Mahdian N, Heidary M, Mahdian S, Mohammadi M, et al. Antimicrobial resistance and biofilm formation capacity among Acinetobacter baumannii strains isolated from patients with burns and ventilator‐associated pneumonia. J Clin Lab Anal 2023; 37(1): e24814.
6. Kim H-J, Kim N-Y, Ko S-Y, Park S-Y, Oh M-H, Shin M-S, et al. Complementary regulation of BfmRS two-component and AbaIR quorum sensing systems to express virulence-associated genes in Acinetobacter baumannii. Int J Mol Sci 2022; 23: 13136.
7. Abd El-Rahman OA, Rasslan F, Hassan SS, Ashour HM, Wasfi R. The RND efflux pump gene expression in the biofilm formation of Acinetobacter baumannii. Antibiotics (Basel) 2023; 12: 419.
8. Mahmoudi H, Shokoohizadeh L, Zare Fahim N, Mohamadi Bardebari A, Moradkhani S, Alikhani MY. Detection of adeABC efllux pump encoding genes and antimicrobial effect of Mentha longifolia and Menthol on MICs of imipenem and ciprofloxacin in clinical isolates of Acinetobacter baumannii. BMC Complement Med Ther 2020; 20: 92.
9. Dolma KG, Khati R, Paul AK, Rahmatullah M, de Lourdes Pereira M, Wilairatana P, et al. Virulence characteristics and emerging therapies for biofilm-forming Acinetobacter baumannii: a review. Biology (Basel) 2022; 11: 1343.
10. Mostafavi NS, Rahimi F, Khashei S. Characterization of clonal groups of antibiotic-resistant biofilm-forming Staphylococcus aureus strains isolated from patients with urinary tract infections in Isfahan, Iran. Arch Clin Infect Dis 2024; 19(4): e143383.
11. Mobarak-Qamsari M, Jenaghi B, Sahebi L, Norouzi-Shadehi M, Salehi M-R, Shakoori-Farahani A, et al. Evaluation of Acinetobacter baumannii, Klebsiella pneumoniae, and Staphylococcus aureus respiratory tract superinfections among patients with COVID-19 at a tertiary-care hospital in Tehran, Iran. Eur J Med Res 2023; 28: 314.
12. Mobarak Qamsari M, Sahebi L, Salehi MR, Labbani-Motlagh Z, Shavandi M, Alijani N, et al. Respiratory bacterial and fungal superinfections during the third surge of the COVID-19 pandemic in Iran. Microb Drug Resist 2023; 29: 104-111.
13. Falah F, Shokoohizadeh L, Adabi M. Molecular identification and genotyping of Acinetobacter baumannii isolated from burn patients by PCR and ERIC-PCR. Scars Burn Heal 2019; 5: 2059513119831369.
14. CLSI (2022). CLSI Publishes CLSI M100—Performance Standards for Antimicrobial Susceptibility Testing, 32nd Edition.
https://clsi.org/about/news/clsi-publishes-m100-performance-standards-for-antimicrobial-susceptibility-testing-32nd-edition/
15. ECDC (2017). European Centre for Disease Prevention and Control. Antimicrobial resistance surveillance in Europe 2015. Annual Report of the European Antimicrobial Resistance Surveillance Network (EARS-Net). https://www.ecdc.europa.eu/en/publications-data/antimicrobial-resistance-surveillance-europe-2015
16. Chen L, Li H, Wen H, Zhao B, Niu Y, Mo Q, et al. Biofilm formation in Acinetobacter baumannii was inhibited by PAβN while it had no association with antibiotic resistance. Microbiologyopen 2020; 9(9): e1063.
17. Khashei S, Fazeli H, Rahimi F, Karbasizadeh V. Antibiotic-potentiating efficacy of Rosmarinus officinalis L. to combat planktonic cells, biofilms, and efflux pump activities of extensively drug-resistant Acinetobacter baumannii clinical strains. Front Pharmacol 2025; 16: 1558611.
18. Al-Shamiri MM, Zhang S, Mi P, Liu Y, Xun M, Yang E, et al. Phenotypic and genotypic characteristics of Acinetobacter baumannii enrolled in the relationship among antibiotic resistance, biofilm formation and motility. Microb Pathog 2021; 155: 104922.
19. Farshadzadeh Z, Hashemi FB, Rahimi S, Pourakbari B, Esmaeili D, Haghighi MA, et al. Wide distribution of carbapenem resistant Acinetobacter baumannii in burns patients in Iran. Front Microbiol 2015; 6: 1146.
20. Maleki A, Kaviar VH, Koupaei M, Haddadi MH, Kalani BS, Valadbeigi H, et al. Molecular typing and antibiotic resistance patterns among clinical isolates of Acinetobacter baumannii recovered from burn patients in Tehran, Iran. Front Microbiol 2022; 13: 994303.
21. Sannathimmappa MB, Nambiar V, Aravindakshan R. Antibiotic resistance pattern of Acinetobacter baumannii strains: a retrospective study from Oman. Saudi J Med Med Sci 2021; 9: 254-260.
22. Al-Kadmy IMS, Ali ANM, Salman IMA, Khazaal SS. Molecular characterization of Acinetobacter baumannii isolated from Iraqi hospital environment. New Microbes New Infect 2017; 21: 51-57.
23. Bello-López E, Rocha-Gracia RDC, Castro-Jaimes S, Cevallos MÁ, Vargas-Cruz M, Verdugo-Yocupicio R, et al. Antibiotic resistance mechanisms in Acinetobacter spp. strains isolated from patients in a paediatric hospital in Mexico. J Glob Antimicrob Resist 2020; 23: 120-129.
24. Hujer KM, Hujer AM, Hulten EA, Bajaksouzian S, Adams JM, Donskey CJ, et al. Analysis of antibiotic resistance genes in multidrug-resistant Acinetobacter sp. isolates from military and civilian patients treated at the Walter Reed Army Medical Center. Antimicrob Agents Chemother 2006; 50: 4114-4123.
25. Mahdian S, Sadeghifard N, Pakzad I, Ghanbari F, Soroush S, Azimi L, et al. Acinetobacter baumannii clonal lineages I and II harboring different carbapenem-hydrolyzing-β-lactamase genes are widespread among hospitalized burn patients in Tehran. J Infect Public Health 2015; 8: 533-542.
26. Thomas RE, Thomas BC. Reducing biofilm infections in burn patients’ wounds and biofilms on surfaces in hospitals, medical facilities and medical equipment to improve burn care: a systematic review. Int J Environ Res Public Health 2021; 18: 13195.
27. Donadu MG, Mazzarello V, Cappuccinelli P, Zanetti S, Madléna M, Nagy ÁL, et al. Relationship between the biofilm-forming capacity and antimicrobial resistance in clinical Acinetobacter baumannii isolates: results from a laboratory-based in vitro study. Microorganisms 2021; 9: 2384.
28. Yang C-H, Su P-W, Moi S-H, Chuang L-Y. Biofilm formation in Acinetobacter baumannii: genotype-phenotype correlation. Molecules 2019; 24: 1849.
29. Hoffman LR, D'Argenio DA, MacCoss MJ, Zhang Z, Jones RA, Miller SI. Aminoglycoside antibiotics induce bacterial biofilm formation. Nature 2005; 436: 1171-1175.
30. Penesyan A, Paulsen IT, Gillings MR, Kjelleberg S, Manefield MJ. Secondary effects of antibiotics on microbial biofilms. Front Microbiol 2020; 11: 2109.
31. López D, Vlamakis H, Kolter R. Biofilms. Cold Spring Harb Perspect Biol 2010; 2: a000398.
32. de la Fuente-Núñez C, Reffuveille F, Fernández L, Hancock RE. Bacterial biofilm development as a multicellular adaptation: antibiotic resistance and new therapeutic strategies. Curr Opin Microbiol 2013; 16: 580-589.
33. Gaddy JA, Actis LA. Regulation of Acinetobacter baumannii biofilm formation. Future Microbiol 2009; 4: 273-278.
34. Tomaras AP, Dorsey CW, Edelmann RE, Actis LA. Attachment to and biofilm formation on abiotic surfaces by Acinetobacter baumannii: involvement of a novel chaperone-usher pili assembly system. Microbiology (Reading) 2003; 149: 3473-3484.
35. Ribeiro EA, Gales AC, Oliveira APS, Coelho DD, Oliveira RA, Pfrimer IAH, et al. Molecular epidemiology and drug resistance of Acinetobacter baumannii isolated from a regional hospital in the Brazilian Amazon region. Rev Soc Bras Med Trop 2020; 54: e20200087.
| Files | ||
| Issue | Vol 17 No 4 (2025) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v17i4.19228 | |
| Keywords | ||
| Burns Acinetobacter baumannii Drug resistance Biofilm Molecular typing | ||
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How to Cite
1.
Khashei S, Fazeli H, Rahimi F, Karbasizade V. Exploring the genetic diversity and the association of drug resistance and biofilm production in Acinetobacter baumannii strains isolated from burn wound infections. Iran J Microbiol. 2025;17(4):549-558.
