Pseudomonas Agar Medium For Detection of Fluorescein Medium 20 (In accordance with IP 2007)

Principle And Interpretation

Pseudomonas Agar (For Fluorescein) is based on the formula described by King et al (1) and modified as in the Indian Pharmacopoeia (2) for the detection of fluorescein production a water soluble, chloroform insoluble fluorescent pigment by Pseudomonas species and in microbial limit tests(3). Pseudomonas are ubiquitous in environment and is common causative agent of burn, skin and nosocomial infections. They are also common contaminant of pharmaceutical and cosmetics related preparations. Pseudomonas strains are reported to produce phenazine pigments like Pyocyanin- blue green redox-active secondary metabolite pigment, pyorubin-rust brown pigment, -oxyphenzine- a breakdown product of Pyocyanin, pyoverdin-a water soluble yellow green pigments also known as fluorescein.

This medium enhances the elaboration of fluorescein by Pseudomonas and inhibits the pyocyanin formation. The fluorescein pigment diffuses from the colonies of Pseudomonas into the agar and shows yellow fluorescent colouration. Some Pseudomonas strains produce small amounts of pyocyanin resulting in a yellow-green colouration. Peptic digest of animal tissue and pancreatic digest of casein provides the essential nitrogenous nutrients, carbon, sulfur and trace elements for the growth of Pseudomonas. These nutrients are also conducive to the production of fluroescein. Peptone and phosphorous in the medium enhances the production of pyoverdin/fluorescein pigment. Dipotassium phosphate buffers the medium while magnesium sulphate provides necessary cations for the activation of fluorescein production.

Salt concentration exceeding 2% affects pigment production. UV illumination may be bactericidal, so make sure that there is good growth before placing culture under UV light (3). The presence of Pseudomonas is confirmed by a positive oxidase test.