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Assessment of biofilm formation among clinical isolates of Acinetobacter baumannii in burn wounds in the west of Iran
Corresponding Author(s) : Amirhoushang Alvandi
Cellular and Molecular Biology,
Vol. 64 No. 15: Issue 15
Abstract
Burn wound infection by A. baumannii is one of the predominant cause of mortality worldwide. The present investigation aimed at determination of antimicrobial resistance profile and expression of the biofilm-related genes in A. baumannii isolated from hospitalized patients with burn wound infection in Kermanshah hospitals. Sixty four isolates of A. baumannii were recovered from burn wound of hospitalized patients at hospitals in Kermanshah. The antimicrobial susceptibility testing (AST) was performed. Biofilm formation was measured and antibiotic resistance was compared between before and after of biofilm formation. The polymerase chain reaction (PCR) and Real-Time PCR were performed to detect of abaI and pgaD genes. The biofilm producer isolates and the most resistant isolates were exposed to ozone gas .More than 70% strains were resistance to Erythromycin, Ofloxacin, Ceftazidime, Ceftriaxone, and Ticarcillin-clavulanic acid and 50% isolates were resistant to Imipenem. Thirty one (48.4%) isolates were biofilm producer. The pgaD and abaI genes were positive in 29 (45.3%) and 9 (14%) isolates, respectively. Real time PCR demonstrated that the copy numbers of the pgaD and abaI genes after biofilm formation were increased. After exposure to ozone, biofilm formation reduced in all very strong biofilm producing isolates. Our results showed that after biofilm formation, an increased resistance was observed in most isolates. Also rising expression of abaI gene was associated with biofilm formation and an increase of antibiotic resistance. In the current study, both biofilm formation and antibiotic resistance were reduced after O3 exposure.
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- Golberg A, Broelsch GF, Vecchio D, Khan S, Hamblin MR, Austen WG, Jr., et al. Eradication of multidrug-resistant A. baumannii in burn wounds by antiseptic pulsed electric field. TECHN. 2014;2(2):153-60.
- Perez F, Hujer AM, Hujer KM, Decker BK, Rather PN, Bonomo RA. Global challenge of multidrug-resistant Acinetobacter baumannii. AAC. 2007;51(10):3471-84.
- Wieczorek P, Sacha P, Hauschild T, Zorawski M, Krawczyk M, Tryniszewska E. Multidrug resistant Acinetobacter baumannii--the role of AdeABC (RND family) efflux pump in resistance to antibiotics. FOLIA HISTOCHEM CYTOBIOL. 2008;46(3):257-67.
- Yoon EJ, Chabane YN, Goussard S, Snesrud E, Courvalin P, De E, et al. Contribution of resistance-nodulation-cell division efflux systems to antibiotic resistance and biofilm formation in Acinetobacter baumannii. mBio. 2015;6(2).
- Stoodley P, Sauer K, Davies DG, Costerton JW. Biofilms as complex differentiated communities. ANNU REV. 2002;56:187-209.
- Zhang Y, Zhu Y, Gupta A, Huang Y, Murray CK, Vrahas MS, et al. Antimicrobial blue light therapy for multidrug-resistant Acinetobacter baumannii infection in a mouse burn model: implications for prophylaxis and treatment of combat-related wound infections. J INT J INFECT DIS. 2014;209(12):1963-71.
- Fontes B, Cattani Heimbecker AM, de Souza Brito G, Costa SF, van der Heijden IM, Levin AS, et al. Effect of low-dose gaseous ozone on pathogenic bacteria. BMC INFECT DIS. 2012;12:358.
- Fletcher LA, Gaunt LF, Beggs CB, Shepherd SJ, Sleigh PA, Noakes CJ, et al. Bactericidal action of positive and negative ions in air. BMC MICROBIOL. 2007;7:32.
- Fan L, Song J, Hildebrand PD, Forney CF. Interaction of ozone and negative air ions to control micro-organisms. J APPL MICROBIOL. 2002;93(1):144-8.
- Sharma M, Hudson JB. Ozone gas is an effective and practical antibacterial agent. AMER J INFECT CONTROL. 2008;36(8):559-63.
- (CLSI) CaLSI. Performance standards for antimicrobial susceptibility testing. editor 2011:M100-S20.
- O'Toole GA. Microtiter dish biofilm formation assay. JoVE. 2011(47).
- Hatami R. The frequency of multidrug-resistance and extensively drug-resistant Acinetobacter baumannii in west of Iran. J Clin Microbiol Infect Dis 2018;1(1):1-8.
- Askarian M, Hosseini RS, Kheirandish P, Assadian O. Incidence and outcome of nosocomial infections in female burn patients in Shiraz, Iran. AMER J INFECT CONTROL. 2004;32(1):23-6.
- Lee NY, Lee HC, Ko NY, Chang CM, Shih HI, Wu CJ, et al. Clinical and economic impact of multidrug resistance in nosocomial Acinetobacter baumannii bacteremia. INFECT CONTROL HOSP EPIDEMIOL. 2007;28(6):713-9.
- Taherikalani M, Maleki A, Sadeghifard N, Mohammadzadeh D, Soroush S, Asadollahi P, et al. Dissemination of class 1, 2 and 3 integrons among different multidrug resistant isolates of Acinetobacter baumannii in Tehran hospitals, Iran. POL J MICROBIOL. 2011;60(2):169-74.
- Rastegar-Lari A, Mohammadi-Barzelighi H, Arjomandzadegan M, Nosrati R, Owlia P. Distribution of Class I Integron among Isolates of Acinetobacter baumannii Recoverd from Burn Patients. J MED BACTERIOL. 2013;2(1-2):1-11.
- Oh JY, Kim KS, Jeong YW, Cho JW, Park JC, Lee JC. Epidemiological typing and prevalence of integrons in multiresistant Acinetobacter strains. APMIS. 2002;110:247-52.
- Moammad F, Arabestani MR, Safari M, Roshanaii G, Alikhani MY. Prevalence of class1, 2 and 3 integrons among extensive drug resistance Acinetobacter baumanii strains isolated from intensive care units in Hamadan, west province, Iran. J Med Microbiol. 2014;8(3):8-14.
- Kamal beik S, Kouchek M, Baseri Salehi M, Fallah F, Malekan MA, Talaie H. revalence of Class 2 Integrons in Multidrug-Resistant Acinetobacter Baumannii in Toxicological ICU Patients in Tehran. IRAN J TOXICOL. 2013;7(2):900-6.
- Abdi-Ali A, Hendiani S, Mohammadi P, Gharavi S. Assessment of Biofilm Formation and Resistance to Imipenem and Ciprofloxacin among Clinical Isolates of Acinetobacter baumannii in Tehran. JUNDISHAPUR J. 2014;7(1):e8606.
- Bardbari AM, Arabestani MR, Karami M, Keramat F, Alikhani MY, Bagheri KP. Correlation between ability of biofilm formation with their responsible genes and MDR patterns in clinical and environmental Acinetobacter baumannii isolates. MICROB. 2017;108:122-8.
- Rao RS, Karthika RU, Singh SP, Shashikala P, Kanungo R, Jayachandran S, et al. Correlation between biofilm production and multiple drug resistance in imipenem resistant clinical isolates of Acinetobacter baumannii. INDIAN J MED. 2008;26(4):333-7.
- Xiang J, Sun Z, Song F, Huan JN. [Expressions of pgaABC gene clusters and changes in biofilm phenotype of Acinetobacter baumannii in burn patients]. Zhonghua shao shang za zhi = Zhonghua shaoshang zazhi = CHINESE J BURN. 2011;27(2):100-3.
- Tong YQ, Dou L, Sun Y, Zhao D. Herb-resistant upec strains have different biofilm formation abilities and different expression of certain biofilm related genes. Afr J Tradit Complement Altern Med. 2017;14(5):1-9.
- Xiang J, Sun Z, Yang XG, Huan JN. [Changes in expression of gene aba I in biofilm of Acinetobacter baumannii strains isolated from burn patients]. Zhonghua shao shang za zhi = Zhonghua shaoshang zazhi = CHINESE J BURN. 2012;28(2):101-5.
- Anbazhagan D, Mansor M, Yan GO, Md Yusof MY, Hassan H, Sekaran SD. Detection of quorum sensing signal molecules and identification of an autoinducer synthase gene among biofilm forming clinical isolates of Acinetobacter spp. PloS one. 2012;7(7):e36696.
- Peleg AY, Jara S, Monga D, Eliopoulos GM, Moellering RC, Jr., Mylonakis E. Galleria mellonella as a model system to study Acinetobacter baumannii pathogenesis and therapeutics. AAC. 2009;53(6):2605-9.
- Kowalski WJ, Bahnfleth WP, Whittam TS. Bactericidal effects of high airborne ozone concentrations on escherichia coli and staphylococcus aureus. Ozone Sci Eng. 1998;20:205-21.
- Summerfelt ST. Ozonation and UV irradiation - an introduction and examples of current applications. Aquac Eng. 2003(28):21-36.
- Rickloff JR. An evaluation of the sporicidal activity of ozone. Appl Environ Microbiol 1987;53:683-6.
- Facile N, Barbeau B, Prevost M, Koudjonou B. Evaluating bacterial aerobic spores as a surrogate for giardia and cryptosporidium inactivation by ozone. Water Res. 2000;34(12):3238-46.
- Larson MA, Marinas BJ. Inactivation of Bacillus subtilis spores with ozone and monochloramine. Water Res. 2003;37:833-44.
- Martinez-Sanchez G, Al-Dalain SM, Menendez S, Re L, Giuliani A, Candelario-Jalil E, et al. Therapeutic efficacy of ozone in patients with diabetic foot. EUR J PHARMACOL. 2005;523(1-3):151-61.
- Bialoszewski D, Kowalewski M. Superficially, longer, intermittent ozone theraphy in the treatment of the chronic, infected wounds. Ortop Traumatol Rehabil. 2003;5(5):652-8.
References
Golberg A, Broelsch GF, Vecchio D, Khan S, Hamblin MR, Austen WG, Jr., et al. Eradication of multidrug-resistant A. baumannii in burn wounds by antiseptic pulsed electric field. TECHN. 2014;2(2):153-60.
Perez F, Hujer AM, Hujer KM, Decker BK, Rather PN, Bonomo RA. Global challenge of multidrug-resistant Acinetobacter baumannii. AAC. 2007;51(10):3471-84.
Wieczorek P, Sacha P, Hauschild T, Zorawski M, Krawczyk M, Tryniszewska E. Multidrug resistant Acinetobacter baumannii--the role of AdeABC (RND family) efflux pump in resistance to antibiotics. FOLIA HISTOCHEM CYTOBIOL. 2008;46(3):257-67.
Yoon EJ, Chabane YN, Goussard S, Snesrud E, Courvalin P, De E, et al. Contribution of resistance-nodulation-cell division efflux systems to antibiotic resistance and biofilm formation in Acinetobacter baumannii. mBio. 2015;6(2).
Stoodley P, Sauer K, Davies DG, Costerton JW. Biofilms as complex differentiated communities. ANNU REV. 2002;56:187-209.
Zhang Y, Zhu Y, Gupta A, Huang Y, Murray CK, Vrahas MS, et al. Antimicrobial blue light therapy for multidrug-resistant Acinetobacter baumannii infection in a mouse burn model: implications for prophylaxis and treatment of combat-related wound infections. J INT J INFECT DIS. 2014;209(12):1963-71.
Fontes B, Cattani Heimbecker AM, de Souza Brito G, Costa SF, van der Heijden IM, Levin AS, et al. Effect of low-dose gaseous ozone on pathogenic bacteria. BMC INFECT DIS. 2012;12:358.
Fletcher LA, Gaunt LF, Beggs CB, Shepherd SJ, Sleigh PA, Noakes CJ, et al. Bactericidal action of positive and negative ions in air. BMC MICROBIOL. 2007;7:32.
Fan L, Song J, Hildebrand PD, Forney CF. Interaction of ozone and negative air ions to control micro-organisms. J APPL MICROBIOL. 2002;93(1):144-8.
Sharma M, Hudson JB. Ozone gas is an effective and practical antibacterial agent. AMER J INFECT CONTROL. 2008;36(8):559-63.
(CLSI) CaLSI. Performance standards for antimicrobial susceptibility testing. editor 2011:M100-S20.
O'Toole GA. Microtiter dish biofilm formation assay. JoVE. 2011(47).
Hatami R. The frequency of multidrug-resistance and extensively drug-resistant Acinetobacter baumannii in west of Iran. J Clin Microbiol Infect Dis 2018;1(1):1-8.
Askarian M, Hosseini RS, Kheirandish P, Assadian O. Incidence and outcome of nosocomial infections in female burn patients in Shiraz, Iran. AMER J INFECT CONTROL. 2004;32(1):23-6.
Lee NY, Lee HC, Ko NY, Chang CM, Shih HI, Wu CJ, et al. Clinical and economic impact of multidrug resistance in nosocomial Acinetobacter baumannii bacteremia. INFECT CONTROL HOSP EPIDEMIOL. 2007;28(6):713-9.
Taherikalani M, Maleki A, Sadeghifard N, Mohammadzadeh D, Soroush S, Asadollahi P, et al. Dissemination of class 1, 2 and 3 integrons among different multidrug resistant isolates of Acinetobacter baumannii in Tehran hospitals, Iran. POL J MICROBIOL. 2011;60(2):169-74.
Rastegar-Lari A, Mohammadi-Barzelighi H, Arjomandzadegan M, Nosrati R, Owlia P. Distribution of Class I Integron among Isolates of Acinetobacter baumannii Recoverd from Burn Patients. J MED BACTERIOL. 2013;2(1-2):1-11.
Oh JY, Kim KS, Jeong YW, Cho JW, Park JC, Lee JC. Epidemiological typing and prevalence of integrons in multiresistant Acinetobacter strains. APMIS. 2002;110:247-52.
Moammad F, Arabestani MR, Safari M, Roshanaii G, Alikhani MY. Prevalence of class1, 2 and 3 integrons among extensive drug resistance Acinetobacter baumanii strains isolated from intensive care units in Hamadan, west province, Iran. J Med Microbiol. 2014;8(3):8-14.
Kamal beik S, Kouchek M, Baseri Salehi M, Fallah F, Malekan MA, Talaie H. revalence of Class 2 Integrons in Multidrug-Resistant Acinetobacter Baumannii in Toxicological ICU Patients in Tehran. IRAN J TOXICOL. 2013;7(2):900-6.
Abdi-Ali A, Hendiani S, Mohammadi P, Gharavi S. Assessment of Biofilm Formation and Resistance to Imipenem and Ciprofloxacin among Clinical Isolates of Acinetobacter baumannii in Tehran. JUNDISHAPUR J. 2014;7(1):e8606.
Bardbari AM, Arabestani MR, Karami M, Keramat F, Alikhani MY, Bagheri KP. Correlation between ability of biofilm formation with their responsible genes and MDR patterns in clinical and environmental Acinetobacter baumannii isolates. MICROB. 2017;108:122-8.
Rao RS, Karthika RU, Singh SP, Shashikala P, Kanungo R, Jayachandran S, et al. Correlation between biofilm production and multiple drug resistance in imipenem resistant clinical isolates of Acinetobacter baumannii. INDIAN J MED. 2008;26(4):333-7.
Xiang J, Sun Z, Song F, Huan JN. [Expressions of pgaABC gene clusters and changes in biofilm phenotype of Acinetobacter baumannii in burn patients]. Zhonghua shao shang za zhi = Zhonghua shaoshang zazhi = CHINESE J BURN. 2011;27(2):100-3.
Tong YQ, Dou L, Sun Y, Zhao D. Herb-resistant upec strains have different biofilm formation abilities and different expression of certain biofilm related genes. Afr J Tradit Complement Altern Med. 2017;14(5):1-9.
Xiang J, Sun Z, Yang XG, Huan JN. [Changes in expression of gene aba I in biofilm of Acinetobacter baumannii strains isolated from burn patients]. Zhonghua shao shang za zhi = Zhonghua shaoshang zazhi = CHINESE J BURN. 2012;28(2):101-5.
Anbazhagan D, Mansor M, Yan GO, Md Yusof MY, Hassan H, Sekaran SD. Detection of quorum sensing signal molecules and identification of an autoinducer synthase gene among biofilm forming clinical isolates of Acinetobacter spp. PloS one. 2012;7(7):e36696.
Peleg AY, Jara S, Monga D, Eliopoulos GM, Moellering RC, Jr., Mylonakis E. Galleria mellonella as a model system to study Acinetobacter baumannii pathogenesis and therapeutics. AAC. 2009;53(6):2605-9.
Kowalski WJ, Bahnfleth WP, Whittam TS. Bactericidal effects of high airborne ozone concentrations on escherichia coli and staphylococcus aureus. Ozone Sci Eng. 1998;20:205-21.
Summerfelt ST. Ozonation and UV irradiation - an introduction and examples of current applications. Aquac Eng. 2003(28):21-36.
Rickloff JR. An evaluation of the sporicidal activity of ozone. Appl Environ Microbiol 1987;53:683-6.
Facile N, Barbeau B, Prevost M, Koudjonou B. Evaluating bacterial aerobic spores as a surrogate for giardia and cryptosporidium inactivation by ozone. Water Res. 2000;34(12):3238-46.
Larson MA, Marinas BJ. Inactivation of Bacillus subtilis spores with ozone and monochloramine. Water Res. 2003;37:833-44.
Martinez-Sanchez G, Al-Dalain SM, Menendez S, Re L, Giuliani A, Candelario-Jalil E, et al. Therapeutic efficacy of ozone in patients with diabetic foot. EUR J PHARMACOL. 2005;523(1-3):151-61.
Bialoszewski D, Kowalewski M. Superficially, longer, intermittent ozone theraphy in the treatment of the chronic, infected wounds. Ortop Traumatol Rehabil. 2003;5(5):652-8.