Source of fiber influences growth,immune responses,gut barrier function and microbiota in weaned piglets fed antibiotic-free diets

This study examined the impacts of different fiber sources on growth, immune status and gut health in weaned piglets fed antibiotic-free diets. Sixty piglets (BW = 8.18 ± 1.35 kg) were assigned to 3 dietary treatments based on BW and gender in a randomized complete block design (5 replicates/treatment and 4 piglets [2 barrows and 2 gilts]/replicate): (1) an antibiotic-free diet (control, CON); (2) CON + 6% wheat bran (WB); (3) CON + 4% sugar beet pulp (SBP). Dietary WB supplementation tended to increase ADG compared with CON from d 1 to 14 (P = 0.051) and from d 1 to 28 (P = 0.099). Supplementation of WB increased (P < 0.05) G:F compared with CON and SBP from d 1 to 14 and from d 1 to 28. Compared with CON, the addition of WB reduced (P < 0.05) diarrhea rate from d 1 to 14 and tended (P = 0.054) to reduce diarrhea rate from d 1 to 28. The addition of WB decreased (P < 0.05) serum diamine oxidase activity on d 14, and up-regulated (P < 0.05) ileal mRNA levels of occludin on d 28 when compared with CON. Piglets fed WB showed decreased (P < 0.05) serum interleukin-6 levels compared to those fed SBP and decreased (P < 0.05) ileal interleukin-8 levels compared to those fed CON and SBP on d 28. Supplementation of WB increased (P < 0.05) serum levels of immunoglobulin A (IgA), IgG and IgM compared with SBP on d 14, and increased (P < 0.05) the levels of serum IgA and ileal sIgA compared with CON and SBP on d 28. Piglets fed WB showed an enhanced (P < 0.05) α-diversity of cecal microbiota than those fed SBP, while piglets fed SBP showed reduced (P < 0.05) α-diversity of cecal microbiota than those fed CON. Compared with CON, the addition of WB elevated (P < 0.05) the abundance of Lachnospira and cecal butyric acid level. Piglets fed WB also showed increased (P < 0.05) abundances of Lachnospira and unclassified_f_Lachnospiraceae compared with those fed SBP. Collectively, the supplementation of WB to antibiotic-free diets improved performance, immune responses, gut barrier function and microbiota compared with the CON and SBP fed piglets. Therefore, supplementing weaned piglets with WB was more effective than SBP.     

Serum biochemical parameters and amino acids metabolism are altered in piglets by early-weaning and proline and putrescine supplementations

The study was to investigate the effect of early-weaning stress and proline (Pro) and putrescine (Put) supplementations on serum biochemical parameters and amino acids (AA) metabolism in suckling and post-weaning pigs. Blood and small intestinal mucosa were harvested from suckling piglets at 1, 7, 14, and 21 d of age and piglets on d 1, 3, 5, and 7 after weaning at 14 d of age, as well as from piglets received oral administration of Pro and Put from 1 to 14 d old. In suckling piglets, the serum glucose, albumin and total cholesterol levels were increased (P < 0.05) with increasing age, whereas the serum globulin, urea nitrogen (BUN), alkaline phosphatase (ALP) and aspartate aminotransferase (AST) levels were lowered (P < 0.05). The concentrations of most serum AA and the AA transporters related gene expressions were highest in 7-d-old piglets (P < 0.05), whereas the phosphorylation status of the mammalian target of the rapamycin (mTOR) signaling pathway in the small intestine increased in piglets from 1 to 21 d old (P < 0.05). Weaning at 14 d old increased (P < 0.05) the BUN and triglycerides levels in serum, as well as jejunal solute carrier family 7 member 6 (SLC7A6), ileal SLC36A1 and SLC1A1 mRNA abundances at d 1 or 3 post-weaning. Weaning also inhibited (P < 0.05) the phosphorylation levels of mTOR and its downstream ribosomal protein S6 kinase 1 (S6K1) and 4E-binding protein-1 (4EBP1) in the small intestine of weanling pigs. Oral administration of Put and Pro decreased (P < 0.05) serum ALP levels and increased (P < 0.05) intestinal SLC36A1 and SLC1A1 mRNA abundances and mTOR pathway phosphorylation levels in post-weaning pigs. Pro but not Put treatment enhanced (P < 0.05) serum Pro, arginine (Arg) and glutamine (Gln) concentrations of weaning-pigs. These findings indicated that early-weaning dramatically altered the biochemical blood metabolites, AA profile and intestinal mTOR pathway activity, and Pro and Put supplementations improved the AA metabolism and transportation as well as activated the intestinal mTOR pathway in weanling-pigs. Our study has an important implication for the broad application of Pro and Put in the weaning transition of piglets.     

Different dietary protein sources influence growth performance,antioxidant capacity,immunity,fecal microbiota and metabolites in weaned piglets

The inclusion of high-quality proteins are commonly used in swine production.Our research investigated the effects of hydrolyzed wheat protein(HWP),fermented soybean meal(FSBM),and enzyme-treated soybean meal(ESBM)on growth performance,antioxidant capacity,immunity,fecal microbiota and metabolites of weaned piglets.A total of 144 piglets(weaned at 28 d)were allotted to 3 dietary treatments with 6 replicate pens per treatment and 8 piglets per pen.This study included 2 periods:d 0 to14 for phase 1 and d 15 to 28 for phase 2.Dietary treatments contained 15.90%HWP,15.80%FSBM,and 15.10%ESBM in phase 1,and 7.90%HWP,7.80%FSBM,and 7.50%ESBM in phase 2,respectively.The ADG of piglets in ESBM was increased(P<0.05)compared with HWP and FSBM during d 1e28.Compared with HWP and FSBM,ESBM increased(P<0.05)the ferric reducing ability of plasma(FRAP),and the serum level of superoxide dismutase(SOD)in piglets on d 14,as well as increased(P<0.05)the serum FRAP level in piglets on d 28.ESBM decreased(P<0.05)serum levels of DAO and IL-1b in piglets compared with HWP on d 28.ESBM enhanced(P<0.05)the relative abundance of Bacteroidetes,Oscillospiraceae and Christensenellaceae,as well as reduced the relative abundance of Clostridiaceae in the feces compared with HWP and FSBM.The PICRUSt analysis revealed that the number of gene tags related to degradation of valine,leucine and isoleucine,as well as lysine degradation in ESBM were lower(P<0.05)than that in HWP and FSBM.ESBM increased(P<0.05)the fecal butyrate level in piglets compared with FSBM,and ESBM tended to decrease(P=0.076)the fecal cadaverine level.Overall,ESBM had advantages over HWP and FSBM in improving antioxidant status,immune function,fecal bacteria and metabolites for weaned piglets.     

Early life administration of milk fat globule membrane promoted SCFA-producing bacteria colonization,intestinal barriers and growth performance of neonatal piglets

Milk fat globule membrane (MFGM) possesses various nutritional and biological benefits for mammals, whereas its effects on neonatal gut microbiota and barrier integrity remained unclear. This study investigated the effects of MFGM administration on microbial compositions and intestinal barrier functions of neonatal piglets. Sixteen newborn piglets were randomly allocated into a CON group or MFGM group, orally administered with saline or MFGM solution (1 g/kg body weight) respectively during the first postnatal week, and all piglets were breastfed during the whole neonatal period. The present study found that the MFGM oral administration during the first postnatal week increased the plasma immunoglobulin (Ig) G level, body weight and average daily gain of piglets (P < 0.05) on 21 d. Additionally, MFGM administration enriched fecal SCFA-producing bacteria (Ruminococaceae_UCG-002, Ruminococaceae_UCG-010, Ruminococaceae_UCG-004, Ruminococaceae_UCG-014 and [Ruminococcus]_gauvrearuii_group), SCFA concentrations (acetate, propionate and butyrate; P < 0.05) and their receptor (G-protein coupled receptor 41, GPR41). Furthermore, MFGM administration promoted intestinal villus morphology (P < 0.05) and barrier functions by upregulating genes of tight junctions (E-cadherin, claudin-1, occludin and zonula occludin 1 [ZO-1]), mucins (mucin-13 and mucin-20) and interleukin (IL)-22 (P < 0.05). Positive correlation was found between the beneficial microbes and SCFA levels pairwise with the intestinal barrier genes (P < 0.05). In conclusion, orally administrating MFGM during the first postnatal week stimulated SCFA-producing bacteria colonization and SCFA generation, enhanced intestinal barrier functions and consequently improved growth performance of neonatal piglets on 21 d. Our findings will provide new insights about MFGM intervention for microbial colonization and intestinal development of neonates during their early life.     

Gut microbiota absence and transplantation affect growth and intestinal functions: An investigation in a germ-free pig model

This study was conducted to investigate host–microbiota interactions and explore the effects of maternal gut microbiota transplantation on the growth and intestinal functions of newborns in a germ-free (GF) pig model. Twelve hysterectomy-derived GF Bama piglets were reared in 6 sterile isolators. Among them, 6 were considered as the GF group, and the other 6 were orally inoculated with healthy sow fecal suspension as fecal microbiota transplanted (FMT) group. Another 6 piglets from natural birth were regarded as the conventional (CV) group. The GF and FMT groups were hand-fed with Co60-γ-irradiated sterile milk powder, while the CV group was reared by lactating Bama sows. All groups were fed for 21 days. Then, all piglets and then were switched to sterile feed for another 21 days. Results showed that the growth performance, nutrient digestibility, and concentrations of short-chain fatty acids in the GF group decreased (P < 0.05). Meanwhile, the serum urea nitrogen concentration and digesta pH values in the GF group increased compared with those in the FMT and CV groups (P < 0.05). Compared with the CV group, the GF group demonstrated upregulation in the mRNA expression levels of intestinal barrier function-related genes in the small intestine (P < 0.05). In addition, the mRNA abundances of intestinal development and absorption-related genes in the small intestine and colon were higher in the GF group than in the CV and FMT groups (P < 0.05). The FMT group exhibited greater growth performance, lipase activity, and nutrient digestibility (P < 0.05), higher mRNA expression levels of intestinal development and barrier-related genes in the small intestine (P < 0.05), and lower mRNA abundances of pro-inflammatory factor in the colon and jejunum (P < 0.05) than the CV group. In conclusion, the absence of gut microbes impaired the growth and nutrient digestibility, and healthy sow gut microbiota transplantation increased the growth and nutrient digestibility and improved the intestinal development and barrier function of newborn piglets, indicating the importance of intestinal microbes for intestinal development and functions.     

Effect of dietary folate level on organ weight,digesta pH,short-chain fatty acid concentration,and intestinal microbiota of weaned piglets

Folate is increasingly thought to promote gastrointestinal health and regulate the diversity of gut microbiota to alleviate weaning stress in piglets. The present study was conducted to investigate the effects of folate on organ weight, digesta pH, short-chain fatty acids (SCFAs) concentration, and intestinal microbiota in weaned piglets. A total of 28 piglets (6.73 ± 0.62 kg) were allocated to four dietary treatments consisting of a control group, 3, 9, and 18 mg/kg of folate supplementation in a 14-d feeding trial. The results showed that piglets fed with 9 and 18 mg/kg of folate supplementation had greater (P < 0.05) average liver and spleen weight than the control group. Folate supplementation (9 and 18 mg/kg) can significantly increase (P < 0.05) the stomach pH and tend (P < 0.10) to decrease the cecum pH. Folate treatment (9 and 18 mg/kg) had a positive effect on the metabolism of SCFAs in piglets, in particular, compared with the control group, and the content of acetic acid (AA) and valeric acid was markedly increased (P < 0.05) in the cecum and colon, respectively. Moreover, isobutyric acid, butyric acid, and isovaleric acid were tended (P < 0.10) to increase in the colon. Cecum contents samples were used to determine bacterial community diversity by 16S rRNA gene amplicon sequencing. At the genus level, in the cecum, there was a higher (P < 0.05) relative abundance of Lactobacillus reuteri, Lactobacillus salivarius, and Lactobacillus mucosae in the 9 mg/kg folate supplementation group. The functional pathways analysis predicted that folate may modify nutrient metabolism by changing the gut microbiota function of weaned piglets. Furthermore, the data showed that Lactobacillus was positively correlated with AA in the cecum. Overall, these findings suggested that folate treatment could increase the organ weight and the stomach pH of weaned piglets and had beneficial effects on gut health, which might be attributed to the alteration in intestinal microbiota induced by folate and the interaction of the intestinal microbiota with SCFAs.     

Changes in progenitors and differentiated epithelial cells of neonatal piglets

This study aimed to assess the changes of small intestinal morphology,progenitors,differentiated epithelial cells,and potential mechanisms in neonatal piglets.Hematoxylin and eosin staining of samples from 36 piglets suggested that dramatic changes were observed in the jejunum crypts depth and crypt fission index of neonatal piglets(P<0.001).The number of intestinal stem cells(ISC)tended to increase(P<0.10),and a decreased number of enteroendocrine cells appeared in the jejunal crypt on d 7(P<0.05).Furthermore,the mRNA expression of jejunal chromogranin A(ChgA)was down-regulated in d 7 piglets(P<0.05).There was an up-regulation of the adult ISC marker gene of SPARC related modular calcium binding 2(Smoc2),and Wnt/b-catenin target genes on d 7(P<0.05).These results were further verified in vitro enteroid culture experiments.A mass of hollow spheroids was cultured from the fetal intestine of 0-d-old piglets(P<0.001),whereas substantial organoids with budding and branching structures were cultured from the intestine of 7-d-old piglets(P<0.001).The difference was reflected by the organoid budding efficiency,crypt domains per organoid,and the surface area of the organoid.Furthermore,spheroids on d 0 had more Ki67-positive cells and enteroendocrine cells(P<0.05)and showed a decreasing trend in the ISC and goblet cells(P<0.10).Moreover,the mRNA expression of spheroids differed markedly from that of organoids,with low expression of intestinal differentiation gene(Lysozyme;P<0.05),epithelial-specific markers(Villin,E-cadherin;P<0.05),and adult ISC markers(leucine-rich repeat-containing G protein-coupled receptor 5[Lgr5],Smoc2;P<0.001),and upregulation of fetal marker(connexin 43[Cnx43];P<0.05).The mRNA expression of relevant genes was up-regulated,and involved in Wnt/b-catenin,epidermal growth factor(EGF),Notch,and bone morphogenetic protein(BMP)signaling on d 7 organoids(P<0.05).Spheroids displayed low differentiated phenotype and high proliferation,while organoids exhibited strong differentiation potential.These results indicated that the conversion from the fetal progenitors(spheroids)to adult ISC(normal organoids)might largely be responsible for the fast development of intestinal epithelial cells in neonatal piglets.     

Organic acid blends improve intestinal integrity,modulate short-chain fatty acids profiles and alter microbiota of broilers under necrotic enteritis challenge

Controlling enteric diseases of broilers is crucial.Among many additives,organic acids(OA)and their blends are gaining attention to combat diseases in the post-antibiotic era.The current study evaluated the potentials of short-chain fatty acids(SCFA)and medium-chain fatty acids(MCFA)blends and/or phenolic compounds on intestinal integrity,intestinal pH,caecal microbiota,and caecal SCFA profiles of broilers under necrotic enteritis(NE)challenge.The additives used were:(A)a blend of SCFA,MCFA,and a phenolic compound(SMP),(B)a blend of free and buffered SCFA with MCFA(SMF),and(C)a blend of free and buffered SCFA with a high concentration of MCFA(SHM).A total of 1,404 male parental chicks of Ross 308 broilers were randomly allocated to 78 floor pens on hatching day with 6 treatments replicated 13 times with 18 birds per pen.The treatments were:UCC,unchallenged control;CHC,challenged control;BAC,challenged group plus zinc bacitracin;SMP,challenged group plus additive SMP;SMF,challenged group plus additive SMF;SHM,challenged group plus additive SHM.Birds were challenged with field-strain Eimeria spp.on d 9 and Clostridium perfringens on d 14.Birds challenged with NE increased fluorescein isothiocyanate dextran(FITC-d)concentration in serum,reduced acetate and butyrate concentrations,and increased Bacteroides and C.perfringens load in the caeca(P<0.05).Birds fed additives decreased FITC-d from gut to serum,reduced Bacteroides(d 16,P<0.05)and numerically reduced C.perfringens load compared to CHC group.Birds fed additive SHM had higher concentrations of acetate and butyrate(d 21,P<0.05)than CHC group but were not different from SMP and SMF groups.All the additives exhibited similar intestinal protection against NE compared to the BAC group indicated by FITC-d concentration in serum,acetate,propionate and butyrate concentrations in the caeca,and caecal bacterial loads except for the C.perfringens(P>0.05).The SMP group had a higher load compared to BAC(P<0.05).These findings suggest the promising effects of OA blends as alternatives to BAC to ameliorate the impact of NE challenge of broilers as indicated by improved intestinal health.     

Interaction between xylanase and a proton pump inhibitor on broiler chicken performance and gut function

Three hundred thirty-six Ross 308 male broiler chicks were used in a 21-d study to explore performance and gut function when treated with a proton pump inhibitor(PPI;0 or 89 mg/kg)in a 2×2 factorial arrangement with a xylanase(Xyl;0 or 0.1 g/kg)to determine if the beneficial activity of arabinoxylan(AX)depolymerisation,through arabinoxylo-oligosaccharides(AXOS)production,starts in the upper gastrointestinal tract.Treatment with the PPI started from d 14,and by d 21 animal performance had deteriorated(P<0.001).An interaction was observed between PPI and Xyl for feed conversion ratio(FCR)(P<0.05),whereby the combination reduced the negative effect of PPI on FCR.Application of PPI raised digesta pH in the gizzard and caecum(P<0.05),increased protein concentrations in the lower gut(P<0.05)and reduced intake of digestible nutrients(P<0.05).Caecal concentrations of indole,p-cresol,ammonia and the ratio of total volatile fatty acid(VFA)to butyric acid were increased with PPI(P<0.05),indicating enhanced protein fermentation.Xylanase activity in the digesta were greatest in the caeca,especially when Xyl was supplemented(P<0.001).The concentration of total soluble AX was greater in the gizzard and ileal digesta with Xyl supplementation(P<0.05),supporting the depolymerisation action of xylanase even under acidic conditions.These data suggest xylanase may function in the gizzard even though pH is not optimal for activity and emphasises the importance of chlorohydric acid secretions in ensuring overall optimum gut function.AX depolymerisation benefits animal performance although it is still unknown how the AXOS produced with xylanase supplementation in the upper gastrointestinal tract influence the microbial populations and overall gut functionality.     

N-Acetyl-D-glucosamine improves the intestinal development and nutrient absorption of weaned piglets via regulating the activity of intestinal stem cells

Early weaning in piglets can cause a series of negative effects.This causes serious losses to the livestock industry.N-Acetyl-D-glucosamine(D-GlcNAc)plays an important role in regulating the homeostasis of the intestine.This study aimed to investigate the effects of D-GlcNAc on the growth performance and intestinal function of weaned piglets.Twenty-four weaned piglets([Yorkshire×Landrace]
Duroc,6.58±0.15 kg,n=8)at 21 d old were fed 3 diets supplemented with 0(control),1 and 3 g/kg D-GlcNAc.The intestinal organoid model was used to verify the regulatory mechanism of D-GlcNAc on intestinal epithelial cells.On the whole,supplementation of D-GlcNAc in the piglet diet has no significant effect on the growth performance and diarrhoea of weaned piglets(P>0.05).The apparent digestibility of nutrients and mRNA abundance of nutrient transporters in the 1 g/kg D-GlcNAc group were increased significantly(P<0.05).D-GlcNAc did not affect villus height(VH)and crypt depth(CD)but resulted in a numerically shorter VH and shallower CD,which lead to an increase in ileal VH:CD ratio(P<0.05).Cell shedding rates in the ileum villi increased(P<0.05).The relative length and weight of the small intestine of weaned piglets increased(P<0.05).In vitro studies found that the budding rates of organoids treated with 0.1 mmol/L D-GlcNAc increased on the d 3 and 5(P<0.05).The average budding numbers per budding organoid treated with 0.1 and 10 mmol/L D-GlcNAc increased on d 3(P<0.05).D-GlcNAc upregulated leucine rich repeat containing G protein-coupled receptor 5(Lgr5⁺)and Chromogranin A mRNA abundance in organoids(P<0.05).Mucin 2(Muc2)expression increased when treated with 1 and 10 mmol/L D-GlcNAc(P<0.05).In conclusion,dietary D-GlcNAc cannot improve the growth performance of weaned piglets.However,it can promote the growth and development of the intestinal tract and improve the digestion and absorption capacity of the intestine,which is achieved by affecting the activity of intestinal stem cells.     

Natural capsicum extract replacing chlortetracycline enhances performance via improving digestive enzyme activities,antioxidant capacity,anti-inflammatory function,and gut health in weaned pigs

The objective of this study was to investigate the effects of natural capsicum extract (NCE, containing 2% natural capsaicin, the rest is carrier) replacing chlortetracycline (CTC) on performance, digestive enzyme activities, antioxidant capacity, inflammatory cytokines, and gut health in weaned pigs. A total of 108 weaned pigs (Duroc × [Landrace × Yorkshire], initial body weight = 8.68 ± 1.34 kg; weaned on d 28) were randomly allotted into 3 treatments with 6 replicate pens per treatment (3 barrows and 3 gilts per pen). The treatments include a corn-soybean meal basal diet as a control group (CON), a CTC group (basal diet + CTC at 75 mg/kg), and a NCE group (basal diet + NEC at 80 mg/kg). Compared with CON and CTC, NCE had increased (P < 0.05) average daily gain in phase 2 (d 15 to 28) and overall (d 1 to 28), and higher (P < 0.05) apparent total tract digestibility of gross energy, dry matter, crude protein, and organic matter in phase 1 (d 1 to 14). These pigs also had increased (P < 0.05) pancrelipase activity in pancreas, α-amylase, lipase and protease activities in the jejunal mucosa, and lipase activity in the ileal mucosa on d 28. Moreover, NCE had increased (P < 0.05) the contents of growth hormone, β-endorphin, 5-hydroxytryptamine, total antioxidant capacity, total superoxide dismutase, catalase, and IL-10, as well as decreased (P < 0.05) contents of malondialdehyde, tumor nuclear factor-α, interferon-γ, and interleukin-6 in serum on d 28 compared with CON and CTC. NCE showed higher (P < 0.05) propionic acid, butyric acid and total volatile fatty acids (VFA) contents, and increased (P < 0.05) relative abundance of Faecalibacterium in colon, as well as higher (P < 0.05) propionic acid and total volatile fatty acids in cecum on d 28 compared with CON. In conclusion, NCE replacing CTC could enhance performance via improving digestive enzyme activities, antioxidant capacity, anti-inflammatory function, gut VFA composition and microbiota community in weaned pigs, and it could be used as a potential target for the development of feed additives.     

Dietary nano-selenium alleviated intestinal damage of juvenile grass carp(Ctenopharyngodon idella)induced by high-fat diet:Insight from intestinal morphology,tight junction,inflammation,anti-oxidization and intestinal microbiota

In recent years,high-fat diet(HFD)has been widely applied in aquaculture,which reduces the intestinal health of cultured fish.The current study evaluated the protective effects of nano-selenium(nano-Se)on intestinal health of juvenile grass carp(Ctenopharyngodon idella)fed with HFD.A total of 135 experimental fish were fed with a regular diet(Con),a HFD(HFD)and a HFD containing nano-Se at 0.6 mg/kg(HSe)for 10 weeks.The results showed that dietary nano-Se significantly improved the survival rate and feed efficiency which were reduced by HFD in juvenile grass carp(P<0.05).Also,nano-Se(0.6 mg/kg)supplement alleviated intestinal damage caused by the HFD,thus maintaining the integrity of the intestine.Moreover,it significantly up-regulated the expression of genes related to tight junction(ZO-1,claudin-3 and occludin),anti-oxidization(GPx4a and GPx4b),and the protein of ZO-1 in the intestine of juvenile grass carp,which were depressed by the HFD(P<0.05).Furthermore,nano-Se supplementation significantly suppressed the expressions of genes related to the inflammation,including inflammatory cytokines(IL-8,IL-1β,IFN-γ,TNF-αand IL-6),signaling molecules(TLR4,p38 MAPK and NF-kB p65),and protein expression of NF-kB p65 and TNF-αin the intestine of juvenile grass carp which were induced by the HFD(P<0.05).Besides,dietary nano-Se normalized the intestinal microbiota imbalance of juvenile grass carp caused by the HFD through increasing the abundance of the beneficial bacteria,e.g.,Fusobacteria.Finally,dietary nano-Se increased the production of short chain fatty acids(SCFA)in the intestine,especially for butyric acid and caproic acid,which were negatively related to the increase of intestinal permeability and inflammation.In summary,supply of nano-Se(0.6 mg/kg)in HFD could effectively alleviate intestinal injury of juvenile grass carp by improving intestinal barrier function and reducing intestinal inflammation and oxidative stress.These positive effects may be due to the regulation of nano-Se on intestinal microbiota and the subsequently increased beneficial SCFA levels.     

Growth performance,fatty acid composition,and lipid metabolism are altered in groupers(Epinephelus coioides)by dietary fish oil replacement with palm oil

In this study,we conducted a 56-d feeding trial to investigate the effects of replacing the fish oil(FO)with palm oil(PO)on the performance,tissue fatty acid(FA)composition,and mRNA levels of genes related to hepatic lipid metabolism in grouper(Epinephelus coioides).Five isolipidic(13%crude lipid)and isonitrogenous(48%CP)diets were formulated by incrementally adding PO to the control diet(25%fish meal and 9%added FO)to replace FO in the control diets.Triplicate groups of 30 groupers(initial weight:12.6±0.1 g)were fed one of the diets twice daily,to apparent satiety.The replacement of FO with 50%PO revealed maximum growth without affecting the performance and whole-body proximate compositions,and replacing FO with 100%PO revealed a comparable(P>0.05)growth with that of the control diet,suggesting PO as a suitable alternative to FO.The analysis of FA profiles in the dorsal muscle and liver though reflected the FA profile of the diet,PO substitutions above 50%could compromise(P<0.05)the FA profile in the liver and flesh of the fish species in comparison with the control diet.Furthermore,the mRNA levels of FAS,G6PD,LPL,PPARА,and D6FAD genes in the liver had positive linear and/or quadratic responses,but the SCD,HSL,ATGL,FABP,SREBP-1C and ELOVL5 had the opposite trend,with increasing dietary PO inclusion levels,whereas the mRNA level of ACC was not affected by dietary treatments.The optimal level of PO substitution for FO was estimated to be 47.1%of the feed,based on the regression analysis of percent weight gains against dietary PO inclusion levels;however,it might affect the FA profile in the liver and flesh of the fish species,and further study is required to investigate whether the changes in tissue FA composition will affect the welfare and market value over a production cycle of grouper.     

Prevotella-rich enterotype may benefit gut health in finishing pigs fed diet with a high amylose-to-amylopectin ratio

To investigate the influence of baseline enterotypes and dietary starch type on the concentration of short-chain fatty acids (SCFA), numbers of butyrate producing bacteria and the expression of genes related to intestinal barrier and inflammatory response in the colon of finishing pigs, a 60-d in vivo trial was conducted. A 2-wk pre-trial with 102 crossbred (Duroc × [Landrace × Yorkshire]) finishing barrows (90 d old) was conducted to screen enterotypes. Then, a total of 32 pigs (87.40 ± 2.76 kg) with high (HPBR, ≥ 14) and low (LPBR, ≤ 2) Prevotella-to-Bacteroides ratios (PBR) in equal measure were selected and randomly divided into 4 groups with 8 replicates per group and 1 pig per replicate. The trial was designed following a 2 (PBR) × 2 (amylose-to-amylopectin ratio, AMR) factorial arrangement. Pigs with different PBR were fed diets based on corn-soybean meal with high AMR (HAMR, 1.24) or low AMR (LAMR, 0.23), respectively. Results showed that neither PBR nor AMR influenced the growth performance of pigs. HPBR pigs fed HAMR diet had a higher number of colonic Clostridium cluster XIVa and higher gene expression of butyrate kinase compared to the LPBR pigs (P < 0.05). The HPBR pigs fed HAMR diets also had increased colonic concentrations of total SCFA and propionate compared to the LPBR pigs (P < 0.05). Comparing with other pigs, HPBR pigs fed HAMR diets showed a lower (P < 0.05) expression of histone deacetylases (HDAC) gene and higher (P < 0.05) expression of G protein-coupled receptor 43 gene (GPR 43) in the colonic mucosa. The interaction (P < 0.05) of HPBR and HAMR was also found to decrease the gene expression of interleukin (IL)-6, IL-12, IL-1β and tumor necrosis factor-α (TNF-α) in colonic mucosa. These findings show that HAMR diet increased the abundance and activity of butyrate-producing bacteria and the concentration and absorption of SCFA, which may be associated with the decreased gene expression of inflammatory cytokines in the colonic mucosa of pigs with Prevotella-rich enterotype. All these alterations are likely to have a positive effect on the intestinal health of finishing pigs.     

Dietary chenodeoxycholic acid improves growth performance and intestinal health by altering serum metabolic profiles and gut bacteria in weaned piglets

Nutritional diarrhea and subsequent performance degradation in weaned piglets are major challenges for the pig industry. Bile acids (BA) can be added to the diet as emulsifiers. This experiment was conducted to investigate the effects of chenodeoxycholic acid (CDCA), a major primary BA, on growth performance, serum metabolic profiles and gut health in weaned piglets. A total of 72 healthy weaned piglets were randomly assigned to the control (CON) and the CDCA groups, which were feed a basal diet and the basal diet supplemented with 200 mg/kg CDCA for 30 d, respectively. Our results demonstrated that CDCA significantly increased final BW and average daily gain (ADG), decreased feed-to-gain (F:G) ratio and tended to reduce diarrhea incidence. In addition, CDCA increased the villus height-to-crypt depth (V:C) ratio, elevated goblet cell numbers and the expression of tight junction proteins, suggesting the enhancement of intestinal barrier function. As an emulsifier, CDCA increased jejunal lipase activity and the mRNA expression of pancreatic lipases. CDCA supplementation also altered the serum metabolic profiles, including increasing the levels of indole 3-acetic acid, N′-formylkynurenine and theobromine that were beneficial for gut health. Moreover, the relative abundance of 2 beneficial gut bacteria, Prevotella9 and PrevotellaceaeTCG-001, were increased, whereas the relative abundance of a harmful bacteria, Dorea, was decreased in the gut of weaned piglets supplemented with CDCA. Importantly, the altered serum metabolic profiles showed a strong correlation with the changed gut bacteria. In conclusion, CDCA improved the growth performance of weaned piglets by improving intestinal morphology and barrier function, and enhancing lipid digestion, accompanied by alterations of serum metabolic profiles, and changes in relative abundance of certain gut bacteria.     

Responsive changes of rumen microbiome and metabolome in dairy cows with different susceptibility to subacute ruminal acidosis

Subacute ruminal acidosis(SARA)represents one of the most important digestive disorders in intensive dairy farms,and dairy cows are individually different in the severity of SARA risk.The objectives of the current study were to investigate differences in the ruminal bacterial community and metabolome in dairy cattle with different susceptibility to SARA.In the present study,12 cows were initially enrolled in the experiment.Based on average ruminal pH,4 cows with the lowest ruminal pH were assigned to the susceptible group(SUS,pH=5.76,n=4)and 4 cows with the highest ruminal pH assigned to the tolerant group(TOL,pH=6.10,n=4).Rumen contents from susceptible(SUS,n=4)and tolerant(TOL,n=4)dairy cows were collected through rumen fistula to systematically reveal the rumen microbial and metabolic alterations of dairy cows with different susceptibility to SARA using multi-omics approaches(16S and 18S rRNA gene sequencing and metabolome).The results showed that despite being fed the same diet,SUS cows had lower ruminal pH and higher concentrations of total volatile fatty acids(VFA)and propionate than TOL cows(P<0.05).No significant differences were observed in dry matter intake,milk yield,and other milk compositions between the SUS and TOL groups(P>0.05).The principal coordinates analysis based on the analysis of molecular variance indicated a significant difference in bacterial composition between the two groups(P=0.01).More specifically,the relative abundance of starch-degrading bacteria(Prevotella spp.)was greater(P<0.05),while the proportion of fiber-degrading bacteria(unclassified Ruminococcaceae spp.,Ruminococcus spp.,Papillibacter,and unclassified Family_-XIII)was lower in the rumen of SUS cows compared with TOL cows(P<0.05).Community analysis of protozoa showed that there were no significant differences in the diversity,richness,and community structure(P>0.05).Metabolomics analysis revealed that the concentrations of organic acids(such as lactic acid),biogenic amines(such as histamine),and bacterial degradation products(such as hypoxanthine)were significantly higher in the SUS group compared to the TOL group(P<0.05).These findings revealed that the higher proportion of starch-degrading bacteria/lower fiber-degrading bacteria in the rumen of SUS cows resulted in higher VFA-producing capacity,in particular propionate.This caused a disruption in metabolic homeostasis in the rumen which might be the reason for the higher susceptibility to SARA.Overall,these findings enhanced our understanding of the ruminal microbiome and metabolic changes in cows susceptible to SARA.     

Dietary inclusion of multispecies probiotics to reduce the severity of post-weaning diarrhea caused by Escherichia coli F18+ in pigs

This study was aimed to determine the efficacy of multispecies probiotics in reducing the severity of post-weaning diarrhea caused by enterotoxigenic Escherichia coli (ETEC) F18⁺ on newly weaned pigs. Thirty-two pigs (16 barrows and 16 gilts, BW = 6.99 ± 0.33 kg) at 21 d of age were individually allotted in a randomized complete block design with 2 × 2 factorial arrangement of treatments. Pigs were selected from sows not infected previously and not vaccinated against ETEC. Pigs were fed experimental diets for 25 d based on 10 d phase 1 and 15 d phase 2. The factors were ETEC challenge (oral inoculation of saline solution or E. coli F18⁺ at 2 × 10⁹ CFU) and probiotics (none or multispecies probiotics 0.15% and 0.10% for phase 1 and 2, respectively). Body weight and feed intake were measured on d 5, 9, 13, 19, and 25. Fecal scores were measured daily. Blood samples were taken on d 19 and 24. On d 25, all pigs were euthanized to obtain samples of digesta, intestinal tissues, and spleen. The tumor necrosis factor alpha (TNFα), malondialdehyde (MDA), peptide YY (PYY), and neuropeptide Y (NPY) were measured in serum and intestinal tissue. Data were analyzed using the MIXED procedure of SAS. The fecal score of pigs was increased (P < 0.05) by ETEC challenge at the post–challenge period. The ETEC challenge decreased (P < 0.05) jejunal villus height and crypt depth, tended to increase (P = 0.056) jejunal TNFα, increased (P < 0.05) ileal crypt depth, and decreased (P < 0.05) serum NPY. The probiotics decreased (P < 0.05) serum TNFα, tended to reduce (P = 0.064) jejunal MDA, tended to increase (P = 0.092) serum PYY, and increased (P < 0.05) jejunal villus height, and especially villus height-to-crypt depth ratio in challenged pigs. Growth performance of pigs were not affected by ETEC challenge, whereas the probiotics increased (P < 0.05) ADG and ADFI and tended to increase (P = 0.069) G:F ratio. In conclusion, ETEC F18⁺ challenge caused diarrhea, intestinal inflammation and morphological damages without affecting the growth performance. The multispecies probiotics enhanced growth performance by reducing intestinal inflammation, oxidative stress, morphological damages.     

Dietary dihydroartemisinin supplementation alleviates intestinal inflammatory injury through TLR4/NOD/NF-κB signaling pathway in weaned piglets with intrauterine growth retardation

The aim of present study was to evaluate whether diets supplemented with dihydroartemisinin (DHA) could alleviate intestinal inflammatory injury in weaned piglets with intrauterine growth retardation (IUGR). Twelve normal birth weight (NBW) piglets and 12 piglets with IUGR were fed a basal diet (NBW-CON and IUCR-CON groups), and another 12 piglets with IUGR were fed the basal diet supplemented with DHA at 80 mg/kg (IUGR-DHA group) from 21 to 49 d of age. At 49 d of age, 8 piglets with similar body weight in each group were sacrificed. The jejunal and ileal samples were collected for further analysis. The results showed that IUGR impaired intestinal morphology, increased intestinal inflammatory response, raised enterocyte apoptosis and reduced enterocyte proliferation and activated transmembrane toll-like receptor 4 (TLR4)/nucleotide-binding and oligomerization domain (NOD)/nuclear factor-κB (NF-κB) signaling pathway. Dihydroartemisinin inclusion ameliorated intestinal morphology, indicated by increased villus height, villus height-to-crypt depth ratio, villus surface area and decreased villus width of piglets with IUGR (P < 0.05). Compared with NBW piglets, IUGR piglets supplemented with DHA exhibited higher apoptosis index and caspase-3 expression, and lower proliferation index and proliferating cell nuclear antigen expression in the intestine (P < 0.05). Dihydroartemisinin supplementation attenuated the intestinal inflammation of piglets with IUGR, indicated by increased concentrations of intestinal inflammatory cytokines and lipopolysaccharides (P < 0.05). In addition, DHA supplementation down-regulated the related mRNA expressions of TLR4/NOD/NF-κB signaling pathway and upregulated mRNA expressions of negative regulators of TLR4 and NOD signaling pathway in the intestine of piglets with IUGR (P < 0.05). Piglets in the IUGR-DHA group showed lower protein expressions of TLR4, phosphorylated NF-κB (pNF-κB) inhibitor α, nuclear pNF-κB, and higher protein expression of cytoplasmic pNF-κB in the intestine than those in the IUGR-CON group (P < 0.05). In conclusion, DHA supplementation could improve intestinal morphology, regulate enterocyte proliferation and apoptosis, and alleviate intestinal inflammation through TLR4/NOD/NF-κB signaling pathway in weaned piglets with IUGR.