Double-J (DJ) Stenting Lab Report

Double-J (DJ) stenting is the commonest procedure performed in urology. Patients with stent, develop stent infections and stent-related symptoms. Use of antibiotics in patients with DJ stent is rampant. No uniform data is there to prove advantage of antibiotic prophylaxis. Colonial way of life of microorganisms, complex microbial assemblages are responsible for formation of biofilm. Both gram positive and gram negative bacteria produces biofilm. Biofilm is an important virulence factor and is main cause of many chronic infections and multidrug resistant strains resulting in treatment failure.
Microbially derived sessile communities containing cells that are attached to a substratum or to each other are called biofilms. They are embedded in
This is a quantitative test and is considered to be the gold-standard method for biofilm detection (13). Organisms isolated from fresh agar plates were inoculated in 10 mL of trypticase soy broth with 1% glucose. Broths were incubated at 370C for 24 hours. The cultures were then diluted 1:100 with fresh medium. Individual wells of sterile 96 well-flat bottom polystyrene tissue culture treated plates (Sigma-Aldrich, Costar, USA) were filled with 200 µL of the diluted cultures. The control organisms were also incubated, diluted and added to tissue culture plate. Negative control wells contained inoculated sterile broth. The plates were incubated at 370C for 24 hours. After incubation, gentle tapping of the wells were done to remove excess contents. The wells were washed four times with 0.2 mL of phosphate buffer saline (pH 7.2) to remove free floating bacteria. 2% sodium acetate was used for fixing the biofilm which was formed by bacteria adherent to the wells and is stained by crystal violet (0.1%). Excess stain was removed by using deionized water and plates were kept for
(8). This is a qualitative method for detection of biofilm. A loopful of test organisms was inoculated in 10 mL of trypticase soy broth with 1% glucose in test tubes. The tubes were incubated at 370C for 24 hours. Tubes were decanted and washed with phosphate buffer saline (pH 7.3) and incubated. Tubes were dried. Tubes were then stained with crystal violet (0.1%). Excess stain was washed with deionized water. Tubes were dried in inverted position. When a visible film lined the wall and the bottom of the tube, Biofilm formation was considered positive. Ring formation at the liquid interface is not indicative of biofilm