Genome-wide association study identified five quantitative trait loci and two candidate genes for digestive traits in Suhuai pigs.

This work aimed to identify markers and candidate genes underlying porcine digestive traits. In total, 331 pigs were genotyped by 80 K Chip data or 50 K Chip data. For apparent neutral detergent fiber digestibility, a total of 19 and 21 candidate single nucleotide polymorphisms (SNP) were respectively identified using a genome-wide efficient mixed-model association algorithm and linkage-disequilibrium adjusted kinship. Among them, three quantitative trait locus (QTL) regions were identified. For apparent acid detergent fiber digestibility, a total of 16 and 17 SNPs were identified by these two methods, respectively. Of these, three QTL regions were also identified. Moreover, two candidate genes (MST1 and LATS1), which are functionally related to intestinal homeostasis and health, were detected near these significant SNPs. Taken together, our results could provide a basis for deeper research on digestive traits in pigs.

Identification of Gut Microbiota Affecting Fiber Digestibility in Pigs.

Dietary fiber plays an important role in porcine gut health and welfare. Fiber is degraded by microbial fermentation in the intestine, and most gut microbiota related to fiber digestibility in pigs are worth pursuing. The aim of this study was to identify gut microbiota associated with the apparent total tract digestibility (ATTD) of neutral detergent fiber (NDF) and of acid detergent fiber (ADF) in pigs. Large phenotypic variations in the ATTD of NDF and of ADF were separately found among 274 Suhuai pigs. Microbial community structures were significantly different between high and low fiber digestibility groups. Fourteen genera separately dominated the communities found in the high ATTD (H-AD) of NDF and ADF samples and were in very low abundance in the low ATTD (L-AD) of NDF and ADF samples. In conclusion, norank_f__Bacteroidales_S24-7_group (p < 0.05), Ruminococcaceae_UCG-005 (p < 0.05), unclassified_f__Lachnospiraceae (p < 0.05), Treponema_2 (p < 0.01), and Ruminococcaceae_NK4A214_group (p < 0.01) were the main genera of gut microbiota affecting the ATTD of NDF in pigs. Christensenellaceae_R-7_group (p < 0.01), Treponema_2 (p < 0.05), Ruminococcaceae_NK4A214_group (p < 0.05), Ruminococcaceae_UCG-002 (p < 0.05), and [Eubacterium]_coprostanoligenes_group (p < 0.05) were the main genera of gut microbiota affecting the ATTD of ADF in pigs. The most important functions of the above different potential biomarkers were: carbohydrate transport and metabolism, general function prediction only, amino acid transport and metabolism, cell wall/membrane/envelope biogenesis, translation, transcription, replication, energy production and conversion, signal transduction mechanisms, and inorganic ion transport and metabolism. The most important metabolic pathways of the above different potential biomarkers were: membrane transport, carbohydrate metabolism, amino acid metabolism, replication and repair, translation, cell motility, energy metabolism, poorly characterized, nucleotide metabolism, metabolism of cofactors and vitamins, and cellular processes and signaling.

Characteristics of Gut Microbiota in Sows and Their Relationship with Apparent Nutrient Digestibility.

The gut microbiota plays important roles in animal health and nutrient digestibility. The characteristics of gut microbiota population in grower pigs and their correlation with apparent nutrient digestibility were assessed in previous study. Here we studied characteristics of intestinal microbiota of sows and analyzed their relationships with apparent nutrient (ether extract (EE), crude protein (CP), crude fiber (CF), neutral detergent fiber (NDF) and acid detergent fiber (ADF)) digestibility. Firmicutes and Bacteroidetes were the most dominant phyla, approximately 73% of the total sequences. Treponema, Oscillibacter and Lactobacillus were the most dominant generas, more than 49% of the total sequences. The microbiota of sows clustered separately from the microbiota of grower pigs at the age of D28 D60, D90 and D150. The abundance of Clostridium and Turicibacter was positively correlated with apparent EE digestibility. The abundance of Anaerofustis and Robinsoniella in sow fecal samples was positively correlated with apparent CF digestibility. The abundance of Collinsella and Sutterella was positively correlated with apparent NDF digestibility. The abundance of Clostridium, Collinsella, Robinsoniella and Turicibacter was positively correlated with apparent ADF digestibility. Sows have their unique gut microbial structure compared with grower pigs and some of them participate in the digestive process of different nutrients.

Increased Proportion of Fiber-Degrading Microbes and Enhanced Cecum Development Jointly Promote Host To Digest Appropriate High-Fiber Diets.

Previous study found that appropriate high-fiber diet (containing 19.10% total dietary fiber [TDF], treatment II) did not reduce apparent fiber digestibility of Chinese Suhuai finishing pigs and increased the yield of short-chain fatty acids (SCFAs), but too high-fiber diet (containing 24.11% TDF, treatment IV) significantly reduced apparent fiber digestibility compared with normal diet (containing 16.70% TDF, control group). However, characteristics of microbiota at the species level and histological structure in pigs with the ability to digest appropriate high-fiber diets were still unknown. This study conducted comparative analysis of cecal physiology and microbial populations colonizing cecal mucosa. The results showed intestinal development indexes including cecum length, densities of cecal goblet cells, and renewal of cecal epithelial cells in treatment II and IV had better performance than those in the control. Paludibacter jiangxiensis, Coprobacter fastidiosus, Bacteroides coprocola CAG:162, Bacteroides barnesiae, and Parabacteroides merdae enriched in treatment II expressed large number of glycoside hydrolase (GH)-encoding genes and had the largest number of GH families. In addition, pathogenic bacteria (Shigella sonnei, Mannheimia haemolytica, and Helicobacter felis) were enriched in treatment IV. Correlation analysis revealed that the intestinal development index positively correlated with the relative abundance of cecal mucosal microbiota and the amount of digested fiber. These results indicated that increased proportions of fiber-degrading microbes and enhanced intestinal development jointly promote the host to digest an appropriate high-fiber diet. However, although too-high fiber levels in diet could maintain the adaptive development of cecal epithelium, the proportions of pathogenic bacteria increased, which might lead to a decrease of fiber digestion in pigs. IMPORTANCE Although studies about the effects of dietary fiber on fiber digestion and intestinal microbiota of pigs were widely in progress, few studies have been conducted on the dynamic response of intestinal microbiota to dietary fiber levels, and the characteristics of intestinal microbiota and intestinal epithelial development adapted to high-fiber diet s were still unclear. Appropriate high fiber promoted the thickness of large intestine wall, increased the density of cecal goblet cells, and promoted the renewal of cecal epithelial cells. In addition, appropriate high fiber improves the microbial abundance with fiber-digesting potential. However, excessive dietary fiber caused an increase in the abundance of pathogenic bacteria. These results indicated that an increased proportion of fiber-degrading microbes and enhanced intestinal development jointly promote host to digest appropriate high-fiber diets. However, although too-high fiber levels in diet could maintain the adaptive development of cecal epithelium, the proportions of pathogenic bacteria increased, which might lead to a decrease of fiber digestion in pigs. Our data provided a theoretical basis for rational and efficient utilization of unconventional feed resources in pig production.

Effects of Varying Levels of Wheat Bran Dietary Fiber on Growth Performance, Fiber Digestibility and Gut Microbiota in Erhualian and Large White Pigs.

To evaluate the tolerance of a high-fiber diet in Erhualian pigs (Er-HL), the present investigation systematically investigated the ramifications of varying wheat bran fiber levels, specified as total dietary fiber (TDF) values of 14.07%, 16.32%, 17.99%, and 18.85%, on growth performance, fiber digestibility and gut microbiota in Er-HL, large Large White pigs (L-LW, the same physiological stage as the Er-HL) and small Large White pigs (S-LW, the same body weight as the Er-HL). Our results revealed that fiber levels exerted no discernable impact on growth performance (average daily feed intake (ADFI), and average daily gain (ADG)) of Er-HL (p > 0.05). Conversely, L-LW exhibited a decrease in ADFI and ADG with increasing fiber levels (p < 0.05). Notably, the apparent total tract digestibility (ATTD) of various fiber components, including neutral detergent fiber (NDF), acid detergent fiber (ADF), hemicellulose, TDF and insoluble dietary fiber (IDF), in Er-HL were significantly higher than those in S-LW and L-LW irrespective of diets (p < 0.05). The ATTD of cellulose and hemicellulose in Er-HL significantly decreased with increasing fiber levels (p < 0.05), yet remained statistically indifferent when comparing the 7%-wheat-bran-replaced diet (7% WRB, TDF 16.32%) to the basal diet (TDF 14.07%) (p > 0.05). The cecal microbiota of Er-HL had higher richness estimators (Chao1 and ACE) than those of S-LW and L-LW irrespective of diets (p < 0.01). Breed serves as a pivotal determinant in shaping swine gut microbiota. Thirteen genera were selected as the key bacteria related to high fiber digestibility of Er-HL. Further functional examination of these key genera elucidated an enrichment of pathways pertinent to carbohydrate metabolism in Er-HL samples compared with S-LW and L-LW samples. In summary, Er-HL exhibited high-fiber tolerance both in terms of growth performance and fiber digestibility compared with Large White pigs. Specifically, the ATTD of NDF, ADF, hemicellulose, IDF and TDF were significantly higher in Er-HL compared with L-LW and S-LW, irrespective of diets. Fiber level exerted no discernable impact on growth performance (ADFI, ADG) and the ATTD of fiber (NDF, ADF, IDF and TDF) in Er-HL. The optimum fiber level of the Er-HL was identified as 7% WRB (TDF 16.32%). Thirteen genera were ascertained to significantly contribute to high fiber digestibility of Er-HL, correlating with an enhancement of carbohydrate metabolism pathways.

Effects of short-term feeding with high fiber diets on growth, utilization of dietary fiber, and microbiota in pigs.

Finishing pigs can adapt to high-fiber diet smoothly according to the production performance and their intestinal microbiota through a 28-day trial or longer. However, it is unclear, at which stage during the experimental period, the adaptation occurred. Here we studied the dosage effects of dietary fiber (Total dietary fiber (TDF) from 16.70 to 24.11%) on growth performance, fiber digestibility, fecal microbiota, and microbial fermentation of finishing pigs during a 14-day feeding period. The results showed that the average daily feed intake (ADFI) and feed/gain (F/G) of pigs were not affected as the dietary fiber increased. Apparent total tract digestibility (ATTD) of cellulose, hemicellulose, insoluble dietary fiber (IDF), soluble dietary fiber (SDF), and TDF of pigs remained unchanged when TDF was between 16.70 and 17.75%, while strikingly decreased when TDF increased from 17.75 to 24.11%. It is worth noting that increasing fiber intake seemed to favor hemicellulose digestion. In addition, the increase in fiber intake increased fecal microbial diversity, especially improved the proportion of the members of the family Prevotellaceae, Ruminococcaceae, and Lachnospiraceae, and decreased the abundance of the genus Streptococcus. Moreover, the increase in fiber intake promoted the digestion of fiber, production of short chain fatty acids (SCFAs), and enhanced microbial pyruvate metabolism and butanoate metabolism. In conclusion, short-term high fiber feeding has no adverse effects on the growth performance of finishing pigs. ATTD of dietary fiber of finishing pigs was maintained when TDF was at 17.75%. And short-term high fiber feeding improved microbial diversity and fiber degradation functions of finishing pigs.

Metagenomic Analysis Reveals New Microbiota Related to Fiber Digestion in Pigs.

Making full use of high fiber and low-cost crop coproducts is helpful to alleviate the situation of people and livestock competing for crops. Digestion of dietary fibers in pigs is mainly through microbial fermentation in the large intestine. To reveal microbiota related to fiber digestion in pigs, fecal samples have been collected from 274 healthy female Suhuai pigs at 160 days of age under the same feeding conditions and have measured apparent neutral detergent fiber (NDF) and acid detergent fiber (ADF) digestibility. Samples from Suhuai pigs with extreme high and low apparent NDF digestibility and extreme high and low apparent ADF digestibility were subjected to shotgun metagenomic sequencing. At the species level, 62 microbial species in H_NDF group and 54 microbial species in H_ADF group were related to high fiber digestibility. Among them, Lachnospiraceae bacterium 3-1 and Alistipes sp. CAG:514 may be new types of microorganisms associated with fiber digestion. In addition, we found that more abundant GH5 and GH48 family (contribute to cellulose degradation) genes, GH39 and GH53 family (contribute to hemicellulose degradation) genes in microorganisms may contribute to the higher apparent NDF digestibility of pigs, and more abundant GH3 and GH9 family (contribute to cellulose degradation) genes in microorganisms may contribute to the higher apparent ADF digestibility of pigs. The abundance of AA4 family (helps in lignin degradation) genes in H_NDF and H_ADF groups was significantly higher than that in L_NDF and L_ADF groups, respectively (P < 0.05). Three pathways in H_NDF group and four pathways in H_ADF group are important pathways associated with degradation of non-starch polysaccharides, and their relative abundance is significantly higher than that in L_NDF and L_ADF groups, respectively. Gut microbiota of Suhuai pigs with high apparent fiber digestibility had higher abundance of genes and microbiota related to fiber digestion and may have stronger fiber digestion potential compared with low apparent fiber digestibility group. This study revealed that the characteristics of gut microbiota and microbial gene functions of pigs with high fiber apparent digestibility, which provided a theoretical basis and reference for further understanding the impact of gut microbiota on fiber digestibility of pigs.

Defatted Rice Bran Supplementation in Diets of Finishing Pigs: Effects on Physiological, Intestinal Barrier, and Oxidative Stress Parameters.

Rice bran is a waste product with low cost and high fiber content, giving it an added advantage over corn and soybean meal, which have to be purchased and always at a relatively higher cost. Under the background of increased attention to sustainable agriculture, it is significant to find alternative uses for this byproduct. A total of 35 finishing pigs were allotted to five dietary treatments: a control group with basal diet and four experimental diets where corn was equivalently substituted by 7%, 14%, 21%, and 28% defatted rice bran (DFRB), respectively. With increasing levels of DFRB, the neutrophil to lymphocyte ratio (NLR) linearly decreased (p < 0.05). In the jejunum, the mRNA level of nuclear factor erythroid-2 related factor-2 (Nrf2) exhibited a quadratic response (p < 0.01) with incremental levels of DFRB. In the colon, the mRNA levels of mucin 2 (MUC2), Nrf2, and NAD(P)H: quinone oxidoreductase 1 (NQO1) were upregulated (linear, p < 0.05) and heme oxygenase-1 (HO-1) was upregulated (linear, p < 0.01). Overall, using DFRB to replace corn decreased the inflammatory biomarkers of serum and showed potential function in modulating the intestinal barrier by upregulating the mRNA expression levels of MUC2 and downregulating that of Nrf2, NQO1, and HO-1 in the colon.

Adding Appropriate Fiber in Diet Increases Diversity and Metabolic Capacity of Distal Gut Microbiota Without Altering Fiber Digestibility and Growth Rate of Finishing Pig.

The digestion ability of pigs to dietary fiber is derived from their intestinal microbiota, especially hindgut microbiota. However, tolerance of pigs to high dietary fiber and the changes of microbiota profile with fiber levels are still unclear. To investigate the changes of gut microbiota with dietary fiber and its relationship with fiber digestibility, we conducted comparative analyses of growth rate, apparent fiber digestibility, gut microbiota and volatile fatty acid (VFA) profiles in Chinese Suhuai pigs feeding diets with different defatted rice bran (DFRB) fiber levels. We found that dietary fiber level had no effect on the growth rate of Suhuai pigs. Although the apparent digestibility of Cellulose, insoluble dietary fiber (IDF) and total dietary fiber (TDF) decreased with dietary fiber level, we found that the apparent digestibility of Cellulose, IDF and TDF of Suhuai pigs was not changed when provided with diet containing 19.10% TDF (as feed basis). The pigs provided with diet containing 19.10% TDF had higher microbial richness, proportions of several fiber-degrading bacteria taxa at genus level and predicted microbial functions (such as carbohydrate metabolism, energy metabolism) in cecum compared to those fed with basal diet. In addition, the fiber-induced increasing of fiber-degrading bacteria promoted the VFAs metabolism, which potentially helped Suhuai pigs to maintain growth rate. However, as TDF reached to 24.11% (as feed basis), the apparent digestibility of fiber decreased and the positive effect on intestine microbiota in caecum were absent. Together, our data suggest that appropriate fiber level could increase the diversity and metabolic capacity of distal gut microbiota to improve the utilization efficiency of fiber resources without altering the growth rate of pigs.

Effects of various levels of dietary fiber on carcass traits, meat quality and myosin heavy chain I, IIa, IIx and IIb expression in muscles in Erhualian and Large White pigs.

This study evaluated the effects of bran fiber levels on carcass traits, meat quality and expression of myosin heavy chain isoform genes in muscles in Erhualian (Er-HL) and Large White pig (LW). Our results showed that fiber level did not affect carcass weight of Er-HL, while carcass weight of LW decreased with the increase of fiber level. Fiber level did not influence meat quality traits of LW, whereas increased fiber level led to an increase in pH45 min of Er-HL, and Er-HL fed 7% bran fiber (BRC) increased redness of meat compared with control diet (CON). We observed a decreased MyHCIIb and MyHCIIx mRNAs and protein levels in Er-HL fed 7% BRC compared with those in other groups, accompanying with a tendency for increased mRNA abundance of MyHCI. The slow-twitch oxidative fiber (MyHCI) is rich in myoglobin and mitochondrial oxidative metabolic enzymes, and the increased expression of MyHCI fiber in pork has resulted in higher redness value. In summary, bran fiber had no adverse effect on the meat quality of LW and appropriate fiber level addition in diets could improve meat quality of Er-HL through regulation the expression of myofiber types.

Effects of Increasing Levels of Defatted Rice Bran on Intestinal Physical Barrier and Bacteria in Finishing Pigs.

The aims of this study were to assess the effects of increasing levels of DFRB as a replacement for corns on intestinal physical barrier function and bacteria of finishing pigs. A total of 35 castrated finishing pigs (age: 158.5 ± 2.0 d, initial body weight: 62.9 ± 0.8 kg) were randomly divided into five dietary treatments (seven replicates/treatment) for a 28-day experimental period, i.e., a control diet with basal diet, and four experimental diets in which maize was replaced by 7%, 14%, 21%, and 28% DFRB, respectively. The results showed that serum endotoxins concentration and diamine oxidase (DAO) activity were both increased (linear, p = 0.0004, 0.001, respectively) with DFRB level. However, compared with control group, serum endotoxins concentration and DAO activity were not different in pigs fed with 7% DFRB in the diet. There was a quadratic response in serum D-lactate concentration to the increased DFRB (quadratic, p = 0.021). In the cecum, thickness of the intestinal wall significantly increased with increasing levels of DFRB in the diets (linear, p = 0.033), while crypt depth/thickness of the intestinal wall ratio significantly decreased with increasing level of DFRB in the diets (linear, p = 0.043). In the jejunum, total bacteria, Escherichia coli, and Bifidobacterium all responded quadratically to increasing levels of DFRB in the diets (quadratic, p = 0.003, 0.001, 0.006, respectively). Additionally, there was no difference in Escherichia coli in pigs fed 0%, 7%, and 14% DFRB diets. In the colon, there were quadratic responses in C. perfringens to the increased DFRB (quadratic, p = 0.023). C. perfringens reduced as the DFRB concentration increased from 0% to 14% and then increased. When D-lactate, total bacteria, Escherichia coli, Bifidobacterium, and C. perfringens were considered, the optimal substitution level of DFRB were 12.00%, 11.84%, 7.50%, 8.92%, and 15.92%, respectively. In conclusion, 7% DFRB had a beneficial effect on intestinal wall thickness, Bifidobacterium and C. perfringens, and had no adverse effect on intestinal permeability and Escherichia coli.