This study was conducted to examine effects of the dose and viability of supplemental Saccharomyces cerevisiae on the ruminal fermentation and bacteria population and the performance of lactating dairy cows. Four ruminally cannulated lactating cows averaging 284±18d in milk were assigned to 4 treatments arranged in a 4×4 Latin square design with four 21-d periods. Cows were fed a total mixed ration containing 41.7% corn silage, 12.1% brewer's grains, and 46.2% concentrate on a dry matter basis. The diet was supplemented with no yeast (control) or with a low dose of live yeast (5.7×10 cfu/cow per day; LLY), a high dose of live yeast (6.0×10 cfu/cow per day; HLY), or a high dose of killed yeast (6.0×10 cfu/cow per day; HDY). Microbial diversity was examined by high-throughput Illumina MiSeq sequencing (Illumina Inc., San Diego, CA) of the V4 region of the 16S rRNA gene. The relative abundance of select ruminal bacteria was also quantified by quantitative PCR (qPCR). Adding LLY to the diet increased the relative abundance of some ruminal cellulolytic bacteria (Ruminococcus and Fibrobacter succinogenes) and amylolytic bacteria (Ruminobacter, Bifidobacterium, and Selenomonas ruminantium). Adding live instead of killed yeast increased the relative abundance of Ruminococcus and F. succinogenes; adding HDY increased the relative abundance of Ruminobacter, Bifidobacterium, Streptococcus bovis, and Selenomonas ruminantium. The most dominant (≥1% of total sequences) bacteria that responded to LLY addition whose functions are among the least understood in relation to the mode of action of yeast include Paraprevotellaceae, CF231, Treponema, and Lachnospiraceae. Future studies should aim to speciate, culture, and examine the function of these bacteria to better understand their roles in the mode of action of yeast. A relatively precise relationship was detected between the relative abundance of F. succinogenes (R=0.67) from qPCR and MiSeq sequencing, but weak relationships were detected for Megasphaera elsdenii, Ruminococcus flavefaciens, and S. ruminantium (R≤0.19).

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Previously we investigated the impact of a mixed Lactobacillus buchneri, Lactobacillus plantarum, Lactobacillus casei inoculant on fermentation and aerobic stability of barley silage over two years in 2009 and 2010. In 2009, a classical response to inoculation was obtained with an increase in acetic acid concentration of silage ensiled in both mini- and bag silos. In 2010, this classical response was not observed in mini-silos, but was observed in bag silos. The objective of this study was to determine if molecular microbial ecology could explain the differential responses to the inoculation of barley silage between the two years. The Illumina MiSeq sequencing results showed that inoculation increased Lactobacillus and lowered Pediococcus, Weissella and Leuconostoc in both types of silos in 2009. However, a similar trend was not observed in mini-silos but was observed in bag silos in 2010. Inoculation did not alter the core fungal community in either silo type in either year. Cladosporium, Leptosphaeria and Cryptococcus were abundant in fresh forage, but were superseded by Pichia and Kazachstania after ensiling. Our results suggest that changes in silage chemistry corresponded to differences observed in microbial ecology. Inoculation may have less impact when using more mature crops with shorter ensiling times.

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