Research Article
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Antibiotic susceptibility and biofilm formation of multi-drug resistant Gram-negative bacteria

Year 2023, , 45 - 50, 28.04.2023
https://doi.org/10.26650/IstanbulJPharm.2023.1156339

Abstract

Background and Aims: Gram-negative bacteria are important pathogens that can cause community- and hospital-acquired infections as well as opportunistic infections, with antimicrobial resistance in Gram-negative bacteria becoming a growing crisis in clinical medicine. Biofilm formation is one of the mechanisms of bacterial resistance, which makes bacteria less susceptible to antimicrobial agents and unable to be killed by host immune mechanisms. Therefore, this study investigates the antimicrobial resistances and biofilm-forming abilities of a total of 98 Gram-negative strains isolated from various clinical specimens.
Methods: A disc diffusion assay was performed to detect the susceptibility profiles of 98 Gram-negative strains. The biofilm- forming abilities of strains were also determined using the Crystal Violet assay.
Results: Concerning the disc diffusion assay, most of the isolates were found to be resistant to carbapenems, with more than 90% of Acinetobacter baumannii, Klebsiella pneumoniae and Escherichia coli isolates being found resistant to ceftazidime and piperacillin. Most of the Pseudomonas aeruginosa isolates (75%) were found to be resistant to imipenem and aztreonam. All isolates had the ability to form biofilms. Overall, 56% of isolates were strong formers, and 29% were moderate biofilm formers. Strong biofilm formation was observed in most strains except for K. pneumoniae.
Conclusion: The surveillance of susceptibility profiles and biofilm formation is important for determining their variable sus- ceptibility patterns and aiding in the appropriate management of infections caused by these organisms.

Supporting Institution

Research Fund of Istanbul University

Project Number

37825

References

  • Avcıoğlu, F. & Behçet, M. (2020). Üriner sistem enfeksiyonu et- keni Escherichia coli izolatlarının çeşitli antibiyotiklere direnç oranlarının değerlendirilmesi. Türk Mikrobiyoloji Cemiyeti Dergisi, 50(3), 172-7.
  • Bader, M. S., Loeb, M., Leto, D., & Brooks, A. A. (2020). Treatment of urinary tract infections in the era of antimicrobial resistance and new antimicrobial agents. Postgraduate Medicine, 132(3), 234–250. https://doi.org/10.1080/00325481.2019.1680052
  • Breijyeh, Z., Jubeh, B., & Karaman, R. (2020). Resistance of gram- negative bacteria to current antibacterial agents and approaches to resolve it. Molecules, 25(6), 1340.
  • Candan, E. D., & Aksöz, N. (2015). Klebsiella pneumoniae: charac- teristics of carbapenem resistance and virulence factors. Acta Biochimica Polonica, 62(4), 867–874. https://doi.org/10.18388/ abp.2015_1148
  • Center for Disease Control and Prevention (CDC). Antibiotic Resis- tance Threats in the United States, 2019. Atlanta, GA: U.S. Depart- ment of Health and Human Services, CDC; 2019.
  • Cepas, V., López, Y., Muñoz, E., Rolo, D., Ardanuy, C., Martí, S., Xer- cavins, M., Horcajada, J. P., Bosch, J., & Soto, S. M. (2019). Relation- ship between biofilm formation and antimicrobial resistance in Gram-negative bacteria. Microbial Drug Resistance (Larchmont, N.Y.), 25(1), 72–79. https://doi.org/10.1089/mdr.2018.0027
  • Chung, H.S., Hong, SG., Lee, Y., Kim, M., Yong, D. Jeong, SH., Lee, K. & Chong, Y. (2012). Antimicrobial susceptibility of Stenotrophomon- as maltophilia isolates from a Korean tertiary care hospital. Yon- sei Medical Journal 53(2), 439-41. doi: 10.3349/ymj.2012.53.2.439. PMID: 22318836; PMCID: PMC3282950.
  • Clinical and Laboratory Standards Institute (CLSI) (2021) Perfor- mance Standards for Antimicrobial. Document M100-Ed.31., Wayne, PA: CLSI.
  • Del Pozo J. L. (2018). Biofilm-related disease. Expert Review of Anti- Infective Therapy, 16(1), 51–65. https://doi.org/10.1080/14787210.2018.1417036
  • Dosler, S. & Karaaslan E. (2014). Inhibition and destruction of Pseu- domonas aeruginosa biofilms by antibiotics and antimicrobial peptides. Peptides, 62, 32-37.
  • Dumaru, R., Baral, R., & Shrestha, L. B. (2019). Study of biofilm for- mation and antibiotic resistance pattern of gram-negative Ba- cilli among the clinical isolates at BPKIHS, Dharan. BMC Research Notes, 12(1), 38. https://doi.org/10.1186/s13104-019-4084-8
  • Eichenberger, E. M., & Thaden, J. T. (2019). Epidemiology and mechanisms of resistance of extensively drug resistant Gram- negative bacteria. Antibiotics, 8(2), 37.
  • El-Gamal, M. I., Brahim, I., Hisham, N., Aladdin, R., Mohammed, H., & Bahaaeldin, A. (2017). Recent updates of carbapenem antibiot- ics. European Journal of Medicinal Chemistry, 131, 185–195. https:// doi.org/10.1016/j.ejmech.2017.03.022
  • Hammoudi Halat, D., & Ayoub Moubareck, C. (2020). The current burden of carbapenemases: Review of significant properties and dissemination among Gram-negative bacteria. Antibiotics, 9(4), 186.
  • İnce, G., Mirza, H. C., Üsküdar Güçlü, A., Gümüş, H., Erol, Ç., & Başustaoğlu, A. (2021). Comparison of in vitro activities of plazo- micin and other aminoglycosides against clinical isolates of Kleb- siella pneumoniae and Escherichia coli. The Journal of Antimicrobial Chemotherapy, 76(12), 3192–3196. https://doi.org/10.1093/jac/ dkab331
  • Jamal, M., Ahmad, W., Andleeb, S., Jalil, F., Imran, M., Nawaz, M. A., Hussain, T., Ali, M., Rafiq, M., & Kamil, M. A. (2018). Bacterial biofilm and associated infections. Journal of the Chinese Medical Associa- tion: JCMA, 81(1), 7–11. https://doi.org/10.1016/j.jcma.2017.07.012
  • Jia, X., Ma, W., He, J., Tian, X., Liu, H., Zou, H., & Cheng, S. (2020).
  • Heteroresistance to cefepime in Pseudomonas aeruginosa bacter- aemia. International Journal of Antimicrobial Agents, 55(3), 105832. https://doi.org/10.1016/j.ijantimicag.2019.10.013
  • Magiorakos, A. P., Srinivasan, A., Carey, R. B., Carmeli, Y., Falagas, M. E., Giske, C. G., & Monnet, D. L. (2012). Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clinical Microbiology and Infection, 18(3), 268- 281.
  • Malone, L., & Kwon, D. H. (2013). Carbapenem-associated mul- tidrug-resistant Acinetobacter baumannii are sensitised by az- treonam in combination with polyamines. International journal of Antimicrobial Agents, 41(1), 70–74. https://doi.org/10.1016/j. ijantimicag.2012.08.009
  • Mirza, H. C., Sancak, B., (2020). Comparison of antimicrobial sus- ceptibilities of Escherichia coli isolated from urinary cultures of different patient groups: A university hospital experience. Klimik Dergisi, 33(3), 270-6.
  • Mojica, M. F., Humphries, R., Lipuma, J. J., Mathers, A. J., Rao, G. G., Shelburne, S. A., Fouts, D. E., Van Duin, D., & Bonomo, R. A. (2022). Clinical challenges treating Stenotrophomonas maltophilia in- fections: an update. JAC-Antimicrobial Resistance, 4(3), dlac040. https://doi.org/10.1093/jacamr/dlac040
  • Namaei, M. H., Yousefi, M., Askari, P., Roshanravan, B., Hashemi, A., & Rezaei, Y. (2021). High prevalence of multidrug-resistant non-fermentative Gram-negative bacilli harboring blaIMP-1 and blaVIM-1 metallo-beta-lactamase genes in Birjand, south-east Iran. Iranian Journal of Microbiology, 13(4), 470–479. https://doi. org/10.18502/ijm.v13i4.6971
  • Nirwati, H., Sinanjung, K., Fahrunissa, F., Wijaya, F., Napitupulu, S., Hati, V. P., Hakim, M. S., Meliala, A., Aman A. T., & Nuryastuti, T. (2019). Biofilm formation and antibiotic resistance of Klebsiella pneumoniae isolated from clinical samples in a tertiary care hos- pital, Klaten, Indonesia. BMC Proceedings, 13(11), 1-8. https://doi. org/10.1186/s12919-019-0176-7
  • Pang, Z., Raudonis, R., Glick, B. R., Lin, T. J., & Cheng, Z. (2019). An- tibiotic resistance in Pseudomonas aeruginosa: mechanisms and alternative therapeutic strategies. Biotechnology Advances, 37(1), 177–192. https://doi.org/10.1016/j.biotechadv.2018.11.013
  • Peeters, E., Nelis, H. J. & Coenye. T., (2008). Comparison of mul- tiple methods for quantification of microbial biofilms grown in microtiter plates. Journal of Microbiology Methods, 72(2), 157-165. doi:10.1016/j.mimet.2007.11.010
  • Perdikouri, E. I. A., Arvaniti, K., Lathyris, D., Apostolidou Kiouti, F., Siskou, E., Haidich, A. B., & Papandreou, C. (2019). Infections due to multidrug-resistant bacteria in oncological patients: insights from a five-year epidemiological and clinical analysis. Microorgan- isms, 7(9), 277.
  • Peters, L., Olson, L., Khu, D. T., Linnros, S., Le, N. K., Hanberger, H. & Larsson, M. (2019). Multiple antibiotic resistance as a risk factor for mortality and prolonged hospital stay: a cohort study among neonatal intensive care patients with hospital-acquired infec- tions caused by Gram-negative bacteria in Vietnam. PloS one, 14(5), e0215666.
  • Rice, L. B. (2008). Federal funding for the study of antimicrobial resistance in nosocomial pathogens: no ESKAPE. The Journal of Infectious Diseases, 197(8), 1079-1081.
  • Thaden, J. T., Li, Y., Ruffin, F., Maskarinec, S. A., Hill-Rorie, J. M., Wan- da, L. C., & Fowler Jr, V. G. (2017). Increased costs associated with bloodstream infections caused by multidrug-resistant gram-neg- ative bacteria are due primarily to patients with hospital-acquired infections. Antimicrobial Agents and Chemotherapy, 61(3), e01709-16.
  • Vivas, R., Barbosa, A. A. T., Dolabela, S. S., & Jain, S. (2019). Multi- drug-resistant bacteria and alternative methods to control them: an overview. Microbial Drug Resistance, 25(6), 890-908.
  • Wang, G., Zhao, G., Chao, X., Xie, L., & Wang, H. (2020). The charac- teristic of virulence, biofilm and antibiotic resistance of Klebsiella pneumoniae. International Journal of Environmental Research and Public Health, 17(17), 6278.
  • World Health Organization (WHO) https://www.who.int/news- room/fact-sheets/detail/antimicrobial-resistance
  • Yadav, S.K., Bhujel, R. & Mishra, S.K. (2020). Emergence of multi- drug-resistant non-fermentative gram negative bacterial infec- tion in hospitalized patients in a tertiary care center of Nepal. BMC Res Notes, 13, 319. https://doi.org/10.1186/s13104-020-05163-6
Year 2023, , 45 - 50, 28.04.2023
https://doi.org/10.26650/IstanbulJPharm.2023.1156339

Abstract

Project Number

37825

References

  • Avcıoğlu, F. & Behçet, M. (2020). Üriner sistem enfeksiyonu et- keni Escherichia coli izolatlarının çeşitli antibiyotiklere direnç oranlarının değerlendirilmesi. Türk Mikrobiyoloji Cemiyeti Dergisi, 50(3), 172-7.
  • Bader, M. S., Loeb, M., Leto, D., & Brooks, A. A. (2020). Treatment of urinary tract infections in the era of antimicrobial resistance and new antimicrobial agents. Postgraduate Medicine, 132(3), 234–250. https://doi.org/10.1080/00325481.2019.1680052
  • Breijyeh, Z., Jubeh, B., & Karaman, R. (2020). Resistance of gram- negative bacteria to current antibacterial agents and approaches to resolve it. Molecules, 25(6), 1340.
  • Candan, E. D., & Aksöz, N. (2015). Klebsiella pneumoniae: charac- teristics of carbapenem resistance and virulence factors. Acta Biochimica Polonica, 62(4), 867–874. https://doi.org/10.18388/ abp.2015_1148
  • Center for Disease Control and Prevention (CDC). Antibiotic Resis- tance Threats in the United States, 2019. Atlanta, GA: U.S. Depart- ment of Health and Human Services, CDC; 2019.
  • Cepas, V., López, Y., Muñoz, E., Rolo, D., Ardanuy, C., Martí, S., Xer- cavins, M., Horcajada, J. P., Bosch, J., & Soto, S. M. (2019). Relation- ship between biofilm formation and antimicrobial resistance in Gram-negative bacteria. Microbial Drug Resistance (Larchmont, N.Y.), 25(1), 72–79. https://doi.org/10.1089/mdr.2018.0027
  • Chung, H.S., Hong, SG., Lee, Y., Kim, M., Yong, D. Jeong, SH., Lee, K. & Chong, Y. (2012). Antimicrobial susceptibility of Stenotrophomon- as maltophilia isolates from a Korean tertiary care hospital. Yon- sei Medical Journal 53(2), 439-41. doi: 10.3349/ymj.2012.53.2.439. PMID: 22318836; PMCID: PMC3282950.
  • Clinical and Laboratory Standards Institute (CLSI) (2021) Perfor- mance Standards for Antimicrobial. Document M100-Ed.31., Wayne, PA: CLSI.
  • Del Pozo J. L. (2018). Biofilm-related disease. Expert Review of Anti- Infective Therapy, 16(1), 51–65. https://doi.org/10.1080/14787210.2018.1417036
  • Dosler, S. & Karaaslan E. (2014). Inhibition and destruction of Pseu- domonas aeruginosa biofilms by antibiotics and antimicrobial peptides. Peptides, 62, 32-37.
  • Dumaru, R., Baral, R., & Shrestha, L. B. (2019). Study of biofilm for- mation and antibiotic resistance pattern of gram-negative Ba- cilli among the clinical isolates at BPKIHS, Dharan. BMC Research Notes, 12(1), 38. https://doi.org/10.1186/s13104-019-4084-8
  • Eichenberger, E. M., & Thaden, J. T. (2019). Epidemiology and mechanisms of resistance of extensively drug resistant Gram- negative bacteria. Antibiotics, 8(2), 37.
  • El-Gamal, M. I., Brahim, I., Hisham, N., Aladdin, R., Mohammed, H., & Bahaaeldin, A. (2017). Recent updates of carbapenem antibiot- ics. European Journal of Medicinal Chemistry, 131, 185–195. https:// doi.org/10.1016/j.ejmech.2017.03.022
  • Hammoudi Halat, D., & Ayoub Moubareck, C. (2020). The current burden of carbapenemases: Review of significant properties and dissemination among Gram-negative bacteria. Antibiotics, 9(4), 186.
  • İnce, G., Mirza, H. C., Üsküdar Güçlü, A., Gümüş, H., Erol, Ç., & Başustaoğlu, A. (2021). Comparison of in vitro activities of plazo- micin and other aminoglycosides against clinical isolates of Kleb- siella pneumoniae and Escherichia coli. The Journal of Antimicrobial Chemotherapy, 76(12), 3192–3196. https://doi.org/10.1093/jac/ dkab331
  • Jamal, M., Ahmad, W., Andleeb, S., Jalil, F., Imran, M., Nawaz, M. A., Hussain, T., Ali, M., Rafiq, M., & Kamil, M. A. (2018). Bacterial biofilm and associated infections. Journal of the Chinese Medical Associa- tion: JCMA, 81(1), 7–11. https://doi.org/10.1016/j.jcma.2017.07.012
  • Jia, X., Ma, W., He, J., Tian, X., Liu, H., Zou, H., & Cheng, S. (2020).
  • Heteroresistance to cefepime in Pseudomonas aeruginosa bacter- aemia. International Journal of Antimicrobial Agents, 55(3), 105832. https://doi.org/10.1016/j.ijantimicag.2019.10.013
  • Magiorakos, A. P., Srinivasan, A., Carey, R. B., Carmeli, Y., Falagas, M. E., Giske, C. G., & Monnet, D. L. (2012). Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clinical Microbiology and Infection, 18(3), 268- 281.
  • Malone, L., & Kwon, D. H. (2013). Carbapenem-associated mul- tidrug-resistant Acinetobacter baumannii are sensitised by az- treonam in combination with polyamines. International journal of Antimicrobial Agents, 41(1), 70–74. https://doi.org/10.1016/j. ijantimicag.2012.08.009
  • Mirza, H. C., Sancak, B., (2020). Comparison of antimicrobial sus- ceptibilities of Escherichia coli isolated from urinary cultures of different patient groups: A university hospital experience. Klimik Dergisi, 33(3), 270-6.
  • Mojica, M. F., Humphries, R., Lipuma, J. J., Mathers, A. J., Rao, G. G., Shelburne, S. A., Fouts, D. E., Van Duin, D., & Bonomo, R. A. (2022). Clinical challenges treating Stenotrophomonas maltophilia in- fections: an update. JAC-Antimicrobial Resistance, 4(3), dlac040. https://doi.org/10.1093/jacamr/dlac040
  • Namaei, M. H., Yousefi, M., Askari, P., Roshanravan, B., Hashemi, A., & Rezaei, Y. (2021). High prevalence of multidrug-resistant non-fermentative Gram-negative bacilli harboring blaIMP-1 and blaVIM-1 metallo-beta-lactamase genes in Birjand, south-east Iran. Iranian Journal of Microbiology, 13(4), 470–479. https://doi. org/10.18502/ijm.v13i4.6971
  • Nirwati, H., Sinanjung, K., Fahrunissa, F., Wijaya, F., Napitupulu, S., Hati, V. P., Hakim, M. S., Meliala, A., Aman A. T., & Nuryastuti, T. (2019). Biofilm formation and antibiotic resistance of Klebsiella pneumoniae isolated from clinical samples in a tertiary care hos- pital, Klaten, Indonesia. BMC Proceedings, 13(11), 1-8. https://doi. org/10.1186/s12919-019-0176-7
  • Pang, Z., Raudonis, R., Glick, B. R., Lin, T. J., & Cheng, Z. (2019). An- tibiotic resistance in Pseudomonas aeruginosa: mechanisms and alternative therapeutic strategies. Biotechnology Advances, 37(1), 177–192. https://doi.org/10.1016/j.biotechadv.2018.11.013
  • Peeters, E., Nelis, H. J. & Coenye. T., (2008). Comparison of mul- tiple methods for quantification of microbial biofilms grown in microtiter plates. Journal of Microbiology Methods, 72(2), 157-165. doi:10.1016/j.mimet.2007.11.010
  • Perdikouri, E. I. A., Arvaniti, K., Lathyris, D., Apostolidou Kiouti, F., Siskou, E., Haidich, A. B., & Papandreou, C. (2019). Infections due to multidrug-resistant bacteria in oncological patients: insights from a five-year epidemiological and clinical analysis. Microorgan- isms, 7(9), 277.
  • Peters, L., Olson, L., Khu, D. T., Linnros, S., Le, N. K., Hanberger, H. & Larsson, M. (2019). Multiple antibiotic resistance as a risk factor for mortality and prolonged hospital stay: a cohort study among neonatal intensive care patients with hospital-acquired infec- tions caused by Gram-negative bacteria in Vietnam. PloS one, 14(5), e0215666.
  • Rice, L. B. (2008). Federal funding for the study of antimicrobial resistance in nosocomial pathogens: no ESKAPE. The Journal of Infectious Diseases, 197(8), 1079-1081.
  • Thaden, J. T., Li, Y., Ruffin, F., Maskarinec, S. A., Hill-Rorie, J. M., Wan- da, L. C., & Fowler Jr, V. G. (2017). Increased costs associated with bloodstream infections caused by multidrug-resistant gram-neg- ative bacteria are due primarily to patients with hospital-acquired infections. Antimicrobial Agents and Chemotherapy, 61(3), e01709-16.
  • Vivas, R., Barbosa, A. A. T., Dolabela, S. S., & Jain, S. (2019). Multi- drug-resistant bacteria and alternative methods to control them: an overview. Microbial Drug Resistance, 25(6), 890-908.
  • Wang, G., Zhao, G., Chao, X., Xie, L., & Wang, H. (2020). The charac- teristic of virulence, biofilm and antibiotic resistance of Klebsiella pneumoniae. International Journal of Environmental Research and Public Health, 17(17), 6278.
  • World Health Organization (WHO) https://www.who.int/news- room/fact-sheets/detail/antimicrobial-resistance
  • Yadav, S.K., Bhujel, R. & Mishra, S.K. (2020). Emergence of multi- drug-resistant non-fermentative gram negative bacterial infec- tion in hospitalized patients in a tertiary care center of Nepal. BMC Res Notes, 13, 319. https://doi.org/10.1186/s13104-020-05163-6
There are 34 citations in total.

Details

Primary Language English
Subjects Pharmacology and Pharmaceutical Sciences
Journal Section Original Article
Authors

Özlem Oyardı 0000-0001-9992-7225

Mayram Tuysuz 0000-0003-0823-631X

Fatıma Nur Yılmaz 0000-0001-8442-8538

Nese Inan 0000-0002-1559-6244

Ayşe Seher Birteksöz Tan 0000-0001-8767-710X

Project Number 37825
Publication Date April 28, 2023
Submission Date August 7, 2022
Published in Issue Year 2023

Cite

APA Oyardı, Ö., Tuysuz, M., Yılmaz, F. N., Inan, N., et al. (2023). Antibiotic susceptibility and biofilm formation of multi-drug resistant Gram-negative bacteria. İstanbul Journal of Pharmacy, 53(1), 45-50. https://doi.org/10.26650/IstanbulJPharm.2023.1156339
AMA Oyardı Ö, Tuysuz M, Yılmaz FN, Inan N, Birteksöz Tan AS. Antibiotic susceptibility and biofilm formation of multi-drug resistant Gram-negative bacteria. iujp. April 2023;53(1):45-50. doi:10.26650/IstanbulJPharm.2023.1156339
Chicago Oyardı, Özlem, Mayram Tuysuz, Fatıma Nur Yılmaz, Nese Inan, and Ayşe Seher Birteksöz Tan. “Antibiotic Susceptibility and Biofilm Formation of Multi-Drug Resistant Gram-Negative Bacteria”. İstanbul Journal of Pharmacy 53, no. 1 (April 2023): 45-50. https://doi.org/10.26650/IstanbulJPharm.2023.1156339.
EndNote Oyardı Ö, Tuysuz M, Yılmaz FN, Inan N, Birteksöz Tan AS (April 1, 2023) Antibiotic susceptibility and biofilm formation of multi-drug resistant Gram-negative bacteria. İstanbul Journal of Pharmacy 53 1 45–50.
IEEE Ö. Oyardı, M. Tuysuz, F. N. Yılmaz, N. Inan, and A. S. Birteksöz Tan, “Antibiotic susceptibility and biofilm formation of multi-drug resistant Gram-negative bacteria”, iujp, vol. 53, no. 1, pp. 45–50, 2023, doi: 10.26650/IstanbulJPharm.2023.1156339.
ISNAD Oyardı, Özlem et al. “Antibiotic Susceptibility and Biofilm Formation of Multi-Drug Resistant Gram-Negative Bacteria”. İstanbul Journal of Pharmacy 53/1 (April 2023), 45-50. https://doi.org/10.26650/IstanbulJPharm.2023.1156339.
JAMA Oyardı Ö, Tuysuz M, Yılmaz FN, Inan N, Birteksöz Tan AS. Antibiotic susceptibility and biofilm formation of multi-drug resistant Gram-negative bacteria. iujp. 2023;53:45–50.
MLA Oyardı, Özlem et al. “Antibiotic Susceptibility and Biofilm Formation of Multi-Drug Resistant Gram-Negative Bacteria”. İstanbul Journal of Pharmacy, vol. 53, no. 1, 2023, pp. 45-50, doi:10.26650/IstanbulJPharm.2023.1156339.
Vancouver Oyardı Ö, Tuysuz M, Yılmaz FN, Inan N, Birteksöz Tan AS. Antibiotic susceptibility and biofilm formation of multi-drug resistant Gram-negative bacteria. iujp. 2023;53(1):45-50.