Medycyna Wet. 67 (8), 545-549, 2011
Krutkiewicz A., Szopa D., Kleczkowski M., Biegańska M., Dworecka-Kaszak B.
Restriction analysis of Candida strains isolated from milk samples obtained from cows with clinical mastitis
The aim of the study was to obtain genotypic differentiation of fungi isolated from milk samples of dairy cows showing clinical signs of mastitis. Twenty strains of fungi were isolated from milk samples and identified as Candida, and then classified into seven different species by API Candida and API ID 32C tests (bioMerieux). Next, the genomic DNA was isolated from each fungal strain and amplified with ITS1 and NL2 primers. Amplification products were digested with HpaII and EcoRI restriction enzymes, while the restriction profiles were resolved in VersaDoc imaging apparatus with the assistance of Quantity One software. PCR with ITS1 and NL2 primers produced DNA fragments of various lengths, raging from 700 to 1000 bp. Their molecular weight was dependent on the fungal species from which DNA was isolated. Comparison of restriction fragment length confirmed differences between species; however, strains similarly classified based on phenotypic characteristics also revealed differences in the restriction fragment profile. For none of the investigated species was a characteristic, uniform genetic profile obtained. Results of the presented study revealed that the examined species did not give a uniform genetic profile, suggesting that Candida strains phenotypically belonging to the same species may have varied genotypes in the analyzed restrictive places.
Key words: mastitis, Candida, restriction analyses, genotypic methods