Development of a New Pseudomonas Agar Medium Containing Benzalkonium Chloride in Cetrimide Agar

Abstract

Members of the Pseudomonas family are commonly found in nature, some species are pathogenic for humans, as well as being resistant to multiple disinfectants. Various studies have revealed that benzalkonium chloride (BKC) has an inhibitory effect on many bacteria but it has no significant effect on Pseudomonas aeruginosa. Cetrimide agar medium is recommended for the isolation and enumeration of Ps. aeruginosa in food and environmental samples. However, there are claims that for some food factories and in particular the bottled water industry, the selectivity of this medium is not sufficient. The aim of the current research is the creation of a more selective medium for Ps. aeruginosa with BKC. A total of 28 isolates were isolated with Cetrimide agar from raw water samples and identified using biochemical tests and commercial identification kits. All the bacteria were inoculated in Cetrimide agar plates containing 0 - 625 μg/mL BKC. The Petri dishes were incubated at 37°C and 42°C for 24 h. The results showed that 375 μg/mL BKC was sufficient to suppress Burk. pseudomallei at both incubation temperatures. Ps. fluorescens-35 could not grow at 42°C at any concentration, including the control, and was suppressed at 500 μg/mL BKC. All the Ps. aeruginosa isolates and control strain were grown at both incubation temperatures at 375 μg/mL BKC concentration. In conclusion, the analysis of Ps. aeruginosa showed that the growth of accompanying flora may be suppressed by adding 375-μg/mL BKC into Cetrimide agar and incubating at an elevated temperature of 42°C.