Detection of antibiotics resistance andefflux pumps production in clinical Acinetobacter baumannii isolates
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Abstract
Acinetobacter baumannii is considered one of the important pathogenic bacteria and responsible for several nosocomial and community infections in humans, such as the lungs (pneumonia), the bloodstream of ICU patients, burns, surgical wounds and meningitis. During the period from September to the end of December 2022, a total of 210 blood specimens were collected inDiyala governorate Baqubah Teaching Hospital, Iraq.To investigate the prevalence of bacterial isolates among ICU patients by identifying bacteria depending on suitable media under suitable environmental conditions for growth, morphological characteristics, biochemical tests, and confirmation of identification isolates by molecular detection of blaOXA- 51gene, antibiotic resistance, phenotypic and molecular detection of efflux pumps adeF gene, and activity of ciprofloxacin andresveratrol on gene expression. The percentage positive growth of A. baumannii isolates from blood specimens among ICU patients was 12 %, and the highest resistance of 25 isolates against fourteen types of antibiotics was equal to cefotaxime (100 % ), ticarcillin-clavulanate, and ceftriaxone ( 96 %), and ( 92 % )for each of Cefepime and Tobramycin, the lowest percentage was for doxycycline (64 % ).MIC value for ciprofloxacin and doxycycline between (8- 512 mg/ml) and ( 4 - 128 mg/ml) respectively. Molecular studies indicate the presence of blaOXA- 51gene and adeF in all isolates under study ( 100 % ). The treatment of resistance isolates with ciprofloxacin, resveratrol and their combination resulted in reduction of adeF gene. The resveratrol effect on efflux pumps gene adeF gene expression occurred for the first time in Iraq and even at the level of the Arab world.
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Keywords
Acinetobacter baumannii, AdeF gene, Antibiotic, Efflux pump inhibitors
References
AL-Dahlaki, S. M. M. (2020). Molecular detection and gene expression for hcp and blaOXA-51 genes in Acinetobacter baumannii isolated from different clinical sources (Doctoral dissertation, MSC Thesis, College of Science, University of Diyala, Diyala, Iraq, 1-154.
Al-Shamiri, M. M., Zhang, S., Mi, P., Liu, Y., Xun, M., Yang, E., ... & Chen, Y. (2021). Phenotypic and genotypic characteristics of Acinetobacter baumannii enrolled in the relationship among antibiotic resistance, biofilm formation and motility, Microbial Pathogenesis, 155(4), 104922-104932. https://doi.org/10.1016/j.micpath.2021.104922.
AL Hadeedy, U. E. I., Mahdi, N. B. & AL Jebory, I. S. (2019). Isolation and Diagnosis of Acinetobacter Bumanniii Recently Isolated From Patients in Kirkuk Hospitals and Study their Antibiotics Resistance. Kirkuk University Journal-Scientific Studies, 14(3), 155-173. https://doi.org/10.32894/kujss.2019.14.3.12.
Ali, M. A. & SS Mahmood. 2022. 'Role of RND-efflux pump in levofloxacin resistance among Acinetobacter baumannii', Iraqi Journal of Agricultural Sciences, 53 (3),542-550.
Asif, M., Alvi, I. A. & Rehman, S. U. (2018). Insight into Acinetobacter baumannii: pathogenesis, global resistance, mechanisms of resistance, treatment options, and alternative modalities. Infection and drug resistance, 16 (9),1249-1260.
Bajpai, T., Bhatambare, G. S., Varma, M., & Pandey, M. (2016). Species-level identification of Acinetobacter by 16s rRNA sequencing: Necessity today, essentiality tomorrow. New Nigerian Journal of Clinical Research, 5(8), 55-65.http://doi.org/ 10.4103/2250-9658.197438i.
Cheesman, M. J., Ilanko, A., Blonk, B. & Cock, I. E. (2017). Developing new antimicrobial therapies: are synergistic combinations of plant extracts/compounds with conventional antibiotics the solution?.Pharmaco gnosyreviews, 11(22), 57-67.https://doi.org/10.4103%2Fphrev.ph rev_21_17
CLSI, (2022). Performance Standards for Antimicrobial Susceptibility Testing. 27th ed. Wayne, PA: Clinical and Laboratory Standards Institute.
Constantiniu, S., Romaniuc, A., Iancu, L. S., Filimon, R., & Taraşi, I. (2004). Cultural and biochemical characteristics of Acinetobacter spp. strains isolated from hospital units. The journal of preventive medicine, 12(34), 35-42.
Darweesh, A. O., Shareef, A. A. & Alriyahee, F. A. A. (2021). Isolation and Identification of Cellulytic Ruminant Pseudomonas Aeruginosa by Classical and Molecular Methods in Basrah Province. Annals of the Romanian Society for Cell Biology, 1(4),4672-4680.
Djankpa, F. T., Wiafe, G. A., Boateng, B. N., Tsegah, K. M., Essien-Baidoo, S., Ulanja, M. B., ... & Asante, G. (2021). Assessment of corn starch as substitute for agarose in DNA gel electrophoresis. BMC Research Notes, 14(1), 1-6https://doi.org/10.1186/s13104-021-05483-1.
Doi, Y., Murray, G. L. & Peleg, A. Y. (2015, February). Acinetobacter baumannii: evolution of antimicrobial resistance-treatment options. In Seminars in respiratory and critical care medicine, 1(36), 85-98. http://doi.org/ 10.1055/s-0034-1398388.
Fernando, D. M. (2015). Analysis of resistance nodulation division efflux pumps in Acinetobacter baumannii (Doctoral dissertation, University of Ontario Institute of Technology) 1- 133.
Havenga, B., Reyneke, B., Ndlovu, T. & Khan, W. (2022). Genotypic and phenotypic comparison of clinical and environmental Acinetobacter baumannii strains. Microbial Pathogenesis, 172(14), 1049-1057.https://doi.org/10.1016/j.micpath.2022.105749
Hood, M. I., Jacobs, A. C., Sayood, K., Dunman, P. M. & Skaar, E. P. (2010). Acinetobacter baumannii increases tolerance to antibiotics in response to monovalent cations. Antimicrobial agents and chemotherapy, 54(3), 1029-1041.https://doi.org/10.1128/aac.00963-09
Kadhom, H. & Ali, M. (2022). Epidemiological molecular analysis of Acinetobacter baumannii isolates using a multilocus sequencing typing and global lineage. Revis Bionatura, 7(1), 29.
Kadom, S. M. & ABID, I. N. (2020). Detection of bla OXA-51-like and bla VIM Carbapenemase Genes in Acinetobacter baumannii isolated from burn patients. International Journal of Pharmaceutical Research, 12(2), 23-35.
Kaviani, R., Pouladi, I., Niakan, M. & Mirnejad, R. (2020). Molecular detection of Adefg efflux pump genes and their contribution to antibiotic resistance in Acinetobacter baumannii clinical isolates. Reports of Biochemistry & Molecular Biology, 8(4), 413.
Kim, S. H., Yun, S., & Park, W. (2022). Constitutive Phenotypic Modification of Lipid A in Clinical Acinetobacter baumannii Isolates. Microbiology spectrum, 10(4), 1295-1322.https://doi.org/10.1128/spectrum.01295-22
Lee, H. Y., Hsu, S. Y., Hsu, J. F., Chen, C. L., Wang, Y. H. & Chiu, C. H. (2018). Risk factors and molecular epidemiology of Acinetobacter baumannii bacteremia in neonates. Journal of microbiology, immunology and infection, 51(3), 367-376. https://doi.org/10.1016/j.jmii.2017.07.013
Liu, L., Yu, J., Shen, X., Cao, X., Zhan, Q., Guo, Y.& Yu, F. (2020). Resveratrol enhances the antimicrobial effect of polymyxin B on Klebsiella pneumoniae and Escherichia coli isolates with polymyxin B resistance. BMC microbiology, 20 (1), 1-8.https://doi.org/10.1186/s12866-020-01995-1
Martins, M., Viveiros, M., Couto, I., Costa, S. S., Pacheco, T., Fanning, S., & Amaral, L. (2011). Identification of efflux pump-mediated multidrug-resistant bacteria by the ethidium bromide-agar cartwheel method. In vivo, 25(2), 171-178.
Monem, S., Furmanek-Blaszk, B., Łupkowska, A., Kuczyńska-Wiśnik, D., Stojowska-Swędrzyńska, K. & Laskowska, E. (2020). Mechanisms protecting Acinetobacter baumannii against multiple stresses triggered by the host immune response, antibiotics and outside-host environment. International Journal of Molecular Sciences, 21(15), 5498-5510. https://doi.org/10.3390/ijms21155498
Muhammad, M. J., Mubarak, K. I. & Wadi, K. D. (2023). Study of Antibacterial Activity from Alcoholic Extracts of Torilis arvensis (Huds.) Link on Biofilm formation of Acinetobacter baumannii in Diyala Province-Iraq. Academic Science Journal, 1(1), 60-76.https://doi.org/10.24237/ASJ.01.01.614C
Nasr, P. (2020). Genetics, epidemiology, and clinical manifestations of multidrug-resistant Acinetobacter baumannii. Journal of Hospital Infection, 104(1), 4-11. https://doi.org/10.1016/j.jhin.2019.09.021
PEYMAnI, A. M. I. R., FARAJnIA, S. A. F. A. R., Nahaei, M. R., Sohrabi, N., Abbasi, L. A. L. E. H., Ansarin, K. H. A. L. I. L. & Azhari, F. A. T. E. M. E. H. (2012). Prevalence of class 1 integron among multidrug-resistant Acinetobacter baumannii in Tabriz, northwest of Iran. Polish Journal of Microbiology, 61(1), 57-60.
Ridha, D. J., Ali, M. R. & Jassim, K. A. (2019). Molecular analysis of multi–drug resistant Acinetobacter baumannii isolated in Baghdad hospital. Int. J. Res. Pharm. Sci, 10(3), 1970-1987.
Rovelli, S. (1998). The role of polysaccharides in plasmodium falciparum malaria (Doctoral dissertation), 1-185.
Salimizand, H., Zomorodi, A. R., Mansury, D., Khakshoor, M., Azizi, O., Khodaparast, S., & Sadeghian, H. (2018). Diversity of aminoglycoside modifying enzymes and 16S rRNA methylases in Acinetobacter baumannii and Acinetobacter nosocomialis species in Iran; wide distribution of aadA1 and armA. Infection, Genetics and Evolution, 66 (1), 195-199.https://doi.org/10.1016/j.meegid.2018.09.028
Shuster, Y., Steiner-Mordoch, S., Alon Cudkowicz, N., & Schuldiner, S. (2016). A transporter interactome is essential for the cquisition of antimicrobial resistance to antibiotics. PLoS One, 11(4),55-66.https://doi.org/10.1371/journal.pone.0152917
Subramanian Swaminathan, G. I., Arora, S., Mehta, S., Mittal, R., Mane, A. & Charugulla, S. N. (2020). Susceptibility pattern of doxycycline in comparison to azithromycin, cefuroxime and amoxicillin against common isolates: a retrospective study based on diagnostic laboratory data. Journal of The Association of Physicians of India, 68(1), P59.
Wang, Y., Su, J., Zhou, Z., Yang, J., Liu, W., Zhang, Y., ... & Li, G. (2023). Baicalein Resensitizes Multidrug-Resistant Gram-Negative Pathogens to Doxycycline. Microbiology Spectrum, 11(3), 4702-4722.https://doi.org/10.1128/spectrum.04702-22
Wareth, G., Brandt, C., Sprague, L. D., Neubauer, H. & Pletz, M. W. (2021). WGS based analysis of acquired antimicrobial resistance in human and non-human Acinetobacter baumannii isolates from a German perspective. BMC microbiology, 21(1), 1-13.https://doi.org/10.1186/s12866-021-02270-7
Xu, C., Bilya, S. R. & Xu, W. (2019). adeABC efflux gene in Acinetobacter baumannii. New Microbes New Infect, 30 (8), 100549.
Al-Shamiri, M. M., Zhang, S., Mi, P., Liu, Y., Xun, M., Yang, E., ... & Chen, Y. (2021). Phenotypic and genotypic characteristics of Acinetobacter baumannii enrolled in the relationship among antibiotic resistance, biofilm formation and motility, Microbial Pathogenesis, 155(4), 104922-104932. https://doi.org/10.1016/j.micpath.2021.104922.
AL Hadeedy, U. E. I., Mahdi, N. B. & AL Jebory, I. S. (2019). Isolation and Diagnosis of Acinetobacter Bumanniii Recently Isolated From Patients in Kirkuk Hospitals and Study their Antibiotics Resistance. Kirkuk University Journal-Scientific Studies, 14(3), 155-173. https://doi.org/10.32894/kujss.2019.14.3.12.
Ali, M. A. & SS Mahmood. 2022. 'Role of RND-efflux pump in levofloxacin resistance among Acinetobacter baumannii', Iraqi Journal of Agricultural Sciences, 53 (3),542-550.
Asif, M., Alvi, I. A. & Rehman, S. U. (2018). Insight into Acinetobacter baumannii: pathogenesis, global resistance, mechanisms of resistance, treatment options, and alternative modalities. Infection and drug resistance, 16 (9),1249-1260.
Bajpai, T., Bhatambare, G. S., Varma, M., & Pandey, M. (2016). Species-level identification of Acinetobacter by 16s rRNA sequencing: Necessity today, essentiality tomorrow. New Nigerian Journal of Clinical Research, 5(8), 55-65.http://doi.org/ 10.4103/2250-9658.197438i.
Cheesman, M. J., Ilanko, A., Blonk, B. & Cock, I. E. (2017). Developing new antimicrobial therapies: are synergistic combinations of plant extracts/compounds with conventional antibiotics the solution?.Pharmaco gnosyreviews, 11(22), 57-67.https://doi.org/10.4103%2Fphrev.ph rev_21_17
CLSI, (2022). Performance Standards for Antimicrobial Susceptibility Testing. 27th ed. Wayne, PA: Clinical and Laboratory Standards Institute.
Constantiniu, S., Romaniuc, A., Iancu, L. S., Filimon, R., & Taraşi, I. (2004). Cultural and biochemical characteristics of Acinetobacter spp. strains isolated from hospital units. The journal of preventive medicine, 12(34), 35-42.
Darweesh, A. O., Shareef, A. A. & Alriyahee, F. A. A. (2021). Isolation and Identification of Cellulytic Ruminant Pseudomonas Aeruginosa by Classical and Molecular Methods in Basrah Province. Annals of the Romanian Society for Cell Biology, 1(4),4672-4680.
Djankpa, F. T., Wiafe, G. A., Boateng, B. N., Tsegah, K. M., Essien-Baidoo, S., Ulanja, M. B., ... & Asante, G. (2021). Assessment of corn starch as substitute for agarose in DNA gel electrophoresis. BMC Research Notes, 14(1), 1-6https://doi.org/10.1186/s13104-021-05483-1.
Doi, Y., Murray, G. L. & Peleg, A. Y. (2015, February). Acinetobacter baumannii: evolution of antimicrobial resistance-treatment options. In Seminars in respiratory and critical care medicine, 1(36), 85-98. http://doi.org/ 10.1055/s-0034-1398388.
Fernando, D. M. (2015). Analysis of resistance nodulation division efflux pumps in Acinetobacter baumannii (Doctoral dissertation, University of Ontario Institute of Technology) 1- 133.
Havenga, B., Reyneke, B., Ndlovu, T. & Khan, W. (2022). Genotypic and phenotypic comparison of clinical and environmental Acinetobacter baumannii strains. Microbial Pathogenesis, 172(14), 1049-1057.https://doi.org/10.1016/j.micpath.2022.105749
Hood, M. I., Jacobs, A. C., Sayood, K., Dunman, P. M. & Skaar, E. P. (2010). Acinetobacter baumannii increases tolerance to antibiotics in response to monovalent cations. Antimicrobial agents and chemotherapy, 54(3), 1029-1041.https://doi.org/10.1128/aac.00963-09
Kadhom, H. & Ali, M. (2022). Epidemiological molecular analysis of Acinetobacter baumannii isolates using a multilocus sequencing typing and global lineage. Revis Bionatura, 7(1), 29.
Kadom, S. M. & ABID, I. N. (2020). Detection of bla OXA-51-like and bla VIM Carbapenemase Genes in Acinetobacter baumannii isolated from burn patients. International Journal of Pharmaceutical Research, 12(2), 23-35.
Kaviani, R., Pouladi, I., Niakan, M. & Mirnejad, R. (2020). Molecular detection of Adefg efflux pump genes and their contribution to antibiotic resistance in Acinetobacter baumannii clinical isolates. Reports of Biochemistry & Molecular Biology, 8(4), 413.
Kim, S. H., Yun, S., & Park, W. (2022). Constitutive Phenotypic Modification of Lipid A in Clinical Acinetobacter baumannii Isolates. Microbiology spectrum, 10(4), 1295-1322.https://doi.org/10.1128/spectrum.01295-22
Lee, H. Y., Hsu, S. Y., Hsu, J. F., Chen, C. L., Wang, Y. H. & Chiu, C. H. (2018). Risk factors and molecular epidemiology of Acinetobacter baumannii bacteremia in neonates. Journal of microbiology, immunology and infection, 51(3), 367-376. https://doi.org/10.1016/j.jmii.2017.07.013
Liu, L., Yu, J., Shen, X., Cao, X., Zhan, Q., Guo, Y.& Yu, F. (2020). Resveratrol enhances the antimicrobial effect of polymyxin B on Klebsiella pneumoniae and Escherichia coli isolates with polymyxin B resistance. BMC microbiology, 20 (1), 1-8.https://doi.org/10.1186/s12866-020-01995-1
Martins, M., Viveiros, M., Couto, I., Costa, S. S., Pacheco, T., Fanning, S., & Amaral, L. (2011). Identification of efflux pump-mediated multidrug-resistant bacteria by the ethidium bromide-agar cartwheel method. In vivo, 25(2), 171-178.
Monem, S., Furmanek-Blaszk, B., Łupkowska, A., Kuczyńska-Wiśnik, D., Stojowska-Swędrzyńska, K. & Laskowska, E. (2020). Mechanisms protecting Acinetobacter baumannii against multiple stresses triggered by the host immune response, antibiotics and outside-host environment. International Journal of Molecular Sciences, 21(15), 5498-5510. https://doi.org/10.3390/ijms21155498
Muhammad, M. J., Mubarak, K. I. & Wadi, K. D. (2023). Study of Antibacterial Activity from Alcoholic Extracts of Torilis arvensis (Huds.) Link on Biofilm formation of Acinetobacter baumannii in Diyala Province-Iraq. Academic Science Journal, 1(1), 60-76.https://doi.org/10.24237/ASJ.01.01.614C
Nasr, P. (2020). Genetics, epidemiology, and clinical manifestations of multidrug-resistant Acinetobacter baumannii. Journal of Hospital Infection, 104(1), 4-11. https://doi.org/10.1016/j.jhin.2019.09.021
PEYMAnI, A. M. I. R., FARAJnIA, S. A. F. A. R., Nahaei, M. R., Sohrabi, N., Abbasi, L. A. L. E. H., Ansarin, K. H. A. L. I. L. & Azhari, F. A. T. E. M. E. H. (2012). Prevalence of class 1 integron among multidrug-resistant Acinetobacter baumannii in Tabriz, northwest of Iran. Polish Journal of Microbiology, 61(1), 57-60.
Ridha, D. J., Ali, M. R. & Jassim, K. A. (2019). Molecular analysis of multi–drug resistant Acinetobacter baumannii isolated in Baghdad hospital. Int. J. Res. Pharm. Sci, 10(3), 1970-1987.
Rovelli, S. (1998). The role of polysaccharides in plasmodium falciparum malaria (Doctoral dissertation), 1-185.
Salimizand, H., Zomorodi, A. R., Mansury, D., Khakshoor, M., Azizi, O., Khodaparast, S., & Sadeghian, H. (2018). Diversity of aminoglycoside modifying enzymes and 16S rRNA methylases in Acinetobacter baumannii and Acinetobacter nosocomialis species in Iran; wide distribution of aadA1 and armA. Infection, Genetics and Evolution, 66 (1), 195-199.https://doi.org/10.1016/j.meegid.2018.09.028
Shuster, Y., Steiner-Mordoch, S., Alon Cudkowicz, N., & Schuldiner, S. (2016). A transporter interactome is essential for the cquisition of antimicrobial resistance to antibiotics. PLoS One, 11(4),55-66.https://doi.org/10.1371/journal.pone.0152917
Subramanian Swaminathan, G. I., Arora, S., Mehta, S., Mittal, R., Mane, A. & Charugulla, S. N. (2020). Susceptibility pattern of doxycycline in comparison to azithromycin, cefuroxime and amoxicillin against common isolates: a retrospective study based on diagnostic laboratory data. Journal of The Association of Physicians of India, 68(1), P59.
Wang, Y., Su, J., Zhou, Z., Yang, J., Liu, W., Zhang, Y., ... & Li, G. (2023). Baicalein Resensitizes Multidrug-Resistant Gram-Negative Pathogens to Doxycycline. Microbiology Spectrum, 11(3), 4702-4722.https://doi.org/10.1128/spectrum.04702-22
Wareth, G., Brandt, C., Sprague, L. D., Neubauer, H. & Pletz, M. W. (2021). WGS based analysis of acquired antimicrobial resistance in human and non-human Acinetobacter baumannii isolates from a German perspective. BMC microbiology, 21(1), 1-13.https://doi.org/10.1186/s12866-021-02270-7
Xu, C., Bilya, S. R. & Xu, W. (2019). adeABC efflux gene in Acinetobacter baumannii. New Microbes New Infect, 30 (8), 100549.
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Detection of antibiotics resistance andefflux pumps production in clinical Acinetobacter baumannii isolates. (2023). Journal of Applied and Natural Science, 15(4), 1347-1353. https://doi.org/10.31018/jans.v15i4.4965
