Dietary supplementation of Macleaya cordata extract and Bacillus in combination improve laying performance by regulating reproductive hormones,intestinal microbiota and barrier function of laying hens

Background:This study aimed to investigate whether the combination of Macleaya cordata extract(MCE)and Bacil-lus could improve the laying performance and health of laying hens better.Methods:A total of 36029-week-old Jingbai laying hens were randomly divided into 4 treatments:control group(basal diet),MCE group(basal diet+MCE),Probiotics Bacillus Compound(PBC)group(basal diet+compound Bacil-lus),MCE+PBC group(basal diet+MCE+compound Bacillus).The feeding experiment lasted for 42 d.Results:The results showed that the laying rate and the average daily egg mass in the MCE+PBC group were significantly higher than those in the control group(P<0.05)and better than the MCE and PBC group.Combina-tion of MCE and Bacillus significantly increased the content of follicle-stimulating hormone(FSH)in the serum and up-regulated the expression of related hormone receptor gene(estrogen receptor-β,FSHR and luteinizing hormone/choriogonadotropin receptor)in the ovary of laying hens(P<0.05).In the MCE+PBC group,the mRNA expressions of zonula occluden-1,Occludin and mucin-2 in jejunum was increased and the intestinal epithelial barrier detected by transmission electron microscopy was enhanced compared with the control group(P<0.05).In addition,compared with the control group,combination of MCE and Bacillus significantly increased the total antioxidant capacity and catalase activity(P<0.05),and down-regulated the mRNA expressions of inflammation-related genes(interleukin-1βand tumor necrosis factor-α)as well as apoptosis-related genes(Caspase 3,Caspase 8 and P53)(P<0.05).The concen-tration of acetic acid and butyric acid in the cecum content of laying hens in the MCE+PBC group was significantly increased compared with the control group(P<0.05).Conclusions:Collectively,dietary supplementation of 600μg/kg MCE and 5×108 CFU/kg compound Bacillus can improve laying performance by improving microbiota to enhance antioxidant capacity and intestinal barrier,regulate reproductive hormones and the concentration of cecal short-chain fatty acids of laying hens,and the combined effect of MCE and Bacillus is better than that of single supplementation.

Preparation, characterization, antimicrobial and cytotoxicity studies of copper/zincloaded montmorillonite

Background: A series of modified montmorillonites(Mt) including zinc-loaded Mt(Zn-Mt), copper-loaded Mt(Cu-Mt), copper/zinc-loaded Mt with different Cu/Zn ratio(Cu/Zn-Mt-1, Cu/Zn-Mt-2, Cu/Zn-Mt-3) were prepared by an ion-exchange reaction, and characterized using X-ray diffraction(XRD), fourier transformed infrared spectroscopy(FTIR) and transmission electron microscopy(TEM). The specific surface areas, antimicrobial activity and cytotoxicity of the modified Mt were investigated.Results: In the modified Mt, hydrated Cu ions and Zn ions were exchanged in the interlayer space of Mt and the particles were irregular shapes. The results showed that Cu/Zn-Mt enhanced antibacterial and antifungal activity compared with Zn-Mt and Cu-Mt possibly due to the synergistic effect between Cu and Zn. Among the Cu/Zn-Mt with different Cu/Zn raitos, Cu/Zn-Mt with a Cu/Zn ratio of 0.98 or 0.51 showed higher antimicrobial activity against gram-negative bacteria(Escherichia coli), gram-positive bacteria(Staphylococcus aureus), fungi(Candida albicans).Moreover, the antimicrobial activity of Cu/Zn-Mt was correlated with its specific surface area. Cytotoxicity studies on IPEC-J2 cell showed a slight cytotoxicity of Cu/Zn-Mt.Conclusions: The current data provide clear evidence that in terms of its antimicrobial activity and relatively low toxicity, the Cu/Zn-Mt holds great promise for applications in animal husbandry.

Comprehensive evaluation of the metabolic effects of porcine CRTC3 overexpression on subcutaneous adipocytes with metabolomic and transcriptomic analyses

Background:Meat quality is largely driven by fat deposition,which is regulated by several genes and signaling pathways.The cyclic adenosine monophosphate(cAMP)-regulated transcriptional coactivator 3(CRTC3)is a coactivator of cAMP response element binding protein(CREB)that mediates the function of protein kinase A(PKA)signaling pathway and is involved in various biological processes including lipid and energy metabolism.However,the effects of CRTC3 on the metabolome and transcriptome of porcine subcutaneous adipocytes have not been studied yet.Here,we tested whether porcine CRTC3 expression would be related to fat deposition in Heigai pigs(a local fatty breed in China)and Duroc×Landrace×Yorkshire(DLY,a lean breed)pigs in vivo.The effects of adenovirus-induced CRTC3 overexpression on the metabolomic and transcriptomic profiles of subcutaneous adipocytes were also determined in vitro by performing mass spectrometry-based metabolomics combined with RNA sequencing(RNA-seq).Results:Porcine CRTC3 expression is associated with fat deposition in vivo.In addition,CRTC3 overexpression increased lipid accumulation and the expression of mature adipocyte-related genes in cultured porcine subcutaneous adipocytes.According to the metabolomic analysis,CRTC3 overexpression induced significant changes in adipocyte lipid,amino acid and nucleotide metabolites in vitro.The RNA-seq analysis suggested that CRTC3 overexpression alters the expression of genes and pathways involved in adipogenesis,fatty acid metabolism and glycerophospholipid metabolism in vitro.Conclusions:We identified significant alterations in the metabolite composition and the expression of genes and pathways involved in lipid metabolism in CRTC3-overexpressing adipocytes.Our results suggest that CRTC3 might play an important regulatory role in lipid metabolism and thus affects lipid accumulation in porcine subcutaneous adipocytes.

Fermented mixed feed alters growth performance,carcass traits,meat quality and muscle fatty acid and amino acid profiles in finishing pigs

This study was conducted to investigate the effects of fermented mixed feed(FMF)on growth performance,carcass traits,meat quality,muscle amino acid and fatty acid composition and mRNA expression levels of genes related to lipid metabolism in finishing pigs.In the present study,144 finishing pigs(Duroc×Berkshire×Jiaxing Black)were randomly allocated to 3 dietary treatments with 4 replicate pens per group and 12 pigs per pen.The dietary treatments included a basal diet(CON),a basal diet+5%FMF and a basal diet+10%FMF.The experiment lasted 38 d after 4 d of acclimation.The results showed that 5%and 10%FMF significantly increased the average daily gain(ADG)of the females but not the males(P<0.05),but FMF supplementation showed no impact on carcass traits.Moreover,10%FMF supplementation increased the meat color45 min and meat color24 h values,while it decreased the shear force relative to CON(P<0.05).In addition,10%FMF significantly increased the contents of flavor amino acids(FAA),total essential AA(EAA),total non-EAA(NEAA)and total AA relative to CON(P<0.05).Furthermore,the diet supplemented with 10%FMF significantly increased the concentration of n-3 polyunsaturated fatty acids(PUFA),n-6 PUFA and total PUFA,and the PUFA to saturated fatty acids ratio(P<0.05),suggesting that FMF supplementation increased meat quality.Moreover,compared with the CON,10%FMF supplementation increased the mRNA expression of lipogenic genes,including CEBPα,PPARγ,SREBP1 and FABP4,and upregulated the expression of unsaturated fatty acid synthesis(ACAA1 and FADS2).Together,our results suggest that 10%FMF dietary supplementation improved the female pigs’growth performance,improved the meat quality and altered the profiles of muscle fatty acids and amino acids in finishing pigs.This study provides a reference for the production of high-quality pork.

Probiotic Paenibacillus polymyxa 10 and Lactobacillus plantarum 16 enhance growth performance of broilers by improving the intestinal health

With the ever-growing strict prohibitions on antibiotic growth promoters(AGP)in animal production,in-feed probiotics are becoming attractive alternatives to antibiotics in the poultry industry.To investigate the effects of Paenibacillus polymyxa 10 and Lactobacillus plantarum 16 on the growth performance and intestinal health of broilers,540 male Cobb 500 broilers of 1 d old were randomly divided into 3 groups with 6 replicates per group and 30 chicks per replicate.Broilers were fed with either a basal diet or basal diets supplemented with 1×108 colony-forming units(CFU)/kg P.polymyxa 10(BSC10)or L.plantarum 16(Lac16)for 42 d.Results showed that Lac16 treatment improved(P<0.05)the growth performance(body weight and feed conversion)of broilers at the starter phase,while BSC10 treatment slightly improved(P>0.05)the growth performance of the starter phase broilers.The increased villus height(P<0.05)at d 14,21 and 42 and villus height to crypt depth ratio(P<0.05)at d 14 and 21 were observed in the ileum of the 2 probiotic groups.Besides,transmission electron microscopy results showed that the 2 probiotics enhanced the intestinal epithelial barrier.Both probiotic treatments up-regulated(P<0.05)the mRNA expression of fatty acid binding protein 1(FABP1)and sodium-dependent glucose transporters-1(SGLT-1)in the ileal mucosa of broilers at d 21.In addition,BSC10 and Lac16 treatments significantly(P<0.05)increased the relative abundance of short-chain fatty acids-producing bacteria,such as Butyricicoccus pullicaecorum,Faecalibacterium prausnitzii,Lachnospira and Cop-rococcu,and significantly(P<0.05)decreased the relative abundance of enteric pathogens(Escherichia coli,Bacteroides fragilis and Shigella sonnei).Furthermore,the 2 probiotic treatments also increased the positive connection among the intestinal microbes and the carbohydrate metabolism-related pathways of the intes-tinal bacteria(P<0.05),with decreasing(P<0.05)nucleotides biosynthesis-related pathways of the intestinal bacteria.Overall,these results suggest that the 2 probiotics,especially Lac16,have a potential beneficial effect on the growth performance and intestinal health of starter phase broilers.

Effects of starch and gelatin encapsulated vitamin A on growth performance,immune status and antioxidant capacity in weaned piglets

To evaluate the effects of gelatin and starch encapsulated vitamin A on growth performance,immune status and antioxidant capacity in weaned piglets,a total of 96 weaned piglets(body weight=9.11±0.03 kg,30-d-old)were randomly allotted to 3 treatments with 4 replications of 8 piglets each.The 3 treatments were control diet(basal diet without addition of vitamin A),gelatin vitamin A diet(basal diet+13,500 IU/kg gelatin encapsulated vitamin A),and starch vitamin A diet(basal diet+13,500 IU/kg starch encapsulated vitamin A),respectively.The results showed that piglets fed starch vitamin A diet had significantly higher final body weight and average daily gain compared to those in control and gelatin vitamin A groups(P<0.05).Gelatin and starch vitamin A supplementation both highly increased serum retinol concentration and immunoglobulin(Ig)M level when compared with the control group(P<0.05).Additionally,serum IgA level and glutathione peroxidase(GSH-Px)activity were significantly increased by gelatin vitamin A diet on d 21 and starch vitamin A diet on d 42,respectively(P<0.05).These results demonstrated that dietary supple mentation of vitamin A could improve immune function and antioxidant capacity in weaned piglets,and starch vitamin A is better than gelatin vitamin A,especially in promoting the growth performance of piglets.

Comparison of the meat quality and fatty acid profile of muscles in finishing Xiangcun Black pigs fed varied dietary energy levels

To study the effects of dietary energy level on the meat quality of different muscles in finishing pigs,400Xiangcun Black pigs(BW=79.55±4.77 kg)were randomly assigned to 5 treatments with varied calculated digestive energy(DE)at 3,050,3,100,3,150,3,200 and 3,250 kcal/kg,respectively.Each treatment had 8replicates with 10 pigs per replicate.Meat quality,amino acid and fatty acid composition were tested in this study.No differences in average daily gain,average daily feed intake or feed-to-gain ratio(P>0.05)were observed among dietary treatments.Glycogen concentrations of longissimus dorsi(LD)muscle in DE3150was higher than those in other groups(P<0.05).The crude fat concentration of biceps femoris(BF)muscle in DE3250 tended to be higher than that in DE3150 and DE3100 groups(P<0.05).Pigs in DE3250 and DE3200 had higher fiber density and smaller cross-sectional area of BF muscle than those in DE3150(P<0.05).Pigs in DE3150 had the highest Cu concentration in LD muscle compared with those in DE3200,DE3250(P<0.05).The C16:1 proportion of LD muscle was lower(P<0.01)and C20:1 was higher(P<0.05)in DE3050 than that in the other dietary treatments.The C18:3n6 and C20:3n6 proportions of BF muscle in DE3150 were higher than those in DE 3050,DE3200 and DE3250(P<0.05).For LD muscle,m RNA expressions of type I and IIa MyHC in group DE3150 were higher than other treatments(P<0.01).The LD muscle in DE3150 expressed higher PPARd than in other groups(P<0.01).Pigs in DE3100 expressed higher FOX1 than in DE3200 and DE3250(P<0.05).Sterol-regulatory element binding proteins(SREBPa)mRNA expression decreased linearly when dietary energy level increased in BF muscle(P<0.01).In conclusion,a200 kcal/kg decrease in digestible energy for 4 consecutive weeks did not affect growth performance of Xiangcun Black pigs.Furthermore,LD and BF muscle respond differently to dietary energy level,and meat quality was improved by the medium energy level during the finishing phase.

Microbiota-gut-brain axis and nutritional strategy under heat stress

Heat stress is a very universal stress event in recent years.Various lines of evidence in the past literatures indicate that gut microbiota composition is susceptible to variable temperature.A varied microbiota is necessary for optimal regulation of host signaling pathways and disrupting microbiota-host homeostasis that induces disease pathology.The microbiota-gut-brain axis involves an interactive mode of communication between the microbes colonizing the gut and brain function.This review summarizes the effects of heat stress on intestinal function and microbiota-gut-brain axis.Heat stress negatively affects intestinal immunity and barrier functions.Microbiota-gut-brain axis is involved in the homeostasis of the gut microbiota,at the same time,heat stress affects the metabolites of microbiota which could alter the function of microbiota-gut-brain axis.We aim to bridge the evidence that the microbiota is adapted to survive and thrive in an extreme environment.Additionally,nutritional strategies for alleviating intestinal heat stress are introduced.

Plasma non-transferrin-bound iron uptake by the small intestine leads to intestinal injury and intestinal flora dysbiosis in an iron overload mouse model and Caco-2 cells

Iron overload often occurs during blood transfusion and iron supplementation, resulting in the presence of non-transferrin-bound iron(NTBI) in host plasma and damage to multiple organs, but effects on the intestine have rarely been reported. In this study, an iron overload mouse model with plasma NTBI was established by intraperitoneal injection of iron dextran. We found that plasma NTBI damaged intestinal morphology, caused intestinal oxidative stress injury and reactive oxygen species(ROS) accumulation,and induced intestinal epithelial cell apoptosis. In addition, plasma NTBI increased the relative abundance of Ileibacterium and Desulfovibrio in the cecum, while the relative abundance of Faecalibaculum and Romboutsia was reduced. Ileibacterium may be a potential microbial biomarker of plasma NTBI. Based on the function prediction analysis, plasma NTBI led to the weakening of intestinal microbiota function, significantly reducing the function of the extracellular structure. Further investigation into the mechanism of injury showed that iron absorption in the small intestine significantly increased in the iron group. Caco-2 cell monolayers were used as a model of the intestinal epithelium to study the mechanism of iron transport. By adding ferric ammonium citrate(FAC, plasma NTBI in physiological form) to the basolateral side, the apparent permeability coefficient(Papp) values from the basolateral to the apical side were greater than 3×10^(-6)cm s^(-1). Intracellular ferritin level and apical iron concentration significantly increased, and SLC39A8(ZIP8) and SLC39A14(ZIP14) were highly expressed in the FAC group.Short hairpin RNA(sh RNA) was used to knock down ZIP8 and ZIP14 in Caco-2 cells. Transfection with ZIP14-specific sh RNA decreased intracellular ferritin level and inhibited iron uptake. These results revealed that plasma NTBI may cause intestinal injury and intestinal flora dysbiosis due to the uptake of plasma NTBI from the basolateral side into the small intestine, which is probably mediated by ZIP14.

Studies on the fat mass and obesity-associated(FTO)gene and its impact on obesity-associated diseases

Obesity has become a major health crisis in the past∼50 years.The fat mass and obesity-associated(FTO)gene,identified by genome-wide association studies(GWAS),was first reported to be positively associated with obesity in humans.Mice with more copies of the FTO gene were observed to be obese,while loss of the gene in mice was found to protect from obesity.Later,FTO was found to encode an m6 A RNA demethylase and has a profound effect on many biological and metabolic processes.In this review,we first summarize recent studies that demonstrate the critical roles and regulatory mechanisms of FTO in obesity and metabolic disease.Second,we discuss the ongoing debates concerning the association between FTO polymorphisms and obesity.Third,since several small molecule drugs and micronutrients have been found to regulate metabolic homeostasis through controlling the expression or activity of FTO,we highlight the broad potential of targeting FTO for obesity treatment.Improving our understanding of FTO and the underlying mechanisms may provide new approaches for treating obesity and metabolic diseases.

Engineered Bacillus subtilis alleviates intestinal oxidative injury through Nrf2-Keap1 pathway in enterotoxigenic Escherichia coli(ETEC) K88-infected piglet

Engineered probiotics can serve as therapeutics based on their ability of produce recombinant immune-stimulating properties.In this study,we built the recombinant Bacillus subtilis WB800 expressing antimicrobial peptide KR32(WB800-KR32)using genetic engineering methods and investigated its protective effects of nuclear factor-E2-related factor 2(Nrf2)-Kelch-like ECH-associated protein 1(Keap1)pathway activation in intestinal oxidative disturbance induced by enterotoxigenic Escherichia coli(ETEC)K88 in weaned piglets.Twenty-eight weaned piglets were randomly distributed into four treatment groups with seven replicates fed with a basal diet.The feed of the control group(CON)was infused with normal sterilized saline;meanwhile,the ETEC,ETEC+WB800,and ETEC+WB800-KR32 groups were orally administered normal sterilized saline,5×10^(10)CFU(CFU:colony forming units)WB800,and 5×10^(10)CFU WB800-KR32,respectively,on Days 1-14 and all infused with ETEC K881×10^(10)CFU on Days 15-17.The results showed that pretreatment with WB800-KR32 attenuated ETEC-induced intestinal disturbance,improved the mucosal activity of antioxidant enzyme(catalase(CAT),superoxide dismutase(SOD),and glutathione peroxidase(GPx))and decreased the content of malondialdehyde(MDA).More importantly,WB800-KR32 downregulated genes involved in antioxidant defense(GPx and SOD1).Interestingly,WB800-KR32 upregulated the protein expression of Nrf2 and downregulated the protein expression of Keap1 in the ileum.WB800-KR32 markedly changed the richness estimators(Ace and Chao)of gut microbiota and increased the abundance of Eubacterium_rectale_ATCC_33656 in the feces.The results suggested that WB800-KR32 may alleviate ETEC-induced intestinal oxidative injury through the Nrf2-Keap1 pathway,providing a new perspective for WB800-KR32 as potential therapeutics to regulate intestinal oxidative disturbance in ETEC K88 infection.

甘草酸对体外鸡巨噬细胞免疫和杀菌功能的影响（英文）

目的:探究甘草酸能否激活体外鸡巨噬细胞并增强其免疫和吞噬杀菌功能。创新点:甘草酸通过核因子κB(NF-κB)和c-Jun氨基端激酶(JNK)信号通路提高一氧化氮(NO)和过氧化氢(H2O2)产生量,增强了其吞噬和杀菌的功能。方法:以不同浓度的甘草酸(0、12.5、25、50、100、200、400和800μg/ml)处理鸡巨噬细胞系HD11,采用荧光定量聚合酶链式反应(qP CR)和一氧化氮及过氧化氢测定试剂盒评价甘草酸对鸡巨噬细胞活化和免疫的影响,采用流式细胞技术和涂板计数法测定鸡巨噬细胞吞噬和杀菌能力。结论:甘草酸通过NF-κB和JNK信号通路激活鸡巨噬细胞,提高免疫细胞因子等基因的表达水平和NO及H_2O_2的产生量,从而增强了鸡巨噬细胞吞噬和清除胞内沙门氏菌的能力。

口服鼠李糖乳杆菌GG影响哺乳仔猪肠道屏障功能的研究（英文）

目的:探究新生仔猪口服鼠李糖乳杆菌GG对肠道屏障功能的影响。创新点:新生仔猪早期口服鼠李糖乳杆菌GG可明显改善其断奶前肠道菌群结构及免疫屏障功能。方法:二十四头新生仔猪分为对照组和实验组:对照组仔猪在出生后第1、3、5天口服2 mL 0.1 g/mL的脱脂牛奶;而实验组仔猪口服等体积的含有活鼠李糖乳杆菌GG的脱脂牛奶。饲喂25天后,收集仔猪血清、肠道粘膜和盲肠内容物等样品。通过分析肠道菌群、紧密连接蛋白和细胞因子等指标,评价鼠李糖乳杆菌对肠道屏障功能的影响。结论:在正常生理条件下,新生仔猪口服鼠李糖乳杆菌GG可明显改变肠道菌群结构。此外,鼠李糖乳杆菌GG还可增加仔猪肠道的通透性,并通过调控抗菌肽、细胞因子和趋化因子的分泌以改善肠道的免疫屏障功能。