Abstract The objective was to characterize the microflora of Mexican artisanal Fresco cheese, as a first step towards the development of a starter culture, which would allow the making of a safe and uniform product from pasteurized milk. The numbers of several microbial groups (aerobic mesophilic flora, total and fecal coliforms, Salmonella spp., Staphylococcus spp., molds, yeasts and lactic acid bacteria) were investigated. The dominating isolates were characterized as being Lactococcus lactis ssp. lactis, Enterococcus faecium and Lactobacillus casei. The results suggest that an adequate starter in the production of Mexican Fresco cheese may be composed of strains of these species.