Effects of Moisture Content Gradient on Alfalfa Silage Quality, Odor, and Bacterial Community Revealed by Electronic...

Better quality and odor of silage and normal microbial fermentation metabolism are mostly dependent on an appropriate moisture content. The purpose of this study was to determine the effects of different moisture content gradients (50, 60, 70, and 80%) on the bacterial community, odor, and quality of alfalfa silage at 60 days by using gas chromatography-mass spectrometry (GC-MS) and electronic nose, with six replicates per group. The results showed that there were significant differences in odor response intensity among all groups, among which the 80% group had the strongest reaction to terpenoids, sulfides, and nitrogen oxides. Similarly, the different volatile organic compounds (VOCs) were mainly terpenoids, alcohols, and ketones, such as pine, camphor, and menthol (e.g., carlin and levomenthol). The dominant bacterium was Enterococcus with higher fiber, pH, and ammonia nitrogen (NH₃-N) content but poorer quality and odor (p < 0.05). The differential VOCs in the 60% group were mainly heterocyclics, esters, and phenols with fruity, floral, and sweet odors such as 2-butylthiophene and acorone. Pediococcus and Lactiplantibacillus were the dominant bacteria, with higher crude protein (CP), water-soluble carbohydrates (WSC), and lactic acid (LA) contents, as well as better quality and odor (p < 0.05). The biosynthesis of terpenoids and steroids, biosynthesis of secondary metabolites, and biosynthesis of phenylpropanoids were the main metabolic pathways of differential VOCs. In conclusion, regulating moisture content can alter bacterial community and metabolites, which will encourage fermentation and enhance alfalfa silage quality and odor.