Over-expression of a gamma-tocopherol methyltransferase gene in vitamin E pathway confers PEG-simulated drought tolerance in alfalfa

α-Tocopherol is one of the most important vitamin E components present in plant. α-Tocopherol is a potent antioxidant, which can deactivate photoproduced reactive oxygen species (ROS) and prevent lipids from oxidation when plants suffer drought stress. γ-Tocopherol methyltransferase (γ-TMT) catalyzes the formation of α-tocopherol in the tocopherol biosynthetic pathway. Our previous studies showed that over-expression of γ-TMT gene can increase the accumulation of α-tocopherol in alfalfa (Medicago sativa). However, whether these transgenic plants confer increased drought tolerance and the underlying mechanism are still unknown.This work was financially supported by Earmarked Fund for China Agriculture Research System (CARS-34), the National Natural Science Foundation of China (31872410), National Crop Germplasm Resources Center (NICGR-78), and the Agricultural Science and Technology Innovation Program (ASTIPIAS10)

Melanin: A promising source of functional food ingredient

Melanin as a natural pigment widely exists in animals, plants, fungi and bacteria. In vivo and in vitro tests have proven that melanin has a variety of health-promoting effects, such as antioxidant and immunomodulatory activities, hypoglycemic and hypolipidemic effect, liver and gastrointestinal tract protection effect. Melanin can be extracted from animal and plant by-products or produced by microbial fermentation, involving five extraction and purification methods, with their own positive and negative aspects. Melanin can be added to a variety of foods as natural colorant and preservative, with high safety and almost no toxicity. This article focuses on melanin’s source classification, health benefits, extraction technology and applications in food products to explore the potential of it as a functional food ingredient

The Effect of Epimedium Isopentenyl Flavonoids on the Broiler Gut Health Using Microbiomic and Metabolomic Analyses

Epimedium (EM), also known as barrenwort, is a traditional medicinal plant rich in isopentenyl flavonols, which have beneficial biological activities and can improve human and animal health, but its mechanism is still unclear. In this study, ultra-high-performance liquid chromatography/quadrupole-time-of-flight-mass spectrometry (UHPLC-Q-TOF/MS) and ultra-high-performance liquid chromatography triple-quadrupole mass spectrometry (UHPLC-QqQ-MS/MS) were used to analyse the main components of EM, and isopentenyl flavonols such as Epimedin A, B, and C as well as Icariin were the major components of EM. Meanwhile, broilers were selected as model animals to illuminate the mechanism of Epimedium isopentenyl flavonols (EMIE) on gut health. The results showed that supplementation with 200 mg/kg EM improved the immune response, increased cecum short-chain fatty acids (SCFAs) and lactate concentrations, and improved nutrient digestibility in broilers. In addition, 16S rRNA sequencing showed that EMIE altered the composition of cecal microbiome, increasing the relative abundance of beneficial bacteria (Candidatus Soleaferrea and Lachbospiraceae NC2004 group and Butyricioccus) and reducing that of harmful bacteria (UBA1819, Negativibacillus, and Eisenbergiella). Metabolomic analysis identified 48 differential metabolites, of which Erosnin and Tyrosyl-Tryptophan were identified as core biomarkers. Erosnin and tyrosyl-tryptophan are potential biomarkers to evaluate the effects of EMIE. This shows that EMIE may regulate the cecum microbiota through Butyricicoccus, with changes in the relative abundance of the genera Eisenbergiella and Un. Peptostreptococcaceae affecting the serum metabolite levels of the host. EMIE is an excellent health product, and dietary isopentenyl flavonols, as bioactive components, can improve health by altering the microbiota structure and the plasma metabolite profiles. This study provides the scientific basis for the future application of EM in diets

Dietary Epimedium extract supplementation improves intestinal functions and alters gut microbiota in broilers

Abstract Background Growth-promoting antibiotics have been banned by law in the livestock and poultry breeding industry in many countries. Various alternatives to antibiotics have been investigated for using in livestock. Epimedium (EM) is an herb rich in flavonoids that has many beneficial effects on animals. Therefore, this study was planned to explore the potential of EM as a new alternative antibiotic product in animal feed. Methods A total of 720 1-day-old male broilers (Arbor Acres Plus) were randomly divided into six groups and fed basal diet (normal control; NC), basal diet supplemented with antibiotic (75 mg/kg chlortetracycline; CTC), and basal diet supplemented with 100, 200, 400 or 800 mg/kg EM extract for 6 weeks (EM100, EM200, EM400 and EM800 groups). The growth performance at weeks 3 and 6 was measured. Serum, intestinal tissue and feces were collected to assay for antioxidant indexes, intestinal permeability, lactic acid and short-chain fatty acids (SCFAs) profiles, microbial composition, and expression of intestinal barrier genes. Results The average daily feed intake in CTC group at 1–21 d was significantly higher than that in the NC group, and had no statistical difference with EM groups. Compared with NC group, average daily gain in CTC and EM200 groups increased significantly at 1–21 and 1–42 d. Compared with NC group, EM200 and EM400 groups had significantly decreased levels of lipopolysaccharide and D-lactic acid in serum throughout the study. The concentrations of lactic acid, acetic acid, propionic acid, butyric acid and SCFAs in feces of birds fed 200 mg/kg EM diet were significantly higher than those fed chlortetracycline. The dietary supplementation of chlortetracycline and 200 mg/kg EM significantly increased ileal expression of SOD1, Claudin-1 and ZO-1 genes. Dietary supplemented with 200 mg/kg EM increased the relative abundances of g_NK4A214_group and Lactobacillus in the jejunal, while the relative abundances of Microbacterium, Kitasatospora, Bacteroides in the jejunal and Gallibacterium in the ileum decreased. Conclusion Supplementation with 200 mg/kg EM extract improved the composition of intestinal microbiota by regulating the core bacterial genus Lactobacillus, and increased the concentration of beneficial metabolites lactic acid and SCFAs in the flora, thereby improving the antioxidant capacity and intestinal permeability, enhancing the function of tight junction proteins. These beneficial effects improved the growth performance of broilers. Graphical Abstrac

Overexpression of alfalfa γ-tocopherol methyltransferase (γ-TMT) gene increases salt susceptibility of transgenic Arabidopsis in seed germination

As antioxidants, tocopherols deactivate reactive oxygen species and prevent lipids from oxidation in response to abiotic stresses. γ-Tocopherol methyltransferase (γ-TMT) catalyzes the conversion of γ-tocopherol into α-tocopherol which has the highest biological activity. To investigate roles of γ-TMT in seed germination under salinity stress, we heterologously overexpressed an alfalfa MsTMT gene in Arabidopsis. MsTMT transgenic seeds germinated much slower than that of Arabidopsis wild-type (WT) seeds under salt stress or exogenous abscisic acid (ABA) treatment, indicating enhanced osmotic and ABA sensitivity in transgenic seeds. Under salinity stress, expression levels of ABA biosynthesis genes (NCED4 and NCED9) and signaling genes (ABI3 and ABI5) were increased in transgenic seeds. Meanwhile, the expression of GA biosynthesis genes (GA3OX1 and GA3OX2) were repressed and that of GA signal suppressor genes RGL2 was enhanced. Moreover, overexpression of MsTMT promoted the release of seemucilage and contributed to the redistribution of pectins. Interestingly, removal of seed mucilage eliminated the difference in the initiation of seed germination between WT and transgenic lines. Taken together, MsTMT had a strong influence on the response to salinity stress in transgenic Arabidopsis during seed germination. Our results reveal a novel role of MsTMT in mediating the regulations of ABA and GA signaling in seed germination, which is also associated with mucilage release and structure. This study provides new insights into the regulatory network controlled by tocopherol biosynthesis gene in response to abiotic stress in plants.This work was supported by the grant from the Earmarked Fund for China Agriculture Research System (CARS-34), National Nature Science Foundation of China (31872410), and the Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences (ASTIP-IAS10)

A new single-cell protein from Clostridium autoethanogenum as a functional protein for largemouth bass (Micropterus salmoides).

peer reviewedClostridium autoethanogenum protein (CAP) is a new single-cell protein source originating from inactivated bacteria. An in vitro digestion experiment and an 8-wk growth experiment were conducted to evaluate the molecular weight distribution of the CAP hydrolysate, and the effects of dietary CAP levels on the growth performance, plasma parameters, hepatic and intestinal health, and the diversity of gut-adherent microbiota of largemouth bass (Micropterus salmoides). The fish (initial body weight of 47.99 ± 0.01 g) were fed diets where CAP gradually replaced 0% (CAP0), 12.5% (CAP12.5), 25% (CAP25), 37.5% (CAP37.5) and 50% (CAP50) of low-temperature steam dried anchovy fish meal (LTFM) in the diet. Results showed that the content of peptides below 1,000 Da in the CAP hydrolysate (0.56 mg/mL) was higher than that of the LTFM hydrolysate (0.48 mg/mL). Dietary CAP inclusion had no negative effect on growth performance, while whole-body lipid content significantly reduced in the CAP25 and CAP50 groups (P < 0.05). The plasma alanine aminotransferase activities and triglyceride concentrations in the CAP inclusion groups were significantly lower than those in the CAP0 group (P < 0.05). The plasma aspartate aminotransferase activity was significantly reduced in the CAP37.5 group (P < 0.05). The richness and diversity of the gut-adhesive microbiota and the proportion of Clostridium sensu stricto 12 in the CAP50 group were significantly higher than those in the CAP0 group (P < 0.05). Dietary CAP inclusion inhibited inflammatory responses by down-regulating the mRNA levels of interleukin 1β (IL1β), IL10 and transforming growth factor β1 (P < 0.05) in the liver. The mRNA levels of acetyl-CoA carboxylase 1 were significantly down-regulated in the CAP12.5, CAP25 and CAP37.5 groups (P < 0.05), while that of fatty acid synthase was significantly down-regulated in the CAP50 group (P < 0.05). These results demonstrate that dietary CAP inclusion could improve the hepatic and intestinal health of largemouth bass, and can be helpful to further develop CAP as a functional feed ingredient

Mechanism Analysis of Metabolic Fatty Liver on Largemouth Bass (Micropterus salmoides) Based on Integrated Lipidomics and Proteomics

Metabolic fatty liver disease caused by high-starch diet restricted the intensive and sustainable development of carnivorous fish such as largemouth bass. In this study, the combination liver proteomic and lipidomic approach was employed to investigate the key signaling pathways and identify the critical biomarkers of fatty liver in largemouth bass. Joint analysis of the correlated differential proteins and lipids revealed nine common metabolic pathways; it was determined that FABP1 were significantly up-regulated in terms of transporting more triglycerides into the liver, while ABCA1 and VDAC1 proteins were significantly down-regulated in terms of preventing the transport of lipids and cholesterol out of the liver, leading to triglyceride accumulation in hepatocyte, eventually resulting in metabolic fatty liver disease. The results indicate that FABP1, ABCA1 and VDAC1 could be potential biomarkers for treating metabolic fatty liver disease of largemouth bass

Comprehensive quantitation of multi-signature peptides originating from casein for the discrimination of milk from eight different animal species using LC-HRMS with stable isotope labeled peptides

Milk species adulteration has become an altering issue worldwide. In this study, a robust quantification method based on LC-HRMS for the simultaneous detection and differentiation of milk type from eight different animal species (namely: cow, water buffalo, wild yak, goat, sheep, donkey, horse, and camel) was established by detecting nine signature peptides originating from casein. The developed method was in-house validated in terms of sensitivity, accuracy, and precision. As a result, limits of quantification (LOQ) were ranging from 5 to 30 µg/L, recoveries ranged from 95.2% to 104.5%, and intra-day and inter-day variability were lower than 11.4% and 12.6%, respectively, for all the targeted peptides. Furthermore, this method was successfully applied to 46 commercial minor species’ milk, in which 15 samples were false labeling. The obtained results indicate the necessity to monitor milk species adulteration in order to protect consumers from consuming misleading labeled minor species animal’s milk