Articles In Press                   Back to the articles list | Back to browse issues page

XML Print


1- Medical Microbiology Research Center, Qazvin University of Medical Sciences, Qazvin, Iran
2- Agronomy department, Faculty of Agriculture, Takestan Branch, Islamic Azad University
3- Medical Microbiology Research Center, Qazvin University of Medical Sciences, Qazvin, Iran , farhadnikkhahi@gmail.com
Abstract:   (131 Views)

Background and Objectives: Avian pathogenic Escherichia coli (APEC) is a major cause of colibacillosis in poultry, contributing to systemic disease and economic loss. This study investigated virulence gene profiles, biofilm formation, and antimicrobial and disinfectant resistance in E. coli isolates from broiler chickens in Qazvin, Iran.
Methods: Liver samples were collected from 50 broiler chickens diagnosed with colibacillosis. E. coli isolates were confirmed biochemically. Antibiotic susceptibility was evaluated using the disk diffusion method, and virulence genes were detected via PCR. Biofilm formation was assessed using crystal violet staining. MIC and MBC values for formaldehyde, glutaraldehyde, and hydrogen peroxide were determined using broth microdilution.
Results: Among 50 isolates, 82% exhibited multidrug resistance (MDR) and 30% were ESBL producers. The most frequent gene was iss (66%), and papC was the least common (4%). All isolates formed biofilms, with 72% classified as strong producers. Formaldehyde and hydrogen peroxide exhibited the lowest MIC and MBC values (≤ 0.009%), while glutaraldehyde required higher concentrations for inhibitory and bactericidal activity (0.078%).
Conclusion: High rates of MDR and biofilm formation among APEC isolates highlight the need for effective antimicrobial stewardship and disinfection strategies in poultry production systems.

     
Type of Study: Original Research Article | Subject: Medical Bacteriology
Received: 2025/05/25 | Accepted: 2025/08/10 | ePublished: 2025/08/18

References
1. Sivaranjani M, McCarthy MC, Sniatynski MK, Wu L, Dillon J-AR, Rubin JE, et al. Biofilm formation and antimicrobial susceptibility of E. coli associated with colibacillosis outbreaks in broiler chickens from Saskatchewan. Front Microbiol. 2022;13:841516. [DOI:10.3389/fmicb.2022.841516] [PMID] [PMCID]
2. de Brito BG, Gaziri LCJ, Vidotto MC. Virulence factors and clonal relationships among Escherichia coli strains isolated from broiler chickens with cellulitis. Infect Immun. 2003;71(7):4175-7. [DOI:10.1128/IAI.71.7.4175-4177.2003] [PMID] [PMCID]
3. Kathayat D, Lokesh D, Ranjit S, Rajashekara G. Avian pathogenic Escherichia coli (APEC): an overview of virulence and pathogenesis factors, zoonotic potential, and control strategies. Pathogens. 2021;10(4):467. [DOI:10.3390/pathogens10040467] [PMID] [PMCID]
4. Joseph J, Zhang L, Adhikari P, Evans JD, Ramachandran R. Avian pathogenic Escherichia coli (APEC) in broiler breeders: an overview. Pathogens. 2023;12(11):1280. [DOI:10.3390/pathogens12111280] [PMID] [PMCID]
5. Sarowska J, Futoma-Koloch B, Jama-Kmiecik A, Frej-Madrzak M, Ksiazczyk M, Bugla-Ploskonska G, et al. Virulence factors, prevalence and potential transmission of extraintestinal pathogenic Escherichia coli isolated from different sources: recent reports. Gut Pathog. 2019;11:1-16. [DOI:10.1186/s13099-019-0290-0] [PMID] [PMCID]
6. Paixao A, Ferreira A, Fontes M, Themudo P, Albuquerque T, Soares M, et al. Detection of virulence-associated genes in pathogenic and commensal avian Escherichia coli isolates. Poult Sci. 2016;95(7):1646-52. [DOI:10.3382/ps/pew087] [PMID]
7. Ovi F, Zhang L, Nabors H, Jia L, Adhikari P. A compilation of virulence-associated genes that are frequently reported in avian pathogenic Escherichia coli (APEC) compared to other E. coli. J Appl Microbiol. 2023;134(3):lxad014. [DOI:10.1093/jambio/lxad014] [PMID]
8. Gao Q, Jia X, Wang X, Xiong L, Gao S, Liu X. The avian pathogenic Escherichia coli O2 strain E058 carrying the defined aerobactin-defective iucD or iucDiutA mutation is less virulent in the chicken. Infect Genet Evol. 2015;30:267-77. [DOI:10.1016/j.meegid.2014.12.038] [PMID]
9. Lynne AM, Foley SL, Nolan LK. Immune response to recombinant Escherichia coli Iss protein in poultry. Avian Dis. 2006;50(2):273-6. [DOI:10.1637/7441-092105R.1] [PMID]
10. Aleksandrowicz A, Khan MM, Sidorczuk K, Noszka M, Kolenda R. Whatever makes them stick-Adhesins of avian pathogenic Escherichia coli. Vet Microbiol. 2021;257:109095. [DOI:10.1016/j.vetmic.2021.109095] [PMID]
11. Maluta RP, Gatti MSV, Joazeiro PP, De Paiva JB, Rojas TCG, Silveira F, et al. Avian extraintestinal Escherichia coli exhibits enterotoxigenic-like activity in the in vivo rabbit ligated ileal loop assay. Foodborne Pathog Dis. 2014;11(6):484-9. [DOI:10.1089/fpd.2013.1719] [PMID]
12. Gérard F, Pradel N, Wu L-F. Bactericidal activity of colicin V is mediated by an inner membrane protein, SdaC, of Escherichia coli. J Bacteriol. 2005;187(6):1945-50. [DOI:10.1128/JB.187.6.1945-1950.2005] [PMID] [PMCID]
13. Rodrigues SV, Laviniki V, Borges KA, Furian TQ, Moraes HL, Nascimento VP, et al. Biofilm formation by avian pathogenic Escherichia coli is not related to in vivo pathogenicity. Curr Microbiol. 2019;76:194-9. [DOI:10.1007/s00284-018-1608-8] [PMID]
14. Ghamari M, Sabzi S, Rajabi E, Khodadadi G, Navidifar T, Sadeghi Z, et al. Probiotics, Prebiotics, Synbiotics, Postbiotics, and Bioactive Agents in Modulating Harmful Oral Biofilms. Probiotics Antimicrob Proteins. 2025:1-32. [DOI:10.1007/s12602-025-10636-w] [PMID]
15. Uruén C, Chopo-Escuin G, Tommassen J, Mainar-Jaime RC, Arenas J. Biofilms as promoters of bacterial antibiotic resistance and tolerance. Antibiotics. 2020;10(1):3. [DOI:10.3390/antibiotics10010003] [PMID] [PMCID]
16. Roy R, Tiwari M, Donelli G, Tiwari V. Strategies for combating bacterial biofilms: A focus on anti-biofilm agents and their mechanisms of action. Virulence. 2018;9(1):522-54. [DOI:10.1080/21505594.2017.1313372] [PMID] [PMCID]
17. Gržinić G, Piotrowicz-Cieślak A, Klimkowicz-Pawlas A, Górny RL, Ławniczek-Wałczyk A, Piechowicz L, et al. Intensive poultry farming: A review of the impact on the environment and human health. Sci Total Environ. 2023;858:160014. [DOI:10.1016/j.scitotenv.2022.160014] [PMID]
18. Mohammadzadeh R, Shahbazi S, Khodaei N, Sabzi S. Emerging Therapeutic Strategies to Combat Antimicrobial Resistance in the Post‐Antibiotic Era. J Basic Microbiol. 2025:e70070. [DOI:10.1002/jobm.70070] [PMID]
19. Shahkolahi S, Shakibnia P, Shahbazi S, Sabzi S, Badmasti F, Asadi Karam MR, et al. Detection of ESBL and AmpC producing Klebsiella pneumoniae ST11 and ST147 from urinary tract infections in Iran. Acta Microbiol Immunol Hung. 2022;69(4):303-13. [DOI:10.1556/030.2022.01808] [PMID]
20. Al-Marri T, Al-Marri A, Al-Zanbaqi R, Al Ajmi A, Fayez M. Multidrug resistance, biofilm formation, and virulence genes of Escherichia coli from backyard poultry farms. Vet World. 2021;14(11):2869. [DOI:10.14202/vetworld.2021.2869-2877] [PMID] [PMCID]
21. Gharieb R, Saad M, Khedr M, El Gohary A, Ibrahim H. Occurrence, virulence, carbapenem resistance, susceptibility to disinfectants and public health hazard of Pseudomonas aeruginosa isolated from animals, humans and environment in intensive farms. J Appl Microbiol. 2022;132(1):256-67. [DOI:10.1111/jam.15191] [PMID]
22. Soumet C, Méheust D, Pissavin C, Le Grandois P, Fremaux B, Feurer C, et al. Reduced susceptibilities to biocides and resistance to antibiotics in food‐associated bacteria following exposure to quaternary ammonium compounds. J Appl Microbiol. 2016;121(5):1275-81. [DOI:10.1111/jam.13247] [PMID]
23. Nikkhahi F, Robatjazi S, Niazadeh M, Javadi A, Shahbazi G, Aris P, et al. First detection of mobilized colistin resistance mcr-1 gene in Escherichia coli isolated from livestock and sewage in Iran. New Microbes New Infect. 2021;41:100862. [DOI:10.1016/j.nmni.2021.100862] [PMID] [PMCID]
24. Ahmed OB, Dablool AS. Quality improvement of the DNA extracted by boiling method in gram negative bacteria. Int J Bioassays. 2017;6(4):5347-9. [DOI:10.21746/ijbio.2017.04.004]
25. Anari RK, Nikkhahi F, Javadi A, Bakht M, Rostamani M, Kelishomi FZ, et al. Evaluation of antibacterial activity of five biocides and the synergistic effect of biocide/EDTA combinations on biofilm-producing and non-producing Stenotrophomonas maltophilia strains isolated from clinical specimens in Iran. BMC Microbiol. 2022;22(1):257. [DOI:10.1186/s12866-022-02664-1] [PMID] [PMCID]
26. Stepanović S, Ćirković I, Mijač V, Švabić-Vlahović M. Influence of the incubation temperature, atmosphere and dynamic conditions on biofilm formation by Salmonella spp. Food Microbiol. 2003;20(3):339-43. [DOI:10.1016/S0740-0020(02)00123-5]
27. Thapa D, Chapagain A. Antibiogram of Escherichia coli isolated from avian colibacillosis in Chitwan district of Nepal. Int J Appl Sci Biotechnol. 2020;8(1):52-60. [DOI:10.3126/ijasbt.v8i1.28254]
28. Subedi M, Luitel H, Devkota B, Bhattarai RK, Phuyal S, Panthi P, et al. Antibiotic resistance pattern and virulence genes content in avian pathogenic Escherichia coli (APEC) from broiler chickens in Chitwan, Nepal. BMC Vet Res. 2018;14:1-6. https://doi.org/10.1186/s12917-018-1442-z [DOI:10.1186/s12917-018-1453-9] [PMID] [PMCID]
29. Ameen-Ur-Rashid SS, Khan M, Rafiullah AA, Anwar M. Isolation of Escherichia coli from Poultry Liver and its Antibiogram Profile. Res J Vet Pract. 2017;5(1):12-4.
30. Pokharel S, Raut S, Adhikari B. Tackling antimicrobial resistance in low-income and middle-income countries. BMJ Glob Health. 2019;4(6):e002104. [DOI:10.1136/bmjgh-2019-002104] [PMID] [PMCID]
31. Van Boeckel TP, Brower C, Gilbert M, Grenfell BT, Levin SA, Robinson TP, et al. Global trends in antimicrobial use in food animals. Proc Natl Acad Sci. 2015;112(18):5649-54. [DOI:10.1073/pnas.1503141112] [PMID] [PMCID]
32. Khong M, Snyder A, Magnaterra A, Young M, Barbieri N, Weimer S. Antimicrobial resistance profile of Escherichia coli isolated from poultry litter. Poult Sci. 2023;102(1):102305. [DOI:10.1016/j.psj.2022.102305] [PMID] [PMCID]
33. Tohmaz M, Askari Badouei M, Kalateh Rahmani H, Hashemi Tabar G. Antimicrobial resistance, virulence associated genes and phylogenetic background versus plasmid replicon types: the possible associations in avian pathogenic Escherichia coli (APEC). BMC Vet Res. 2022;18(1):421. [DOI:10.1186/s12917-022-03496-x] [PMID] [PMCID]
34. Ahmed AM, Shimamoto T, Shimamoto T. Molecular characterization of multidrug-resistant avian pathogenic Escherichia coli isolated from septicemic broilers. Int J Med Microbiol. 2013;303(8):475-83. [DOI:10.1016/j.ijmm.2013.06.009] [PMID]
35. Dolejska M, Duskova E, Rybarikova J, Janoszowska D, Roubalova E, Dibdakova K, et al. Plasmids carrying bla CTX-M-1 and qnr genes in Escherichia coli isolates from an equine clinic and a horseback riding centre. J Antimicrob Chemother. 2011;66(4):757-64. [DOI:10.1093/jac/dkq500] [PMID]
36. Roth N, Käsbohrer A, Mayrhofer S, Zitz U, Hofacre C, Domig KJ. The application of antibiotics in broiler production and the resulting antibiotic resistance in Escherichia coli: A global overview. Poult Sci. 2019;98(4):1791-804. [DOI:10.3382/ps/pey539] [PMID] [PMCID]
37. Akbari P, Asadpour L. An Evaluation of Antimicrobial Resistance and Virulence Potential of Escherichia coli Obtained from Feces of Ornamental Birds in Guilan, Iran. Iran J Med Microbiol. 2022;16(5):405-11. [DOI:10.30699/ijmm.16.5.405]
38. Chen X, Zhang W, Yin J, Zhang N, Geng S, Zhou X, et al. Escherichia coli isolates from sick chickens in China: changes in antimicrobial resistance between 1993 and 2013. Vet J. 2014;202(1):112-5. [DOI:10.1016/j.tvjl.2014.06.016] [PMID]
39. Tongkamsai S, Nakbubpa K. Extended-spectrum beta-lactamase (ESBL) production and virulence genes profile of avian pathogenic Escherichia coli (APEC) isolated from broiler chickens in eastern Thailand: https://doi. org/10.12982/VIS. 2024.016. Vet Integr Sci. 2024;22(1):207-18. [DOI:10.12982/VIS.2024.016]
40. Dierikx C, van der Goot J, Fabri T, van Essen-Zandbergen A, Smith H, Mevius D. Extended-spectrum-β-lactamase-and AmpC-β-lactamase-producing Escherichia coli in Dutch broilers and broiler farmers. J Antimicrob Chemother. 2013;68(1):60-7. [DOI:10.1093/jac/dks349] [PMID]
41. Robinson TP, Bu D, Carrique-Mas J, Fèvre EM, Gilbert M, Grace D, et al. Antibiotic resistance is the quintessential One Health issue. Trans R Soc Trop Med Hyg. 2016;110(7):377-80. [DOI:10.1093/trstmh/trw048] [PMID] [PMCID]
42. Mondal AH, Khare K, Saxena P, Debnath P, Mukhopadhyay K, Yadav D. A review on colistin resistance: an antibiotic of last resort. Microorganisms. 2024;12(4):772. [DOI:10.3390/microorganisms12040772] [PMID] [PMCID]
43. Cunha MPV, de Oliveira MGX, de Oliveira MCV, da Silva KC, Gomes CR, Moreno AM, et al. Virulence profiles, phylogenetic background, and antibiotic resistance of Escherichia coli isolated from turkeys with airsacculitis. Sci World J. 2014;2014(1):289024. [DOI:10.1155/2014/289024] [PMID] [PMCID]
44. Ewers C, Li G, Wilking H, Kieβling S, Alt K, Antáo E-M, et al. Avian pathogenic, uropathogenic, and newborn meningitis-causing Escherichia coli: how closely related are they?. Int J Med Microbiol. 2007;297(3):163-76. [DOI:10.1016/j.ijmm.2007.01.003] [PMID]
45. Collingwood C, Kemmett K, Williams N, Wigley P. Is the concept of avian pathogenic Escherichia coli as a single pathotype fundamentally flawed?. Front Vet Sci. 2014;1:5. [DOI:10.3389/fvets.2014.00005] [PMID] [PMCID]
46. Sasoon A, Nikkhahi F, Javadi A, Sabzi S, Deilamani MO, Kiaheyrati N, et al. Biofilm Formation and Antibiotic Resistance Genes of Escherichia coli From Poultry Farms and Clinical Samples. Vet Med Sci. 2025;11(5):e70510. [DOI:10.1002/vms3.70510] [PMID] [PMCID]
47. Pavlickova S, Klancnik A, Dolezalova M, Mozina SS, Holko I. Antibiotic resistance, virulence factors and biofilm formation ability in Escherichia coli strains isolated from chicken meat and wildlife in the Czech Republic. J Environ Sci Health B. 2017;52(8):570-6. [DOI:10.1080/03601234.2017.1318637] [PMID]
48. Liu X, Yao H, Zhao X, Ge C. Biofilm formation and control of foodborne pathogenic bacteria. Molecules. 2023;28(6):2432. [DOI:10.3390/molecules28062432] [PMID] [PMCID]
49. Donlan RM, Costerton JW. Biofilms: survival mechanisms of clinically relevant microorganisms. Clin Microbiol Rev. 2002;15(2):167-93. [DOI:10.1128/CMR.15.2.167-193.2002] [PMID] [PMCID]
50. Marin C, Hernandiz A, Lainez M. Biofilm development capacity of Salmonella strains isolated in poultry risk factors and their resistance against disinfectants. Poult Sci. 2009;88(2):424-31. [DOI:10.3382/ps.2008-00241] [PMID]
51. Russell A. Glutaraldehyde: current status and uses. Infect Control Hosp Epidemiol. 1994;15(11):724-33. [DOI:10.1086/646845] [PMID]
52. Gorman S, Scott EM, Russell A. Antimicrobial activity, uses and mechanism of action of glutaraldehyde. J Appl Microbiol. 1980;48(2):161-90. [DOI:10.1111/j.1365-2672.1980.tb01217.x] [PMID]
53. Oosterik LH, Peeters L, Mutuku I, Goddeeris BM, Butaye P. Susceptibility of avian pathogenic Escherichia coli from laying hens in Belgium to antibiotics and disinfectants and integron prevalence. Avian Dis. 2014;58(2):271-8. [DOI:10.1637/10680-100113-RegR] [PMID]
54. Enany ME, Algammal AM, Nasef SA, Abo-Eillil SA, Bin-Jumah M, Taha AE, et al. The occurrence of the multidrug resistance (MDR) and the prevalence of virulence genes and QACs resistance genes in E. coli isolated from environmental and avian sources. AMB Express. 2019;9:1-9. [DOI:10.1186/s13568-019-0920-4] [PMID] [PMCID]
55. Bragg R, Jansen A, Coetzee M, van der Westhuizen W, Boucher C. Bacterial resistance to quaternary ammonium compounds (QAC) disinfectants. Infectious Diseases and Nanomedicine II: First International Conference (ICIDN-2012), Dec 15-18, 2012, Kathmandu, Nepal; 2014: Springer. [DOI:10.1007/978-81-322-1774-9_1] [PMID]
56. Kim K-H, Jahan SA, Lee J-T. Exposure to formaldehyde and its potential human health hazards. J Environ Sci Health C. 2011;29(4):277-99. [DOI:10.1080/10590501.2011.629972] [PMID]

Add your comments about this article : Your username or Email:
CAPTCHA

Send email to the article author


Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

© 2025 CC BY-NC 4.0 | Iranian Journal of Medical Microbiology

Designed & Developed by : Yektaweb | Publisher: Farname Inc