Insights into microbiota community dynamics and flavor development mechanism during golden pomfret(Trachinotus ovatus)fermentation based on single-molecule real-time sequencing and molecular networking analysis

Popular fermented golden pomfret(Trachinotus ovatus)is prepared via spontaneous fermentation;however,the mechanisms underlying the regulation of its flavor development remain unclear.This study shows the roles of the complex microbiota and the dynamic changes in microbial community and flavor compounds during fish fermentation.Single-molecule real-time sequencing and molecular networking analysis revealed the correlations among different microbial genera and the relationships between microbial taxa and volatile compounds.Mechanisms underlying flavor development were also elucidated via KEGG based functional annotations.Clostridium,Shewanella,and Staphylococcus were the dominant microbial genera.Forty-nine volatile compounds were detected in the fermented fish samples,with thirteen identified as characteristic volatile compounds(ROAV>1).Volatile profiles resulted from the interactions among the microorganisms and derived enzymes,with the main metabolic pathways being amino acid biosynthesis/metabolism,carbon metabolism,and glycolysis/gluconeogenesis.This study demonstrated the approaches for distinguishing key microbiota associated with volatile compounds and monitoring the industrial production of high-quality fermented fish products.

Structure and immunomodulatory activity of Lentinus edodes polysaccharides modified by probiotic fermentation

Plant-based fermentations provide an untapped source for novel biotechnological applications.In this study,a probiotic named Lactobacillus fermentum 21828 was introduced to ferment Lentinus edodes.Polysaccharides were extracted from fermented and non-fermented L.edodes and purified via DEAE-52 and Sephadex G-100.The components designated F-LEP-2a and NF-LEP-2a were analyzed by FT-IR,HPGPC,HPAEC,SEM,GC-MS and NMR.The results revealed that probiotic fermentation increased the molecular weight from 1.16×10^(4) Da to 1.87×10^(4) Da and altered the proportions of glucose,galactose and mannose,in which glucose increased from 45.94%to 48.16%.Methylation analysis and NMR spectra indicated that F-LEP-2a and NF-LEP-2a had similar linkage patterns.Furthermore,their immunomodulatory activities were evaluated with immunosuppressive mice.NF-LEP and F-LEP improved immune organ indices,immunoglobulin(Ig G and Ig M)and cytokines concentrations;restored the antioxidation capacity of liver;and maintained the balance of gut microbiota.F-LEP displayed better moderating effects on the spleen index,immunoglobulin,cytokines and the diversity of gut microbiota than NF-LEP(200,400 mg/kg).Our study provides an efficient and environment-friendly way for the structural modification of polysaccharides,which helps to enhance their biological activity and promote their wide application in food,medicine and other fields.

Effects of Rosa roxburghii&edible fungus fermentation broth on immune response and gut microbiota in immunosuppressed mice

With the rise of probiotics fermentation in food industry,fermented foods have attracted worldwide attention.In this study,protective effects of Rosa roxburghii&edible fungus fermentation broth(REFB)on immune function and gut health in Cyclophosphamide induced immunosuppressed mice were investigated.Results showed that REFB could improve the immune organ index,and promote the proliferation and differentiation of splenic T lymphocytes.In addition,it attenuated intestinal mucosal damage and improved intestinal cellular immunity.REFB administration also up-regulated the expression of IL-4,INF-γ,TNF-α,T-bet and GATA-3 mRNA in small intestine.Furthermore,administration of REFB modulated gut microbiota composition and increased the relative abundance of beneficial genus,such as Bacteroides.It also increased the production of fecal short-chain fatty acids.These indicate that REFB has the potential to improve immunity,alleviate intestinal injury and regulate gut microbiota in immunosuppressed mice.

The Advantages of Methane Production by Combined Fermentation of Lignite and Wheat Straw

Biogasification of coal is important for clean utilization of coal. Experiments on the fermentation of single lignite, single straw and their mixture were performed to explore the variation characteristics of gas production potential, microbial community and methanogenic metabolic pathways of mixture. Research has shown that mixed fermentation of lignite and straw significantly promoted biomethane production. The abundance of hydrolytic acidifying functional bacteria genera (Sphaerochaeta, Lentimicrobium) in mixed fermentation was higher than that in the fermentation of single lignite and single straw. The abundance of some key CAZy metabolic enzyme gene sequences in mixed fermentation group was increased, which was favorable to improve methane production. Aceticlastic methanogenesis was the most critical methanogenic pathway and acetic acid pathway was more competitive in methanogenic mode during peak fermentation. Macrogenomics provided theoretical support for the claim that mixed fermentation of coal and straw promoted biomethane metabolism, which was potentially valuable in expanding methanogenesis from mixed fermentation of lignite with different biomasses.

Postbiotics from Saccharomyces cerevisiae fermentation stabilize microbiota in rumen liquid digesta during grain-based subacute ruminal acidosis(SARA) in lactating dairy cows

Background Subacute ruminal acidosis(SARA)is a common metabolic disorder of high yielding dairy cows,and it is associated with dysbiosis of the rumen and gut microbiome and host inflammation.This study evaluated the impact of two postbiotics from Saccharomyces cerevisiae fermentation products(SCFP)on rumen liquid associated microbiota of lactating dairy cows subjected to repeated grain-based SARA challenges.A total of 32 rumen cannulated cows were randomly assigned to 4 treatments from 4 weeks before until 12 weeks after parturition.Treatment groups included a Control diet or diets supplemented with postbiotics(SCFPa,14 g/d Original XPC;SCFPb-1X,19 g/d Nutri Tek;SCFPb-2X,38 g/d Nutri Tek,Diamond V,Cedar Rapids,IA,USA).Grain-based SARA challenges were conducted during week 5(SARA1)and week 8(SARA2)after parturition by replacing 20%DM of the base total mixed ration(TMR)with pellets containing 50%ground barley and 50%ground wheat.Total DNA from rumen liquid samples was subjected to V3–V416S r RNA gene amplicon sequencing.Characteristics of rumen microbiota were compared among treatments and SARA stages.Results Both SARA challenges reduced the diversity and richness of rumen liquid microbiota,altered the overall composition(β-diversity),and its predicted functionality including carbohydrates and amino acids metabolic pathways.The SARA challenges also reduced the number of significant associations among different taxa,number of hub taxa and their composition in the microbial co-occurrence networks.Supplementation with SCFP postbiotics,in particular SCFPb-2X,enhanced the robustness of the rumen microbiota.The SCFP supplemented cows had less fluctuation in relative abundances of community members when exposed to SARA challenges.The SCFP supplementation promoted the populations of lactate utilizing and fibrolytic bacteria,including members of Ruminococcaceae and Lachnospiraceae,and also increased the numbers of hub taxa during non-SARA and SARA stages.Supplementation with SCFPb-2X prevented the fluctuations in the abundances of hub taxa that were positively correlated with the acetate concentration,andα-andβ-diversity metrics in rumen liquid digesta.Conclusions Induction of SARA challenges reduced microbiota richness and diversity and caused fluctuations in major bacterial phyla in rumen liquid microbiota in lactating dairy cows.Supplementation of SCFP postbiotics could attenuate adverse effects of SARA on rumen liquid microbiota.

Influence of nitrogen status on fermentation performances of non-Saccharomyces yeasts:a review

Nitrogen,one of the most crucial nutrients present in grapes and musts,plays a key role in yeast activities during alcoholic fermentation.Such influences are imposed on yeast growth and fermentation performances including the formation of secondary metabolites.Saccharomyces cerevisiae,the main yeast responsible for fermentation,has been studied extensively regarding nitrogen impacts.On the other hand,a similar study for non-Saccharomyces yeasts,whose contributions to winemaking have gradually been acknowledged,remains to be fully explored,with a few studies being reported.This review starts by discussing nitrogen impacts on non-Saccharomyces yeast growth and fermentation kinetics in different case scenarios,then proceeds to summarize the nitrogen preferences of individual yeast strains with regulation mechanisms elucidated by recent studies.Detailed discussions on the influences on the production of volatile compounds and proposed pathways therein are made,followed by future work suggested as the final section.In summarizing the nitrogen impacts on non-Saccharomyces yeasts throughout alcoholic fermentation,this review will be helpful in obtaining a more comprehensive view on these non-conventional wine yeasts in terms of nutrient requirements and corresponding volatile production.Research gaps will therefore be elucidated for future research.

The convergence of lactic acid microbiomes and metabolites in long-term electrofermentation

Regulating electron transfer in predominantly fermentative microbiomes has broad implications in environmental,chemical,food,and medical fields.Here we demonstrate electrochemical control in fermenting food waste,digestate,and wastewater to improve lactic acid production.We hypothesize that applying anodic potential will expedite and direct fermentation towards lactic acid.Continued operation that introduced epi/endophytic communities(Lactococcus,Lactobacillus,Weissella)to pure culture Lactiplantibacillus plantarum reactors with static electrodes was associated with the loss of anode-induced process intensification despite 80%L.plantarum retention.Employing fluidized electrodes discouraged biofilm formation and extended electrode influence to planktonic gram-positive fermenters using mediated extracellular electron transfer.While short-term experiments differentially enriched Lactococcus and Klebsiella spp.,longer-term operations indicated convergent microbiomes and product spectra.These results highlight a functional resilience of environmental fermentative microbiomes to perturbations in redox potential,underscoring the need to better understand electrode induced polymicrobial interactions and physiological impacts to engineer tunable open-culture or synthetic consortia.

The Influence of Gratering, Fermentation and Variety on the Physio-Chemical Quality of Cassava Starch

The study was conducted to determine the influence of gratering and fermentation parameters on the physicochemical quality of starch obtained from two cassava varieties in Sierra Leone (i.e., SLICASS 11 and SLICASS 6). Fresh cassava roots harvested from the Department of Agricultural Engineering were peeled, washed and grated before fermentation and starch extraction. Fermentation was carried out under separate aerobic and anaerobic conditions for a period of Seven days. Physicochemical analyses were conducted at the Postharvest Food and Bioprocess Engineering Laboratory of the Department of Agricultural and Bio-System Engineering, Njala University to determine the solubility, water absorption capacity and swelling power of starch extracts obtained from various experimental treatments. Fermentation method and duration had significant effects on the solubility, water absorption capacity (WAC) and swelling power (SP) of starch extracts. Maximum solubility and WAC were recorded on the fifth day, for both cassava varieties tested, with apparent significant difference resulting from the two gratering bands (i.e., with 1.5-inch nail hole and 4-inch nail hole sizes, respectively). While swelling power increased consistently with temperature for starch obtained from SLICASS-11 variety, an irregular pattern was observed for SLICASS-6 variety. A multiple correlation analysis proposes a significant and weak correlation between temperature, WAC (+0.150) and swelling power (+0.048). Also multiple correlation analyses suggest a significant correlation between fermentation period, the functional properties of starch extracts obtained from both fermentation methods and cassava varieties (i.e., solubility (−0.226), water absorption capacity (+0.301) and swelling power (+0.329)).

Microbial metabolic interaction in fermentation ecosystem and cooperation in flavor compounds formation of Chinese cereal vinegar

Shanxi aged vinegar(SAV)is a famous cereal vinegar in China,which is produced through a solid-state fermentation where diverse microbes spontaneously and complex interactions occur.Here,combined with the metatranscriptomics,the microbial co-occurrence network was constructed,indicating that Lactobacillus,Acetobacter and Pediococcus are the most critical genera to maintain the fermentation stability.Based on an extensive collection of 264 relevant literatures,a transport network containing 2271 reactions between microorganisms and compounds was constructed,showing that glucose(84%of all species),fructose(67%)and maltose(67%)are the most frequently utilized substrates while lactic acid(64%),acetic acid(45%)are the most frequently occurring metabolites.Specifically,the metabolic influence of species pairs was calculated using a mathematical calculation model and the metabolic influence network was constructed.The topology properties analysis found that Lactobacillus was the key role with robust metabolic control of vinegar fermentation ecosystem and acetic acid and lactic acid were the main metabolites with feedback regulation in microbial metabolism of SAV.Furthermore,systematic coordination of positive and negative impacts was proved to be inevitable to form flavor compounds and maintain a natural microbial ecosystem.This study provides a new perspective for understanding microbial interactions in fermented food.

Impact of sourdough fermentation on nutrient transformations in cereal-based foods:Mechanisms,practical applications,and health implications

Sourdough is often considered a healthy choice and quality improver for use in cereal production due to its unique microbial composition and fermentation properties.During sourdough fermentation of cereals,biotransformation of nutrients occurs,resulting in notable changes to proteins,carbohydrates,fats,vitamins,and minerals.Each nutrient undergoes specific transformations,providing various advantages for human health.Proteins undergo hydrolysis to produce small molecular weight peptides and amino acids that are more easily digested and absorbed by the human body.Carbohydrates break down to improve the digestibility and absorption of cereals and lower the glycemic index.Fatty acids experience oxidation to produce new substances with health benefits.Additionally,the application of sourdough fermentation can enhance the texture,flavor,and nutritional value of cereal foods while also extending their shelf life and improving food safety.In conclusion,sourdough fermentation has a broad range of applications in cereal food processing.Further research is encouraged to investigate the mechanisms and processes of sourdough fermentation to develop even more nutritious,healthy,and flavorful cereal-based foods.

The influence of Lactobacillus plantarum fermentation in selenium-enriched Brassica napus L.:changes in the nutritional constituents,bioactivities and bioaccessibility

Selenium(Se)-enriched Brassica napus L.is a valuable organic Se supplement.In this study,the fermentation broth enriched with organic Se(FFS)was prepared using Lactobacillus plantarum to ferment the substrate of Se-enriched Brassica napus L.Significant increases were observed after fermentation in total sugars,reducing sugars,soluble proteins,total phenolic content(TPC),and total flavonoid content(TFC).The organic Se was retained at a concentration of 54.75 mg/g in the freeze-dried sample.Principal component analysis and cluster analysis showed good separation between the FFS and unfermented(FS)groups.Fragrant 2-ethyloxetane had the highest content among all volatiles,while sinapine had the highest content among all phenolic compounds.The fermentation process showed remarkable improvement in the abundance and concentration of volatile compounds and phenolic contents,making FFS exhibit strong antioxidant activity and inhibitory capacity againstα-glucosidase activity.The bioaccessibility of phenolic compounds was significantly greater in FFS compared to FS.ADMET analysis revealed that the majority of phenolic compounds contained in FFS did not exhibit mutagenicity toxicity,hepatotoxicity,skin sensitization,or blood-brain barrier penetration,indicating a favorable level of biosafety.Overall,our study provides a new insight into the further utilization of Se-enriched Brassica napus L.in foods.

Screening of glucosinolates degrading lactic acid bacteria and their utilization in rapeseed meal fermentation

Rapeseed meal is a promising food ingredient, but its utilization is limited by the presence of some potentially harmful ingredients, such as glucosinolates. Fermentation is a cost-effective method of detoxication but a food-grade starter culture with glucosinolates degradation capacity is required. In this study, 46 strains of lactic acid bacteria from traditional paocai brines were screened for their ability to glucosinolate degradation. The results showed that more than 50% of the strains significantly degraded glucosinolates. Two strains of Lactiplantibacillus(p7 and s7) with high capacity of glucosinolates degradation through producing enzymes were identified. Then,an optimized condition for rapeseed meal fermentation by p7 was established to degrade glucosinolates, which can achieve about 80% degradation. UPLC/Q-TOF-MS analysis showed that the degradation rate of individual glucosinolates was different and the degradation rate of gluconapin and progoitrin in rapeseed meal can reach more than 90%. Meanwhile, fermentation with p7 can improve safety of rapeseed meal by inhibiting the growth of Enterobacteriaceae and improve its nutritional properties by degrading phytic acid. The in vitro digestion experiments showed that the content of glucosinolates in rapeseed meal decreased significantly during gastric digestion. Meanwhile, fermentation with p7 can greatly improve the release of soluble protein and increase the contents of free essential amino acids, such as lysine(increased by 12 folds) and methionine(increased by 10 folds).

Allergen degradation of bee pollen by lactic acid bacteria fermentation and its alleviatory effects on allergic reactions in BALB/c mice

Food allergy as a global health problem threatens food industry.Bee pollen(BP)is a typical food with allergenic potentials,although it performs various nutritional/pharmacological functions to humans.In this study,lactic acid bacteria(LAB)were used to ferment Brassica napus BP for alleviating its allergenicity.Four novel allergens(glutaredoxin,oleosin-B2,catalase and lipase)were identified with significant decreases in LAB-fermented BP(FBP)than natural BP by proteomics.Meanwhile,metabolomics analysis showed significant increases of 28 characteristic oligopeptides and amino acids in FBP versus BP,indicating the degradation of LAB on allergens.Moreover,FBP showed alleviatory effects in BALB/c mice,which relieved pathological symptoms and lowered production of allergic mediators.Microbial high-throughput sequencing analysis showed that FBP could regulate gut microbiota and metabolism to strengthen immunity,which were closely correlated with the alleviation of allergic reactivity.These findings could contribute to the development and utilization of hypoallergenic BP products.

Microbiomics and metabolomics insights into the microbial regulation on the formation of flavor components in the traditional fermentation process of Chinese Hongqu aged vinegar

This study aimed to investigate microbial succession and metabolic dynamics during the traditional fermentation of Hongqu aged vinegar,and explore the core functional microbes closely related to the formation of flavor components.Microbiome analysis demonstrated that Lactobacillus,Acetobacter,Bacillus,Enterobacter,Lactococcus,Leuconostoc and Weissella were the predominant bacterial genera,while Aspergillus piperis,Aspergillus oryzae,Monascus purpureus,Candida athensensis,C.xylopsoci,Penicillium ochrosalmoneum and Simplicillium aogashimaense were the predominant fungal species.Correlation analysis revealed that Acetobacter was positively correlated with the production of tetramethylpyrazine,acetoin and acetic acid,Lactococcus showed positive correlation with the production of 2-nonanone,2-heptanone,ethyl caprylate,ethyl caprate,1-hexanol,1-octanol and 1-octen-3-ol,C.xylopsoci and C.rugosa were positively associated with the production of diethyl malonate,2,3-butanediyl diacetate,acetoin,benzaldehyde and tetramethylpyrazine.Correspondingly,non-volatile metabolites were also detected through ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry.A variety of amino acids and functional dipeptides were identified during the traditional brewing of Hongqu aged vinegar.Correlation analysis revealed that Lactobacillus was significantly associated with DL-lactate,indolelactic acid,D-(+)-3-phenyllactic acid,pimelic acid,pregabalin and 3-aminobutanoic acid.This study is useful for understanding flavor formation mechanism and developing effective strategies for the suitable strains selection to improve the flavor quality of Hongqu aged vinegar.

Insights into Fermentation Technology for Traditional Chinese Medicine:Progress and Applications

Traditional Chinese medicine(TCM)has an exemplary role in the treatment and prevention of diseases.However,the advancement of TCM has been constrained by several factors,including its intricate structure,low active ingredient concentration,prolonged growth cycle,and the dfficulty in artificial cultivation.In recent years,research on the fermentation technology of TCM has increased.This paper provides an overview of the advantages of TCM fermentation technology,including efficiency enhancement and toxicity reduction,the development of new drugs,and the secondary utilization of dregs.Additionally,it discusses the progress of research on the application of fermented TCM in animal husbandry and disease treatment.The aim is to provide theoretical guidance for TCM fermentation technology research and the development of fermented TCM.

Study on Solid Fermentation and Antioxidant Function of Natto

[Objectives]To study the optimum conditions of solid fermentation of natto with antioxidant function as an index.[Methods]Single factor experiment and orthogonal experiment were designed to study the effects of temperature,time,initial pH and inoculum amount on the antioxidant activity of natto solid fermentation.The optimum conditions of natto solid fermentation were determined and the antioxidant ac-tivity of natto extract was compared.[Results]The optimal fermentation conditions were as follows:temperature 32℃,initial pH 7.0,inocu-lation amount 8%,fermentation time 32 h.The hydroxyl radical scavenging rate of natto solid fermentation crude extract was the highest,which was 82.7%.The optimized nato fermentation extract showed stronger scavenging ability for-OH and O,:,and showed obvious dose-effect relationship.ICso was 3.63 and 4.24 mg/mL,respectively,and the scavenging efficiency was 1.3 and 1.9 times higher than that of the unoptimized fermentation extract,respectively.[Conclusions]Natto is rich in nattokinase and other functional factors,and its antioxidant ac-tivity can be improved by optimizing fermentation technology,so that natto products can be widely used,including cosmetic raw materials,nat-to skin care soap,health food and medicine,etc.,and have a broader development prospect.

Investigation of the Fermentation Mode of Rice Husk for the Stabilization of Earth Plaster

Despite its low resistance to humidity, adobe remains the most widely used material for housing construction, particularly in developing countries. The present study aims to assess different modes of use of fermented RH and to evaluate their influence on the behavior of raw earth for application in plaster. The influences of two types of RH are evaluated: granular rice husk (RHg) and powdered RH (RHp). The clay mainly consists of clay (40%), silt (22%), and sand (38.4%), with a small proportion of gravel (0.24%). Its liquidity limit is 40% and the plasticity index is 26.5%. The mixtures were designed using earth and each of the two rice husks at the volumetric content of 10%, 15% and 20% of the total volume mixed with water 36.5%, 38.5% and 40.3% and fermented for three weeks. Each fermented mixture was added to the soil to form the paste, and 40 × 40 × 160 mm3 test speciments were made for characterization. The results generally show an improvement in the physico-mechanical properties and water resistance of the mortars containing fermented RH, with an optimal content between 10% and 15%. The powdered RH improved the performance of the mortar better than granular RH.

Effect of cooked rice with added fructo-oligosaccharide on faecal microorganisms investigated by in vitro digestion and fermentation

In the present study,the effects of cooked rice(CR)with added fructo-oligosaccharide(FOS)on faecal flora were studied by a simulated in vitro digestion and fermentation method.The total carbohydrate content,p H,and s hort-chain fatty acids(SCFAs)were determined during in vitro digestion and fermentation.The change in the bacterial phase distribution after the fermentation was also analysed.The results showed that t he total carbohydrate content of the CR with added FOS(FCR)significantly decreased during the simulated digestion.Meanwhile,the p H of the FCR decreased and the SCFAs concentration increased significantly compared to those of the CR during the simulated fermentation.In addition,the FCR showed the advantage of promoting beneficial bacteria,such as Bifidobacterium and Lactobacillus,and inhibiting harmful bacteria,such as Bacteroides and Klebsiella compared to the CR.Therefore,the FOS as a prebiotic could be recommended to produce the high-quality healthy rice food.

Artificial simulated saliva,gastric and intestinal digestion and fermentation in vitro by human gut microbiota of intrapolysaccharide from Paecilomyces cicadae TJJ1213

In this study,the in vitro digestion and fermentation of two intra-polysaccharide fractions(IPS1 and IPS2)from Paecilomyces cicadae TJJ1213 were investigated.The constituent monosaccharides of IPS1 and IPS2 were not changed after simulated saliva,gastric and small intestinal digestion.However,they can be hydrolyzed and utilized by gut microbiota,and short-chain fatty acids(SCFAs)level were increased after IPS1 and IPS2 treatments.Furthermore,16 S r RNA sequencing analysis of fermentation samples were performed.Alpha-diversity,beta-diversity and taxonomic composition differences analysis revealed that IPS1 and IPS2 promoted the proliferation of beneficial bacteria and modulated the overall structure of gut microbiota.Taxonomic comparison analysis found that IPS1 increased the relative abundances of beneficial bacteria including Megamonas,Bifidobacterium and Lactobacillus,while IPS2 could increase the abundance of Bacteroides,Parabacteroides and Phascolarctobacterium.In addition,they can also decrease the levels of pathogenic bacteria containing Escherichia-Shigella,Klebsiella and Fusobacterium.These results indicated that IPS from Paecilomyces cicadae TJJ1213 could be used as potential candidates for new functional foods.

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.