Effect of culture medium on morphology and virulence of Legionella pneumophila serogroup 1.

In a preliminary study, the preparation of a modified charcoal yeast extract by predialysis of yeast extract (CDYE) allowed us to obtain short non filamentous forms of Legionella pneumophila ser 1 (Philadelphia) found to be more virulent in the chick embryo than the long forms grown on conventional media. We confirmed these findings in guinea pigs inoculated by either intraperitoneal injection or aerosol inhalation. LD50s were calculated using the method of Reed and Muench. Survival curves were established using Liddell's method. If for chick embryo the most virulent organisms were those derived from yolk sac culture, organisms grown on CDYE agar were more virulent than those grown on the other media. There was a significant positive correlation between the mean length of the bacilli and the log 10 of the LD50 (r = 0.96; 0.02 less than p less than 0.05). For guinea pigs by either intraperitoneal injection or inhalation we confirmed that the bacteria cultured on CDYE were more virulent than those grown on other solid media. Thus for the guinea pig inoculated intraperitoneally, the LD50s of the CDYE and BCYE cultures were 1.4 X 10(7) and greater than 3 X 10(9) CFU, respectively. The mortality of guinea pigs inoculated by aerosol with CDYE cultures was significantly higher than that of guinea pigs infected with BCYE cultures using suspensions of 10(8) and 10(9) CFU/ml (p less than 0.01) and 10(10) CFU/ml (p less than 0.05).