Background:Bacillus cereusis associated with milk, dairy product, and dairy farm contamination. The aim of this study was to characterize strains ofB. cereusin the small-scale artisanal cheese production chain in southwestern Mexico. Methods: 130 samples were collected.B. cereusisolation was performed on Mannitol Egg Yolk Polymyxin (MYP) agar. Genotyping, enterotoxigenic profile, and determination of genes involved in the formation ofB. cereusbiofilm were performed by PCR. An antimicrobial susceptibility test was made by broth microdilution assay. The phylogenetic analysis was performed by amplification and sequencing of 16s rRNA. Results:B. cereus sensu latowas isolated and molecularly identified in 16 samples andB. cereus sensu stricto(B. cereus) was the most frequently isolated and identified species (81.25%). Of all the isolatedB. cereus sensu latostrains, 93.75% presented at least one gene for some diarrheagenic toxins, 87.5% formed biofilms, and 18.75% were amylolytic. AllB. cereus sensu latostrains were resistant to beta-lactams and folate inhibitors. A close phylogenetic relationship between isolates was found between the cheese isolates and the air isolates. Conclusions: Strains ofB. cereus sensu latowere found in small-scale artisanal cheeses on a farm in southwestern Mexico.