GENOTYPING OF QUORUM SENSING AND BIOFILM VIRULENCE FACTORS GENES IN LOCAL ISOLATES OF ACINETOBACTER BAUMANNII

Öz

Objective: The current investigation focused on Acinetobacter baumannii (A. baumanni), due to its growing significance as a hospital infection-causing pathogen and its resistance to several medications.

Material and Method: Sixty-five isolates of A. baumannii were isolated from wound samples of patients admitted to different hospitals in Baghdad between January and April of 2023. Two types of methods were used in the detection of biofilm formation: the first one was Congo red agar method and the second one was microtiter plate method. Genotypic detection of various virulence factors associated with A. baumannii was performed using monoplex, multiplex, and ERIC-PCR.

Result and Discussion: To use the PCR method to examine virulence genes like biofilm and quorum sensing (QS). The genes responsible for biofilm formation were identified by the PCR method, followed by ompA 63/65 (96.92%) and bap 48/65 (73.84%), whereas the genes responsible for chemical signals were found to be rhlI 43/65 (66.15%), LasI 58/65 (89.23%), LasR 56/65 (86.15%), and rhlR was 39/65 (60%) after quorum sensing (QS) system genes. ERIC-DNA fingerprinting's phylogenetic analysis illustrated the variety of all isolates by utilizing the Dice coefficient and the UPGM of phylogenetic analysis. Based on statistical analysis, the ERIC-PCR genotyping method correlation coefficient with the study virulence genes, antibiotic sensitivity test, and other variables of virulence was significant at p <0.05.

Anahtar Kelimeler

• Acinetobacter baumannii
• biofilm formation
• multidrug resistance
• virulence factors

YEREL ACİNETOBACTER BAUMANNİİ İZOLATLARINDA ÇOĞUNLUĞU ALGILAMA VE BİYOFİLM İLE İLİŞKİLİ VİRÜLANS GENLERİNİN GENOTİPLENDİRİLMESİ

Öz

Amaç: Mevcut araştırma, hastane enfeksiyonuna neden olan patojen olarak artan önemi ve çeşitli ilaçlara karşı direnci nedeniyle Acinetobacter baumannii (A. baumanni) üzerine odaklanmıştır. 

Gereç ve Yöntem: Ocak-Nisan 2023 tarihleri arasında Bağdat'taki farklı hastanelere başvuran hastaların yara örneklerinden altmış beş A. baumannii izole edilmiştir. Biyofilm oluşumunun tespitinde iki tür yöntem kullanılmıştır: Birincis Kongo kırmızısı agar yöntemi ve ikincisi mikrotitre plaka yöntemi. A. baumannii’ ye ait ait çeşitli virülans faktörlerinin genotipik tespiti monopleks, multipleks ve ERIC-PCR kullanılarak gerçekleştirilmiştir.

Sonuç ve Tartışma: PCR yöntemiyle biyofilm ve çoğunluğu algılama (quorum sensing; QS) ilişkili virülans genleri incelenmiştir. Biyofilm oluşumundan sorumlu genlerden ompA 63/65 (%96.92) ve bap 48/65 (%73.84) oranında tespit edilmiştir. Kimyasal sinyal mekanizmalarından sorumlu çoğunluğu algılama genlerinde ise rhlI 43/65 (%66.15), LasI 58/65 (%89.23), LasR 56/65 (%86.15) ve rhlR 39/65 (%60) bulunmuştur. ERIC-DNA parmak izi analizi, Dice katsayısı ve UPGMA filogenetik analiz yöntemiyle izolatların çeşitliliğini göstermiştir. İncelenen türler iki ayrı kümeye (A ve B) ayrılmıştır. İstatistiksel analizlere göre ERIC-PCR genotiplendirme yönteminin virülans genleri, antibiyotik duyarlılık testi ve diğer virülans değişkenleriyle korelasyon katsayısı anlamlı bulunmuştur (p < 0.05).

Anahtar Kelimeler

• Acinetobacter baumannii
• biyofim oluşumu
• çoklu ilaç direnci
• virülans faktörleri

Kaynakça

1. 1. Murray, L.M., Hayes, A., Snape, J., et al. (2024). Co-selection for antibiotic resistance by environmental contaminants. Npj Antimicrobials and Resistance, 2(1), 9. [CrossRef]
2. 2. Scoffone, V.C., Trespidi, G., Barbieri, G., et al. (2025). The evolution of antimicrobial resistance in Acinetobacter baumannii and new strategies to fight it. Antibiotics, 14(1), 85. [CrossRef]
3. 3. Karampatakis, T., Tsergouli, K., Behzadi, P. (2024). Pan-genome plasticity and virulence factors: A natural treasure trove for Acinetobacter baumannii. Antibiotics, 13(3), 257. [CrossRef]
4. 4. Attili, A.R., Nocera, F.P., Sisto, M., et al. (2024). Evidence and antibiotic resistance profiles of clinical Acinetobacter calcoaceticus-Acinetobacter baumannii (ACB) and non-ACB complex members in companion animals: A 2020-2022 retrospective study. Comparative Immunology, Microbiology and Infectious Diseases, 109, 102185. [CrossRef]
5. 5. Zhang, L., Tian, X., Sun, L., et al. (2024). Bacterial efflux pump inhibitors reduce antibiotic resistance. Pharmaceutics, 16(2), 170. [CrossRef]
6. 6. Anwar, M.S., Biswas, P.P., Sen, A. (2024). A literature review on Acinetobacter baumannii and the challenges: Insights into various clinical and pathophysiological conditions with reference to biofilm formation. Student’s Journal of Health Research Africa, 5(9), 1-14.[CrossRef]
7. 7. Zhang, R., Li, D., Fang, H., et al. (2025). Iron-dependent mechanisms in Acinetobacter baumannii: pathogenicity and resistance. JAC-Antimicrobial Resistance, 7(2). [CrossRef]
8. 8. Baleivanualala, S.C., Matanitobua, S., Soqo, V., et al. (2024). Molecular and clinical epidemiology of carbapenem resistant Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacterales in Fiji: A multicentre prospective observational study. The Lancet Regional Health - Western Pacific, 47, 101095. [CrossRef]

9. 9. Bansal, K., Saroha, T., Patil, P.P., et al. (2024). Evolutionary trends of carbapenem-resistant and susceptible Acinetobacter baumannii isolates in a major tertiary care setting from North India. Infection, Genetics and Evolution, 117, 105542. [CrossRef]
10. 10. Iovleva, A., Fowler, V.G., Doi, Y. (2024). Treatment approaches for carbapenem-resistant Acinetobacter baumannii infections. Drugs, 85(1), 21-40. [CrossRef]
11. 11. Marotta, J., Zhao, A., Rather, P.N., et al. (2025). The BfmRS stress response protects Acinetobacter baumannii against defects in outer membrane lipoprotein biogenesis. Journal of Bacteriology, 207(1). [CrossRef]
12. 12. Rajangam, S.L.,Narasimhan, M.K. (2024). Current treatment strategies for targeting virulence factors and biofilm formation in Acinetobacter baumannii. Future Microbiology, 19(10), 941-961. [CrossRef]
13. 13. Islam, M.M., Jung, D.E., Shin, W.S., et al. (2024). Colistin resistance mechanism and management strategies of colistin-resistant Acinetobacter baumannii infections. Pathogens, 13(12), 1049. [CrossRef]
14. 14. Wang, Y., Ling, N., Wang, Y., et al. (2024). Effect of ferric ions on cronobacter sakazakii growth, biofilm formation, and swarming motility. International Journal of Food Microbiology, 408, 110418. [CrossRef]
15. 15. Blanco-Romero, E., Garrido-Sanz, D., Durán, D., et al. (2024). Role of extracellular matrix components in biofilm formation and adaptation of Pseudomonas ogarae F113 to the rhizosphere environment. Frontiers in Microbiology, 15, 1341728. [CrossRef]
16. 16. Khoso, A., Hussain, A., Rehman, M., et al. (2024). Molecular assessments of antimicrobial protein enterocins and quorum sensing genes and their role in virulence of the genus enterococcus. Probiotics and Antimicrobial Proteins, 17(5), 3144-3155. [CrossRef]
17. 17. Juszczuk-Kubiak, E. (2024). Molecular aspects of the functioning of pathogenic bacteria biofilm based on quorum sensing (qs) signal-response system and innovative non-antibiotic strategies for their elimination. International Journal of Molecular Sciences, 25(5), 2655. [CrossRef]
18. 18. Jeong, G.J., Khan, F., Tabassum, N., Kim, Y.M. (2023). Motility of Acinetobacter baumannii: Regulatory systems and controlling strategies. Applied Microbiology and Biotechnology, 108(1), 3. [CrossRef]
19. 19. Mohammed, M.K., Ali, A.M., Albaayit, S.F.A. (2025). Detection of antimicrobial activity of Aspergillus terreus against clinical isolates of Serratia marcescens. The Eurasia Proceedings of Science Technology Engineering and Mathematics, 34, 93-101.‏ [CrossRef]
20. 20. Page, R., Peti, W.(2016). Toxin-antitoxin systems in bacterial growth arrest and persistence. Nature Chemical Biology, 12(4), 208-214. [CrossRef]
21. 21. Ghellai, L., Hassaine, H., Klouche, et al. (2014). Detection of biofilm formation of a collection of fifty strains of Staphylococcus aureus isolated in Algeria at the University Hospital of Tlemcen. Journal of Bacteriology Research, 6(1), 1-6.‏ [CrossRef]
22. 22. Abdel, H., Rania M., Nevine N.K., et al. (2018). Detection of biofilm producing staphylococci among different clinical isolates and its relation to methicillin susceptibility. Open Access Macedonian Journal of Medical Sciences, 6(8), 1335-1341. [CrossRef]
23. 23. Ali, A.M., Al-Kenanei, K.A., Hussein, S.N. et al. (2020). Molecular study of some virulence genes of Pseudomonas aeruginosa isolated from different infections in hospitals of Baghdad. Reviews in Medical Microbiology, 31(1), 26-41. [CrossRef]
24. 24. Hemmati, J., Nazari, M., Abolhasani, F.S., et al. (2024). In vitro investigation of relationship between quorum-sensing system genes, biofilm forming ability, and drug resistance in clinical isolates of Pseudomonas aeruginosa. BMC Microbiology, 24(1), 99. [CrossRef]
25. 25. Badmasti, F., Siadat, S.D., Bouzari, S., et al. (2015). Molecular detection of genes related to biofilm formation in multidrug-resistant Acinetobacter baumannii isolated from clinical settings. Journal of Medical Microbiology, 64, 559-564. [CrossRef]
26. 26. Ali, A.M., Al-kakei, S.N. (2019). Virulence factors genotyping of Klebsiella pneumoniae clinical isolates from Baghdad. Reviews in Medical Microbiology, 30(1), 36-46. [CrossRef]
27. 27. Kariuki, S. (2024). Global burden of antimicrobial resistance and forecasts to 2050. The Lancet, 404(10459), 1172-1173. [CrossRef]
28. 28. Bdaiwi, Q.O., Kareem, A.A., Hussein, S.N., et al. (2019). Prevalence of β-lactam resistant Klebsiella pneumoniae isolates among clinical specimensin Baghdad hospitals. Indian Journal of Public Health Research & Development, 10(11), 4935. [CrossRef]
29. 29. Huang, E., Thompson, R.N., Moon, S.H., et al. (2024). Treatment-emergent cefiderocol resistance in carbapenem-resistant Acinetobacter baumannii is associated with insertion sequence IS Aba36 in the siderophore receptor pirA. Antimicrobial Agents and Chemotherapy, 68(7), e0029024. [CrossRef]
30. 30. Alanazi, F.A., Almaary, K.S., Dawoud, T.M., et al. (2025). Molecular characterization of putative antibiotic resistance determinant and virulence factors genes of Acinetobacter baumannii strains isolated from intensive care unit patients in Riyadh, Saudi Arabia. Journal of Infection and Public Health, 18(4), 102695. [CrossRef]
31. 31. ‏Okesanya, O.J., Ahmed, M.M., Ogaya, J.B., et al. (2025). Reinvigorating AMR resilience: Leveraging CRISPR–Cas technology potentials to combat the 2024 WHO bacterial priority pathogens for enhanced global health security-a systematic review. Tropical Medicine and Health, 53(1), 43. [CrossRef]
32. 32. Zhang, R., Li, D., Fang, H., et al. (2025). Iron-dependent mechanisms in Acinetobacter baumannii: Pathogenicity and resistance. JAC-Antimicrobial Resistance, 7(2), dlaf039. [CrossRef]
33. 33. Liew, K.J., Shahar, S., Shamsir, M.S., et al. (2024). Integrating multi-platform assembly to recover MAGs from hot spring biofilms: insights into microbial diversity, biofilm formation, and carbohydrate degradation. Environmental Microbiome, 19(1), 29. [CrossRef]
34. 34. Sung, J.Y. (2018). Molecular characterization and antimicrobial susceptibility of biofilm-forming Acinetobacter baumannii clinical isolates from Daejeon, Korea. Korean Journal of Clinical Laboratory Science, 50(2), 100-109. [CrossRef]
35. 35. Chakraverty, R., Kundu, A.K. (2024). Hospital-acquired Infections in Intensive Care Unit and Their Management, Springer, Singapore, 1-12. [CrossRef]
36. 36. Lucidi, M., Visaggio, D., Migliaccio, A., et al. (2023). Pathogenicity and virulence of Acinetobacter baumannii: Factors contributing to the fitness in healthcare settings and the infected host. Virulence, 15(1), 2289769. [CrossRef]
37. 37. Marino, A., Augello, E., Stracquadanio, S., et al. (2024). Unveiling the secrets of Acinetobacter baumannii: resistance, current treatments, and future innovations. International Journal of Molecular Sciences, 25(13), 6814. [CrossRef]
38. 38. Bagińska, N., Grygiel, I., Orwat, F., et al. (2024). Stability study in selected conditions and biofilm-reducing activity of phages active against drug-resistant Acinetobacter baumannii. Scientific Reports, 14(1), 4285. [CrossRef]
39. 39. Yamada, N., Kamoshida, G., Shiraishi, T., et al. (2024). PmrAB, the two-component system of Acinetobacter baumannii, controls the phosphoethanolamine modification of lipooligosaccharide in response to metal ions. Journal of Bacteriology, 206(5), e0043523. [CrossRef]
40. 40. Mellace, M., Ceniti, C., Paonessa, M., et al., (2024). Multidrug-resistant Acinetobacter baumannii: An underestimated pathogen in veterinary medicine in Italy. German Journal of Veterinary Research, 4(2), 112-126. [CrossRef]
41. 41. Scribano, D., Cheri, E., Pompilio, A., et al. (2024). Acinetobacter baumannii OmpA-like porins: Functional characterization of bacterial physiology, antibiotic-resistance, and virulence. Communications Biology, 7(1), 948. [CrossRef]
42. 42. Kim, K., Islam, Md.M., Bang, S., et al. (2024). H-NS is a transcriptional repressor of the CRISPR-Cas system in Acinetobacter baumannii ATCC 19606. Journal of Microbiology, 62(11), 999-1012. [CrossRef]
43. 43. Sindhu, S. (2018). Clinical Significance, Antibiotic Resistance and Biofilm Formation of Acinetobacter baumannii: Review. Clinical Microbiology: Open Access, 7(4), 1000315. [CrossRef]
44. 44. Singh, S., Singh, S., Trivedi, M., et al. (2024). An insight into MDR Acinetobacter baumannii infection and its pathogenesis: Potential therapeutic targets and challenges. Microbial Pathogenesis, 192, 106674. [CrossRef]
45. 45. Khalif, O.I., Alkalifawi, E.J. (2025). Green-synthesized zinc oxide nanoparticles for Acinetobacter baumannii control: A review of plant-based approaches. Cureus, 17(5), e83506. [CrossRef]
46. 46. Subbarayudu, S., Snega Priya, P., Rajagopal, R., et al. (2024). Impact of acidic and alkaline conditions on Staphylococcus aureus and Acinetobacter baumannii interactions and their biofilms. Archives of Microbiology, 206(11), 426. [CrossRef]
47. 47. Al-Draghi, W.A., Wahab, S. A., Hamad, S.T. (2022). Effects of meropenem at sub-inhibitory doses on the expression of biofilm-associated protein by Acinetobacter baumannii. Internationa Journal of Drug Delivery Technology, 12(02), 841-845. [CrossRef]