Growth, respiration and survival of Legionella pneumophila at high temperatures.

The optimum temperature for multiplication of legionella strains in culture media is around 37 degrees C. The effect of high temperatures on the growth of strains isolated from various environments is poorly known. We studied the growth (cell multiplication, respiration) of clinical and environmental Legionella pneumophila strains in liquid media at intervals of 0.5 degrees C in the temperature range from 41.6 to 51.6 degrees C using a temperature gradient incubator. Cell multiplication and CO2 production decreased markedly with all the strains at temperatures above 44-45 degrees C. CO2 continued to be produced up to 51.6 degrees C even if cell multiplication generally stopped at around 48.4-50.0 degrees C. Thus, legionella retained its metabolic activity beyond the maximum temperature for cell multiplication. The CO2 production per bacterial cell (metabolic quotient, qCO2) increased with increasing temperature up to 45 degrees C, whereafter it decreased, the turning point being almost at the same at which the rate of cell multiplication decreased. The difference in qCO2 between the strains] may reflect their different physiological capacities for tolerating high temperatures.

Efficacy of three prevention strategies against legionella in cooling water systems.

The efficacy of three different prevention strategies on legionella in cooling systems was studied. The strategies were as follows: (1) water temperature was lowered; (2) water quality was improved; or (3) the system as disinfected with polyhexamethylene biguanidechloride (PHMB) biocide or with 2-bromo-2-nitropropane-1,3-diol (BNPD) biocide. Lowering of water temperature was the most effective method to reduce the concentration of legionella in cooling systems. Improving of water quality resulted in a transitory disinfection effect. The additions of PHMB or BNPD decreased the concentrations of both legionella and heterotrophic bacteria in cooling water. The effect of biocides, however, lasted at the most only a few months. If possible, lowering water temperature and improving the water quality should be the primary practices for controlling bacterial growth in cooling systems. Regular biocide treatments should be incorporated into the maintenance procedures if technical improvements cannot be done or if their efficiency is too low.

Isolation of Legionella from water samples using various culture methods.

The efficacy of a non-selective medium and two selective media were compared for the isolation of legionellas from water samples. The effect of acid wash treatment for decontamination of the water samples on the isolation frequency of legionellas was also studied. The 236 samples were taken from cooling, humidifying and drinking water systems; 21% were legionella-positive when inoculated directly on modified Wadowsky-Yee (MWY) medium and 26% were positive when concentrated (x 200) before cultivation on MWY or CCVC media. Inoculation on MWY medium after concentration followed by decontamination by the acid-wash technique gave the highest isolation frequency (31%). The lowest frequency (8%) was found with the non-selective BCYE alpha medium. An isolation frequency of 28% was achieved with the BCYE alpha medium after concentration and acid-wash treatment of the samples. Forty per cent of the samples were positive for legionellas when the results from all the culture methods were combined. Not all the legionella-positive samples were identified by a single culture method. Ninety-three of the 95 positive samples were detected with the two best combinations of three culture methods. The best culture method for detecting legionellas depended on the source of the water sample. Some water quality characteristics, like temperature and organic matter content, affected the isolation frequency of Legionella spp.

Growth of Legionella and other heterotrophic bacteria in a circulating cooling water system exposed to ultraviolet irradiation.

The effect of ultraviolet irradiation on the growth and occurrence of legionella and other heterotrophic bacteria in a circulating cooling water system was studied. Water of the reservoir was circulated once in 28 h through a side-stream open channel u.v. radiator consisting of two lamps. Viable counts of legionellas and heterotrophic bacteria in water immediately after the u.v. treatment were 0-12 and 0.7-1.2% of those in the reservoir, respectively. U.v. irradiation increased the concentration of easily assimilable organic carbon. In the u.v. irradiated water samples incubated in the laboratory the viable counts of heterotrophic bacteria reached the counts in reservoir water within 5 d. The increase in viable counts was mainly due to reactivation of bacterial cells damaged by u.v. light, not because of bacterial multiplication. Despite u.v. irradiation the bacterial numbers in the reservoir water, including legionellas, did not decrease during the experimental period of 33 d. The main growth of bacteria in the reservoir occurred in biofilm and sediment, which were never exposed to u.v. irradiation.