Supplemental Clostridium butyricum modulates lipid metabolism by reshaping the gut microbiota composition and bile acid profile in IUGR suckling piglets

Background Intrauterine growth restriction(IUGR)can cause lipid disorders in infants and have long-term adverse effects on their growth and development.Clostridium butyricum(C.butyricum),a kind of emerging probiotics,has been reported to effectively attenuate lipid metabolism dysfunctions.Therefore,the objective of this study was to investigate the effects of C.butyricum supplementation on hepatic lipid disorders in IUGR suckling piglets.Methods Sixteen IUGR and eight normal birth weight(NBW)neonatal male piglets were used in this study.From d 3to d 24,in addition to drinking milk,the eight NBW piglets(NBW-CON group,n=8)and eight IUGR piglets(IUGR-CON group,n=8)were given 10 mL sterile saline once a day,while the remaining IUGR piglets(IUGR-CB group,n=8)were orally administered C.butyricum at a dose of 2×108colony-forming units(CFU)/kg body weight(suspended in 10 mL sterile saline)at the same frequency.Results The IUGR-CON piglets exhibited restricted growth,impaired hepatic morphology,disordered lipid metabolism,increased abundance of opportunistic pathogens and altered ileum and liver bile acid(BA)profiles.However,C.butyricum supplementation reshaped the gut microbiota of the IUGR-CB piglets,characterized by a decreased abundance of opportunistic pathogens in the ileum,including Streptococcus and Enterococcus.The decrease in these bile salt hydrolase(BSH)-producing microbes increased the content of conjugated BAs,which could be transported to the liver and function as signaling molecules to activate liver X receptorα(LXRα)and farnesoid X receptor(FXR).This activation effectively accelerated the synthesis and oxidation of fatty acids and down-regulated the total cholesterol level by decreasing the synthesis and promoting the efflux of cholesterol.As a result,the growth performance and morphological structure of the liver improved in the IUGR piglets.Conclusion These results indicate that C.butyricum supplementation in IUGR suckling piglets could decrease the abundance of BSH-producing microbes(Streptococcus and Enterococcus).This decrease altered the ileum and liver BA profiles and consequently activated the expression of hepatic LXRαand FXR.The activation of these two signaling molecules could effectively normalize the lipid metabolism and improve the growth performance of IUGR suckling piglets.

Supplementation with alpha-glycerol monolaurate during late gestation and lactation enhances sow performance,ameliorates milk composition,and improves growth of suckling piglets

Background Physiological changes during lactation cause oxidative stress in sows,reduce immunity,and hamper the growth capacity of piglets.Alpha-glycerol monolaurate(α-GML)has potential for enhancing the antimicrobial activity of sows and the growth of suckling piglets.Methods Eighty sows were allocated randomly to four groups:basal diet and basal diets supplemented with 500,1000,or 2000 mg/kgα-GML.The experiment started on d 85 of gestation and lasted until piglets were weaned on d 21 of lactation.The number of live-born piglets was standardized to 12 tion,body weight of piglets was measured and milk samples±1 per sow on day of parturition.On d 0 and21 of lactawere obtained from sows,and serum samples and feces from piglets were obtained on d 21.Results Feed intake,backfat loss,and weaning estrus interval did not differ among the four groups of sows.Maternalα-GML supplementation increased(P<0.05)the body weight of piglets at weaning and the apparent total tract digestibility of crude fat of sows.The immunoglobulin A and immunoglobulin G levels were greater(P<0.05)in a quadratic manner in the milk of sows as dietaryα-GML increased.Concerning fatty acid profile,C12:0,C15:0,C17:0,C18:2n6c,C18:3n3,C24:0,and C22:6n3 were higher(P<0.05)in linear and quadratic manners in colostrum of sowsfedα-GML diets compared with the control sows.There was lower(P<0.05)n-6:n-3 polyunsaturated fatty acid ratio in milk than in the control sows.Maternalα-GML increased the abundance of Firmicutes(P<0.05)and decreased the abundance of Proteobacteria(P<0.05)of piglet fecal microbiota.Conclusions Dietary supplementation withα-GML improved milk immunoglobulins and altered fatty acids of sows,thereby improving the health of piglets.

Dietary arginine supplementation in multiparous sows during lactation improves the weight gain of suckling piglets

This study investigated the effects of dietary arginine （Arg） supplementation, just during lactation, on sow and litter per- formance, plasma concentrations of metabolites and hormones, and milk yield and composition in multiparous sows. Thirty-one sows were randomly assigned to 3 dietary treatments supplemented with 0.0 （control, n=10）, 0.5% （n=10）, or 1.0% （n=11） L-Arg-HCI, respectively. Experimental diets were provided to the sows from d 3 to 21 of lactation. Plasma and milk samples were collected at d 14 and 21 of lactation. The average daily gain （ADG） of piglets from sows fed diets supplemented with 0.5 or 1.0% L-Arg-HCI at d 3 to 14 of lactation, were higher than that of controls （P〈0.05）. Maternal supplementation with 1.0% L-Arg-HCI also increased ADG of piglets between d 3 and 21 of lactation than that of the controls （P〈0.05）. There was no significant effect of supplementation on average daily feed intake （ADFI）, body weight loss, and backfat thickness loss of lactating sows. Supplementation with 0.5 or 1.0% L-Arg-HCI had a trend towards increasing milk yields and milk fat contents （0.05〈P〈0.10）; milk protein and lactose were unchanged. Supplementation with 1.0% L-Arg- HCI increased plasma concentrations of prolactin and insulin in sows at d 14 and 21 of lactation, and plasma concentrations of non-esterified fatty acid （NEFA）, insulin-like growth factor-1 （IGF-1）, and nitric oxide （NO） in sows at d 21 of lactation, when compared to the controls （P〈0.05）. Supplementation with 1.0% L-Arg-HCI increased IGF-1 and spermine in milk at d 14 of lactation, relative to the controls （P〈0.05）. Plasma Arg concentrations at d 14 and 21 of lactation, as well as plasma NO level and milk IGF-1 at d 21 of lactation, were increased, while plasma urea nitrogen （PUN） concentration at d 14 and 21 of lactation was decreased, by supplementation with 0.5 or 1.0% L-Arg-HCI when compared to the controls （P〈0.05）. Collectively, dietary supplementation of multiparous sows with Arg, just during lactation, is beneficial for enhancing litter weight gain but the complete mechanism remains to be determined and may involve in the maternal endocrine changes and milk polyamines contents.

Early-life galacto-oligosaccharides supplementation alleviates the small intestinal oxidative stress and dysfunction of lipopolysaccharide-challenged suckling piglets

Background:Galacto-oligosaccharides(GOS)are non-digestible food ingredients that promote the growth of beneficial bacteria in the gut.This study investigated the protective effect of the early-life GOS supplement on the piglets’gut function against the oxidative stress induced by lipopolysaccharide(LPS)-challenge.Methods:Eighteen neonatal piglets were assigned to three groups including CON,LPS and LPS+GOS groups.The piglets in CON group and LPS group received physiological saline,while those in LPS+GOS group received GOS solution for 13 d after birth.On d 14,the piglets in LPS group and LPS+GOS group were injected with LPS solutions,while the piglets in CON group were injected with the same volume of physiological saline.Results:The results showed that the early-life GOS supplement blocked the LPS-induced reactive oxygen species(ROS)secretion,malondialdehyde(MDA)production and the increase of pro-apoptotic factor expression.Meanwhile,the early-life GOS supplement improved the activities of antioxidant enzymes,disaccharidase enzymes activities,and digestive enzymes activities,and increased the mRNA abundance of the gene related to nutrient digestion and absorption and the relative protein expression of tight junction.The study also showed that the early-life GOS supplement improved the expression of Hemeoxygenase-1(HO-1)and NAD(P)H/quinone acceptor oxidoreductase-1(NQO-1),and activated the AMP-activated protein kinase(AMPK).Conclusions:These results suggested that GOS enhanced the gut function,reduced the ROS production and proapoptotic factors gene expression,and activated the AMPK signaling pathway in LPS-challenged piglets.

Synthesis and Metabolism of 5-hydroxytryptamine in Peripheral Circulatory System of Suckling Piglets

[ Objective] To find the rule of 5-hydroxytryptamine （5-HT） synthesis and metabolism in peripheral blood of suckling piglets with aging. [ Method ] All of 15 British Large White pigs at 0, 5 and 15 d of age were selected as experimental animals, and their serumal concentrations of L- tryptophan （L-Trp）, 5-HT and 5-hydroxyindoleacetic acid （5-HIAA） in peripheral circulatory system were determined synchronously by reversed- phase high-performance liquid chromatography method with UV. [ Result] The concentration of L-Trp was （16. 92 ± 1.74） iJmol/L at birth, （59.94 ±10.88） iJmol/L at 5 d of age and （70.42± 6.48） μmol/L at 15 d of age. The concentration of 5-HT was （ 12.85 ± 1.79） iJmol/L at birth, （4.81 ±2.05） ijmol/L at 5 d of age and （5.49±1.09） μmol/L at 15 d of age. While 5-HIAA, the metabolic product of 5-HT, was not detected at all the samples selected. [ Conclusion] The L-Trp concentration of suckling piglets increase significantly with aging （ P 〈0. 05）. While the concentra- tion of 5-HT and the conversion ratio of L-Trp to 5-HT are both highest at birth （P〈0.05） and keep stable at 5 and 15 d of age.

Mechanism of iron on the intestinal epithelium development in suckling piglets

This study aimed to investigate the mechanism of iron on intestinal epithelium development of suckling piglets. Compared with newborn piglets, 7-day-old and 21-day-old piglets showed changes in the morphology of the jejunum, increased proliferation,differentiated epithelial cells, and expanded enteroids. Intestinal epithelium maturation markers and iron metabolism genes were significantly changed. These results suggest that lactation is a critical stage in intestinal epithelial development, accompanied by changes in iron metabolism. In addition, deferoxamine(DFO) treatment inhibited the activity of intestinal organoids at passage 4(P4) of 0-day-old piglets, but no significant difference was observed in epithelial maturation markers at passage 1(P1) and P4,and only argininosuccinate synthetase 1(Ass1) and β-galactosidase(Gleb) were up-regulated at passage 7(P7). These results in vitro show that iron deficiency may not directly affect intestinal epithelium development through intestinal stem cells(ISCs). The iron supplementation significantly down-regulated the m RNA expression of interleukin-22 receptor subunit alpha-2(IL-22RA2)in the jejunum of piglets. Furthermore, the m RNA expression of IL-22 in 7-day-old piglets was significantly higher than that in0-day-old piglets. Adult epithelial markers were significantly up-regulated in organoids treated with recombinant murine cytokine IL-22. Thus, IL-22 may play a key role in iron-affecting intestinal epithelium development.

Supplementing oat hulls to the diet of suckling piglets altered their intestinal tract and colonic microbiota development

Current study evaluated the effect of a fine and coarsely ground insoluble dietary fibre source on the gastrointestinal development of suckling pigs.Oat hulls(OH)were selected as a model feedstuff,rich in cellulose,lignin,and insoluble dietary fibre.Three experimental supplemental diets were formulated:a finely ground,low fibre and nutrient dense diet served as control(CON).For the 2 high fibre diets,15%heat-treated starch in CON was exchanged with OH,either finely(OH-f)or coarsely ground(OH-c).Litters of 10 primi-and multiparous sows(mean litter size 14.6±0.84)were used.Within a litter,experimental diets were allotted to triplets of 4 piglets.From approximately 12 d of age,piglets’individual feed intakes were recorded 2 times per day when separated from their dam for 70 min.Piglets could suckle with their dam for the remainder of the day.On d 24 and 25,from the total pool of 120 piglets,seven healthy well-eating piglets per treatment were selected for post-mortem evaluation,resulting in 14 replicates per treatment.Consumption of OH-c and OH-f did not impede clinical health and production performance of piglets.The full stomach weights tended to be greater for OH-c compared to OH-f whereas CON was intermediate(P=0.083).Supplementing OH significantly increased ileal villus height and caecal dry matter concentration(P<0.05).For the colon,OH increased its length,contents weight,short-chain fatty acid concentration and reduced total bacterial count as well asγ-proteobacteria count and proportion(P<0.05).The OH-c treatment specifically increased full gastrointestinal tract weight and caecum contents weight compared to piglets fed CON and OH-f.Furthermore,OH-c reduced colonic crypt depth when compared to OH-f(P=0.018).In conclusion,supplementing OH to a diet for suckling piglets exerted subtle developmental effects on gastrointestinal morphology and colonic microbial community.These effects were largely independent from the particle size of the OH.

Intermittent suckling with or without comingling of non-littermate piglets before weaning improves piglet performance in the immediate post-weaning period when compared with conventional weaning

Background： In this experiment, intermittent suckling（IS） with or without the co-mingling（Co M） of piglets was studied as a method to stimulate solid feed intake and reduce post-weaning stress.Methods： Three weaning regimes using 30 multiparous sows were compared：（1） conventional weaning（CW）（n = 10 litters）, where piglets had continuous access to the sow until weaning（d 0, farrowing = d-25 relative to weaning）;（2） intermittent suckling（IS）（n = 10 litters）, where piglets were separated from the sow for 8 h/d starting at d-7（relative to weaning）; and（3） intermittent suckling with co-mingling（ISCo）（n = 10 litters） where IS started at d-7 and two litters were housed together during separation and then returned to their original sow. Ad libitum creep feed was available from d-17. At weaning pigs were housed in pens of 11 pigs, 27 pens in total. The ISCo treatment was divided in half to examine effects of different mixing strategies after weaning. Half of the ISCo litters were kept in familiar groups（ISCo F, familiar, n = 4） and the other half were mixed within treatment resulting in groups of unfamiliar pigs（ISCo NF, not familiar, n = 5）, the same as IS（n = 9） and CW（n = 9） treatments.Results： The ISCo piglets ate more creep feed in the week before weaning（P 〈 0.01）, but also showed more aggressive and manipulative behaviour on first day of Co M compared with CW piglets（P 〈 0.05）. IS with or without Co M increased exploratory and play behaviour on the first day of treatment intervention（P 〈 0.001） and increased sleeping behaviour on the last day of treatment intervention compared with CW（P 〈 0.001）. Mixing strategy at weaning had an effect on performance data with the highest growth and feed intake seen in ISCo F pigs 2 to 8 d after weaning（P 〈0.001）. IS and ISCo NF pigs also grew faster and ate more than CW pigs 2 to 8 d after weaning（P 〈 0.001）. Post-weaning injury scores suggested reduced aggression in ISCo as evidenced by reduced redness（skin irritation）（P 〈 0.05）, and a tendency for ISCo to have less scratches than CW（P 〈 0.1）. The IS pigs slept the most and displayed less manipulative behaviours on the day of weaning and plasma haptoglobin levels remained low in IS pigs after weaning（P ≤ 0.01）.Conclusions： Both intermittent suckling techniques improved production indices in the immediate post-weaning period. However, the addition of co-mingling before weaning in combination with grouping familiar pigs together after weaning improved performance in an additive manner.

Galacto-oligosaccharides improve barrier function and relieve colonic inflammation via modulating mucosa-associated microbiota composition in lipopolysaccharides-challenged piglets

Background:Galacto-oligosaccharides(GOS)have been shown to modulate the intestinal microbiota of suckling piglets to exert beneficial effects on intestinal function.However,the modulation of intestinal microbiota and intestinal function by GOS in intestinal inflammation injury models has rarely been reported.In this study,we investigated the effects of GOS on the colonic mucosal microbiota composition,barrier function and inflammatory response of lipopolysaccharides(LPS)-challenged suckling piglets.Methods:A total of 18 newborn suckling piglets were divided into three groups,the CON group,the LPS-CON group and the LPS-GOS group.Piglets in the LPS-GOS group were orally fed with 1 g/kg body weight of GOS solution every day.On the d 14,piglets in the LPS-CON and LPS-GOS group were challenged intraperitoneally with LPS solution.All piglets were slaughtered 2 h after intraperitoneal injection and sampled.Results:We found that the colonic mucosa of LPS-challenged piglets was significantly injured and shedding,while the colonic mucosa of the LPS-GOS group piglets maintained its structure.Moreover,GOS significantly reduced the concentration of malondialdehyde(MDA)and the activity of reactive oxygen species(ROS)in the LPS-challenged suckling piglets,and significantly increased the activity of total antioxidant capacity(T-AOC).GOS significantly increased the relative abundance of norank_f_Muribaculaceae and Romboutsia,and significantly decreased the relative abundance of Alloprevotella,Campylobacter and Helicobacter in the colonic mucosa of LPS-challenged suckling piglets.In addition,GOS increased the concentrations of acetate,butyrate and total short chain fatty acids(SCFAs)in the colonic digesta of LPS-challenged suckling piglets.GOS significantly reduced the concentrations of interleukin 1β(IL-1β),interleukin 6(IL-6),tumor necrosis factor-α(TNF-α)and cluster of differentiation 14(CD14),and the relative mRNA expression of Toll-like receptor 4(TLR4)and myeloid differentiation primary response 88(MyD88)in the LPS-challenged suckling piglets.In addition,GOS significantly reduced the relative mRNA expression of mucin2(MUC2),and significantly increased the protein expression of Claudin-1 and zonula occluden-1(ZO-1)in LPS-challenged suckling piglets.Conclusions:These results suggested that GOS can modulate the colonic mucosa-associated microbiota composition and improve the intestinal function of LPS-challenged suckling piglets.