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IntroductionThe Tibetan Plateau is characterized by low temperature and hypoxia. N-carbamylglutamic acid (NCG) can increase blood oxygen saturation, and have the potential to be used to prevent the high-altitude hypoxia stress state of cows. However, its beneficial effect on the rumen microbiota of Holstein dairy cows remains unclear.MethodsHence, the experiments 12 multiparous (parity ranged from 2 to 7) Holstein dairy cows (413.0 ± 42 kg) were randomly assigned to 2 treatments with 6 replicates in each treatment: basal diet (CON, control group) and basal diet plus 20 g/d/cow of NCG (NCG, experiment group), respectively. To study the effects of dietary NCG supplementation on rumen microbiota of Holstein dairy cows in Tibet. The experiment lasted for 45 days, with 15 days of pre-feeding and 30 days of formal trail period.ResultsThe results showed that ruminal NH3-N concentration in NCG group was lower (p p p Bacteroidetes, Firmicutes, and Patescibacteria as the most abundant phyla. The KEGG pathway analysis showed that the potential function of ruminal bacteria was mainly enriched in metabolism (carbohydrates, amino acids, lipids, energy, and nucleotides) and genetic information processing (replication, repair, and translation).ConclusionIn conclusion, NCG can improve rumen nitrogen utilization, total VFA and acetic acid production, and increase rumen microbial diversity, all of which could make the introduced Holstein dairy cows to better adapt to the harsh environment in Tibet and improve their production performance.

In order to study the regulation of Fenugreek seed extract (FSE) on the immunity of broilers, and explore the appropriate amount of FSE in broilers' production, 1-day-old yellow feather broilers with a total of 420 birds were randomly allocated into seven treatments. Each treatment had six replicates, with 10 birds per replicate. The two control groups were the basic fodder group fed with basal diet and the bacitracin zinc group added 30 mg/kg bacitracin zinc to the basal diet. Experimental groups included five levels of FSE (50, 100, 200, 400, and 800 mg/kg FSE to the basal diet, respectively). The pre-test period was 7 days and the formal test lasted for 56 days. The results showed that the average daily gain (ADG) of 50 and 800 mg/kg FSE groups was significantly increased (P < 0.01), and the feed to gain ratio (F/G) of FSE groups was significantly decreased (P < 0.01) compared with the basic fodder and the bacitracin zinc groups. Compared with the basic fodder group, the serum total cholesterol (TC) content in the FSE groups was significantly decreased (P < 0.05), the serum low density lipoprotein cholesterol (LDL-C) content of 50, 100, and 800 mg/kg FSE groups was significantly lower than that of the basic fodder group (P < 0.05). Compared with the basic fodder and bacitracin zinc groups, the serum immunoglobulins (IgG, IgM, IgA) content of 100 and 200 mg/kg FSE groups were significantly increased (P < 0.05). Compared with the bacitracin zinc group, the serum interleukins (IL-1, IL-10) content of 400 mg/kg FSE group were significantly increased (P ≤ 0.05), and the serum interferon-γ (IFN-γ) content of 100 and 200 mg/kg FSE groups was significantly increased (P < 0.05). Compared with the basic fodder group, the lower doses (0–400 mg/kg) of FSE had no significant effect on the mRNA expression of toll-like receptors 4/ myeloid differentiation factor 88/ nuclear factor-κB (TLR4/MyD88/NF-κB) signaling pathways (P > 0.05). The 800 mg/kg FSE treatment group significantly increased the expression levels of nuclear factor-κB (NF-κB) mRNA in the spleen of broilers (P < 0.05). The zinc bacitracin group significantly increased the expression levels of myeloid differentiation factor 88 (MyD88) and nuclear factor-κB (NF-κB) mRNA (P ≤ 0.05). The results showed that FSE could promote the secretion of immunoglobulins, regulate the body's cytokines, and have a positive effect on immunity in broilers. Furthermore, the recommended supplement of FSE is 100 mg/kg in the broiler diet.

Resveratrol (RSV) is a natural polyphenolic compound with potent antioxidant and anti-inflammatory properties. This study aimed to investigate the protective effects of RSV supplementation on the inflammatory responses of broilers during heat stress. A total of 432 28-d-old white-feathered broilers (817 crossbred chicken) with an average weight of 549 ± 4 g were randomly allotted to 4 equal groups (Half male and half female, 6 replicates/group, 18 chickens/replicate), including normal temperature (NT) group (24 ± 2°C for 24 h/d, basal diet), NT+RSV group (24 ± 2°C for 24 h/d, basal diet + RSV), heat stress (HT) group (37 ± 2°C for 8 h/d, basal diet), and HT+RSV group (37 ± 2°C for 8 h/d, basal diet + RSV). Serum samples were collected on d 7 and 14 of heat stress, and thymus, spleen, jejunum, and bursa of Fabricius samples were collected and analyzed on d14. RSV treatment decreased the feed conversion ratio, partially reversed the negative alternations in body weight, average daily gain, and average daily feed intake caused by heat stress. RSV treatment also decreased the elevated levels of corticosterone on d 14, adrenocorticotropic hormone, and triiodothyronine in serum on d 7 caused by heat stress, and significantly increased the villus height to crypt depth ratio in the jejunum on d 14. Dietary RSV also reduced heat stress-induced splenic pro-inflammatory cytokine concentrations. TUNEL assay showed that RSV significantly reduced heat stress-induced the number of apoptotic cells. Remarkably, RSV down-regulated some splenic related genes for apoptosis genes, including BCL-2, Apaf-1, and MDM2 mRNA levels induced by heat stress. According to GO and KEGG enrichment analyses, the differential genes between HT and HT + RSV groups were mainly associated with immune system process, hematopoietic or lymphoid organ development, and toll-like receptor signaling pathway. The relative mRNA expression of NF-κB, heat shock protein 70 (HSP70), and p38 MAPK were markedly decreased by the combination of RSV and heat stress. These findings showed that RSV might reduce the splenic inflammatory response in heat-stressed white-feather broilers by inhibiting heat stress-induced activation of NF-B, MAPK, and HSP70, as well as inhibiting the activation of mitochondrial apoptotic pathways.

The current study aimed to investigate the solid-state fermentation process of Quercus liaotungensis (QL) by Bacillus subtilis (BS). The parameters included the inoculation amount, the soybean meal addition amount, the fermentation temperature and the ratio of material to water. The optimal process was determined based on the nutritional value, tannin content and DPPH clearance of QL after fermentation. The results showed that: (1) The parameters of the optimal process included inoculating 106 BS per gram of QL, then adding 10% soybean meal, the ratio of material to the water of 100:80, and temperature at 33 °C for 72 h. (2) In the optimum fermentation conditions, the crude fiber content, and the ether extract content of QL decreased by 66.94% and 66.96%, respectively (p < 0.05). Moreover, the crude protein content and the ash content increased by 65.81% and 4.63%, respectively, after fermentation (p < 0.05). Additionally, the tannin content decreased by 62.77% (p < 0.05), and the DPPH scavenging rate decreased by 45.45% (p < 0.05) after fermentation, respectively. In summary, the QL significantly improved the nutritional value after the solid-state fermentation with BS.

This study was conducted to investigate how chito-oligosaccharides (COSs) affect the growth performance and immune stress response and to further explain their mechanisms. A total of 32 boars that were 28 days old and three-way weaned were randomly allotted to four equal groups [CON (basal diet) group, enterotoxigenic

This study was conducted to evaluate the effect of mulberry leaves as an alternative source of protein on growth performance, carcass traits and meat quality in finishing pigs. A total of 180 Xiangcun Black pigs were randomly assigned to five treatment groups with six pens of six pigs per pen. The pigs were provided with a basal diet or a diet contained 3%, 6%, 9% or 12% of mulberry leaf powder during a 50-day experiment period. The results showed that dietary mulberry leaf powder had no negative effect on growth performance in Xiangcun Black pigs, except in the 12% mulberry group, where final body weight and average daily gain decreased (p < .05) and feed to gain ratio of the pigs increased (p < .05). Dietary mulberry inclusion decreased (quadratic, p < .05) the back fat thickness, fibre mean cross-sectional area (CSA) in the longissimus dorsi (LD) muscle and mRNA expression levels of myosin heavy chain (MyHC) IIb in LD and biceps femoris (BF) muscles, while increased (linear or quadratic, p < .05) the plasma concentration of albumin, levels of crude protein (CP), inosine monophosphate (IMP) and several amino acids in muscle tissues. When compared with the other groups, the 9% mulberry diet increased (p < .05) loin-eye area and contents of CP and IMP in muscles, while decreased (p < .05) plasma activity of cholinesterase and concentrations of uric acid and urea. The 6% mulberry diet had the lowest fibre mean CSA and shear force and increased total fibre number of the LD muscle, when compared with the other groups. These results suggest that including mulberry in the diet at

Background: Eucommia ulmoides flavones (EUF), have been demonstrated to attenuate the inflammation and oxidative stress of piglets. This study was designed to test whether EUF could be used as an antibiotic growth promoter alternative to support growth performance and maintain intestinal health in weanling-piglets. Weaned-piglets (n = 480) were assigned into 3 groups and fed with a low-protein basal diet (NC), or supplementation with antibiotics (75 mg/kg quinocetone, 20 mg/kg virginomycin and 50 mg/kg aureomycin, PC) or 0.01% EUF (EUF). Blood, intestinal contents and intestine were collected on d-15 and d-35 after weaning, respectively. Results: The results showed that the body weight on d-35, average daily gain and gain : feed ratio from d 15 to d 35 and d 0 to d 35 in piglets of PC and EUF treatments were higher than (P < 0.05) these in NC treatment, whereas the initial body weight among 3 treatments were consistent. PC and EUF treatments decreased (P < 0.05) the diarrhea index of weanling piglets. Dietary supplementation of EUF significantly enhanced (P < 0.05) the serum concentrations of total protein, alanine transaminase and the immunoglobulin G on d-15 of piglets compared to piglets in NC treatment. EUF supplementation increased (P < 0.05) the jejunal villus height : crypt depth ratio, jejunal and ileal villus height and the population of ileal lactic acid bacteria on d-15 but remarkable decreased (P < 0.05) the population of ileal coliform bacteria on d-15 and d-35. Conclusion: These findings indicated the EUF, as the potential alternative to in-feed antibiotic growth promoter, could improve growth performance and intestinal morphology, decreasing colonization of coliform bacteria and diarrhea index in weanling piglets.

Background Eucommia ulmoides flavones (EUF), have been demonstrated to attenuate the inflammation and oxidative stress of piglets. This study was designed to test whether EUF could be as an antibiotic alternative to support growth performance and maintain intestinal health in weanling-piglets. Weaned-piglets (n = 480) were assigned into 3 groups and fed with a low-protein basal diet (NC), or supplementation with antibiotics (PC) or 0.1‰ EUF (EUF). Blood, intestinal contents and intestine were collected on d-15 and d-35, respectively. Results The results showed that the body weight on d-35, average daily gain and gain : feed from d 15 to d 35 and d 0 to d 35 in piglets of PC and EUF treatments were higher than (p p p p p coliform bacteria and diarrhea index in weanling piglets.

The synthetic dipeptides alanyl-glutamine (Ala-Gln) and glycyl-glutamine (Gly-Gln) are used as Gln substitution to provide energy source in the gastrointestinal tract due to their high solubility and stability. This study aimed to investigate the effects of Gln, Ala-Gln and Gly-Gln on mitochondrial respiration and protein turnover of enterocytes. Intestinal porcine epithelial cells (IPEC-J2) were cultured for 2 days in Dulbecco's modified Eagle's-F12 Ham medium (DMEM-F12) containing 2.5 mM Gln, Ala-Gln or Gly-Gln. Results from 5-ethynyl-2'-deoxyuridine incorporation and flow cytometry analysis indicated that there were no differences in proliferation between free Gln and Ala-Gln-treated cells, whereas Gly-Gln treatment inhibited the cell growth compared with Gln treatment. Significantly lower mRNA expressions of Sp1 and PepT1 were also observed in Gly-Gln-treated cells than that of Ala-Gln treatment. Ala-Gln treatment increased the basal respiration and ATP production, compared with free Gln and Gly-Gln treatments. There were no differences in protein turnover between free Gln and Ala-Gln-treated cells, but Gly-Gln treatment reduced protein synthesis and increased protein degradation. Ala-Gln treatment stimulated mTOR activation whereas Gly-Gln decreased mTOR phosphorylation and increased the UB protein expression compared with free Gln treatment. These results indicate that Ala-Gln has the very similar functional profile to free Gln in porcine enterocytes in vitro and can be substituted Gln as energy and protein sources in the gastrointestinal tract.

Simple Summary N-carbamylglutamate (NCG) has been demonstrated to promote the synthesis of endogenous arginine and improve reproductive performance. In the present study, we found that dietary NCG supplementation improved meat quality of a Chinese fat-type pig by increasing muscle tenderness and Phe concentration, and optimizing fatty acid profiles in different tissues. These results provided scientific evidence for the application of NCG as a feed additive in finishing pigs. Abstract The present study evaluated the effects of dietary N-carbamylglutamate (NCG) on carcass traits, meat quality, and fatty acid profiles in the longissimus dorsi muscle and adipose tissues of Chinese Ningxiang pigs. A total of 36 castrated female pigs with a similar initial weight (43.21 ± 0.57 kg) were randomly assigned to two treatments (with six pens per treatment and three pigs per pen) and fed either a basal diet or a basal diet supplemented with 0.08% NCG for 56 days. Results showed that dietary NCG reduced shear force (p = 0.004) and increased drip loss (p = 0.044) in longissimus dorsi muscle of Ningxiang pigs. Moreover, increased levels of oleic acid (C18:1n9c) (p = 0.009), paullinic acid (C20:1) (p = 0.004), and α-linolenic acid (C18:3n3) (p < 0.001), while significant reduction in the proportions of arachidonic acid (C20:4n6) (p < 0.001) and polyunsaturated fatty acid (PUFA) (p = 0.017) were observed in the longissimus dorsi muscle of pigs fed NCG when compared with those fed the control diet. As for adipose tissues, the C20:1 (p = 0.045) proportion in dorsal subcutaneous adipose (DSA), as well as the stearic acid (C18:0) (p = 0.018) level in perirenal adipose (PA) were decreased when pigs were fed the NCG diet compared with those of the control diet. In contrast, the margaric acid (C17:0) (p = 0.043) proportion in PA were increased. Moreover, the NCG diet produced PA with a greater proportion of total PUFAs (p = 0.001) (particularly linoleic acid (C18:2n6c) (p = 0.001)) compared with those produced by the control diet. These findings suggest that dietary NCG has beneficial effects by decreasing the shear force and improving the healthfulness of fatty acid profiles, providing a novel strategy for enhancing meat quality of pigs.

The present study was conducted to evaluate the effects of glucose, soya oil or glutamine on jejunal morphology, protein metabolism and protein expression of the mammalian target of rapamycin complex 1 (mTORC1) signalling pathway in jejunal villus or crypt compartment of piglets. Forty-two 21 d-weaned piglets were randomly allotted to one of the three isoenergetic diets formulated with glucose, soya oil or glutamine for 28 d. On day 14 or 28, the proteins in crypt enterocytes were analysed with isobaric tags for relative and absolute quantification and proteins involved in mTORC1 signalling pathway in villus or crypt compartment cells were determined by Western blotting. Our results showed no significant differences (P > 0·05) in jejunal morphology among the three treatments on day 14 or 28. The differentially expressed proteins mainly took part in a few network pathways, including antimicrobial or inflammatory response, cell death and survival, digestive system development and function and carbohydrate metabolism. On day 14 or 28, there were higher protein expression of eukaryotic initiation factor-4E binding protein-1 in jejunal crypt compartment of piglets supplemented with glucose or glutamine compared with soya oil. On day 28, higher protein expression of phosphor-mTOR in crypt compartment was observed in piglets supplemented with glucose compared with the soya oil. In conclusion, the isoenergetic glucose, soya oil or glutamine did not affect the jejunal morphology of piglets; however, they had different effects on the protein metabolism in crypt compartment. Compared with soya oil, glucose or glutamine may be better energy supplies for enterocytes in jejunal crypt compartment.

Eucommia ulmoides flavones (EUF) have been demonstrated to alleviate oxidative stress and intestinal damage in piglets, but their effect target is still poorly understood. NF-E2-related factor 2 (Nrf2) pathway plays a very important role in the defense mechanism. This study was designed to investigate the regulation of EUF on the Nrf2 pathway and inhibition of Nrf2 on oxidative stress in the intestine of piglets. An in vivo study was conducted in weaned piglets treated with basal diet, basal diet+diquat, and 100 mg/kg EUF diet+diquat for 14 d to determine Nrf2 and Keap1 protein expressions, as well as downstream antioxidant gene mRNA expression. An in vitro study was performed in a porcine jejunal epithelial cell line to investigate the effect of inhibiting Nrf2 on cell growth and intracellular oxidative stress parameters. The results showed that the supplementation of EUF decreased the oxidized glutathione (GSSG) concentration and the ratio of GSSG to glutathione (GSH) but increased the protein expressions of nuclear Nrf2 and Kelch-like ECH-associated protein 1 (Keap1) as well as mRNA expression of heme oxygenase 1 (HO-1), NAD(P)H:quinone oxidoreductase 1 (NQO-1), and glutamate cysteine ligase catalytic subunit (GCLC) in the small intestinal mucosa of diquat-challenged piglets. When Nrf2 was inhibited by using ML385, cell viability, cellular antioxidant activities, expressions of nuclear Nrf2 and Keap1 protein, and downstream antioxidant enzyme (HO-1, NQO-1, and GCLC) mRNA were decreased in paraquat-treated enterocytes. These results showed that the Nrf2 signaling pathway played an important role in EUF-regulating oxidative stress in the intestine of piglets.

Resveratrol (3,4′,5-trihy- droxystilbene), a natural phytoalexin polyphenol, exhibits anti-oxidant, anti-inflammatory, and anti-carcinogenic properties. This phytoalexin is well-absorbed and rapidly and extensively metabolized in the body. Inflammation is an adaptive response, which could be triggered by various danger signals, such as invasion by microorganisms or tissue injury. In this review, the anti-inflammatory activity and the mechanism of resveratrol modulates the inflammatory response are examined. Multiple experimental studies that illustrate regulatory mechanisms and the immunomodulatory function of resveratrol both in vivo and in vitro. The data acquired from those studies are discussed.

The present study evaluated the effects of dietary N-carbamylglutamate (NCG) on carcass traits, meat quality, and fatty acid profiles in the longissimus dorsi muscle and adipose tissues of Chinese Ningxiang pigs. A total of 36 castrated female pigs with a similar initial weight (43.21 &plusmn, 0.57 kg) were randomly assigned to two treatments (with six pens per treatment and three pigs per pen) and fed either a basal diet or a basal diet supplemented with 0.08% NCG for 56 days. Results showed that dietary NCG reduced shear force (p = 0.004) and increased drip loss (p = 0.044) in longissimus dorsi muscle of Ningxiang pigs. Moreover, increased levels of oleic acid (C18:1n9c) (p = 0.009), paullinic acid (C20:1) (p = 0.004), and &alpha, linolenic acid (C18:3n3) (p &lt, 0.001), while significant reduction in the proportions of arachidonic acid (C20:4n6) (p &lt, 0.001) and polyunsaturated fatty acid (PUFA) (p = 0.017) were observed in the longissimus dorsi muscle of pigs fed NCG when compared with those fed the control diet. As for adipose tissues, the C20:1 (p = 0.045) proportion in dorsal subcutaneous adipose (DSA), as well as the stearic acid (C18:0) (p = 0.018) level in perirenal adipose (PA) were decreased when pigs were fed the NCG diet compared with those of the control diet. In contrast, the margaric acid (C17:0) (p = 0.043) proportion in PA were increased. Moreover, the NCG diet produced PA with a greater proportion of total PUFAs (p = 0.001) (particularly linoleic acid (C18:2n6c) (p = 0.001)) compared with those produced by the control diet. These findings suggest that dietary NCG has beneficial effects by decreasing the shear force and improving the healthfulness of fatty acid profiles, providing a novel strategy for enhancing meat quality of pigs.

Context Heat stress is one of the problems commonly found in broiler industry in tropic and subtropic regions that results in impairment of intestinal integrity, leading to inflammation and poor performance. Aims This study aimed to investigate the effect of dietary resveratrol supplementation on growth performance, intestinal morphology and barrier integrity, and inflammation response of yellow-feather broilers under heat stress. Methods In total, 288 birds (28-day-old) were randomly allotted to three treatment groups, with six replicates. A thermo-neutral group (24 ± 2°C) received a basal diet and two heat-stressed groups (37 ± 2°C for 8 h/day and 24 ± 2°C for the remaining time) were fed the basal diet (HT) or basal diet supplemented with 500 mg/kg resveratrol for 14 consecutive days. Key results Compared with the thermo-neutral group, birds in the HT group had a decreased (P < 0.05) average daily feed intake, average daily gain, villus height, villus height to crypt depth ratio, mRNA concentrations of mucin-2, secreted immunoglobulin A (sIgA), claudin-1,zona occludens-1 and serum concentrations of interferon γ, and increased (P < 0.05) feed to gain ratio, crypt depth, mRNA levels of expression of heat-shock protein (HSP) 70, HSP90, nuclear factor kappa B, mucin-4, claudin-2 and serum concentrations of endotoxin, interleukin (IL)-1β, IL-4, IL-6 and tumour necrosis factor (TNF)-α on Day 3 and Day 14, except for claudin-1 on Day 14 and TNF-α on Day 3 (P > 0.05). Compared with HT group, birds in HT supplemented with resveratrol group decreased (P < 0.05) crypt depth (in jejunum on Day 3, ileum on Day 14), mRNA levels of expression of HSP70, HSP90, nuclear factor kappa B, mucin-4, claudin-2 and serum concentrations of endotoxin, IL-1β, IL-4, IL-6 and TNF-α, and increased (P < 0.05) average daily feed intake (+11%), average daily gain (+22%) and villus height, villus height to crypt depth ratio, mRNA levels of expression of mucin-2, sIgA, claudin-1,zona occludens-1 and serum concentrations of interferon γ, although with few fluctuations between Day 3 and Day 14. Conclusions Dietary supplementation of resveratrol was effective in partially alleviating the adverse effects of heat stress on growth performance and intestinal barrier function in yellow-feather broilers, by restoring the impaired villus-crypt structure, altering the mRNA expression of intestinal HSPs, mucins, sIgA and tight junction-related gene, and inhibiting secretion of pro-inflammation cytokines. Implications Dietary resveratrol supplementation is a considerable nutritional strategy to anti-stress in animal production.

Peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) has gained popularity as a very attractive target for diabetic therapies due to its role in lipid and glucose metabolism. Pharmacological activation of PGC-1α is thought to elicit health benefits. However, this notion has been questioned by increasing evidence, which suggests that insulin resistant is exacerbated when PGC-1α expression is far beyond normal physiological limits and is prevented under the condition of PGC-1α deficiency. This narrative review suggests that PGC-1α, as a master metabolic regulator, exerts roles in insulin sensitivity in a tissue-specific manner and in a physical activity/age-dependent fashion. When using PGC-1α as a target for therapeutic strategies against insulin resistance and T2DM, we should take these factors into consideration. Level of evidence: Level V, narrative review.

The present study is conducted to determinate fatty acids (FA) composition in 3 adipose tissues. Subcutaneous and perirenal adipose tissues were prepared from 24 Ningxiang castrated boars and 24 castrated gilts fattened by a traditional diet for 56 d, respectively. The results showed that the FA profile in the 3 adipose tissues (dorsal subcutaneous adipose [DSA], abdominal subcutaneous adipose [ASA], and perirenal adipose [PA]) differed greatly. In boars, the proportions of oleic acid (c18:1n9c) (P

This study was to investigate the effect of soyabean isoflavones (SIF) on onset of puberty, serum hormone concentration, and gene expression in hypothalamus, pituitary and ovary of female Bama miniature pigs. Fifty five, 35-days old pigs were randomly assigned into 5 treatment groups consisting of 11 pigs per treatment. Results showed that dietary supplementation of varying dosage (0, 250, 500, and 1,250 mg/kg) of SIF induced puberty delay of the pigs with the age of puberty of pigs fed basal diet supplemented with 1,250 mg/kg SIF was significantly higher (p

The effects of dietary supplementation with lysine-yielding Bacillus subtilis on gut morphology, cecal microflora, and expression of intestinal cytokine mRNA of Linwu ducks (a Chinese local domesticated duck) were investigated after a 63-d feeding trial. Two hundred forty 1-d-old female ducklings were randomly assigned to 1 of 4 dietary treatments, and there were 60 birds per treatment replicated 6 times with 10 birds each. The experimental diets consisted of a basal diet containing no antibiotic or growth promoters, the basal diet supplemented with 150 mg Aureomycin per kg feed, and 2 others supplemented with 5.0 × 108 and 5.0 × 1010 cfu B. subtilis/kg feed, respectively. Birds fed the diets supplemented with B. subtilis had increased (P < 0.001) villi height in the jejunum and ileum and reduced (P < 0.001) crypt depth in the jejunum compared with birds fed the control diet. Greatest (P < 0.01) villus height:crypt depth ratio was obtained with the jejunum and ileum of birds fed the diet supplemented with 5.0 × 1010 cfu B. subtilis/kg feed. Dietary supplementation with B. subtilis resulted in increased (P < 0.05) cecal counts of Megasphaera elsdenii. Greater cecal Lactobacillus counts were obtained in ducks fed the diets supplemented with Aureomycin and B. subtilis. Ducks fed the diets supplemented with B. subtilis showed suppressed (P < 0.001) expression of interleukin (IL)-1β, IL-2, and IL-10 in their jejunum. When compared with the control, the expression of interferon (IFN)-γ in the jejunum of ducks fed the diet supplemented with 5.0 × 108 cfu B. subtilis/kg feed was significantly increased (P < 0.05). Dietary supplementation with B. subtilis or Aureomycin reduced (P < 0.001) ileal expression of IL-10. When compared with the control, the expression of IL-2 and IL-8 in the ileum of ducks fed the diet supplemented with 5.0 × 1010 cfu B. subtilis/kg feed showed a significant decrease (P < 0.05). In conclusion, dietary supplementation with lysine-yielding B. subtilis improved gut morphology, increased the population of beneficial gut microflora, and stimulated increased intestinal immune response of Linwu ducks.

Polyamines are necessary for normal integrity and the restitution after injury of the gastrointestinal epithelium. The objective of this study was to investigate the effects of oral administration of putrescine and proline during the suckling period on epithelial restitution after early weaning in piglets. Eighteen neonatal piglets (Duroc × Landrace × Large Yorkshire) from 3 litters (6 piglets per litter) were assigned to 3 groups, representing oral administration with an equal volume of saline (control), putrescine (5 mg/kg BW), and proline (25 mg/kg BW) twice daily from d 1 to weaning at 14 d of age. Plasma and intestinal samples were obtained 3 d after weaning. The results showed that oral administration of putrescine or proline increased the final BW and ADG of piglets compared with the control (P < 0.05). Proline treatment decreased plasma D-lactate concentration but increased the villus height in the jejunum and ileum, as well as the percentage of proliferating cell nuclear antigen (PCNA) positive cells and alkaline phosphatase (AKP) activity in the jejunal mucosa (P < 0.05). The protein expressions for zonula occludens (ZO-1), occludin, and claudin-3 (P < 0.05) but not mRNA were increased in the jejunum of putrescine- and proline-treated piglets compared with those of control piglets. The voltage-gated K+ channel (Kv) 1.1 protein expression in the jejunum of piglets administrated with putrescine and the Kv1.5 mRNA and Kv1.1 protein levels in the ileum of piglets administrated with proline were greater than those in control piglets (P < 0.05). These findings indicate that polyamine or its precursor could improve mucosal proliferation, intestinal morphology, as well as tight junction and potassium channel protein expressions in early-weaned piglets, with implications for epithelial restitution and barrier function after stress injury.

PCV2 is highly pathogenic, however, its effect on the serum amino acids profile is unknown. This study was conducted to explore the profile of amino acids in the serum in porcine circovirus type 2 (PCV2) infected mice. The serum levels of amino acids were detected with isotope dilution liquid chromatography-mass spectrometry methods at 3, 7, 10 and 14 days post infection (DPI). Meanwhile, the expression of seven amino acids transporters (SLC6A14, SLC6A20, SLC7A5, SLC7A6, SLC7A7, SLC7A8, SLC7A9) in the jejunum was analyzed by reverse transcription polymerase chain reaction (RT-PCR) at 3 and 7 DPI. Serum PCV2 load was also analyzed by quantitative PCR at 3, 7, 10 and 14 DPI. Serum levels of most amino acids, such as Pro, Orn, and Met, significantly (P < 0.05) increased at 3 DPI. However, most amino acids, including Asp, Sar, Arg, Hyl, Pro, Lys, Val, Ile and Leu, significantly (P < 0.05) decreased at 7 DPI. There was no significant difference for most amino acids at 10 and 14 DPI. PCV2 infection significantly (P < 0.05) decreased expression of SLC7A5 and SLC7A6 at 7 DPI. In conclusion, PCV2 infection affects the profile of amino acids in the serum and the expression of amino acids transporters in the intestine.

The mycotoxin deoxynivalenol (DON) is a food contaminant that leads to reduced feed intake and reduced BW gain, as well as organ impairment. On the other hand, antimicrobial peptides have been shown to have positive effects on growth performance, nutrient digestibility, and immune function. The purpose of this study was to investigate the protective effects of composite antimicrobial peptides (CAP) on piglets challenged with DON. After a 7-d adaptation period, 28 individually housed piglets (Duroc × Landrace × Large Yorkshire) weaned at 28 d of age were randomly assigned to receive 1 of 4 treatments (7 pigs/treatment): negative control, basal diet (NC), basal diet + 0.4% CAP (CAP), basal diet + 4 mg/kg DON (DON), and basal diet + 4 ppm DON + 0.4% CAP (DON + CAP). On d 15 and 30 after the initiation of treatment, blood samples were collected for the determination of blood profile. Piglets were monitored for 30 d to assess performance and then were slaughtered to obtain organs for the determination of the relative weight of organs. The results showed that dietary supplementation with DON decreased (P 0.05) on other relative weights of viscera, except the relative weight of the gallbladder, but the diamine oxidase activity in the liver decreased in DON-treated piglets (P 0.05) between the DON + CAP treatment and the other treatments. The DON treatment decreased the numbers of red blood cells and platelets, as well as the serum catalase concentrations, and decreased the serum concentrations of H2O2, maleic dialdehyde, and nitric oxide (P < 0.05). The numbers of platelets and thrombocytocrit, as well as the serum concentrations of catalase, were greater, whereas the maleic dialdehyde concentrations were decreased, in both the CAP and DON + CAP treatments compared with the other treatments (P < 0.05). Compared with the control treatment, DON decreased peripheral lymphocyte proliferation on d 15, whereas supplementation with CAP increased it on d 15 and 30 (P < 0.05). These findings indicate that CAP could improve feed efficiency, immune function, and antioxidation capacity and alleviate organ damage, and thus, it has a protective effect in piglets challenged with DON.

Periostin, also called osteoblast-specific factor 2, is an important regulator of bone, cardiac development, and wound healing. A recent study revealed that periostin plays an important role in tumor development and is upregulated in a wide variety of cancers. However, little is known about periostin in swine. Therefore, the cDNA sequence of the porcine periostin gene was obtained by rapid amplification of cDNA ends (RACE). One C/T single nucleotide polymorphism anchored in intron 9 was identified and genotyped by PCR-RFLP-HaeIII. In Daweizi, Shaziling, Ningxiang, Taoyuan, Wuzhishan, Landrace, and Yorkshire pigs, the C allele was dominant, while the T allele was dominant in the Duroc pig. Quantitative PCR analysis showed that the periostin gene was expressed in all examined tissues from 25-day-old Shaziling and Yorkshire piglets, with mRNA expression in the longissimus dorsi muscle being the highest in these two breeds, and that in the kidney and lungs being the lowest. There was a significant difference in periostin gene expression in the intestines, heart, and spleen (P < 0.05). These findings might contribute to our understanding of the function of periostin in swine.

L-Glutamine is a nutritionally semi-essential amino acid for proper growth in most cells and tissues, and plays an important role in the determination and guarding of the normal metabolic processes of the cells. With the help of transport systems, extracellular L-glutamine crosses the plasma membrane and is converted into alpha-ketoglutarate (AKG) through two pathways, namely, the glutaminase (GLS) I and II pathway. Reversely, AKG can be converted into glutamine by glutamate dehydrogenase (GDH) and glutamine synthetase (GS), or be converted into CO2 via the tricarboxylic acid (TCA) cycle and provide energy for the cells. Different steps of glutamine metabolism (the glutamine-AKG axis) are regulated by several factors, rendering the glutamine-AKG axis a potential target to counteract cancer. Moreover, intracellular glutamine plays an important role in cellular homeostasis not only as a precursor for protein synthesis, but also for its nutritional roles in cell growth, lipid metabolism, insulin secretion, and so on. The main objective of this review is to highlight the metabolic pathways of glutamine to AKG, with special emphasis on nutritional and therapeutic use of glutamine-AKG axis to improve the health and well-being of animals and humans.

Chitosan oligosaccharide (COS) is a degradation product of chitosan with antioxidative, anti-inflammatory, and antibacterial effects. This study was conducted to investigate the effects of dietary COS on the intestinal inflammatory response and the calcium-sensing receptor (CaSR) and nuclear transcription factor kappa B (NF-κB) signaling pathways that may be involved using a lipopolysaccharide (LPS)-challenged piglet model. A total of 40 weaned piglets were used in a 2 × 2 factorial design; the main factors were dietary treatment (basal or 300 μg/kg COS) and inflammatory challenge (LPS or saline). On the morning of days 14 and 21 after the initiation of treatment, the piglets were injected intraperitoneally with Escherichia coli LPS at 60 and 80 μg/kg body weight or the same amount of sterilized saline, respectively. Blood and small intestine samples were collected on day 14 or 21, respectively. The results showed that piglets challenged with LPS have a significant decrease in average daily gain and gain:feed and histopathological injury in the jejunum and ileum, whereas dietary supplementation with COS significantly alleviated intestinal injury induced by LPS. Piglets fed the COS diet had lower serum concentrations of tumor necrosis factor alpha (TNF-α), interleukin (IL) 6, and IL-8 as well as lower intestinal abundances of pro-inflammatory cytokine mRNA but higher anti-inflammatory cytokine mRNA compared with piglets fed the basal diet among LPS-challenged piglets (p < 0.05). Dietary COS increased intestinal CaSR and PLCβ2 protein expressions in both saline- and LPS-treated piglets, but decreased p-NF-κB p65, IKKα/β, and IκB protein expressions in LPS-challenged piglets (p < 0.05). These findings indicate that COS has the potential to reduce the intestinal inflammatory response, which is concomitant with the activation of CaSR and the inhibition of NF-κB signaling pathways under an inflammatory stimulus.

Chitosan oligosaccharide (COS) has a blood glucose lowering effect in diabetic rats and is widely used as a dietary supplement. However, the effect of COS on the offspring of supplemented mothers is unknown. This experiment investigates the effect of supplementing sows during gestation and lactation on the levels of plasma glucose on suckling piglets. From day 85 of gestation to day 14 of lactation, 40 pregnant sows were divided into two treatment groups and fed either a control diet or a control diet containing 30mgCOS/kg. One 14 day old piglet per pen was selected to collect plasma and tissue (8pens/diet). Performance, hepatic gluconeogenesis genes and proteins expression, amino acids contents in sow milk, hepatic glycogen and free fatty acid were determined. Results showed that supplementation of the maternal diet with COS improved daily gain and weaning weight (P

Longlin pig is a native breed of Fujian province in China. It is the first time that the complete mitochondrial genome sequence of Longlin pig is reported in this work, which is determined through the PCR-based method. The total length of the mitognome is 16,699 bp, which contains 1 control region (D-loop region), 2 ribosomal RNA genes, 13 PCGs and 22 tRNA genes. The total base composition of Longlin pig mitochondrial genome is 34.67% for A, 26.21% for C, 25.80% for T and 13.33% for G, in the order A>C>T>G. The complete mitochondrial genome of Longlin pig provides an important data in genetic mechanism and the evolution genomes.

In this study, the complete mitochondrial genome sequence of Diannan small-ear pig in Yunnan Province was firstly reported, which was determined through polymerase chain reaction (PCR) method. The total length of mitochondrial genome of Diannan small-ear pig was 16720 bp, including 34.77% A, 26.18% C, 25.81% T and 13.24% G, and in the order A > C > T > G. Mitochondrial genome contained a major non-coding control region (D-Loop region), 2 ribosomal RNA genes, 13 protein-coding genes (PCGs) and 22 transfer RNA genes. The mitochondrial genome of Diannan small-ear pig provides an important data set for the study on genetic mechanism.

The aim of this study was to investigate whether supplementation with chitosan (COS) could reduce diarrhea and to explore how COS alleviates intestinal inflammation in weaned pigs. Thirty pigs (Duroc×Landrace×Yorkshire, initial BW of 5.65±0.27) weaned at age 21 d were challenged with enterotoxigenic Escherichia coli during a preliminary trial period, and then divided into three treatment groups. Pigs in individual pens were fed a corn-soybean meal diet, that contained either 0 (control), 50 mg/kg chlortetracycline, or 300 mg/kg COS for 21 days. The post-weaning diarrhea frequency, calprotectin levels and TLR4 protein expression were decreased (P

Continuous synthesis and breakdown or remodeling of proteins (also called protein turnover) is a principal characteristic of protein metabolism. During animal production, the net differences between synthesis and breakdown represent the actual marketable muscle foods. Because protein synthesis is highly endergonic and protein breakdown is metabolically energy dependent, the efficiency of production can be markedly enhanced by lower muscle protein breakdown rates. Various methodological approaches to study protein synthesis and breakdown, with particular emphasis toward food-producing animals, are presented. These include whole-animal tracer amino acid infusion in vivo, quantifying marker amino acid release from muscle proteins, and in vitro amino acid release-based methodologies. From such methods, protein synthesis rates and protein breakdown rates (mass units/time) may be obtained (Helene et al. 2002). The paper briefly reviews the methodology developed over the past 30 years to study protein turnover.

This study was conducted to investigate how chitosan (COS) affects intestinal mucosal barrier function and to further explain mechanisms of COS on growth performance. Thirty piglets, weaned at 21 days of age, were challenged with enterotoxigenic Escherichia coli during preliminary trial period. Three groups of Piglets in individual pens were fed a corn-soybean meal diet containing no addition, 50 mg/kg chlortetracycline, or 300 mg/kg COS for 21 days. Jejunal morphology and histology were analyzed under light microscope. The concentrations of occludin proteins were determined by western blot. Immunohistochemistry assays were used to determine secretory immunoglobulin (sIgA) level. Real-time PCR was used to detect Toll-like receptor 4 (TLR4) and Claudin-1 in jejunal mucosa. Feeding COS or chlortetracycline reduced (P0.05) feed conversion ratio. Villus length, villus length/crypt depth, and goblet cells, were increased (P0.05), but villus width and crypt depth were decreased (P0.05) in both COS and chlortetracycline groups. Intraepithelial lymphocytes were higher (P0.05) in the COS group than both chlortetracycline and control groups. Occludin protein expression was increased (P0.01) in the COS group, but was decreased (P0.05) in the chlortetracycline group. Expression of sIgA protein was higher (P0.05) in the COS group than both control and chlortetracycline groups, however TLR4 mRNA expression was decreased (P0.05) in both COS and chlortetracycline groups. There was no difference in expression of claudin-1 among the three groups. In conclusion, chitosan and the antibiotic have similar effects in promoting piglet growth and reducing intestinal inflammation, but different effects on intestinal mucosal barrier function. This indicates that chitosan can replace chlortetracycline as a feed additive for piglets.

Wuyi Black pig is a native breed of Fujian province in China. It is the first time that the complete mitochondrial genome sequence of Wuyi Black pig is reported in this work, which is determined through the PCR-based method. The total length of the mitognome is 16,709 bp, which contains 2 ribosomal RNA genes, 22 tRNA genes, 13 PCGs and 1 control region (D-loop region). The total base composition of Wuyi Black pig mitochondrial genome is 34.67% for A, 26.20% for C, 25.81% for T and 13.33% for G, in the order A > C > T > G. The complete mitochondrial genome of Wuyi Black pig provides an important data in genetic mechanism and the evolution genomes.

This study was conducted to determine the effect of dietary supplementation of a low dose of chito-oligosaccharide (COS) on intestinal morphology, immune response, antioxidant capacity, and barrier function in weaned piglets. A total of 120 weaned pigs (21 d of age; 7.86 ± 0.22 kg average BW) were randomly assigned (6 pens/diet; 10 pigs/pen) to 2 dietary treatments consisting of a basal diet (negative control) or the basal diet supplemented with COS (30 mg/kg) for a 14-d period. Six randomly selected piglets from each treatment were killed for blood and tissue sampling. No significant differences were observed in ADG, ADFI, and G:F between treatment and the control group. Piglets fed the COS-supplemented diet had greater ( < 0.05) stomach pH than those fed the control diet on d 14 postweaning. Dietary supplementation with COS reduced villus height ( < 0.05) and villus height:crypt depth ( < 0.05) in the ileum. Dietary COS supplementation tended to reduce villus height in the duodenum ( = 0.065) and jejunum ( = 0.058). There was no effect on crypt depth in the intestinal segments of treatment group. Piglets fed the COS-supplemented diet increased ( < 0.05) the number of intraepithelial lymphocytes in duodenum or jejunum and goblet cells of ileum. However, COS decreased ( < 0.05) the number of intraepithelial lymphocytes in ileum of weaned piglets. The concentrations of IL-10 (duodenum, jejunum, and ileum) and secretory immunoglobulin (SIgA; duodenum and ileum) were higher in piglets fed the COS-supplemented diet compared with control ( < 0.05). Dietary COS supplementation reduced ( < 0.05) the concentration of total antioxidant capacity and superoxide dismutase of the jejunum or ileum. The mRNA expression of occludin in the ileum and ZO-1 in jejunum and ileum had a significant change in piglets fed the COS-supplemented diet compared with the control group ( < 0.05). In conclusion, these results indicated that dietary COS supplementation at 30 mg/kg had no effects on promoting growth performance and tended to reduce villus height in the duodenum or jejunum of weaned piglets. The results further showed that supplemental COS at this level may cause an immune and oxidative stress response in small intestine and have compromised the intestinal barrier integrity in weaned piglets. The research will provide guidance on the low dosage of COS supplementation on weaning pigs.