Probiotic effect of ferulic acid esterase-producing Lactobacillus plantarum inoculated alfalfa silage on digestion,antioxidant,and immunity status of lactating dairy goats

A feeding experiment was conducted to determine the effects of inoculating alfalfa silage with a ferulic acid esterase-producing inoculum on feed digestibility,rumen fermentation,antioxidant,and immunity status of lactating dairy goats.Twenty dairy goats were distributed into 2 experimental groups consisting of control diet(Lp MTD/1,including Lactobacillus plantarum MTD/1 inoculated silage)against diet containing silage treated with ferulic acid esterase-producing L.plantarum A1(Lp A1).Alfalfa silage inoculated with a ferulic acid esterase-producing Lp A1 had better fermentation quality than the Lp MTD/1inoculation.The application of Lp A1 improved silage antioxidant capacity as indicated by greater total antioxidant capacity(T-AOC),superoxide dismutase(SOD)and glutathion peroxidase(GSH-Px)activities in Lp A1 treated silage versus Lp MTD/1 treatment.Compared with Lp MTD/1 treated group,inoculation of silage with Lp A1 increased apparent total tract digestibility of dietary dry matter,organic matter and crude protein,and ruminal concentrations of total volatile fatty acids,acetate,propionate and isobutyrate as well.The results of current study also demonstrated improved antioxidant capacity and immune performance of dairy goats with Lp A1 inoculation.Feeding Lp A1-treated silage increased dairy goats’serum antioxidase activity,such as T-AOC,SOD,GSH-Px and catalase,and the serum concentration of immunoglobulin A,while decreased tumor necrosis factor a,interleukin(IL)-2 and IL-6.In addition,compared with Lp MTD/1,diet containing alfalfa silage inoculated with Lp A1 endowed dairy goats’milk with greater fat and protein contents,improved dairy goat milk quality without affecting feed efficiency.

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.

Breakdown of Corn Fiber by a Metagenomic Ferulolyl Esterase in Combination with Glycosyl Hydrolases

A feruloyl esterase (FAE-C6) gene of 957 bp was isolated from rumen microbial metagenome, subcloned into pET32b vector, and expressed in Escherichia coli. The enzyme purified in active form, consisted of 319 amino acid residues, with a molecular weight of 43.7 based on SDS-PAGE. Homology modeling showed that the FAE contained the catalytic triad composed of Ser154-Asp263His295 and a classical Gly-X-Ser154-X-Gly nucleophile motif commonly found in esterases. The FAE-C6 was characterized using corn fiber as substrate. Its combining action with glycoside hydrolases (C, X, A) individually and in various combinations was studied with focus on the difference in the effects on FA and sugar release. Glycoside hydrolases with endo-xylanase included in the enzyme mixture showed significant impact on increasing the FA yield. For the release of sugar, FAE enhanced the yield in all hydrolase combinations moderately and endo-xylanase was not the key factor in the enzyme formulation.

Severe chronic diarrhea and weight loss in cholesteryl ester storage disease: A case report

AIM: An inherited deficiency of human lysosomal acid lipase (LAL)results in the rare conditions of Wolman disease and cholesteryl ester storage disease (CESD). We want to present the rare case of CESD in an adult.METHODS: We report about an adult female patient with severe chronic diarrhea and weight loss as a consequence of CESD. Clinical examination revealed signs of malabsorption and slightly elevated liver enzymes.RESULTS: Histopathologic changes in the liver tissue and DNA sequence analysis confirmed the diagnosis of CESD due to homozygosity for the most common CESD mutation,a G934A splice site defect encoded by exon 8 of the lysosomal acid lipase (LIPA) gene.CONCLUSION: It is the first case in the literature with diarrhea as a putative symptom of CESD in adult patients.