Phenotypic and genetic diversity among Bacillus sphaericus strains isolated in Brazil, potentially useful as biological control agents against mosquito larvae.

Thirty mosquitocidal strains of Bacillus sphaericus isolated from different sources and localities in Brazil were characterized phenotypically and genetically to determine their relationship. Among the strains tested, 93.3% were shown to be resistant to lincomycin, 96.6% to novobiocin, 60% to chloramphenicol and all strains were resistant to streptomycin. Resistance to HgCl2, NiSO4.6H2O and CuSO4 was observed in 83.3, 86.6 and 100% of the strains, respectively. All strains were inhibited by the presence of CoSO4. Tolerance to ethanol and variable responses to different amounts of creolin, phenol and xylol was also observed. Amplification of DNA of each of 30 isolates using repetitive primers allowed the identification of 5 groups of similar strains in BOX-PCR and 8 groups in REP-PCR. Using cloned toxin genes from B. sphaericus as probes in hybridization studies, 83% of the strains studied hybridized to the bin probe and 90% to the mtx probe. A comparison of the 30 strains by similarity matrix analysis using the data obtained in all approaches used in this study resulted in 22 groups (16 groups among the 24 high-toxicity strains) at 100% similarity, indicating a high degree of diversity among the strains tested. Some of the strains studied here, which are resistant to different stress conditions, should be considered for further ecological studies.