Qiuping Guo

Effects of dietary nutrient levels on microbial community composition and diversity in the ileal contents of pregnant Huanjiang mini-pigs

The mammalian gut microbiota influences various metabolic and physiological processes. Substantial metabolic changes occur during a healthy pregnancy that may be related to microbiota composition dynamics. However, the effect of diet on intestinal microbiota composition and diversity during pregnancy remains unclear. We examined the ileal contents of Huanjiang mini-pigs at two pregnancy stages to determine the effects of dietary nutrient levels on such microbial communities. Animals received either a higher-nutrient (HN) diet formulated to meet US National Research Council requirements or a lower-nutrient (LN) diet that met the Chinese National Feeding Standard recommendations. On day 45 or 75 of pregnancy, sows were euthanized and their ileal contents sampled. Experimental diet and pregnancy stage did not affect ileal bacterial richness or diversity, as determined by Chao1 and ACE species richness measures and Shannon and Simpson indices, respectively. The phyla Firmicutes and Proteobacteria, accounting for 69.99–85.44% and 5.82–15.17% of the total reads, respectively, predominated regardless of diet. At the genus level, diet significantly affected the abundance of Lactobacillus species, which was greater in pigs given HN feed (P < 0.05), but had little impact on that of Megasphaera species (P = 0.096). Pregnancy stage had a minimal effect on Proteobacteria numbers (P = 0.053). The number of bacteria of the phylum Firmicutes and genus Lactobacillus decreased, while that of the phylum Proteobacteria, family Enterobacteriaceae, and genus Bacteroides increased between days 45 and 75 of pregnancy. Of the short-chain fatty acids (SCFAs) measured, only propionate levels changed significantly, with higher concentrations observed on day 45 than on day 75. Our findings indicate that Firmicutes and Proteobacteria dominate pregnant sow ileal bacterial profiles. Excepting a tendency for the number of Proteobacteria to increase as pregnancy progressed, pregnancy stage and diet had little effect on ileal microbiotic composition and diversity and luminal SCFA concentrations.

Alteration of inflammatory cytokines, energy metabolic regulators, and muscle fiber type in the skeletal muscle of postweaning piglets

This study was conducted to determine the alterations of inflammatory cytokines, energy metabolic regulators, and muscle fiber type in the LM of the piglets postweaning. Crossbred piglets (Landrace × Large White) weaned at 14 d age were randomly selected from 8 litters and slaughtered at 0 (W0), 1 (W1), 3 (W3), 5 (W5), or 7 (W7) days postweaning. The glycogen content, free glucose concentration, and enzyme activities, including ATPase (Na/K, Ca/Mg), creatine kinase, and lactic dehydrogenase (LDH), were detected in the skeletal muscle tissue. Concentrations of proinflammatory cytokines, including IL-1β, IL-6, and tumor necrosis factor-α (TNF-α), and anti-inflammatory cytokines, including IL-10 and transforming growth factor-β1 (TGF-β1), were measured in serum. The mRNA abundance of the above cytokines, energy metabolic regulators, and muscle fiber type related genes were determined via real-time quantitative PCR analysis. The adenosine monophosphate-activated protein kinase α (AMPKα) signaling was measured by Western blot analysis. Our results showed ATPase activities were lower on W7 d but LDH activity was higher on W3 d after weaning ( < 0.05). Serum TNF-α concentration was markedly increased on W1 d, then returned to the value of preweaning ( < 0.05), and almost all the values of inflammatory cytokines were reduced to a low point on W5 d after weaning. Additionally, the IL-6 mRNA abundance was upregulated during W3 to W7 d, but cytokine TNF-α was upregulated just on W7 d ( < 0.05). The mRNA abundance of AMPKα and uncoupling protein (UCP) 3 were both higher on W1 and W3 d, and UCP2 was higher on W7 d postweaning ( < 0.05). Myosin heavy chain (MyHC) I and MyHC IIx-type fibers were enhanced on W1 d, then returned to the value of preweaning, and the MyHC IIb-type fiber was significantly increased on W5 and W7 d ( < 0.05). Meanwhile, the value of P-AMPKα/T-AMPKα increased on W3 d postweaning ( < 0.05) compared with that on W0 d. These results indicate that weaning stress induced inflammation in skeletal muscle tissue during at least 7 d postweaning. It upregulated the expression of proinflammatory cytokines, which then stimulated the AMPKα and UCP involved in energy metabolism events, accompanied by significant alterations in muscle fiber type.

Effects of Low-Protein Diets Supplemented with Branched-Chain Amino Acid on Lipid Metabolism in White Adipose Tissue of Piglets

This study evaluated the effect of branched-chain amino acid (BCAA) supplementation in low-protein diets on lipid metabolism in dorsal subcutaneous adipose (DSA), abdominal subcutaneous adipose (ASA), and perirenal adipose (PRA) tissues. A total of 24 piglets were allotted to four treatments, and each group was fed the adequate protein (AP) diet, low-protein (LP) diet, LP diet supplemented with BCAA (LP + B), or LP diet supplemented with twice BCAA (LP + 2B). Serum concentrations of leptin in the BCAA-supplemented treatments were higher (P < 0.01) than in the AP treatment, but lower (P < 0.01) than in the LP treatment. In DSA, the mRNA and protein levels for lipogenic-related genes were highest in the LP treatment and lowest in the LP + 2B treatment. However, in ASA and PRA, the expression levels for those genes were significantly elevated in the LP + 2B treatment. In conclusion, BCAA supplementation could alter the body fat condition, and this effect was likely modulated by the expression of lipid metabolic regulators in DSA, ASA, and PRA in a depot-specific manner.

Supplementation of branched-chain amino acids in protein-restricted diets modulates the expression levels of amino acid transporters and energy metabolism associated regulators in the adipose tissue of growing pigs

This experiment was conducted to investigate the effects of branched-chain amino acids (BCAA) supplemented in protein-restricted diets on the growth performance and the expression profile of amino acid transporters and energy metabolism related regulators in the white adipose tissue (WAT) of different regional depots including dorsal subcutaneous adipose (DSA) and abdominal subcutaneous adipose (ASA). A total of 24 crossbred barrows (7.40 ± 0.70 kg) were randomly divided into 4 groups and were fed the following isocaloric diets for 33 days: 1) a recommended adequate protein diet (AP, 20% CP, as a positive control); 2) a low protein diet (LP, 17% CP); 3) the LP diet supplemented with BCAA (LP + B, 17% CP) to reach the same level of the AP diet group; 4) the LP diet supplemented with 2 times the amount of BCAA (LP + 2B, 17% CP). The daily gain and daily feed intake of the LP diet group were the lowest among all the treatments (P < 0.01). The feed conversion was improved markedly in the group of LP + B compared with the LP diet group (P < 0.05). No significant difference was noted for the serum biochemical parameter concentrations of glucose, triglyceride, nonesterified fatty acid and insulin among the groups (P > 0.05). Moreover, BCAA supplementation down-regulated the expression levels of amino acid transporters including L-type amino acid transporter 1 and sodium-coupled neutral amino acid transporter 2 in DSA, but up-regulated the expression level of L-type amino acid transporter 4 in ASA (P < 0.05). Meanwhile, the energy sensor AMP-activated protein kinase α was activated in the DSA of pigs fed LP diet and in the ASA of the pigs fed AP or LP + 2B diets (P < 0.05). The mRNA expression profile of the selected mitochondrial component and mitochondrial biogenesis associated regulators in DSA and ASA also responded differently to dietary BCAA supplementation. These results suggested that the growth performance of growing pigs fed protein restricted diets supplemented with BCAA could catch up to that of the pigs fed AP diets. The results also partly demonstrated that the regulation mechanisms of BCAA are different in the adipose tissues of different depots.

Dietary proline supplementation alters colonic luminal microbiota and bacterial metabolite composition between days 45 and 70 of pregnancy in Huanjiang mini-pigs

BackgroundPregnancy is associated with important changes in gut microbiota composition. Dietary factors may affect the diversity, composition, and metabolic activity of the intestinal microbiota. Among amino acids, proline is known to play important roles in protein metabolism and structure, cell differentiation, conceptus growth and development, and gut microbiota re-equilibration in case of dysbiosis.ResultsDietary supplementation with 1% proline decreased (P < 0.05) the amounts of Klebsiella pneumoniae, Peptostreptococcus productus, Pseudomonas, and Veillonella spp. in distal colonic contents than that in the control group. The colonic contents of Butyrivibrio fibrisolvens, Bifidobacterium sp., Clostridium coccoides, Clostridium coccoides-Eubacterium rectale, Clostridium leptum subgroup, Escherichia coli, Faecalibacterium prausnitzii, Fusobacterium prausnitzii, and Prevotella increased (P < 0.05) on d 70 of pregnancy as compared with those on d 45 of pregnancy. The colonic concentrations of acetate, total straight-chain fatty acid, and total short-chain fatty acids (SCFA) in the proline-supplemented group were lower (P < 0.05), and butyrate level (P = 0.06) decreased as compared with the control group. Almost all of the SCFA displayed higher (P < 0.05) concentrations in proximal colonic contents on d 70 of pregnancy than those on d 45 of pregnancy. The concentrations of 1,7-heptyl diamine (P = 0.09) and phenylethylamine (P < 0.05) in proximal colonic contents were higher, while those of spermidine (P = 0.05) and total bioamine (P = 0.06) tended to be lower in the proline-supplemented group than those in the control group. The concentrations of spermidine, spermine, and total bioamine in colonic contents were higher (P < 0.05) on d 70 of pregnancy than those measured on d 45 of pregnancy. In contrast, the concentration of phenylethylamine was lower (P < 0.05) on d 70 than on d 45 of pregnancy.ConclusionThese findings indicate that L-proline supplementation modifies both the colonic microbiota composition and the luminal concentrations of several bacterial metabolites. Furthermore, our data show that both the microbiota composition and the concentrations of bacterial metabolites are evolving in the course of pregnancy. These results are discussed in terms of possible implication in terms of luminal environment and consequences for gut physiology and health.

Alteration of muscle fiber characteristics and the AMPK-SIRT1-PGC-1α axis in skeletal muscle of growing pigs fed low-protein diets with varying branched-chain amino acid ratios

There mainly exists four major myosin heavy chains (MyHC) (i.e., I, IIa, IIx, and IIb) in growing pigs. The current study aimed to explore the effects of low-protein diets supplemented with varying branched-chain amino acids (BCAAs) on muscle fiber characteristics and the AMPK-SIRT1-PGC-1α axis in skeletal muscles. Forty growing pigs (9.85 ± 0.35 kg) were allotted to 5 groups and fed with diets supplemented with varying leucine: isoleucine: valine ratios: 1:0.51:0.63 (20% crude protein, CP), 1:1:1 (17% CP), 1:0.75:0.75 (17% CP), 1:0.51:0.63 (17% CP), and 1:0.25:0.25 (17% CP), respectively. The skeletal muscles of different muscle fiber composition, that is, longissimus dorsi muscle (LM, a fast-twitch glycolytic muscle), biceps femoris muscle (BM, a mixed slow- and fast-twitch oxido-glycolytic muscle), and psoas major muscle (PM, a slow-twitch oxidative muscle) were collected and analyzed. Results showed that relative to the control group (1:0.51:0.63, 20% CP), the low-protein diets with the leucine: isoleucine: valine ratio ranging from 1:0.75:0.75 to 1:0.25:0.25 especially augmented the mRNA and protein abundance of MyHC I fibers in BM and lowered the mRNA abundance of MyHC IIb particularly in LM (P < 0.05), with a concurrent increase in the activation of AMPK and the mRNA abundance of SIRT and PGC-1α in BM (P < 0.05). The results reveal that low-protein diets supplemented with optimal BCAA ratio, i.e. 1:0.75:0.75-1:0.25:0.25, induce muscle more oxidative especially in oxido-glycolytic skeletal muscle of growing pigs. These effects are likely associated with the activation of the AMPK-SIRT1-PGC-1α axis.

β-hydroxy-β-methyl butyrate is more potent than leucine in inhibiting starvation-induced protein degradation in C2C12 myotubes

Leucine (Leu) and its metabolites α-ketoisocaproate (KIC) and β-hydroxy-β-methyl butyrate (HMB) are potent regulators of protein turnover. The aim of this study was to compare the inhibitory effects of Leu, KIC, and HMB on protein degradation and to investigate the mechanisms involved. The results showed that the inhibitory effect of HMB (0.38 ± 0.04) was more potent than that of Leu (0.76 ± 0.04) and KIC (0.56 ± 0.04, P < 0.01), and was significantly abolished in the presence of LY294002 (1.48 ± 0.02) and rapamycin (1.96 ± 0.02, P < 0.01). In the presence of insulin, the inhibitory effect of HMB (0.34 ± 0.03) was still more effective than that of Leu (0.60 ± 0.04) and KIC (0.57 ± 0.08, P < 0.05). Interestingly, LY294002 treatment markedly attenuated the effect of HMB, while rapamycin treatment failed to exert the same effect. Thus, HMB appears to be more potent than Leu and KIC in inhibiting protein degradation in the absence or presence of insulin, and this inhibitory effect may be dependent on PI3K/Akt signaling pathway regardless of insulin, and mTOR signaling was only involved in this effect of HMB in the absence of insulin.

The Protein and Energy Metabolic Response of Skeletal Muscle to the Low-Protein Diets in Growing Pigs

This study was conducted to determine the effect of low-protein diets on protein and energy metabolism in skeletal muscle, and to elucidate the underlying mechanism. A total of 18 growing pigs (average initial body weight = 36.47 kg) were individually penned and assigned to three treatments; each treatment was fed one of three diets containing either 18%, 15%, or 12% CP. The results showed that reducing dietary CP contents decreased (P < 0.05) the weight of half Longissimus dorsi (LD) muscle and serum concentration of insulin-like growth factor 1 (IGF-1). Compared with the 18% and 15% CP treatments, the 12% CP treatment suppressed (P < 0.05) the components of mammalian target of rapamycin complex 1 (mTORC1) pathway, but upregulated (P < 0.05) the mRNA levels for proteolysis-related genes, and concomitantly caused an increase (P < 0.05) in the percentage of apoptotic cells in LD muscle. Along with lower (P < 0.05) AMP/ATP ratio and greater (P < 0.05) energy charge value in LD muscle of the 12% CP treatment, there was a concurrent decrease (P < 0.05) in the proteins for AMP-activated protein kinase α (AMPKα) pathway. Likewise, these results were also observed in the Biceps femoris muscle with slightly different degree of impacts. These results indicate that the retardation effect of low-protein supply on muscle growth of growing pigs could be likely regulated by inhibiting IGF-1/mTORC1 protein synthesis cascade, along with strong alterations in energy status and AMPKα pathway.

Taurine is Involved in Energy Metabolism in Muscles, Adipose Tissue, and the Liver

Energy metabolism is a basic and general process, by which the body acquires and uses energy to maintain normal function, and taurine plays a vital role in energy metabolism. Taurine deficiency may cause a weak energy metabolism and energy metabolism dysfunction. Taurine biosynthetic ability is limited, and its supplementation in the diet can strengthen energy metabolism in muscle performance, cardiac function, liver activity, and adipose tissue. Combining taurine with other drugs may have a superior effect in energy metabolism. In many metabolic disorders, taurine, or the combination of taurine with other drugs, also functions as a repair treatment for damaged tissues, and acts as a promoter for the balance of energy metabolism. The present study discusses the potential roles of taurine in energy metabolism.

Low-protein diet improves meat quality of growing and finishing pigs through changing lipid metabolism, fiber characteristics, and free amino acid profile of the muscle

The objective of the study was to investigate the effect of feeding reduced CP, AA-supplemented diets on meat quality in growing and finishing pigs as well as the related mechanism. In experiment 1, 18 growing pigs (36.5 kg BW) were assigned randomly and fed 1 of 3 corn-soybean meal diets containing either 18% CP (normal protein, NP), 15% CP (low protein, LP), or 12% CP (very low protein, VLP). In experiment 2, 18 finishing pigs (62.3 kg BW) were allotted randomly into 1 of the following diets: 16% CP (NP), 13% CP (LP), or 10% CP (VLP). In both experiments, the LP and VLP diets were supplemented with crystalline AA to achieve equal content of standardized ileal digestible lysine, methionine, threonine, and tryptophan. At the end of each experiment, all pigs were slaughtered to collect longissimus dorsi muscle (LM) samples. Samples were used for determining meat quality, intramuscular fat (IMF) content, fatty acid composition, free AA profile, and expression of genes for myosin heavy chain isoforms. Results showed that growing and finishing pigs fed the LP diets increased (P < 0.05) redness value of LM, while finishing pigs fed the LP and VLP diets decreased (P < 0.05) the shear force values. Compared with the NP diet, growing and finishing pigs fed lower CP diets had higher (P < 0.05) contents of IMF and MUFA, and lower (P < 0.05) contents of PUFA. Besides, higher (P < 0.05) expression levels of type I and/or IIa muscle fibers were observed in LP diet-fed growing and finishing pigs, and greater concentrations of taurine and tasty AA in VLP diet-fed growing and finishing pigs. Taken together, our results indicate that low-protein diets could positively affect meat quality of growing and finishing pigs, and likely through regulation of IMF content and fatty acid composition, fiber characteristics, and free AA profile in the muscle.