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Endospore-forming bacteria related to the  Bacillus cereus  group produce toxins that cause illnesses in organisms from invertebrates to mammals, including foodborne illnesses in humans. As commercial bee pollen can be contaminated with these bacteria, a comprehensive microbiological risk assessment of commercial bee pollen must be incorporated into the relevant regulatory requirements, including those that apply in Mexico. To facilitate detection of members of this group of bacteria, we have developed a PCR strategy that is based on the amplification of the single-copy tRNA Cys  gene and specific genes associated with tRNA Cys  to detect  Bacillus cereus sensu lato  ( B. cereus s.l. ). This tRNA Cys -PCR-based approach was used to examine commercial bee pollen for endospore-forming bacteria. Our analysis revealed that 3% of the endospore-forming colonies isolated from a commercial source of bee pollen were related to  B. cereus s.l.,  and this result was corroborated by phylogenetic analysis, bacterial identification via MALDI-TOF MS, and detection of enterotoxin genes encoding the HBL and NHE complexes. The results show that the isolated colonies are closely related phylogenetically to  B. cereus, B. thuringiensis , and  B. bombysepticus . Our results indicate that the tRNA Cys -PCR, combined with other molecular tools, will be a useful approach for identifying  B. cereus s.l.  and will assist in controlling the spread of potential pathogens.

Phylogenetic Analysis of Bacillus cereus sensu lato Isolates from Commercial Bee Pollen Using tRNACys-PCR