Optimization of some environmental and nutritional conditions using microtiter plate for Pseudomonas aeruginosa biofilm formation

Abstract

One of the most important virulence factors in Pseudomonas aeruginosa is biofilm formation, as it works as a barrier for entering antibiotics into the bacterial cell. Different environmental and nutritional conditions were used to optimize biofilm formation using microtitre plate assay by P. aeruginosa. The low nutrient level of the medium represented by tryptic soy broth (TSB) was better in biofilm formation than the high nutrient level of the medium with Luria Broth (LB). The optimized condition for biofilm production at room temperature (25 °C) is better than at host temperature (37 °C). Moreover, the staining with 0.1% crystal violet and reading the biofilm with wavelength 360 are considered essential factors in increasing the productivity of biofilm formation in P. aeruginosa. Finally, we highly recommended using these optimized microtitre plate assays to assess biofilm formation in P. aeruginosa.

Study on tssC1 Gene Mediating Biofilm Antibiotics Resistance of Pseudomonas aeruginosa

“…Bacterial suspension was prepared by mixing 20 μl (Bacterial culture was adjusted to the turbidity of a 1 McFarland standard) of each 20 P. aeruginosa isolates with 180 μl tryptic soy broth. Later 0.25% glucose was added to the suspension and was then kept rest for 24 h at 37 0 C [17]. The suspension was then loaded into 96 wells of microtitre plate and incubated at 37 0 C for 48 h. Control was also loaded in the wells which contained broth without inoculation.…”