Ferulic acid reduces inflammatory response induced by radiation through Sirt1-NLRP3 pathway

Objective:A model of inflammatory damage was induced by radiation to investigate whether ferulic acid(FA)can reduce the inflammatory response through the Sirt1-NLRP3 inflammatory pathway.This will help discover radiation-protective drugs and elucidate the molecular mechanisms related to radiation-induced inflammatory damage.Methods:A mouse model of radiation-induced immunoinflammatory injury was established to verify the anti-inflammatory effects of FA in vivo.C57BL/6J mice were randomly divided into six groups,and 5 Gy whole-body irradiation was used for modeling.Mice were administered a gastric solvent,amifostine,or 25,50,or 100 mg/kg FA daily for 12 days,consecutively,before irradiation.The serum of mice was collected 24 hour after irradiation to observe the content of inflammatory factors interleukin(IL)-1β,IL-18,IL-6,and tumor necrosis factor(TNF)-α.The spleen and thymus tissues of mice were weighed and the organ index was calculated for pathological testing and immunofluorescence detection.Results:FA reduced the radiation-induced decrease in the spleen and thymus indices.FA significantly reduced the secretion of inflammatory factors in the serum and reversed the radiation-induced reduction in lymphocytes in the spleen and thymus of mice.FA activated Sirt1 and inhibited the expression of the NLRP3 inflammasome to alleviate the inflammatory response.Conclusions:FA reduced radiation-induced inflammation in animals,possibly by activating Sirt1 and reducing nucleotide oligomerization domain(NOD)-like receptor thermal protein domain associated protein 3(NLRP3)inflammasome expression,thereby reducing the secretion of inflammatory factors.

Quantitative Extraction of p-Coumaric Acid and Ferulic Acid in Different Gramineous Materials and Structural Changes of Residual Alkali Lignin

Ferulic acid(FA)and p-coumaric acid(pCA)in bagasse,wheat straw,corn straw,and corncob were extracted by alkaline hydrolysis and characterized by gas chromatography(GC)and gas chromatography-mass spectrometry(GC-MS).It was found that the FA and most of the pCA in gramineous biomass could be dissociated and released after being treated with 1 M NaOH at 100℃for 4 h.The yields of pCA/FA in bagasse,wheat straw,corn straw,and corncob determined by GC-FID are 39.8/11.5,13.7/11.0,28.0/11.0,and 35.1/14.5 mg/g,respectively.The raw materials and the treated solid residues were characterized by gel-state 2D Heteronuclear Single Quantum Coherence Nuclear Magnetic Resonance(2D HSQC NMR).It was found that only a small amount of lignin was detected in the residue after alkali treatment,indicating that the alkali treatment conditions can effectively cleave the FA and pCA.Additionally,the lignin in the alkali solution was recovered and characterized by 2D HSQC NMR.The FA was not able to be detected by NMR,whereas a small amount of pCA remained in the alkali lignin.This study reveals the structural change of residual lignins during the quantitative isolation of FA and pCA,which is essential for the selective isolation of pCA/FA and valorization of residual alkali lignin.

Integrated mass spectrometry imaging reveals spatial-metabolic alteration in diabetic cardiomyopathy and the intervention effects of ferulic acid

Diabetic cardiomyopathy(DCM)is a metabolic disease and a leading cause of heart failure among people with diabetes.Mass spectrometry imaging(MSI)is a versatile technique capable of combining the molecular specificity of mass spectrometry(MS)with the spatial information of imaging.In this study,we used MSI to visualize metabolites in the rat heart with high spatial resolution and sensitivity.We optimized the air flow-assisted desorption electrospray ionization(AFADESI)-MSI platform to detect a wide range of metabolites,and then used matrix-assisted laser desorption ionization(MALDI)-MSI for increasing metabolic coverage and improving localization resolution.AFADESI-MSI detected 214 and 149 metabolites in positive and negative analyses of rat heart sections,respectively,while MALDI-MSI detected 61 metabolites in negative analysis.Our study revealed the heterogenous metabolic profile of the heart in a DCM model,with over 105 region-specific changes in the levels of a wide range of metabolite classes,including carbohydrates,amino acids,nucleotides,and their derivatives,fatty acids,glycerol phospholipids,carnitines,and metal ions.The repeated oral administration of ferulic acid during 20 weeks significantly improved most of the metabolic disorders in the DCM model.Our findings provide novel insights into the molecular mechanisms underlying DCM and the potential of ferulic acid as a therapeutic agent for treating this condition.

Protective effects of ferulic acid against ionizing radiation-induced oxidative damage in rat lens through activating Nrf2 signal pathway

AIM: To examine the protection of ferulic acid(FA) against ionizing radiation(IR)-induced lens injury in rats, as well as the underlying mechanisms.METHODS: FA(50 mg/kg) was administered to rats for 4 consecutive days before they were given 10 Gy γ-radiation, as well as for 3 consecutive days afterward. Two weeks after radiation, the eye tissues were collected. Histological alterations were evaluated by hematoxylineosin staining. Enzyme linked immunosorbent assay(ELISA) was utilized to assess the activities of glutathione reductase(GR) and superoxide dismutase(SOD), as well as the levels of glutathione(GSH) and malondialdehyde(MDA) in the lenses. The protein and m RNA levels of Bcl-2, caspase-3, Bax, heme oxygenase-1(HO-1), and glutamatecysteine ligase catalytic subunit(GCLC) were quantified using Western blot and quantitative reverse transcription polymerase chain reaction, respectively. With nuclear extracts, the nuclear factor erythroid-2 related factor(Nrf2) protein expressions in the nuclei were also measured.RESULTS: Rats exposed to IR showed lens histological alterations which could be alleviated by FA. FA treatment reversed apoptosis-related markers in IR-induced lens, as evidenced by lower levels of Bax and caspase-3 and higher level of Bcl-2. Furthermore, IR induced oxidative damage manifested by decreased GSH level, increased MDA level, and decreased SOD and GR activities. FA boosted nuclear translocation of Nrf2 and increased the expressions of HO-1 and GCLC to inhibit oxidative stress, as evidenced by an increase in GSH, a decrease in MDA, and an increase in GR and SOD activities.CONCLUSION: FA may work well in preventing and treating IR-induced cataract through promoting the Nrf2 signal pathway to attenuate oxidative damage and cell apoptosis.

Antioxidant and Anti-inflammatory Capacity of Ferulic Acid Released from Wheat Bran by Solid-state Fermentation of Aspergillus niger

Objective A strain of Aspergillus niger(A. niger), capable of releasing bound phenolic acids from wheat bran, was isolated. This strain was identified by gene sequence identification. The antioxidant and anti-inflammatory capacity of ferulic acid released from wheat bran by this A. niger strain(FA-WB) were evaluated. Methods Molecular identification techniques based on PCR analysis of specific genomic sequences were conducted; antioxidant ability was examined using oxygen radical absorbance capacity(ORAC), cellular antioxidant activity(CAA) assays, and erythrocyte hemolysis assays. RAW264.7 cells were used as a model to detect anti-inflammatory activity. Results The filamentous fungal isolate was identified to be A. niger. ORAC and CAA assay showed that FA-WB had better antioxidant activity than that of the ferulic acid standard. The erythrocyte hemolysis assay results suggested that FA-WB could attenuate AAPH-induced oxidative stress through inhibition of reactive oxy gen species(ROS) generation. FA-WB could significantly restore the AAPH-induced increase in intracellular antioxidant enzyme activities to normal levels as well as inhibit the intracellular malondialdehyde formation. TNF-?, IL-6, and NO levels indicated that FA-WB can inhibit the inflammation induced by lipopolysaccharide(LPS). Conclusion Ferulic acid released from wheat bran by a new strain of A. niger had good anti-inflammatory activity and better antioxidant ability than standard ferulic acid.

Inhibitory Effect of Ferulic Acid on Oxidation of L-DOPA Catalyzed by Mushroom Tyrosinase

The inhibitory effect of ferulic acid on the diphenolase activity of mushroom tyrosinase and the kinetic behavior were studied with L-3,4-dihydroxyphenylalanine （L-DOPA） as substrate. The inhibitor concentration leading to 50% relative activity lost （IC50） was estimated to be 0.15 mmol·L^-1. The inhibition mechanism obtained from Lineweaver-Burk plots shows that ferulic acid is a competitive inhibitor and the inhibition of tyrosinase by ferulic acid is a reversible reaction. The equilibrium constant for ferulic acid binding with the tyrosinase was determined to be 0.25 mmol·L^-1 for diphenolase. Keywords tyrosinase, ferulic acid, kinetics, inhibition, L-DOPA, diphenolase

Pharmacokinetic and pharmacodynamic analysis of ferulic acidpuerarin-astragaloside in combination with neuroprotective in cerebral ischemia/reperfusion injury in rats

Objective:To investigate the effects of the active ingredients combined therapy on inflammatory factors interleukin 1 beta(IL-1β)and neuropeptide Y(NPY)based on pharmacodynamics in rats.Methods:The animal model was built by transient middle cerebral artery occlusion(MCAO).The method for evaluating the concentrations of the FA-Pr-AI components in rat plasma was established by using HPLC and the expression levels of IL-1βand NPY were determined by ELISA.A new mathematics method of the trend of percentage rate of change(PRC)was used to assess the correlation between pharmacokinetics(PK)and pharmacodynamics(PD).Results:FA-Pr-Al in combination reduced neurological deficits,decreased infarct volume and inhibited the expression levels of IL-1βand NPY(all P<0.05)compared with the model group.FA,Pr and Al all displayed two compartment open models in rats.Clockwise hysteresis loops were obtained by time-concentration-effect curves.IL-1βand NPY level changes in the plasma followed an opposite trend to the plasma concentration tendency after C_(max)was reached.Astragaloside's PRC value was significantly higher than those of FA and puerarin between 120 to 180 min.Conclusions:The pharmacokinetics of FA-PrAl in combination were closely related its pharmacodynamics in treating ischemia/reperfusion injury,and the components of FA-Pr-Al may have a synergistic pharmacological effect.Astragaloside may play a more pronounced role in regulating IL-1βand NPY levels compared with puerarin or FA.

Ferulic acid alleviates lipopolysaccharide-induced depression-like behavior by inhibiting inflammation and apoptosis

Objective:To identify the anti-depressive effect of ferulic acid(FA)in mice exposed to lipopolysaccharide(LPS)and explore its molecular mechanisms.Methods:The mice were divided into 5 groups as follows:Control,LPS,LPS+SP,LPS+FA,and LPS+FA+anisomycin.The LPS+FA and LPS+FA+anisomycin groups were administered with FA(100 mg/kg,i.p.)once daily continuously for 7 days,and the other groups received an equivalent volume of saline.On the 7 th day,LPS(0.1 mg/m L,i.p.)was injected in all mice except the control group 30 min after FA or saline administration.The LPS+SP and LPS+FA+anisomycin groups were intravenously administered with SP600125[c-Jun N-terminal kinase(JNK)inhibitor](100μL/site,i.v.)and anisomycin(JNK activator)(100μL/site,i.v.)15 min before LPS,respectively.The depressive behaviors were assessed by open field test,sucrose preference test,and forced swimming test at 24 h post-LPS administration.Tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)levels in plasma were measured by ELISA.The levels of phospho-JNK,TNF-α,IL-1β,Bcl-2,Bax,cytochrome c and caspase-3 were evaluated by Western blotting.Results:FA alleviated depression symptoms caused by LPS in mice,including increasing sucrose water consumption in sucrose preference test and reducing the immobility time in forced swimming test.FA could inhibit upregulated levels of phospho-JNK,TNF-α,and IL-1β.FA also markedly decreased Bax,caspase-3,and cytochrome c,and increased Bcl-2 levels.Besides,SP600125 showed neuroprotective effect similar to FA which was attenuated by anisomycin.Conclusions:FA attenuates inflammation and apoptosis by inhibiting LPS-induced activation of JNK to alleviate depressionlike behaviors.

In vitro and in vivo effects of ferulic acid on gastrointestinal motility:Inhibition of cisplatin-induced delay in gastric emptying in rats

AIM: To study the effects of ferulic acid on gastrointe-stinal motility both in vitro and in vivo.METHODS: Ferulic acid induced concentration-dependent stimulation of the basal tone of isolated guinea pig ileum (2-20 μmol/L) and isolated rat fundus (0.05-0.4 mmol/L). RESULTS: Ferulic acid significantly accelerated the gastrointestinal transit and gastric emptying in rats in a dose-dependent manner (50-200 mg/kg, po). Cisplatin (2.5-20 mg/kg, ip) induced a dose-dependent delay in gastric emptying in rats. Pretreatment with ferulic acid dose-dependently, significantly reversed the cisplatin-induced delay in gastric emptying. CONCLUSION: The endogenous prostaglandins (PGs) are involved in mediating the stimulant effects of ferulic acid. This effect of dietary ferulic acid may help improve other accompanying gastrointestinal symptoms such as abdominal discomfort and also may protect against emesis induced by cytotoxic drugs.

Biodegradation of ferulic acid by a newly isolated strain of Cupriavidus sp. B-8

As a major component of lignin and abundantly existing in softwood and hardwood, ferulic acid has been used as a lignin-related compound for lignin biodegradation study. Biodegradation of ferulic acid by Cupriavidus sp. B-8, a newly isolated strain, was studied. This strain is able to utilize a wide range of lignin-related aromatic compounds as the sole carbon and energy source, including guaiacol, veratric acid, vanillic acid, cinnamic acid, p-coumaric acid, ferulic acid, and sinapic acid. In addition, the effects of different concentrations of ferulic acid on growth of Cupriavidus sp. B-8 were studied. The growth of Cupriavidus sp. B-8 is better under the condition of lower concentration. High-performance liquid chromatography （HPLC） analysis reveals that above 95% of ferulic acid is degraded within 12 h by Cupriavidus sp. B-8. Based on identification of biodegradation intermediates and further metabolites, the biodegradation pathway of ferulic acid by Cupriavidus sp. B-8 was proposed. Ferulic acid is initially converted to 4-vinylguaiacol, and further oxidized to vanillic acid and protocatechuic acid.

Anti-insulin resistant effect of ferulic acid on high fat diet-induced obese mice

Objective: To evaluate the insulin sensitivity action of ferulic acid(FA) in skeletal muscle and hypothalamus of high-fat diet(HFD)-induced obese mice. Methods: Obese mouse model was induced by HFD(45 kcal% lard fat) for 16 weeks. After 8 weeks of HFD feeding, these obese mice were orally treated with FA at doses of 25 and 50 mg/kg/day for 8 weeks. At the end of all treatments, the epididymal fat, pancreas, skeletal muscle and hypothalamus were removed for biochemical parameter and protein expression examinations. Results: FA treatment significantly decreased leptin level in fat tissue and insulin level in pancreas(P < 0.05). Interestingly, obese mice treated with FA increased the protein expressions of insulin receptor substrate-1, phosphatidylinositol 3-kinase, and phosphorylated-protein kinase B in both muscle and brain(P < 0.05). The phosphorylations of adenosine monophosphate-activated protein kinase and acetyl-CoA carboxylase in muscle, and leptin receptor protein in hypothalamus were also increased(P < 0.05). The pancreatic islets histology showed smaller size in obese mice treated with FA compared to untreated obese mice. Conclusions: These findings indicate the beneficial effect of FA in improving insulin resistance in HFD-induced obese mice. These effects are probably mediated via modulating the insulin receptor substrate/phosphatidylinositol 3-kinase/protein kinase B or adenosine monophosphate-activated protein kinase pathways.

Synthesis, Characterization and Biological Screening of Ferulic Acid Derivatives

According to WHO, cancer is a leading cause of death worldwide, accounting for 8.2 million deaths in 2012. Among several factors involved in the pathogenesis of cancer, free radical formation followed by damage to DNA and cell protein is one of the causes. Natural plant products have gained enormous interest in the prevention or treatment of chronic diseases. Ferulic acid, like other phenolic acids (caffeic acid, sinapic acid) possess anti cancer activity. A series of ferulic esters (FE1 - FE11) and ferulic amides (FA1 - FA10) were synthesized and evaluated for their cytotoxic activity.

Evaluation of trans-ferulic acid degradation by dielectric barrier discharge plasma combined with ozone in wastewater with different water quality conditions

In this study, dielectric barrier discharge plasma and ozone(O_3) were combined to synergistically degrade trans-ferulic acid(FA), and the effect of water quality on FA degradation was studied. The results showed that 96.9% of FA was degraded after 40 min treatment by the plasma/O_3 process. FA degradation efficiency increased with the p H values. The presence of suspended solid and humic acid inhibited FA degradation. FA degradation efficiency increased as the water temperature increased to 30 °C. However, the further increase in water temperature was adverse for FA degradation. Effects of common inorganic ions on FA degradation were also investigated. The addition of Cl^- inhibited the FA degradation, whileCO_3^(2-) had both negative and positive influences on FA degradation.NO_3^- andSO_4^(2-) did not have significant effect on FA degradation. Fe^(3+)and Cu^(2+)benefited FA degradation through the Fenton-like and catalytic ozonation reactions.

Determination of Volatile Oil and Ferulic Acid in Different Parts of Wild Ferula sinkiangensis K. M. Shen and Ferula fukanensis K. M.Shen Cultivars

[Objectives] To increase the reserves distribution and planting area of Ferula resources,protect wild Ferula resources,make comparative analysis on wild and cultivated varieties of Ferula medicinal materials,and provide references for quality evaluation of Ferula herbs collected from cultivation environment. [Methods] Volatile oil determination and high performance liquid chromatography were used to measure volatile oil and ferulic acid in different parts of wild and cultivated varieties of Ferula. [Results] In the volatile oil measured from artificially pressed Ferula lipid,wild varieties in different places conformed to the pharmacopoeia standard( volatile oil content ≥10%); in cultivated varieties,F. fukanensis K. M. Shen and F. sinkiangensis K. M. shen reached the pharmacopoeia standard. According to measurement results of ferulic acid,the ferulic acid content of roots and leaves of Altay,Fukang,and Yining cultivars were higher than that of wild varieties. [Conclusions]The ferulic medicinal herbs collected from artificial cultivation environment have considerable prospect,and many parts of Ferula herbs can be used for raw materials for preparation of extracts.

Antibacterial activity and inhibition against Staphylococcus aureus NorA efflux pump by ferulic acid and its esterified derivatives

Objective:To evaluate the inhibitory activity of ferulic acid and four of its esterified derivatives(methyl,ethyl,propyl,and butyl)against resistance mechanisms in Staphylococcus aureus strains.Methods:Ferulic acid derivatives were obtained by esterification with methanol,ethanol,propanol,and butanol,and then characterized by hydrogen and carbon-13 nuclear magnetic resonance analysis.The minimum inhibitory concentrations(MIC)of ferulic acid and its esterified derivatives,ethidium bromide,and norfloxacin were obtained using the microdilution test,while the efflux pump inhibition test was conducted by examining reduction in the MICs of norfloxacin and ethidium bromide.Molecular docking was also carried out using the Schrodinger Suite 2015 molecular modeling software.A three-dimensional model of NorA efflux pump was generated using I-TASSER.The best scoring model was used as a receptor for ligand-receptor docking.Results:The methyl and butyl ester derivatives did not demonstrate significant antimicrobial activity.However,a significant synergic effect was evidenced when norfloxacin was combined with the ethyl and propyl esterified derivatives.The docking study demonstrated favorable energy of interaction between ferulate derivatives and NorA,and amino acid residues TYR57,TYR58,and LEU255 were present commonly in stabilizing all complexes.The PCA analysis corroborated the docking hypothesis that the lipophilic character and hydrogen bond interactions were the most relevant characteristics involved with NorA inhibitors.The pharmacokinetic parameters of ferulic acid derivatives showed good ADMET properties,demonstrating that they can be easily absorbed and have no effect or inhibit the cytochrome P450 enzyme complex,revealing their potential as drug candidates.Conclusions:This study provides strong evidence that the molecular basis for this activity is potentially due to the NorA efflux pump.

Effect of low-level laser irradiation(LLLI)combined with ferulic acid on the osteoblast differentiation and maturation as well as osteogenesis signaling pathway expression

Objective: To study the effect of low-level laser irradiation (LLLI) combined with ferulic acid on the osteoblast differentiation and maturation as well as osteogenesis signaling pathway expression. Methods: The cranium was collected from the SD rats born within 24 h, osteoblasts were separated, cultured and then divided into control group, ferulic acid group, LLLI group, ferulic acid + LLLI group, and the expression osteoblast differentiation markers, proliferation molecules and signaling pathway molecules were detected after continuous treatment with different conditions for three days. Results: 3 d after treatment, the Bax and Bid mRNA expression in ferulic acid group, LLLI group and ferulic acid + LLLI group of cells were significantly lower than those in control group while Bcl-2, CyclinD1, E2F, Col-I, OC and ALP mRNA expression as well as Wnt, β-catenin, Runx2, cAMP and PKA protein expression were significantly higher than those in control group;Bax and Bid mRNA expression in ferulic acid + LLLI group of cells were significantly lower than those in ferulic acid group and LLLI group while Bcl-2, CyclinD1, E2F, Col-I, OC and ALP mRNA expression as well as Wnt, β-catenin, Runx2, cAMP and PKA protein expression were significantly higher than those in ferulic acid group and LLLI group;Col-I, OC, ALP, Bax, Bid, Bcl-2, CyclinD1 and E2F mRNA expression as well as Wnt, β-catenin, Runx2, cAMP and PKA protein expression were not significantly different between ferulic acid group and LLLI group. Conclusion: Low-level laser irradiation (LLLI) combined with ferulic acid can promote osteoblast differentiation and maturation and activate osteogenesis signaling pathway.

Crystal Structure, Synthesis and Biological Activity of Ether and Ester <i>Trans</i>-Ferulic Acid Derivatives

The structures of methoxymethyl (E)-3-(4-hydroxy-3-methoxyphenyl)acrylate, 2;(E)-3-(3-methoxy-4-(methoxymethoxy)phenyl)acrylic acid, 3;methyl (E)- 3-(4-(benzyloxy)-3-methoxyphenyl)acrylate, 4;benzyl (E)-3-(4-(benzyloxy)- 3-methoxyphenyl)acrylate, 6;and (E)-3-(4-(benzyloxy)-3-methoxyphenyl)- acrylic acid, 7;were established by spectroscopic and X-ray diffraction studies. Structure 2 is a new com-pound. Compounds with free phenolic hydroxyls v.gr. methyl (E)-3-(4-hydroxy-3-methoxyphenyl)acrylate 1, 2 and ben-zyl(E)-3-(4-hydroxy-3-methoxyphenyl)acrylate 5, showed scavenging free- radical and antioxidant activity while moderate scavenging free-radical was observed in compound 3. Moderate inhibition of lipid peroxidation was observed for 7. Compound 5 exerted significant inhibition of cell growth in PC-3, K562 tumor cell lines and 4 exhibited the largest cytotoxic effect upon the K562 cell line.

Esterification of levulinic acid to n-butyl levulinate over heteropolyacid supported on acid-treated clay

Levulinic acid has been identified as a promising green, biomass-derived platform chemical. n-Butyl levulinate is used as an important intermediate having diverse applications. The present work focuses on the synthesis of n-butyl levulinate by esterification of levulinic acid with n-butanol using heteropolyacid （HPA） supported on acid-treated clay montmorillonite （K10）. 20% （w/w） dodecatungestophosphoric acid （DTPA） supported on K10 was found to be the most efficient catalyst with 97% levulinic acid conversion and 100% selectivity towards n-butyl levulinate. Effects of various process parameters were studied to examine the efficacy of 20% （w/w） DTPA/K10 for optimization of the activity.

Synthesis,Characterization and Catalytic Application of H_3PW_(12)O_(40)/MCM-48 in the Esterification of Methacrylic Acid with n-butyl Alcohol

A novel environmental friendly catalyst, H3PW12O40/MCM-48, was prepared by impregnation method. The catalysts were characterized by means of XRD and FT-IR. The synthesis of n-butyl methacrylate catalyzed by H3PW12O40/MCM-48 was studied with methacrylic acid and n-butyl alcohol as reactants. H3PW12O40/MCM-48 is an excellent catalyst for synthesizing n-butyl methacrylate and Keggin structure of H3PW12O40 kept unchanged after being impregnated on surface of the molecular sieve support. Effects of n（methacrylic acid）：n（n-butyl alcohol）, catalyst dosage, cyclohexane（water-stripped reagent） and reaction time on yields of the product were investigated. The optimum conditions have been found, that is, molar ratio of acid to alcohol is 1：1.6, mass ratio of catalyst used to the reactants is 0.5% and reaction time is 2.0 h. Under these conditions, the yield of n-butyl methacrylate can reach 93.7%.

Effects of inoculating feruloyl esterase-producing Lactiplantibacillus plantarum A1 on ensiling characteristics,in vitro ruminal fermentation and microbiota of alfalfa silage

Background Ferulic acid esterase(FAE)-secreting Lactiplantibacillus plantarum A1(Lp A1)is a promising silage inoculant due to the FAE’s ability to alter the plant cell wall structure during ensiling,an action that is expected to improve forage digestibility.However,little is known regarding the impacts of Lp A1 on rumen microbiota.Our research assessed the influences of Lp A1 in comparison to a widely adopted commercial inoculant Lp MTD/1 on alfalfa’s ensilage,in vitro rumen incubation and microbiota.Results Samples of fresh and ensiled alfalfa treated with(either Lp A1 or Lp MTD/1)or without additives(as control;CON)and ensiled for 30,60 and 90 d were used for fermentation quality,in vitro digestibility and batch culture study.Inoculants treated silage had lower(P<0.001)pH,acetic acid concentration and dry matter(DM)loss,but higher(P=0.001)lactic acid concentration than the CON during ensiling.Compared to the CON and Lp MTD/1,silage treated with Lp A1 had lower(P<0.001)aNDF,ADF,ADL,hemicellulose,and cellulose contents and higher(P<0.001)free ferulic acid concentration.Compared silage treated with Lp MTD/1,silage treated with Lp A1 had significantly(P<0.01)improved ruminal gas production and digestibility,which were equivalent to those of fresh alfalfa.Realtime PCR analysis indicated that Lp A1 inoculation improved the relative abundances of rumen’s total bacteria,fungi,Ruminococcus albus and Ruminococcus flavefaciens,while the relative abundance of methanogens was reduced by Lp MTD/1 compared with CON.Principal component analysis of rumen bacterial 16S rRNA gene amplicons showed a clear distinction between CON and inoculated treatments without noticeable distinction between Lp A1 and Lp MTD/1 treatments.Comparison analysis revealed differences in the relative abundance of some bacteria in different taxa between Lp A1 and Lp MTD/1 treatments.Silage treated with Lp A1 exhibited improved rumen fermentation characteristics due to the inoculant effects on the rumen microbial populations and bacterial community.Conclusions Our findings suggest that silage inoculation of the FAE-producing Lp A1 could be effective in improving silage quality and digestibility,and modulating the rumen fermentation to improve feed utilization.