Influence of temperature on growth of Legionella pneumophila biofilm determined by precise temperature gradient incubator

Bacterial growth is influenced by several different culture conditions. Temperature is one of an essential component which regulates bacterial growth and their morphology. The influence of temperature on the length of bacteria was investigated in broth and on agar in a temperature range from 30.0°C to 47.0°C in 0.5°C steps using a newly developed temperature gradient incubator. The incubator is able to reach a set temperature within 2 h and maintain temperature as accurate as ±0.1°C of the set temperature. Three Legionella pneumophila serotype 1 strains were incubated for 48 h in BCYE-α agar at various temperatures ranging from 30.0°C to 48.0°C and length of bacteria grown at each temperature was microscopically measured. Ability of bacteria to multiply at a given temperature was also determined. L. pneumophila serotype 1 strains ATCC 33152, a clinical isolate Okinawa 02-001 were going to elongate to longer than 100 μm when cultured higher than at 39.5°C and at 41.5°C, respectively. Each strain was unable to multiply when cultured higher than at 44.2°C (ATCC 33152) or at 44.0°C (Okinawa 02-001). Those data would provide insights for establishing regulations in terms of maintaining hot water temperature in a facility where a circulating hot water supply-system is available and contamination with Legionella spp. is likely to happen.