Detection of PgaABCD Proteins in Biofilm Producing Acinetobacter Baumannii Isolates

Authors

  • Esraa Eissa Rheama Biology Department, College of Science, Mustansiriyah University, 10052 Baghdad, IRAQ.
  • Susan Abdulrahman Ibrahim Biology Department, College of Science, Mustansiriyah University, 10052 Baghdad, IRAQ. https://orcid.org/0000-0002-5250-0512

DOI:

https://doi.org/10.23851/mjs.v33i5.1320

Keywords:

pgaABCD locus, PNAG, Acinetobacter baumannii, Biofilm, poly-1,6-N-acetylglucosamine

Abstract

Acinetobacter baumannii is a common cause of nosocomial infections. This bacterium is able to survive in hostile environments (desiccation, antimicrobial therapies, nutrient unavailability) beside colonization biotic and abiotic surfaces and form biofilm in hospitals and long-term care institutions. Biofilm is a three-dimensional structure of a multicellular complex in which the cells are embedded in an Extracellular Polymeric Substance (EPS) that was produced by the organism itself. Biofilm matrix contains proteins, ions, nucleic acids, and polysaccharide polymers. The main factor that leads to biofilm formation in A. baumannii is the creation of the exopolysaccharide poly-1,6-N-acetylglucosamine (PNAG), which is a virulence factor required for adhesion and aggregation by many Gram-negative bacteria. Many studies indicated that PNAG is an important factor to keep the integrity of A. baumannii biofilms in a more dynamic and stressful environment. A cluster of four genes (pgaA, pgaB, pgaC, and pgaD) are responsible for PNAG, which is encoded by pgaABCD locus. Therefore, the aim of this work was to detect proteins encoded by this locus using SDS-PAGE gel in A. baumannii isolates. Using PCR analysis to detect 16S rRNA concluded that all 24 isolates showed a positive amplification with 242bp. Acinetobacter baumannii clinical isolates showed high resistance percentage to ampicillin-sulbactam (AMS) and ceftazidime (CAZ) with 100% and 91% respectively. Furthermore, the isolates showed 83.3% for cefepime (FEP) and 70.8% for amikacin (AK), while the isolates showed a variable resistance percentage toward other antibiotics. By using Congo red method indicated that 66.7% were positive to produce biofilm and 33.3 were non-forming biofilm. Protein sequences alignment showed 99%, 99%, 100% and 99% identity for PgaA, PgaB, PgaC and PgaD respectively with protein GenBank database.

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References

L. V. Bentancor, J. M. O'Malley, C.Bozkurt-Guzel, G. B. Pier, and T. Maira-Litrán, "Poly- n -acetyl-β-(1-6)-glucosamine is a target for protective immunity against Acinetobacter baumannii infections," Infection and Immunity, vol. 80, no. 2, pp. 651-656, 2012.

CrossRef | PubMed

G. Eason, B. Noble, and I. N. Sneddon, On certain integrals of Lipschitz-Hankel type involving products of Bessel functions, vol. A247, London: Phil. Trans. Roy. Soc, 1955, pp. 529-551. M.-P. S. L and W. R. A, "Acinetobacter infection," New England Journal of Medicine, vol. 358, pp. 1271-1281, Mar. 2008.

CrossRef

B. Aarabi, "Comparative study of bacteriological contamination between primary and secondary exploration of missile head wounds," Neurosurgery, vol. 20, no. 4, pp. 610-616, 1987.

CrossRef | PubMed

O. Oncul, F. Yüksel, H. Altunay, C. Açikel, B. Çeliköz, and Ş. Çavuşlu, "The evaluation of nosocomial infection during 1-year-period in the Burn Unit of a training hospital in Istanbul, Turkey," Burns, vol. 28, no. 8, pp. 738-744, 2002.

CrossRef | PubMed

M. Maegele, S. Gregor, E. Steinhausen, B. Bouillon, M. M. Heiss, W. Perbix, F. Wappler, D. Rixen, J. Geisen, B. Berger-Schreck, and R. Schwarz, "The long-distance tertiary air transfer and care of tsunami victims: Injury pattern and microbiological and psychological aspects*," Critical Care Medicine, vol. 33, no. 5, pp. 1136-1140, 2005.

CrossRef | PubMed

N. E. Aronson, J. W. Sanders, and K. A. Moran, "Emerging infections: In harm's way: Infections in deployed American military forces," Clinical Infectious Diseases, vol. 43, no. 8, pp. 1045-1051, 2006.

CrossRef | PubMed

J. H. Calhoun, C. K. Murray, and M. M. Manring, "Multidrug-resistant organisms in military wounds from Iraq and Afghanistan," Clinical Orthopaedics & Related Research, vol. 466, no. 6, pp. 1356-1362, 2008.

CrossRef | PubMed

J. H. Calhoun, C. K. Murray, and M. M. Manring, "Multidrug-resistant organisms in military wounds from Iraq and Afghanistan," Clinical Orthopaedics & Related Research, vol. 466, no. 6, pp. 1356-1362, 2008.

CrossRef | PubMed

S. R. Lockhart, M. A. Abramson, S. E. Beekmann, G. Gallagher, S. Riedel, D. J. Diekema, J. P. Quinn, and G. V. Doern, "Antimicrobial resistance among gram-negative bacilli causing infections in intensive care unit patients in the United States between 1993 and 2004," Journal of Clinical Microbiology, vol. 45, no. 10, pp. 3352-3359, 2007.

CrossRef | PubMed

N. A. Al-Sweih, M. A. Al-Hubail, and V. O. Rotimi, "Emergence of tigecycline and colistin resistance inacinetobacterspecies isolated from patients in Kuwait hospitals," Journal of Chemotherapy, vol. 23, no. 1, pp. 13-16, 2011.

CrossRef | PubMed

R. A. Bonomo and D. Szabo, "Mechanisms of multidrug resistance in Acinetobacter species and pseudomonas aeruginosa," Clinical Infectious Diseases, vol. 43, no. Supplement_2, 2006.

CrossRef | PubMed

D. Mack, W. Fischer, A. Krokotsch, K. Leopold, R. Hartmann, H. Egge, and R. Laufs, "The intercellular adhesin involved in biofilm accumulation of Staphylococcus epidermidis is a linear beta-1,6-linked glucosaminoglycan: Purification and structural analysis," Journal of Bacteriology, vol. 178, no. 1, pp. 175-183, 1996.

CrossRef | PubMed

Maira-Litrán Tomás, A. Kropec, C. Abeygunawardana, J. Joyce, G. Mark, D. A. Goldmann, and G. B. Pier, "Immunochemical properties of the staphylococcal poly- n -acetylglucosamine surface polysaccharide," Infection and Immunity, vol. 70, no. 8, pp. 4433-4440, 2002.

CrossRef | PubMed

X. Wang, J. F. Preston, and T. Romeo, "The pgaABCD locus of escherichia coli promotes the synthesis of a polysaccharide adhesin required for biofilm formation," Journal of Bacteriology, vol. 186, no. 9, pp. 2724-2734, 2004.

CrossRef | PubMed

C. Darby, J. W. Hsu, N. Ghori, and S. Falkow, "Plague bacteria biofilm blocks food intake," Nature, vol. 417, no. 6886, pp. 243-244, 2002.

CrossRef | PubMed

A. H. Choi, L. Slamti, F. Y. Avci, G. B. Pier, and Maira-Litrán Tomás, "The pgaABCD locus of acinetobacter baumannii encodes the production of poly-β-1-6- n -acetylglucosamine, which is critical for biofilm formation," Journal of Bacteriology, vol. 191, no. 19, pp. 5953-5963, 2009.

CrossRef | PubMed

Y. Itoh, J. D. Rice, C. Goller, A. Pannuri, J. Taylor, J. Meisner, T. J. Beveridge, J. F. Preston, and T. Romeo, "Roles of pgaabcd genes in synthesis, modification, and export of the escherichia coli biofilm adhesin poly-β-1,6- n -acetyl- d -glucosamine," Journal of Bacteriology, vol. 190, no. 10, pp. 3670-3680, 2008.

CrossRef | PubMed

K. Vanbroekhoven, A. Ryngaert, P. Wattiau, R. Ã. Mot, and D. Springael, "Acinetobacter diversity in environmental samples assessed by 16S rrna gene pcrâ€'dgge fingerprinting," FEMS Microbiology Ecology, vol. 50, no. 1, pp. 37-50, 2004

CrossRef | PubMed

L. Chen, H. Li, H. Wen,B. Zhao, Y. Niu, Q. Mo and Wu, Y. "Biofilm formation in Acinetobacter baumannii was inhibited by PAβN while it had no association with antibiotic resistance". Microbiology Open, 9(9), e1063, 2020.

CrossRef

A.P. Magiorakos, A. Srinivasan, R. B. Carey, Y. Carmeli, M. E. Falagas, C. G. Giske, S. Harbarth, J. F. Hindler, G. Kahlmeter, B. Olsson-Liljequist, D. L. Paterson, L. B. Rice, J. Stelling, M. J. Struelens, A. Vatopoulos, J. T. Weber, and D. L. Monnet, "Multidrug-resistant, extensively drug-resistant and Pandrug-resistant bacteria: An international expert proposal for interim standard definitions for acquired resistance," Clinical Microbiology and Infection, vol. 18, no. 3, pp. 268-281, 2012.

CrossRef | PubMed

"M100ED32: Performance standards for antimicrobial susceptibility testing, 32nd edition," Clinical & Laboratory Standards Institute. [Online]. Available: https://www.clsi.org/standards/products/microbiology/documents/m100/. [Accessed: 22-Aug-2022].

A. Hassan, J. Usman, F. Kaleem, M. Omair, A. Khalid, and M. Iqbal, "Evaluation of different detection methods of biofilm formation in the clinical isolates," Brazilian Journal of Infectious Diseases, vol. 15, no. 4, pp. 305-311, 2011.

CrossRef | PubMed

U. K. LAEMMLI, "Cleavage of structural proteins during the Assembly of the head of bacteriophage T4," Nature, vol. 227, no. 5259, pp. 680-685, 1970.

CrossRef | PubMed

L. B. Rice, "Federal funding for the study of antimicrobial resistance in nosocomial pathogens: No eskape," The Journal of Infectious Diseases, vol. 197, no. 8, pp. 1079-1081, 2008.

CrossRef | PubMed

A. Y. Peleg, H. Seifert, and D. L. Paterson, "acinetobacter baumannii : Emergence of a successful pathogen," Clinical Microbiology Reviews, vol. 21, no. 3, pp. 538-582, 2008.

CrossRef | PubMed

M. A., Rizk & M. El-Khier, "Aminoglycoside resistance genes in Acinetobacter baumannii clinical isolates," Clinical Laboratory, vol. 65, no. 07/2019, 2019.

CrossRef | PubMed

Z. Li, Z. Ding, Y. Liu, X. Jin, J. Xie, T. Li, Z. Zeng, Z. Wang, and J. Liu, "Phenotypic and genotypic characteristics of biofilm formation in clinical isolates of Acinetobacter baumannii," Infection and Drug Resistance, vol. Volume 14, pp. 2613-2624, 2021.

CrossRef | PubMed

S. Garneau-Tsodikova and K. J. Labby, "Mechanisms of resistance to aminoglycoside antibiotics: Overview and perspectives," MedChemComm, vol. 7, no. 1, pp. 11-27, 2016.

CrossRef | PubMed

E. Babapour, A. Haddadi, R. Mirnejad, S. A. Angaji, and N. Amirmozafari, "Biofilm Formation in clinical isolates of nosocomial Acinetobacter baumannii and its relationship with multidrug resistance," Asian Pacific Journal of Tropical Biomedicine, vol. 6, no. 6, pp. 528-533, 2016.

CrossRef

F. Runci, C. Bonchi, E. Frangipani, D. Visaggio, and P. Visca, "Acinetobacter baumannii biofilm formation in human serum and disruption by Gallium," Antimicrobial Agents and Chemotherapy, vol. 61, no. 1, 2017.

CrossRef | PubMed

I. A. Raheem, S. S. Hussain, R. H. Essa, A.-J. K. Atia, and F. R. Abdul, "Biological activity of new hydantoin derivatives on Acinetobacter baumannii biofilm formation isolated from clinical sources," Journal of University of Babylon for Pure and Applied Sciences, vol. 26, no. 10, p. 71-79, 2018.

L. G. H. Fajir, A. E. Al-Niaame, and N. N. Hussein, "Detection of biofilm formation and its related with aminoglycoside resistance in Acinetobacter baumannii isolates, isolated from some Baghdad City Hospitals," Eur J Biomed Pharm Sci, vol. 4, no. 1, pp. 84-91, 2017.

D. J. Freeman, F. R. Falkiner, and C. T. Keane, "New method for detecting slime production by coagulase negative staphylococci" Journal of Clinical Pathology, vol. 42, no. 8, pp. 872-874, 1989.

CrossRef | PubMed

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Key Dates

Published

25-02-2023

How to Cite

[1]
E. E. . Rheama and S. A. . Ibrahim, “Detection of PgaABCD Proteins in Biofilm Producing Acinetobacter Baumannii Isolates”, Al-Mustansiriyah J. Sci., vol. 33, no. 5, pp. 78–87, Feb. 2023, doi: 10.23851/mjs.v33i5.1320.

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