Effects of inoculating feruloyl esterase-producing Lactiplantibacillus plantarum A1 on ensiling characteristics,in vitro ruminal fermentation and microbiota of alfalfa silage

Background Ferulic acid esterase(FAE)-secreting Lactiplantibacillus plantarum A1(Lp A1)is a promising silage inoculant due to the FAE’s ability to alter the plant cell wall structure during ensiling,an action that is expected to improve forage digestibility.However,little is known regarding the impacts of Lp A1 on rumen microbiota.Our research assessed the influences of Lp A1 in comparison to a widely adopted commercial inoculant Lp MTD/1 on alfalfa’s ensilage,in vitro rumen incubation and microbiota.Results Samples of fresh and ensiled alfalfa treated with(either Lp A1 or Lp MTD/1)or without additives(as control;CON)and ensiled for 30,60 and 90 d were used for fermentation quality,in vitro digestibility and batch culture study.Inoculants treated silage had lower(P<0.001)pH,acetic acid concentration and dry matter(DM)loss,but higher(P=0.001)lactic acid concentration than the CON during ensiling.Compared to the CON and Lp MTD/1,silage treated with Lp A1 had lower(P<0.001)aNDF,ADF,ADL,hemicellulose,and cellulose contents and higher(P<0.001)free ferulic acid concentration.Compared silage treated with Lp MTD/1,silage treated with Lp A1 had significantly(P<0.01)improved ruminal gas production and digestibility,which were equivalent to those of fresh alfalfa.Realtime PCR analysis indicated that Lp A1 inoculation improved the relative abundances of rumen’s total bacteria,fungi,Ruminococcus albus and Ruminococcus flavefaciens,while the relative abundance of methanogens was reduced by Lp MTD/1 compared with CON.Principal component analysis of rumen bacterial 16S rRNA gene amplicons showed a clear distinction between CON and inoculated treatments without noticeable distinction between Lp A1 and Lp MTD/1 treatments.Comparison analysis revealed differences in the relative abundance of some bacteria in different taxa between Lp A1 and Lp MTD/1 treatments.Silage treated with Lp A1 exhibited improved rumen fermentation characteristics due to the inoculant effects on the rumen microbial populations and bacterial community.Conclusions Our findings suggest that silage inoculation of the FAE-producing Lp A1 could be effective in improving silage quality and digestibility,and modulating the rumen fermentation to improve feed utilization.