Thirty-three strains of Brevibacillus laterosporus, including three novel strains isolated from Brazilian soil samples, were examined for genetic variability by the use of different PCR-based methods. Molecular markers that could characterize bacterial strains with regards to their pathogenic potential were investigated. In addition, toxicity was assessed by the use of insects belonging to the orders Lepidoptera and Coleoptera and the mollusk Biomphalaria glabrata. Among the targets tested, Biomphalaria glabrata demonstrated the highest degree of sensitivity to B. laterosporus, with some strains inducing 90 to 100% mortality in snails aged 3 and 12 days posteclosion. Larvae of the coleopteron Anthonomus grandis were also susceptible, presenting mortality levels of between 33 and 63%. Toxicity was also noted towards the lepidopteron Anticarsia gemmatalis. In contrast, no mortality was recorded among test populations of Tenebrio molitor or Spodoptera frugiperda. The application of intergenic transcribed spacer PCR and BOX-PCR generated 15 and 17 different genotypes, respectively. None of the molecular techniques allowed the identification of a convenient marker that was associated with any entomopathogenic phenotype. However, a 1,078-bp amplicon was detected for all strains of B. laterosporus when a primer for amplification of the BOXA1R region was used. Similarly, a 900-bp amplicon was generated from all isolates by use of the primer OPA-11 for randomly amplified polymorphic DNA analysis. These amplicons were not detected for other phenotypically related Brevibacillus species, indicating that they represent markers that are specific for B. laterosporus, which may prove useful for the isolation and identification of new strains of this species.

The aerobic endospore-forming bacteria (AEFB) comprise species of Bacillus and related genera and have long been regarded as prominent constituents of the soil bacterial community. The wide diversity of AEFB renders appropriate categorisation and generalisations a challenging task. We previously isolated 312 AEFB strains from Brazilian soils that we designated SDF (Solo do Distrito Federal) strains. To better understand the SDF diversity and explore their biotechnological potential, we addressed the biochemical and physiological profiles of these 312 environmental strains by performing 30 tests in this work. Of these, the 16S rRNA gene sequences segregated 238 SDF strains into four genera in the family Bacillaceae and two in the Paenibacillaceae. Bacillus spp. were the most prevalent, followed by species of Paenibacillus. We summarised the phenotypic test relationships among selected SDF strains using a Pearson correlation-based clustering represented in heatmaps. In practice, biochemical and physiological profiles are often less discriminatory than molecular data and may be unstable because of the loss of traits. Although these test reactions are not universally positive or negative within species, they may define biotypes and be efficient strain markers, enhancing the accuracy of unknown sample identification. It can also help select the most representative phenotypes of samples. Along with the other phenotypic and genotypic data, the present results are of great importance for the robust classification of the SDF strains within the scope of the polyphasic approach.