Brewer's Grain Research Articles

Phytotoxicity Assessment of Agro-Industrial Waste and Its Biochar: Germination Bioassay in Four Horticultural Species

This study investigated the phytotoxicity of agro-industrial wastes (almond, walnut, pistachio and peanut shells, asparagus spears, and brewer’s spent grain) and their biochar through germination bioassays in several horticultural species: green pea, lettuce, radish, and arugula. Biowaste was pyrolyzed under controlled conditions to produce biochar, and both biowaste and biochar were characterized. Germination bioassay was conducted using seeds exposed to different dilutions of aqueous extract of biowaste and their biochar (0, 50, and 100%). Germination percentage, seed vigor, germination index, and root and aerial lengths were evaluated. The results showed that the phytotoxicity of the biowaste was significantly different to that of its biochar. The biochar obtained demonstrated changing effects on germination and seedling growth. In particular, biochar extracts from spent brewers grains, walnut shells, and pistachio shells showed 5–14% increases in seed vigor and root and aerial length. Furthermore, the response of different species to both agro-industrial waste and biochar revealed species-specific sensitivity. Seeds of lettuce and arugula species were more sensitive to aqueous extracts than radish and green peas. This knowledge not only elucidates the behavior of agro-industrial waste-based biochar in the early stage of plant development but also provides valuable insights regarding phytotoxicity, seed sensitivity, and the variables involved in germination.

Isolation and Structural Characterization of Natural Deep Eutectic Solvent Lignin from Brewer's Spent Grains.

Brewer's spent grains (BSG) offer valuable opportunities for valorization beyond its conventional use as animal feed. Among its components, lignin-a natural polymer with inherent antioxidant properties-holds significant industrial potential. This work investigates the use of microwave-assisted extraction combined with acidic natural deep eutectic solvents (NaDESs) for efficient lignin recovery, evaluating three different NaDES formulations. The results indicate that choline chloride-lactic acid (ChCl-LA), a NaDES with superior thermal stability as confirmed via thermogravimetric analysis (TGA), is an ideal solvent for lignin extraction at 150 °C and 15 min, achieving a balance of high yield and quality. ChCl-LA also demonstrated good solubility and cell disruption capabilities, while microwaves significantly reduced processing time and severity. Under optimal conditions, i.e., 150 °C, 15 min, in the presence of ChCl-LA NaDES, the extracted lignin achieved a purity of up to 79% and demonstrated an IC50 (inhibitory concentration 50%) of approximately 0.022 mg/L, indicating a relatively strong antioxidant activity. Fourier transform infrared (FTIR) and 2D-HSQC NMR (heteronuclear single quantum coherence nuclear magnetic resonance) spectroscopy confirmed the successful isolation and preservation of its structural integrity. This study highlights the potential of BSG as a valuable lignocellulosic resource and underscores the effectiveness of acidic NaDESs combined with microwave extraction for lignin recovery.

The effects of applying cellulase and laccase on fermentation quality and microbial community in mixed silage containing corn stover and wet brewer's grains.

The purpose of this experiment was to explore the effect of adding cellulase and laccase on fermentation quality and microbial community in mixed silage of corn stover and wet brewer's grains. Try to a new approach for the proper preservation and utilization of the agro-industrial by-products (corn stover and wet brewer's grains). The experiment was divided into four groups: CK (control), C (cellulase, 120 U/g fresh matter [FM]), L (laccase, 50 U/g FM), CL (cellulase 120 U/g FW and laccase 50 U/g FM), and the chemical composition, fermentation quality, microbial population and microbial community in mixed silage of corn stover and wet brewer's grains after 30 day's fermentation were determined. Compared to control, the addition of cellulase significantly increased crude protein (CP), water-soluble carbohydrate (WSC), lactic acid bacteria (LAB) counts, while significantly decreased the neutral detergent fiber (NDF), acid detergent fiber (ADF) content (P < 0.05). Adding laccase significantly decreased the acid detergent lignin (ADL) content (P < 0.05). Combined application of cellulase and laccase significantly increased the CP, WSC content and LAB counts, while significantly decreased pH value, NDF, ADF and ADL content (P < 0.05), thereby improving fermentation quality. In addition, the application of cellulase and laccase increased the abundance of Firmicutes and LAB genera, and decreased microbial diversity level of the sample. The combined application of cellulase and laccase further improved fermentation quality and microbial community in mixed silage of corn stover and wet brewer's grains.

Effect of biochars on the concentration of plant-available elements in the soil

To solve environmental problems and reduce economic costs, it is necessary to improve mineral fertilizer application systems by developing and introducing new technologies, including the use of biochars. Due to the lack of effective monitoring of soil changes, it is difficult to adjust fertilizer application rates. This determines the importance of information on the elemental composition of biochar ameliorants used to reduce the mobility of heavy metals in the soil. The use of biochar for remediation of contaminated soils is based on its ability to immobilize heavy metals and organic pollutants. In this work biochars were used, obtained by slow pyrolysis of organic materials of different types: pine sawdust, cattle manure, wheat straw, pine nut shells and brewer's grains. The effect of biochars (biochar application at a dose of 10 t ha−1) on the concentration of 13 elements (C, N, K, P, Mg, Ca, Fe, Mn, Pb, Ni, Cr, Cd, Co) in the soil was analyzed. The data collected after growing of spring wheat (Triticum aestivum L.) on the sod-podzolic soils were analyzed. It has been established that the heavy metals concentrations in the studied soils reclaimed with biochars are significantly lower than the maximum allowable concentrations for soils (MAC). The content of lead (Pb) in soils treated with biochars is 9 or more times lower than the MAC level, and biochar, produced from manure, significantly reduces the concentration of this metal in the soil.

Integrated Metabolomics and Metagenomics Unveiled Biomarkers of Antioxidant Potential in Fermented Brewer's Grains.

About one-third of the global food supply is wasted. Brewers' spent grain (BSG), being produced in enormous amounts by the brewery industry, possesses an eminence nutritional profile, yet its recycling is often neglected for multiple reasons. We employed integrated metagenomics and metabolomics techniques to assess the effects of enzyme treatments and Lactobacillus fermentation on the antioxidant capacity of BSG. The biotreated BSG revealed improved antioxidant capability, as evidenced by significantly increased (p < 0.05) radical scavenging activity and flavonoid and polyphenol content. Untargeted metabolomics revealed that Lactobacillus fermentation led to the prominent synthesis (p < 0.05) of 15 novel antioxidant peptides, as well as significantly higher (p < 0.05) enrichment of isoflavonoid and phenylpropanoid biosynthesis pathways. The correlation analysis demonstrated that Lactiplantibacillus plantarum exhibited strong correlation (p < 0.05) with aucubin and carbohydrate-active enzymes, namely, glycoside hydrolases 25, glycosyl transferases 5, and carbohydrate esterases 9. The fermented BSG has potential applications in the food industry as a culture medium, a functional food component for human consumption, and a bioactive feed ingredient for animals.

Anti-inflammatory Effect of Brewer's Spent Grain In Vitro

Anti-inflammatory Effect of Brewer's Spent Grain In Vitro

ВЛИЯНИЕ ПИВНОЙ ДРОБИНЫ НА ПРОДУКТИВНЫЕ ПОКАЗАТЕЛИ ЦЫПЛЯТ-БРОЙЛЕРОВ КРОССА "РОСС-308"

The research on brewing waste and its use as feed ingredients, in particular for feeding poultry, is a current trend in animal husbandry. Finding new opportunities to reduce the cost of livestock products is an important issue, because The high cost of food is currently becoming a serious problem for consumers. For this reason, studying the possibility of using barley grains in feeding broiler chickens of the Ross-308 cross at JSC PR Mikhailovskoye in the Prigorodny district of North Ossetia-Alania turned out to be very timely. During the experiments, the main production indicators were studied. The results were studied and processed in detail. We determined the effect of brewer’s grain on the health of chickens, on feed intake, their growth and development, and, mainly, on their slaughter performance. According to the approved methodology, 4 groups of day-old chickens were formed. All young animals met the requirements for exterior and constitution. One of them became the control, and the rest became the experimental ones. The first group of broilers was fed the usual feed mixture, which is used on the farm for young animals according to age, compiled according to the current VNITIP standards. The experimental groups were administered barley grains in different quantities - 5 and 7% of the dry matter of the feed. In addition, for these groups, the food was also enriched with the probiotic premix Cellobacterin-T, which replenishes the low amino acid content. The results of the experiments showed that chickens that received 7% barley grains from the dry matter of feed enriched with Cellobacterin-T had better growth and development compared to both the control and experimental groups that received 5% brewers grains, and the one that received feed mixture enriched with premix, without spent grains. Such results give every reason to assert that barley grain is an effective feed ingredient that has a beneficial effect on the body of chickens and stimulates growth and development.

Using brewer's grains correctly

Using brewer's grains correctly

Effects of Alkaline Extraction pH on Amino Acid Compositions, Protein Secondary Structures, Thermal Stability, and Functionalities of Brewer's Spent Grain Proteins.

A high alkaline pH was previously demonstrated to enhance the extraction yield of brewer's spent grains (BSG) proteins. The effects of extraction pH beyond the extraction yield, however, has not been investigated before. The present work examined the effects of extraction pH (pH 8-12) on BSG proteins' (1) amino acid compositions, (2) secondary structures, (3) thermal stability, and (4) functionalities (i.e., water/oil holding capacity, emulsifying, and foaming properties). The ideal extraction temperature (60 °C) and BSG-to-solvent ratio (1:20 w/v) for maximizing the extraction yield were first determined to set the conditions for the pH effect study. The results showed that a higher extraction pH led to more balanced compositions between hydrophilic and hydrophobic amino acids and higher proportions of random coils structures indicating increased protein unfolding. This led to superior emulsifying properties of the extracted proteins with more than twofold improvement between pH 8 and a pH larger than 10. The extraction pH, nevertheless, had minimal impact on the water/oil holding capacity, foaming properties, and thermal denaturation propensity of the proteins. The present work demonstrated that a high alkaline pH at pH 11-12 was indeed ideal for both maximizing the extraction yield (37-46 wt.%) and proteins' functionalities.

FIBRE EFFECTS ON NUTRITION AND REPRODUCTION IN PIGS

Dietary fibre (DF) is an indigestible carbohydrate in the form of a non-starch polysaccharide that is a major problem in the use of inexpensive agro-wastes by pigs such as brewer's dry grains, cassava peel, palm kernel cake, rice bran, sorghum spent grain and wheat offal. The DF in a pig's diet can be classified by its solubility as soluble dietary fibre and insoluble dietary fibre, and chemically − as neutral detergent fibre and acid detergent fibre. High DF levels reduced nutrient utilization, growth rate, in terms of pork production and reproductive performance. Pigs' reproductive cycles at different stages and lactation were both greatly impacted by increased DF intake. High DF decreased the number of stillborn piglets at birth and the farrowing duration by sow, while improving colostrum intake in piglets and colostrum yield in sows. On the other hand, reducing the quantity of DF's hemicellulose in a sow's lactation diet can improve the low-birth-weight piglets' health, post-weaning performance and milk composition. While exogenous enzymes and the extrusion of fibrous feedstuffs enhanced nutrient digestibility and utilisation, other processing methods, including fermentation and extrusion, decreased the DF components and had a beneficial impact on the performance of the pigs. Agro-industrial residues have enormous potential to improve reproductive efficiency and serving as an economically viable DF feeding strategy in the pig industry.

High-Temperature Hydrothermal Extraction of Phenolic Compounds from Brewer's Spent Grain and Malt Dust Biomass Using Natural Deep Eutectic Solvents.

Aligned with the EU Sustainable Development Goals 2030 (EU SDG2030), extensive research is dedicated to enhancing the sustainable use of biomass waste for the extraction of pharmaceutical and nutritional compounds, such as (poly-)phenolic compounds (PC). This study proposes an innovative one-step hydrothermal extraction (HTE) at a high temperature (120 °C), utilizing environmentally friendly acidic natural deep eutectic solvents (NADESs) to replace conventional harmful pre-treatment chemicals and organic solvents. Brewer's spent grain (BSG) and novel malt dust (MD) biomass sources, both obtained from beer production, were characterized and studied for their potential as PC sources. HTE, paired with mild acidic malic acid/choline chloride (MA) NADES, was compared against conventional (heated and stirred maceration) and modern (microwave-assisted extraction; MAE) state-of-the-art extraction methods. The quantification of key PC in BSG and MD using liquid chromatography (HPLC) indicated that the combination of elevated temperatures and acidic NADES could provide significant improvements in PC extraction yields ranging from 251% (MD-MAC-MA: 29.3 µg/g; MD-HTE-MA: 103 µg/g) to 381% (BSG-MAC-MA: 78 µg/g; BSG-HTE-MA: 375 µg/g). The superior extraction capacity of MA NADES over non-acidic NADES (glycerol/choline chloride) and a traditional organic solvent mixture (acetone/H2O) could be attributed to in situ acid-catalysed pre-treatment facilitating the release of bound PC from lignin-hemicellulose structures. Qualitative 13C-NMR and pyro-GC-MS analysis was used to verify lignin-hemicellulose breakdown during extraction and the impact of high-temperature MA NADES extraction on the lignin-hemicellulose structure. This in situ acid NADES-catalysed high-temperature pre-treatment during PC extraction offers a potential green pre-treatment for use in cascade valorisation strategies (e.g., lignin valorisation), enabling more intensive usage of available biomass waste stream resources.

Mixed ensiling of drought‐impaired grass with agro‐industrial by‐products and silage additives improves the nutritive value and shapes the microbial community of silages

AbstractDroughts lead to reduced biomass production and unfavourable nutrient composition in grassland. As an alleviation, yet unexploited strategy, mixed ensiling of grass with agro‐industrial by‐products may improve the ensilability and nutritive value of drought‐impaired grassland. This study investigated first whether mixed ensiling of drought‐impaired grass with either sugar beet pulp (SBP), wheat gluten feed (WGF) or brewers' grains (BG) has a beneficial impact on chemical composition, fermentation characteristics, in vitro gas production (GP) and physically effective neutral detergent fibre (peNDF) of silages. Secondly, it was tested whether the application of anaerobic fungi culture supernatant (AF), mixed ruminal fluid (RF) or lactic acid bacteria (LAB) provides further advantages. Additionally, the microbial community composition was evaluated in selected silages. All silages showed satisfying conservation characteristics with high lactic acid levels and low dry matter losses, and peNDF values typically found for conserved forages. Mixed ensiling with BG substantially increased the crude protein concentration, whereas SBP increased the total degradability and WGF enhanced both. The further addition of fresh AF resulted in the overall highest lactic acid levels, especially in SBP‐based silages, but without changes in in vitro GP. The in vitro GP was higher with RF, particularly in mixed silages, suggesting an improved degradability. The LAB‐treated silages showed lower pH compared to controls, but had no impact on in vitro GP kinetics. Concluding, mixed ensiling holds potential to produce high‐quality silages from drought‐impaired grassland. The further addition of silage additives can be useful for certain substrates, but appeared not mandatory.

Rice Husk, Brewer's Spent Grain, and Vine Shoot Trimmings as Raw Materials for Sustainable Enzyme Production.

Solid by-products with lignocellulosic structures are considered appropriate substrates for solid-state fermentation (SSF) to produce enzymes with diverse industrial applications. In this work, brewer's spent grain (BSG), rice husk (RH), and vine shoot trimmings (VSTs) were employed as substrates in SSF with Aspergillus niger CECT 2088 to produce cellulases, xylanases, and amylases. The addition of 2% (NH4)2SO4 and 1% K2HPO4 to by-products had a positive effect on enzyme production. Substrate particle size influenced enzyme activity and the overall highest activities were achieved at the largest particle size (10 mm) of BSG and RH and a size of 4 mm for VSTs. Optimal substrate composition was predicted using a simplex centroid mixture design. The highest activities were obtained using 100% BSG for β-glucosidase (363 U/g) and endo-1,4-β-glucanase (189 U/g), 87% BSG and 13% RH for xylanase (627 U/g), and 72% BSG and 28% RH for amylase (263 U/g). Besides the optimal values found, mixtures of BSG with RH or VSTs proved to be alternative substrates to BSG alone. These findings demonstrate that SSF bioprocessing of BSG individually or in mixtures with RH and VSTs is an efficient and sustainable strategy to produce enzymes of significant industrial interest within the circular economy guidelines.

ФИЗИКО-ХИМИЧЕСКИЕ СВОЙСТВА ФЕРМЕНТИРОВАННЫХ КОМБИКОРМОВ В ЗАВИСИМОСТИ ОТ СПОСОБА ПРИГОТОВЛЕНИЯ

The article presents the results of a comparative study of the physicochemical parameters of fermented compound feed consisting of 40% brewer's grains, 20% wheat bran, 20% sunflower cake, 20% mushroom substrate. Prepared in two ways: the first way – the compound feed composed of native ingredients was fermented, the second – the compound feed composed of pre-fermented ingredients. In the compound feed composed of fermented ingredients, in comparison with the compound feed fermented in full, the mass fraction of moisture, crude fat, soluble carbohydrates, starch and crude fiber was higher; the mass fraction of crude protein and exchange energy was lower.

Sequential Extraction and Attenuated Total Reflection-Fourier Transform Infrared Spectroscopy Monitoring in the Biorefining of Brewer's Spent Grain.

The brewing industry plays a significant role in producing a substantial annual volume of by-products, which contributes to the global accumulation of food waste. The primary by-product generated is brewer's spent grain (BSG), a lignocellulosic biomass rich in proteins, fiber, and moisture content. Leveraging biorefining and valorization techniques for BSG represents a promising strategy to enhance sustainability, resilience, and circularity within the brewing chain. To date, most studies have focused on extracting proteins from BSG. Yet, it is crucial to note that the fiber part of BSG also holds considerable potential for biorefining processes. This study introduces a novel sequential extraction method designed to integrally recover the major components of BSG. Notably, it introduces a reactive extraction approach that enables the simultaneous extraction and tuneable functionalization of the hemicellulose component. Additionally, the study assesses the utility of the attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy as a user-friendly tool to monitor and evaluate the effectiveness of the fractionation process. This spectroscopic technique can provide valuable insights into the changes and composition of BSG throughout the extraction process.

Versatile Applications of Brewer's Spent Grain: Solid-State Fermentation and Nutritional Added Value.

Brewer's spent grain (BSG) is a major by-product in the beer-brewing process which contributes to 85% of the entire generated by-product in the brewing process. BSG is rich in proteins, and most of the malt proteins (74-78%) remain insoluble in BSG after the mashing process. Solid-state fermentation (SSF) is a promising bioprocess that enables microorganisms to survive in environments with minimal water and has shown to enhance the nutritional composition of BSG. In this review, the potential application of protein, amino acids (proline, threonine, and serine), phenolic contents, and soluble sugars (glucose, fructose, xylose, arabinose, and cellobiose) extracted from BSG by various microorganisms using SSF is explored. Incorporation of BSG into animal feed, human diets, and as a substrate for microorganisms are the prospects that could be implemented in the industrial scale. This review also discussed various advances to improve the fermentation yield such as symbiotic fermentation, the addition of nitrogen supplements, and an optimal mixture of the agro-industrial waste substrate. Future perspectives on SSF are also addressed to provide important ideas for immediate and future studies. However, challenges include optimizing SSF conditions and design of bioreactors, and operational costs must be addressed in the future to overcome current obstacles. Overall, this mini review highlights the potential benefits of BSG utilization and SSF in a sustainable way.

358 The Effects of Varying Levels of Cotton Gin Byproduct and Wet brewers’ Grains on Growth Performance of Growing and Finishing Steers

Abstract The abundance of craft breweries provides widespread availability of wet brewers grains (WBG) to be used by beef cattle producers as a potential feedstuff. Cotton gin byproduct (CGB) is a feedstuff typically reserved for mature beef cows due to its low and variable feed value. Both WBG and CGB have high cost of transportation but are sometimes used by producers near supply centers due to their low price. The objective of this study was to analyze animal performance of growing Angus steers fed total mixed rations with varying amounts of CGB and WBG. Sixty-four Angus steers (365 ± 26 kg) were allotted and blocked by initial body weight (BW) in a randomized complete block design to 8 dietary treatments and fed in 2 phases, growing phase (GP) and finishing phase (FP) (84-d and 69-d, respectively). Dietary treatments were arranged in a 2 x 4 factorial with two levels of CGB (0 or 25% of DM) and 4 levels of WBG (0, 10, 20 or 30% of DM). The control and base of all treatments were corn, corn silage and soybean meal. Body weights were recorded on two consecutive days at the beginning and end of GP and FP. Feed samples were collected, and refusals were weighed weekly. Data were analyzed using the MIXED procedure of SAS 9.4 with main effects of CGB, WBG, and the interaction. The linear, quadratic, and cubic effects of increasing levels of WBG and CGB were tested with polynomial contrasts. Average daily gain (ADG) did not differ among dietary treatments (0, 25% CGB and 0, 10, 20, 30% WBG) for GP, (1.67, 1.61, and 1.67, 1.57, 1.74, and 1.60 kg/d, respectively) (P &amp;gt; 0.3031), FP (1.53, 1.61 and 1.56, 1.53, 1.61, and 1.58 kg/d) or the overall trial (1.61, 1.62 and 1.62, 1.55, 1.68, and 1.60 kg/d). The addition of CGB increased GP dry matter intake (DMI; 11.06 vs 11.99 kg/d; P = 0.0011), while the addition of WBG linearly decreased GP DMI (11.81, 11.69. 11.81, and 10.81 kg/d; P = 0.0292). There was an interaction between CGB and WBG (P = 0.0206), with increasing levels of WBG at 0% CGB exhibiting a cubic effect (P = 0.0127), but no effect at 25% CGB. Inclusion of CGB decreased feed efficiency (FE) in the GP (6.77 vs. 7.69; P = 0.0008) but had no carry-over effect in the FP. There was no effect of WBG on FE in GP or FP. In conclusion, CGB increased DMI, reduced FE, and maintained ADG. Increasing quantities of WBG did not reduce ADG or FE, but decreased DMI with increasing feeding level. Therefore, WBG and CGB can be considered viable options for cattle producers seeking to lower input costs.

Influence of Hydrothermal Treatment of Brewer's Spent Grain on the Concentration and Molecular Weight Distribution of 1,3-1,4-β-D-Glucan and Arabinoxylan.

Brewer's spent grain (BSG) is the most abundant residual in the brewing process. Non-starch polysaccharides such as 1,3-1,4-β-D-glucan (β-glucan) and arabinoxylan (AX) with proven beneficial effects on human health remain in this by-product in high amounts. Incorporating the valuable dietary fiber into the food industry could contribute to a healthy diet. However, a major challenge is extracting these dietary fibers (i.e., β-glucan and AX) from the solid residue. In this study, hydrothermal treatment (HT) was applied to dissolve the remaining water-insoluble carbohydrates from BSG with the aim to extract high amounts of β-glucan and AX. Particular focus was placed on the molecular weight (MW) range above 50 kDa and 20 kDa, respectively, as these are considered to have health-promoting effects. Different treatment temperatures, reaction times, and internal reactor pressures were tested to determine the best process settings to achieve high yields of β-glucan and AX and to examine the influence on their molecular weight distribution (MWD). Overall, 85.1% β-glucan and 77.3% AX were extracted corresponding to 6.3 g per kg BSG at 160 °C and 178.3 g kg-1 at 170 °C, respectively. However, less than 20% of both fiber substances were in the desirable MW range above 50 kDa and 20 kDa, respectively. When lower temperatures of 140 and 150 °C were applied, yields of only 3.0 g kg-1 β-glucan and 128.8 g kg-1 AX were obtained, whereby the proportion of desirable fiber fractions increased up to 45%. Further investigations focused on the heat-induced degradation of monosaccharides and the formation of undesirable by-products (i.e., HMF and furfural) that might pose a health risk.

INNOVATIVE AND SUSTAINABLE MULTI-SERVICE MODELS TO BOOST LIVESTOCK FARMING SYSTEM

INNOVATIVE AND SUSTAINABLE MULTI-SERVICE MODELS TO BOOST LIVESTOCK FARMING SYSTEM

Fermentation characteristics and microbial community composition of wet brewer's grains and corn stover mixed silage prepared with cellulase and lactic acid bacteria supplementation.

The objective of this study was to investigate how cellulase or/and lactic acid bacteria (LAB) affected the fermentation characteristic and microbial community in wet brewer's grains (WBG) and corn stover (CS) mixed silage. The WBG was mixed thoroughly with the CS at 7:3 (w/w). Four treatment groups were studied: i) CON, no additives; ii) CEL, added cellulase (120 U/g fresh matter [FM]), iii) LAB, added LAB (2×106 cfu/g FM), and iv) CLA, added cellulase (120 U/g FM) and LAB (2×106 cfu/g FM). All additive-treated groups showed higher fermentation quality over the 30 d ensiling period. As these groups exhibited higher (p<0.05) LAB counts and lactic acid (LA) content, along with lower pH value and ammonia-nitrogen (NH3-N) content than the control. Specifically, cellulase-treated groups (CEL and CLA) showed lower (p<0.05) neutral detergent fiber and acid detergent fiber contents than other groups. All additives increased the abundance of beneficial bacteria (Firmicutes, Lactiplantibacillus, and Limosilactobacillus) while they decreased abundance of Proteobacteria and microbial diversity as well. The combined application of cellulase and LAB could effectively improve the fermentation quality and microbial community of the WBG and CS mixed silage.