Studies on Efficient Methods for Detection of Legionella species from Environmental Water by use of its Selective Media

Abstract

To examine the optimal pH range for growth on media, growth of Legionella spp. on its selective media, BCYE alpha, WYO alpha and MWY agar media, in a pH range of 6.0-8.0 (at 0.5 intervals) was compared. The growth of two strains of L. pneumophila and one strain of L. micdadei on a WYO alpha agar supplemented with some selecting antimicrobial agents was markedly inhibited at all pH range except 6.0 and 7.0, suggesting a narrow optimal pH range for growth of these species compared to the BCYE alpha without selecting antimicrobioal agents. Vancomycin (VCM) added to the selective agar suppressed the growth of some Legionella spp. depending on the concentration. However, the extent of suppression was different among species and/or strains of Legionella spp. The selectivity for species other than Legionella spp. was also affected similarly by VCM concentration added to their media, suggesting that it is important to use proper amounts of the selecting antimicrobial agent depending on the species and/or strains of Legionella spp. or the other species in water samples. Amphothericin B (AMPH-B) added to a selective medium, MWY agar, in the concentration of 80 micrograms/ml hardly affected the growth of Legionella spp. examined, but effectively inhibited the growth of fungal strains identified as Aspergillus sp., Trichoderma sp., Scolecobasidium sp. and Mucor sp. which were isolated from cooling-tower water samples together with Legionella spp. Furthermore, the growth of a combination culture of one each of the 4 strains of isolated fungi and one each of the 3 strains of Legionella spp. was examined at various concentration of AMPH-B. Addition of AMPH-B to the selective medium at the concentration of 80 micrograms/ml suppressed the growth of spreading fungi, permitting the growth of Legionella spp. to allow efficient detection of the species.