The protective role of prebiotics and probiotics on diarrhea and gut damage in the rotavirus-infected piglets

Rotavirus is one of the pathogenic causes that induce diarrhea in young animals, especially piglets, worldwide. However, nowadays, there is no specific drug available to treat the disease, and the related vaccines have no obvious efficiency in some countries. Via analyzing the pathogenesis of rotavirus, it inducing diarrhea is mainly due to disturb enteric nervous system, destroy gut mucosal integrity, induce intracellular electrolyte imbalance, and impair gut microbiota and immunity. Many studies have already proved that prebiotics and probiotics can mitigate the damage and diarrhea induced by rotavirus infection in hosts. Based on these, the current review summarizes and discusses the effects and mechanisms of prebiotics and probiotics on rotavirus-induced diarrhea in piglets. This information will highlight the basis for the swine production utilization of prebiotics and probiotics in the prevention or treatment of rotavirus infection in the future.

All-trans retinoic acid alleviates transmissible gastroenteritis virus-induced intestinal inflammation and barrier dysfunction in weaned piglets

Background Transmissible gastroenteritis virus(TGEV)is one of the main pathogens causing severe diarrhea of pig-lets.The pathogenesis of TGEV is closely related to intestinal inflammation.All-trans retinoic acid(ATRA)is the main active metabolite of vitamin A,which has immunomodulatory and anti-inflammatory properties.However,it is unclear whether ATRA can alleviate TGEV-induced intestinal inflammation and barrier dysfunction in piglets.This study aimed to investigate the effects of ATRA on growth performance,diarrhea,intestinal inflammation and intesti-nal barrier integrity of TGEV-challenged piglets.Methods In a 19-d study,32 weaned piglets were randomly divided into 4 treatments:Control group(basal diet),TGEV group(basal diet+TGEV challenge),TGEV+ATRA5 group(basal diet+5 mg/d ATRA+TGEV challenge)and TGEV+ATRA15 group(basal diet+15 mg/d ATRA+TGEV challenge).On d 14,piglets were orally administered TGEV or the sterile medium.Results Feeding piglets with 5 and 15 mg/d ATRA alleviated the growth inhibition and diarrhea induced by TGEV(P<0.05).Feeding piglets with 5 and 15 mg/d ATRA also inhibited the increase of serum diamine oxidase(DAO)activ-ity and the decrease of occludin and claudin-1 protein levels in jejunal mucosa induced by TGEV,and maintained intestinal barrier integrity(P<0.05).Meanwhile,5 mg/d ATRA feeding increased the sucrase activity and the expres-sions of nutrient transporter related genes(GLUT2 and SLC7A1)in jejunal mucosa of TGEV-challenged piglets(P<0.05).Furthermore,5 mg/d ATRA feeding attenuated TGEV-induced intestinal inflammatory response by inhibit-ing the release of interleukin(IL)-1β,IL-8 and tumor necrosis factor-α(TNF-α),and promoting the secretion of IL-10 and secretory immunoglobulin A(sIgA)(P<0.05).Feeding 5 mg/d ATRA also down-regulated the expressions of Toll-like receptors and RIG-I like receptors signaling pathway related genes(TLR3,TLR4,RIG-I,MyD88,TRIF and MAVS)and the phosphorylation level of nuclear factor-κB-p65(NF-κB p65),and up-regulated the inhibitor kappa B alpha(IκBα)protein level in jejunal mucosa of TGEV-challenged piglets(P<0.05).Conclusions ATRA alleviated TGEV-induced intestinal barrier damage by inhibiting inflammatory response,thus improving the growth performance and inhibiting diarrhea of piglets.The mechanism was associated with the inhibi-tion of NF-κB signaling pathway mediated by TLR3,TLR4 and RIG-I.

Yeast hydrolysate attenuates lipopolysaccharide-induced inflammatory responses and intestinal barrier damage in weaned piglets

Background Intestinal inflammation is the main risk factor causing intestinal barrier dysfunction and lipopolysaccharide(LPS)can trigger inflammatory responses in various eukaryotic species.Yeast hydrolysate(YH)possesses multibiological effects and is received remarkable attention as a functional ingredient for improving growth performance and promoting health in animals.However,there is still inconclusive on the protective effects of dietary YH supplementation on intestinal barrier of piglets.This study was conducted to investigate the attenuate effects of YH supplementation on inflammatory responses and intestinal barrier injury in piglets challenged with LPS.Methods Twenty-four piglets(with an average body weight of 7.42±0.34 kg)weaned at 21 days of age were randomly assigned to one of two dietary treatments(12 replications with one pig per pen):a basal diet or a basal diet containing YH(5 g/kg).On the 22nd d,6 piglets in each treatment were intraperitoneally injected with LPS at 150μg/kg BW,and the others were injected with the same amount of sterile normal saline.Four hours later,blood samples of each piglet were collected and then piglets were euthanized.Results Dietary YH supplementation increased average daily feed intake and average daily gain(P<0.01),decreased the ratio of feed intake to gain of piglets(P sponse,evidenced by the increase o=0.048).Lipopolysaccharide(LPS)injection induced systemic inflammatory ref serum concentrations of haptoglobin(HP),adrenocorticotropic hormone(ACTH),cortisol,and interleukin-1β(IL-1β).Furthermore,LPS challenge resulted in inflammatory intestinal damage,by up-regulation of the protein or mRNA abundances of tumor necrosis factor-α(TNF-α),IL-1β,toll-like receptors 4(TLR4)and phosphor-nuclear factor-κB-p65(p-NFκB-p65)(P<0.01),and down-regulation of the jejunal villus height,the protein and mRNA abundances of zonula occludens-1(ZO-1)and occludin(OCC;P<0.05)in jejunal mucosa.Dietary YH supplementation decreased the impaired effects of ACTH,cortisol,HP,IL-1βand diamine oxidase in serum(P<0.05).Moreover,YH supplementation also up-regulated the jejunal villus height,protein and mRNA abundances of ZO-1 and OCC(P<0.05),down-regulated the mRNA expressions of TNF-αand IL-1βand the protein abundances of TNF-α,IL-1β,TLR4 and p-NFκB-p65 in jejunal mucosa in LPS-challenged pigs(P<0.01).Conclusion Yeast hydrolysate could attenuate inflammatory response and intestinal barrier injury in weaned piglets challenged with LPS,which was associated with the inhibition of TLR4/NF-κB signaling pathway activation.

Effect of different dietary protein levels and amino acids supplementation patterns on growth performance, carcass characteristics and nitrogen excretion in growing-finishing pigs

Background:This study was conducted to determine the effects of different dietary protein levels and amino acids supplementation patterns in low protein diets on the growth performance,carcass characteristics and nitrogen excretion in growing-finishing pigs.Forty-two barrows(25.00±0.39 kg)were randomly assigned to 7 diets.Diet 1:the high crude protein diet with balanced for 10 essential amino acids(EAAs).Diet 2:the medium crude protein diet with 2%(approx)decreased protein level of Diet 1 and balanced 10 EAAs.Diet 3:the low crude protein diet with 4%decreased protein level of Diet 1 and balanced 10 EAAs.The protein levels of Diet 4,5,6 and 7 were the same as that of Diet 3.Diet 4 was only balanced for lysine(Lys),methionine(Met),threonine(Thr)and tryptophan(Trp);Diet 5 and 6 were further supplemented with extra isoleucine(Ile)or valine(Val),respectively;Diet 7 was further supplemented with extra Ile+Val.Results:Over the 112 days trial,the reduction of dietary protein by 2%or 4%with balanced10 EAAs significantly decreased nitrogen excretion(P<0.05),but had no effects on growth performance and carcass characteristics(P>0.05).In low protein diet,Val supplementation significantly increased body weight gain at 25–50 kg phase(P<0.05),while Ile supplementation at 75–100 kg phase and 100–125 kg phase significantly reduced the ratio of feed to gain(P<0.05).No effect of different dietary protein levels and amino acids supplementation patterns in low protein diets on carcass characteristics was observed(P>0.05).The total N excretion of pigs supplemented with only Lys,Met,Thr and Trp was numerically higher than that of pigs fed with extra Ile,or Val,or Ile+Val diets.Conclusion:In low protein diet,Val is more required than Ile in the early growing phage(25–50 kg),while Ile becomes more required in the late growing and finishing phage(75–125 kg).

The toxicological effect of dietary excess of saccharicterpenin,the extract of camellia seed meal,in piglets

Recently, saccharicterpenin extracted from Camellia oleifera seed meal has become a widely used feed supplement in animal husbandry. In order to assess its safety, this study was designed to investigate the toxicity and histopathological effects of saccharicterpenin on piglets. One hundred-fifty weaned pigs((Yorkshire×Landrace)×Duroc), 75 males and 75 females with body weight(BW) of(7.35±0.29) kg, were randomly allotted to groups receiving diets supplemented with 0, 500, 1 000, 2 500 or 5 000 mg kg^–1 saccharicterpenin for 70 d. The diet with 500 mg kg^–1 saccharicterpenin supplementation improved liver glutathione peroxidase(GSH-Px) activity, and the diet with 1 000 mg kg^–1 saccharicterpenin supplementation improved liver glutathione S-transferase(GSH-S) activity in piglets on d 70(P<0.05). At 2 500 mg kg^–1, saccharicterpenin in the diets reduced average daily feed intake(ADFI) and average daily gain(ADG) of piglets from d 1 to 35, damaged the cardiac tissue and liver on d 35, and decreased white blood cell counts(WBC), activities of catalase(CAT) and GSH-Px, and concentrations of glucose(GLU) and urea nitrogen(BUN) in the blood of piglets on d 70(P<0.05). In addition, diets with 5 000 mg kg^–1 saccharicterpenin supplementation reduced ADFI, ADG and increased diarrhea rates of piglets from d 36 to 70, and decreased hemoglobin(HGB) concentration and activity of CAT in the blood of piglets on d 70(P<0.05). Moreover, at a rate of 5 000 mg kg^–1, saccharicterpenin supplementation increased pancreas index on d 35 and hepatic index on d 70, and damaged cardiac tissue, liver and spleen during the whole experimental period(P<0.05). These results suggested that dietary 500 mg kg^–1 saccharicterpenin supplementation had beneficial effects on piglets, but excessive supplementation(2 500 or 5 000 mg kg^–1) of saccharicterpenin in the diets could lead to growth retardation, hematological abnormalities and organ injuries.

Lentinan administration alleviates diarrhea of rotavirus-infected weaned pigs via regulating intestinal immunity

Background:Lentinan(LNT)may regulate many important physiological functions of human and animals.This study aimed to verify whether LNT administration could relieve diarrhea via improving gut immunity in rotavirus(RV)-challenged weaned pigs.Methods:Twenty-eight weaned pigs were randomly fed 2 diets containing 0 or 84 mg/kg LNT product for 19 d(n=14).RV infection was executed on d 15.After extracting polysaccharides from LNT product,its major monosaccharides were analyzed.Then,LNT polysaccharide was used to administrate RV-infected IPEC-J2 cells.Results:Dietary LNT supplementation supported normal function of piglets even when infected with RV,as reflected by reduced growth performance loss and diarrhea prevalence,and maintained gut immunity(P<0.05).The polysaccharide was isolated from LNT product,which molecular weight was 5303 Da,and major monosaccharides included glucose,arabinose and galactose.In RV-infected IPEC-J2 cells,this polysaccharide significantly increased cell viability(P<0.05),and significantly increased anti-virus immunity via regulating pattern recognition receptors and host defense peptides(P<0.05).Conclusion:Those results suggest that LNT administration increases the piglets’resistance to RV-induced stress,likely by supporting intestinal immunity.

The effect of dietary tryptophan levels on oxidative stress of liver induced by diquat in weaned piglets

Oxidative stress can induce abnormal tryptophan metabolism. The present study was mainly conducted to determine the effect of dietary tryptophan levels on oxidative stress in the liver of weaned pigs challenged by diquat. A total of 36 PIC piglets weaned at 21 days of age were randomly allotted to 1 of 3 diets containing dietary tryptophan levels of 0.18, 0.30, and 0A5% for 14 d. On day 8, the piglets were injected intraperitoneally with sterile 0.9% NaCI solution or diquat （10 mg/kg body weight）. During the first 7 d of trial, increasing dietary tryptophan levels enhanced average daily gain （P = 0.09） and average daily feed intake （P = 0.08）, and decreased the feed efficiency （P 〈 0.05） of piglets. The growth performance was decreased by diquat injection （P 〈 0.05）. Diquat injection also decreased the activities of the superoxide dismutase （SOD） and glutathione peroxidase （GPx） in the plasma and liver （P 〈 0.05）, increased plasma malondialdehyde （MDA） （P 〈 0.05） and urea nitrogen （P 〈 0.05） concentrations, and enhanced MDA concentration （P = 0.09） and tryptophan 2,3-dioxygenase （TDO） activity （P = 0.07） in liver of piglets. Increasing dietary tryptophan levels could attenuate the effects of diquat injection on the MDA （P = 0.06） concentration and the activities of SOD （P = 0.09） and GPx （P = 0.05） of the liver, and plasma urea nitrogen （P = 0.06） concentration in the piglet. There was a synergistic role for increasing TDO activity in the liver between dietary tryptophan levels and diquat injection （P 〈 0.05）. These results suggest that increasing dietary tryptophan levels could attenuate the oxidative stress of the liver in weaned piglets intraperitoneally injected with diquat via enhancing the antioxidant capacity.

Short-chain fatty acids can improve lipid and glucose metabolism independently of the pig gut microbiota

Background:Previous studies have shown that exogenous short-chain fatty acids(SCFAs)introduction attenuated the body fat deposition in conventional mice and pigs.However,limited studies have evaluated the effects of exogenously introduced SCFAs on the lipid and glucose metabolism independently of the gut microbiota.This study was to investigate the effects of exogenous introduction of SCFAs on the lipid and glucose metabolism in a germ-free(GF)pig model.Methods:Twelve hysterectomy-derived newborn pigs were reared in six sterile isolators.All pigs were hand-fed with sterile milk powder for 21 d,then the sterile feed was introduced to pigs for another 21 d.In the second 21-d period,six pigs were orally administrated with 25 mL/kg sterile saline per day and considered as the GF group,while the other six pigs were orally administrated with 25 mL/kg SCFAs mixture(acetic,propionic,and butyric acids,45,15,and 11 mmol/L,respectively)per day and regarded as FA group.Results:Orally administrated with SCFAs tended to increase the adiponectin concentration in serum,enhance the CPT-1 activity in longissimus dorsi,and upregulate the ANGPTL4 mRNA expression level in colon(P<0.10).Meanwhile,the mRNA abundances of ACC,FAS,and SREBP-1C in liver and CD36 in longissimus dorsi of the FA group were decreased(P<0.05)compared with those in the GF group.Besides,the mRNA expression of PGC-1αin liver and LPL in longissimus dorsi tended to(P<0.10)upregulate and downregulate respectively in the FA group.Moreover,oral administration of SCFAs tended to increase the protein level of GPR43(P<0.10)and decrease the protein level of ACC(P<0.10)in liver.Also,oral administration of SCFAs upregulated the p-AMPK/AMPK ratio and the mRNA expressions of GLUT-2 and GYS2 in liver(P<0.05).In addition,the metabolic pathway associated with the biosynthesis of unsaturated fatty acids was most significantly promoted(P<0.05)by oral administration of SCFAs.Conclusions:Exogenous introduction of SCFAs might attenuate the fat deposition and to some extent improve the glucose control in the pig model,which occurred independently of the gut microbiota.

Effect of sialyllactose on growth performance and intestinal epithelium functions in weaned pigs challenged by enterotoxigenic Escherichia Coli

Background:Sialyllactose(SL)is one of the most abundant oligosaccharides present in porcine breast milk.However,little is known about its effect on growth performance and intestinal health in weaned pigs.This study was conducted to explore the protective effect of SL on intestinal epithelium in weaned pigs upon enterotoxigenic Escherichia coli(ETEC)challenge.Methods:Thirty-two pigs were randomly divided into four treatments.Pigs fed with a basal diet or basal diet containing SL(5.0 g/kg)were orally infused with ETEC or culture medium.Results:SL supplementation elevated the average daily gain(ADG)and feed efficiency in the ETEC-challenged pigs(P<0.05).SL also improved the digestibilities of dry matter(DM),gross energy(GE),and ash in non-challenged pigs(P<0.05).Moreover,SL not only elevated serum concentrations of immunoglobulins(IgA,IgG,and IgM),but also significantly decreased the serum concentrations of inflammatory cytokines(TNF-α,IL-1β,and IL-6)upon ETEC challenge(P<0.05).Interestingly,SL increased the villus height,the ratio of villus height to crypt depth(V:C),and the activities of mucosal sucrase and maltase in the jejunum and ileum(P<0.05).SL also elevated the concentrations of microbial metabolites(e.g.acetic acid,propanoic acid,and butyric acid)and the abundance of Lactobacillus,Bifidobacterium,and Bacillus in the cecum(P<0.05).Importantly,SL significantly elevated the expression levels of jejunal zonula occludins-1(ZO-1),occluding,and fatty acid transport protein-4(FATP4)in the ETEC-challenged pigs(P<0.05).Conclusions:SL can alleviate inflammation and intestinal injury in weaned pigs upon ETEC challenge,which was associated with suppressed secretion of inflammatory cytokines and elevated serum immunoglobulins,as well as improved intestinal epithelium functions and microbiota.

Novel metabolic and physiological functions of branched chain amino acids: a review

It is widely known that branched chain amino acids（BCAA） are not only elementary components for building muscle tissue but also participate in increasing protein synthesis in animals and humans. BCAA（isoleucine, leucine and valine） regulate many key signaling pathways, the most classic of which is the activation of the m TOR signaling pathway. This signaling pathway connects many diverse physiological and metabolic roles. Recent years have witnessed many striking developments in determining the novel functions of BCAA including：（1） Insufficient or excessive levels of BCAA in the diet enhances lipolysis.（2） BCAA, especially isoleucine, play a major role in enhancing glucose consumption and utilization by up-regulating intestinal and muscular glucose transporters.（3）Supplementation of leucine in the diet enhances meat quality in finishing pigs.（4） BCAA are beneficial for mammary health, milk quality and embryo growth.（5） BCAA enhance intestinal development, intestinal amino acid transportation and mucin production.（6） BCAA participate in up-regulating innate and adaptive immune responses.In addition, abnormally elevated BCAA levels in the blood（decreased BCAA catabolism） are a good biomarker for the early detection of obesity, diabetes and other metabolic diseases. This review will provide some insights into these novel metabolic and physiological functions of BCAA.

Beneficial effects of a decreased meal frequency on nutrient utilization,secretion of luteinizing hormones and ovarian follicular development in gilts

Background:Replacement gilts are typically fed ad libitum,whereas emerging evidence from human and rodent studies has revealed that time-restricted access to food has health benefits.The objective of this study was to investigate the effect of meal frequency on the metabolic status and ovarian follicular development in gilts.Methods:A total of 36 gilts(Landrace×Yorkshire)with an age of 150±3 d and a body weight of 77.6±3.8 kg were randomly allocated into one of three groups(n=12 in each group),and based on the group allocation,the gilts were fed at a frequency of one meal(T1),two meals(T2),or six meals per day(T6)for 14 consecutive weeks.The effects of the meal frequency on growth preference,nutrient utilization,short-chain fatty acid production by gut microbial,the post-meal dynamics in the metabolic status,reproductive hormone secretions,and ovarian follicular development in the gilts were measured.Results:The gilts in the T1 group presented a higher average daily gain(+48 g/d,P<0.05)and a higher body weight(+4.9 kg,P<0.05)than those in the T6 group.The meal frequency had no effect on the apparent digestibility of dry matter,crude protein,ether extract,ash,and gross energy,with the exception that the T1 gilts exhibited a greater NDF digestibility than the T6 gilts(P<0.05).The nitrogen balance analysis revealed that the T1 gilts presented decreased urine excretion of nitrogen(−8.17 g/d,P<0.05)and higher nitrogen retention(+9.81 g/d,P<0.05),and thus exhibited higher nitrogen utilization than the T6 gilts.The time-course dynamics of glucose,α-amino nitrogen,urea,lactate,and insulin levels in serum revealed that the T1 group exhibited higher utilization of nutrients after a meal than the T2 or T6 gilts.The T1 gilts also had a higher acetate content and SCFAs in feces than the T6 gilts(P<0.05).The age,body weight and backfat thickness of the gilts at first estrous expression were not affected by the meal frequency,but the gilts in the T1 group had higher levels of serum luteinizing hormone on the 18th day of the 3rd estrus cycle and 17β-estradiol,a larger number of growing follicles and corpora lutea,and higher mRNA expression levels of genes related to follicular development on the 19th day of the 3rd estrus cycle.Conclusions:The current findings revealed the benefits of a lower meal frequency equal feed intake on nutrient utilization and reproductive function in replacement gilts,and thus provide new insights into the nutritional strategy for replacement gilts,and the dietary pattern for other mammals,such as humans.

Cytoprotection and Cytothetic Effects of GLP-2 on Enterocytes from a Weaned Piglet Injured by β-conglycinin in Vitro

The experiment was conducted to study the effects and possible mechanism of GLP-2 on proliferation,metabolism and apoptosis of cultured enterocytes from a 28-d weaned piglet injured by exposure to β-conglycinin.A cell damage model was established to investigate cell proliferation, metabolism and apoptosis by exposing primary cell cultures of intestinal epithelial cells(IEC) to 1.2 and 2.4 mg/mL β-conglycinin.A 2×3 factorial experiment was then used to study the effect of different GLP-2 concentrations of(1×10-9,1×10-8 and 1×10-7mol/L),in combination with the two concentrations ofβ-conglycinin.Cells exposed to the allergenβ-conglycinin had decreased(P【0.05) MTT OD;decreased (P【0.01) protein retention and total protein content of cells;increased(P【0.01) LDH and caspase-3 activities and decreased(P【0.05) Na+,K+-ATPase activity.When GLP-2 was used in combination withβ-conglycinin,MTT OD,protein retention,total protein content and Na+,K+-ATPase activity significantly increased(P【0.05);LDH activity gradually decreased(P【0.05 or P【0.01) and Caspase-3 activity significantly decreased(P【0.01) with increasing concentrations of GLP-2.The results indicated thatβ-conglycinin had adverse effects on proliferation and integrity of IEC in vitro.GLP-2 relieved or prevented the adverse effects ofβ-conglycinin on proliferation and integrity of IEC by regulating Na+,K+- ATPase and Caspase-3 activities,and consequently affecting cell metabolism.

25-hydroxycholecalciferol reverses heat induced alterations in bone quality in finisher broilers associated with effects on intestinal integrity and inflammation

Background:Alterations in ambient temperature have been associated with multiple detrimental effects on broilers such as intestinal barrier disruption and dysbiosis resulting in systemic inflammation.Inflammation and 25-hydroxycholecalciferol(25-OH-D_(3))have shown to play a negative and positive role,respectively,in the regulation of bone mass.Hence the potential of 25-OH-D_(3)in alleviating heat induced bone alterations and its mechanisms was studied.Results:Heat stress(HS)directly induced a decrease in tibia material properties and bone mass,as demonstrated by lower mineral content,and HS caused a notable increase in intestinal permeability.Treatment with dietary 25-OH-D_(3)reversed the HS-induced bone loss and barrier leak.Broilers suffering from HS exhibited dysbiosis and increased expression of inflammatory cytokines in the ileum and bone marrow,as well as increased osteoclast number and activity.The changes were prevented by dietary 25-OH-D_(3)administration.Specifically,dietary 25-OH-D_(3)addition decreased abundance of B-and T-cells in blood,and the expression of inflammatory cytokines,especially TNF-α,in both the ileum and bone marrow,but did not alter the diversity and population or composition of major bacterial phyla.With regard to bone remodeling,dietary 25-OH-D_(3)supplementation was linked to a decrease in serum C-terminal cross-linked telopeptide of type I collagen reflecting bone resorption and a concomitant decrement in osteoclast-specific marker genes expression(e.g.cathepsin K),whereas it did not apparently change serum bone formation markers during HS.Conclusions:These data underscore the damage of HS to intestinal integrity and bone health,as well as that dietary 25-OH-D_(3)supplementation was identified as a potential therapy for preventing these adverse effects.

The role of vitamin E in polyunsaturated fatty acid synthesis and alleviating endoplasmic reticulum stress in sub-adult grass carp(Ctenopharyngodon idella)

Vitamin E(VE)is an essential lipid-soluble vitamin that improves the fish flesh quality.However,the underlying molecular mechanisms remain unclear.This study aimed to investigate the effects of VE on growth performance and flesh quality in sub-adult grass carp(Ctenopharyngodon idella).A total of 450fish(713.53±1.50 g)were randomly divided into six treatment groups(three replicates per treatment)and fed for nine weeks with different experimental diets(dietary lipid 47.8 g/kg)that contained different levels of VE(5.44,52.07,96.85,141.71,185.66,and 230.12 mg/kg diet,supplemented as dl-a-tocopherol acetate).Notably,the treatment groups that were fed with dietary VE ranging from 52.07 to230.12 mg/kg diet showed improvement in the percent weight gain,special growth rate,and feed efficiency of grass carp.Moreover,the treatment groups supplemented with dietary VE level of 141.71,185.66,and 230.12 mg/kg diet showed enhancement in crude protein,lipid,and a-tocopherol contents in the muscle,and the dietary levels of VE ranging from 52.07 to 141.71 mg/kg diet improved muscle pH24h and shear force but reduced muscle cooking loss in grass carp.Furthermore,appropriate levels of VE(52.07 to 96.85 mg/kg diet)increased the muscle polyunsaturated fatty acid content in grass carp.Dietary VE also increased the mRNA levels of fatty acid synthesis-related genes,including fas,scd-1,fad,elovl,srebp1,pparγ,and lxra,and up-regulated the expression of SREBP-1 protein.However,dietary VE decreased the expression of fatty acid decomposition-related genes,including hsl,cpt1,acox1,and ppara,and endoplasmic reticulum stress-related genes,including perk,ire1,atf6,eif2a,atf4,xbp1,chop,and grp78,and down-regulated the expression of p-PERK,p-IRE1,ATF6,and GRP78 proteins.In conclusion,dietary VE increased muscle fatty acid synthesis,which may be partly associated with the alleviation of endoplasmic reticulum stress,and ultimately improves fish flesh quality.Moreover,the VE requirements for sub-adult grass carp(713.53 to 1590.40 g)were estimated to be 124.9 and 122.73 mg/kg diet based on percentage weight gain and muscle shear force,respectively.

Secondary bile acids are associated with body lipid accumulation in obese pigs

The aim of this study was to investigate the reasons for the differences in lipid accumulation between lean and obese pigs.The bile acids with varying levels within two types of pigs were found and then in vitro experiments were conducted to identify whether these bile acids can directly affect lipid accumulation.Fourteen pigs,including seven lean and seven obese pigs with body weights of approximately 80 kg,were fed the same diet at an amount approximately equivalent to 3%of their respective body weights daily for 42 d.In vitro,3T3-L1 preadipocytes were cultured in medium with high glucose levels and were differentiated into mature adipocytes using differentiation medium.Then,bile acids were added to mature adipocytes for 4 d.The results showed that there was a difference in body lipids levels and gut microbiota composition between obese and lean pigs(P<0.05).According to the results of gut microbial function prediction,the bile acid biosynthesis in colonic digesta of obese pigs were different from that in lean pig.Sixty-five bile acids were further screened by metabolomics,of which 4 were upregulated(P<0.05)and 2 were downregulated(P<0.05)in obese pigs compared to lean pigs.The results of the correlation analysis demonstrated that chenodeoxycholic acid-3-β-D-glucuronide(CDCA-3Gln)andω-muricholic acid(ω-MCA)had a negative correlation with abdominal fat weight and abdominal fat rate,while isoallolithocholic acid(IALCA)was positively associated with crude fat in the liver and abdominal fat rate.There was a positive correlation between loin muscle area and CDCA-3Gln andω-MCA(P<0.05),however,IALCA and 3-oxodeoxycholic acid(3-oxo-DCA)were negatively associated with loin eye muscle area(P<0.05).Isoallolithocholic acid increased the gene expression of peroxisome proliferator-activated receptor gamma(PPARG)and the number of lipid droplets(P<0.05),promoting the lipid storage when IALCA was added to 3T3-L1 mature adipocytes in vitro.In conclusion,the concentration of bile acids,especially gut microbiota related-secondary bile acids,in obese pigs was different from that in lean pigs,which may contribute to lipid accumulation within obese pigs.

Establishing the link between D-mannose and juvenile grass carp(Ctenopharyngodon idella):Improved growth and intestinal structure associated with endoplasmic reticulum stress,mitophagy,and apical junctional complexes

D-mannose,essential for protein glycosylation,has been reported to have immunomodulatory effects and to maintain intestinal flora homeostasis.In addition to evaluating growth performance,we examined the impact of D-mannose on the structure of epithelial cells and apical junction complexes in the animal intestine.All 1800 grass carp(16.20±0.01 g)were randomly divided into six treatments with six replicates of 50 fish each and fed with six different levels of D-mannose(0.52,1.75,3.02,4.28,5.50 and 6.78 g/kg diet)for 70 d.The study revealed that D-mannose increased feed intake(P 0.05).D-mannose supplementation at 1.75 g/kg increased crude protein content in fish and lipid production value(P<0.05).D-mannose supplementation at 4.28 g/kg increased intestinal length,intestinal weight and fold height of grass carp compared to the control group(P<0.05).This improvement may be attributed to the phosphomannose isomerase(PMI)-mediated enhancement of glycolysis.This study found that D-mannose supplementation at 4.28 or 3.02 g/kg reduced serum diamine oxidase activity or D-lactate content(P<0.05)and improved cellular and intercellular structures for the first time.The improvement of cellular redox homeostasis involves alleviating endoplasmic reticulum(ER)stress through the inositol-requiring enzyme 1(IRE1),RNA-dependent protein kinase-like ER kinase(PERK),and activating transcription factor 6(ATF6)signaling pathways.The alleviation of ER stress may be linked to the phosphomannomutase(PMM)-mediated enhancement of protein glycosylation.In addition,ubiquitin-dependent[PTEN-induced putative kinase 1(PINK1)/Parkin]and ubiquitin-independent[BCL2-interacting protein 3-like(BNIP3L),BCL2-interacting protein 3(BNIP3),and FUN14 domain containing 1(FUNDC1)]mitophagy may play a role in maintaining cellular redox homeostasis.The enhancement of intercellular structures includes enhancing tight junction and adherent junction structures,which may be closely associated with the small Rho GTPase protein(RhoA)/the Rho-associated protein kinase(ROCK)signaling pathway.In conclusion,D-mannose improved intestinal cellular redox homeostasis associated with ER stress and mitophagy pathways,and enhanced intercellular structures related to tight junctions and adherent junctions.Furthermore,quadratic regression analysis of the PWG and intestinal reactive oxygen species content indicated that the optimal addition level of D-mannose for juvenile grass carp was 4.61 and 4.59 g/kg,respectively.

Dietary curcumin alleviates intestinal damage induced by ochratoxin A in juvenile grass carp(Ctenopharyngodon idella):Necroptosis and inflammatory responses

Ochratoxin A(OTA)is one of the most common pollutants in aquatic feed.As a first line of defense,intestinal barriers could be utilized against OTA in order to prevent disorders.Natural product supplementation is one of the most popular strategies to alleviate toxicity induced by mycotoxins,but there is a lack of knowledge about how it functions in the teleost intestine.In this study,720 juvenile grass carp of about 11 g were selected and four treatment groups(control group,OTA group,curcumin[Cur]group,and OTA+Cur group)were set up to conduct a 60-day growth test.After the test,the growth performance and intestinal health related indexes of grass carp were investigated.The addition of dietary Cur could have the following main results:(1)inhibit absorption and promote efflux transporters mRNA expression,reducing the residuals of OTA,(2)decrease oxidative stress by reducing oxidative damage and enhancing the expression of antioxidant enzymes,(3)promote mitochondrial fusion proteins to inhibit the expression of mitotic proteins and mitochondrial autophagy proteins and enhance mitochondrial function,(4)reduce necroptosis-related gene expression through inhibiting the tumor necrotic factor receptor-interacting protein kinase/mixed lineage kinase domain-like pathway,(5)reduce the expression of pro-inflammatory factors by inhibiting the Toll-like receptor 4/nuclear factor-κB signaling pathway to alleviate the intestinal inflammatory response.In summary,the results suggested that Cur could alleviate OTA-induced intestinal damage by enhancing antioxidant capacity and mitochondrial function as well as reducing necroptosis and inflammation in the grass carp intestine.This study provided a theoretical basis and production implications for dietary Cur that could improve growth performance and alleviate the intestinal damage induced by OTA in fish.

Dietary copper improves intestinal structural integrity in juvenile grass carp(Ctenopharyngodon idella)probably related to its increased intestinal antioxidant capacity and apical junction complex

This research evaluated the effects of copper(Cu)on intestinal antioxidant capacity and apical junctional complex(AJC)in juvenile grass carp.A total of 1080 healthy juvenile grass carp(11.16±0.01 g)were fed six diets including different dosages of Cu,namely 0,2,4,6,8 mg/kg(Cu citrate[CuCit]as Cu source)and 3 mg/kg(CuSO_(4)·5H_(2)O as Cu source).The trial lasted for 9 weeks.The findings revealed that dietary optimal Cu supplementation(2.2 to 4.1 mg/kg)promoted intestinal growth,including intestinal length,intestinal length index,intestinal weight,and intestinal somatic index(P>0.05).On the other hand,optimal Cu increased intestinal tight junction(TJ)proteins(except for claudin 15b)and adherens junction(AJ)proteins(E-cadherin,α-catenin,β-catenin,nectin and afadin)mRNA levels(P<0.05),which could be connected to the signaling pathway formed by the Ras homolog gene family,member A(RhoA),Rho-associated kinase(ROCK),and myosin light chain kinase(MLCK).Finally,based on serum indicator D-lactate and intestinal oxidative damage index(ROS),Cu requirement(CuCit as Cu source)for juvenile grass carp from initial weight to final weight(from 11 to 173 g)was determined to be 4.14 and 4.12 mg/kg diet,respectively.This work may provide a theoretical foundation for identifying putative Cu regulation pathways on fish intestinal health.

A new insight on copper:Promotion of collagen synthesis and myofiber growth and development in juvenile grass carp(Ctenopharyngodon idella)

Copper(Cu)is a trace element,essential for fish growth.In the current study,in addition to growth performance,we first explored the effects of Cu on collagen synthesis and myofiber growth and development in juvenile grass carp(Ctenopharyngodon idella).A total of 1080 fish(11.16±0.01 g)were randomly divided into 6 treatments(3 replicates per treatment)to receive five doses of organic Cu which were Cu citrate(CuCit)at 0.99(basal diet),2.19,4.06,6.15,and 8.07 mg/kg,and one dose o inorganic Cu(CuSO_(4)·5H_(2)O at 3.15 mg/kg),for 9 weeks.The results showed appropriate Cu level(4.06 mg kg)enhanced growth performance,improved nutritional Cu status,and downregulated Cu-transporting ATPase 1 mRNA levels in the hepatopancreas,intestine,and muscle of juvenile grass carp.Meanwhile collagen content in fish muscle was increased after Cu intake,which was probably due to the following pathways:(1)activating CTGF/TGF-β1/Smads signaling pathway to regulate collagen transcription;(2upregulating of La ribonucleoprotein domain family 6(LARP6)mRNA levels to regulate translation initiation;(3)increasing proline hydroxylase,lysine hydroxylase,and lysine oxidase activities to regulate posttranslational modifications.In addition,optimal Cu group increased myofiber diameters and the frequency of myofibers with diameter>50μm,which might be associated with upregulation of cyclin B cyclin D,cyclin E,proliferating cell nuclear antigen,myogenic determining factor(MyoD),myogenic factor 5,myogenin(MyoG),myogenic regulatory factor 4 and myosin heavy chain(MyHC)and down regulation of myostatin mRNA levels,increasing protein levels of MyoD,MyoG and MyHC in fish muscle Finally,based on percentage weight gain(PWG),serum ceruloplasmin(Cp)activity and collagen conten in fish muscle,Cu requirements were determined as 4.74,4.37 and 4.62 mg/kg diet(CuCit as Cu source of juvenile grass carp,respectively.Based on PWG and Cp activity,compared to CuSO_(4)·5H_(2)O,the efficacy of CuCit were 131.80%and 115.38%,respectively.Our findings provide new insights into Cu supple mentation to promote muscle growth in fish,and help improve the overall productivity of aquaculture.

Effect of fermented rapeseed meal on growth performance,nutrient digestibility,and intestinal health in growing pigs

To explore the effects of fermented rapeseed meal(FRSM)on growth performance and intestinal health,a total of 30 growing pigs were randomly allotted to three treatments consisting of corn-soybean meal diet(CSD),rapeseed meal diet(RSD),and fermented rapeseed meal diet(FRSD).Results showed that compared with RSD,FRSD feeding increased the average daily gain and final body weight in pigs(P<0.01).Compared with RSD feeding,FRSD feeding elevated the apparent digestibility of crude protein,acid detergent fiber,and ether extract in pigs(P<0.01).Moreover,the FRSD group exhibited greater apparent ileal digestibility of His,Thr,Lys,and Ser than the RSD group(P<0.01).The digestible energy,metabolic energy,and nitrogen utilization were higher in the FRSD and CSD groups than in the RSD group(P<0.01).As compared to the RSD,FRSD feeding decreased the serum concentration of leptin but significantly increased the concentrations of immunoglobulin(Ig)A,IgG,ghrelin,and enzyme activities of amylase,lipase,and trypsin in the pancreas(P<0.05).Interestingly,the villus height,the ratio of villus height to crypt depth,and the activities of brush border enzymes(e.g.,maltase and sucrase)in the small intestine were higher in the CSD and FRSD groups than in the RSD group(P<0.05).As compared to the RSD,the FRSD feeding not only increased the expression level of the occludin in the small intestinal epithelium(P<0.05)but also elevated the expression levels of claudin-1,MUC1,and PepT1 genes in the duodenum,and elevated the expression levels of SGLT1 and CAT1 genes in the jejunum(P<0.05).Importantly,FRSD feeding significantly decreased the abundance of Escherichia coli,but increased the abundance of Lactobacillus and the content of butyrate in the cecum and colon(P<0.05).These results indicated that compared with rapeseed meal,fermented rapeseed meal exhibited a positive effect on improving the growth performance and intestinal health in growing pigs,and the results may also help develop novel protein sources for animal nutrition and the feed industry.