Original Articles

Effect of Zinc Oxide Nanoparticle on the Expression of mrkA and fimA in Drug-Resistant Klebsiella pneumoniae

Abstract

Background:     Nanoparticles are a new generation of antimicrobials. Zinc oxide nanoparticles have attracted a great deal of interest in their medical applications. The aim of the present study was to investigate the effect of zinc nanoparticles on the expression of mrkA and fimA genes in drug-resistant K. pneumoniae. Methods:     A total of 30 clinical isolates of K. pneumoniae were collected from Sina hospital and all the isolates were identified by biochemical tests. Antimicrobial resistance pattern was determined by disk diffusion method. PCR method was used to investigate the presence of mrkA and fimA genes. Biofilm phenotypic test was performed and after conducting MIC test by micro dilution method, real-time PCR was used to study the effect of zinc oxide nanoparticle on the expression of fimA and mrkA genes. Results:     The highest resistance rate was against cefotaxime and ceftazidime antibiotics (67%). Twenty seven isolates harbored fimA gene while 24 isolates harbored mrkA gene. Five isolates were identified as strong biofilm producers. MIC values for zinc oxide was 2500 µg/ml in all five isolates. Results of real-time PCR showed that the expression levels of mrkA and fimA genes in isolates treated with zinc oxide decreased 8.5 and 9 fold, respectively, compared with the control. Conclusion:     This study suggests that zinc oxide can be a suitable candidate for the inhibition of the two studied virulence genes in K. pneumoniae.
1. DeLeo FR, Chen L, Porcella SF, Martens CA, Kobayashi SD, et al. Molecular dissection of the evolution of carbapenem-resistant multilocus sequence type 258 Klebsiella pneumoniaee. Proc Natl Acad Sci 2014; 111(13):4988–93.
2. Struve C, Bojer M, Krogfelt KA. Characterization of Klebsiella pneumoniaee type 1 fimbriae by detection of phase variation during colonization and infection and impact on virulence. Infect Immun 2008; 76(9):4055–65.
3. Podschun R, Ullmann U. Klebsiella spp. as nosocomial pathogens: epidemiology, taxonomy, typing methods, and pathogenicity factors. Clin microb rev 1998 Oct 1; 11(4):589-603.
4. Schurtz Sebghati TA, Korhonen TK, Hornick DB, et al. Characterization of the type 3 fimbrial adhesins of Klebsiella strains. Infect Immun 1998; 66(6):2887–94.
5. Ghasemian A, Mobarez AM, Peerayeh SN, et al. The association of surface adhesin genes and the biofilm formation among Klebsiella oxytoca clinical isolates. New Microbes New Infect 2019; 27:36–9.
6. Stoimenov PK, Klinger RL, Marchin GL, et al. Metal oxide nanoparticles as bactericidal agents. Langmuir 2002; 18(17):6679–86.
7. Leriche V, Chassaing D, Carpentier B. Behaviour of L. monocytogenes in an artificially made biofilm of a nisin-producing strain of Lactococcus lactis. Int J Food Microbiol 1999;51(2–3):169–82.
8. Madathil AN, Vanaja KA, Jayaraj MK. Synthesis of ZnO nanoparticles by hydrothermal method. In Nanophotonic materials IV 2007 Sep 17 (Vol. 6639, p. 66390J). SPIE.
9. Soumya KR, Snigdha S, Sugathan S, et al. Zinc oxide–curcumin nanocomposite loaded collagen membrane as an effective material against methicillin-resistant coagulase-negative Staphylococci. 3 Biotech 2017; 7(4):1-0.
10. Shakerimoghaddam A, Ghaemi EA, Jamalli A. Zinc oxide nanoparticle reduced biofilm formation and antigen 43 expressions in uropathogenic Escherichia coli. Iran J Basic Med Sci 2017; 20(4):451–6.
11. Bauer AW, Kirby WM, Sherris JC, et al. Antibiotic susceptibility testing by a standardized single disk method. Am J clin pathol 1966; 45:149-58..
12. O’Toole GA. Microtiter Dish Biofilm Formation Assay. J Vis Exp 2011; (47):1-0.
13. Xu L, Sun X, Ma X. Systematic review and meta-analysis of mortality of patients infected with carbapenem-resistant Klebsiella pneumoniaee. Ann Clin Microbiol Antimicrob 2017; 16(1):1-2.
14. Espitia PJP, Soares N de FF, Coimbra JS dos R, et al. Zinc oxide nanoparticles: synthesis, antimicrobial activity and food packaging applications. Food Bioproc Tech 2012:1447–64.
15. Vuotto C, Longo F, Balice M, et al. Antibiotic resistance related to biofilm formation in Klebsiella pneumoniaee. Pathogens 2014; 3(3):743–58.
16. Apondi OE, Oduor OC, Gye BK, et al. High prevalence of multi-drug resistant Klebsiella pneumoniaee in a tertiary teaching hospital in Western Kenya. African J Infect Dis 2016; 10(2):89–95.
17. Singla S, Harjai K, Chhibber S. Susceptibility of different phases of biofilm of Klebsiella pneumoniaee to three different antibiotics. J Antibiot (Tokyo) 2013; 66(2):61–6.
18. Samia B, Hafida H, Damien B, et al. Evaluation of biofilm formation of Klebsiella pneumoniaee isolated from medical devices at the University Hospital of Tlemcen, Algeria. African J Microbiol Res 2016; 7(49):5558–64.
19. Černohorská L, Votava M. Determination of minimal regrowth concentration (MRC) in clinical isolates of various biofilm-forming bacteria. Folia Microbiol (Praha) 2004; 49(1):75–8.
20. Niveditha S, Pramodhini S, Umadevi S, et al. The isolation and the biofilm formation of uropathogens in the patients with catheter associated urinary tract infections (UTIs). J Clin Diagnostic Res 2012; 6(9):1478–82.
21. Yang D, Zhang Z. Biofilm-forming Klebsiella pneumoniaee strains have greater likelihood of producing extended-spectrum β-lactamases. J Hosp Infect 2008; 68(4):369–71.
22. Cruz-Córdova A, Esteban-Kenel V, Espinosa-Mazariego K, et al. Pathogenic determinants of clinical Klebsiella pneumoniae strains associated with their persistence in the hospital environment. Bol Med Hosp Infant Mex 2014; 71(1):15-24
23. Alcántar-Curiel MD, Blackburn D, Saldaña Z, et al. Multi-functional analysis of Klebsiella pneumoniaee fimbrial types in adherence and biofilm formation. Virulence 2013; 4(2):129–38.
24. Lee JH, Kim YG, Cho MH, et al. ZnO nanoparticles inhibit Pseudomonas aeruginosa biofilm formation and virulence factor production. Microbiol Res 2014; 169(12):888–96.
25. Azam A, Ahmed AS, Oves M, et al. Antimicrobial activity of metal oxide nanoparticles against Gram-positive and Gram-negative bacteria: A comparative study. Int J Nanomedicine 2012; 7:6003–9.
Files
IssueVol 10 No 3-4 (2021) QRcode
SectionOriginal Articles
Keywords
Klebsiella pneumoniae Zinc Oxide Nanoparticle Real-Time PCR.

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
1.
Shivaee A, Kashani S, Mohammadzadeh R, Ebrahimi M. Effect of Zinc Oxide Nanoparticle on the Expression of mrkA and fimA in Drug-Resistant Klebsiella pneumoniae. J Med Bacteriol. 2021;10(3-4):1-10.