Effect of Culture Media and Incubation Temperature on Growth of Selected Strains of Francisella Tularensis

Abstract

The rate and amount of growth of 4 field isolates and reference strain ATCC 6223 of Francisella tularensis were evaluated on isolation media with 2 different agar bases and with different supplements and incubated at 25 C, 35 C, and 42 C. Biochemical reactions on conventional differential media with and without cysteine were evaluated. Two of the field isolates and the reference strain were F. tularensis subspecies tularensis (formerly biovar tularensis or Type A), and 2 isolates were subspecies holarctica (formerly subspecies palaearctica or Type B). Bacto cystine heart blood agar supplemented with 1% hemoglobin, glucose cystine heart blood agar, and brain-heart infusion blood agar supported good growth of all 4 field strains, with the most luxuriant growth occurring on Bacto cystine heart blood agar with hemoglobin. Heart infusion blood agar and trypticase soy blood agar supported growth of the field isolates, although growth was diminished and delayed. Strain 6223 was distinctly fastidious and failed to grow on heart infusion or trypticase soy blood agars. Growth of strain 6223 was best on Bacto cystine heart blood agar with hemoglobin. The agar base did not affect growth unless the supplements became limiting, in which case Bacto agar base generally supported growth better than BiTek agar base. Incubation at 35 C was optimum for all 5 strains. Growth at 42 C was slow, with the greatest decrease in the rate and amount of growth occurring with field isolates of F. tularensis subspecies tularensis. Strain 6223 did not grow at 25 C, and the 4 field isolates grew slowly at the lower temperature. The addition of cysteine did not alter any biochemical reactions with the exception of decreasing the detection time of H2S by subpsecies holarctica.