Toxic activity of Bacillus sphaericus SSII-1 for mosquito larvae

Using larvae of the mosquito Culex pipiens var. quinquefasciatus as a bioassay system, we have verified an earlier proposal that pathogenicity of Bacillus sphaericus SSII-1 is a toxin-mediated rather than an infectious process. Chloroform or ultraviolet-light treatments that decreased the viable count of SSII-1 cells by 4 or 5 logs did not significantly alter the ability of the bacterial cells to kill larvae. Three lines of evidence indicated that toxic activity was not related to sporulation: (i) cells grown in either a complex or a defined medium were toxic at all ages; (ii) when supplemental Mn2+ was excluded from a complex medium, the culture yielded few spores but was of equal toxicity to a culture containing many spores; and (iii) several early blocked oligosporogenous mutants were isolated that had toxic activities comparable to that of the parent. The toxin was shown to be relatively unstable because activity was destroyed by heat and decreased by refrigeration, a freeze-thaw cycle, or two methods of cell breakage. Thin sections of SSII-1 cells did not reveal the presence of any inclusion body that might be related to toxicity.