Microbial diversity and stability during primary cultivation and subcultivation processes of Tibetan kefir

Summary In this study, we have investigated the microbial diversity and stability in the Tibetan kefir during the primary cultivation and subcultivation processes via a combination of culture-independent and culture-dependent methods. According to the culture-independent methods, the profiles of PCR-DGGE (polymerase chain reaction–denaturing gradient gel electrophoresis) indicated that nine microbial species were predominant at different cultivation stages; seventy four isolates of seven predominant species were obtained and identified via the culture-dependent methods. PCR-DGGE further confirmed that Lactobacillus kefiranofaciens, Lactobacillus kefiri, Lactobacillus paracasei, Streptococcus thermophilus, Kazachstania unispora and Saccharomyces cerevisiae were the most dominant species in the Tibetan kefir during the process of primary cultivation, among which Lb. kefiri, Lb. paracasei and K. unispora showed relative strong stability during both the processes of primary cultivation and subcultivation. These findings suggested that some isolates of the three species possessed the potentiality of being used in the development of direct vat set (DVS) starters for the production of Tibetan kefir and the related products.