Exploring anti-quorum sensing potential and biofilm inhibitory effect of azithromycin against urinary Escherichia coli isolated from a teaching hospital in Alexandria
Morgan, H., Omar, H., Mohamed, N., Zakaria, A. (2024). Exploring anti-quorum sensing potential and biofilm inhibitory effect of azithromycin against urinary Escherichia coli isolated from a teaching hospital in Alexandria. EKB Journal Management System, (), 1-17. doi: 10.21608/japs.2024.271190.1017
Hadeer Aly Morgan; Hoda Mohamed Gamal Omar; Nelly Mostafa Mohamed; Azza Said Zakaria. "Exploring anti-quorum sensing potential and biofilm inhibitory effect of azithromycin against urinary Escherichia coli isolated from a teaching hospital in Alexandria". EKB Journal Management System, , , 2024, 1-17. doi: 10.21608/japs.2024.271190.1017
Morgan, H., Omar, H., Mohamed, N., Zakaria, A. (2024). 'Exploring anti-quorum sensing potential and biofilm inhibitory effect of azithromycin against urinary Escherichia coli isolated from a teaching hospital in Alexandria', EKB Journal Management System, (), pp. 1-17. doi: 10.21608/japs.2024.271190.1017
Morgan, H., Omar, H., Mohamed, N., Zakaria, A. Exploring anti-quorum sensing potential and biofilm inhibitory effect of azithromycin against urinary Escherichia coli isolated from a teaching hospital in Alexandria. EKB Journal Management System, 2024; (): 1-17. doi: 10.21608/japs.2024.271190.1017

Exploring anti-quorum sensing potential and biofilm inhibitory effect of azithromycin against urinary Escherichia coli isolated from a teaching hospital in Alexandria

Journal of Advanced Pharmaceutical Sciences
Articles in Press, Accepted Manuscript, Available Online from 31 March 2024  XML PDF (697.57 K)
Document Type: Original Article
DOI: 10.21608/japs.2024.271190.1017
View on SCiNiTO View on SCiNiTO
Authors
Hadeer Aly Morganorcid 1; Hoda Mohamed Gamal Omarorcid 2; Nelly Mostafa Mohamed email orcid 1; Azza Said Zakariaorcid 3
1Department of Microbiology and Immunology, Faculty of Pharmacy, Alexandria University, Alexandria, Egypt
2Department of Microbiology and Immunology Faculty of Pharmacy, Alexandria University, Alexandria, Egypt
3Department of Microbiology and Immunology Faculty of Pharmacy Alexandria University
Abstract
Urinary tract infections (UTIs) are the most prevalent infectious diseases in community and clinical settings. The global emergence of multidrug-resistant (MDR) E. coli causing these UTIs necessitates exploring novel approaches. The use of compounds that attenuate bacterial virulence by acting through inhibition of quorum sensing (QS) is gathering momentum nowadays. This research was undertaken to investigate the anti-QS prospects of azithromycin (AZ) against 67 E. coli isolated from urine cultures of patients admitted to Alexandria Main University Hospital. Antibiotic susceptibility testing showed that 94% of these isolates were MDR. Swimming, twitching, and biofilm formation abilities were detected in 60%, 52.5%, and 55.2%, of the isolates, respectively. At its sub-inhibitory concentration, AZ hindered swimming and twitching motilities in 75% and 74.3% of the tested isolates, respectively. It reduced the biofilm formation by percentages ranging from 27% to 73%. Azithromycin downregulated the differential gene expression of luxS, a QS-regulating gene, and the genes encoding motility, thus proving its anti-QS ability. Further, the combinatory activity of AZ with ceftriaxone, ciprofloxacin, colistin, doxycycline, and tigecycline generated 95% of successful combinations.
In conclusion, AZ effectively attenuated the QS in urinary E. coli isolates, leading to a regression in the production of QS-associated virulence factors and hindrance of biofilm formation, thus delivering an avirulent pathogen. In addition, the synergistic combinations succeeded in circumventing the antibiotic resistance of E. coli isolates. Hence, AZ could be regarded as an anti-QS agent that might aid the design of innovative treatment alternatives for the management of UTIs caused by MDR E. coli.

Highlights
  • Compounds attenuating bacterial virulence through inhibition of quorum sensing are gathering momentum nowadays.
  • Anti-quorum sensing potential of azithromycin against multidrug-resistant coli isolated from urine was investigated.
  • Azithromycin hindered the swimming and twitching motilities and reduced the biofilm formation in tested isolates.
  • It downregulated the expression of the quorum-sensing genes and genes encoding motility, thus proving its anti-quorum-sensing ability.
  • The combinatory activity of azithromycin with selected antibiotics generated successful combinations that could be beneficial for the treatment of urinary tract infections caused by E. coli.
Keywords
Azithromycin; E. coli; quorum sensing; differential gene expression; checkerboard assay
Supplementary Files
References
  1. Flores-Mireles A, Walker J, Caparon M, Hultgren S, Urinary tract infections: epidemiology, mechanisms of infection and treatment options. Nat Rev Microbiol, 2015, 13(5), 269-284.
  2. Silva A, Costa E, Freitas A, Almeida A, Revisiting the frequency and antimicrobial resistance patterns of bacteria implicated in community urinary tract infections. Antibiotics, 2022, 11(6), 768.
  3. Kaye KS, Gupta V, Mulgirigama A, Joshi AV, Scangarella-Oman NE, Yu K, Ye G, Mitrani-Gold FS, Antimicrobial resistance trends in urine Escherichia coli isolates from adult and adolescent females in the United States from 2011 to 2019: rising ESBL strains and impact on patient management. Clin Infect Dis, 2021, 73(11), 1992-1999.
  4. Escher N, Muhummed A, Hattendorf J, Vonaesch P, Zinsstag J, Systematic review and meta-analysis of integrated studies on antimicrobial resistance genes in Africa-A One Health perspective. Trop Med Int Health, 2021, 26(10), 1153-1163.
  5. Moghnieh RA, Kanafani ZA, Tabaja HZ, Sharara SL, Awad LS, Kanj SS, Epidemiology of common resistant bacterial pathogens in the countries of the Arab League. Lancet Infect Dis, 2018, 18(12), e379-e394.
  6. El-Kholy A, El-Mahallawy HA, Elsharnouby N, Abdel Aziz M, Helmy AM, Kotb R, Landscape of multidrug-resistant gram-negative infections in Egypt: survey and literature review. Infect Drug Resist, 2021, 14, 1905-1920.
  7. Alyousef AA, Mabood Husain F, Arshad M, Rizwan Ahamad S, Shavez Khan M, Abul Qais F, et al., Myrtus communis and its bioactive phytoconstituent, linalool, interferes with quorum sensing regulated virulence functions and biofilm of uropathogenic bacteria: In vitro and in silico insights. J King Saud Univ Sci, 2021, 33(7), 101588.
  8. Escobar-Muciño E, Arenas-Hernández MMP, Luna-Guevara ML, Mechanisms of inhibition of quorum sensing as an alternative for the control of coli and Salmonella. Microorganisms, 2022, 10(5), 884.
  9. Whelan S, Lucey B, Finn K, Uropathogenic Escherichia coli (UPEC)-associated urinary tract infections: The molecular basis for challenges to effective treatment. Microorganisms, 2023, 11(9), 2169.
  10. Aboushleib HM, Omar HM, Abozahra R, Elsheredy A, Baraka K, Correlation of quorum sensing and virulence factors in Pseudomonas aeruginosa isolates in Egypt. J Infect Dev Ctries, 2015, 9(10), 1091-1099.
  11. Ćirić A, Petrović J, Glamočlija J, Smiljković M, Nikolić M, Stojković D, et al., Natural products as biofilm formation antagonists and regulators of quorum sensing functions: A comprehensive review update and future trends. S Afr J Bot, 2019, 120, 65-80.
  12. Morgaan HA, Omar HMG, Zakaria AS, Mohamed NM, Repurposing carvacrol, cinnamaldehyde, and eugenol as potential anti-quorum sensing agents against uropathogenic Escherichia coli isolates in Alexandria, Egypt. BMC Microbiol, 2023, 23(1), 300.
  13. Elsheredy A, El-Soudany I, Elsherbini E, Metwally D, Ghazal A, Effect of azithromycin and phenylalanine-arginine beta-naphthylamide on quorum sensing and virulence factors in clinical isolates of Pseudomonas aeruginosa. Iran J Microbiol, 2021, 13(1), 37-49.
  14. Clinical and Laboratory Standards Institute (CLSI): Performance Standards for Antimicrobial Susceptibility Testing: Informational Supplement, M100. Clinical and Laboratory Standards Institute, 2018, Wayne P.
  15. (15). The European Committee on Antimicrobial Susceptibility Testing (EUCAST) (2018) Breakpoint tables for interpretation of MICs and zone diameters, version 8.0.
  16. Mohamed NM, Zakaria AS, Edward EA, Genomic characterization of international high-risk clone ST410 Escherichia coli co-harboring ESBL-encoding genes and blaNDM-5 on IncFIA/IncFIB/IncFII/IncQ1 multireplicon plasmid and carrying a chromosome-borne blaCMY-2 from Egypt. Antibiotics, 2022, 11(8), 1031.
  17. Lee JH, Kim YG, Lee J, Carvacrol‐rich oregano oil and thymol‐rich thyme red oil inhibit biofilm formation and the virulence of uropathogenic Escherichia coli. J Appl Microbiol, 2017, 123(6), 1420-1428.
  18. Seleem NM, Abd El Latif HK, Shaldam MA, El-Ganiny A, Drugs with new lease of life as quorum sensing inhibitors: for combating MDR Acinetobacter baumannii Eur J Clin Microbiol, 2020, 39(9), 1687-1702.
  19. Vijayakumar S, Rajenderan S, Laishram S, Anandan S, Balaji V, Biswas I, Biofilm formation and motility depend on the nature of the Acinetobacter baumannii clinical isolates. Front Public Health, 2016, 4(105), 1-9.
  20. Javed S, Mirani ZA, Pirzada ZA, Phylogenetic group B2 expressed significant biofilm formation among drug-resistant uropathogenic Escherichia coli. Libyan J Med, 2021, 16(1), 1-7.
  21. Baldiris-Avila R, Montes-Robledo A, Buelvas-MontesY, Phylogenetic classification, biofilm-forming capacity, virulence factors, and antimicrobial resistance in uropathogenic Escherichia coli (UPEC). Curr Microbiol, 2020, 77(11), 3361-3370.
  22. Pfaffl, M.W. A new mathematical model for relative quantification in real-time RT-PCR. Nucl. Acid Res., 2001, 29, 2002–2007
  23. (Wang Y-m, Kong L-c, Liu J, Ma H-x, Synergistic effect of eugenol with colistin against clinical isolated colistin-resistant Escherichia coli Antimicrob Resist Infect Control, 2018, 7(1), 1-9.
  24. Abozahra R, Gaballah A, Abdelhamid SM, Prevalence of the colistin resistance gene MCR-1 in colistin-resistant Klebsiella pneumoniae in Egypt. AIMS Microbiology, 9(2), 177-194 (2023).
  25. Zhu H, Chen Y , Hang Y, Luo H, Fang X, Xiao Y, et al., Impact of inappropriate empirical antibiotic treatment on clinical outcomes of urinary tract infections caused by Escherichia coli: a retrospective cohort study. J Glob Antimicrob Resist, 2021, 26, 148-153.
  26. Abdelaziz SM, Aboshanab KM, Yahia IS, Yassien MA, Hassouna NA, Correlation between the antibiotic resistance genes and susceptibility to antibiotics among the carbapenem-resistant Gram-negative pathogens. Antibiotics, 2021, 10(3), 255.
  27. Abdelkhalik AM, Agha MM, Zaki AM, Tahoun AT, Clinical and lab-assessed antibiotic resistance pattern of uropathogens among women with acute uncomplicated cystitis. Egypt J Hosp Med, 2018, 73(11), 7860-7868.
  28. Masoud SM, Abd El-Baky RM, Aly SA, Ibrahem RA, Co-existence of certain ESBLs, MBLs and plasmid mediated quinolone resistance genes among MDR coli isolated from different clinical specimens in Egypt. Antibiotics, 2021, 10(7), 835.
  29. El-Sokkary RH, Ramadan RA, El-Shabrawy M, El-Korashi LA, Elhawary A, Embarak S, et al., Community acquired pneumonia among adult patients at an Egyptian university hospital: bacterial etiology, susceptibility profile and evaluation of the response to initial empiric antibiotic therapy. Infec. Drug Resist, 2018, 11, 2141-2150.
  30. Masoud SM, Abd El-Baky RM, Aly SA, Ibrahem RA, Prevalence of multidrug-resistant Escherichia coli recovered from patients suffering from urinary tract infections. Nov Res Microbiol J, 2022, 6(2), 1543-1556.
  31. Iskandar K, Molinier L, Hallit S, Sartelli M, Catena F, Coccolini F, et al., Drivers of antibiotic resistance transmission in low- and middle-income countries from a “One Health” perspective—A review. Antibiotics, 2020, 9(7), 372.
  32. Abdel-Moaty MM, Mohamed WS, Abdel-All SM, El-Hendawy HH, Prevalence and molecular epidemiology of extended spectrum β-lactamase producing Escherichia coli from hospital and community settings in Egypt. J Appl Pharm Sci, 2016, 6(1), 42-47.
  33. Mahmoud AT, Salim MT, Ibrahem RA, Gabr A, Halby HM, Multiple drug resistance patterns in various phylogenetic groups of hospital-acquired uropathogenic coli isolated from cancer patients. Antibiotics, 2020, 9(3), 108.
  34. Mahgoub FM and El-Gamal S, Microbiological profile of urinary tract infections with special reference to antibiotic susceptibility pattern of Escherichia coli Int J Curr Microbiol App Sci, 2018, 7(2), 911-920.
  35. Fasugba O, Gardner A, Mitchell BG, Mnatzaganian G, Ciprofloxacin resistance in community- and hospital-acquired Escherichia coli urinary tract infections: a systematic review and meta-analysis of observational studies. BMC Infect Dis, 2015, 15(1), 545.
  36. Said A, El-Gamal MS, Abu-Elghait M, Salem SS, Isolation, identification and antibiotic susceptibility pattern of urinary tract infection bacterial isolates. Lett Appl NanoBioSci, 2021, 10, 2820-2830.
  37. Zakaria AS, Edward EA, Mohamed NM, Genomic insights into a colistin-resistant uropathogenic Escherichia coli strain of O23:H4-ST641 lineage harboring mcr-1.1 on a conjugative IncHI2 plasmid from Egypt. Microorganisms, 2021, 9(4), 799.
  38. Patel G, Bonomo R, “Stormy waters ahead”: global emergence of carbapenemases. Front Microbiol, 2013, 4, 1-17.
  39. Verma S, Patil SS, In vitro biofilm formation by uropathogenic bacteria and their antibiotic susceptibility pattern. J Krishna Inst Med Sci Univ, 2016, 5(3), 61-67.
  40. Mohamad EA, El Shalakany AH, Detection of biofilm formation in uropathogenic bacteria. Egypt J Med Microbiol, 2015, 24(1), 49-58.
  41. Abdaagire N, Chincholkar V, Kulkarni D, Nilekar S, Birajdar S, Biofilm production among uropathogenes and their antibiogram. Int J Pharm Bio Sci, 2014, 5(1), 261-266.
  42. Al-Tufaili AA, Manhal FS, Naeem AK, The correlation between some pathogenicity associated virulence factor and biofilm formation among uropathogenic Escherichia coli isolates in Al Najaf Al-Ashraf province. Al-Kufa Uni J for Biol, 2016, 8(2), 135-144.
  43. Zhang D, Gan R-Y, Zhang J-R, Farha AK, Li H-B, Zhu F, et al., Antivirulence properties and related mechanisms of spice essential oils: A comprehensive review. Compr Rev Food Sci Food Saf, 2020, 19(3), 1018-1055.
  44. Mihaylova M, Kostadinova S, Marhova M, Distribution of virulence determinants and biofilmforming among clinical urinary isolates. J Biosci Biotech, 2012, 45-51.
  45. Czaplewski L, Bax R, Clokie M, Dawson M, Fairhead H, Fischetti VA, et al., Alternatives to antibiotics—a pipeline portfolio review. Lancet Infect Dis, 2016, 16(2), 239-251.
  46. Swatton JE, Davenport PW, Maunders EA, Griffin JL, Lilley KS, Welch M, Impact of azithromycin on the quorum sensing-controlled proteome of Pseudomonas aeruginosa. PLOS ONE, 2016, 11(1), e0147698.
  47. Bala A, Kumar R, Harjai K, Inhibition of quorum sensing in Pseudomonas aeruginosa by azithromycin and its effectiveness in urinary tract infections. J Med Microbiol, 2011, 60(3), 300-306.
  48. Goyal S, Beniwal V, Kumar R, Kumar R, kumar A, Chhokar V, et al. Molecular typing of multidrug resistant uropathogenic Escherichia coli by restriction fragment length polymorphism. Eng Sci, 2022, 18, 204-216.
  49. Matsui H, Eguchi M, Ohsumi K, Nakamura A, Isshiki Y, Sekiya K, et al., Azithromycin inhibits the formation of flagellar filaments without suppressing flagellin synthesis in Salmonella enterica Serovar Typhimurium. Antimicrob Agents Chemother, 2005, 49(8), 3396-3403.
  50. Soto SM, Importance of biofilms in urinary tract infections: new therapeutic approaches. Adv Biol, 2014, 1-13.
  51. Henly EL, Norris K, Rawson K, Zoulias N, Jaques L, Chirila PG, et al., Impact of long-term quorum sensing inhibition on uropathogenic Escherichia coli. J Antimicrob Chemother, 2021, 76(4), 909-919.
  52. Ahsan S, Alam NE, Sirajee AS, Determination of the efficacy of azithromycin on biofilm-forming uropathogenic Escherichia coli isolated from urinary tract infection samples. Dhaka Univ J Biol Sci, 2021, 30(1), 1-11.
  53. Bala A, Kumar R, Harjai K, Inhibition of quorum sensing in Pseudomonas aeruginosa by azithromycin and its effectiveness in urinary tract infections. J Med Microbiol, 2011, 60(3), 300-306.
  54. Xu Z, Gao Y, He J, Xu W, Jiang M, Jin H, Effects of azithromycin on Pseudomonas aeruginosa isolates from catheter-associated urinary tract infection. Exp Ther Med, 2015, 9(2), 569-572.
  55. Sharifi A, Nayeri Fasaei B, Selected plant essential oils inhibit biofilm formation and luxS- and pfs-mediated quorum sensing by Escherichia coli O157:H7. Lett Appl Microbiol, 2002, 74(6), 916-923.
  56. Nalca Y, Jänsch L, Bredenbruch F, Geffers R, Buer J, Häussler S, Quorum-sensing antagonistic activities of azithromycin in Pseudomonas aeruginosa PAO1: A gobal Approach. Antimicrob Agents Chemother, 2006, 50(5), 1680-1688.
  57. Yang Y, Zhou M, Huayan H, Zhu J, Yao F, Zhang X, et al., Quorum-sensing gene luxS regulates flagella expression and Shiga-like toxin production in F18ab Escherichia coli. Can J Microbiol, 2014, 60, 1-7.
  58. Heidary M, Samangani A, Kargari A, Kiani Nejad A, Yashmi I, Motahar M, et al., Mechanism of action, resistance, synergism, and clinical implications of azithromycin. J Clin Lab Anal, 2022, 36(6), e24427.
  59. Gómara-Lomero M, López-Calleja A, Rezusta A, Aínsa J, Ramón-García S, Repurposing azithromycin in combination with last-line fosfomycin, colistin and tigecycline against multi-drug-resistant Klebsiella pneumoniae. bioRxiv, 2022.
  60. Singkham-in U, Muhummudaree N, Chatsuwan T, In vitro synergism of azithromycin combination with antibiotics against OXA-48-producing Klebsiella pneumoniae clinical isolates. Antibiotics, 2021, 10(12), 1-15.
  61. Li Y, Lin X, Yao X, Huang Y, Liu W, Ma T, et al., Synergistic antimicrobial activity of colistin in combination with rifampin and azithromycin against Escherichia coli producing mcr-1. Antimicrob Agents Chemother, 2018, 62(12).
  62. Okasha HAS, Meheissen MA, In vitro activity of colistin and vancomycin or azithromycin combinations on extensively drug-resistant Acinetobacter baumannii clinical isolates. J Adv Microbiol, 2017, 7(1), 71-81.
  63. Saini H, Chhibber S, Harjai K, Azithromycin and ciprofloxacin: A possible synergistic combination against Pseudomonas aeruginosa biofilm-associated urinary tract infections. Int J Antimicrob Agents, 2015, 45(4), 359-367.
  64. Bishr AS, Abdelaziz SM, Yahia IS, Yassien MA, Hassouna NA, Aboshanab KM, Association of macrolide resistance genotypes and synergistic antibiotic combinations for combating macrolide-resistant MRSA recovered from hospitalized patients. Biology, 2021, 10(7), 624.
  65. Dillon N, Holland M, Tsunemoto H, Hancock B, Cornax I, Pogliano J, et al., Surprising synergy of dual translation inhibition vs. Acinetobacter baumannii and other multidrug-resistant bacterial pathogens. eBioMedicine, 2019, 46, 193-201.
  66. Naziri Z, Kilegolan JA, Moezzi MS, Derakhshandeh A. Biofilm formation by uropathogenic Escherichia coli : A complicating factor for treatment and recurrence of urinary tract infections. J Hosp Infect. 2021;117:9-16.
  67. Vinothkannan R, Tamizh MM, Raj CD, Princy Sa. Fructose furoic acid ester: An effective quorum sensing inhibitor against uropathogenic Escherichia coli. Bioorg Chem. 2018;79:310-8.
  68. Xicohtencatl-Cortes J, Cruz-Córdova A, Cázares-Domínguez V, Escalona-Venegas G, Zavala-Vega S, Arellano-Galindo J, et al. Uropathogenic Escherichia coli strains harboring tosA gene were associated to high virulence genes and a multidrug-resistant profile. Microb Pathog. 2019;134:103593.
  69. Brennecke J, Kraut S, Zwadlo K, Gandi SK, Pritchard D, Templeton K, et al. High-yield extraction of Escherichia coli RNA from human whole blood. J Med Microbiol. 2017;66(3):301-11.
  70. Kim Y-G, Lee J-H, Park S, Kim S, Lee J. Inhibition of polymicrobial biofilm formation by saw palmetto oil, lauric acid and myristic acid. Microb Biotechnol. 2022;15(2):590-602.
Statistics
Article View: 33
PDF Download: 49