Diversity of biofilm-specific antimicrobial resistance genes in Pseudomonas aeruginosa recovered from various clinical isolates
,
Abstract
Background and Objectives: The resistance of Pseudomonas aeruginosa to antibiotics offers a significant challenge in the treatment of patients. This study aimed to investigate the antimicrobial resistance profile, biofilm-specific antimicrobial resistance genes, and genetic diversity of P. aeruginosa recovered from clinical samples.
Materials and Methods: Totally 47 non-duplicate isolates of P. aeruginosa were recovered from various clinical samples. toxA, algD, ndvB, and tssC1 genes were detected in biofilm-producing isolates. The DNA sequences of the toxA and tssC1 genes were analyzed, by creating phylogenetic trees.
Results: The findings revealed that 30 (63.8%) of the isolates tested positive for Extended spectrum β-lactamase (ESBL), whereas 31 (65.9%) tested positive for Metallo-β-lactamase (MBL) and all of the isolates presented the toxA genes, and 19.1%,17%, 6.3% presented by algD, ndvB and tssC1 genes. Besides, the phylogenetic trees of the toxA and tssC1 gene isolates suggested a genotype that was closely aligned with others. Gene sequencing similarity revealed 99% identity with other isolates deposited in GenBank.
Conclusion: The occurrence of toxA was most prevalent. One isolate was recorded as a novel isolate in the global gene bank as a locally isolated strain from the city of Erbil that has never been identified in global isolates due to genetic variation.
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6. Soto SM. Role of efflux pumps in the antibiotic resistance of bacteria embedded in a biofilm. Virulence 2013; 4: 223-229.
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8. Fazeli N, Momtaz H. Virulence gene profiles of multidrug-resistant Pseudomonas aeruginosa isolated from Iranian hospital infections. Iran Red Crescent Med J 2014; 16(10): e15722.
9. Arslan E, Coşkun MK, Çobanoğlu Ş, Aslan MH, Yazıcı A. Effects of four antibiotics on Pseudomonas aeruginosa motility, biofilm formation, and biofilm-specific antibiotic resistance genes expression. Diagn Microbiol Infect Dis 2023; 106: 115931.
10. Pang Z, Raudonis R, Glick BR, Lin T-J, Cheng Z. Antibiotic resistance in Pseudomonas aeruginosa: mechanisms and alternative therapeutic strategies. Biotechnol Adv 2019; 37: 177-192.
11. Humphries RM, Abbott AN, Hindler JA. Understanding and addressing CLSI breakpoint revisions: a primer for clinical laboratories. J Clin Microbiol 2019; 57(6): e00203-19.
12. Vahdani M, Azimi L, Asghari B, Bazmi F, Lari AR. Phenotypic screening of extended-spectrum ß-lactamase and metallo-ß-lactamase in multidrug-resistant Pseudomonas aeruginosa from infected burns. Ann Burns Fire Disasters 2012; 25: 78-81.
13. Montes-Robledo A, Buelvas-Montes Y, Baldiris-Avila R. Description of extraintestinal pathogenic Escherichia coli based on phylogenetic grouping, virulence factors, and antimicrobial susceptibility. Iran J Microbiol 2023; 15: 503-512.
14. Dumaru R, Baral R, Shrestha LB. Study of biofilm formation and antibiotic resistance pattern of gram-negative Bacilli among the clinical isolates at BPKIHS, Dharan. BMC Res Notes 2019; 12: 38.
15. Khidhr Rahman J, Ahmed Ahmed A, Ganjo AR, Salih Mohamad T. Assessment of toxicity, anti-quorum sensing and anti-biofilm production effects of Hypericum triquetrifolium Turra extract on multi-drug resistant Acinetobacter baumannii. J King Saud Univ Sci 2023; 35: 102714.
16. Saffari M, Karami S, Firoozeh F, Sehat M. Evaluation of biofilm-specific antimicrobial resistance genes in Pseudomonas aeruginosa isolates in Farabi Hospital. J Med Microbiol 2017; 66: 905-909.
17. Jaafar ZM, Dhahi MA, Abd AKH, Jaafar SM. Molecular identification and antibiotics resistance genes profile of Pseudomonas aeruginosa isolated from Iraqi patients. Afr J Microbiol Res 2014; 8: 2183-2192.
18. Telling K, Laht M, Brauer A, Remm M, Kisand V, Maimets M, et al. Multidrug resistant Pseudomonas aeruginosa in Estonian hospitals. BMC Infect Dis 2018; 18: 513.
19. Nassar O, Desouky SE, El-Sherbiny GM, Abu-Elghait M. Correlation between phenotypic virulence traits and antibiotic resistance in Pseudomonas aeruginosa clinical isolates. Microb Pathog 2022; 162: 105339.
20. Izadi Pour Jahromi S, Mardaneh J, Sharifi A, Pezeshkpour V, Behzad-Behbahani A, Seyyedi N, et al. Occurrence of a multidrug resistant Pseudomonas aeruginosa strains in hospitalized patients in southwest of Iran: Characterization of resistance trends and virulence determinants. Jundishapur J Microbiol 2018; 11(4): e57341.
21. Alvarez-Ortega C, Wiegand I, Olivares J, Hancock RE, Martínez JL. The intrinsic resistome of Pseudomonas aeruginosa to β-lactams. Virulence 2011; 2: 144-146.
22. Pachori P, Gothalwal R, Gandhi P. Emergence of antibiotic resistance Pseudomonas aeruginosa in intensive care unit; a critical review. Genes Dis 2019; 6: 109-119.
23. Terreni M, Taccani M, Pregnolato M. New antibiotics for multidrug-resistant bacterial strains: Latest research developments and future perspectives. Molecules 2021; 26: 2671.
24. El-sayed H, Fahmy Y. Correlation between biofilm formation and multidrug resistance in clinical isolates of Pseudomonas aeruginosa. Microb Infect Dis 2021; 2: 541-549.
25. Kamali E, Jamali A, Ardebili A, Ezadi F, Mohebbi A. Evaluation of antimicrobial resistance, biofilm forming potential, and the presence of biofilm-related genes among clinical isolates of Pseudomonas aeruginosa. BMC Res Notes 2020; 13: 27.
26. Ali FA. Association between biofilm formation gene bla exoU and metallo and extend spectrum beta-lactamase production of multidrug resistance Pseudomonas aeruginosa in clinical samples. Comb Chem High Throughput Screen 2022; 25: 1207-1218.
27. Ghadiri H, Vaez H, Razavi-Azarkhiavi K, Rezaee R, Haji-Noormohammadi M, Rahimi AA, et al. Prevalence and antibiotic susceptibility patterns of Extended-Spectrum ß-Lactamase and Metallo-ß-Lactamase–producing uropathogenic Escherichia coli isolates. Lab Med 2014; 45: 291-296.
28. Bahador N, Shoja S, Faridi F, Dozandeh-Mobarrez B, Qeshmi FI, Javadpour S, et al. Molecular detection of virulence factors and biofilm formation in Pseudomonas aeruginosa obtained from different clinical specimens in Bandar Abbas. Iran J Microbiol 2019; 11: 25-30.
29. Khattab MA, Nour MS, ElSheshtawy NM. Genetic identification of Pseudomonas aeruginosa virulence genes among different isolates. J Microb Biochem Technol 2015; 7: 274-277.
30. Hassuna NA, Mandour SA, Mohamed ES. Virulence constitution of multi-drug-resistant Pseudomonas aeruginosa in upper Egypt. Infect Drug Resist 2020; 13: 587-595.
31. Karna SLR, Nguyen JQ, Evani SJ, Qian L-W, Chen P, Abercrombie JJ, et al. T3SS and alginate biosynthesis of Pseudomonas aeruginosa impair healing of infected rabbit wounds. Microb Pathog 2020; 147: 104254.
32. Whitney JC, Whitfield GB, Marmont LS, Yip P, Neculai AM, Lobsanov YD, et al. Dimeric c-di-GMP is required for post-translational regulation of alginate production in Pseudomonas aeruginosa. J Biol Chem 2015; 290: 12451-12462.
33. Pottier M, Castagnet S, Gravey F, Leduc G, Sévin C, Petry S, et al. Antimicrobial resistance and genetic diversity of Pseudomonas aeruginosa strains isolated from equine and other veterinary samples. Pathogens 2022; 12: 64.
34. Elmouaden C, Laglaoui A, Ennanei L, Bakkali M, Abid M. Virulence genes and antibiotic resistance of Pseudomonas aeruginosa isolated from patients in the Northwestern of Morocco. J Infect Dev Ctries 2019; 13: 892-898.
35. Zhang L, Hinz AJ, Nadeau J-P, Mah T-F. Pseudomonas aeruginosa tssC1 links type VI secretion and biofilm-specific antibiotic resistance. J Bacteriol 2011; 193: 5510-5513.
36. Hall CW, Hinz AJ, Gagnon LB, Zhang L, Nadeau J-P, Copeland S, et al. Pseudomonas aeruginosa biofilm antibiotic resistance gene ndvB expression requires the RpoS stationary-phase sigma factor. Appl Environ Microbiol 2018; 84(7): e02762-17.
2. Krychowiak-Maśnicka M, Krauze-Baranowska M, Godlewska S, Kaczyński Z, Bielicka-Giełdoń A, Grzegorczyk N, et al. Potential of silver nanoparticles in overcoming the intrinsic resistance of Pseudomonas aeruginosa to secondary metabolites from carnivorous plants. Int J Mol Sci 2021; 22: 4849.
3. Meletis G, Exindari M, Vavatsi N, Sofianou D, Diza E. Mechanisms responsible for the emergence of carbapenem resistance in Pseudomonas aeruginosa. Hippokratia 2012; 16: 303-307.
4. Litwin A, Rojek S, Gozdzik W, Duszynska W. Pseudomonas aeruginosa device associated–healthcare associated infections and its multidrug resistance at intensive care unit of University Hospital: polish, 8.5-year, prospective, single-centre study. BMC Infect Dis 2021; 21: 180.
5. Høiby N, Bjarnsholt T, Givskov M, Molin S, Ciofu O. Antibiotic resistance of bacterial biofilms. Int J Antimicrob Agents 2010; 35: 322-332.
6. Soto SM. Role of efflux pumps in the antibiotic resistance of bacteria embedded in a biofilm. Virulence 2013; 4: 223-229.
7. Ratajczak M, Kamińska D, Nowak-Malczewska DM, Schneider A, Dlugaszewska J. Relationship between antibiotic resistance, biofilm formation, genes coding virulence factors and source of origin of Pseudomonas aeruginosa in clinical strains. Ann Agric Environ Med 2021; 28: 306-313.
8. Fazeli N, Momtaz H. Virulence gene profiles of multidrug-resistant Pseudomonas aeruginosa isolated from Iranian hospital infections. Iran Red Crescent Med J 2014; 16(10): e15722.
9. Arslan E, Coşkun MK, Çobanoğlu Ş, Aslan MH, Yazıcı A. Effects of four antibiotics on Pseudomonas aeruginosa motility, biofilm formation, and biofilm-specific antibiotic resistance genes expression. Diagn Microbiol Infect Dis 2023; 106: 115931.
10. Pang Z, Raudonis R, Glick BR, Lin T-J, Cheng Z. Antibiotic resistance in Pseudomonas aeruginosa: mechanisms and alternative therapeutic strategies. Biotechnol Adv 2019; 37: 177-192.
11. Humphries RM, Abbott AN, Hindler JA. Understanding and addressing CLSI breakpoint revisions: a primer for clinical laboratories. J Clin Microbiol 2019; 57(6): e00203-19.
12. Vahdani M, Azimi L, Asghari B, Bazmi F, Lari AR. Phenotypic screening of extended-spectrum ß-lactamase and metallo-ß-lactamase in multidrug-resistant Pseudomonas aeruginosa from infected burns. Ann Burns Fire Disasters 2012; 25: 78-81.
13. Montes-Robledo A, Buelvas-Montes Y, Baldiris-Avila R. Description of extraintestinal pathogenic Escherichia coli based on phylogenetic grouping, virulence factors, and antimicrobial susceptibility. Iran J Microbiol 2023; 15: 503-512.
14. Dumaru R, Baral R, Shrestha LB. Study of biofilm formation and antibiotic resistance pattern of gram-negative Bacilli among the clinical isolates at BPKIHS, Dharan. BMC Res Notes 2019; 12: 38.
15. Khidhr Rahman J, Ahmed Ahmed A, Ganjo AR, Salih Mohamad T. Assessment of toxicity, anti-quorum sensing and anti-biofilm production effects of Hypericum triquetrifolium Turra extract on multi-drug resistant Acinetobacter baumannii. J King Saud Univ Sci 2023; 35: 102714.
16. Saffari M, Karami S, Firoozeh F, Sehat M. Evaluation of biofilm-specific antimicrobial resistance genes in Pseudomonas aeruginosa isolates in Farabi Hospital. J Med Microbiol 2017; 66: 905-909.
17. Jaafar ZM, Dhahi MA, Abd AKH, Jaafar SM. Molecular identification and antibiotics resistance genes profile of Pseudomonas aeruginosa isolated from Iraqi patients. Afr J Microbiol Res 2014; 8: 2183-2192.
18. Telling K, Laht M, Brauer A, Remm M, Kisand V, Maimets M, et al. Multidrug resistant Pseudomonas aeruginosa in Estonian hospitals. BMC Infect Dis 2018; 18: 513.
19. Nassar O, Desouky SE, El-Sherbiny GM, Abu-Elghait M. Correlation between phenotypic virulence traits and antibiotic resistance in Pseudomonas aeruginosa clinical isolates. Microb Pathog 2022; 162: 105339.
20. Izadi Pour Jahromi S, Mardaneh J, Sharifi A, Pezeshkpour V, Behzad-Behbahani A, Seyyedi N, et al. Occurrence of a multidrug resistant Pseudomonas aeruginosa strains in hospitalized patients in southwest of Iran: Characterization of resistance trends and virulence determinants. Jundishapur J Microbiol 2018; 11(4): e57341.
21. Alvarez-Ortega C, Wiegand I, Olivares J, Hancock RE, Martínez JL. The intrinsic resistome of Pseudomonas aeruginosa to β-lactams. Virulence 2011; 2: 144-146.
22. Pachori P, Gothalwal R, Gandhi P. Emergence of antibiotic resistance Pseudomonas aeruginosa in intensive care unit; a critical review. Genes Dis 2019; 6: 109-119.
23. Terreni M, Taccani M, Pregnolato M. New antibiotics for multidrug-resistant bacterial strains: Latest research developments and future perspectives. Molecules 2021; 26: 2671.
24. El-sayed H, Fahmy Y. Correlation between biofilm formation and multidrug resistance in clinical isolates of Pseudomonas aeruginosa. Microb Infect Dis 2021; 2: 541-549.
25. Kamali E, Jamali A, Ardebili A, Ezadi F, Mohebbi A. Evaluation of antimicrobial resistance, biofilm forming potential, and the presence of biofilm-related genes among clinical isolates of Pseudomonas aeruginosa. BMC Res Notes 2020; 13: 27.
26. Ali FA. Association between biofilm formation gene bla exoU and metallo and extend spectrum beta-lactamase production of multidrug resistance Pseudomonas aeruginosa in clinical samples. Comb Chem High Throughput Screen 2022; 25: 1207-1218.
27. Ghadiri H, Vaez H, Razavi-Azarkhiavi K, Rezaee R, Haji-Noormohammadi M, Rahimi AA, et al. Prevalence and antibiotic susceptibility patterns of Extended-Spectrum ß-Lactamase and Metallo-ß-Lactamase–producing uropathogenic Escherichia coli isolates. Lab Med 2014; 45: 291-296.
28. Bahador N, Shoja S, Faridi F, Dozandeh-Mobarrez B, Qeshmi FI, Javadpour S, et al. Molecular detection of virulence factors and biofilm formation in Pseudomonas aeruginosa obtained from different clinical specimens in Bandar Abbas. Iran J Microbiol 2019; 11: 25-30.
29. Khattab MA, Nour MS, ElSheshtawy NM. Genetic identification of Pseudomonas aeruginosa virulence genes among different isolates. J Microb Biochem Technol 2015; 7: 274-277.
30. Hassuna NA, Mandour SA, Mohamed ES. Virulence constitution of multi-drug-resistant Pseudomonas aeruginosa in upper Egypt. Infect Drug Resist 2020; 13: 587-595.
31. Karna SLR, Nguyen JQ, Evani SJ, Qian L-W, Chen P, Abercrombie JJ, et al. T3SS and alginate biosynthesis of Pseudomonas aeruginosa impair healing of infected rabbit wounds. Microb Pathog 2020; 147: 104254.
32. Whitney JC, Whitfield GB, Marmont LS, Yip P, Neculai AM, Lobsanov YD, et al. Dimeric c-di-GMP is required for post-translational regulation of alginate production in Pseudomonas aeruginosa. J Biol Chem 2015; 290: 12451-12462.
33. Pottier M, Castagnet S, Gravey F, Leduc G, Sévin C, Petry S, et al. Antimicrobial resistance and genetic diversity of Pseudomonas aeruginosa strains isolated from equine and other veterinary samples. Pathogens 2022; 12: 64.
34. Elmouaden C, Laglaoui A, Ennanei L, Bakkali M, Abid M. Virulence genes and antibiotic resistance of Pseudomonas aeruginosa isolated from patients in the Northwestern of Morocco. J Infect Dev Ctries 2019; 13: 892-898.
35. Zhang L, Hinz AJ, Nadeau J-P, Mah T-F. Pseudomonas aeruginosa tssC1 links type VI secretion and biofilm-specific antibiotic resistance. J Bacteriol 2011; 193: 5510-5513.
36. Hall CW, Hinz AJ, Gagnon LB, Zhang L, Nadeau J-P, Copeland S, et al. Pseudomonas aeruginosa biofilm antibiotic resistance gene ndvB expression requires the RpoS stationary-phase sigma factor. Appl Environ Microbiol 2018; 84(7): e02762-17.
| Files | ||
| Issue | Vol 15 No 6 (2023) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v15i6.14134 | |
| Keywords | ||
| Biofilm; Drug resistance; Genetic variation; Pseudomonas aeruginosa; Virulence factors | ||
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How to Cite
1.
Ganjo A, Ali F, Aka S, Hussen B, Smail S. Diversity of biofilm-specific antimicrobial resistance genes in Pseudomonas aeruginosa recovered from various clinical isolates. Iran J Microbiol. 2023;15(6):742-749.
