Effects of Intrinsic Tannins on Metabolome During Sainfoin Ensiling

Condensed tannins (CT) from sainfoin have a high capacity to inhibit proteolysis. The objective of the present study was to investigated the effects of CT (following supplementation of deactivated CT with polyethylene glycol [PEG]) on metabolome during sainfoin ensiling. As results, in total, 510 metabolites were identified after 60 d of sainfoin ensiling, with 33 metabolites were annotated in the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Among those metabolites, phospholipids were the most abundant (72.7% of total 33 metabolites). In addition, 10 up-regulated and 23 down-regulated metabolites, respectively, were identified in the PEG treated group when compared with the control group, after 60 d of ensiling (p \u3c 0.05). Pediococcus (correlated with 20 metabolites, R2 \u3e 0.88, p \u3c 0.05) and Lactobacillus (correlated with 16 metabolites, R2 \u3e 0.88, p \u3c 0.05) were the bacteria most correlated with metabolites. The results suggested antagonistic effects between Lactobacillus and Pediococcus during ensiling. The proteolysis decreased partly due to CT inhibiting Pediococcus activity during ensiling, with Pediococcus being significantly and positively correlated with dopamine after 60 d of ensiling (R2=0.8857, p \u3c 0.05)

Effects of Intrinsic Tannins on Metabolome During Sainfoin Ensiling

Condensed tannins (CT) from sainfoin have a high capacity to inhibit proteolysis. The objective of the present study was to investigate the effects of CT (following supplementation of deactivated CT with polyethylene glycol [PEG]) on the metabolome during sainfoin ensiling. In total, 510 metabolites were identified after 60 d of sainfoin ensiling, with 33 metabolites were annotated in the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Among those metabolites, phospholipids were the most abundant (72.7% of total 33 metabolites). In addition, 10 up-regulated and 23 down-regulated metabolites, respectively, were identified in the PEG treated group when compared with the control group, after 60 d of ensiling (p \u3c 0.05). Pediococcus (correlated with 20 metabolites, R2 \u3e 0.88, p\u3c 0.05) and Lactobacillus (correlated with 16 metabolites, R2 \u3e 0.88, p \u3c 0.05) were the bacteria most correlated with metabolites. The results suggest antagonistic effects between Lactobacillus and Pediococcus occur during ensiling. The proteolysis decreased partly due to CT inhibiting Pediococcus activity during ensiling, with Pediococcus being significantly and positively correlated with dopamine after 60 d of ensiling (R2=0.8857, p \u3c 0.05)

Effects of Condensed Tannins on Bacterial and Fungal Communities during Aerobic Exposure of Sainfoin Silage

Background: Sainfoin is a forage legume that is widely distributed around the world and is beneficial for animals owing to the characteristics of its condensed tannins (CTs), which, from certain plants, can prolong the aerobic stability of silage. Methods: The present study investigated whether sainfoin CTs can prolong aerobic stability by adding polyethylene glycol (PEG) to inactivate CT activity in the silage system. Results: The results showed that aerobic stability increased under the PEG treatment (p p Lactobacillus was lower in the PEG-treated group (65.01% vs. 75.01% in the control; p Pediococcus was higher in the PEG-treated group compared with the control (10.9% vs. 4.49%, respectively; p Apiotrichum and Aspergillus were lower in the control than in the PEG-treated group after 7 d of aerobic exposure. Conclusions: The results suggested that sainfoin CTs decreased aerobic stability, but could inhibit certain bacteria and fungi, such as Pediococcus and Apiotrichum, and preserve the protein content during the aerobic exposure of silage

Effects of Nitrogen and Phosphorus Fertilization on Photosynthetic Properties of Leaves and Agronomic Characters of Alfalfa over Three Consecutive Years

The present study aimed to investigate the nitrogen (N) and phosphorus (P) fertilization of continuous addition effects plant biomass, the physiological properties of leaves and the antioxi-dant enzyme activities of alfalfa (Medicago sativa L) in the northern Xinjiang region; including the no fertilization (CK), nitrogen fertilization (N, 120 kg·ha−1), phosphorus fertilization (with low amount of N) (P, 100 kg·ha−1 P and 23.5 kg·ha−1 N) and combined nitrogen and phosphorus fertilization (NP, 120 kg·ha−1 N and 100 kg·ha−1 P) on the K well supplied soil. After three consecutive years of the supply of N and P fertilization, samples were taken at the first flowering of alfalfa (four clippings in the total year) to determine its pigment concentration, stomatal aperture, antioxidant enzyme activity and hay yield. The results showed that NP fertilization promoted growth with a higher number of branches and hay yield of alfalfa, while N or P fertilization alone had a positive effect on the growth of alfalfa. However, P fertilization significantly increased the carotenoid (Car) content at the early flowering stage of alfalfa leaves (during four clippings) (p < 0.05), In addition, NP ferti-lization enhanced stomatal aperture, increased the antioxidant enzyme activity and decreased the oxidized substance at the early flowering stage of alfalfa leaves. The results showed that a N and P balance rather than an absolute amount of either enhanced the growth of alfalfa, and N or P fertili-zation affects physiological traits differently. We propose that NP fertilization increases the nutri-tional characteristics and physiological characteristics, enhancing the adaptive capacity of alfalfa and making it better adapted to external environmental changes

Study on Phyllosphere Microbial Community of Nettle Leaf during Different Seasons

Nettle (Urtica cannabina) is an excellent feed resource widely distributed worldwide. Phyllosphere microbes are important as they have living conditions similar to those of the above-ground parts of host plants. Exploring amino acids (AA) and microorganisms can further understand the growth of plants in different seasons. The present study investigated the content of AA and phyllosphere microbes’ structure of nettle plants in different seasons. The results found that AA contents varied significantly with the season, such as alanine, aspartate, cysteine, glutamate, glycine, and methionine contents decreased significantly from spring to winter (p p U. cannabina mainly participated in metabolism. Pathogenic fungi were relatively high in autumn. The present study reveals the influence of seasonal change on the phyllosphere microbial community in U. cannabina

Effect of Intrinsic Tannins on the Fermentation Quality and Associated with the Bacterial and Fungal Community of Sainfoin Silage

Sainfoin (Onobrychis viciifolia) is rich in condensed tannins (CT). CT function includes inhibiting bacterial and fungi activity during the ensiling process. We used polyethylene glycol (PEG) to deactivate tannin activity to find out the effects of CT. The results show that the addition of PEG increased dry-matter loss (8.32% vs. 14.15%, on a dry-matter basis) after 60 d of ensiling, and also increased lactic acid (10.90% vs. 15.90%, on a dry-matter basis) and acetic-acid content (7.32% vs. 13.85%, on a dry-matter basis) after 30 d of ensiling. The PEG-treated group increased its Pediococcus relative abundance (0.37–3.38% vs. 7.82–23.5%,) during the ensiling process, increased its Gibellulopsis relative abundance after 3 d of ensiling (5.96% vs. 19.52%), increased its Vishniacozyma relative abundance after 3 d and 7 d of ensiling (2.36% vs. 17.02%, 3.65% vs. 17.17%), and increased its Aspergillus relative abundance after 7 d, 14 d and 60 d of ensiling (0.28% vs. 1.32%, 0.49% vs. 2.84% and 1.74% vs. 7.56%). However, the PEG-treated group decreased its Alternaria relative abundance during entire ensiling process (14.00–25.21% vs. 3.33–7.49%). These results suggest that condensed tannins inhibit lactic-acid bacteria fermentation though reducing Pediococcus activity, and inhibiting fungi activity depending on different strains

Biodegradation of Free Gossypol by Helicoverpa armigera Carboxylesterase Expressed in Pichia pastoris

Gossypol is a polyphenolic toxic secondary metabolite derived from cotton. Free gossypol in cotton meal is remarkably harmful to animals. Furthermore, microbial degradation of gossypol produces metabolites that reduce feed quality. We adopted an enzymatic method to degrade free gossypol safely and effectively. We cloned the gene cce001a encoding carboxylesterase (CarE) into pPICZαA and transformed it into Pichia pastoris GS115. The target protein was successfully obtained, and CarE CCE001a could effectively degrade free gossypol with a degradation rate of 89%. When esterase was added, the exposed toxic groups of gossypol reacted with different amino acids and amines to form bound gossypol, generating substances with (M + H) m/z ratios of 560.15, 600.25, and 713.46. The molecular formula was C27H28O13, C34H36N2O6, and C47H59N3O3. The observed instability of the hydroxyl groups caused the substitution and shedding of the group, forming a substance with m/z of 488.26 and molecular formula C31H36O5. These properties render the CarE CCE001a a valid candidate for the detoxification of cotton meal. Furthermore, the findings help elucidate the degradation process of gossypol in vitro

Integrated Design of a Functional Composite Electrolyte and Cathode for All-Solid-State Li Metal Batteries

Solid composite electrolytes exhibit tremendous potential for practical all-solid-state lithium metal batteries (ASSLMBs), whereas the interfacial contact between cathode and electrolyte remains a long-standing problem. Herein, we demonstrate an integrated design of a double-layer functional composite electrolyte and cathode (ID-FCC), which effectively improves interfacial contact and increases cycle stability. One composite electrolyte layer, PVDFLiFSI@LLZNTO (PL1@L), comes into contact with the LLZNTO (Li6.5La3Zr1.5Nb0.4Ta0.1O12)-containing cathode, while the other layer, PEOLiTFSI@LLZNTO (PL2@L) with a Li anode, is introduced in each. Such a design establishes a continuous network for the transport of Li+ on the interface, and includes the advantages of both PEO and PVDF for improving stability with the electrodes. The Li symmetric cells Li/PL2@L-PL1@L-PL2@L/Li steadily cycled for more than 3800 h under the current density of 0.05 mA cm−2 at 60 °C. Outstandingly, the all-solid-state batteries of LiFePO4-ID-FCC/Li showed an initial specific capacity of 161.5 mA h g−1 at 60 °C, demonstrating a remaining capacity ratio of 56.1% after 1000 cycles at 0.1 C and 74.5% after 400 cycles at 0.5 C, respectively. This work provides an effective strategy for solid-state electrolyte and interface design towards ASSLMBs with high electrochemical performance

Table_4_Chemical characteristics, antioxidant capacity, bacterial community, and metabolite composition of mulberry silage ensiling with lactic acid bacteria.DOCX

Mulberry has high crude protein and biologically active compounds but is difficult to be ensiled due to the lack of adequate epiphytic LAB. This study aimed to investigate the effects of inoculation with Lactiplantibacillus plantarum and Pediococcus pentosaceus isolated from mulberry with higher antioxidant capacity alone or in combination with Streptococcus bovis on chemical characteristics, antioxidant capacity, bacterial community, and metabolite composition of mulberry silage. The results showed that all inoculation groups had higher dry matter and lower pH than the control group, particularly in LP (dry matter, DM, 32.03% and pH = 4.44) and LP_PP_SB (DM, 31.68% and pH = 4.26) after 60 days of ensiling. Ammonia nitrogen (AN) content was the lowest in both LP_SB and LP_PP_SB groups, which were 1.86 g/kg FM and 1.05 g/kg FM, respectively, (P < 0.05). Only the LP_PP_SB group showed increased polyunsaturated fatty acids (PUFA, 1.2851 g/kg DM, P < 0.05) than the control group. Ferric-reducing antioxidant power (FRAP) values were increased in all inoculation-treated groups compared with the control group (P < 0.05). 2,2-Diphenyl-1-picrylhydrazyl (DDPH), 3-ethylbenzothiazoline-6-sulfonic acid (ABTS), and FRAP exhibited the highest levels in the LP_PP- and LP_PP_SB-treated groups. Enterobacter was dominant in both the control and SB-treated groups, and the relative abundance was 41.18% and 32.35%, respectively (P < 0.05). The relative abundance of Lactiplantibacillus was higher in the LP-, LP_PP-, and LP_SB-treated groups (81.84%−82.69%). Relative abundance of Pediococcus was higher in the PP-, PP_SB-, and LP_PP_SB-treated groups (74.27%−85.27%). Untargeted metabolomics analysis results showed that five flavonoids (apigenin, eriodictyol, quercetin-3-glucoside, rutin, and kaempferol-3-O-rutinoside)were upregulated in all inoculation groups (except for the SB-treated groups). Among them, eriodictyol was both positively correlated with ABTS and FRAP and also showed the highest relative abundance in the LP_PP- and LP_PP_SB-treated groups. To the best of our knowledge, this study was the first to investigate the relationship between inoculants of epiphytic lactic acid bacteria and antioxidant capacity by 16s rRNA Illumina sequencing technology and untargeted metabolomics analysis, respectively. Consequently, inoculated L. plantarum, P. pentosaceus alone, respectively, or in combination with S. bovis increased the relative abundance of Lactiplantibacillus and Pediococcus and decreased the relative abundance of Enterobacter, particularly in the LP_PP_SB-treated group. In addition, inoculants could increase the relative abundance of five flavonoids (apigenin, eriodictyol, quercetin-3-glucoside, rutin, and kaempferol-3-O-rutinoside), especially eriodictyol to improve the antioxidant capacity of mulberry silage.</p

Image_2_Chemical characteristics, antioxidant capacity, bacterial community, and metabolite composition of mulberry silage ensiling with lactic acid bacteria.TIF

Mulberry has high crude protein and biologically active compounds but is difficult to be ensiled due to the lack of adequate epiphytic LAB. This study aimed to investigate the effects of inoculation with Lactiplantibacillus plantarum and Pediococcus pentosaceus isolated from mulberry with higher antioxidant capacity alone or in combination with Streptococcus bovis on chemical characteristics, antioxidant capacity, bacterial community, and metabolite composition of mulberry silage. The results showed that all inoculation groups had higher dry matter and lower pH than the control group, particularly in LP (dry matter, DM, 32.03% and pH = 4.44) and LP_PP_SB (DM, 31.68% and pH = 4.26) after 60 days of ensiling. Ammonia nitrogen (AN) content was the lowest in both LP_SB and LP_PP_SB groups, which were 1.86 g/kg FM and 1.05 g/kg FM, respectively, (P < 0.05). Only the LP_PP_SB group showed increased polyunsaturated fatty acids (PUFA, 1.2851 g/kg DM, P < 0.05) than the control group. Ferric-reducing antioxidant power (FRAP) values were increased in all inoculation-treated groups compared with the control group (P < 0.05). 2,2-Diphenyl-1-picrylhydrazyl (DDPH), 3-ethylbenzothiazoline-6-sulfonic acid (ABTS), and FRAP exhibited the highest levels in the LP_PP- and LP_PP_SB-treated groups. Enterobacter was dominant in both the control and SB-treated groups, and the relative abundance was 41.18% and 32.35%, respectively (P < 0.05). The relative abundance of Lactiplantibacillus was higher in the LP-, LP_PP-, and LP_SB-treated groups (81.84%−82.69%). Relative abundance of Pediococcus was higher in the PP-, PP_SB-, and LP_PP_SB-treated groups (74.27%−85.27%). Untargeted metabolomics analysis results showed that five flavonoids (apigenin, eriodictyol, quercetin-3-glucoside, rutin, and kaempferol-3-O-rutinoside)were upregulated in all inoculation groups (except for the SB-treated groups). Among them, eriodictyol was both positively correlated with ABTS and FRAP and also showed the highest relative abundance in the LP_PP- and LP_PP_SB-treated groups. To the best of our knowledge, this study was the first to investigate the relationship between inoculants of epiphytic lactic acid bacteria and antioxidant capacity by 16s rRNA Illumina sequencing technology and untargeted metabolomics analysis, respectively. Consequently, inoculated L. plantarum, P. pentosaceus alone, respectively, or in combination with S. bovis increased the relative abundance of Lactiplantibacillus and Pediococcus and decreased the relative abundance of Enterobacter, particularly in the LP_PP_SB-treated group. In addition, inoculants could increase the relative abundance of five flavonoids (apigenin, eriodictyol, quercetin-3-glucoside, rutin, and kaempferol-3-O-rutinoside), especially eriodictyol to improve the antioxidant capacity of mulberry silage.</p