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BackgroundPaper mulberry is a promising alternative fodder source due to its high protein and the abundance of active components. However, paper mulberry often faces susceptibility to contamination during silage fermentation, and there is a need to improve the quality of silage fermentation of paper mulberry through exotic additives. Sea buckthorn pomace (BP) is a feed additive containing antimicrobial and antioxidant substances that help to enhance silage fermentation. Therefore, the objective of this study was to evaluate the effects of BP and Lactiplantibacillus as additives on silage fermentation and bacterial community of paper mulberry.ResultsThe results showed that BP and Lactiplantibacillus significantly reduced the pH and ammonium nitrogen content of paper mulberry silage (P < 0.05) and significantly increased the content of lactic acid and acetic acid (P < 0.05), resulting in more residual water-soluble carbohydrate and crude protein contents and less fiber content relative to the control. The key microorganisms in paper mulberry silage fermentation are Lactiplantibacillus pentosus and Weissella cibaria. Among these, Lactiplantibacillus favored a rapid increase in Lactiplantibacillus pentosus abundance during the pre-silage fermentation period, whereas BP favored the promotion of Lactiplantibacillus pentosus growth, resulting in higher contents of lactic and acetic acid than those of the control.ConclusionsSimultaneously adding Lactiplantibacillus and BP can effectively improve the quality of paper mulberry silage and increase the abundance of beneficial microorganisms in paper mulberry silage.

BackgroundIn this study, we aimed to address the low utilization of straw and poor fermentation quality of paper mulberry silage (under natural fermentation conditions). Straw was combined with paper mulberry for ensiling, and the fermentation characteristics, bacterial community, and metabolite composition of the mixed straw and paper mulberry silage were investigated. Four treatment groups were established: corn-straw treatment 2 (3:7 ratio of corn straw to paper mulberry), corn-straw treatment 3 (5:5 ratio of corn straw to paper mulberry), wheat-straw treatment 2 (3:7 ratio of wheat straw to paper mulberry), wheat-straw treatment 3 (5:5 ratio of wheat straw to paper mulberry), and a control group (ensiling of paper mulberry alone).ResultsThe control group demonstrated the highest pH and ammonia (AN) and acetic acid (AA) content compared with all the treatment groups. Corn-straw treatment 2 had the highest lactic acid content (54.70 g/kg dry weight) compared with the control and other treatment groups. The relative abundance of Enterobacter (7.085%) was the lowest in the control than in the other treatment groups (p

There is an increasing worldwide demand for biogas. Anaerobic co-digestion involves the treatment of different substrates with the aim of improving the production of biogas and the stability of the process. This study evaluates how methane production is affected by the co-digestion of pig and dairy manure with grass silage and pulp and paper mill sludge and assesses whether methane production is affected by factors other than nutrient deficiency, low buffering capacity, inadequate dilution, and an insufficient activity and amount of microorganism culture. Anaerobic digestion was performed in batch reactors under mesophilic conditions for 20 days. The season of grass silage and manure collection proved to be an important factor affecting methane production. Spring grass silage produced a maximum of 250 mL/VSadded and spring manure 150 mL/VSadded, whereas autumn grass silage produced at most 140 ml/VSadded and autumn manure 45 mL/VSadded. The pulp mill sludge used is comprised of both primary and secondary sludge and produced at most 50 mL/VSadded regardless of season; this substrate benefitted most from co-digestion.

To investigate the effects of different organic acid additives and their concentrations on the fermentation quality and bacterial community of paper mulberry silage, paper mulberry was left untreated (control) or was treated with ethylenediaminetetraacetic acid (EDTA), propionic acid (PA) or citric acid (CA), the amount of each additive was 2 g.kg−1 FM, 5 g.kg−1 FM and g.kg−1 FM. All groups were ensiled for 3, 7, 15, 30 and 60 days. Compared to the control, adding EDTA reduced protein breakdown, preserved more water-soluble carbohydrates of the silages (WSCs, 24.74 g.kg−1 DM), and high concentrations of EDTA inhibited the activity of undesirable microorganisms. Adding PA increased the abundance of Lactiplantibacillus and decreased the abundance of Enterococcus, and it caused a rapid decrease in the pH of the silage at an early stage (from 6.50 to 5.31) while altering the microbiota, and low concentrations of PA resulted in high LA (66.22 g.kg−1 DM) concentration and low PA (9.92 g.kg−1 DM) concentration at 60 days of ensiling. Different concentrations of additives altered the microbial community of paper mulberry to different degrees. High concentrations of PA and CA can increase the abundance of Lactiplantibacillus. High concentrations of CA resulted in a rapid decrease in silage pH at an early stage and higher WSC concentration. These results suggest that EDTA, PA and CA can be used as additives to improve the quality of paper mulberry silage.

Paper mulberry (Broussonetia papyrifera L., PM) is being used as a new type of animal protein feed to address the feed crisis. To investigate the effect of additives on the chemical composition, fermentation quality, and bacterial community of PM silage (at room temperature, 25°), paper mulberry was fermented with formic acid (FA), Amomum villosum essential oil (AVEO) and lactic acid bacteria (LAB) inoculant treatments. The results showed that fresh PM had a low water-soluble carbohydrate (WSC) content and large amounts of unclassified bacteria. Compared with the CK and LAB treatments, the FA and AVEO treatments significantly (P < 0.05) decreased the pH and increased the lactic acid content of PM silage after 60 days of ensiling. In the AVEO-treated silages the abundance of Lactococcus in the early stage of ensiling increased by 14.09%, the abundances of Levilactobacillus and Lentilactobacillus in the late stage of ensiling increased by 58.34 and 91.12%, respectively, and the abundance of Stenotrophomonas decreased by 94.71%, resulting in improved PM silage quality. These results confirmed that AVEO could potentially be developed as a new additive for improving the fermentation quality of silage.

This study aimed to isolate, characterize, and identify lactic acid bacteria (LAB) strains from various sources and evaluate their effects on the nutritional quality, fermentation characteristics, and microbial compositions of paper mulberry (PM) after 60 days of ensiling. Forty-nine LAB strains were isolated from Phalaris arundinacea silage, pickle, and fresh PM leaves; three of these strains (Lactiplantibacillus plantarum, YC1; Levilactobacillus brevis, PC3; and Lactiplantibacillus plantarum, BP17) and one commercial inoculant Gaofuji (GFJ) were subsequently used. Compared with other treatments, PC3 and BP17 increased (P < 0.05) the LAB count and crude protein content and decreased (P < 0.05) the molds and coliform bacteria counts, pH, and ammonia-N content of PM silages. BP17 and PC3 increased the relative Lactiplantibacillus abundance and decreased that of Lelliottia and Cladosporium, improving PM silage quality. Therefore, PC3 and BP17 can improve the fermentation quality of PM silage and could be used as silage starter cultures.

Lactic acid bacteria occupy an important position in silage microorganisms, and the effects of exogenous lactic acid bacteria on silage quality have been widely studied. Microbial metabolism has been proved as an indicator of substrate utilization by microorganisms. Paper mulberry is rich in free carbohydrate, amino acids, and other components, with the potential to be decomposed and utilized. In this study, changes in the microbial metabolism characteristics of paper mulberry silage with Lactiplantibacillus plantarum (LP) and Lentilactobacillus buchneri (LB) were studied along with a control (CK) using BIOLOG ECO microplates. The results showed that average well-color development (AWCD), Shannon diversity, Shannon evenness, and Simpson diversity exhibited significant temporal trends. LB and LP responded differently in the early ensiling phase, and the AWCD of LB was higher than LP at 7 days. Principal component analysis revealed that CK, LB, and LP samples initially clustered at 3 days and then moved into another similar cluster after 15 days. Overall, the microplates methodology applied in this study offers important advantages, not least in terms of accuracy.

There are few studies on the application of lactic acid bacteria in the reduction of anti-nutrient factors in paper mulberry silage. This study aimed to investigate the effects of different lactic acid bacteria on the fermentation quality and the amount of anti-nutritional factors in paper mulberry silage. Two strains of Lactobacillus plantarum (GX, isolated from paper mulberry silage; GZ, provided by Sichuan Gaofuji Biotechnology Co. Ltd.) were added as silage additives. On days 7, 15, 30 and 60 of the ensiling process, the fermentation quality, and the amount of anti-nutritional factors were measured. Compared with the control group, inoculation with Lactobacillus plantarum could rapidly reduce pH values, leading to lower NH3-N/TN. Besides, it also significantly increased the lactic acid content (p < 0.05). The two strains of L. plantarum significantly reduced the content of hydrolysed tannin, condensed tannin, total tannin, oxalic acid, phytic acid and saponin (p < 0.05). Overall, this study found that the addition of lactic acid bacteria could significantly improve the fermentation quality of paper mulberry and reduce the amount of anti-nutrient factors (p < 0.05).

Simple Summary: Paper mulberry (PM) is an unconventional protein feed material, and silage is its main processing method. The present study aimed to investigate the health benefits and meat quality of supplementing Yangzhou geese with paper mulberry silage. Results indicated that paper mulberry silage supplementation had a promotional effect on the growth of Yangzhou geese, the sensory quality of the breast muscles improved, and the nutritional quality enhanced. Untargeted metabolomics analysis demonstrated that PM treatment enhanced guanidinoacetic acid levels in breast muscles and facilitated the metabolism of amino acids and the role of substances in lipid antioxidant pathways. Consequently, paper mulberry could be considered to be a novel protein feed option capable for geese. There have been few investigations into the health benefits and meat quality of supplementing Yangzhou geese with paper mulberry silage. One hundred and twenty 28-day-old Yangzhou geese were selected for the experiment and randomly divided into two groups: a control group (CON) and a paper mulberry silage group (PM), with six replicates in each group. The experiment lasted for a total of 6 weeks. The experiment found that compared with CON, PM had a promoting effect on the average daily weight gain of Yangzhou geese (p = 0.056). Sensory and nutritional analysis of breast muscles revealed a decrease in a* value (p < 0.05) and an increase in protein content (p < 0.05) following PM treatment. Through untargeted metabolomics analysis of breast muscle samples, it was found that 11 different metabolites, including guanidinoacetic acid and other substances, had a positive effect on amino acid metabolism and lipid antioxidant pathways of PM treatment. Overall, the strategy of feeding Yangzhou geese with paper mulberry silage is feasible, which can improve the sensory quality and nutritional value of goose meat. The experiment provides basic data for the application form of goose breeding, so exploring the impact of substances within paper mulberry on goose meat should be focused on in the future. [ABSTRACT FROM AUTHOR]

In this study, the influence of two lactic acid bacteria (LAB) strains [Lactobacillus rhamnosus BDy (LR-BDy) and Lactobacillus buchneri TSy (LB-TSy)] selected from Southwest China on the fermentation characteristics and in vitro gas production of Broussonetia papyrifera (paper mulberry) silage were experimentally explored. The experimental groups were a control group (C), an LB-TSy treatment (LB), an LR-BDy treatment (LR), and an LR-BDy + LB-TSy hybrid group (LR × LB). After the LAB were added, the pH value of paper mulberry silage significantly declined (p < 0.05), and the crude protein content was effectively preserved (p < 0.05). However, no significant changes were found in the levels of neutral detergent fiber, acid detergent fiber, and crude ash (p > 0.05). The lactic acid content in paper mulberry silage was evidently increased (p < 0.05). The in vitro gas production in the LR at 36, 48, and 72 h were markedly higher than that in the other treatments (p < 0.05). Owing to the addition of LAB, the microbial diversity in paper mulberry silage was reduced, while the relative bacterial abundance of Lactobacillus was enhanced. Hence, the addition of LAB selected from the warm and humid region in Southwest China can improve the quality of paper mulberry silage and elevate its feeding value in this region.

Paper mulberry (Broussonetia papyrifera) is widely ensiled to feed sheep in southwestern China, as unconventional woody forage. Feeding lambs with paper mulberry silage (PMS) may improve certain feeding characteristics, thereby affecting the growth performance and meat quality. The aim of this study is to investigate the effects of four diets of PMS on growth performance, rumen microbial composition, and muscle fatty acids profile in Hu lambs. The results showed that 30% and 40% PMS increased the dry matter intake and average daily gain of Hu lambs compared to the control group. PMS30 and PMS40 increased the content of C24:1, and PMS40 increased the content of C20:5n-3. The content of microbial protein (MCP) was higher in PMS40 than in others, but PMS30 and PMS40 reduced the total volatile fatty acid in rumen. PMS30 significantly increased the ratio of acetic acid to propionic acid. The abundance of ruminal Christensenellaceae_R-7_group and norank_f_Eubacterium_coprostanoligenes_group was significantly higher in PMS30 and PMS40 groups. Moreover, Christensenellaceae_R-7_group had a significant positive correlation with n3-polyunsaturated fatty acid. PMS40 might lead to a relatively high content of unsaturated fatty acids in longissimus dorsi muscle by increasing the relative abundance of Christensenellaceae_R-7_group in rumen.

Paper mulberry (Broussonetia papyrifera) is a high-quality silage protein feed material that can help address feed shortages and support livestock development. Although some studies have investigated the relationships between microbial communities and silage quality, these relationships and the underlying community assembly processes remain complex, requiring further research to clarify them. Additionally, limited research has explored the relationship between microbial community fermentation functions and silage quality. In this study, we aimed to explore B. papyrifera and wheat bran mixed silage quality driven by the characteristics of the microbial community. After 50 days of silage fermentation, high-quality and low-quality samples were selected from every mixing ratio (90:10, 80:20, and 65:35). The silage chemical composition, lignocellulose degradation enzyme activity, microbial community composition, and potential functions were used to explore the relevance between silage quality and the characteristics of the microbial community. The contents of hemicellulose, neutral detergent fiber, pH, and the activities of endoglucanase and exoglucanase were significantly affected by mixing ratios and silage quality grade. There were higher crude protein content, lignocellulose degrading enzyme activity, and lower pH, lignin, and acid detergent fiber in the mixing of 65:35 (BP65%) samples. The PERMANOVA results showed that mixing ratios had significant impacts on microbial community composition and bacterial fermentation functions. There was a higher bacterial diversity, lower fungal diversity, and better functional potentials for fermentation and lignocellulose degradation in BP65% high-quality silage. The dominant genera were Lactobacillus , Cladosporium , and Wallemia in all samples. The relative abundance of Clostridium , Rhodococcus , Turicibacter , Ralstonia , and Burkholderia was significantly higher in BP65% high-quality samples. There was a higher abundance of Wallemia in the BP65% samples than in other mixing ratios samples. Notably, silage quality showed a close relationship with Lactobacillus , Turicibacter , Romboutsia , Wallemia, and Pichia. In summary, 65:35 was a suitable mixing ratio for B. papyrifera and wheat bran silage, but high-quality silage still required the participation of multiple specific rare microbial taxa. The higher bacterial diversity and specific microbial taxa abundance could be critical for improving B. papyrifera silage quality. We expect that our findings will provide new insights into silage quality driven by the characteristics of the microbial community. [ABSTRACT FROM AUTHOR]

Paper mulberry (Broussonetia papyrifera), as a new woody forage with high-protein characteristic, is being widely used in ruminant feeding. However, little is known about the comprehensive microbiota picture of whole ruminal niches (liquid, solid, and epithelium) under paper mulberry diet. To gain a better understanding of feeding paper mulberry on the rumen microbiota, the effects of fresh paper mulberry, paper mulberry silage, or a conventional high-protein alfalfa silage on rumen fermentation products and microbiota in rumen niches of Hu lambs were studied. Forty-five Hu lambs were randomly divided into 3 treatments with 15 replicates in each treatment. No significant difference was observed among treatments in the average daily gain (ADG). The fresh paper mulberry treatment had lower (P < 0.05) pH and higher (P < 0.05) total volatile fatty acids (TVFA) compared with silage treatments, but the fermentation parameters did not show significant differences between paper mulberry silage and alfalfa silage treatments. The Shannon index did not show a significant difference (P < 0.05) among treatments except between fresh paper mulberry and alfalfa silage treatment in rumen epithelial niches. Butyrivibrio and Treponema were the predominant genera in the rumen epithelial fraction, while Prevotella and Rikenellaceae_RC9 dominated in both rumen liquid and solid fractions. These results indicated the paper mulberry supplement did not have distinct impact on the microbial diversity and growth performance compared with alfalfa silage, especially for paper mulberry silage, which might help us develop an alternative animal feeding strategy of replacing alfalfa with paper mulberry. Key points: • Feeding paper mulberry silage did not show significant impact on the growth performance compared with alfalfa silage treatment. • Feeding fresh paper mulberry reduced rumen pH value and increased total volatile fatty acid. • The microbial diversity did not show significant difference among treatments. [ABSTRACT FROM AUTHOR]

The study aimed to compare the impact of four chemical additives on fermentation characteristics, aerobic stability and in vitro digestibility of total mixed ration (TMR) silage based on soy sauce residue. The TMR (35% soy sauce residue + 45% Napier grass + 20% concentrate) was placed into silos (10 L). The experiment followed the completely random design, treated with different chemical additives: (1) distilled water (control); (2) 0.1% potassium sorbate (SP); (3) 0.1% sodium benzoate (SS); (4) 0.5% calcium propionate (SC); (5) 0.5% sodium diacetate (SD). Total of 100 silos (5 treatments × 4 aerobic exposure days × 5 replicates) were ensiled for 60 days. After exposure to the air, the samples were analyzed for the dynamic change of fermentation parameters at 4, 9 and 15 days, and the data was analyzed as repeated measures. The content of butyric acid and ammonia nitrogen was maintained at a low level. The highest (p < 0.05) lactic acid (LA) content and the lowest (p < 0.05) pH value were measured in SP. At the first 4 days of aerobic exposure, TMR silages treated with four chemical additives were more stable relative to the control, as indicated by the low pH value and yeast counts. Furthermore, the highest (p < 0.05) LA content and the lowest (p < 0.05) pH value indicated that SP performed superior aerobic stability compared with other chemical additives. The SP shows higher (p < 0.05) 72 h cumulative gas production (GP 72 ) and in vitro neutral detergent fiber digestibility (IVNDFD) relative to the control. In conclusion, the SP performed superior in improving fermentation characteristics, aerobic stability and in vitro digestibility of TMR silages based on soy sauce residue., (© 2024 Wiley‐VCH GmbH. Published by John Wiley & Sons Ltd.)

Paper mulberry (Broussonetia papyrifera, PM) is high protein but unutilized as a feed source. The study explores the different parts (leaf, stem, and whole plant) of PM chemical composition, silage fermentation, and in vitro and in situ digestibility, aiming to give some guidelines to PM usage as feed. The result showed that the leaf had a higher fresh weight than the stem (p < 0.05). The dry matter contents of the three groups had no differences. The highest crude protein, ether extract, water-soluble carbohydrate, ash, calcium, phosphorus, amino acid contents, and butter capacity were observed in the leaf (p < 0.05). The stem had the highest (p < 0.05) neutral detergent fiber, acid detergent fiber, and lignin contents. After ensiling, the stem silage had the lowest pH value, ammonia nitrate (NH3-N), lactate, acetate, and propionate (p < 0.05). The leaf silage had the highest pH value (p < 0.05). The lactate, acetate, and propionate in the leaf and whole plant silage had no difference. The butyrate was not detected in all silage. The in vitro and in situ digestibility experiments showed the leaf had the highest digestibility (p < 0.05), which could produce more volatile fatty acids and have a higher effective digestibility. These results allow a greater understanding of PM to be used as a feedstuff.

Objective This study was carried out to investigate the possible application of Broussonetia papyrifera (B. papyrifera) silage as a functional feeding stuff in dairy cattle. Methods Seventy-two Holstein cows were divided into four groups randomly and allocated to 6 pens with 3 individuals in each group and fed the original total mixed ratio (TMR) in the dairy farm or the new TMR with 5%, 10%, and 15% B. papyrifera silage, separately. Feed intake were recorded, milk and blood samples were collected, and milk composition, blood metabolites and milk fatty acids composition were measure at the end of the experiment. Results Dry matter intake of cows decreased when they fed on diet with B. papyrifera, but no differences were observed in body condition score, milk yield, milk protein and lactose, feed efficiency and serum metabolites between groups. Both 10% or 15% of B. papyrifera silage in the diet significantly increased the immunoglobulin A (IgA) and IgG in serum, 15% of B. papyrifera silage increased the content of serum catalase, superoxide dismutase, total antioxidant capacity, and decreased the content of 8-hydroxy-2′-deoxyguanosine. Furthermore, 10% or 15% of B. papyrifera silage resulted in a significant decrease in the milk somatic cell count, and increased the polyunsaturated fatty acids content in the milk. Conclusion The diets with 10% to 15% of B. papyrifera silage might enhance the immune and antioxidant function of dairy cows and increase the polyunstaturated fatty acid concentration in the milk.

Silage quality remains an important issue in farming, as do limitations in the range of products suitable for animal fodder. We therefore explored the microorganisms that are critical for the fermentation quality of paper mulberry silage. Low (unwilted) and high (wilted) dry matter (DM) paper mulberry were harvested at two cutting times. These were ensiled for 0, 3, 7, 14, and 56 days, respectively. Compared with unwilted silages, wilting significantly decreased (p < 0.05) silage pH value, ammonia‐N concentration, and yeast counts but increased (p < 0.05) lactic acid content. In addition, higher (p < 0.05) crude protein (CP) contents were also observed in wilted silages. Next‐generation sequencing (NGS) analysis revealed that wilting reduced the abundance of Enterobacter, while increasing that of Lactobacillus. Single‐molecule real‐time sequencing (SMRT) revealed that the silage was enriched in the lactic acid bacteria (LAB), Lactobacillus rhamnosus after wilting, which showed a positive correlation with CP and lactic acid content. We conclude that wilting may help preserve paper mulberry silage, facilitating its use as a new fodder resource. Moreover, L. rhamnosus has the potential to be developed as a new inoculant for the modulation in wilted silages, particularly paper mulberry silage. [ABSTRACT FROM AUTHOR]

Paper mulberry (Broussonetia papyrifera L.) is an unconventional forage with high crude protein content and is widely used in China. In order to evaluate the pattern of fermentation quality and the aerobic stability when inoculated with lactic acid bacteria, laboratory-scale silage was prepared. The experimental groups included a control group (CK), a Lactobacillus plantarum 'LC365283' (selected from paper mulberry silage) treatment (L1), a commercial inoculant Lactobacillus plantarum treatment (GF), a commercial inoculant Lactobacillus buchneri treatment (FR), a 3% molasses treatment (MO), a 150 U/g cellulase treatment (CE), and their combinations (MO + L1, MO + GF, MO + FR, CE + L1, CE + GF, and CE + FR). The changes in bacterial community and composition of the fermentation products were evaluated after being ensiled for 30 days and unsealed for 1, 3, 5, and 7 days. Compared with the CK, MO and CE, the silages treated with L1, MO + L1, and CE + L1 showed higher lactic acid concentrations, lower pH values, and lower ammonia nitrogen concentrations (p < 0.05). During the first 3 days of aerobic exposure, the pH values and organic acid content changed slightly in all treatments. The present study suggests that addition of L1 was better than commercial inoculum, and the paper mulberry silages could be well preserved after being unsealed for 3 days. [ABSTRACT FROM AUTHOR]

The purpose of this study was to investigate the effects of inoculation with two lactic acid bacteria (LAB) strains (Lacticaseibacillus rhamnosus and Lentilactobacillus buchneri) and the addition of four corn flour proportions (0, 3, 6, and 9%) in different treatments, on the composition and function of the bacterial community in whole-plant paper mulberry silage. The different treatments promoted Lactiplantibacillus, Lentilactobacillus, and Lacticaseibacillus growth, but the microbial species responsible for fermentation differed among the treatments. High species diversity and various Gram-negative bacteria, such as Flavitalea sp., Pantoea agglomerans, Acinetobacter pittii, Turicibacter sanguinis, and Ralstonia pickettii, were found in the uninoculated LAB treatments. A beneficial bacterium, Lactobacillus johnsonii, was discovered for the first time in whole-plant paper mulberry silage. LAB inoculation simplifies the microbial community structure, and beneficial Lactobacillus as a key species aggregates in the inoculated treatment group. However, L. rhamnosus inoculation alone may have limited bacteriostatic activity against in whole-plant paper mulberry silage. Compared with silage lacking corn flour, amino sugar and nucleotide sugar metabolism, galactose metabolism, the phosphotransferase system and the pentose phosphate pathway metabolic activity were increased in corn flourcontaining silage. Whole-plant paper mulberry can be used as a highquality silage to provide high-quality feed resources for sustainable ruminant livestock production. Moreover, additive use is necessary for preparing paper mulberry silage. [ABSTRACT FROM AUTHOR]

In this study, the influence of two lactic acid bacteria (LAB) strains [Lactobacillus rhamnosus BDy (LR-BDy) and Lactobacillus buchneri TSy (LB-TSy)] selected from Southwest China on the fermentation characteristics and in vitro gas production of Broussonetia papyrifera (paper mulberry) silage were experimentally explored. The experimental groups were a control group (C), an LB-TSy treatment (LB), an LR-BDy treatment (LR), and an LR-BDy + LB-TSy hybrid group (LR × LB). After the LAB were added, the pH value of paper mulberry silage significantly declined (p < 0.05), and the crude protein content was effectively preserved (p < 0.05). However, no significant changes were found in the levels of neutral detergent fiber, acid detergent fiber, and crude ash (p > 0.05). The lactic acid content in paper mulberry silage was evidently increased (p < 0.05). The in vitro gas production in the LR at 36, 48, and 72 h were markedly higher than that in the other treatments (p < 0.05). Owing to the addition of LAB, the microbial diversity in paper mulberry silage was reduced, while the relative bacterial abundance of Lactobacillus was enhanced. Hence, the addition of LAB selected from the warm and humid region in Southwest China can improve the quality of paper mulberry silage and elevate its feeding value in this region. [ABSTRACT FROM AUTHOR]

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., (© 2021 Society of Chemical Industry.)

The anaerobic batch digestion of freeze‐dried and ground (< 1.5 mm) maize silage, wheat straw, cattle manure, pig manure, and cellulose filter paper was investigated with substrates placed in polyester filter bags. Gas production dynamics of bagged and non‐bagged substrates were compared. By using batch bottles running in parallel with those used for gas measurement, substrates could be sampled over time and various parameters measured for both substrate and bulk liquid. The bags allowed an immediate mass loss into the bulk liquid with some substrates and a generally lower rate of gas production. This method requires refinement but has potential for the study of the dynamics of substrate degradation during anaerobic digestion. [ABSTRACT FROM AUTHOR]

Globally, climate change is an alarming threat to the livestock industry. Such changes in the climate can also adversely affect the returns of livestock farmers in Pakistan. Improvement in the production process could decrease the risk of losses. This study analyzes the efficacy of silage to abate the losses in livestock profitability resulting from the climate change. The study employed cross-sectional survey data of 492 livestock farmers collected from six districts of Punjab Province, Pakistan. The data are analyzed with endogenous switching regression, considering the possibility of selection bias and endogeneity in adopting silage as a climate change adaptation measure. The study findings show a significant difference in material well-being between silage users and conventional feed users. Furthermore, training programs such as silage training and livestock development programs could be meaningful provisions to abate climate change and improve food security. It is suggested that training and development programs should be incorporated in policy plans to improve the well-being of farmers in terms of their farm revenues., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

This study aimed to investigate the influence of herbal residues on the fermentation quality and ruminal fermentation of paper mulberry silage. Clove, mint, and purple perilla residues were used as additives. Silage treatments were designed as control (no additives), 5% of clove, 5% of mint, and 5% of purple perilla. After 21 and 75 days of fermentation, the fermentation characteristics, bacterial communities, and ruminal greenhouse gas emissions in vitro incubation of paper mulberry were analyzed. The results showed that the used herbal residues could reduce the protein losses in paper mulberry silage based on the lower contents of ammoniacal nitrogen and nonprotein nitrogen. Compared with control, higher lactic acid and propionic acid contents were observed in the silages treated with mint and purple perilla but with a higher acetic acid content in clove treatment. Real-time sequencing technology (single-molecule real-time) revealed that Lactobacillus was the dominant bacteria in all silages at the genus level, whereas the bacterial abundance in the treated silages differed greatly from control at the species level. Lactobacillus hammesii abundance was the highest in control, whereas Lactobacillus acetotolerans was the first predominant in the treated silages. All the additives enhanced the digestibility of in vitro dry matter significantly. However, purple perilla decreased the production of total gas, methane, and carbon dioxide. The findings discussed earlier suggested that herbal residues have potential effects in improving fermentation quality, reducing protein loss, and modulating greenhouse gas emissions in the rumen of paper mulberry silage by shifting bacterial community composition. [ABSTRACT FROM AUTHOR]

Paper mulberry (Broussonetia papyrifera) plants are served as a local roughage in China, and they are mostly processed as silage for ruminants. This study aimed to explore the effects of different silage additives on the chemical composition, fermentation profile, as well as the in vitro and in situ digestibility of paper mulberry (PM) silage. Four groups consisting of PM silage, three with additives and one without any additives as the control group (CON), were established. The three experimental groups with additives were set up as follows: CON with 5 × 106 CFU per gram of fresh PM weight of lactic acid bacteria (Lactobacillus plantarum) (LAB); CON with 3% fresh PM weight of molasses (MOL) added to the PM silage; and CON with both LAB and MOL added (LM). After 45 days of ensiling at 20 °C, all of the PM treatment groups increased their ash content and decreased their water-soluble carbohydrate content (p < 0.05). Meanwhile, the pH and NH3-N content of the PM silage were lower in the additive treatment groups than in the CON group (p < 0.05). Lactic acid in the LM group was the highest (p < 0.05) among the four groups, and trace amounts of butyric acid was detected only in the CON group. In vitro dry matter digestibility was similar among all groups. Results of the in situ experiment found that the effective digestibility of the PM silage dry matter, as well as the acid detergent fiber digestibility was higher in the LM group than in the CON group (p < 0.05). In conclusion, the addition of LAB, MOL, and their combination can improve PM silage fermentation and improve the in situ digestibility of dry matter and acid detergent fiber; however they do not affect in the vitro digestibility of PM silage. [ABSTRACT FROM AUTHOR]

There are few studies on the application of lactic acid bacteria in the reduction of anti-nutrient factors in paper mulberry silage. This study aimed to investigate the effects of different lactic acid bacteria on the fermentation quality and the amount of anti-nutritional factors in paper mulberry silage. Two strains of Lactobacillus plantarum (GX, isolated from paper mulberry silage; GZ, provided by Sichuan Gaofuji Biotechnology Co. Ltd.) were added as silage additives. On days 7, 15, 30 and 60 of the ensiling process, the fermentation quality, and the amount of anti-nutritional factors were measured. Compared with the control group, inoculation with Lactobacillus plantarum could rapidly reduce pH values, leading to lower NH3-N/TN. Besides, it also significantly increased the lactic acid content (p < 0.05). The two strains of L. plantarum significantly reduced the content of hydrolysed tannin, condensed tannin, total tannin, oxalic acid, phytic acid and saponin (p < 0.05). Overall, this study found that the addition of lactic acid bacteria could significantly improve the fermentation quality of paper mulberry and reduce the amount of anti-nutrient factors (p < 0.05). [ABSTRACT FROM AUTHOR]

Lactic acid bacteria occupy an important position in silage microorganisms, and the effects of exogenous lactic acid bacteria on silage quality have been widely studied. Microbial metabolism has been proved as an indicator of substrate utilization by microorganisms. Paper mulberry is rich in free carbohydrate, amino acids, and other components, with the potential to be decomposed and utilized. In this study, changes in the microbial metabolism characteristics of paper mulberry silage with Lactiplantibacillus plantarum (LP) and Lentilactobacillus buchneri (LB) were studied along with a control (CK) using BIOLOG ECO microplates. The results showed that average well-color development (AWCD), Shannon diversity, Shannon evenness, and Simpson diversity exhibited significant temporal trends. LB and LP responded differently in the early ensiling phase, and the AWCD of LB was higher than LP at 7 days. Principal component analysis revealed that CK, LB, and LP samples initially clustered at 3 days and then moved into another similar cluster after 15 days. Overall, the microplates methodology applied in this study offers important advantages, not least in terms of accuracy. [ABSTRACT FROM AUTHOR]

Background: Paper mulberry has been considered as a high-quality protein feedstuff to cope with the shortage of feed and the development of livestock. In addition, the features of high moisture and low water-soluble carbohydrate concentration in fresh paper mulberry make it difficult to ensile. Therefore, it is important to find an optimal way to improve the paper mulberry silage quality. In this study, we aimed to investigate the application of Lactobacillus plantarum (LP) and wheat bran (WB) on the fermentation characteristics, chemical composition and microbial community of paper mulberry silage. Results: The objective of this study was to evaluate the effects of Lactobacillus plantarum and wheat bran alone or combination (LP + WB) addition on the fermentation quality and bacterial community of paper mulberry silage. After 60 days of ensiling, the employed three treatments had higher crude protein contents compared with control (P < 0.05). More importantly, WB and LP + WB treatments significantly reduced the pH value and NH3-N concentration, and increased lactic acid content (P < 0.05). Microbial analysis indicated that the bacterial community in WB and LP + WB treatments showed distinct difference with LP and control. Lactobacillus was the dominant genera in all treatments. However, at the species level, Lactobacillus farciminis became the most dominant bacteria in control and LP treatments while the dominant bacteria in WB and LP + WB were Lactobacillus brevis and Lactobacillus farciminis. In addition, Lactobacillus brevis was positively correlated to crude protein and lactic acid and negatively correlated to pH and NH3-N. Overall, this study revealed that ensiling paper mulberry with WB or combination LP could improve silage quality through altering microbial community, which provided a practical approach for enhancing paper mulberry silage quality. Conclusion: Wheat bran and combinations of Lactobacillus plantarum and wheat bran additions could reduce pH, NH3-N and increase LA content. The application of WB and LP + WB shifted the dominant bacteria species to Lactobacillus brevis. In summary, the addition of wheat bran and combinations of lactic acid bacteria and wheat bran were effective ways to enhance paper mulberry silage fermentation. [ABSTRACT FROM AUTHOR]

With increasing demand for high-quality pork, development of green and healthy feed for finishing pigs is urgently needed. In this study, the effects and mechanisms of mulberry and paper mulberry silages on growth performance, meat quality, and intestinal health of finishing pigs were explored. Intestinal microbiota were profiled, and microbially produced short-chain fatty acids (SCFAs) were measured. The average daily gain (ADG) and feed conversion rate (FCR) with mulberry and paper mulberry silages were not significantly different from those of the control. Meat quality as measured by pork marbling and fatty acids in the longissimus dorsi was better with mulberry silage. The highest concentration of SCFAs was also with mulberry silage. According to 16S rRNA sequencing, Clostridium&#95;sensu&#95;stricto&#95;1 , Terrisporobacter , and Lachnospiraceae , which are important in SCFA production, were biomarkers of mulberry silage. PICRUSt functional analysis of intestinal microbes indicated that galactose metabolism, starch and sucrose metabolism, and carbohydrate digestion and absorption decreased significantly in silage treatments but increased in the control. Correlations between intestinal microbes and SCFAs and fatty acids indicated Clostridium&#95;sensu&#95;stricto&#95;1 , Terrisporobacter , and Lachnospiraceae were closely associated with SCFA and fatty acid contents. The results indicated that mulberry silage could increase SCFA content through shaping intestinal microbes to affect the deposition of fatty acids, which laid a solid theoretical foundation for improving pork quality. IMPORTANCE To avoid competition between people and animals for food, it is essential to develop nontraditional feeds. In this study, the effects of the silages of the unconventional feed resources mulberry and paper mulberry on meat quality of finishing pigs were examined. With mulberry silage in the diet, meat quality improved as indicated by meat color, marbling score, and beneficial fatty acids in the longissimus dorsi muscle. Pigs fed mulberry silage had the highest concentrations of short-chain fatty acids (SCFAs), and 16S rRNA sequencing identified Clostridium&#95;sensu&#95;stricto&#95;1 , Terrisporobacter , and Lachnospiraceae as biomarkers, which are important in SCFA production. Functions of intestinal microbes in the two silage groups primarily involved amino acid metabolism and SCFA production. Correlations between intestinal microbes and SCFAs and fatty acids indicated that Clostridium&#95;sensu&#95;stricto-1 , Terrisporobacter , and Lachnospiraceae were closely associated with SCFA contents in the intestine and fatty acids in the longissimus dorsi.

Dairy cows often suffer metabolic disorders due to the challenges of the transition period. The aim of study was to determine the effects of feeding pomegranate by-product silage (PBS) on feed intake and some blood parameters, such as non-esterified fatty acids (NEFA) and beta hydroxy butyric acid (BHBA), interleukin-1 (IL-1), serum amyloid-A (SAA), prostaglandin F 2α metabolites (PGFM), and progesterone (P4) in fresh Holstein cows. The experimental groups were as follows: CON (0 g PBS of DM) and PBS (120 g PBS of DM). Results showed that replacing corn silage with PBS had no effect on dry matter intake (DMI) throughout the study. The CON group had highest (p < .05) NEFA at 14 d postpartum and serum BHBA was decreased by feeding PBS, especially on days 21 (p < .05) and 28 (p < .05) and the entire study (p < .01). Cows fed PBS had lower (p < .05) serum Malondialdehyde (MDA) than CON cows at different times. Feeding PBS decreased (p < .01) serum IL-1 compared to CON group (216.7 versus. 515.5 Pg/ml) at 28 d postpartum, and also cows fed 120 g PBS had higher (p < .01) serum P4. It was concluded that feeding PBS had no adverse effect on feed intake, milk production, and some reproductive parameters. Moreover, PBS positively affected serum metabolites in fresh dairy cows., (© 2020 Japanese Society of Animal Science.)

Developing and utilizing more roughage resources is conducive in improving the health status of dairy cows and beneficial in controlling the production costs of raw milk. In the current study, we investigated the impact of replacing alfalfa hay with paper mulberry silage (PMS) on the production performance of low lactating dairy cows. Sixty multiparous Holstein cows were stratified by milk yield, days in milk and parity and randomly assigned to 4 treatments: control diet without PMS (CON) or 33.33 %, 66.67 % and 100 % alfalfa hay replaced by an equal amount of PMS (DM basis, PMS1, PMS2, and PMS3, respectively). The experiment lasted for 14 weeks, with the first 2 weeks for adaptation. Dry matter intake of low lactating dairy cows was not affected by PMS, and though milk yield of PMS3 was 1 kg lower than CON, no statistical differences were observed. Lactose yield and feed conversion efficiency (FCE) tended to be affected by treatment × week interaction (P = 0.07 and P = 0.08, respectively), and the FCE of PMS groups was lower than CON in wk12 (P = 0.08). No treatment effect on oxidative stress-related variables was detected, except malondialdehyde (MDA), which tended to be decreased by PMS (P = 0.06). Cows in PMS3 had lower CP digestibility than CON (P = 0.004), and the apparent digestibility of NDF and ADF were lower for PMS2 compared to CON (P = 0.004 and P = 0.05, respectively) and PMS1 (P = 0.005 and P = 0.006, respectively). In conclusion, replacing alfalfa hay with paper mulberry in the diet of low lactating dairy cows did not affect DMI and milk yield, but improved their antioxidative capacity. However, the amount of paper mulberry should be added to the diet cautiously, due to its potentially adverse effect on digestibility and milk yield in dairy cows. • Dry matter intake, milk yield and milk components were not affected by paper mulberry silage. • Paper mulberry silage can improve the antioxidative capacity of dairy cows. • The apparent digestibility of CP, NDF and ADF was decreased by paper mulberry silage • As a new high-protein forage source, paper mulberry silage can replace alfalfa hay used in low lactating cow diet. [ABSTRACT FROM AUTHOR]

In this study, we aimed to comprehensively evaluate the effects of prepartum replacement of corn silage and distillers' grain diets with hay on the postpartum health and production performance in dairy heifers. Pregnant Holstein dairy heifers were randomly assigned to two groups and fed different diets with the same net energy and protein concentrations. The low corn silage plus hay (CH) group diet mainly contained oat hay (28.5%), corn silage (25%) and folium ginkgo (23%). The high corn silage plus distillers' grains (CD) group diet contained corn silage (53.5%), distillers' grains (5%) and folium ginkgo (18%). Dairy heifers were fed from the beginning of pregnancy to the end of the transition period. Calf birthweight, postpartum morbidity, milk production and reproductive performance were determined. Compared with the cows in the CH group, those in the CD group presented increased calf birthweight and a high risk of some postpartum disorders (p < 0.05), particularly ketosis (37.63% vs. 24.54%) and retained placenta (9.41% vs. 3.92%). However, there was no difference between the groups in reproductive performance, including first timed artificial insemination (TAI) rate, date of TAI and conception rate of TAI. The lactation curves showed no difference in milk yield during the peak lactation period of primiparous cows between the CH and CD groups; the total milk production was 33.39 kg and 33.19 kg respectively. Notably, the time to peak milk production was delayed in the CD group. It can be concluded that replacing corn silage and distillers' grains with hay can reduce postpartum morbidity in dairy heifers and shorten the time to peak milk production., (© 2022 Wiley-VCH GmbH.)

Aims: To investigate the fermentation characteristics, bacterial community and predicted functional profiles during the ensiling of wilted alfalfa (Medicago sativa L.)., Methods and Results: First-cutting alfalfa was harvested at the early bloom stage, wilted for 6 h, and ensiled in laboratory-scale silos (1 L capacity). Triplicate silos were sampled after 1, 3, 7, 15, 30 and 60 days of ensiling, respectively. The bacterial communities of wilted alfalfa and silages on day 3 and 60 were assessed through high throughput sequencing technology, and their functional characteristics were evaluated according to the Kyoto Encyclopedia of Genes and Genomes using Tax4Fun. After 60 days of ensiling, alfalfa silage showed a moderate fermentation quality, indicated by high lactic acid (56.7 g kg -1 dry matter [DM]) and acetic acid (39.4 g kg -1 DM) contents, and low concentrations of butyric acid (2.12 g kg -1 DM) and ammonia nitrogen (128 g kg -1 total nitrogen). Lactobacillus rapidly became predominant on day 3 and increased to 60.4% on day 60. Results of functional prediction analyses showed that the metabolism of amino acid, energy, cofactors and vitamins were reduced, while metabolism of nucleotide and carbohydrate were increased during ensiling. Fructokinase, 1-phosphofructokinase and pyruvate kinase played important roles in producing lactic acid. The production of acetic acid may be correlated with the enhancement of 6-phosphogluconate dehydrogenase and acetyl-CoA synthetase., Conclusions: Knowledge regarding bacterial dynamics and their metabolic pathways during alfalfa ensiling is important for understanding the fermentation process and may contribute to the production of nutritious and stable alfalfa silage., Significance and Impact of the Study: High throughput sequencing technology combined with 16S rRNA gene-predicted functional analyses could provide a new and comprehensive insight into bacterial community dynamics and functional profiles to further improve the silage quality., (© 2021 The Society for Applied Microbiology.)

This study was aimed to investigate the effect of storage time on fermentation characteristics, bacterial community structure and predicted metabolic pathways of napiergrass (Pennisetum purpureum Schum.) silage. First-cutting napiergrass was harvested at the vegetative stage and ensiled in laboratory-scale silos (1 L capacity). Triplicate silos were sampled after 1, 3, 7, 15, 30 and 60 days of ensiling, respectively. The bacterial communities on day 3 and 60 were assessed through the high throughput sequencing technology, and metabolic pathways of bacterial community were predicted according to the Kyoto Encyclopedia of Genes and Genomes (KEGG) via Tax4Fun. Napiergrass silage exhibited an acetic acid-type fermentation, indicated by lower lactic acid contents and ratio of lactic acid to acetic acid, and higher pH, ethanol and acetic acid contents. Before ensiling, the predominant genera in fresh napiergrass mainly included Acinetobacter, Enterobacteriaceae, Enterobacter and Lactococcus. After 60 days of ensiling, high proportions of Enterobacteriaceae, Enterobacter and Lactobacillus were found in napiergrass silages. The metabolism of amino acid, energy, cofactors and vitamins were inhibited, whereas metabolism of nucleotide and carbohydrate were promoted during ensiling. Overall, the combination of high throughput sequencing technology and 16S rRNA gene-predicted functional analyses revealed the differences during the initial and late stages of napiergrass silages not only for distinct bacterial community but also for specific functional metabolites. It could provide a comprehensive insight into bacterial community and functional profiles to further improve the silage quality., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Aims: To effectively use woody plant resources to prepare silage for ruminants, Pacific Biosciences (PacBio) single-molecule real-time (SMRT) sequencing was applied to study the microbial network and fermentation characteristics of paper mulberry (PM) silage prepared with corn meal (CM) and rice bran (RB) as exogenous additives., Methods and Results: PM is rich in nutrients and contains more than 26% crude protein in dry matter. After ensiling, the microbial diversity and abundance in PM, CM and RB decreased due to the anaerobic environment and acidic conditions. The CM-treated PM silage accelerated the conversion of the dominant microbial community from harmful bacteria to lactic acid bacteria and promoted lactic acid fermentation. When RB was used to treat PM silage, Enterobacter and Clostridium species became the main bacterial community during ensiling, leading to butyric acid fermentation and protein decomposition. Compared with RB, CM increased the amount of fermentation substrates, changed the microbial community structure and affected metabolic pathways (global metabolism, carbohydrate metabolism and amino acid metabolism), which improved the flavour and quality of the PM silage., Conclusions: The CM addition of improved the fermentation quality of PM silage, with PM + CM being the ideal combination. The SMRT sequencing technology could accurately obtain specific details of the microbial networks and fermentation characteristics. Our results indicate that PM can be used as a potential high-protein silage in animal production., Significance and Impact of the Study: In tropics, the effective use of abundant natural biomass resources such as woody plants to prepare silage for feed preservation can solve the problem of restricting livestock production due to the shortage of feed in the dry season. SMRT sequencing technology was used to accurately analyze the microbial network and fermentation characteristics of woody silage prepared with CM as an exogenous additive to improve the fermentation quality of silage., (© 2021 The Society for Applied Microbiology.)

This study compared ensiled sorghum (ES) from two varieties (first and second cuts) with corn silage (CS) for chemical analysis, total-tract nutrient digestibility, ruminal variables and blood biochemistry metabolites in sheep. Five experimental silages representing first cut ES var. Pegah (PS1), second cut ES var. Pegah (PS2), first cut ES var. Speedfeed (SS1), second cut ES var. Speedfeed (SS2) and CS were assessed for chemical analysis, silage fermentation and anti-quality compounds. Diets consisted of lucerne hay, silage and a premix (at the ratio of 53.5:46:0.5 on a dry matter [DM] basis). Feed consumption, total-tract nutrient digestibility, microbial nitrogen supply (MNS), ruminal variables and blood biochemistry metabolites were determined in a Latin square design of five periods with five male sheep. Compared to sorghum, CS had lower (p < 0.01) concentrations of ash, water-soluble carbohydrates and crude protein, but higher starch, lignin (sa), non-structural carbohydrates and apparent nutrient digestibility. The measurements of pH, lactic acid, NH 3 -N and volatile fatty acids (VFA) confirmed that all silages were well-preserved. The anti-quality compound levels in the silages were below the safe limits for ruminants. No differences were observed in feed consumption (p > 0.05) and total-tract digestibility of DM and nutrients (p > 0.05). Rumen pH, NH 3 -N, VFA concentrations and acetate to propionate ratio showed no difference among treatments (p > 0.05). Sheep fed on sorghum silage showed a decrease (p < 0.01) in total protozoa, and particularly in the Entodiniae population. There were no differences in cellulolytic bacteria counts (p > 0.05) among treatments. Urinary N, MNS and blood biochemistry metabolites showed no difference among treatments (p > 0.05). Silages from both cuts of sorghum had comparable total-tract nutrient digestibility to CS, therefore, these varieties could be used as a more drought resilient silage source in the feeding of sheep., (© 2021 Wiley-VCH GmbH.)

In order to effectively use locally available woody forage and hay resources in tropics, paper mulberry (PM) silages were prepared using Napier grass (NG) and rice straw (RS) hays. The mixing proportion of hays were 10%, 20%, and 30% on fresh matter basis. After 60 days of ensiling, single molecule, real-time (SMRT) sequencing technology was used to analyse the dynamic changes of microbial community and silage fermentation. When PM silages were prepared with both hays, the moisture of PM were adjusted below 67.6%. The dominant microbial community rapidly shifted from Gram-negative to Gram-positive bacteria during ensiling. Before ensiling, aerobic bacteria were dominant in materials, but after ensiling, Lactobacillus plantarum became the main bacterial community and dominated the fermentation process. The result of Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis shown that hay-treated PM silages increased the proportion of Global and overview maps and carbohydrate metabolic pathways, while decreasing the proportion of amino acid metabolic pathway. Fatty acid metabolism, glycolysis and proteolysis categories may improve the flavour and quality of the silage. PM silage prepared with 10% hay effectively improved better fermentation quality than PM alone silage. The result confirmed that PM and hay are the ideal combination, they can prepare good-quality silage for ruminant feed to alleviate the shortage of feed in tropics. • Napier grass and rice straw improve fermentation suitability of paper mulberry. • Paper mulberry with 10% hay are the ideal combination for silage preparation. • Microbial metabolic pathways improve the flavour and quality of the silage. • Hay addition increases the proportion of carbohydrate metabolic pathway in silage. • SMRT technology accurately revealed specific microbial information of silage. [ABSTRACT FROM AUTHOR]