| Summary: | This study assessed the effects of <i>Lactiplantibacillus plantarum</i> (ZZU203), cellulase-producing <i>Bacillus methylotrophicus</i> (CB), or their combination (ZZU203_CB) on the fermentation parameters of alfalfa after 10 and 60 days of ensiling. Additionally, the bacterial community compositions were analyzed using absolute quantification 16S-seq (AQS). The results showed that CB silage displayed a higher lactic acid (LA) concentration at 10 d, a higher abundance of <i>Lactobacillus</i>, and lower abundance of <i>Pediococcus</i>, <i>Enterococcus</i>, and <i>Weissella</i> than those in the control (CK) silage. Compared with CK silage, the ZZU203 silage increased LA concentration, fructose and rhamnose concentrations, and the abundance of <i>Lactobacillus</i>, and decreased pH value, ammoniacal nitrogen, acetic acid, neutral detergent fiber and acid detergent fiber concentrations, and the abundance of <i>Pediococcus</i>, <i>Enterococcus</i>, <i>Weissella</i>, <i>Hafnia</i>, and <i>Garciella</i> after 60 days of ensiling. In addition, ZZU203 and ZZU203_CB silage had a similar silage quality and bacterial community, while the inoculation of ZZU203_CB significantly promoted LA accumulation and the numbers of <i>Lactobacillus</i> at 10 d compared with ZZU203 silage. Therefore, ZZU203 or a combination of ZZU203 and CB can be used as potential silage additives to improve the silage quality of alfalfa.
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