Background and Aim: Probiotic supplementation can assist with manipulating the rumen microbial ecosystem. Lactic acid bacteria and yeast from fermented fish (Budu) as the indigenous food from West Sumatra, Indonesia, are potential probiotics for livestock. This study aims to select the best candidate lactic acid bacteria and yeast strains from fermented fish as ruminant probiotics and evaluate the effect of their supplementation on the characteristics of rumen fermentation, feed digestion, and total gas production in vitro.

Materials and Methods: This study used nine treatments, performed in triplicate, in a completely randomized design. The substrate ratio comprised of 70% Pennisetum purpureum forage and 30% concentrate. Five lactic acid bacteria and three yeast isolates were used in this study. Treatments were as follows: T0: control (basal diet); T1: T0 + Lactobacillus parabuchneri strain 3347; T2: T0 + Lactobacillus buchneri strain 5296; T3: T0 + Lactobacillus harbinensis JCM 16178; T4: T0 + Schleiferilactobacillus harbinensis strain LH991; T5: T0 + L. parabuchneri strain 6902; T6: T0 + Pichia kudriavzevii strain B-5P; T7: T0 + P. kudriavzevii strain CBS 5147; and T8: T0 + commercial yeast (Saccharomyces cerevisiae). The lactic acid bacteria inoculum contained 1.02 × 10¹¹ colony-forming unit (CFU)/mL, while the yeast inoculum contained 1.5 × 10¹⁰ CFU/mL.

Results: The results showed that four lactic acid bacteria and three yeast produced a higher total gas yield (104–183.33 mL) compared to the control (103 mL). Supplementation with lactic acid bacteria in the rumen fermentation in vitro showed dry matter digestibility of 63%–70% and organic matter digestibility (OMD) of 64%–71%. We observed that total volatile fatty acid (VFA) production in all treatments was significantly higher (86–121 mM) compared to the control (81 mM). The concentration of NH₃ production was higher in all treatments (12.33–16.83 mM) than in the control (12.25 mM). Meanwhile, the probiotic supplementation did not cause a significant change in the rumen pH (6.86–7.12). Supplementation with the lactic acid bacteria S. harbinensis strain LH991 consistently demonstrated the best results from the parameters of dry and OMD (70.29% and 71.16%, respectively), total VFA (121.67 mM), NH₃ (16.83 mM), and total gas production (149.17 mL). The best results were observed from the yeast candidate P. kudriavzevii strain B-5P, where the results were dry and OMD (67.64% and 69.55% respectively), total VFA (96.67 mM), NH₃ (13.42 mM), and total gas production (183.33 mL).

Conclusion: Based on the obtained results, lactic acid bacteria S. harbinensis strain LH991 and yeast P. kudriavzevii strain B-5P are attractive candidates to be utilized as probiotics for ruminants based on their potential to improve rumen fermentation in vitro. This probiotic supplementation can increase the digestibility of feed ingredients, production of total VFA and NH₃, and total gas produced.

Keywords: in vitro, lactic acid bacteria, probiotics, rumen fermentation, yeast.