Numerous studies have demonstrated that Broussonetia papyrifera is an unconventional feed resource with significant developmental potential. This research aimed to explore the effects of Broussonetia papyrifera silage on the growth performance, blood parameters, immunity, antioxidation, cytokine levels, and rumen bacterial composition of Kazakh lamb. Forty healthy male Kazakh lambs, aged 5 months and weighing 30.12 ± 1.14 kg, were randomly divided into control and experimental groups, each consisting of four replicates (five lambs per replicate). The control group was fed a basal diet, while the experimental group received a diet supplemented with 20% Broussonetia papyrifera silage (dry matter basis). Following a 10-day pre-feeding period, a 60-day formal experiment was conducted. The results indicated no significant difference in growth performance between the experimental and control groups. However, compared to the control group, the use of Broussonetia papyrifera silage significantly reduced (p < 0.05) neutrophil, lymphocyte, and eosinophil counts, as well as creatinine levels in the blood. Furthermore, Broussonetia papyrifera silage (p < 0.01) enhanced total serum antioxidant capacity, superoxide dismutase, catalase, glutathione peroxidase, immunoglobulin A, immunoglobulin M, immunoglobulin G, interleukin-2, interleukin-6, and interleukin-8, and decreased malondialdehyde and interleukin-4 levels. Additionally, the use of Broussonetia papyrifera silage increased the diversity and richness of the rumen bacterial community, notably enhancing the relative abundance of Firmicutes such as Rikenellaceae_RC9_gut_group and Christensenellaceae_R-7_group. In conclusion, feeding Kazakh lamb with Broussonetia papyrifera silage (20% DM) did not adversely affect their growth performance but improved their immunity and antioxidant capacity and enhanced the relative abundance of beneficial bacteria in the rumen, thereby promoting animal health.

The utilization of nettle (Urtica cannabina) as feed is restricted by its material properties (antibacterial activity and high buffering capacity). This study hypothesized that the use of lactic acid bacteria (LAB) attached to nettles can improve these problems. Lactococcus garvieae (LG), Pediococcus pentosaceus (PP), and LG + PP (LP) isolated from nettles were inoculated into nettle silage to explore nutrient retention and the microbial community structure. The results showed that inoculation significantly delayed dry matter and crude protein loss, inhibited neutral detergent fiber and acid detergent fiber degradation, and reduced ammonia nitrogen (NH₃-N) accumulation. There was a significant increase in Firmicutes abundance after inoculation, and the dominant genus, Aerococcus, was negatively correlated with NH₃-N accumulation. In the later stages of the PP treatment, Atopistipes synergistically inhibited Clostridia with acetic acid. However, the high buffering capacity and antibacterial components of raw nettle led to increased pH values during the later fermentation stages, limiting sustained acid production by LAB. These results confirm that nettle-derived LAB can effectively improve the quality of silage by regulating the microbial community and the acidification process; however, they must be combined with pretreatment strategies or optimized composite microbial agents to overcome raw material limitations. This study provides a theoretical basis and technical support for the utilization of nettle as feed.

Lactic acid bacteria (LAB) and cellulase have been used as additives to improve the fermentation quality of mulberry silage. This study investigated the dynamics of fermentation characteristics and bacterial communities during 60-day ensiling through three established treatment groups: Control (no inoculation), Lactiplantibacillus plantarum (LP) inoculation as well as combination of L. plantarum and cellulase inoculation group (LPC). The results showed that compared with the Control group, the LP and LPC treatments significantly reduced the loss of dry matter, soluble carbohydrates, and crude protein (p < 0.05), effectively promoted the accumulation of lactic acid and acetic acid (p < 0.05), but significantly elevated ammonia nitrogen (NH₃-N) production. Inoculation was beneficial to the stability of the bacterial community in mulberry branch and leaf silage because it can maintain a high level of beneficial bacteria (Lactiplantibacillus) and inhibit the growth of harmful bacteria (Escherichia-Shigella). The combination of the inoculation of L. plantarum and cellulase may improve the quality of mulberry branch silage.

Paper mulberry (Broussonetia papyrifera) is a high-protein forage, but its high buffering capacity and low water-soluble carbohydrate (WSC) content hinder silage fermentation. This study evaluated the effect of tea tree (Melaleuca alternifolia) essential oil (TTO) as a natural additive for paper mulberry silage. TTO was added at 0 (CK), 500 (CSD), and 1000 mg/kg (CSG) fresh weight, with samples collected on days 7, 15, 30, and 90. After 90 days, CSD had the lowest pH and the highest lactic acid content. NH₃-N/TN was significantly lower than in CK (p < 0.05), and the retention of dry matter, crude protein and water-soluble carbohydrate was improved (p < 0.05). Compared with CK, CSD exhibited the highest lactic acid bacteria (LAB) counts, along with the lowest mold and aerobic bacterial counts (p < 0.05). However, CSG did not further improve fermentation quality, and most of the indicators were not significantly different from CSD (p > 0.05). The in vitro gas production test showed that CSD significantly increased the theoretical maximum gas production and the maximum gas production rate (p < 0.05). Additionally, 16S rRNA sequencing revealed a distinct bacterial succession in CSD, which enriched beneficial lactic acid bacteria (LAB) such as Pediococcus while outcompeting less competitive taxa. This microbial reshaping redirected the community toward acidification and nutrient retention. Collectively, these results demonstrate that TTO at 500 mg/kg is a promising natural additive for improving paper mulberry silage quality through selective reshaping of the bacterial community.

Background: Nettle is a potential non-conventional feed resource due to its high level of crude protein content, and ensiling is better for utilization in the animal industry. Previous integrated analysis (microbiome and metabolome) suggested that 4-pentenoic acid and malic acid in ensiled nettle may inhibit harmful microorganisms within the system. The present study investigated the effects of these two acids on nettle silage quality through the addition of 1% fresh weight of these acids, then analyzed the characteristics and bacterial communities during 60 days of nettle ensiling. Results: The addition of 4-pentenoic acid increased the content of crude protein (CP) and acetic acid (AA) compared with both control and malic acid-treated groups during 30 to 60 days of ensiling (p < 0.05). Lactic acid (LA) content was highest in the malic acid-treated group (4.21%, dry matter, DM based) compared to the control and 4-pentenoic acid-treated groups after 7 days of nettle ensiling (p < 0.05), but lower compared with the 4-pentenoic acid-treated group after 30 days of nettle ensiling (p < 0.05), and it was not detected in all groups after 60 days of silage. The contents of butyric acid (BA) and ammonia (AN) were the lowest (2.92–4.39% of DM and 9.94–24.28% of total nitrogen, respectively) in the 4-pentenoic-treated group compared with both control and malic acid-treated groups during 30 to 60 days of ensiling (p < 0.05). Both acids increased the relative abundance of Weissella after 30 days of nettle ensiling, with 4-pentenoic acid showing a higher inhibitory capacity. Both acids showed a trend to inhibit the relative abundance of Clostridium sensu stricto 15 after 30 days of nettle ensiling. Clostridium sensu stricto 15 showed significant positive correlation with BA and AN (p < 0.05). Conclusions: The results of the present study suggested that the addition of 4-pentenoic could improve the quality of silage by reducing levels of protein degradation, probably resulting from its inhibited activity against Clostridium spp. However, malic acid was less effective than 4-pentenoic acid in suppressing Clostridium spp. activity and the associated production of BA and AN, resulting in inferior preservation of CP.

[Background] Nettle is a potential non-conventional feed resource due to its high level of crude protein content, and ensiling is better for utilization in the animal industry. Previous integrated analysis (microbiome and metabolome) suggested that 4-pentenoic acid and malic acid in ensiled nettle may inhibit harmful microorganisms within the system. The present study investigated the effects of these two acids on nettle silage quality through the addition of 1% fresh weight of these acids then analyzed the characteristics and bacterial community during 60 days of nettle ensiling. [Results] Addition with 4-pentenoic acid increased the content of crude protein (CP) and acetic acid (AA) when compared with both control and malic acid treated during 30 to 60 days of ensiling (p＜0.05). Lactic acid (LA) content was highest in malic acid treated (4.21 %, dry matter, DM based) when compared with control and 4-pentenoic acid treated after 7 days of nettle ensiling (p＜0.05), but lower when compared with 4-pentenoic acid treated after 30 days of nettle ensiling (p＜0.05), and was not detected in all groups after 60 days of silage. The content of butyric acid (BA) and ammonia (AN) was lowest (2.92%—4.39% of DM and 9.94 % —24.28% of total nitrogen, respectively) in 4-pentenoic treated when compared with both control and malic acid treated during 30 to 60 days of ensiling (p＜0.05). Both acids treated increased relative abundance of Weissella after 30 days of nettle ensiling, especially for 4-pentenoic acid showed the higher capacity. Both acids treated showed trend to inhibited relative abundance of Clostridium sensu stricto 15 after 30 days of nettle ensiling. Clostridium sensu stricto 15 showed significant positive correlation with BA and AN (p＜0.05). [Conclusion] Present study results suggested that addition of 4-pentenoic could improve quality of silage due to reduction level of protein degradation probably resulting from its inhibited activity against to Clostridium spp. However, malic acid was less effective than 4-pentenoic acid in suppressing Clostridium spp. activity and the associated production of BA and AN, resulting in inferior preservation of CP.