Project description:Inoculation with homofermentative lactic acid bacteria (LAB) effectively enhances the silage quality of forages. Moreover, feeding such LAB-inoculated silage modulates rumen microbiota composition and metabolites, thereby improving ruminant production performance. Nevertheless, the specific mechanism through which LAB inoculants regulate the silage–rumen–mammary gland axis remains unclear.Inoculation with homofermentative Lactiplantibacillus plantarum BX62 improved the alfalfa silage quality. Dairy goats fed the BX62 group silage showed significantly higher milk fat content compared to the control group (no inoculation) (P < 0.05). Integrated analysis of silage microbial metabolomics and experimental validation revealed a significant increase in flavonoid content in the BX62 silage. This was attributed to microbial community restructuring and secretion of carbohydrate-active enzymes (CAZymes), which facilitated plant cell wall degradation and flavonoid release. Rumen metagenomic assembly and binning indicated that feeding flavonoid-rich BX62 silage induced the proliferation of flavonoid-degrading microbes and reshaped the rumen microbiota, which resulted in the upregulation of CAZymes and energy metabolic pathways (e.g., ko00620 Pyruvate metabolism), and enhanced fiber degradation and volatile fatty acid (VFA) production in the rumen. Consequently, acetate-dependent milk fat synthesis was promoted in BX62 group goats as showed by the elevated expressions of acetyl-CoA carboxylase 1 (ACC1), fatty acid synthase (FASN) and acyl-CoA synthetase short-chain family member 2 (ACSS2) in mammary gland. Moreover, four milk fat-positively correlated bacteria species (Eggerthellaceae bacterium, Clostridioides difficile, Candidatus Limivicinus sp., and Collinsella aerofaciens) harboring flavonoid-degrading genes proliferated with elevated flavonoid concentrations in the rumen. In vitro trial further confirmed flavonoid degradation capability in both C. difficile and A. equolifaciens (family Eggerthellaceae), and dose-dependent growth promotion in A. equolifaciens. These results demonstrate that silage-derived flavonoids drive rumen microbiome remodeling and promote mammary lipogenesis through a silage-rumen microbiota-mammary gland triad mechanism.

Project description:Inoculation with homofermentative lactic acid bacteria (LAB) effectively enhances the silage quality of forages. Moreover, feeding such LAB-inoculated silage modulates rumen microbiota composition and metabolites, thereby improving ruminant production performance. Nevertheless, the specific mechanism through which LAB inoculants regulate the silage–rumen–mammary gland axis remains unclear.Inoculation with homofermentative Lactiplantibacillus plantarum BX62 improved the alfalfa silage quality. Dairy goats fed the BX62 group silage showed significantly higher milk fat content compared to the control group (no inoculation) (P < 0.05). Integrated analysis of silage microbial metabolomics and experimental validation revealed a significant increase in flavonoid content in the BX62 silage. This was attributed to microbial community restructuring and secretion of carbohydrate-active enzymes (CAZymes), which facilitated plant cell wall degradation and flavonoid release. Rumen metagenomic assembly and binning indicated that feeding flavonoid-rich BX62 silage induced the proliferation of flavonoid-degrading microbes and reshaped the rumen microbiota, which resulted in the upregulation of CAZymes and energy metabolic pathways (e.g., ko00620 Pyruvate metabolism), and enhanced fiber degradation and volatile fatty acid (VFA) production in the rumen. Consequently, acetate-dependent milk fat synthesis was promoted in BX62 group goats as showed by the elevated expressions of acetyl-CoA carboxylase 1 (ACC1), fatty acid synthase (FASN) and acyl-CoA synthetase short-chain family member 2 (ACSS2) in mammary gland. Moreover, four milk fat-positively correlated bacteria species (Eggerthellaceae bacterium, Clostridioides difficile, Candidatus Limivicinus sp., and Collinsella aerofaciens) harboring flavonoid-degrading genes proliferated with elevated flavonoid concentrations in the rumen. In vitro trial further confirmed flavonoid degradation capability in both C. difficile and A. equolifaciens (family Eggerthellaceae), and dose-dependent growth promotion in A. equolifaciens. These results demonstrate that silage-derived flavonoids drive rumen microbiome remodeling and promote mammary lipogenesis through a silage-rumen microbiota-mammary gland triad mechanism.

Project description:Two-stage two-phase biogas reactor systems consisting each of one batch downflow hydrolysis reactor (HR, vol. 10 L), one process fluid storage tank (vol. 10 L), and one downstream upflow anaerobic filter reactor (AF, vol. 10 L), were operated at mesophilic (M, 37 °C) and thermophilic (T, 55 °C) temperatures and over a period of > 750 d (Figure 1, Additional file 1). For each reactor system and for each process temperature, two replicates were conducted in parallel, denominated further as biological replicates. Further process details were as previously published. Start-up of all fermenters were performed using liquid fermenter material from a biogas plant converting cattle manure in co-digestion with grass and maize silage and other biomass at varying concentrations and at mesophilic temperatures. Silage of perennial ryegrass (Lolium perenne L.) was digested as sole substrate in batches of varying amounts with retention times of 28 d (storage of bale silage at -20 °C, cutting length 3 cm, volatile substances (VS) 32 % of fresh mass (FM), total Kjeldahl nitrogen 7.6 g kgFM-1, NH4+-N 0.7 g kgFM-1, acetic acid 2.6 g kgFM-1, propionic acid < 0.04 g kgFM-1, lactic acid 2.6 g kgFM-1, ethanol 2.2 g kgFM-1, C/N ratio 19.3, chemical oxygen demand (COD) 357.7 g kgFM-1, analysis of chemical properties according to [6]. No spoilage was observed in the silage. Biogas yields were calculated as liters normalized to 0 °C and 1013 hPa (LN) per kilogram volatile substances (kgVS). For chemical analysis, samples were taken from the effluents of HR and AF. For sequencing of 16S rRNA gene amplicon libraries, microbial metagenomes, and microbial metatranscriptomes, samples were taken from the silage digestate in the HR digested for 2 d. At this time point, high AD rates were detected as indicated by the fast increase of volatile fatty acids (VFA), e.g., acetic acid. Sampling was performed at two different organic loading rates (OLR), i.e., batch-fermentation of 500 g (denominated as “low OLR”, samples MOLR500 and TOLR500) and 1,500 g silage (denominated as “increased OLR”, samples MOLR1500 and TOLR1500).

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