Multidrug Resistance Patterns of Escherichia coli from Wastewater in Baghdad

Wid Maan AL-Bayati; Hiba  Qasim Hameed

doi:10.30526/39.4.4351

Authors

Wid Maan AL-Bayati Department of Biology, College of Education Pure Sciences (Ibn Al-Haitham), University of Baghdad, Baghdad, Iraq https://orcid.org/0009-0001-3230-4442
Hiba Qasim Hameed Department of Biology, College of Education for Pure Science (Ibn Al-Haitham), University of Baghdad, Baghdad, Iraq https://orcid.org/0000-0002-3292-8837

DOI:

https://doi.org/10.30526/39.4.4351

Keywords:

Escherichia coli, hospital wastewater, antibiotic resistance

Abstract

Hospital wastewater is believed to be a leading source of antibiotic resistant bacteria, which presents a risk to both the ecosystem and public health. The current study endeavored to isolate and identify Escherichia coli from the wastewater of multiple hospitals in Baghdad City, Iraq, and characterize its antibiotic resistance patterns. The study resulted in identifying, by biochemical means using the VITEK system, a total of 50 E.coli isolates. Antibiotic susceptibility tests for 11 antibiotics were conducted on each isolate via the Kirby–Bauer (KB) disk diffusion method. Overall, all isolates showed considerable resistance to the following antibiotics: erythromycin and clarithromycin (100%), amoxicillin (86%), cefoxitin (72%), ceftriaxone (66%), and tetracycline (56%). Conversely, gentamicin (6%) and meropenem (16%) exhibited the lowest resistance patterns among all isolates. The Multiple Antibiotic Resistance (MAR) index indicated that 44 isolates (88%) exceeded 0.2, signifying that a high number of isolates were contaminated or exposed to antibiotics

Author Biographies

Wid Maan AL-Bayati, Department of Biology, College of Education Pure Sciences (Ibn Al-Haitham), University of Baghdad, Baghdad, Iraq

-
Hiba Qasim Hameed, Department of Biology, College of Education for Pure Science (Ibn Al-Haitham), University of Baghdad, Baghdad, Iraq

-

References

1. Amin M, Al-Mohannadi A, Al-Ansari M, Hassan M. Wastewater from hospitals in developing countries: occurrence, risks, and challenges of treatment technologies. Appl Microbiol. 2024;3,60:1–12.

2. Rizzo L, Manaia C, Merlin C, Schwartz T, Dagot C, Ploy MC, Michael I, Fatta-Kassinos D. Urban wastewater treatment plants as hotspots for antibiotic resistant bacteria and genes spread into the environment: a review. Sci Total Environ. 2013;447:345-60. https://doi.org/10.1016/j.scitotenv.2013.01.032

3. Pandey A, Afsheen AF, Tiwari SK. Isolation and characterization of multidrug resistant cultures from wastewater. J Pharm Biomod Sci. 2011;13(7): 1-7.

4. Anitha J, Jayraaj IA. Isolation and identification of bacteria in biomedical wastes. Int J Pharm Pharm Sci. 2012;4(5):386–388.

5. Shahzad A, Mian AH, Ul Haq I, Khan MA, Matiullah, Ali K, Hamid T. The emergence of different bacterial pathogens in hospital wastewater samples and their antibiotic resistance pattern. Mater Circ Econ. 2021;3:28. https://doi.org/10.1007/s42824-021-00046-y

6. Zhang S, Wang Y, Song H, Lu J, Yuan Z, Guo J. Dissemination of antibiotic resistance genes in aquatic environments: a review. Sci Total Environ. 2018;627:1627–1639.

7. Duarte AC, Rodrigues S, Afonso A, Nogueira A, Coutinho P. Antibiotic resistance in drinking water: old and new strategies to remove antibiotics, resistant bacteria, and resistance genes. Pharmaceuticals. 2022;15(4):393. https://doi.org/10.3390/ph15040393.

8. Obaid WA. Comprehensive phenotypic and genotypic analysis of multidrug-resistant and virulent Enterococcus spp. from untreated hospital wastewater and river water in Baghdad, Iraq. Res Sq. 2025; preprint. https://doi.org/10.21203/rs.3.rs-7649505/v1

9. Naidoo N, Zishiri OT. Presence, pathogenicity, antibiotic resistance, and virulence factors of Escherichia coli: a review. Bacteria. 2025;4:16. https://doi.org/10.3390/bacteria4010016

10. Hamady DR, Ibrahim SK. The study on ability of Escherichia coli isolated from different clinical cases to biofilm formation and detection of csgD gene responsible for curli production. Biochem Cell Arch. 2020;20(2):5553–5557.

11. Zhang C, Qiu S, Wang Y, Hao R, Li X, Shi Y, Hu X, An D, Li Z. Higher isolation of NDM-1 producing Acinetobacter baumannii from hospital sewage in Beijing. PLoS One. 2013;8:e64857. https://doi.org/10.1371/journal.pone.0064857.

12. Abdulsattar B, Abdulsattar JO, Habeeb K. Study of antimicrobial resistance pattern of Escherichia coli and Klebsiella strains and multivariate analysis for water quality assessment of Tigris River, Baghdad, Iraq. Nat Environ Pollut Technol. 2020;19(3):1051–1058. https://doi.org/10.46488/nept.2020.v19i03.050.

13. Mahdi ZM, Al-Saadi HM, Al-Mayali HK. Detection of resistance genes in waterborne Escherichia coli isolates from Baghdad wastewater. J Water Health. 2024;22(1):55–66.

14. Brooks GF, Carroll KC, Butel JS, Morse SA, Mietzner TA. Jawetz, Melnick & Adelberg’s Medical Microbiology. 26th ed. New York: McGraw-Hill; 2013. p.245–248.

15. Tille PM. Bailey & Scott’s Diagnostic Microbiology. 14th ed. St. Louis (MO): Elsevier; 2017. 1115 p.

16. Hudzicki J. Kirby–Bauer disk diffusion susceptibility test protocol. Am Soc Microbiol. 2009.

17. Clinical and Laboratory Standards Institute. M100: Performance standards for antimicrobial susceptibility testing. 35th ed. Wayne (PA): Clin Lab Stand Inst.; 2025.

18. Isaiah DO, Otokunefor K, Agbagwa OE. Multiple antibiotic resistance indexing and molecular identification of Escherichia coli isolated from clinical and nonclinical sources in Port Harcourt Metropolis, Nigeria. Pan Afr Med J. 2025;51:11. https://doi.org/10.11604/pamj.2025.51.11.38524.

19. Joshi DC, Prasad S, Sharma A, Joshi N. Pathological contaminants in wastewater from hospital waste: implications and mitigation strategies. In: Emerging Contaminants in Water and Wastewater: Sources and Substances. Cham: Springer Nature Switzerland; 2025. p. 43-85. https://doi.org/10.1007/978-3-031-82579-8_3.

20. Manaia CM, Rocha J, Scaccia N, Marano R, Radu E, Biancullo F, Cerqueira F, Fortunato G, Iakovides IC, Zammit I, Kampouris I. Antibiotic resistance in wastewater treatment plants: Tackling the black box. Environ Int. 2018;115:312-24. https://doi.org/10.1016/j.envint.2018.03.044

21. Altameemi Z, Talib A, Fakhry S. Detection of antibiotic-resistant Escherichia coli in the Euphrates River, Iraq. bioRxiv [Preprint]. 2025. https://doi.org/10.1101/2025.08.30.673253

22. Kumar G, Balakrishana K, Mukhopadhyay C, Eshwara VK, Kadam SB. Characterization and comparative analysis of antimicrobial resistance in Escherichia coli from hospital and municipal wastewater treatment plants. J Water Health. 2024;22(12):2274–2284. https://doi.org/10.2166/wh.2024.126.

23. Gaspar CM, Cziszter LT, Lăzărescu C, Tîrban I, Pentea M, Butnariu M. Antibiotic resistance among Escherichia coli isolates from hospital wastewater compared to community wastewater. Water. 2021;13(24):3449. https://doi.org/10.3390/w13233449.

24. Dagher LA, Joeman D, Kharroubi SH, Jaafer H, Kasem I. Nationwide assessment of water quality in rivers across Lebanon by quantifying fecal indicator densities and profiling antibiotic resistance of Escherichia coli. Antibiotics. 2021;10(9):1083. https://doi.org/10.3390/antibiotics10091083

25. Wu D, Ding Y, Yao K, Gao W, Wang Y. Antimicrobial resistance analysis of clinical Escherichia coli isolates in neonatal ward. Front Pediatrics. 2021;9:670470. https://doi.org/10.3389/fped.2021.670470.

26. Sadeq HS, Waheeb MQ. Phenotypic characterization of Escherichia coli and determination of their sensitivity to antibiotics. Microb Sci Arch. 2024;4(3):17–23. https://doi.org/10.47587/msa.2024.4302.

27. Karumangnye P, Rugaika A, Mlei K, Machunda R. Antibiotic resistance patterns of Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa isolated from hospital wastewater. Appl Microbiol. 2023;3(2):867–882. https://doi.org/10.3390/applmicrobiol3030060.

28. Al-Qatan MAR, Al-Khayyat MZ, Saeed HK. Isolation of antibiotic-resistant bacteria from sewage water in Mosul City. Biochem Cell Arch. 2019;19(1):1415–1418.

29. Tufa KF, Birhanu AG. Antimicrobial resistance profile of Escherichia coli isolated from hospital and industrial wastewater systems. Environ Health Insights. 2025;19:11786302251339254.‏ https://doi.org/10.1177/11786302251339254.

30. Venter H, Mowla R, Ohene-Agyei T, Ma S. RND-type drug efflux pumps from Gram-negative bacteria: molecular mechanism and inhibition. Front Microbiol. 2015;6:377. https://doi.org/10.3389/fmicb.2015.00377.

31. Khattab A, Nizam DA, Alamir DL. Isolation of Escherichia coli from effluents of some Damascus University hospitals and determination of their susceptibility to beta-lactam antibiotics. Damascus Univ J Basic Sci. 2023;39(3):112–126.

32. Davidova-Gerzova L, Lausova J, Sukkar I, Nesporova K, Nechutna L, Vikova K, Svobodova M, Soukupova M, Palenikova J, Kaspar J, Dotejska M. Hospital and community wastewater as a source of multidrug-resistant ESBL-producing Escherichia coli. Front Cell Infect Microbiol. 2023;13:1084085. https://doi.org/10.3389/fcimb.2023.1184081.

33. El Garraoui Q, Logman A, Amouat FE, Hasnaou S, Soras N, Logman S. Antimicrobial resistance and molecular detection of extended-spectrum β-lactamase–producing Escherichia coli in municipal wastewater in Marrakech. PLoS One. 2025;20(6):e0325816. https://doi.org/10.1371/journal.pone.0325816

34. Sader HS, Mendes RE, Arends SR, Carvalhaes CG, Castanheira M. Antimicrobial activities of aztreonam-avibactam and comparator agents tested against Enterobacterales from European hospitals analysed by geographic region and infection type (2019–2020). Europ J Clin Microbiol Infect Dis. 2022;41(3):477-87. https://doi.org/10.1007/s10096-022-04400-z.

35. Abdelgaleil RR, Ibrahim HA, Ahmed DS, Ahmed AB. Multidrug-resistant Escherichia coli in wastewater sources: a comparative study and identification of resistance hotspots. BMC Microbiol. 2023;23:298. https://doi.org/10.1186/s12866-025-04244-5.

36. Amador PP, Fernandes RM, Prudêncio MC, Barreto MP, Duarte IM. Antibiotic resistance in wastewater: occurrence and fate of Enterobacteriaceae producers of class A and class C β-lactamases. J Environ Sci Health A. 2015;50(1):26–39. https://doi.org/10.1080/10934529.2015.964602

37. Endalamaw K, Tadesse S, Asmare Z, Rebede D, Erkihun M, Abela B. Antimicrobial resistance profile of bacteria from hospital wastewater at two specialized hospitals in Bahir Dar City, Ethiopia. BMC Microbiol. 2024;24(1):525. https://doi.org/10.1186/s12866-024-03992-8

38. Mataseje LF, Niu YD, Tyler SD, Daignault S, Zhand T, Low DE. Characterization of cefoxitin-resistant Escherichia coli isolates from recreational beaches and private drinking water in Canada between 2004 and 2006. Antimicrob Agents Chemother. 2009;53(7):3126–3130. https://doi.org/10.1128/AAC.01330-08

39. Seguni NZ, Kimera ZI, Msafiri F, Mgaya FX, Joachim A, Mwingwa A, Matee MI. Multidrug-resistant Escherichia coli and Klebsiella pneumoniae isolated from hospital sewage flowing through community sewage system and discharging into the Indian Ocean. Bulletin of the National Research Centre. 2023 May 11;47(1):66.‏ https://doi.org/10.1186/s42269-023-01039-4.

40. Hussein IA, Talib AA, Fakhry SS. Prevalence and antimicrobial susceptibility of Escherichia coli and Klebsiella pneumoniae isolated from the Tigris River, Baghdad, Iraq. Iraqi J Sci. 2025;66(12): 5408–5423. https://doi.org/10.24996/ijs.2025.66.12.%g.

41. Adegoke AA, Madu CE, Aiyegoro OA, Stenström TA, Okoh AI. Antibiogram and beta-lactamase genes among cefotaxime resistant E. coli from wastewater treatment plant. Antimicrob Resist Infect Cont. 2020;9(1):46.‏ https://doi.org/10.1186/s13756-020-0702-4