Characterization of Biofilm production and quorum sensing phenomenon among antibiotic resistant Acinetobacter baumannii isolated from wound infections in Iraq

Rawa Abdul Redha Aziz, Sawsan Sajid Al-Jubori

Abstract

Objective(s): Acinetobacter baumannii has emerged as a significant hospital pathogen, quickly becoming resistant to wide range of antimicrobials. It has gained notoriety as a cause of debilitating soft tissue infections in soldiers returning from Iraq and Afghanistan.This study focused on the characterization of biofilm production in Iraqi A. baumannii and its regulation by quorum sensing. Material and Methods: twenty four isolates were collected from Baghdad and Al-Najaf hospitals and re-diagnosed by compact Vitek 2 and genetically using housekeeping gene (recA). Phenotypic detection of biofilm formation was screened by microtiter dish assay, twitching motility assay, and SEM. Quorum sensing phenomenon (QS) was detected using conventional PCR. Results: 100% of isolates formed either strong or weak biofilm by microtiter dish assay. Twitching motility test revealed that 79.2% of isolates were motile. SEM analysis showed 79.2% of isolates made different stages of biofilm started with adhesion step and ending with a mushroom like architecture as highly magnification images showed on glass cover slips. PCR analysis of QS showed that 91.7% of A. baumannii harbored AHL gene encoding N-acyl homoserine lactone hydrolase at amplified size 498bp, while 83.3% of A. baumannii isolates could harbor abaI gene encoding N-acyl homoserine lactone synthase at amplified size 382bp. Conclusion: it could be concluded that A. baumannii isolates were capable to produce biofilm and control it using quorum sensing genes expression. This would raise the concern of dissemination of chronic infections among hospitalized patients in two cities: Baghdad and Al-Najaf.

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References

- Lee H. W., Koh Y. M., Kim J., Lee J.C., Lee Y.C., Seol S.Y., et al. Capacity of multidrug-resistant clinical isolates of Acinetobacter baumannii to form biofilm and adhere to epithelial cell surfaces. Clinical Microbiology and Infection 2008; 14 (1): 49-54.

- Araujo C. D., Balestrino D., Roth L., Charbonnel N., Forestier C. Quorum sensing affects biofilm formation through lipopolysaccharide synthesis in Klebsiella pneumonia. Research in Microbiology 2010; 1(10):1-10.

- Hassan A.,Usman J., Kaleem F., Omair M., Khalid A., and Iqbal M. Evaluation of different detection methods of biofilm formation in the clinical isolates. Braz J Infect Dis 2011; 15(4): 305-311.

- Prashanth K., Vasanth T., Saranathan R., Makki A. R., and Pagal S. Antibiotic Resistance, Biofilms and Quorum Sensing in Acinetobacter Species, Antibiotic Resistant Bacteria - A Continuous Challenge in the New Millennium, Dr. Marina Pana (Ed.) 2012, ISBN: 978-953-51 0472-8.

- Chan K., Cheng H. J., Chen J. W., Yin W. F., and Ngeow Y. F. Tandem Mass Spectrometry Detection of Quorum Sensing Activity in Multidrug Resistant Clinical Isolate Acinetobacter baumannii. Scientific World Journal 2010:1-6

- Howard A., O’Donoghue M., and Sleator R. D. Acinetobacter baumannii. Virulence 2012;3(3):243-250.

- Clemmer K. M., Bonomo R. A., and Rather P. N. Genetic analysis of surface motility in Acinetobacter Baumannii. Microbiology 2011; 157:2534–2544.

- M’hamedi I, Hassaine H, Bellifa S., Lachachi M, Terki I.,K., and Djeribi R. Biofilm formation by Acinetobacter baumannii isolated from medical devices at the intensive care unit of the University Hospital of Tlemcen (Algeria).African Journal of Microbiology Research; 8 (3) 2014: 1-14.

- Aziz R.A.R., Al-Jubori S.S. Molecular Analysis of Genetic Elements Responsible for XDR in Highly Successful Pathogen Acinetobacter Baumannii Isolated from Clinical Samples of Iraqi Patients. Journal of Global Pharma Technology 2017; 04(9):26-39.

- O’Toole G. A. Microtiter dish biofilm formation assay. JoVE. 2011; 47:1-2.

- O’Toole, G. A., and Kolter, R. Flagellar and twitching motility are necessary for Pseudomonas aeruginosa biofilm development. Mol. Microbiol 1998; 30:295–304.

- Fischer E. R., Hansen B. T., Nair V., Hoyt F. H., and Dorward D. W. Scanning Electron Microscopy. Curr Protoc Microbiol 2012:1-76.

- Ajao A. O, Robinson G., Lee M. S., and Ranke T. D. Comparison of culture media for detection of Acinetobacter baumannii in surveillance cultures of critically-ill patients. Eur J Clin Microbiol Infect Dis.2011; 30(11): 1425–1430.

- Manikam K., Karlowsky J. A., Adam H., and Pang. CHROMagar Orientation Medium Reduces Urine Culture Workload. Journal of Clinical Microbiology2013; 51(2):1179–1183.

- Sanchez C. J., Mende K., Beckius M. L., Akers K. S. , Romano D. R., Wenke J. C., et al. Biofilm formation by clinical isolates and the implications in chronic infections. BMC Infectious Diseases 2013; 13 (47): 1-12.

- Farahani A., Mohajeri P. and Abbasi H. Comparison of Different Phenotypic and Genotypic Methods for the Detection of Methicillin-Resistant Staphylococcus Aureus. N Am J Med Sci. 2013; 5(11): 637–640.

- Ali R., Al-Achkar K., and Al-Mariri A. Role of Polymerase Chain Reaction (PCR) in the detection of antibiotic-resistant Staphylococcus aureus. The Egyptian Journal of Medical Human Genetics 2010;15:293–298.

- Khosravi A., Sadeghi P., and Sheikhi. Molecular Methods for Identification of Acinetobacter Species by Partial Sequencing of the rpoB and 16S rRNA Genes. Journal of Clinical and Diagnostic Research 2015; 9(7): DC09-DC13.

- Rodr´guez-Ban˜o J., Martı´S., Soto S., Ferna´ndez-Cuenca F., Cisneros J. M., Pacho´n J., et al. Biofilm formation in Acinetobacter baumannii: associated features and clinical implications. Clinical Microbiology and Infection 2008, 14 (3):1-13.

- Anbazhagan D, Mansor M, Yan GOS, Yusof MY, Hassan H, et al. Detection of Quorum Sensing Signal Molecules and Identification of an Autoinducer Synthase Gene among Biofilm Forming Clinical Isolates of Acinetobacter spp. PLoS ONE 7(7) 2012:1-12.

- Gentile V., Frangipani E., Bonchi C., Minandri F., Runci F. et al. Iron and Acinetobacter baumannii Biofilm Formation. Pathogens 2014; 3:704-719.

- Abdi-Ali A., Hendiani S., Mohammadi P., Gharavi S. Assessment of Biofilm Formation and Resistance to Imipenem and Ciprofloxacin among Clinical Isolates of Acinetobacter baumannii in Tehran. Jundishapur J Microbiol 2014; 7(1): 1-5.

- Qi H. L., Zhang C., and Song H. Relationship between Antibiotic Resistance, Biofilm Formation, and Biofilm-Specific Resistance in Acinetobacter baumannii. Frontiers in Microbiology 2016; 7:1-10.

- Babapour E., Haddadi A., and Mirnejad R. Biofilm formation in clinical isolates of nosocomial Acinetobacter baumannii and its relationship with multidrug resistance. Asian Pac J Trop Biomed 2016; 6(6): 528–533.

- Mussi M. Gaddy J. A., Arivett B., and Actis L. A. The Opportunistic Human Pathogen Acinetobacter baumannii Senses and Responds to Light. Journal of Bacteriology 2010; 192(24): 6336–6345.

- Eijkeikamp B, Stroeher U. H., Hassan K. A., and Paulsen I. T. Adherence and motility characteristics of clinical Acinetobacter baumannii isolates. FEMS Microbiol Lett 2011; 323: 44–51.

- Stanton A. R. Assessment of motility and hemolytic activity in clinical isolates of Acinetobacter baumannii from University of Kentucky hospital, Lexington, KY. Online Theses and Dissertations 2012:1-134.

- McQueary C. M., Kirkup B. C., and Zurawski D. V.Extracellular stress and lipopolysaccharide modulate Acinetobacter baumannii surface-associated motility. Journal of Microbiology 2012; 50(3): 434–443.

- Harding C., Tracy E., Carruthers M., Munson R. Acinetobacter baumannii Strain M2 Produces Type IV Pili Which Play a Role in Natural Transformation and Twitching Motility but Not Surface-Associated Motility. mBio 2013; 4(4):1-13.

- Kostoulias X., Murray G., and Peleg A. Impact of a Cross-Kingdom Signaling Molecule of Candida albicans on Acinetobacter baumannii Physiology. Antimicrob Agents Chemother 2016; 60:161–167.

- Vijayakumar S., Rajenderan S., and Biswas I. Biofilm Formation and Motility Depend on the Nature of the Acinetobacter baumannii Clinical Isolates. Front. Public Health 2016; 4(105):1-9.

- Brossard K.A., and Campagnari A., A. The Acinetobacter baumannii Biofilm Associated Protein Plays a Role in Adherence to Human Epithelial Cells. Infection and Immunity 2011; 80(1): 228–233.

- Sahu P. K., Pavithra S., Amrita M., and Chopade. Characterization of eDNA from the Clinical Strain Acinetobacter baumannii AIIMS 7 and Its Role in Biofilm Formation. The ScientificWorld Journal 2013:1-11.

- Basak S., Rajurkar M., and Mallick S. K. (2013). Biofilms: A Challenge to Medical Fraternity in Infection Control. Infection Control:57-74.

- Bandeira M. , Carvalho P.A., Duarte A., and Jordao L. Exploring Dangerous Connections between Klebsiella pneumoniae Biofilms and Healthcare-Associated Infections. Pathogens 2014; 3:720-731.

- Haley K., Blanz E., Kirk L., Gaddy J. Bacterial pathogenesis: Using high-resolution microscopy techniques to study host-pathogen interactions. Microscopy: advances in scientific research and education (A. Méndez-Vilas, Ed.) 2014.

- Narayanan A., Nair M. S., Venkitanarayanan K., and Amalaradjou M. A. R. Inactivation of Acinetobacter baumannii Biofilms on Polystyrene, Stainless Steel, and Urinary Catheters by Octenidine Dihydrochloride. Frontiers in Microbiology 2016;(7):1-9.

- Niu C., Clemmer K. M., and Rather P. Isolation and Characterization of an Autoinducer Synthase from Acinetobacter baumannii. Journal of Bacteriology 2008; 190(9):3386–3392.

- Gonzalez U., Pharm D., and Spencer J. Aminoglycosides: A Practical Review. Am Fam Physician 1998;58(8):1811-1820.

- He X., Lu F., Yuan F., Jiang D., Zho P. et al. Biofilm Formation Caused by Clinical Acinetobacter baumannii Isolates Is Associated with Overexpression of the AdeFGH Efflux Pump. Antimicrobial Agents and Chemotherapy 2015; 59(8): 4817-4825.

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