Targeted Metabolomics Analysis Reveals that Dietary Supranutritional Selenium Regulates Sugar and Acylcarnitine Metabolism Homeostasis in Pig Liver.

BACKGROUND: The association between high selenium (Se) intake and metabolic disorders such as type 2 diabetes has raised great concern, but the underlying mechanism remains unclear. OBJECTIVE: Through targeted metabolomics analysis, we examined the liver sugar and acylcarnitine metabolism responses to supranutritional selenomethionine (SeMet) supplementation in pigs. METHODS: Thirty-six castrated male pigs (Duroc-Landrace-Yorkshire, 62.0 ± 3.3 kg) were fed SeMet adequate (Se-A, 0.25 mg Se/kg) or SeMet supranutritional (Se-S, 2.5 mg Se/kg) diets for 60 d. The Se concentration, biochemical, gene expression, enzyme activity, and energy-targeted metabolite profiles were analyzed. RESULTS: The Se-S group had greater fasting serum concentrations of glucose (1.9-fold), insulin (1.4-fold), and free fatty acids (FFAs,1.3-fold) relative to the Se-A group (P < 0.05). The liver total Se concentration was 4.2-fold that of the Se-A group in the Se-S group (P < 0.05), but expression of most selenoprotein genes and selenoenzyme activity did not differ between the 2 groups. Seven of 27 targeted sugar metabolites and 4 of 21 acylcarnitine metabolites significantly changed in response to high SeMet (P < 0.05). High SeMet supplementation significantly upregulated phosphoenolpyruvate carboxy kinase (PEPCK) activity by 64.4% and decreased hexokinase and succinate dehydrogenase (SDH) activity by 46.5-56.7% (P < 0.05). The relative contents of glucose, dihydroxyacetone phosphate, α-ketoglutarate, fumarate, malate, erythrose-4-phosphate, and sedoheptulose-7-phosphate in the Se-S group were 21.1-360% greater than those in the Se-A group (P < 0.05). The expression of fatty acid synthase (FASN) and the relative contents of carnitine, hexanoyl-carnitine, decanoyl-carnitine, and tetradecanoyl-carnitine in the Se-S group were 35-97% higher than those in the Se-A group (P < 0.05). CONCLUSIONS: Dietary high SeMet-induced hyperglycemia and hyperinsulinemia were associated with suppression of sugar metabolism and elevation of lipid synthesis in pig livers. Our research provides novel insights into high SeMet intake-induced type 2 diabetes.

Metabolism and biological activity of α-tocopherol derived from vitamin E-enriched transgenic maize in broilers.

BACKGROUND: The aim of this study was to investigate the metabolism of α-tocopherol derived from vitamin E-enriched transgenic maize (VER) and its effects on antioxidant and immune functions in broilers aged 1-42 days. A total of 360 1-day-old male broilers were randomly divided into three groups containing six replicates with 20 broilers per replicate. The negative control (NC) group and the positive control (PC) group were given non-GM maize and non-GM maize plus exogenous vitamin E (VE), respectively, and the VER group was given VER, replacing the non-GM maize given to the NC group. Between days 1 and 21 and days 22 and 42, VE levels were 4.38 and 4.63 mg kg-1 in the NC group, and 14.11 and 14.91 mg kg-1 in the PC and VER group, respectively. RESULTS: The results showed that α-tocopherol from both VER and additives increased α-tocopherol transfer protein and cytochrome P450 concentrations. Serum α-tocopherol and α-tocopherylquinone levels of broilers in the PC and VER groups were also significantly higher than those in the NC group (P < 0.05). Compared with the NC group, broilers in both groups that received α-tocopherol had reduced NF-κB p65 concentrations, significantly decreased serum prostaglandin E2 , interleukin-6, malondialdehyde, and hydrogen peroxide levels (P < 0.05), and significantly increased glutathione, glutathione peroxidase, and total antioxidant capacity (P < 0.05). CONCLUSION: In summary, both VER and non-GM maize fortified with exogenous VE showed similar effects on broilers, indicating that the α-tocopherol in VER has sufficient biological activity. © 2020 Society of Chemical Industry.

Effect of Replacing in-Feed Antibiotic Growth Promoters with a Combination of Egg Immunoglobulins and Phytomolecules on the Performance, Serum Immunity, and Intestinal Health of Weaned Pigs Challenged with Escherichia coli K88.

The study was conducted to investigate the effects of replacing antibiotic growth promoters (AGPs) with an egg immunoglobulin (IgY) combined with phytomolecules (PM) on the growth rate, serum immunity, and intestinal health of weaned pigs challenged with Escherichia coli K88 (E. coli K88). A total of 192 piglets were weaned at 28 days old with an average weight of 7.29 (± 0.04) kg. They were randomly divided into four treatments containing eight replicates with six piglets per replicate. The treatment groups were NC and PC fed a basal diet, AGP fed a basal diet supplemented with 75 mg/kg chlortetracycline, 50 mg/kg oxytetracycline calcium, and 40 mg/kg zinc bacitracin, IPM fed a basal diet supplemented with IgY at dose of 2.5 g/kg and 1.0 g/kg and PM at dose of 300 mg/kg and 150 mg/kg during days 1 to 17 and 18 to 42, respectively. On days 7 to 9 of the experiment, piglets in the PC, AGP, and IPM groups were orally challenged with 20 mL E. coli K88 (109 CFU/mL), while piglets in the NC group were challenged with 20 mL medium without E. coli K88. The E. coli K88 challenge model was successful as the incidence of post-weaning diarrhea (PWD) of piglets challenged with E. coli K88 was significantly higher than that of those unchallenged piglets during the challenge time (days 7 to 9) and days 1 to 7 of post-challenge (p < 0.05). A diet with combinations of IgY and PM and AGPs significantly decreased the incidence of PWD during the challenge time and days 1 to 7 of post-challenge (p < 0.05) compared to the PC group and significantly improved the ratio of feed to weight gain (F:G) during days 1 to 17 of the experiment compared to the NC and PC groups (p < 0.05). In comparison with the PC group, piglets in the IPM group had significantly higher serum levels of IgA, IgG, and IgM (p < 0.05), but lower serum IL-1ß on day 17 of experiement (p < 0.05). Besides, diet supplementation with AGP significantly decreased serum IL-1ß, IL-6, and TNF-α on days 17 and 42 (p < 0.05) with comparison to the PC group. Piglets in the IPM group showed a significantly lower level of fecal coliforms (p < 0.05), but a higher villus height of jejunum and ileum and higher ratio of villus height to crypt depth of duodenum and jejunum (p < 0.05) than those piglets in the PC group. In summary, diet supplementation with a mixture of IgY and PM decreased the incidence of PWD and coliforms, increased feed conversion ratio, and improved intestinal histology and immune function.

Effects of Dietary Supplementation with Clostridium butyricum on Growth Performance, Serum Immunity, Intestinal Morphology, and Microbiota as an Antibiotic Alternative in Weaned Piglets.

This study investigated the effects of Clostridium butyricum (C. butyricum) use on growth performance, serum immunity, intestinal morphology, and microbiota as an antibiotic alternative in weaned piglets. Over the course of 28 days, 120 piglets were allocated to four treatments with six replicates of five piglets each. The treatments were: CON (basal diet); AGP (basal diet supplemented with 0.075 g/kg chlortetracycline, 0.055 g/kg kitasamycin, and 0.01 g/kg virginiamycin); CBN (basal diet supplemented with normal dosage of 2.5 × 108 CFU/kg C. butyricum); and CBH (basal diet supplemented with high dosage of 2.5 × 109 CFU/kg C. butyricum). Body weight (BW) and feed consumption were recorded at the beginning and on days 14 and 28 of the experiment, and representative feed samples and fresh feces were collected from each pen between days 26 and 28. Average fecal score of diarrhea was visually assessed each morning during the experimental period. On the morning of days 14 and 28, blood samples were collected to prepare serum for immune and antioxidant parameters measurement. One male piglet close to the average group BW was selected from each replicate and was slaughtered on day 21 of the experiment. Intestinal crypt villi, and colonic microbiota and its metabolites short-chain fatty acids were measured. Compared to the CON group, the CBN and AGP groups significantly decreased (p < 0.05) the ratio of feed to weight gain by 8.86% and 8.37% between days 1 and 14, 3.96% and 13.36% between days 15 and 28, 5.47% and 11.44% between days 1 and 28. Dietary treatment with C. butyricum and AGPs significantly decreased the average fecal score during the experimental period (p < 0.05). The apparent total tract digestibility of dry matter, organic matter, and total carbohydrates in the CBH group were higher respectively at 3.27%, 2.90%, and 2.97%, than those in the CON or AGP groups (p < 0.05). Compared to the CON group, the CBH group significantly increased short-chain fatty acids in colon and villus height in the jejunum (p < 0.05). The CBN group had higher serum levels of immunoglobulins, interleukin 2 (IL-2), and glutathione peroxidase (GSH-PX) activity, but lower serum levels of IL-1ß and IL-6, and a lower aspartate aminotransferase (AST), alkaline phosphatase (ALP), and gamma-glutamyl transpeptidase (γ-GT) activity (p < 0.05), while compared to the CON group. Dietary treatment with C. butyricum significantly increased the relative abundance of Streptococcus and Bifidobacterium (p < 0.05). In summary, diet with C. butyricum increased the growth performance and benefited the health of weaned piglets.

Effect of Different Selenium Sources on Growth Performance, Tissue Selenium Content, Meat Quality, and Selenoprotein Gene Expression in Finishing Pigs.

Se-methylselenocysteine (MeSeCys) is a natural organic selenium (Se) supplement. However, its effects on animal nutrition are poorly understood. This study compared the effects of sodium selenite (SeNa), MeSeCys, and selenomethionine (SeMet) on immune function, tissue Se concentration, meat quality, and selenoprotein gene expression in pigs. A total of 72 finishing pigs were divided into four groups, which received a basal diet (BD, 0.1 mg Se/kg) without Se supplementation or one supplemented with SeNa, MeSeCys, or SeMet at a concentration of 0.25 mg Se/kg. Organic Se supplementation significantly increased the immune globulin A (IgA), IgG, and IgM serum levels compared with BD and SeNa groups (P < 0.05). There were no statistically significant differences in growth performance among the four groups. SeMet was more efficient in increasing Se concentrations in the heart, muscle, and liver than MeSeCys and SeNa (P < 0.05), while no statistically significant differences were observed between MeSeCys and SeNa. Se supplementation significantly decreased the pressing muscle loss compared with the BD group (P < 0.05). Meat color and pH were not significantly affected. Se supplement effects on liver selenoprotein gene mRNA level enhancement were ranked as follows: MeSeCys > SeMet > SeNa (P < 0.05). In muscle tissues, only the SELENOW mRNA level was significantly increased by the MeSeCys and SeMet treatment, compared with the SeNa group. In conclusion, SeMet was more efficient in increasing Se concentrations than MeSeCys and SeNa in pigs, while MeSeCys was more efficient in enhancing selenoprotein gene expression than SeMet and SeNa.

Selenium deficiency-induced redox imbalance leads to metabolic reprogramming and inflammation in the liver.

Selenium (Se) intake disequilibrium is associated with many human diseases (e.g., Keshan disease and type 2 diabetes). To understand the mechanism of Se deficiency-induced hepatic pathogenesis, a pure line pig model was established by feeding a diet with either 0.07 mg/kg Se or 0.3 mg/kg Se for 16 weeks. The hepatic metabolome, lipidome, global proteome, and whole transcriptome were analyzed. Se deficiency causes a redox imbalance via regulation of selenoproteins at both the mRNA and protein level, and blocks the glutathione anti-oxidant system along with enhanced glutathione synthesis and catabolism. The Warburg effect was observed by enhanced activation of the glycolysis and phosphate pentose pathways. The tricarboxylic acid cycle was dysfunctional since the preliminary metabolites decreased and shifted from using glycolysis origin substrates to a glutamine catabolism-preferred metabolic mode. The reprogrammed central carbon metabolism induced widely restrained lipid synthesis. In addition, a Se deficiency initiated inflammation by activating the NF-κB pathway through multiple mechanisms. These results identified the potential metabolic vulnerability of the liver in response to a Se deficiency-induced redox imbalance and possible therapeutic or intervention targets.

Development and application of a HPLC-ICP-MS method to determine selenium speciation in muscle of pigs treated with different selenium supplements.

Dietary selenium deficiency is recognized as a global problem. Pork is the most widely consumed meat throughout the world and an important source of selenium for humans. In this study, a reliable approach was developed for analyzing selenium and its speciation in the muscles of pigs after different selenium treatments. The selenium source deposition efficiency was ranked as: selenomethionine > methylselenocysteine > selenite, and the muscle selenium content had a dose effect with selenomethionine supplementation. In total, four species of selenium were detected in the muscles of pigs and the distributions of these selenium species were greatly affected by the dietary selenium supplementation forms and levels. Selenomethionine (>70% of total selenium) and selenocystine (>11%) were the major selenium species, followed by methylselenocysteine and selenourea. Therefore, selenium-enriched pork produced from selenomethionine is a good source for improving human dietary selenium intake.

Application of isotopic and elemental fingerprints in identifying the geographical origin of goat milk in China.

In the study, the samples of goat milk, feed, soil and drinking water were firstly collected from nine farms of three provinces in China, the stable isotopic ratios (δ13C, δ15N, δ2H and δ18O) in casein, lipid of milk, feed, and drinking water and the contents of 11 elements in milk and soil were determined. Statistical analysis indicated that except the content of Zn, other variables in milk showed significant differences among two or three regions. Significant correlations were found for δ13C between casein and lipid (r = 0.953, P < 0.05) and for δ2H between casein and milk water (r = 0.816, P < 0.05). The δ2H and δ18O in casein and milk water were identical to that in corresponding drinking water. Finally, seven key variables (δ15Ncasein, δ18Ocasein, Ni, Se, Rb, Sr and Ba) were selected by stepwise canonical discriminant analysis and the correct classification rate reached 100%.

Metabolic Characterization of Dairy Cows Treated with Gossypol by Blood Biochemistry and Body Fluid Untargeted Metabolome Analyses.

To characterize the metabolic disorders of dairy cows treated with gossypol, 12 dairy cows were assigned to either a control group or a treatment group that was fed 1000 mg of gossypol per kilogram of dry matter feed for 28 days. Milk quality was adversely affected, as both milk protein and lactose levels were significantly decreased in the gossypol-treated group (3.40% vs 3.16%, P = 0.044; 5.15% vs 4.91%, P = 0.027; respectively). Plasma samples revealed increases in alanine aminotransferase (P = 0.092), choline esterase (P = 0.02), and glutathione transferase (P = 0.0005) and decreases in glucose (P = 0.076) in the gossypol-treated group. Mass spectrometry based comparative metabolomic analyses showed reduced concentrations of the gluconeogenesis precursor l-glutamine (P = 0.047), with significant decreases (P < 0.05) in plasma l-lysine, l-threonine, and homoserine levels after gossypol treatment. HDL-C and LDL-C levels in the gossypol-treated group were increased (P = 0.044) and decreased (P = 0.023), respectively. These results demonstrate that gossypol induced oxidative stress and hepatotoxicity; reduced peripheral lipid metabolism, and enhanced hepatic lipid accumulation; decreased amino acid bioavailability and milk protein synthesis; and decreased gluconeogenesis and milk lactose in dairy cows.

Multi-omics analyses of red blood cell reveal antioxidation mechanisms associated with hemolytic toxicity of gossypol.

Gossypol is an antiproliferative drug with limited use due to its hemolytic toxicity. In this study, accelerated hemolysis was observed in the cows treated with gossypol. Comparative metabolomics were used to gain responsive pathways in the red blood cell (RBC) to the treatment, which were crossly validated by parallel iTRAQ-based proteomic analysis and enzyme activity assay. We found that gossypol treatment appeared to considerably activate pentose phosphate pathway (PPP) with an increased key product of ribose-5-phosphate and the increased abundance and activity of several key enzymes such as 6-phosphogluconate dehydrogenase, flavin reductase, and ribose-phosphate pyrophesphokinase. Meanwhile, a decreased glycolysis metabolism was observed, as many input metabolites of glycolysis were reduced in the gossypol group, whereas its distal metabolites were unchanged, along with decreased abundance of triosephosphate isomerase and increased abundance of enzymes catalyzing several distal glycolytic steps. Oxidative reduction pathways were also remarkably affected as we found a decreased substrate of flavin reductase, glutathione disulfide, increased glutathione reductase activity, and increased abundance and activity of glutathione S-transferase with the increase of its catalytic product, cysteine. Our results demonstrated that glycolysis, PPP, and oxidative reduction pathways of RBC were all involved in RBC's response to the hemolytic toxicity of gossypol.