Bovine mastitis is a disease with a high economic impact on dairy farms and it has been described that different species of Candida can cause it. The objectives of this study were to evidence the production of biofilm by strains of Candida Kefyr, to carry out a comparative evaluation of its production by tube and plate techniques, to determine its interaction with Escherichia coli and Streptococcus dysgalactiae isolated from bovine mastitis and know the drug susceptibility of each of them. The identification of C. kefyr was carried out by fermentation and assimilation of carbohydrates, the confirmation of the identification and minimum inhibitory concentration to antimycotics was carried out with Vitek 2 Systems. Plate and tube biofilm formation assays were performed in triplicate. Analysis of variance (ANOVA) and Tukey's test were performed, as well as Spearman's correlation to the data obtained. In plate tests, biofilm formation was demonstrated in seven C. kefyr isolates, as well as synergy in combinations with E. coli and S. dysgalactiae. However, the inoculum containing the three microorganisms behaved similarly to that containing only C. kefyr and S. dysgalactiae. It was possible to demonstrate a correlation between biofilm formation tests in tube and plate. This is the first report in Mexico of C. kefyr, as well as the production of biofilm in bovine clinical mastitis.