QUANTITATIVE AND QUALITATIVE ASSAYS OF BACTERIAL BIOFILM PRODUCED BY Pseudomonas aeruginosa AND Klebsiella spp.
|Journal of University of Anbar for Pure Science|
|Article 3, Volume 2, Issue 3, December 2008, Pages 6-13 PDF (295.14 K)|
|Document Type: Research Paper|
|Shaymaa H. Moteeb*|
|College of Dentistry, University of Anbar|
|The aim of this study was to detect biofilm formation by study isolates of Pseudomonas aeruginosa and Klebsiella spp. qualitatively and quantitatively. Twenty-five isolates were taken from patients admitted to Ramadi General Hospital were included in this study. Qualitative biofilm formation assays (tube method and Foley-catheter assay) and quantitative assay by spectrophotometric method with ELISA reader were achieved under two experimental conditions. In tube method, the result showed that out of 8 (32%) isolates of Pseudomonas aeruginosa and 17 (68%) isolates of Klebsiella species, the biofilm were produced in 7 (87.5%), and 14(82.35%) respectively. while, in foley catheter method, the biofilm were produced on the surfaces of the catheters in all pseudomonal isolates (100%), and 15(88.23%)of klebsiella isolates respectively. In the spectrophotometric method, the results showed that out of 17 isolates of Klebsiella spp., all isolates were produced biofilm strongly in the glucose supplemented media while 15 (88.23%) of them were produced biofilm strongly in the glucose non-supplemented media and 2 (11.76%) isolates were weak biofilm producers in the glucose non-supplemented media. With regard to Pseudomonas aeruginosa, the results showed that all pseudomonal isolates, which submitted to this study, were produced biofilm strongly in the glucose supplemented and non-supplemented brain heart infusion broth. The study concluded that all isolates of Pseudomonas aeruginosa and Klebsiella spp. were produced biofilm qualitatively by both techniques. Further, the spectrophotometric method was an accurate method for detection the bacterial adherence to the surface of Microtiter plates. Further more, biofilm production was not affected by this factor in both of Pseudomonas aeruginosa and Klebsiella spp.|
|Bacterial biofilm; Pseudomonas aeruginosa; Klebsiella spp|
1-Costerton, J.W., Cheng, K.J., Geesey, G.G., Ladd, T.I., Nickel, J.C., Dasgupta, M. and Marrie, T.J. (1987). Bacterial biofilms in nature and disease. Ann. Rev. Microbiol., 41: 435-464.
2- Davey, M.E., and O’Toole, G.A., (2000). Microbial biofilms: from ecology to molecular genetics. Microbiol. Mol. Biol. Rev. 64, 847–867.
3-Costerton, J.W., Stewart, P.S., and Greenberg, E.P., (1999).Bacterial biofilms: a common cause of persistent infections. Science 284, 1318–1322.
4-Donlan, R.M., and Costerton, J.W., (2002). Biofilms: survival mechanisms of clinically relevant microorganisms. Clin. Microbiol. Rev. 15, 167–193.
5-Dunne, W.M., (2002).Bacterial adhesion: seen any good biofilm lately? Clin. Microbiol. Rev15, 155–166.
6-Mikkelsen, H., Duck, Z., K. S. Lilley, K. S, and M. Welch.(2006). Interrelationships between Colonies, Biofilms, and Planktonic Cells of Pseudomonas aeruginosa. agenetics. Microbiol. Mol. Biol. Rev. 64, 847–867.
7-Baron, E. J., Peterson, L.R., Finegold, S.M. (1994). Method for testing antimicrobial effectiveness. In: Diagnostic Microbiology, Bail and Scott's. Toronto: C.V. Mobsy company., 9th ed: 168-193.
8-Yassien, M., and khardori, N.(2001).Interaction between biofilm formed by staphylococcus epidermidis &quinolones. Diagnostic microbiology and infectious disease.40 (volume):79-89.
9-Mireles, J.R., Toguchi, A and Harshy, R.M. (2001). Salmonella enterica serovar typhimurium swarming mutant with altered biofilm forming abilities: Surfactin inhibits biofilm formation. J. Bacteriol., 183: 5848-5854.
10-Arif, S.K. (2006). Catheter associated bacteriuria and biofilm study. Ph.D thesis. College of Medicine, University of Al-Mustansiryia.
11- Stepanovic, S., Djukic, V., Djordjevic, V. and Djukic, S. (2003). Influence of the incubation atmosphere on the production of biofilm by staphylococci. Clin Microbiol. Infect, 9: 955-959.
12-Christensen, G.D., Simpson, W.A., Younger, J.J., Baddour, L.M., Barrett, F.F., Melton, D.M., and Beachey, E.H. (1985). Adherence of Coagulase negative staphylococci to plastic tissue culture plates: A quantitative model for the adherence of staphylococci to medical devices. J. Clin. Microbiol., 22: 996-1006.
13- Katsikogianni, M. and Missirlis, Y.F. (2004). Concise review of mechanisms of bacterial adhesion to biomaterials and of techniques used in estimating bacteria-material interactions. Europ. Cell Mat., 8: 37-57.
14- Donlan, R.M. (2002). Biofilms: Microbial Life on Surfaces. Emerg. Infect. Dis.8: 881-890.
15-Jones, B.V., Mahenthiralingam, E., Sabbuba, N.A., Stickler, D.J. (2005). Role of swarming in the formation of crystalline Proteus mirabilis biofilms on urinary catheters. J. Med. Microbiol., 54: 807-813.
16-Mathur, T., Singhal, S., Khan, S., Upadhyay, D.J., Fatma, T. and Rattan, A. (2006). Detection of biofilm formation among the clinical isolates of Staphylococci: An evaluation of three different screening methods. Ind. J. Med. Microbiol., 24: 25-29.
17-Stepanovic, S., Vukovic, D., Dakic, I., Savic, B., Svabic-Vlahovic, M. (2000). A modified microtiter plate test for quantification of staphylococcal biofilm formation. J. Microbiol. Meth., 40: 175-179.
Article View: 81
PDF Download: 53