Effects of Crude Extract of Glycyrrhiza Radix and Atractylodes macrocephala on Immune and Antioxidant Capacity of SPF White Leghorn Chickens in an Oxidative Stress Model.

The natural edible characteristics of Chinese herbs have led more and more people to study them as an alternative product to antibiotics. In this study, crude extracts of Glycyrrhiza radix and Atractylodes macrocephala (abbreviated as GRAM) with glycyrrhizic acid content not less than 0.2 mg/g were selected to evaluate the effects of GRAM on the immune and antioxidant capacity of model animals. Thirty 21-day-old male Leghorn chickens were weighed and randomly assigned to one of three groups of ten animals each. The treatments comprised a control group (CON), in which saline was injected at day 31, day 33, and day 35, an LPS-treated group (LPS), in which LPS (0.5 mg/kg of BW) was injected at day 31, day 33, and day 35, and finally a GRAM and LPS-treated group, (G-L) in which a GRAM-treated diet (at GRAM 2 g/kg) was fed from day 21 to day 35 with LPS injection (0.5 mg/kg of BW) at day 31, day 33, and day 35. The results of diarrhea grade and serum antioxidant measurement showed that the LPS group had obvious diarrhea symptoms, serum ROS and MDA were significantly increased, and T-AOC was significantly decreased. The oxidative stress model of LPS was successfully established. The results of immune and antioxidant indexes showed that feeding GRAM significantly decreased levels of the pro-inflammatory factors TNF-α, IL-1ß, and IL-6 (p < 0.05) and significantly increased levels of the anti-inflammatory factors IL-4 and IL-10 and levels of the antioxidant enzymes GSH-Px and CAT (p < 0.05). GRAM resisted the influence of LPS on ileum morphology, liver, and immune organs and maintained normal index values for ileum morphology, liver, and immune organs. In summary, this study confirmed the antidiarrheal effect of GRAM, which improved the immune and antioxidant capacity of model animals by regulating inflammatory cytokine levels and antioxidant enzyme activity in poultry.

Lactiplantibacillus plantarum FRT4 attenuates high-energy low-protein diet-induced fatty liver hemorrhage syndrome in laying hens through regulating gut-liver axis.

BACKGROUND: Fatty liver hemorrhage syndrome (FLHS) becomes one of the most major factors resulting in the laying hen death for caged egg production. This study aimed to investigate the therapeutic effects of Lactiplantibacillus plantarum (Lp. plantarum) FRT4 on FLHS model in laying hen with a focus on liver lipid metabolism, and gut microbiota. RESULTS: The FLHS model of laying hens was established by feeding a high-energy low-protein (HELP) diet, and the treatment groups were fed a HELP diet supplemented with differential proportions of Lp. plantarum FRT4. The results indicated that Lp. plantarum FRT4 increased laying rate, and reduced the liver lipid accumulation by regulating lipid metabolism (lipid synthesis and transport) and improving the gut microbiota composition. Moreover, Lp. plantarum FRT4 regulated the liver glycerophospholipid metabolism. Meanwhile, "gut-liver" axis analysis showed that there was a correlation between gut microbiota and lipid metabolites. CONCLUSIONS: The results indicated that Lp. plantarum FRT4 improved the laying performance and alleviated FLHS in HELP diet-induced laying hens through regulating "gut-liver" axis. Our findings reveal that glycerophospholipid metabolism could be the underlying mechanism for the anti-FLHS effect of Lp. plantarum FRT4 and for future use of Lp. plantarum FRT4 as an excellent additive for the prevention and mitigation of FLHS in laying hens.

Optimization of Gamma-Aminobutyric Acid Production by Lactiplantibacillus plantarum FRT7 from Chinese Paocai.

Gamma-aminobutyric acid (GABA) is a widely available non-protein amino acid whose physiological importance goes beyond its role as an inhibitory neurotransmitter in mammals. The GABA synthesis ability of ten strains of Lactiplantibacillus plantarum was screened. They produced GABA ranging from 48.19 ± 3.44 to 100.75 ± 1.63 mg/L at 24 h-cultivation. Among them, Lp. plantarum FRT7 showed the highest GABA production. Therefore, FRT7 was chosen for GABA yield optimization. A one-factor-at-a-time strategy analysis of the GABA yield of FRT7 was performed, including the culture temperature, incubation time, inoculum volume, initial pH, the initial amount of monosodium glutamate (MSG), and pyridoxal 5'-phosphate (PLP) concentration, based on which the response surface methodology (RSM) was performed. After being cultured in an MRS culture medium supplemented with 3% MSG and 2 mmol/L of PLP at 40 °C with an initial pH of 7.0 for 48 h, the GABA reached a maximum yield of 1158.6 ± 21.22 mg/L. The results showed the experimental value of the GABA yield was in good agreement with the predicted values. Furthermore, the results from the RSM also indicated that the initial MSG addition, PLP concentration, and incubation time were significant variables. These results suggest that Lp. plantarum FRT7 has the potential to be a health-beneficial probiotic with commercial capabilities.

Dietary Bacillus licheniformis shapes the foregut microbiota, improving nutrient digestibility and intestinal health in broiler chickens.

Bacillus licheniformis is considered a potential alternative to antibiotic growth promoters of animal growth and health. However, the effects of Bacillus licheniformis on the foregut and hindgut microbiota, and their relationships with nutrient digestion and health, in broiler chickens remain unclear. In this study, we aimed to identify the effects of Bacillus licheniformis BCG on intestinal digestion and absorption, tight junctions, inflammation, and the fore- and hind-gut microbiota. We randomly assigned 240 1-day-old male AA broilers into three treatment groups: CT (basal diet), BCG1 (basal diet + 1.0 × 108 CFU/kg B. licheniformis BCG), and BCG2 (basal diet + 1.0 × 109 CFU/kg B. licheniformis BCG). On day 42, the jejunal and ileal chyme and mucosa were subjected to analysis of digestive enzyme activity, nutrient transporters, tight junctions, and signaling molecules associated with inflammation. The ileal and cecal chyme were subjected to microbiota analysis. Compared with the CT group, the B. licheniformis BCG group showed significantly greater jejunal and ileal α-amylase, maltase, and sucrase activity; moreover, the α-amylase activity in the BCG2 group was higher than that in the BCG1 group (P < 0.05). The transcript abundance of FABP-1 and FATP-1 in the BCG2 group was significantly greater than that in the CT and BCG1 groups, and the GLUT-2 and LAT-1 relative mRNA levels were greater in the BCG2 group than the CT group (P < 0.05). Dietary B. licheniformis BCG resulted in significantly higher ileal occludin, and lower IL-8 and TLR-4 mRNA levels than observed in the CT group (P < 0.05). B. licheniformis BCG supplementation significantly decreased bacterial community richness and diversity in the ileum (P < 0.05). Dietary B. licheniformis BCG shaped the ileac microbiota by increasing the prevalence of f_Sphingomonadaceae, Sphingomonas, and Limosilactobacillus, and contributed to nutrient digestion and absorption; moreover, it enhanced the intestinal barrier by increasing the prevalence of f_Lactobacillaceae, Lactobacillus, and Limosilactobacillus. Dietary B. licheniformis BCG decreased microbial community diversity by diminishing Desulfovibrio, Alistipes, Campylobacter, Vibrio, Streptococcus, and Escherichia coli-Shigella levels, and down-regulating inflammatory associated molecule expression. Therefore, dietary B. licheniformis BCG contributed to digestion and absorption of nutrients, enhanced the intestinal physical barrier, and decreased intestinal inflammation in broilers by decreasing microbial diversity and optimizing the microbiota structure.

Butyrate Mitigates Lipopolysaccharide-Induced Intestinal Morphological Changes in Weanling Piglets by Regulating the Microbiota and Energy Metabolism, and Alleviating Inflammation and Apoptosis.

Butyrate provides energy for colonocytes and is a functional metabolite that mitigates weanling piglet stress. However, its effects and mechanisms remain largely unknown. We established a lipopolysaccharide (LPS)-induced inflammatory stress piglet model to examine how butyrate mechanisms impacted piglet intestinal histology, microbiota, and inflammation. We randomly assigned 18 crossbred male piglets to three treatment groups: CON, LPS, and BT-LPS. Coated butyrate was supplemented in the BT-LPS feed for 21 days. On days 19 and 21, piglets in LPS and BT-LPS groups were challenged with LPS at 100 µg/kg body weight. Dietary butyrate improved LPS-injured intestinal histology by significantly increasing jejunal and ileal villus height, villus height to crypt depth ratios, and decreasing histological scores. LPS challenge activated hypoxia-inducible factor 1α and nuclear factor-κB, and enhanced interleukins (IL-1ß, IL-6, IL-12), tumor necrosis factor-α, and also downstream inducible nitric oxide synthase and cyclooxygenase 2, but decreased anti-inflammatory cytokines (IL-10, IL-13). Most molecule levels were significantly reversed by butyrate administration. When compared with the CON or LPS groups, the BT-LPS group had a higher relative abundance of jejunal Firmicutes, Bacteroidetes, Clostridiaceae, Lactobacillus, and Prevotella but a lower abundance of Proteobacteria, Enterobacteriaceae, and Escherichia-Shigella. Phylogenetic investigation of communities by reconstruction of unobserved states and correlation analyses suggested these bacteria contributed to butyrate-alleviating jejunal inflammation and infectious diseases. Butyrate-based diets significantly reduced apoptosis via mitochondrial pathways by downregulating apoptotic caspase 3 mRNA levels. Diets also altered enterocyte metabolism in the jejunum by upregulating peroxisome-proliferator-activated receptor α expression but downregulating carnitine palmitoyltransferase 1 level when compared with CON or LPS groups. Butyrate supplementation improved immunity homeostasis, generated beneficial shifts in microbial communities, improved enterocyte energy metabolism, and prevented apoptosis to protect intestinal histology from LPS-induced injury.

Metabolomic Characteristics of Liver and Cecum Contents in High-Fat-Diet-Induced Obese Mice Intervened with Lactobacillus plantarum FRT10.

Obesity has become a major social problem related to health and quality of life. Our previous work demonstrated that Lactobacillus plantarum FRT10 alleviated obesity in high-fat diet (HFD)-fed mice by alleviating gut dysbiosis. However, the underlying functions of FRT10 in regulating liver and cecum contents metabolism remain unknown. Liver and cecum contents metabonomics combined with pathway analysis based on ultraperformance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UHPLC-Q-TOF/MS) were performed to evaluate the alterations of metabolic profiles between obese control mice and obese mice in FRT10-treated groups. The orthogonal partial least squares discriminant analysis (OPLS-DA) score plots showed that there were significant differences in cecum contents and liver markers between experimental groups. In total, 26 potential biomarkers were identified in the liver and 15 in cecum contents that could explain the effect of FRT10 addition in HFD-fed mice. In addition, gut-liver axis analysis indicated that there was a strong correlation between cecum contents metabolites and hepatic metabolites. The mechanism of FRT10 against obesity might be related to the alterations in glycerophospholipid metabolism, primary bile acid biosynthesis, amino metabolism, and purine and pyrimidine metabolism. Studies on these metabolites could help us better understand the role of FRT10 in obesity induced by HFD.

Fermented Soy and Fish Protein Dietary Sources Shape Ileal and Colonic Microbiota, Improving Nutrient Digestibility and Host Health in a Piglet Model.

Suitable protein sources are essential requirements for piglet growth and health. Typically, intestinal microbiota co-develops with the host and impact its physiology, which make it more plastic to dietary protein sources at early stages. However, the effects of fermented soybean meal (FSB) and fish meal (FM) on foregut and hindgut microbiota, and their relationship with nutrient digestion and host health remain unclear. In this study, we identified interactions between ileac and colonic microbiota which were reshaped by FSB and FM, and assessed host digestibility and host health in a piglet model. Eighteen weaned piglets (mean weight = 8.58 ± 0.44 kg) were divided into three dietary treatments, with six replicates/treatment. The level of dietary protein was 16%, with FSB, FM, and a mixture of fermented soybean meal and fish meal (MFSM) applied as protein sources. During days 1-14 and 1-28, diets containing MFSM generated higher piglet body weight and average daily gain, but lower feed to weight gain ratios when compared with the FM diet (P < 0.05). Piglets in MFSM and FM groups had lower apparent total tract digestibility (ATTD) of crude protein (CP) compared with the FSB group (P < 0.05). Serum immunoglobulins (IgM and IgG) in MFSM and FM groups were significantly higher on day 28, but serum cytokines (interleukin-6 and tumor necrosis factor-α) were significantly lower than the FSB group on days 14 and 28 (P < 0.05). When compared with FSB and FM groups, dietary MFSM significantly increased colonic acetic acid and butyric acid levels (P < 0.05). Compared with the FM and MFSM groups, the FSB diet increased the relative abundance of ileac Lactobacillus and f_Lactobacillaceae, which were significant positively correlated with CP ATTD (P < 0.05). Compared with the FSB group, the relative abundance of f_Peptostreptococcaceae and Romboutsia in MFSM or FM groups were increased and were significant positively correlated with total carbohydrate (TC) ATTD (P < 0.05). Piglets fed FSB had higher α-diversity in colonic microbiota when compared with other groups (P < 0.05). The relative abundance of colonic unidentified_Clostridiales and Romboutsia in MFSM and FSB groups were significantly higher than in the FM group (P < 0.05). Dietary MFSM or FM increased the relative abundance of colonic Streptococcaceae and Streptococcus, but decreased the relative abundance of Christensenellaceae when compared with the FSB group (P < 0.05). These bacteria showed a significantly positive correlation with serum cytokine and immunoglobulin levels (P < 0.05). Therefore, dietary FSB improved CP digestibility by increasing the relative abundance of ileac f_Lactobacillaceae and Lactobacillus, while dietary MFSM benefited TC digestibility by increasing f_Peptostreptococcaceae and Romboutsia. Dietary MFSM and FM enhanced immunoglobulin secretion by increasing colonic f_Streptococcaceae and Streptococcus prevalence, while dietary FSB promoted cytokine production by increasing microbiota diversity and Romboutsia and Christensenellaceae. Our data provide a theoretical dietary basis for young animals using plant and animal protein sources.

Multi-Omics Profiling Reveals Se Deficiency-Induced Redox Imbalance, Metabolic Reprogramming, and Inflammation in Pig Muscle.

BACKGROUND: Nutritional muscle dystrophy is associated with selenium (Se) deficiency; however, the underlying mechanism remains unclear. OBJECTIVES: This study aimed to understand the crosstalk among redox status, energy metabolism, and inflammation in nutritional muscle dystrophy induced by dietary Se deficiency. METHODS: Eighteen castrated male pigs (Yorkshire, 45 d old) were fed Se-deficient (Se-D; 0.007 mg Se/kg) or Se-adequate (Se-A; in the form of selenomethionine, 0.3 mg Se/kg) diets for 16 wk. The muscle Se concentrations; antioxidant capacity; and gene expression, transcriptome, global proteome, metabolome, and lipidome profiles were analyzed. The transcriptome, metabolome, and proteome profiles were analyzed with biostatistics, bioinformatics, and pathway enrichment analysis; other data were analyzed with Student's 2-sided t tests. RESULTS: The muscle Se content in the Se-D group was 96% lower than that in the Se-A group (P < 0.05). The activity of glutathione peroxidase (GPX) and thioredoxin reductase (TXNRD) in the Se-D group was 42%-69% lower than that in the Se-A group (P < 0.05). The mRNA levels of 10 selenoprotein genes were 25%-84% lower than those in the Se-A group (P < 0.05). Multi-omics analyses indicated that the levels of 1378 transcripts, 83 proteins, 22 metabolites, and 55 lipid molecules were significantly altered in response to Se deficiency. Se deficiency-induced redox imbalance led to muscle central carbon and lipid metabolism reprogramming, which enhanced the glycolysis pathway and decreased phospholipid synthesis. Inflammation and apoptosis were observed in response to Se deficiency-induced muscle oxidative stress, which may have been associated with extracellular matrix (ECM) remodeling, suppressed focal adhesion and phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling, and activation of the NF-κB signaling pathway. CONCLUSIONS: These results contributed to understanding the crosstalk among redox, energy metabolism, and inflammation in Se deficiency-induced muscle dystrophy in pigs, and may provide intervention targets for muscle disease treatment.

Selenium Deficiency Induces Pathological Cardiac Lipid Metabolic Remodeling and Inflammation.

SCOPE: Selenium (Se) disequilibrium is closely involved in many cardiac diseases, although its in vivo mechanism remains uncertain. Therefore, a pig model is created in order to generate a comprehensive picture of cardiac response to dietary Se deficiency. METHODS AND RESULTS: A total of 24 pigs are divided into two equal groups, which were fed a diet with either 0.007 mg kg-1 Se or 0.3 mg kg-1 Se for 16 weeks. Se deficiency causes cardiac oxidative stress by blocking glutathione and thioredoxin systems and increases thioredoxin domain-containing protein S-nitrosylation. Energy production is disordered, as free fatty acids are overloaded, the tricarboxylic acid cycle is strengthened, and three respiratory chain proteins enhance S-nitrosylation. Excess free fatty acids lead to increased synthesis of diacylglycerol, phosphatidylcholine, and phosphatidylethanolamine, where the latter two are vulnerable to oxidation and causes an increase in malondialdehyde. Moreover, increased palmitic acid enhances de novo ceramide synthesis and accumulation. Additionally, Se deficiency initiates inflammation via cytosolic DNA-sensing pathways, which activates downstream interferon regulatory factor 7 and nuclear factor kappa B. CONCLUSIONS: The present study identifies a lipid metabolic vulnerability and inflammation initiation pathway via Se deficiency, which may provide targets for human redox imbalance-induced cardiac disease treatment.

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.

Butyrate Mitigates Weanling Piglets From Lipopolysaccharide-Induced Colitis by Regulating Microbiota and Energy Metabolism of the Gut-Liver Axis.

Inflammatory bowel disorder is accompanied by the destruction of immunity homeostasis, gut microbiota perturbation, and chronic inflammatory liver diseases. Butyrate is known as a primary energy source for colonocytes and functional substances for mitigating pathological features of colitis. However, it is still unclear whether butyrate alleviates colitis progression by regulation of microbiota and metabolism in the gut-liver axis. In the present study, we aimed to determine the role of microbiota and metabolism of the gut-liver axis in ameliorating lipopolysaccharide (LPS)-induced colitis in piglets using protected butyrate administration. Eighteen crossbred male piglets were weaned at 30 days old and were randomly allocated to three treatments, with CON (basal diet), LPS (basal diet + LPS), and BT-LPS (basal diet + 3.0 g/kg protected butyrate + LPS). On days 19 and 21, piglets in the LPS and BT-LPS groups were intraperitoneally challenged with LPS at 100 µg/kg body weight. Butyrate administration significantly decreased LPS-induced rise in the clinical score of piglets and colonic histological scores and reduced the susceptibility to LPS-induced severe inflammatory response by decreasing proinflammatory (IL-1ß, IL-6, IL-8, and TNF-α) cytokines. Butyrate supplementation accelerated the prevalence of Faecalibacterium and Lactobacillus by enhancing the tricarboxylic acid (TCA) cycle of colonocytes. Dietary supplementation with protected butyrate significantly targeted increased concentrations of butyric acid in the colon and portal venous circulation, and enhanced the TCA cycle in the gut-liver axis by mobilizing amino acid and vitamin B group as a coenzyme. Meanwhile, during this progress, LPS increased fatty acid synthesis that was reversed by butyrate treatment, which was reflected by decreased acylcarnitines. Butyrate-reshaped colonic microbial community and metabolism in the gut-liver axis contributed to morphology integrity and immunity homeostasis by promoting anti-inflammatory (IL-10 and TGF-ß) cytokines and suppressing inflammatory mediator hypoxia-inducible factor 1α and its downstream response elements cyclooxygenase 2 and inducible nitric oxide synthase. These results identified the pivotal role of colonic microbiota and metabolism in the gut-liver axis for alleviating inflammatory progression and possible therapeutic targets.

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.

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.

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.

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.

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.

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.

[Ultrasound and MRI features of malignant fibrous histiocytoma of soft tissue].

OBJECTIVE: To investigate ultrasound and MRI features of malignant fibrous histiocytoma (MFH) of soft tissue. METHODS: Ultrasound, MRI images and pathological data of 12 patients with malignant fibrous histiocytoma in soft tissue confirmed by operation and pathology were analyzed from January 2012 to August 2018, inlcuding 7 males and 5 females, aged from 36 to 69 years old with an average age of 53 years old; the courses of disease ranged from 4 to 49 months with an average of 28 months. Clinical manifestations were soft tissue masses and pain in the affected limbs. Ultrasound, MRI and contrast-enhanced examination were performed before operation. The lesions, morphology, echo/signal characteristics, color flow signals and enhancement features were observed and compared with pathology. RESULTS: In 12 patients with MFH, 9 patients were primary lesions and 3 patients were recurrent lesions after operation. There were 7 cases of bilateral thighs, 2 cases of calves, 1 case of upper arm, 1 case of buttocks and 1 case of posterior peritoneum. The size ranged from 5.1 to 17.1 cm with an average of 8.7 cm. Ultrasound feature showed lobulated or agglomerate, and focused on low echo; 5 cases had capsule and with clear border; 7 cases were unclear boundary with surrounding tissues; and 6 cases with irregular echo-free. The blood flow signals were around the CDFI, and the internal blood flow signals were different. MRI feature showed lobulated, agglomerate or irregular shape, T1WI showed slightly lower signal or equal signal, T2WI showed high signal and DWI signal increased. Six patients manifested mixed signal inside, 7 patients manifested low signal separation inside, 5 patients with false envelope, and 9 patients manifested infiltration and growth with peripheral edema. T1WI showed uneven strengthening after enhancement. Immunohistochemical expression of Vim, CD68 were positive. CONCLUSIONS: The age, location and imaging features of soft tissue MFH are characteristic. The diagnosis of MFH should be considered when irregular mass occurred in soft tissues of limbs at middle-aged and old people. Echo and signal are homogeneous or mixed. Separation, necrosis and cystic degeneration could be seen in the mass. When the blood flow signals are abundant and solid components are obviously enhanced, the diagnosis of MFH should be considered.

The use of hair as a long-term indicator of low-dose ß2 agonist treatments in cattle: Implications for growth-promoting purposes monitoring.

The objective of this study was to assess the feasibility of using hair as a long-term indicator of cocktail (low-dose ß2 agonists) treatments in cattle. Six male Simmental cattle were treated with a mixture of low-dose clenbuterol, ractopamine, and salbutamol at dosages of 5.3, 223.3, and 50.0 µg/kg, respectively. The trial lasted for 112 days and included 28 days of treatment and 84 days of withdrawal. Plasma and urine samples taken during the treatment period contained the highest residues, with maximum concentrations of clenbuterol, ractopamine, and salbutamol in plasma of 1.49 ng/mL (Day 21), 43.78 (Day 14) ng/mL, and 8.07 ng/mL (Day 7), respectively, and in urine of 62.40 ng/mL (Day 28), 3995.77 ng/mL (Day 28), and 503.72 ng/mL (Day 1), respectively. On day 42 of withdrawal, the residues of all three ß2 agonists in plasma were below the limit of quantification (LOQ; 0.3 ng/mL for clenbuterol, and 0.5 ng/mL for ractopamine and salbutamol), and in urine samples were below or near the LOQ (the highest being ractopamine at 1.10 ng/mL). The highest concentrations of clenbuterol, ractopamine, and salbutamol in hair were 88.36, 1351.92, and 100.58 ng/g, respectively, on day 14 of withdrawal; and the residues were long-lasting, with 7.64, 28.55, and 8.77 ng/g, respectively, on day 84 of withdrawal. The results of this study demonstrate that hair could be utilized as a long-term indicator of the use of a combination of low-dose ß2 agonists in cattle, which could have implications for growth-promoting purposes monitoring.

miR-224 Affects Mammary Epithelial Cell Apoptosis and Triglyceride Production by Downregulating ACADM and ALDH2 Genes.

MicroRNAs (miRNAs) are small noncoding RNA molecules that involve in various biological functions by regulating the expressions of target genes. In recent years, many researchers have demonstrated that miR-224 played an important role in regulating lipid metabolism. Therefore, in this study, the target genes of miR-224 were verified and the regulatory role of miR-224 was confirmed in lipid metabolism. In this study, bioinformatics methods were used for primarily predicting the target gene of miR-224 and dual-luciferase reporter system was used for further verify the relationship between miR-224 and its target gene. Then, the miR-224 mimics, miR-224 inhibitor, and miRNA-ShNC were transfected into mammary epithelial cells (MECs), respectively, and the expression of miR-224 and its target genes was detected by quantitative real-time polymerase chain reaction and Western blot. Furthermore, the triglyceride production and cell apoptosis were detected by triglyceride mensuration reagent kit using flow cytometry. The results showed that ACADM and ALDH2 were predicted to be the target genes of miR-224, primarily by bioinformatics analysis. We founded that miR-224 could recognize with ACADM-3'UTR and ALDH2-3'UTR, indicating that the target sites existed in 3'UTR of ACADM and ALDH2. And then, the expressions of miR-224 had negative trend with the levels of ACADM and ALDH2, suggesting that miR-224 could downregulate the expressions of ACADM and ALDH2. Finally, the triglyceride production decreased and apoptosis rate increased after the overexpression of miR-224 in MECs. The above results indicated that miR-224 regulating target genes in lipid metabolism might be used as a new pathway for better breeding.