Effect of Isolation Ruminal Yeast from Ruminants on In Vitro Ruminal Fermentation.

Abstract

In order to obtain high-performing yeast strains from ruminants, it is necessary to select them from species such as beef cattle, dairy cows, goats, and buffalo. A total of 91 isolated yeasts were collected using the standard methods of microbial culture on agar medium followed by streaking on a plate at least three times until pure yeast colonies were formed. The API 20C AUX Kit and sequencing of the D1/D2 domain of the 26S rRNA gene were used to identify the genera Candida spp., namely, C. glabrata (99% identification), C. tropicallis (99%), C. rugosa (98%), and Issatchenkia orientalis (99%). A total of 12 yeast strains (Dc4, 14, 18; Be1, 2, 7; Bu3, 4, 7; and Go10, 16, 19) were chosen for further analyses. The performance criteria included the ability to tolerate pH values between 3.5 and 7.5, total volatile fatty acids (TVFAs, 0, 0.25, 0.5, 1, 2, and 4% of broth medium), anaerobic growth rate, and in vitro gas production efficiency. First, when all strains were grown at pH values between 3.5 and 7.5, Bu3 and Dc18 performed better than the other strains. Second, at a ruminal pH of 6.5 and a TVFA concentration of between 2 and 4% of the broth medium, strain Bu3 was more resistant than the other strains. Under anaerobic conditions, all strains experienced a decline in viable cell counts when compared with those under aerobic conditions. However, compared to strains Dc14, Be1, Be2, Be7, and Bu3, strain Dc18 exhibited more viable cells under anaerobic conditions in broth medium. The response of strain Dc18 did not differ from those of strains Dc4, Bu4, Bu7, or G16. Strains Be7, Bu3, and Dc18 were used for an in vitro fermentation experiment involving incubation for 2, 4, 6, 8, 10, 12, 24, 36, 48, and 72 h. Three ruminal cannulated dairy cows were used as donors of ruminal fluid. The treatments were run in triplicate. The addition of yeast culture had no effect on gas kinetics, gas accumulation, or the ratio of acetic acid and propionic acid, but led to significantly greater butyric acid concentrations at 24 h of incubation. In conclusion, strain Dc18 isolated from dairy cows is suitable for future studies of probiotic yeast development.