Microbial community structure, co-occurrence network and fermentation characteristics of woody plant silage.

BACKGROUND: Feed shortage is a factor restricting animal production in the tropics, therefore how to use natural woody plant resources as animal feed is an important strategy. RESULTS: Under the dual stress of an anaerobic and acidic environment, the microbial response during the fermentation of paper mulberry (PM) silage was found to be sensitive. The Gram-negative bacteria and mould died, and the dominant microbial community rapidly shifted to Gram-positive bacteria, resulting in a large reduction in microbial diversity and abundance. Exogenous bran additives interfered with the stress effects of the woody silage environment. Wheat bran (WB) accelerated the response of microorganisms to the anaerobic stress, and lactic acid bacteria became the dominant microbial community, thereby enhancing the lactic acid fermentation of silage, affecting the metabolic pathways of microorganisms, and improving the flavour and quality of the silage. Addition of rice bran made Enterobacter and Clostridium species quickly respond to the stress of the silage environment and become the predominant bacterial groups. In particular, anaerobic and spore-forming Clostridium species showed a strong tolerance to the silage environment, leading to butyric acid fermentation and protein degradation of the silage, and reducing its fermentation quality. CONCLUSION: The PacBio single-molecule real-time (SMRT) sequencing technology accurately revealed the microbial co-occurrence network and fermentation mechanism of silage. Our results indicate that PM can be used in combination with WB to prepare high-quality silage for animal production. © 2021 Society of Chemical Industry.

Dynamic changes and characterization of the protein and carbohydrate fractions of native grass grown in Inner Mongolia during ensiling and the aerobic stage.

OBJECTIVE: To improve the utility of native grass resources as feed in China, we investigated the dynamics of protein and carbohydrate fractions among Inner Mongolian native grasses, during ensiling and the aerobic stage, using the Cornell Net Carbohydrate and Protein System. METHODS: Silages were prepared without or with lactic acid bacteria (LAB) inoculant. We analyzed the protein and carbohydrate fractions and fermentation quality of silages at 0, 5, 15, 20, 30, and 60 d of ensiling, and the stability at 0.5, 2, 5, and 10 d during the aerobic stage. RESULTS: Inner Mongolian native grass contained 10.8% crude protein (CP) and 3.6% water-soluble carbohydrates (WSC) on a dry matter basis. During ensiling, pH and CP and WSC content decreased (p<0.05), whereas lactic acid and ammonia nitrogen (N) content increased (p<0.05). Non-protein N (PA) content increased significantly, whereas rapidly degraded true protein (PB1), intermediately degraded true protein (PB2), total carbohydrate (CHO), sugars (CA), starch (CB1), and degradable cell wall carbohydrate (CB2) content decreased during ensiling (p<0.05). At 30 d of ensiling, control and LAB-treated silages were well preserved and had lower pH (<4.2) and ammonia-N content (<0.4 g/kg of fresh matter [FM]) and higher lactic acid content (>1.0% of FM). During the aerobic stage, CP, extract ether, WSC, lactic acid, acetic acid, PB1, PB2, true protein degraded slowly (PB3), CHO, CA, CB1, and CB2 content decreased significantly in all silages, whereas pH, ammonia-N, PA, and bound true protein (PC) content increased significantly. CONCLUSION: Control and LAB-treated silages produced similar results in terms of fermentation quality, aerobic stability, and protein and carbohydrate fractions. Inner Mongolian native grass produced good silage, nutrients were preserved during ensiling and protein and carbohydrate losses largely occurred during the aerobic stage.

Community of natural lactic acid bacteria and silage fermentation of corn stover and sugarcane tops in Africa.

OBJECTIVE: To effectively utilize crop by-product resources to address the shortage of animal feed during the dry season in Africa, the community of natural lactic acid bacteria (LAB) of corn stover and sugarcane tops and fermentation characteristics of silage were studied in Mozambique. METHODS: Corn stover and sugarcane tops were obtained from agricultural field in Mozambique. Silage was prepared with LAB inoculant and cellulase enzyme and their fermentation quality and microbial population were analyzed. RESULTS: Aerobic bacteria were the dominant population with 107 colony-forming unit/g of fresh matter in both crops prior to ensiling, while 104 to 107 LAB became the dominant bacteria during ensiling. Lactobacillus plantarum was more than 76.30% of total isolates which dominated silage fermentation in the LAB-treated sugarcane top silages or all corn stover silages. Fresh corn stover and sugarcane tops contain 65.05% to 76.10% neutral detergent fiber (NDF) and 6.52% to 6.77% crude protein (CP) on a dry matter basis, and these nutrients did not change greatly during ensiling. Corn stover exhibits higher LAB counts and watersoluble carbohydrates content than sugarcane top, which are naturally suited for ensiling. Meanwhile, sugarcane tops require LAB or cellulase additives for high quality of silage making. CONCLUSION: This study confirms that both crop by-products contain certain nutrients of CP and NDF that could be well-preserved in silage, and that they are potential roughage resources that could cover livestock feed shortages during the dry season in Africa.

Influence of microbial additive on microbial populations, ensiling characteristics, and spoilage loss of delayed sealing silage of Napier grass.

OBJECTIVE: To measure whether a microbial additive could effectively improve the fermentation quality of delayed-sealing (DS) silage, we studied the effects of inoculants of lactic acid bacteria (LAB) and cellulase enzyme on microbial populations, ensiling characteristics, and spoilage loss of DS silage of Napier grass in Africa. METHODS: Quick-sealing (QS) and DS silages were prepared with and without LAB (Lactobacillus plantarum) inoculant, cellulase enzymes, and their combination. The QS material was directly chopped and packed into a bunker silo. The DS material was packed into the silo with a delay of 24 h from harvest. RESULTS: In the QS silage, LAB was dominant in the microbial population and produced large amounts of lactic acid. When the silage was treated with LAB and cellulase, the fermentation quality was improved. In the DS silage, aerobic bacteria and yeasts were the dominant microbes and all the silages were of poor quality. The yeast and mold counts in the DS silage were high, and they increased rapidly during aerobic exposure. As a result, the DS silages spoiled faster than the QS silages upon aerobic exposure. CONCLUSION: DS results in poor silage fermentation and aerobic deterioration. The microbial additive improved QS silage fermentation but was not effective for DS silage.

Microbial network and fermentation modulation of Napier grass and sugarcane top silage in southern Africa.

IMPORTANCE: Feed shortage in the tropics is a major constraint to the production of livestock products such as milk and meat. In order to effectively utilize of local feed resources, the selected lactic acid bacteria (LAB) strain was used to prepare Napier grass and sugarcane top silage. The results showed that the two silages inoculated with LAB formed a co-occurrence microbial network dominated by Lactiplantibacillus during the fermentation process, regulated the microbial community structure and metabolic pathways, and improved the silage fermentation quality. This is of great significance for alleviating feed shortage and promoting sustainable production of livestock.

Cleaner anaerobic fermentation and greenhouse gas reduction of crop straw.

Rice anaerobic fermentation is a significant source of greenhouse gas (GHG) emissions, and in order to efficiently utilize crop residue resources to reduce GHG emissions, rice straw anaerobic fermentation was regulated using lactic acid bacteria (LAB) inoculants (FG1 and TH14), grass medium (GM) to culture LAB, and Acremonim cellulolyticus (AC). Microbial community, GHG emission, dry matter (DM) loss, and anaerobic fermentation were analyzed using PacBio single-molecule real-time and anaerobic fermentation system. The epiphytic microbial diversity of fresh rice straw was extremely rich and contained certain nutrients and minerals. During ensiling, large amounts of GHG such as carbon dioxide are produced due to plant respiration, enzymatic hydrolysis reactions, and proliferation of aerobic bacteria, resulting in energy and DM loss. Addition of FG1, TH14, and AC alone improved anaerobic fermentation by decreasing pH and ammonia nitrogen content (P < 0.05) and increased lactic acid content (P < 0.05) when compared to the control, and GM showed the same additive effect as LAB inoculants. Microbial additives formed a co-occurrence microbial network system dominated by LAB, enhanced the biosynthesis of secondary metabolites, diversified the microbial metabolic environment and carbohydrate metabolic pathways, weakened the amino acid metabolic pathways, and made the anaerobic fermentation cleaner. This study is of great significance for the effective utilization of crop straw resources, the promotion of sustainable livestock production, and the reduction of GHG emissions.IMPORTANCETo effectively utilize crop by-product resources, we applied microbial additives to silage fermentation of fresh rice straw. Fresh rice straw is extremely rich in microbial diversity, which was significantly reduced after silage fermentation, and its nutrients were well preserved. Silage fermentation was improved by microbial additives, where the combination of cellulase and lactic acid bacteria acted as enzyme-bacteria synergists to promote lactic acid fermentation and inhibit the proliferation of harmful bacteria, such as protein degradation and gas production, thereby reducing GHG emissions and DM losses. The microbial additives accelerated the formation of a symbiotic microbial network system dominated by lactic acid bacteria, which regulated silage fermentation and improved microbial metabolic pathways for carbohydrates and amino acids, as well as biosynthesis of secondary metabolites.

Cellulase-lactic acid bacteria synergy action regulates silage fermentation of woody plant.

BACKGROUND: Feed shortage is an important factor limiting livestock production in the world. To effectively utilize natural woody plant resources, we used wilting and microbial additives to prepare an anaerobic fermentation feed of mulberry, and used PacBio single-molecule real-time (SMRT) sequencing technology to analyse the "enzyme-bacteria synergy" and fermentation mechanism. RESULTS: The fresh branches and leaves of mulberry have high levels of moisture and nutrients, and also contain a diverse range of epiphytic microorganisms. After ensiling, the microbial diversity decreased markedly, and the dominant bacteria rapidly shifted from Gram-negative Proteobacteria to Gram-positive Firmicutes. Lactic acid bacteria (LAB) emerged as the dominant microbial population, resulting in increased in the proportion of the carbohydrate metabolism and decreased in the proportion of the amino acid and "global and overview map" (GOM) metabolism categories. The combination of cellulase and LAB exhibited a synergistic effect, through which cellulases such as glycanase, pectinase, and carboxymethyl cellulase decomposed cellulose and hemicellulose into sugars. LAB converted these sugars into lactic acid through the glycolytic pathway, thereby improving the microbial community structure, metabolism and fermentation quality of mulberry silage. The GOM, carbohydrate metabolism, and amino acid metabolism were the main microbial metabolic categories during ensiling. The presence of LAB had an important effect on the microbial community and metabolic pathways during silage fermentation. A "co-occurrence microbial network" formed with LAB, effectively inhibiting the growth of harmful microorganisms, and dominating the anaerobic fermentation process. CONCLUSIONS: In summary, PacBio SMRT was used to accurately analyse the microbial network information and regulatory mechanism of anaerobic fermentation, which provided a scientific basis for the study of woody silage fermentation theory. This study reveals for the first time the main principle of the enzyme-bacteria synergy in a woody silage fermentation system, which provides technical support for the development and utilization of woody feed resources, and achieves sustainable livestock production.

Silage preparation and sustainable livestock production of natural woody plant.

As the global population increases and the economy grows rapidly, the demand for livestock products such as meat, egg and milk continue to increase. The shortage of feed in livestock production is a worldwide problem restricting the development of the animal industry. Natural woody plants are widely distributed and have a huge biomass yield. The fresh leaves and branches of some woody plants are rich in nutrients such as proteins, amino acids, vitamins and minerals and can be used to produce storage feed such as silage for livestock. Therefore, the development and utilization of natural woody plants for clean fermented feed is important for the sustainable production of livestock product. This paper presents a comprehensive review of the research progress, current status and development prospects of forageable natural woody plant feed resources. The nutritional composition and uses of natural woody plants, the main factors affecting the fermentation of woody plant silage and the interaction mechanism between microbial co-occurrence network and secondary metabolite are reviewed. Various preparation technologies for clean fermentation of woody plant silage were summarized comprehensively, which provided a sustainable production mode for improving the production efficiency of livestock and producing high-quality livestock product. Therefore, woody plants play an increasingly important role as a potential natural feed resource in alleviating feed shortage and promoting sustainable development of livestock product.

Using PacBio SMRT Sequencing Technology and Metabolomics to Explore the Microbiota-Metabolome Interaction Related to Silage Fermentation of Woody Plant.

Silage fermentation is a dynamic process involving the succession of microbial communities and changes in metabolites. Fresh branched and leaves of paper mulberry were used to prepared silage. Crop by-products including corn bran, rice bran, and wheat bran were used as exogenous additives. Pacific Biosciences single-molecule real-time (SMRT) sequencing technology and metabolomics are used to explore the interaction mechanism of microbial structure and metabolites during woody silage fermentation and to verify the principle that exogenous additives can modulate silage fermentation. Under the dual stress of anaerobic and acidic environment of silage fermentation, the microbial community changed from Gram-negative to Gram-positive bacteria, and a large amount of lactic acid and volatile fatty acid were produced, which lowered the pH value and caused the rapid death of aerobic bacteria with thin cell walls. The exogenous additives of corn bran and wheat bran accelerated the dynamic succession of lactic acid bacteria as the dominant microbial community in silage fermentation, increased the metabolic pathways of lactic acid, unsaturated fatty acids, citric acid, L-malic acid and other flavoring agents, and inhibited the growth of Clostridium and Enterobacter, thereby improving the flavor and quality of the silage. However, because rice bran contained butyric acid spore bacteria, it can multiply in an anaerobic environment, led to butyric acid fermentation, and promoted protein degradation and ammonia nitrogen production, thereby reduced the fermentation quality of woody silage. The results showed that during the silage fermentation process, the microbial community and the metabolome can interact, and exogenous additives can affect the fermentation quality of silage. SMRT sequencing technology and untargeted metabolomics revealed the microbiota-metabolome interaction during silage fermentation. Changes in the structure of the microbial community can affect the metabolic pathways, and the final metabolites can inhibit the growth of microorganisms that are not conducive to silage fermentation. Exogenous carbohydrate additives can change the fermentation substrate and affect microbial community structure, thus modulate the silage fermentation.

Microbial Co-occurrence Network and Fermentation Information of Natural Woody-Plant Silage Prepared With Grass and Crop By-Product in Southern Africa.

To facilitate the use of woody plant (WP) as a natural biomass resource to address the shortage of feed for ruminants in the tropics, we use PacBio SMRT sequencing to explore the microbial co-occurrence network and silage fermentation of gliricidia and leucaena prepared with Napier grass (NG) and corn stover (CS) in Southern Africa. Based on dry matter, the crude protein contents of WP are as high as 25%. Compared with NG, the addition of CS speed up the dynamic succession of microorganisms in the silage fermentation process from Gram-negative bacteria to Gram-positive bacteria, and promoted Lactiplantibacillus plantarum to become the dominant community and enhanced the metabolic pathways of lactic acid and citric acid, thus improved the fermentation flavour and quality of WP silage. WP can be mixed with CS to make high-quality silage, which can alleviate the shortage of feed and promote local animal production.

Chemical Modification, Characterization, and Activity Changes of Land Plant Polysaccharides: A Review.

Plant polysaccharides are widely found in nature and have a variety of biological activities, including immunomodulatory, antioxidative, and antitumoral. Due to their low toxicity and easy absorption, they are widely used in the health food and pharmaceutical industries. However, low activity hinders the wide application. Chemical modification is an important method to improve plant polysaccharides' physical and chemical properties. Through chemical modification, the antioxidant and immunomodulatory abilities of polysaccharides were significantly improved. Some polysaccharides with poor water solubility also significantly improved their water solubility after modification. Chemical modification of plant polysaccharides has become an important research direction. Research on the modification of plant polysaccharides is currently increasing, but a review of the various modification studies is absent. This paper reviews the research progress of chemical modification (sulfation, phosphorylation, acetylation, selenization, and carboxymethylation modification) of land plant polysaccharides (excluding marine plant polysaccharides and fungi plant polysaccharides) during the period of January 2012-June 2022, including the preparation, characterization, and biological activity of modified polysaccharides. This study will provide a basis for the deep application of land plant polysaccharides in food, nutraceuticals, and pharmaceuticals.

Chemical composition, characteristics concerned with fermentative quality and microbial population of ensiled pearl millet and sorghum stover in semi-arid West Africa.

To effectively utilize crop by-product resources for ruminant feed in semi-arid West Africa, we studied the chemical composition, characteristics concerned with fermentative quality, and microbial population of ensiled pearl millet stover (PMS) and sorghum stover (SS) in Mozambique. After panicle harvest, the PMS and SS were exposed in the field for 7, 21, 35, 49, 63, 77, 91, and 120 days under natural weather conditions. The fresh stover silages were prepared and stored for 120 days. With increased exposure time, the dry matter, neutral detergent fiber, acid detergent lignin, and neutral detergent insoluble nitrogen contents increased, whereas the crude protein, ether extract, gross energy, digestible energy, metabolizable energy, and true protein contents decreased. After 120 days of field exposure, aerobic bacteria dominated both stovers, and lactic acid bacteria (LAB) decreased to below detectable levels. After 120 days of ensiling, LAB dominated the silage of both crops, while the harmful microorganisms as aerobic bacteria, coliform bacteria, yeast, and mold were reduced or below detectable levels. Both silages did not produce more lactic acid to reduce the pH value, but they preserved nutrients well during ensiling. Therefore, PMS and SS can be prepared as silage for ruminant feed in semi-arid West Africa.

Ensiling characteristics of total mixed ration prepared with local feed resources in Mozambique and their effects on nutrition value and milk production in Jersey dairy cattle.

To effectively utilize total mixed ration (TMR) prepared with locally available feed resources, we studied the nutritional value and milk production in Jersey dairy cattle fed a local general diet (LGD) and fermented TMR in Mozambique. Ten head of Jersey dairy cattle with 337 ± 19.8 kg body weight, aged 3-4 years in mid location were used in this study. The LGD diet was designed following the general feeding method of local smallholding farms; it contained native grass, Napier grass, wheat bran, and mineral-vitamin mix. Fermented TMR was prepared using Napier grass, corn bran, wheat bran, formula feed, and mineral-vitamin mix. Fermented TMR was preserved as good quality, with a relatively low pH and high lactic acid content. Compared to LGD, fermented TMR significantly improved the dry matter intake and digestibility in dairy cattle. Milk yield was significantly higher in dairy cattle fed with TMR than in those fed with LGD, by 3.75 L/d; milk quality was not significantly different between treatments. The results confirmed that LGD had a low dry matter intake and milk yield, and fermented TMR prepared with local feed resources can attain good quality and improve milk yield in dairy cattle in Mozambique.

Growth performance, apparent digestibility, and N balance in Mongolian lambs and hoggs fed diets supplemented with a Chinese traditional herbal medicine complex.

We evaluated the effects of dietary supplementation with Chinese traditional herbal medicine (CTHM) on the growth performance, apparent digestibility, and nitrogen balance in Mongolian lambs and hoggs. The CTHM used as a dietary supplement consisted of 30% pine needles, 20% mugwort, 40% garlic, and 10% Astragalus mongholicus on a dry matter (DM) basis. The basal diet was prepared from corn stover, corn grain, wheat bran, rapeseed meal, shell meal, dicalcium phosphate, salt, vitamin premix, and mineral premix. In total, 16 Mongolian lambs (20.75 kg initial body weight [BW]) and 16 hoggs (33.81 kg initial BW) were allocated randomly to one of two treatments, the basal diet without and with CTHM, with the group receiving CTHM fed 98.5% basal diet + 1.5% CTHM complex on a DM basis. Lamb and hogg growth trials showed that CTHM supplementation improved the final live weight, gain, and feed conversion ratio (p < 0.01). A digestibility trial showed that the diet with CTHM improved the digestibility of DM, organic matter (OM), crude protein (CP), and neutral detergent fiber in lambs and hoggs. Finally, CTHM supplementation decreased (p < 0.0001) fecal and urinary nitrogen and increased (p = 0.0004) nitrogen (N) retention. Overall, the addition of CTHM to the diets of lambs and hoggs has beneficial effects on growth.