Effects of non-pelleted or pelleted low-native grass and pelleted high-native grass diets on meat quality by regulating the rumen microbiota in lambs.

Diet modulates the rumen microbiota, which in turn can impact the animal performance. The rumen microbiota is increasingly recognized for its crucial role in regulating the growth and meat quality of the host. Nevertheless, the mechanism by which the rumen microbiome influences the fatty acid and amino acid profiles of lambs in the grass feeding system remains unclear. This study aimed to evaluate the effects of different native grass-based diets on animal performance, meat quality, fatty acid compositions, amino acid profiles, and rumen microbiota of lamb. Seventy-two Ujumqin lambs were randomly assigned into three treatments according to the initial body weight (27.39 ± 0.51 kg) and age (6 months ± 6 days). The lambs received three diets: (i) non-pelleted native grass hay with 40% concentrate diet; the native grass and concentrate were fed individually; (ii) pelleted native grass hay with 40% concentrate diet (PHLC); (iii) pelleted native grass hay with 60% concentrate diet (PHHC). The results showed that among the three groups, the PHHC and PHLC diets had markedly (P < 0.05) higher average daily gain and pH45 min, respectively. All amino acid levels were significantly (P < 0.05) decreased in the PHHC diet than in the PHLC diet. The principal coordinate analysis of the ruminal microbiota indicated the markedly distinct separation (P = 0.001) among the three groups. In addition, the correlation analysis showed that the Rikenellaceae_RC9_gut_group, Prevotellaceae_UCG-003, Succinivibrio, and Succiniclasticum were significantly (P < 0.05) associated with most of the fatty acid and amino acid profiles. The correlation analysis of the association of microbiome with the meat quality provides us with a comprehensive understanding of the composition and function of the rumen microbial community, and these findings will contribute to the direction of future research in lamb. IMPORTANCE: Diet modulates the gut microbiome, which in turn impact the meat quality, yet few studies investigate the correlation between the rumen microbiome and the fatty acid profile of meat. Here, the current study develops an experiment to investigate the correlation of the rumen microbiome and fatty acid profile of meat: rumen microbiome responses to feed type and meat quality. The results indicated a unique microbiota in the rumen of lamb in response to diets and meat quality. Associations between utilization and production were widely identified among the affected microbiome and meat quality, and these findings will contribute to the direction of future research in lamb.

Degradation of algae promotes the release of arsenic from sediments under high-sulfate conditions.

Sulfate concentrations in eutrophic waters continue to increase; however, the transformations of arsenic (As) in sediments under these conditions are unclear. In this study, we constructed a series of microcosms to investigate the effect of algal degradation on As transformations in sediments with high sulfate concentrations. The results showed that both the elevated sulfate levels and algal degradation enhanced the release of As from sediments to the overlying water, and degradation of algal in the presence of elevated sulfate levels could further contribute to As release. Sulfate competed with arsenate for adsorption in the sediments, leading to As desorption, while algal degradation created a strongly anaerobic environment, leading to the loss of the redox layer in the surface sediments. With high sulfate, algal degradation enhanced sulfate reduction, and sulfur caused the formation of thioarsenates, which may cause re-dissolution of the arsenides, enhancing As mobility by changing the As speciation. The results of sedimentary As speciation analysis indicated that elevated sulfur levels and algal degradation led to a shift of As from Fe2O3/oxyhydroxide-bound state to specifically adsorbed state at the sediment water interface. This study indicated that algal degradation increases the risk of As pollution in sulfate-enriched eutrophic waters.

Integrated rumen microbiome and serum metabolome analysis responses to feed type that contribution to meat quality in lambs.

BACKGROUND: Lifestyle factors, such as diet, are known to be a driver on the meat quality, rumen microbiome and serum metabolites. Rumen microbiome metabolites may be important for host health, the correlation between rumen microbiome and production of rumen metabolites are reported, while the impact of rumen microbiome on the serum metabolome and fatty acid of meat are still unclear. This study was designed to explore the rumen microbiome, serum metabolome and fatty acid of meat in response to the grass diet and concentrate diet to lambs, and the relationship of which also investigated. METHODS: In the present study, 12 lambs were randomly divided into two groups: a grass diet (G) and a concentrate diet (C). Here, multiple physicochemical analyses combined with 16S rRNA gene sequences and metabolome analysis was performed to reveal the changes that in response to feed types. RESULTS: The concentrate diet could improve the growth performance of lambs compared to that fed with the grass diet. The microbiome composition was highly individual, compared to the concentrate group, the abundance of Rikenellaceae_RC9_gut_group, F082_unclassified, Muribaculaceae_unclassified, Ruminococcaceae_NK4A214_group, Bacteroidetes_unclassified, and Bacteroidales_UCG-001_unclassified were significantly (P < 0.05) lower in the grass group, while, the abundance of Succinivibrio, Succinivibrionaceae_UCG-002, Fibrobacter and Christensenellaceae_R-7_group were significantly (P < 0.05) higher in the grass group. Serum metabolomics analysis combined with enrichment analysis revealed that serum metabolites were influenced by feed type as well as the metabolic pathway, and significantly affected serum metabolites involved in amino acids, peptides, and analogues, bile acids, alcohols and derivatives, linoleic acids derivatives, fatty acids and conjugates. Most of the amino acids, peptides, and analogues metabolites were positively associated with the fatty acid contents. Among the bile acids, alcohols and derivatives metabolites, glycocholic was positively associated with all fatty acid contents, except C18:0, while 25-Hydroxycholesterol and lithocholic acid metabolites were negatively associated with most of the fatty acid contents. CONCLUSION: Correlation analysis of the association of microbiome with metabolite features, metabolite features with fatty acid provides us with comprehensive understanding of the composition and function of microbial communities. Associations between utilization or production were widely identified among affected microbiome, metabolites and fatty acid, and these findings will contribute to the direction of future research in lamb.

Metabolic profiling provides insights into the accumulation patterns of flavonoids and phenolic acids in different parts of Lactuca indica L.

Lactuca indica L. contains high amounts of flavonoids and phenolic acids. However, there is limited information on the composition of these compounds in different parts of the plant. The present study analyzed the secondary metabolite profiles of the stem, leaf, flower, and seed of Lactuca indicaL.cv. Mengzao (LIM) using a widely targeted metabolomic approach. A total of 576 secondary metabolites were identified, including 218, 267, 232, 286, 302, and 308 differentially accumulated metabolites (DAMs) in the stem_vs_leaf, stem_vs_flower, stem_vs_seed, leaf_vs_flower, leaf_vs_seed, and flower_vs_seed comparisons. In particular, considerable differences were detected in the flavonoids and phenolic acids, five flavonoids, five phenolic acids, one triterpenoid and one alkaloid being differentially accumulated in the four parts. Compared to the stem and flower, the leaf and seed had higher total flavonoid content and total phenolic content. Thses findings provide comprehensive insights into utilizing different parts of LIM in developing functional food products.

Chemical profile and quantitative comparison of constituents in different medicinal parts of Lactuca indica during varied harvest periods using UPLC-MS/MS method.

Lactuca indica L. cv. Mengzao (LIM), acknowledged as a pivotal "One Root of Medicine and Food", boasts dual applications in both culinary and medicinal domains. This research delves into the influence of various harvest periods (vegetative, budding, blossom, and fruiting) on distinct medicinal parts (roots, stems, leaves, flowers, and seeds) of LIM, employing plant metabolomics to assess its chemical constituents. A total of 66 chemical constituents were identified in LIM, with 11 chemical components emerging as potential markers for distinguish medicinal parts. Notably, nutritional organs exhibited elevated levels of cichoric acid, rutin and chlorogenic acid. Specifically, leaves during the budding stage displayed the highest chicoric acid content at 11.70 mg·g-1. Conversely, reproductive organs showed heightened concentrations of cichoric acid, rutin and chlorogenic acid, with seeds exhibiting the peak cichoric acid content at 4.53 mg g-1. This study enriches our understanding of LIM by offering novel insights into quality assessment and the comprehensive utilization of its diverse parts.

Microbiomics and volatile metabolomics-based investigation of changes in quality and flavor of oat (Avena sativa L.) silage at different stages.

Objective: This study aimed to analyze the fermentation quality, microbial community, and volatile metabolites of oat silage harvested at two different stages, while examining the correlation between microorganisms and volatile metabolites. Methods: Oats were harvested at two growth stages (pre-heading [PRH] and post-heading [POH] stages), followed by 90 days of natural fermentation, with 6 replicates per treatment. Pre- and post-silage samples were randomly selected for nutrient composition, fermentation parameters, microbial population, and high-throughput sequencing analysis. Volatile metabolomics analysis was also performed on samples after 90 days of fermentation to detect differences in flavor quality after silage. Results: The effect of growth stage on the nutrient content of oats was significant, with pre-heading oats having higher crude protein and post-heading oats having higher water soluble carbohydrates content (p < 0.05). Following a 90-day fermentation period, the pH and ammonia nitrogen/total nitrogen levels in the PRH-90 (silage from pre-heading oats after 90 days of fermentation) group demonstrated a significant decrease (p < 0.05), whereas the lactic acid content was notably higher compared to the POH-90 (silage from post-heading oats after 90 days of fermentation) group (p <0.05). Lactiplantibacillus dominated in the PRH-90 group and Enterococcus dominated in the POH-90 group, with abundances of (> 86%) and (> 87%), respectively. The differential volatile metabolites of the two treatment groups were dominated by esters and terpenoids, and the differences in flavor were mainly concentrated in sweet, green, and fruity odors. The results of Kyoto encyclopedia of genes and genomes pathway enrichment analysis demonstrated three major metabolic pathways: phenylpropanoid biosynthesis, phenylalanine metabolism, and biosynthesis of secondary metabolites. Specific microorganisms were significantly correlated with flavor indicators and flavor metabolites. Lactiplantibacillus was significantly positively correlated with flavor substances indicating sweet and fruity flavors, contributing to good flavor, while Enterococcus was significantly and positively correlated with flavor substances indicating bad flavors. Conclusion: In summary, growth stage had significant effects on nutritional components, fermentation parameters and flavor quality of oats, with the fermentation process dominated by Lactiplantibacillus leading to good flavor, while the fermentation process dominated by Enterococcus led to the development of poor flavor.

Optimization of ultrasonic-assisted extraction of flavonoids from Lactuca indica L. cv. Mengzao and their antioxidant properties.

In this study, the ultrasound-assisted extraction (UAE) conditions of flavonoids from Lactuca indica L.cv. Mengzao (LIM) leaves was optimized, and the flavonoids content and their antioxidant potential in different parts were analyzed. The optimal extraction parameters to obtain the highest total flavonoids content (TFC) were a a ratio of liquid to solid of 24.76 mL/g, ultrasonic power of 411.43 W, ethanol concentration of 58.86% and an extraction time of 30 min, the average TFC of LIM leaves could reach 48.01 mg/g. For the yield of flavonoids, the UAE method had the best extraction capacity compared with solvent extraction and microwave-assisted extraction (MAE). In general, the TFC in different parts of LIM followed the order flower > leaf > stem > root, the flowering period is the most suitable harvesting period. From ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS) quantification, the flower samples showed significantly higher six flavonoids and had the highest radical scavenging capacities compared to other samples. A high positive correlation was observed between the antioxidant activity and TFC, luteolin-7-O-glucoside and rutin were significantly (p < 0.05) correlated with all antioxidant evaluations. This study provides valuable information for the development and utilization of flavonoids in Lactuca indica as ingredients in food, feed and nutritional health products.

Effects of different forage proportions in fermented total mixed ration on muscle fatty acid profile and rumen microbiota in lambs.

Objective: The objectives of this study were to evaluate the effects of different forage proportions in the fermented total mixed ration (FTMR) on growth performance, muscle fatty acid profile, and rumen microbiota of lambs. Methods: Thirty 6-month-old small tail Han sheep × Ujumqin lambs with initial body weight (BW) of 27.8 ± 0.90 kg were selected for the test and divided into two groups of 15 sheep in each treatment (three pens per treatment and five lambs per pen) according to the principle of homogeneity. Two isoenergetic and isonitrogenous diets were formulated according to the NRC. The diet treatments were designed as (1) OH treatment containing 25% alfalfa hay and 35% oat hay, and (2) AH treatment containing 35% alfalfa hay with 25% oat hay. The forage-to-concentrate ratio for both diets was 65: 35 (DM basis). Three replicates were randomly selected from each treatment to determine growth performance, fatty acid profile and rumen bacterial communities in lambs. Results: Results revealed no statistically significant (p > 0.05) differences in dry matter intake and average daily gain between the two diet groups. Cholesterol and intramuscular fat were significantly (p > 0.05) higher in the AH group, while no statistically significant difference (p > 0.05) was found in pH24 value. The muscle fatty acid compositions of lambs were obviously (p < 0.05) influenced by the diet treatments. Compared with the OH group, the C16:1, C17:0, and C20:3n6 contents were higher (p < 0.05) in the AH group, whereas the content of C18:1n9c, C20:1, C18:3n3, and C22:6n3 was obviously (p < 0.05) increased in the OH group. The monounsaturated fatty acid (MUFA) contents were significantly higher in the OH group, whereas no significant differences (p > 0.05) were detected in saturated fatty acid (SFA) and polyunsaturated fatty acid (PUFA) contents among the two diet treatments. Bacterial composition was generally separated into two clusters based on principal coordinate analysis, and the OH group had a higher Shannon index. The relative abundance at the genes level of the Rikenellaceae_RC9_gut_group was obviously (p < 0.05) increased in the AH group and the relative abundances of Prevotella_1, Fibrobacter, and Bacteroidales_UCG_001_unclassified were obviously (p < 0.05) enriched in the OH group. Integrated correlation analysis also underscored a possible link between the muscle fatty acid compositions and significantly altered rumen microbiota. Conclusion: Overall, oat-based roughage in FTMR could promote a beneficial lipid pattern in the Longissimus lumborum muscles of lambs. These findings provide a potential insight into diet effects on fatty acid profile and the rumen microbiome of lambs, which may help make decisions regarding feeding.

Effect of isolated lactic acid bacteria on the quality and bacterial diversity of native grass silage.

Objective: The objective of this study was to isolate lactic acid bacteria (LAB) from native grasses and naturally fermented silages, determine their identity, and assess their effects on silage quality and bacterial communities of the native grasses of three steppe types fermented for 60 days. Methods: Among the 58 isolated LAB strains, Limosilactobacillus fermentum (BL1) and Latilactobacillus graminis (BL5) were identified using 16S rRNA sequences. Both strains showed normal growth at 15- 45°C temperature, 3-6.5% NaCl concentration, and pH 4-9. Two isolated LAB strains (labeled L1 and L5) and two commercial additives (Lactiplantibacillus plantarum and Lentilactobacillus buchneri; designated as LP and LB, respectively) were added individually to native grasses of three steppe types (meadow steppe, MS; typical steppe, TS; desert steppe, DS), and measured after 60 d of fermentation. The fresh material (FM) of different steppe types was treated with LAB (1 × 105 colony forming units/g fresh weight) or distilled water (control treatment [CK]). Results: Compared with CK, the LAB treatment showed favorable effects on all three steppe types, i.e., reduced pH and increased water-soluble carbohydrate content, by modulating the microbiota. The lowest pH was found in the L5 treatment of three steppe types, at the same time, the markedly (p < 0.05) elevated acetic acid (AA) concentration was detected in the L1 and LB treatment. The composition of bacterial community in native grass silage shifted from Pantoea agglomerans and Rosenbergiella nectarea to Lentilactobacillus buchneri at the species level. The abundance of Lentilactobacillus buchneri and Lactiplantibacillus plantarum increased significantly in L1, L5, LP, and LB treatments, respectively, compared with CK (p < 0.05). Conclusion: In summary, the addition of LAB led to the shifted of microbiota and modified the quality of silage, and L. fermentum and L. graminis improved the performance of native grass silage.

Alternations of ensiling performance and bacterial community in response to different native grassland in Mongolian Plateau.

AIMS: The aim of this study was to investigate the dynamics of bacterial communities and natural fermentation quality in three steppe types [meadow steppe (MS); typical steppe (TS); and desert steppe (DS)] on the Mongolian Plateau. METHODS AND RESULTS: PacBio single molecule with real-time sequencing technology was applied to provide insights into the dynamics of the physicochemical characteristics and the complex microbiome of native grass after 1, 7, 15, and 30 days of fermentation process. The dry matter, crude protein, and water soluble carbohydrate (WSC) contents of the three groups slowly decreased after 1 day of fermentation process, and the lowest WSC concentration after 30 days of ensiling was detected in the DS group compared to that in the MS and TS groups. There was no significant effect of steppe types on lactic acid and butyric acid content (P > 0.05). The pH was higher in the early stages of fermentation. After 30 days of fermentation, the pH of MS and DS dropped to ∼5.60, while TS was as high as 5.94. At different ensiling days, the pH of TS was significantly higher than that of MS (P < 0.05). The ammonia nitrogen content of MS was significantly higher than TS and DS (P < 0.05). During the whole fermentation process, Leuconostoc mesenteroides and Pseudocitrobacter faecalis were the main species of DS, while Enterobacter roggenkampii and Faecalibacterium prausnitzii dominated the fermentation process in MS and TS, respectively. CONCLUSIONS: The fermentation quality of native grass silage of different steppe types was less satisfactory, with the silage quality ranging from DS, MS, and TS in descending order. The epiphytic bacteria dominating the fermentation process differed between steppe types of silage. Leuconostoc mesenteroides as the main strain of DS had a modulating effect on pH and LA content, while the main strains of MS and TS (Enterobacter roggenkampii and Faecalibacterium prausnitzii) dominated the silage without significant effect on improving fermentation characteristics and nutritional quality.

Optimization of Extraction Process and Dynamic Changes in Triterpenoids of Lactuca indica from Different Medicinal Parts and Growth Periods.

In this study, the triterpenoids in the leaves of Lactuca indica L.cv. Mengzao (LIM) were extracted via microwave-assisted ethanol extraction, and the optimum extraction conditions for triterpenoids were determined through single-factor experiments and the Box-Behnken method. The effects of three factors (solid-liquid ratio, microwave power and extraction time) on the total triterpenoids content (TTC) were evaluated. The TTC of different parts (roots, stems, leaves and flowers) of LIM in different growth stages was studied, and the scavenging effects of the highest TTC parts on DPPH, ABTS and hydroxyl free radicals were investigated. The results showed that the optimum extraction conditions for microwave-assisted extraction of total triterpenoids from LIM leaves were as follows: solid-liquid ratio of 1:20 g/mL; microwave power of 400 W; and extraction time of 60 min. Under these conditions, the TTC was 29.17 mg/g. Compared with the fresh raw materials, the TTC of the materials increased after freeze drying. The leaves of LIM had the highest TTC, and the flowering stage was the best time. The triterpenoids from the leaves had a strong ability to eliminate DPPH and ABTS free radicals, and the elimination effect of dried leaves was better than that of fresh leaves, while the elimination effect of hydroxyl free radicals was not obvious. The tested method was used to extract total triterpenoids from LIM using a simple process at low cost, which provides a reference for developing intensive processing methods for L. indica.

Effects of Cellulase and Lactic Acid Bacteria on Ensiling Performance and Bacterial Community of Caragana korshinskii Silage.

The aim of this study was to evaluate the effects of cellulase (CE) and lactic acid bacteria (LAB) on Caragana korshinskii silage by analyzing the fermentation parameters, chemical composition, and bacterial community. The Caragana korshinskii was harvested at the fruiting period and treated with cellulase and LAB alone as a control treatment with no additive (CK). The ensiling performance and bacterial community were determined after 3, 7, 15, 30, and 60 days of fermentation process. Compared with the CK group, the pH, dry matter loss, and ammonia nitrogen content were significantly (p < 0.05) decreased in the LAB and CE treatments. Compared with the CK and LAB group, the contents of acid detergent fiber, neutral detergent fiber, and acid detergent lignin in the CE group decreased significantly (p < 0.05), and the water-soluble carbohydrates, acetic acid, and lactic acid concentrations increased significantly (p < 0.05). At the genus level of microorganisms, the addition of cellulase and LAB significantly reduced the microbial diversity. Compared with the CK group (78.05%), the relative abundance of Lactiplantibacillus in the CE group (90.19%) and LAB group (88.40%) significantly (p < 0.05) increased. The relative abundance of Pediococcus in the CE group (3.66%) and LAB group (2.14%) was significantly (p < 0.05) lower than that in the CK group (14.73%). Predicted functional profiling of 16S rRNA genes revealed that the addition of cellulase and LAB increased the pyruvate metabolic pathway during Caragana korshinskii silage, thereby increasing the accumulation of lactic acid concentration. The addition of cellulase expressed a better advantage in the biosynthetic capacity of lysine. In summary, the addition of cellulase and LAB could adjust the bacterial community to improve the silage quality of Caragana korshinskii, and the addition of cellulase exhibited better results than the LAB additives.

Influence of Cellulase or Lactiplantibacillus plantarum on the Ensiling Performance and Bacterial Community in Mixed Silage of Alfalfa and Leymus chinensis.

The objective of this study was to evaluate the effects of Lactiplantibacillus plantarum or cellulase on the fermentation characteristics and bacterial community of mixed alfalfa (Medicago sativa L., AF) and Leymus chinensis (LC) silage. The harvested alfalfa and Leymus chinensis were cut into 1-2 cm lengths by a crop chopper and they were thoroughly mixed at a ratio of 3/2 (wet weight). The mixtures were treated with no addition (CON), Lactiplantibacillus plantarum (LP, 1 × 106 cfu/g fresh material), cellulase (CE, 7.5 × 102 U/kg fresh material) and their combination (LPCE). The forages were packed into triplicate vacuum-sealed, polyethylene bags per treatment and ensiled for 1, 3, 5, 7 and 30 d at room temperature (17-25 °C). Compared to the CON groups, all the additives increased the lactic acid content and decreased the pH and ammonia nitrogen content over the ensiling period. In comparison to the other groups, higher water-soluble carbohydrate contents were discovered in the CE-inoculated silages. Compared to the CON groups, the treatment with LPCE retained the crude protein content and reduced the acid detergent fiber content. The principal coordinate analysis based on the unweighted UniFrac distance showed that individuals in the AF, LC, CON and LPCE treatment could be significantly separated from each other. At the genus level, the bacterial community in the mixed silage involves a shift from Cyanobacteria_unclassified to Lactobacillus. Lactobacillus dominated in all the treatments until the end of the silage, but when added with Lactiplantibacillus plantarum, it was more effective in inhibiting undesirable microorganisms, such as Enterobacter, while reducing microbial diversity. By changing the bacterial community structure after applying Lactiplantibacillus plantarum and cellulase, the mixed silages quality could be further improved. During ensiling, the metabolism of the nucleotide and carbohydrate were enhanced whereas the metabolism of the amino acid, energy, cofactors and vitamins were hindered. In conclusion, the relative abundance of Lactobacillus in the mixed silage increased with the addition of Lactiplantibacillus plantarum and cellulase, which also improved the fermentation quality.

Effects of Different Types of LAB on Dynamic Fermentation Quality and Microbial Community of Native Grass Silage during Anaerobic Fermentation and Aerobic Exposure.

Silage of native grasses can alleviate seasonal forage supply imbalance in pastures and provide additional sources to meet forage demand. The study aimed to investigate the effects of Lactobacillus plantarum (LP), Lactobacillus buchneri (LB), and Lactobacillus plantarum in combination with Lactobacillus buchneri (PB) on the nutritional quality, fermentation quality, and microbial community of native grass silage at 2, 7, 15, and 60 days after ensiling and at 4 and 8 days after aerobic exposure. The results showed that dry matter content, crude protein content, the number of lactic acid bacteria, and lactic acid and acetic acid content increased and pH and ammonia nitrogen content decreased after lactic acid bacteria (LAB) inoculation compared with the control group (CK). LP had the lowest pH and highest lactic acid content but did not have greater aerobic stability. LB maintained a lower pH level and acetic acid remained at a higher level after aerobic exposure; aerobic bacteria, coliform bacteria, yeast, and molds all decreased in number, which effectively improved aerobic stability. The effect of the compound addition of LAB was in between the two other treatments, having higher crude protein content, lactic acid and acetic acid content, lower pH, and ammonia nitrogen content. At the phylum level, the dominant phylum changed from Proteobacteria to Firmicutes after ensiling, and at the genus level, Lactiplantibacillus and Lentilactobacillus were the dominant genera in both LAB added groups, while Limosilactobacillus was the dominant genus in the CK treatment. In conclusion, the addition of LAB can improve native grass silage quality by changing bacterial community structure. LP is beneficial to improve the fermentation quality in the ensiling stage, LB is beneficial to inhibit silage deterioration in the aerobic exposure stage, and compound LAB addition is more beneficial to be applied in native grass silage.

Effects of wet brewers grains on fermentation quality and in vitro ruminal digestibility of mixed silage prepared with corn stalk, sweet potato peel and dried apple pomace in southeast China.

The effects of wet brewers grains (WBG) on fermentation quality, chemical composition and in vitro ruminal digestibility of mixed silages prepared with corn stalk, dried apple pomace and sweet potato peel were evaluated. A mixture of corn stalk, sweet potato peel and dried apple pomace (50/30/20) was ensiled with 0, 10%, 20% and 30% WBG on a fresh weight (FW) basis for 1, 3, 5, 7, 14 and 30 days respectively. The results showed that the application of WBG increased (p < 0.05) lactic acid, acetic acid and total volatile fatty acids contents, and decreased (p < 0.05) pH, dry matter, water-soluble carbohydrates content and ammonia-nitrogen/total nitrogen during ensiling. The pH in all silages was below 4.03 during ensiling. Treating with WBG increased (p < 0.05) crude protein content, and decreased (p < 0.05) neutral detergent fibre, acid detergent fibre, cellulose and hemicellulose content after 30 days of ensiling. After 72 h of incubation, cumulative gas production, potential gas production and in vitro crude protein digestibility increased (p < 0.05) with the increasing proportions of WBG. However, in vitro digestibility of dry matter and neutral detergent fibre, and metabolisable energy were similar in all silages. The 20% and 30% WBG-treated silages showed better fermentation quality and greater or higher in vitro digestibility, which were indicated by greater or higher (p < 0.05) lactic acid content, in vitro crude protein digestibility, and lower (p < 0.05) pH, ammonia-nitrogen/total nitrogen ratio as compared with the control. Therefore, ensiling agro-food by-products with at least 20% WBG were recommended for improving fermentation quality.

Study on Dynamic Fermentation of Oat Silage Assisted by Exogenous Fibrolytic Enzymes.

Based on the low content of water-soluble carbohydrate (WSC) and lactic acid bacteria (LAB) attachment in oat raw materials, we assumed that the neutral detergent fiber (NDF) content of oat can be reduced by adding cellulase or xylanase. The concentration of metabolizable sugars will be increased, which will assist the oat's bacterial community in fermentation and obtain a better quality of oat silage. After wilting the oat, it was treated as follows: (1) distributed water (CK); (2) silages inoculated with xylanase (X); and (3) silages inoculated with cellulase (C), ensiling for 3, 7, 14, 30, and 60 days. Cellulase and xylanase treatments both alter the fermentation and nutritional quality of ensiled oat, resulting in lower NDF, acid detergent fiber (ADF), cellulose, and hemicellulose contents, increased lactic acid and acetic acid contents, and a significant decrease in ensiling environment pH. The bacterial community undergoes significant changes with cellulase and xylanase treatments, with a significant increase in Lactobacillus abundance in the C_14, X_30, C_30, X_60, and C_60 treatment groups, while Weissella abundance gradually decreases with longer ensiling times. Two exogenous fibrolytic enzymes also alter the bacterial diversity of ensiled oat, with different bacterial species and abundances observed in different treatment groups. Ensiled oat treated with cellulase and xylanase experiences significant changes in its own bacterial community, particularly in the abundance of Lactobacillus. These changes result in improved fermentation and nutritional quality of oat, but the higher metabolism levels observed after 60 days of ensiling with cellulase treatment may lead to energy loss.

Influences of Growth Stage and Ensiling Time on Fermentation Characteristics, Nitrite, and Bacterial Communities during Ensiling of Alfalfa.

This study examined the impacts of growth stage and ensiling duration on the fermentation characteristics, nitrite content, and bacterial communities during the ensiling of alfalfa. Harvested alfalfa was divided into two groups: vegetative growth stage (VG) and late budding stage (LB). The fresh alfalfa underwent wilting until reaching approximately 65% moisture content, followed by natural fermentation. The experiment followed a completely randomized design, with samples collected after the wilting of alfalfa raw materials (MR) and on days 1, 3, 5, 7, 15, 30, and 60 of fermentation. The growth stage significantly influenced the chemical composition of alfalfa, with crude protein content being significantly higher in the vegetative growth stage alfalfa compared to that in the late budding stage (p < 0.05). Soluble carbohydrates, neutral detergent fiber, and acid detergent fiber content were significantly lower in the vegetative growth stage compared to the late budding stage (p < 0.05). Nitrite content, nitrate content, nitrite reductase activity, and nitrate reductase activity were all significantly higher in the vegetative growth stage compared to the late budding stage (p < 0.05). In terms of fermentation parameters, silage from the late budding stage exhibited superior characteristics compared to that from the vegetative growth stage. Compared to the alfalfa silage during the vegetative growth stage, the late budding stage group exhibited a higher lactate content and lower pH level. Notably, butyric acid was only detected in the silage from the vegetative growth stage group. Throughout the ensiling process, nitrite content, nitrate levels, nitrite reductase activity, and nitrate reductase activity decreased in both treatment groups. The dominant lactic acid bacteria differed between the two groups, with Enterococcus being predominant in vegetative growth stage alfalfa silage, and Weissella being predominant in late budding stage silage, transitioning to Lactiplantibacillus in the later stages of fermentation. On the 3rd day of silage fermentation, the vegetative growth stage group exhibited the highest abundance of Enterococcus, which subsequently decreased to its lowest level on the 15th day. Correlation analysis revealed that lactic acid bacteria, including Limosilactobacillus, Levilactobacillus, Loigolactobacillus, Pediococcus, Lactiplantibacillus, and Weissella, played a key role in nitrite and nitrate degradation in alfalfa silage. The presence of nitrite may be linked to Erwinia, unclassified_o__Enterobacterales, Pantoea, Exiguobacterium, Enterobacter, and Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium.

Effects of Isolated LAB on Chemical Composition, Fermentation Quality and Bacterial Community of Stipa grandis Silage.

This study aimed to screen and identify lactic acid bacteria (LAB) strains from the Stipa grandis and naturally fermented silage, and assess their effects on the silage quality and bacterial community of Stipa grandis after 60 days of the fermentation process. A total of 38 LAB were isolated, and strains ZX301 and YX34 were identified as Lactiplantibacillus plantarum and Pediococcus pentosaceus using 16S rRNA sequences; they can normally grow at 10−30 °C, with a tolerance of pH and NaCl from 3.5 to 8.0 and 3 to 6.5%, respectively. Subsequently, the two isolated LAB and one commercial additive (Lactiplantibacillus plantarum) were added to Stipa grandis for ensiling for 60 days and recorded as the ZX301, YX34, and P treatments. The addition of LAB was added at 1 × 105 colony-forming unit/g of fresh weight, and the same amount of distilled water was sprayed to serve as a control treatment (CK). Compared to the CK treatment, the ZX301 and YX34 treatments exhibited a positive effect on pH reduction. The water-soluble carbohydrate content was significantly (p < 0.05) increased in ZX301, YX34, and P treatments than in CK treatment. At the genus level, the bacterial community in Stipa grandis silage involves a shift from Pantoea to Lactiplantibacillus. Compared to the CK treatment, the ZX301, YX34, and P treatments significantly (p < 0.05) increase the abundance of Pediococcus and Lactiplantibacillus, respectively. Consequently, the results indicated that the addition of LAB reconstructed microbiota and influenced silage quality. The strain ZX301 could improve the ensiling performance in Stipa grandis silage.

Fermentation quality, bacterial community, and predicted functional profiles in silage prepared with alfalfa, perennial ryegrass and their mixture in the karst region.

There is little information regarding the dynamics of fermentation products and the bacterial community in silage prepared with alfalfa (MS), perennial ryegrass (LP), and their mixture in the karst region. In this study, we explored the effects of combining MS with LP in different ratios (100% MS, 70% MS + 30% LP, 50% MS + 50% LP, 30% MS + 70% LP and 100% LP; fresh matter basis) on silage chemical composition, fermentation quality, bacterial communities and predicted functions during the ensiling process. Each treatment was prepared in triplicate and stored at room temperature (22-25°C) for 7, 15, and 45 days. The dry matter (DM) and water-soluble carbohydrate content of the silages increased as the LP proportion in the mixed silage increased; at 45 days, the 70% MS + 30% LP, 50% MS + 50% LP and 30% MS + 70% LP silages contained higher (p < 0.05) CP content than the 100% MS and 100% LP silages. The 30% MS + 70% LP and 100% LP silages exhibited lower (p < 0.05) pH and higher (p < 0.05) LA content than the other silages; at 45 days, none of the silages contained PA or BA. As fermentation proceeded, the abundance of harmful (Enterobacteriaceae and Sphingomonas) and beneficial (Lentilactobacillus, Lactiplantibacillus, Secundilactobacillus, and Levilactobacillus) microorganisms decreased and increased, respectively, as the LP proportion in the mixed silage increased. The predicted functional distribution of microbial communities and metabolic pathways revealed that the 30% MS + 70% LP and 100% LP silages had a stronger capacity for fermentation and a weaker capacity for nitrate reduction than the other silages. Moreover, as the fermentation proceeded, the 30% MS + 70% LP and 100% LP treatments enhanced the functions of "Metabolism," "Genetic information processing" and "Organismal systems" at level 1, the functions of "Amino acid metabolism" and "Nucleotide metabolism" at level 2, and the functions of "Metabolic pathways," "Biosynthesis of secondary metabolites," "Biosynthesis of antibiotics" and "Purine metabolism" at level 3. Thus, adding LP could improve the fermentation quality of MS silage by changing the composition and metabolic function of microbes; furthermore, ensiling 30% alfalfa with 70% ryegrass can produce high-quality silage in the karst region.

Evaluating the fermentation characteristics, bacterial community, and predicted functional profiles of native grass ensiled with different additives.

Bioaugmentation of native grass ensiling with Lactobacillus plantarum or Lactobacillus buchneri or Pediococcus pentosaceus on the ensiling performance and bacterial community was investigated after 30 days of the fermentation process. The native grass was inoculated with distilled water, Lactobacillus plantarum, Lactobacillus plantarum, and Lactobacillus buchneri, and Lactobacillus plantarum, Lactobacillus buchneri, and Pediococcus pentosaceus as the CON treatment, T1 treatment, T2 treatment, and T3 treatment, respectively. The addition of lactic acid bacteria was added at a total of 1 × 106 colony-forming unit/g of fresh weight. As expected, the markedly (p < 0.05) lower water-soluble carbohydrate content was tested in the T2 and T3 treatments compared to the CON and T1 treatments. Compared to the CON and T1 treatment, significantly (p < 0.05) higher crude protein content, and lower acid detergent fiber and neutral detergent fiber contents were found in the T2 and T3 treatments. Compared to the CON treatment, the pH significantly (p < 0.05) decreased in the lactic acid bacteria (LAB) inoculated silage, and the lowest pH was measured in the T3 treatment. Similarly, significantly higher lactic acid and acetic acid contents were also found in the T3 treatment compared to those in other treatments. After 30 days of ensiling, the Shannon and Chao1 indexes in silages decreased compared to that in the fresh materials (FMs). The principal coordinate analysis indicated that both FM and silage were distinctly separated in each treatment with no interactions on the confidence ellipse (R = 0.8933, p = 0.001). At the phylum level, the dominant phylum was shifted from Proteobacteria to Firmicutes after the fermentation process. Interestingly, Weissella dominated the fermentation in the CON treatment and Lactobacillus dominated the fermentation in all inoculated LAB silages at the genus level. Results of functional prediction analyses showed that the metabolism of amino acid, cofactors, and vitamins, and membrane transport was reduced, while the metabolism of nucleotide and majority carbohydrates was increased after ensiling. The complex LAB (Lactobacillus plantarum, Lactobacillus buchneri, and Pediococcus pentosaceus) exhibited the potential possibility to decrease pH and enhance the relative abundance of LAB in response to obtaining high-quality silage by the synergistic effects. These results suggested that the complex LAB could improve the ensiling performance of native grass silage, and lay a theoretical basis for inoculant application in native grass.