Effects of Enterotoxigenic Escherichia coli Challenge on Jejunal Morphology and Microbial Community Profiles in Weaned Crossbred Piglets.

Pathogenic enterotoxigenic Escherichia coli (ETEC) is a major cause of bacterial diarrhea in weaning piglets, which are vulnerable to changes in environment and feed. This study aimed to determine the effects of the ETEC challenge on piglet growth performance, diarrhea rate, jejunal microbial profile, jejunal morphology and goblet cell distribution. A total of 13 piglets from one litter were selected on postnatal day 21 and assigned to treatments with or without ETEC challenge at 1 × 108 CFUs, as ETEC group or control group, respectively. On postnatal day 28, samples were collected, followed by the detection of serum biochemical indexes and inflammatory indicators, HE staining, PAS staining and 16S rDNA gene amplicon sequencing. Results showed that the growth performance decreased, while the diarrhea rate increased for the ETEC group. The jejunum is the main segment of the injured intestine during the ETEC challenge. Compared with the control, the ETEC group displayed fewer goblet cells in the jejunum, where goblet cells are more distributed at the crypt and less distributed at the villus. In addition, ETEC piglets possessed higher abundances of the genus Desulfovibrio, genus Oxalobacter and genus Peptococus and lower abundances of the genus Prevotella 2, genus Flavonifractor and genus Blautra. In terms of alpha diversity, Chao 1 and observed features indexes were both increased for the ETEC group. Our study provides insights into jejunal histopathological impairment and microbial variation in response to ETEC infection for weaned piglets and is a valuable reference for researchers engaged in animal health research to select stress models.

Prediction of available energy and amino acid digestibility of Chinese sorghum fed to growing-finishing pigs.

Two experiments were conducted to determine digestible energy (DE), metabolizable energy (ME), as well as the standardized ileal digestibility (SID) of crude protein (CP) and amino acids (AA) in 10 sorghum samples fed to pigs. In experiment 1, 22 crossbred barrows (Duroc × Yorkshire × Landrace, Initial body weight [BW]: 70.0 ±â€…1.8 kg) were selected and allotted to a replicated 11 × 3 incomplete Latin square design, including a basal diet and 10 sorghum energy diets and three consecutive periods. Each period had 7 d adaptation and 5 d total feces and urine collection. The DE and ME were determined by the total collection and the difference method. In experiment 2, 22 crossbred barrows (Duroc × Yorkshire × Landrace, Initial BW: 41.3 ±â€…1.2 kg) that had a T-cannula installed in the distal ileum were assigned to a replicated 11 × 3 incomplete Latin square design, including an N-free diet and 10 sorghum diets. Each period had 5 d adaptation and 2 d ileal digesta collection. The basal endogenous N losses were measured by the N-free diet method. All diets in experiment 2 were added 0.30% titanium dioxide as an indigestible marker for calculating the ileal CP and AA digestibility. On an as-fed basis, the DE and ME contents in sorghum were 3,410 kcal/kg (2,826 to 3,794 kcal/kg) and 3,379 kcal/kg (2,785 to 3,709 kcal/kg), respectively. The best-fit prediction equation for DE and ME were DE = 6,267.945 - (1,271.154 × % tannin) - (1,109.720 × % ash) (R2 = 0.803) and ME = 51.263 + (0.976 × DE) (R2 = 0.994), respectively. The SID of CP, Lys, Met, Thr, and Trp (SIDCP, SIDLys, SIDMet, SIDThr, and SIDTrp) in 10 sorghum samples were 78.48% (69.56% to 84.23%), 74.27% (61.11% to 90.60%), 92.07% (85.16% to 95.40%), 75.46% (66.39% to 80.80%) and 87.99% (84.21% to 92.37%), respectively. The best prediction equations for SID of CP and the first four limiting AAs were as following: SIDCP = 93.404 - (21.026 × % tannin) (R2 = 0.593), SIDCP = 42.922 - (4.011 × % EE) + (151.774 × % Met) (R2 = 0.696), SIDLys = 129.947 - (670.760 × % Trp) (R2 = 0.821), SIDMet = 111.347 - (232.298 × % Trp) (R2 = 0.647), SIDThr = 55.187 + (3.851 × % ADF) (R2 = 0.609) and SIDTrp = 95.676 - (10.824 × % tannin) (R2 = 0.523), respectively. Overall, tannin and ash are the first and second predictors of DE and ME values of sorghum, respectively, and the tannin, EE, Trp, ash, CF, and ADF can be used as the key predictors for SID of CP and first four limiting AAs.

In this manuscript, we selected and analyzed the chemical composition of 10 different varieties of Chinese sorghum. Two digestion and metabolism experiments were conducted to measure the concentrations of available energy (digestible energy and metabolizable energy) and ileal amino acid digestibility (apparent or standardized ileal amino acid digestibility) in sorghums fed to pigs. The results showed as following: 1) The available energy values of 10 sorghum samples varied greatly, and the ileal digestibility of some amino acids also differed. 2) The available energy value and the ileal digestibility of crude protein and amino acids can be predicted by the contents of chemical components in sorghum, such as tannin (a substance in sorghum that can bind to biological macromolecules like proteins), ash, tryptophan, and fiber composition, etc. The above findings enrich the ingredient database and assist for feed enterprises to make precise formulations when using the Chinese sorghum.

Antioxidant potential of Pediococcus pentosaceus strains from the sow milk bacterial collection in weaned piglets.

BACKGROUND: In modern animal husbandry, breeders pay increasing attention to improving sow nutrition during pregnancy and lactation to favor the health of neonates. Sow milk is a main food source for piglets during their first three weeks of life, which is not only a rich repository of essential nutrients and a broad range of bioactive compounds, but also an indispensable source of commensal bacteria. Maternal milk microorganisms are important sources of commensal bacteria for the neonatal gut. Bacteria from maternal milk may confer a health benefit on the host. METHODS: Sow milk bacteria were isolated using culturomics followed by identification using 16S rRNA gene sequencing. To screen isolates for potential probiotic activity, the functional evaluation was conducted to assess their antagonistic activity against pathogens in vitro and evaluate their resistance against oxidative stress in damaged Drosophila induced by paraquat. In a piglet feeding trial, a total of 54 newborn suckling piglets were chosen from nine sows and randomly assigned to three treatments with different concentrations of a candidate strain. Multiple approaches were carried out to verify its antioxidant function including western blotting, enzyme activity analysis, metabolomics and 16S rRNA gene amplicon sequencing. RESULTS: The 1240 isolates were screened out from the sow milk microbiota and grouped into 271 bacterial taxa based on a nonredundant set of 16S rRNA gene sequencing. Among 80 Pediococcus isolates, a new Pediococcus pentosaceus strain (SMM914) showed the best performance in inhibition ability against swine pathogens and in a Drosophila model challenged by paraquat. Pretreatment of piglets with SMM914 induced the Nrf2-Keap1 antioxidant signaling pathway and greatly affected the pathways of amino acid metabolism and lipid metabolism in plasma. In the colon, the relative abundance of Lactobacillus was significantly increased in the high dose SMM914 group compared with the control group. CONCLUSION: P. pentosaceus SMM914 is a promising probiotic conferring antioxidant capacity by activating the Nrf2-Keap1 antioxidant signaling pathway in piglets. Our study provided useful resources for better understanding the relationships between the maternal microbiota and offspring. Video Abstract.

Comparisons of carcass traits, meat quality, and serum metabolome between Shaziling and Yorkshire pigs.

This study aims to compare the meat quality of Shaziling and Yorkshire pigs and to find the potential indicator in serum for superior meat quality. Six Shaziling and Yorkshire pigs at 30, 60, 90, 150, 210, and 300 d of age were selected to examine carcass traits, meat quality, and serum metabolome. The results showed that the body weight, carcass length, and loin eye area of Shaziling pigs at 150, 210, and 300 d of age were significantly lower than those of Yorkshire pigs (P < 0.05). Shaziling pigs at 150 and 300 d of age had significantly lower backfat thickness than Yorkshire pigs (P < 0.05). Compared with Yorkshire pigs, Shaziling pigs at all 6 ages had a lower lean percentage and a higher fat percentage (P < 0.05). At 60, 90, and 150 d of age, the post-mortem pH-decline, b∗ value (yellowness), and drip loss of Shaziling pigs were significantly lower than those of Yorkshire pigs (P < 0.05). Moreover, at 150 d of age, Shaziling pigs had significantly higher a∗ value (redness) and intramuscular fat (IMF) content than Yorkshire pigs (P < 0.05). Correlation analysis between the top 40 metabolites and phenotypes indicated that L-carnitine had positive correlations with fat percentage, pH24h, and IMF content, but had negative correlations with lean percentage, L∗ value (lightness), and b∗ value (P < 0.05). Serum L-carnitine content, fat percentage, pH24h, and IMF content all decreased first and then increased as the pigs grew, which verified the positive correlations between L-carnitine and these phenotypes. In conclusion, Shaziling pigs have a slower growth rate but a better meat quality than Yorkshire pigs. The meat quality of Shaziling pigs is the best from 150 to 210 d of age. This study suggests that a higher serum L-carnitine content is a promising indicator for better meat quality.

Mitochondrial Iron Metabolism: The Crucial Actors in Diseases.

Iron is a trace element necessary for cell growth, development, and cellular homeostasis, but insufficient or excessive level of iron is toxic. Intracellularly, sufficient amounts of iron are required for mitochondria (the center of iron utilization) to maintain their normal physiologic function. Iron deficiency impairs mitochondrial metabolism and respiratory activity, while mitochondrial iron overload promotes ROS production during mitochondrial electron transport, thus promoting potential disease development. This review provides an overview of iron homeostasis, mitochondrial iron metabolism, and how mitochondrial iron imbalances-induced mitochondrial dysfunction contribute to diseases.

Postnatal growth retardation is associated with intestinal mucosa mitochondrial dysfunction and aberrant energy status in piglets.

Individuals with postnatal growth retardation (PGR) are prone to developing chronic disease. Abnormal development in small intestine is casually implicated in impaired growth performance. However, the exact mechanism is still unknown. In this present study, PGR piglets (aged 42 days) were employed as a good model to analyse changes in nutrient absorption and energy metabolism in the intestinal mucosa. The results showed lower serum concentrations of free amino acids, and lipid metabolites in PGR piglets, which were in accordance with the down-regulated mRNA expressions involved in fatty acid and amino acid transporters in the jejunal and ileal mucosa. The decreased activities of digestive enzymes and the marked swelling in mitochondria were also observed in the PGR piglets. In addition, it was found that lower ATP production, higher AMP/ATP ratio, deteriorated mitochondrial complex III and ATP synthase, and decreased manganese superoxide dismutase activity in the intestinal mucosa of PGR piglets. Furthermore, altered gene expression involved in energy metabolism, accompanied by decreases in the protein abundance of SIRT1, PGC-1α and PPARγ, as well as phosphorylations of AMPKα, mTOR, P70S6K and 4E-BP1 were observed in intestinal mucosa of PGR piglets. In conclusion, decreased capability of nutrient absorption, mitochondrial dysfunction, and aberrant energy status in the jejunal and ileal mucosa may contribute to PGR piglets.

Hepatic lipid metabolism is affected by a daily 3-meal pattern with varying dietary crude protein with a pig model.

The present study was conducted to evaluate the effects of 3 meals administered daily with varying dietary crude protein (CP) contents on hepatic lipid metabolism with a pig model. Pigs were divided into 3 groups according to the following feeding patterns: feeding a basal CP diet 3 times daily (3C); feeding a high CP diet for breakfast, the basal CP diet for lunch, and a low CP diet for dinner (HCL); and feeding the low CP diet for breakfast, the basal CP diet for lunch, and the high protein diet for dinner (LCH). Three groups took equivalent diet per meal ensuring that every pig was fed with similar dietary formulae daily. Results showed that HCL feeding pattern reduced the relative kidney weight (P < 0.05), and LCH feeding pattern increased the relative liver weight of pigs (P < 0.05) when compared with those in the 3C group. Plasma urea nitrogen (P < 0.01) and lipase (P < 0.05) decreased in the HCL group but increased in the LCH group. Both HCL and LCH feeding patterns reduced plasma triglycerides (P < 0.01), non-esterified fatty acids (NEFA) (P < 0.01), and hepatic crude fat (0.05 < P < 0.10) of pigs. Real-time quantitative PCR (RT-qPCR) results showed that dynamic feeding patterns down-regulated (P < 0.05) the mRNA level of lipid metabolism related genes, including adipose triglyceride lipase (ATGL), acetyl-CoA carboxylase (ACCα), liver X receptor (LXRα) in the liver, and negatively regulate elements of circadian clock, including period 1 (Per1), period 2 (Per2), cryptochrome (Cry2), which in turn, upregulated (P < 0.05) the protein expression of positive regulate element brain and muscle Arnt-like 1 (BMAL1) when compared with 3C group. Overall, our findings suggested that dynamic feeding patterns may affect hepatic lipid metabolism via regulation of the circadian clock.

ß-Hydroxy-ß-methylbutyrate modulates lipid metabolism in adipose tissues of growing pigs.

BACKGROUND: The effects and roles of the leucine (Leu) metabolite ß-hydroxy-ß-methylbutyrate (HMB) in lipid metabolism in adipose tissues of pigs are still unknown. OBJECTIVES: This study was conducted to investigate the effects of excess Leu versus HMB on growth, carcass traits, and lipid metabolism in adipose tissues of growing pigs. METHODS AND RESULTS: Compared to control, the Leu/HMB group significantly increased/reduced weight of total fat mass, respectively, with a concurrent increase of serum adiponectin concentration (P < 0.05). Moreover, dietary HMB supplementation regulated the expression of genes involved in adipose tissue function, accompanied by increases/decreases in the phosphorylation of AMPKα/mTOR in perirenal adipose tissue, respectively (P < 0.05). Serum IL-15 concentration and the mRNA abundance of IL-15, PGC-1α, and NRF-1 were also increased in the HMB group (P < 0.05). CONCLUSIONS: HMB supplementation can regulate adipose tissue function including fatty acid oxidation, lipolysis, and adipokine secretion. These effects may be partly mediated by AMPKα-mTOR pathway and associated with mitochondrial biogenesis, the AMPK-PGC-1α axis, and myokines secreted by muscle tissues.

Effects of a daily three-meal pattern with different dietary protein contents on pig growth performance, carcass and muscle quality traits.

BACKGROUND: Recent research related to phase-feeding programmes for pig nutrition do not always account for the variation among individuals, and feeds are usually formulated to optimise the performance of the whole pig population. This study aimed at measuring the effects of a daily three-meal pattern with different dietary protein contents on pig growth performance, carcass and muscle quality traits. RESULTS: The results showed that compared with the 3C treatment, average daily gain (ADG) of pigs in the HCL treatment increased by 14.75% (P < 0.05) during period 1. The carcass weight (P = 0.006) and slaughter weight (P = 0.021) in the HCL group increased when compared with those in the 3C and LCH treatments. Moreover, the LCH feeding sequences contributed to reduce the drip loss in longissimus dorsi (LD) muscle (P < 0.05) when compared with the 3C treatment. The HCL or LCH feeding sequence contributed to increase the meat quality when compared with those receiving the 3C treatment. CONCLUSION: Collectively, our results indicate that feeding high protein meal in the morning and a gradual reduction of the protein content in meals over the day may improve muscle quality characteristics, maximise performance, and reduce the pig feed cost. © 2017 Society of Chemical Industry.

A Maternal Two-meal Feeding Sequence with Varying Crude Protein Affects Milk Lipid Profile in A Sow-Piglet Model.

The effects of a two-meal feeding sequence on production performance and milk lipid profile were investigated. Sixty pregnant sows (d 85 of gestation) were assigned to 3 groups: 2 C group (fed a control crude protein [CP] diet at 0600 and 1500 daily), LH group (fed a low CP diet and a high CP diet at 0600 and 1500), or HL group (fed a high CP diet and a low CP diet at 0600 and 1500). Reproductive performance of sows, and lipid profiles of plasma and milk were measured. Results showed that the HL feeding sequence dramatically increased average piglet weight/litter, average daily gain of piglet/litter, and milk production of sows. LH feeding sequence increased milk fat proportion, and HL feeding sequence significantly increased the proportion of milk MUFA on d 14 and 21 of lactation. Interestingly, the HL feeding sequence also reduced the ratio of C18:1 cis /C18:1 trans in milk, which may account for the greater milk production of sows and growth performance of piglets during lactation. These findings indicated that both the maternal two-meal feeding sequences with varying crude protein improved milk production and milk lipid profiles of sows, which might contribute to improving growth performance of piglets.

Diurnal variations in polyunsaturated fatty acid contents and expression of genes involved in their de novo synthesis in pigs.

The daily variations in circulating fatty acid (FA) contents and lipid metabolism have been well documented. However, whether long chain polyunsaturated FA (PUFA) contents and expression of genes involved in their de novo synthesis exhibit daily rhythms are yet unknown. We conducted the present study to investigate the daily variations in PUFA contents in plasma and liver of pigs. Moreover, diurnal expression of genes encode fatty acid desaturases and elongases, which are key enzymes catalyzed de novo synthesis of long chain PUFA, were also explored. The results showed that long chain PUFA contents in plasma and liver both exhibited diurnal rhythms. Diurnal variations were also observed in mRNA expression of FASD1 (Delta 5-desaturase), FASD2 (Delta 6-desaturase), ELOVL5 (fatty acid elongase 5) and ELOVL2 in liver, with an unexpectedly high level at night. Moreover, our results showed a similarity between the diurnal patterns of FASD1, FASD2, ELOVL2, ELOVL5 and Period 2. These results indicated a high activity of the desaturase-elongase pathway at night in pigs. These findings have important physiological and pathophysiological implications, since long chain PUFA are essential for cell function and closely involved in the development of metabolic syndrome.

Chitosan oligosaccharide affects antioxidant defense capacity and placental amino acids transport of sows.

BACKGROUND: Chitosan oligosaccharide (COS) is widely consumed as a functional food due to its multiple health effects, but few studies about COS supplement on placental antioxidant and nutrition transport capacity were reported. Taken pregnant sow as a model, we aimed to investigate the effects of dietary COS supplementation during late gestation on placental amino acids transport and antioxidant defense capacity of sows. From day (d) 85 of gestation to parturition, sixteen pregnant sows were divided into a control group (basal diet without COS supplementation) and a COS group (30 mg COS/kg basal diet). Plasma sample of sow was collected on d 110 of gestation, and placenta tissue was obtained during parturition. Then plasma antioxidant enzyme's activities, the relative level of oxidant stress related genes, amino acids transport related genes and mTOR pathway molecules in placenta were determined. RESULTS: Results showed that maternal dietary supplementation with COS increased (P < 0.05) plasma total SOD, caused a downtrend in plasma MDA (0.05 < P < 0.10) on d 110 of gestation. Interestingly, the mRNA expression of some antioxidant genes in the placenta were increased (P < 0.05) and pro-inflammatory cytokines were reduced (P < 0.05) by COS supplement, whereas no significant difference was observed in the activities of placental total SOD and CAT between two groups. Additionally, further study demonstrated that COS feeding stimulated mTOR signaling pathway, increased amino acids transporters expression in placenta. CONCLUSIONS: These observations suggested that COS supplement in sow's diet during late gestation enhanced antioxidant defense capacity of sows, promoted placental amino acids transport, which may contribute to the health of sows and development of fetus during gestation.

Dietary supplementation of Lonicera macranthoides leaf powder improves amino acid profiles in serum and longissimus thoracis muscle of growing-finishing pigs.

A 60-days feeding trial was conducted to determine the effect of dietary supplementation of Lonicera macranthoides leaf (LML) on growth performance, meat quality, and free amino acid profiles in growing-finishing pigs. L. macranthoides leaves were harvested, air-dried and milled to powder. A total of 288 crossbred barrows (Large White × Landrace) with an average initial BW of 54.0 ± 3.4 kg were randomly allocated to 3 treatment groups and fed either a basal diet or a diet supplemented with 0.5% or 1.0% LML powder. Results showed that diet supplemented with LML powder did not affect average daily feed intake (ADFI) and feed: gain ratio (P > 0.05) in growing-finishing pigs. Moreover, diet supplemented with LML powder showed no significant effects on carcass traits (P > 0.05) including dressing percentage, backfat thickness and loin muscle area, and meat quality traits (P > 0.05) including meat color, intramuscular fat and cooking loss. However, diet supplemented with LML powder significantly improved free amino acid profiles in serum and longissimus thoracis muscle in growing-finishing pigs. Most importantly, dietary LML powder increased total free amino acids content (P < 0.001) and essential free amino acid content (P < 0.001) in longissimus thoracis muscle. These results indicate that LML has the potential to improve the nutritional value of meat through improving free amino acid profiles.

Effects of the Sequence of Isocaloric Meals with Different Protein Contents on Plasma Biochemical Indexes in Pigs.

Nutrient composition and pattern of food intake may play a significant role in weight gain. The aim of this study was to document the effects of a daily 3-meal pattern with isocaloric diets containing different dietary protein contents on growth performance and different plasma biochemical indexes including amino acid plasma concentration in castrated male pigs. Then, 21 DLY (Duroc×Landrace×Yorkshire) pigs aged 60 days were assigned randomly into 3 groups: a control group (crude protein, CP 18.1%), a group receiving high then basal and then low CP meals (High-Low group) and a group receiving low then basal and then high CP meal (Low-High group) for 40 days with pigs being feed-restricted. On day 40, after 12 h fasting, blood samples were obtained for analysis. The results showed that the insulin/glucagon ratio was lower in the High-Low group (P<0.05) when compared with the control group. Compared with the control group, the average daily gain of pigs from the High-Low group increased by 14.10% (P = 0.046). Compared with the control group, serum gamma-glutamyl transferase (GGT) decreased significantly (P<0.05) in both the High-Low and Low-High groups. Plasma concentrations of branched-chain amino acids (BCAA: valine, isoleucine and leucine) increased in the Low-High group (P<0.05) when compared with the control group; and plasma methionine and serine decreased in both the two experimental groups (P<0.05). Compared with the High-Low group, all the BCAA increased significantly (P<0.05) in the Low-High group. These findings suggest that the sequence and quantity of alimentary protein intake affect the insulin/glucagon ratio, as well as amino acid concentrations including BCAA, methionine and serine. It is proposed that meal pattern with pigs receiving high then basal and then low CP meals daily may help to improve the weight gain of pigs.

Supplementation of the sow diet with chitosan oligosaccharide during late gestation and lactation affects hepatic gluconeogenesis of suckling piglets.

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<0.05), and the concentration of amino acids in sow milk (P<0.05). Meanwhile, maternal supplementation with COS increased (P<0.05) mRNA expression levels and activities of PEPCK-C, PEPCK-M and G6Pase in the liver of piglets compared with piglets from control fed sows. Correspondingly, the level of plasma glucose was higher (P<0.001) and hepatic glycogen was lower (P<0.05) in piglets from COS fed sows when compared with that in the control group. In conclusion, dietary supplementation of the diet with COS during late gestation and lactation reduced piglet hypoglycemia by stimulating hepatic gluconeogenesis and improved the growth rate of suckling piglets.