Multi-omics analysis reveals the molecular regulatory network underlying the prevention of Lactiplantibacillus plantarum against LPS-induced salpingitis in laying hens

Background Salpingitis is one of the common diseases in laying hen production, which greatly decreases the economic outcome of laying hen farming. Lactiplantibacillus plantarum was effective in preventing local or systemic inflammation, however rare studies were reported on its prevention against salpingitis. This study aimed to investigate the preventive molecular regulatory network of microencapsulated Lactiplantibacillus plantarum(MLP) against salpingitis through multi-omics analysis, including microbiome, transcriptome and metabolome analyses.Results The results revealed that supplementation of MLP in diet significantly alleviated the inflammation and atrophy of uterus caused by lipopolysaccharide(LPS) in hens(P < 0.05). The concentrations of plasma IL-2 and IL-10 in hens of MLP-LPS group were higher than those in hens of LPS-stimulation group(CN-LPS group)(P < 0.05). The expression levels of TLR2, MYD88, NF-κB, COX2, and TNF-α were significantly decreased in the hens fed diet supplemented with MLP and suffered with LPS stimulation(MLP-LPS group) compared with those in the hens of CN-LPS group(P < 0.05). Differentially expressed genes(DEGs) induced by MLP were involved in inflammation, reproduction, and calcium ion transport. At the genus level, the MLP supplementation significantly increased the abundance of Phascolarctobacterium, whereas decreased the abundance of Candidatus_Saccharimonas in LPS challenged hens(P < 0.05). The metabolites altered by dietary supplementation with MLP were mainly involved in galactose, uronic acid, histidine, pyruvate and primary bile acid metabolism. Dietary supplementation with MLP inversely regulates LPSinduced differential metabolites such as Lyso PA(24:0/0:0)(P < 0.05).Conclusions In summary, dietary supplementation with microencapsulated Lactiplantibacillus plantarum prevented salpingitis by modulating the abundances of Candidatus_Saccharimonas, Phascolarctobacterium, Ruminococcus_torques_group and Eubacterium_hallii_group while downregulating the levels of plasma metabolites, p-tolyl sulfate, o-cresol and N-acetylhistamine and upregulating S-lactoylglutathione, simultaneously increasing the expressions of CPNE4, CNTN3 and ACAN genes in the uterus, and ultimately inhibiting oviducal inflammation.

Rosemary extract improves egg quality by altering gut barrier function,intestinal microbiota and oviductal gene expressions in late-phase laying hens

Background Rosemary extract(RE)has been reported to exert antioxidant property.However,the application of RE in late-phase laying hens on egg quality,intestinal barrier and microbiota,and oviductal function has not been systematically studied.This study was investigated to detect the potential effects of RE on performance,egg quality,serum parameters,intestinal heath,cecal microbiota and metabolism,and oviductal gene expressions in late-phase laying hens.A total of 21065-week-old“Jing Tint 6”laying hens were randomly allocated into five treatments with six replicates and seven birds per replicate and fed basal diet(CON)or basal diet supplemented with chlortetracycline at 50 mg/kg(CTC)or RE at 50 mg/kg(RE50),100 mg/kg(RE100),and 200 mg/kg(RE200).Results Our results showed that RE200 improved(P<0.05)Haugh unit and n-6/n-3 of egg yolk,serum superoxide dismutase(SOD)compared with CON.No significant differences were observed for Haugh unit and n-6/n-3 of egg yolk among CTC,RE50,RE100 and RE200 groups.Compared with CTC and RE50 groups,RE200 increased serum SOD activity on d 28 and 56.Compared with CON,RE supplementation decreased(P<0.05)total cholesterol(TC)level.CTC,RE100 and RE200 decreased(P<0.05)serum interleukin-6(IL-6)content compared with CON.CTC and RE200 increased jejunal m RNA expression of ZO-1 and Occludin compared with CON.The biomarkers of cecal microbiota and metabolite induced by RE 200,including Firmicutes,Eisenbergiella,Paraprevotella,Papillibacter,and butyrate,were closely associated with Haugh unit,n-6/n-3,SOD,IL-6,and TC.PICRUSt2 analysis indicated that RE altered carbohydrate and amino acid metabolism of cecal microbiota and increased butyrate synthesizing enzymes,including 3-oxoacid Co A-transferase and butyrate-acetoacetate Co A-transferase.Moreover,transcriptomic analysis revealed that RE200 improved gene expressions and functional pathways related to immunity and albumen formation in the oviductal magnum.Conclusions Dietary supplementation with 200 mg/kg RE could increase egg quality of late-phase laying hens via modulating intestinal barrier,cecal microbiota and metabolism,and oviductal function.Overall,RE could be used as a promising feed additive to improve egg quality of laying hens at late stage of production.

Prevotella and succinate treatments altered gut microbiota,increased laying performance,and suppressed hepatic lipid accumulation in laying hens

Background This work aimed to investigate the potential benefits of administering Prevotella and its primary metabolite succinate on performance,hepatic lipid accumulation and gut microbiota in laying hens.Results One hundred and fifty 58-week-old Hyline Brown laying hens,with laying rate below 80%and plasma triglyceride(TG)exceeding 5 mmol/L,were used in this study.The hens were randomly allocated into 5 groups and subjected to one of the following treatments:fed with a basal diet(negative control,NC),oral gavage of 3 mL/hen saline every other day(positive control,PC),gavage of 3 mL/hen Prevotella melaninogenica(10^(7)CFU/mL,PM)or 3 mL/hen Prevotella copri(10^(7)CFU/mL,P.copri)every other day,and basal diet supplemented with 0.25%sodium succinate(Succinate).The results showed that PM and P.copri treatments significantly improved laying rate compared to the PC(P<0.05).The amount of lipid droplet was notably decreased by PM,P.copri,and Succinate treatments at week 4 and decreased by P.copri at week 8(P<0.05).Correspondingly,the plasma TG level in Succinate group was lower than that of PC(P<0.05).Hepatic TG content,however,was not significantly influenced at week 4 and 8(P>0.05).PM treatment increased(P<0.05)the mRNA levels of genes PGC-1βand APB-5B at week 4,and ACC and CPT-1 at week 8.The results indicated enhanced antioxidant activities at week 8,as evidenced by reduced hepatic malondialdehyde(MDA)level and improved antioxidant enzymes activities in PM and Succinate groups(P<0.05).Supplementing with Prevotella or succinate can alter the cecal microbiota.Specifically,the abundance of Prevotella in the Succinate group was significantly higher than that in the other 4 groups at the family and genus levels(P<0.05).Conclusions Oral intake of Prevotella and dietary supplementation of succinate can ameliorate lipid metabolism of laying hens.The beneficial effect of Prevotella is consistent across different species.The finding highlights that succinate,the primary metabolite of Prevotella,represents a more feasible feed additive for alleviating fatty liver in laying hens.

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.

Chronic heat stress induces renal fibrosis and mitochondrial dysfunction in laying hens

Background Heat stress in laying hens negatively affects egg production and shell quality by disrupting the homeo-stasis of plasma calcium and phosphorus levels.Although the kidney plays an important role in calcium and phos-phorus homeostasis,evidence regarding the effect of heat stress on renal injury in laying hens is yet to be elucidated.Therefore,the aim of this study was to evaluate the effects of chronic heat stress on renal damage in hens during laying periods.Methods A total of 16 white-leghorn laying hens(32 weeks old)were randomly assigned to two groups(n=8).One group was exposed to chronic heat stress(33°C for 4 weeks),whereas the other group was maintained at 24°C.Results Chronic heat exposure significantly increased plasma creatinine and decreased plasma albumin levels(P<0.05).Heat exposure also increased renal fibrosis and the transcription levels of fibrosis-related genes(COLA1A1,αSMA,and TGF-β)in the kidney.These results suggest that renal failure and fibrosis were induced by chronic heat exposure in laying hens.In addition,chronic heat exposure decreased ATP levels and mitochondrial DNA copy number(mtDNA-CN)in renal tissue,suggesting that renal mitochondrial dysfunction occurs under conditions of heat stress.Damaged mitochondria leak mtDNAs into the cytosol and mtDNA leakage may activate the cyclic GMP-AMP synthase(cGAS)stimulator of interferon genes(STING)signaling pathway.Our results showed that chronic heat exposure activated the cGAS-STING pathway as indicated by increased expression of MDA5,STING,IRF7,MAVS,and NF-κB levels.Furthermore,the expression of pro-inflammatory cytokines(IL-12)and chemokines(CCL4 and CCL20)was upregulated in heat-stressed hens.Conclusions These results suggest that chronic heat exposure induces renal fibrosis and mitochondrial damage in laying hens.Mitochondrial damage by heat stress may activate the mtDNA-cGAS-STING signaling and cause subse-quent inflammation,which contributes to the progression of renal fibrosis and dysfunction.

Adjusting phosphate feeding regimen according to daily rhythm increases eggshell quality via enhancing medullary bone remodeling in laying hens

Background Body phosphorus metabolism exhibits a circadian rhythm over the 24-h daily cycle.The egg laying behavior makes laying hens a very special model for investigating phosphorus circadian rhythms.There is lack of information about the impact of adjusting phosphate feeding regimen according to daily rhythm on the phosphorus homeostasis and bone remodeling of laying hens.Methods and results Two experiments were conducted.In Exp.1,Hy-Line Brown laying hens(n=45)were sampled according the oviposition cycle(at 0,6,12,and 18 h post-oviposition,and at the next oviposition,respectively;n=9 at each time point).Diurnal rhythms of body calcium/phosphorus ingestions and excretions,serum calcium/phosphorus levels,oviduct uterus calcium transporter expressions,and medullary bone(MB)remodeling were illustrated.In Exp.2,two diets with different phosphorus levels(0.32%and 0.14%non-phytate phosphorus(NPP),respectively)were alternately presented to the laying hens.Briefly,four phosphorus feeding regimens in total(each included 6 replicates of 5 hens):(1)fed 0.32%NPP at both 09:00 and 17:00;(2)fed 0.32%NPP at 09:00 and 0.14%NPP at 17:00;(3)fed 0.14%NPP at 09:00 and 0.32%NPP at 17:00;(4)fed 0.14%NPP at both 09:00 and 17:00.As a result,the regimen fed 0.14%NPP at 09:00 and 0.32%NPP at 17:00,which was designed to strengthen intrinsic phosphate circadian rhythms according to the findings in Exp.1,enhanced(P<0.05)MB remodeling(indicated by histological images,serum markers and bone mineralization gene expressions),elevated(P<0.05)oviduct uterus calcium transportation(indicated by transient receptor potential vanilloid 6 protein expression),and subsequently increased(P<0.05)eggshell thickness,eggshell strength,egg specific gravity and eggshell index in laying hens.Conclusions These results underscore the importance of manipulating the sequence of daily phosphorus ingestion,instead of simply controlling dietary phosphate concentrations,in modifying the bone remodeling process.Body phosphorus rhythms will need to be maintained during the daily eggshell calcification cycle.

Effects of Desmodium uncinatum Leaf Meal in the Diet on Lohmann Brown Hens’ Laying Performance and Eggs Quality

A cheap, safe and available alternative protein source to soybean is a huge priority for the developing world. This study is aimed at evaluating the effects of Desmodium uncinatum leaf meal incorporation levels in the diet on Lohmann Brown hens’ laying performances. For this purpose, 60 Lohmann Brown hens aged 33 weeks with an average weight of 1.78 kg were randomly assigned to 3 treatment groups of 20 birds each with 4 replicates. The experimental treatments consisted of DULM 0, DULM5, and DULM10 containing 0, 5 and 10 kg of D. uncinatum leaf meal (DULM) per 100 kg of feed respectively. The results reveal that except for the weights of hens, laying performances were significantly affected (p < 0.05) by DULM in the diet. Daily feed intake was lower with 10% incorporation of DULM in the diet as compared to a diet with 5% DULM which recorded the highest value for this parameter. Moreover, the highest value for the egg weight, egg mass, yolk index, albumen index and egg production were recorded with diet DULM5. On the other hand, the diet with 10% of DULM (DULM10) recorded the highest relative shell weight, feed conversion ratio and egg price. Nevertheless, the mortality rate found in this study was 0% for all treatments confirming the good quality of DULM as protein feedstuff for layer. It was concluded that incorporation levels of D. uncinatum leaves at 5% in the diet improved the laying performances of Lohmann Brown hens.

Performance Evaluation of Manually Operated Mulch Laying Machine on Different Soil Conditions

A machine with manual operation for mulch-laying machine with a punching arrangement was developed,and its performance was assessed at three different mulch paper thicknesses(15 m,20 m and 25 m),three different disc angles(35 degrees,40 degrees,and 45 degrees),three different punch spacings(250 mm,500 mm,and 1000 mm),and three different forward speeds(1.3 km/h,1.5 km/h,1.7 km/h)to investigate their effects on field capacity,effective field efficiency,and punching efficiency.Utilizing randomised block design and response surface methods,the experimental plan for optimization was created.All of the independent variables’combined effects on the dependent variables were found to be statistically significant.The influence of operating speed and mulch paper thickness was found to be the most significant on the dependent variable.The effective field capacity and field efficiency increased from 0.11 ha/h to 0.19 ha/h and 72.04 percent to 89.51 percent,respectively,by increasing mulch paper thickness from 15μm to 25μm and operating speed from 1.3 km/h to 1.7 km/h,whereas punching efficiency fell from 85.18 percent to 84.40 percent.Mulch paper of 15μm and a disc angle value of 40 degrees were optimised from the independent factors that were chosen for optimal soil covering over the laid plastic mulch sheet.Punching efficiency was maximised with performance optimised at 500 mm punch spacing.Additionally,the machine operated more efficiently at 1.5 km/h.

Soya saponin improves egg-laying performance and immune function of laying hens

Background:Soya saponin(SS),an active compound in soybean meals,has been widely studied in the medical field.However,it was considered as an anti-nutritional factor in poultry diets.The objective of this experiment was to measure the effects of dietary SS using three dietary treatments on egg-laying performance and immune function of laying hens.Birds were fed a low soybean meal basal diet(CON),a low-SS diet(50 SS)containing 50mg/kg SS,or a high-SS diet(500 SS)containing 500 mg/kg SS for 10 weeks.At the end of the 5th and 10th week of the trial,samples were collected for analysis.Results:Results showed that with 50 mg/kg SS supplementation,the egg production rate,feed conversion ratio(FCR),and eggshell quality tended to be improved.Serum follicle stimulating hormone(FSH)and Interleukin-4(IL-4)levels were also elevated as well as the peripheral blood LPS stimulation index,the proportion of B lymphocytes,and antibody titer of bovine serum albumin(BSA).We also found that mRNA levels of follicle stimulating hormone receptor(FSHR)in ovarian,nuclear transcription factor kappa B(NF-κB),Transforming growth factor(TGF-β)and interferonγ(IFN-γ)in spleen were up-regulated at the end of the trial.Additionally,dietary 50 mg/kg SS improved the ileal flora via up-regulating the relative abundance of Lactobacillus,Romboutsia and Lactobacillus delbrueckii.Although the immune related indicators were improved with 500 mg/kg SS supplemented,it seemed to have a negative influence on the laying-performance.Specifically,serum alanine aminotransferase(ALT),alkaline phosphatase(ALP),and the ratio of IFN-γto IL-4 were increased in the 500 SS group at the end of the trial.The m RNA levels of gonadotropin releasing hormone 1(GnRH1)in Hypothalamus,the estrogen related receptor(ERR)in ovaries were downregulated as well as the egg production rate during the trial with 500 mg/kg SS supplemented.Conclusions:The egg production performance was improved by dietary supplemented with 50 mg/kg SS via increasing ovarian FSHR transcription level and serum estrogen level.A beneficial shift in intestinal microflora was recorded,and the immune function of laying hens was also improved with 50 mg/kg SS supplementation.Surprisingly,the long-term supplementation of 500 mg/kg SS exerted a negative impact on the laying performance and physiological functions of the liver of laying hens.

Mechanism of the Effect of Bacillus coagulans on Production Performance of Laying Hens in Late Laying Period

[Objective]The paper was to investigate the mechanism of Bacillus coagulans preparation in improving production performance of laying hens in late period of laying.[Method]A total of 648 individuals of"Jingfen 1"laying hens(66 weeks of age)in late period of laying were randomly divided into four groups,six replicates each group,and each group had 27 individuals of laying hens.The laying hens in control group were fed with basal diet,and those in experimental groupsⅠ,Ⅱ and Ⅲ were fed with the basal diets added with 3.33×10^6,1×10^7,3.33×10^7 CFU/g B.coagulans,respectively.The pretrial lasted one week,and the formal test lasted eight weeks.[Result]Compared with the control group,the laying rate in group Ⅰ increased significantly,and the feed-gain ratio in experimental group Ⅱ decreased noticeably while the spleen index increased remarkably;the alkaline phosphatase activities and blood calcium content in experimental groups Ⅰ,Ⅱ and Ⅲ increased significantly(P<0.05),while the triglyceride content decreased remarkably(P<0.05);the urea content in experimental groups Ⅰ and Ⅱ decreased obviously(P<0.05).Adding B.coagulans significantly increased the specific activity of amylase,lipase and protease in various intestinal mucosa of small intestine(P<0.05);adding B.coagulans significantly increased the content of follicle-stimulating hormone(FSH)and estradiol(E2),and significantly increased the mRNA expression of follicle stimulating hormone receptor gene(FSHR).[Conclusion]B.coagulans preparation could significantly improve the production performance of laying hens in late period of laying;appropriately enhance the immune capacity of laying hens;improve the serum biochemical indicators;increase the activity of digestive enzymes in the small intestine;and promote the release of gonadal hormone and the mRNA expression of FSHR gene in the ovary.

Effects of Quercetin on Performance and Lipid Metabolism in Laying Hens

Quercetin has a variety of biological activities. However, its application as a feed additive in poultry production is limited. In this study, the effects of quercetin on performance and lipid metabolism in Hessian laying hens were investigated and the possibility of quercetin to decrease cholesterol in eggs and Low Density Lipoprotein cholesterol（LDL-cholesterol） in laying hens were explored. A total of 240 laying hens at 29-week-old were randomly assigned to four treatments, with six replicates of 10 birds per replicate. The treatments were fed with basal diet supplemented with 0, 0.2, 0.4 and 0.6 g · kg-1 quercetin for 8 weeks. Layer performance, lipid concentration in livers, yolks and serum were measured at the end of the experiment. The results showed that laying rate had a quadratic correlation with the level of quercetin（p=0.056） and was maximized by the supplement level of 0.2 g · kg-1 diet. When the level of dietary quercetin increased, feed-egg ratio decreased. Crude fat and the total cholesterol of livers decreased by supplementation of quercetin. The total cholesterols of yolks also decreased and had a quadratic correlation with the levels of quercetin（p=0.019）. Insulin increased by supplement of quercetin. In conclusion, quercetin improved the performance and reduced egg yolk cholesterol by affecting serum hormone levels.

Effects of Yeast Chromium Combined with Probiotics on Laying Performance,Intestinal Microflora and Digestive Enzyme Activities of Laying Hens in Summer

[ Objective] To study the effects of yeast chromium combined with probiotics on laying performance, intestinal microflora and digestive enzyme activities of laying hens. [ Method] A total of 432 healthy 40-week-old Roman brown laying hens were randomly divided into nine groups. Laying hens in control group were fed with basal diet, laying hens in test groups were fed with basal diet combining with different levels of yeast chromium （0,400,600 μg/kg） or probiotics （0,400,800 mg/kg）. [ Result] Yeast chromium, probiotics or yeast chromium combined with probiotics adding into feeds could significantly increased the laying rate （ P 〈 0.05 ） and significantly reduced feed-egg rate （ P 〈 0.05 ）. Joint use of yeast chromium and probiotics had significant interaction effects on laying rate and feed conversion rate（P 〈 0. 05）. Adding of yeast chromium combined with probiotics in diets could significantly increased the amount of Lactobacillus and Bifidobacterium （P 〈0.05 ） in ileum and cecum of laying hens, and significantly reduced the amount of Escherichia coli and Salmonella （P 〈0.05 ） , in addition, the activity of total protease, lipase and amylase in duodenal contents were significantly increased （ P 〈 0.05 ）. Joint use of yeast chromium and probioties had sig- nificant interaction effects on the activity of total protease, lipase and amylase in laying hens duodenal contents （ P 〈 0.05 ）. [ Conclusion ] Joint adding of 600μg/kg yeast chromium and 800 mg/kg probiotics in diets in summer had best effects on laying performance, intestinal microflora and digestive enzyme activities of laying hens.

Force Analysis and Curve Design for Laying Pipe in Loop Laying Head of Wire Rod Mills

Laying head is a high-precision engineering device in hot-rolled high speed wire rod production line. Previously research works are focused on the laying pipe wear-resisting. Laying pipe curve design method based on wire rod kinematics and dynamics analyses are not reported before. In order to design and manufacture the laying pipe, the motion and force process of the wire rod in the laying pipe should be studied. In this paper, a novel approach is proposed to investigate the force modeling for hot-rolled wire rod in laying pipe. An idea of limited element method is used to analysis and calculates the forces between laying pipe inner surface and wire rod. The design requirements of laying pipe curve for manufacturing are discussed. The kinematics and dynamics modeling for numerical calculation are built. A laying pipe curve equation is proposed by discussing design boundary conditions. Numerical results with di erent laying pipe curves design parameters are plotted and compared. The proposed approach performs good result which can be applied for laying pipe curve design and analysis for engineering application.

Laying a Solid Foundation,Expanding Development Space——Foundation Stone Laying Ceremony of ZTE R&D Building Held in Shenzhen

Earth was broken for the ZTE R&D Building at theheadquarters on October 28, 2003. Hou Weigui,President of ZTE, delivered a speech on the cere-mony. He said:“As part of the development strate-gy of ZTE, the new building will play an importantrole in improving the overall R&D arrangement andthe environment of production and R&D, reducingthe operating cost and enhancing the image of thewhole company.”The R&D building will be the Shenzhen R&D

An Overview of Deepwater Pipeline Laying Technology

The technology and methods involved in pipeline laying in shallow water have evolved to the level of routine and commonplace. However, regarding the unexpected deepwater complexity, the traditional pipeline laying techniques have to confront many new challenges arisen from the increase of the water depth, diameter of the pipe and the welding difficuhy, all of which should be modified and/or innovated based on the existed mature experiences. The purpose of this investigation is to outline the existing and new engineering laying techniques and the associated facilities, which can provide some significant information to the related research. In the context, the latest deepwater pipeline laying technology and pipe laying barges of the renowned companies from Switzerland, Norway, Italy etc., are introduced and the corresponding comparison and discussion are presented as well.

Dietary oregano essential oil supplementation improves intestinal functions and alters gut microbiota in latephase laying hens

Background:Dietary essential oil(EO)supplementation can exert favorable effects on gut health in broilers.However,it is unknown whether EO could improve intestinal functions,consequently beneficial for egg performance and quality in late-phase laying hens.This study was aimed to investigate the potential effects of EO on production performance,egg quality,intestinal health and ileal microbiota of hens in the late phase of production.A total of 28860-week-old Hy-line Brown laying hens were randomly divided into 4 groups and fed a basal diet(control)or basal diets supplemented with oregano EO at 100,200 and 400 mg/kg(EO100,EO200 and EO400).Results:Dietary EO supplementation resulted in a quadratic decrease(P<0.05)in feed conversion ratio with lower(P<0.05)feed conversion ratio in EO200 group than the control during weeks 9–12 and 1–12 of the trial.Compared to the control,EO addition resulted in higher(P<0.05)eggshell thickness at the end of week.4,8 and 12 and higher(P<0.05)chymotrypsin activity.There was a quadratic elevation(P<0.05)in ileal chymotrypsin and lipase activity,along with a linear increase in villus height to crypt depth ratio.Quadratic declines(P<0.05)in mRNA expression of IL-1β,TNF-α,IFN-γ and TLR-4,concurrent with a linear and quadratic increase(P<0.05)in ZO-1 expression were identified in the ileum with EO addition.These favorable effects were maximized at medium dosage(200 mg/kg)of EO addition and intestinal microbial composition in the control and EO200 groups were assessed.Dietary EO addition increased(P<0.05)the abundances of Burkholderiales,Actinobacteria,Bifidobacteriales,Enterococcaceae and Bacillaceae,whereas decreased Shigella abundance in the ileum.Conclusions:Dietary EO addition could enhance digestive enzyme activity,improve gut morphology,epithelial barrier functions and modulate mucosal immune status by altering microbial composition,thus favoring feed efficiency and eggshell quality of late-phase laying hens.

Ascorbic acid synthesis and transportation capacity in old laying hens and the effects of dietary supplementation with ascorbic acid

Background: Laying hens over 75 weeks of age commonly show great declines in immunity and production performance.It is unclear whether these declines can be relieved by supplementing with ascorbic acid(AA) in feed.Two trials were conducted to investigate the synthesis and metabolism of AA in layers of different ages and the effects of dietary supplemental AA on the performance and the immune and antioxidant statuses of 78 weeks old hens.Methods: In Exp.1,equal numbers(24 hens) of 35 weeks old(Young) and 75 weeks old(Old) layers were fed the same diet without AA supplementation for 4 weeks.In Exp.2,360 healthy 78 weeks old laying hens were randomly assigned to 4 treatments(basal diet supplemented with 0,0.25,0.5,or 1 g AA/kg diet) in an 8-week feeding trial.Results: The old hens tended to have decreased L-gulonolactone oxidase(GLO) synthase activity in the kidney and liver than that of the young hens(P = 0.07 and P = 0.05,respectively).Compared with the young hens,the old hens had lower hepatic antioxidant capacity allowing for the lower thioredoxin(TXN),thioredoxin reductase(TXNR) and cytochrome b5 reductase(CYB5 R) gene expression(P < 0.05),whereas increased sodium-dependent vitamin C transporter(SVCT) 1 expression levels in the ileum and kidney and enhanced splenic and hepatic AA concentrations(P < 0.05).Dietary supplementation with AA significantly decreased GLO enzyme activity but increased splenic AA concentration and anti-bovine serum albumin IgG levels(P < 0.05) and tended to increase CD4+T lymphocyte numbers(P = 0.06) in serum.Supplementation of 0.25 g AA/kg diet significantly increased hepatic total antioxidant capacity(T-AOC,P < 0.05) relative to the control group.Conclusions: Laying hens could synthesize AA in both the kidney and the liver,though the GLO enzyme activities were 100 times greater in kidneys than in livers.The old laying hens had greater absorption and reabsorption capacity and higher AA retention in some tissues that did the young hens.Dietary supplementation of AA can improve the health of old layers by enhancing immunity and antioxidant capacity.

Dietary betaine activates hepatic VTGⅡexpression in laying hens associated with hypomethylation of GR gene promoter and enhanced GR expression

Background: Vitellogenin(VTG) is a precursor of egg yolk proteins synthesized within the liver of oviparous vertebrates. Betaine is an important methyl donor that is reported to improve egg production of laying hens with an unclear mechanism. In the present study, we fed betaine-supplemented diet(0.5%) to laying hens for 4 wk and investigated its effect on VTGII expression in the liver.Results: Betaine did not affect chicken weight, but significantly(P < 0.05) increased egg laying rate accompanied with a significant(P < 0.05) increase in hepatic concentration and plasma level of VTGI. Plasma estrogen level did not change,but the hepatic expression of estrogen receptor α(ERα) mR NA was significantly(P < 0.05) up-regulated. Betaine did not affect the protein content of ERα, but significantly(P < 0.05) increased hepatic expression of glucocorticoid receptor(GR)at both mR NA and protein levels. Also, ERα/GR interaction tended to be enhanced in the liver nuclear lysates of betainesupplemented hens as determined by co-immunoprecipitation. Furthermore, dietary betaine supplementation significantly increased(P < 0.05) the hepatic expression of methyl-transfer enzymes, such as BHMT, GNMT, and DNMT1,which was associated with higher SAM/SAH ratio and hypomethylation of GR promoter regions.Conclusions: Betaine activates hepatic VTGII expression in association with modified DNA methylation of GR gene promoter, GR expression and ERα/GR interaction. Activation of hepatic VTGII expression may contribute, at least partly, to improved egg production in betaine-supplemented hens.

Effect of dietary nonphytate phosphorus on laying performance and small intestinal epithelial phosphate transporter expression in Dwarf pink-shell laying hens

This study examined the effects of various levels of dietary nonphytate phosphorus on laying performance and the expression patterns of phosphorus metabolism related genes in Dwarf pink-shell Jaying hens. A total of 405 28-week-old Dwarf pink-shell laying hens were fed the same corn-soybean basal meals but containing 0.20%, 0.25%, 0.30%, 0.35% or 0.40% nonphytate phosphorus. The results showed that feed intake, egg production, and average egg weights were quadratically correlated with dietary nonphytate phosphorus content （P 〈 0.05）, and the highest egg production, feed intake and average egg weights were achieved when dietary nonphytate phosphorus was at 0.3% （P 〈 0.05）. mRNA expression of intestinal sodium phosphorus co-transporter linearly decreased when dietary nonphytate phosphorus increased, mRNA and protein expression of intestinal calbindin and vitamin D receptor correlated quadratically with dietary nonphytate phosphorus, and the highest expression was found when dietary available phosphorus was at 0.2,5% to 0.3%. In conclusion, the ideal phosphorus requirement for Dwarf pink-shell layer hens is estimated to be 0.3% in a corn-soybean diet. With this level of phosphorus supplementation, calbindin and vitamin D receptor reached their highest expression.

Effects of Fermentation Product Containing Phytase on Productive Performance,Egg Quality,and Phosphorous Apparent Metabolism of Laying Hens Fed Different Levels of Phosphorus

This study investigated the effects of fermentation product containing phytase （FPP） that was fermented using waste vinegar residue （WVR） as substrate from Aspergillusficuum NTG-23 on productive performance, egg quality, and phosphorus apparent metabolism of laying hens. First, 375 22-wk-old Jinghong hens were allocated into 5 treatments （5 replicates of 15 hens each） in an 8-wk experiment for evaluating the parameters of productive performance, egg quality, serum, and tibia. Experimental diets contained 4% FPP and 96% corn-soybean diet. The levels of dicalciurn phosphate （DCP） were 1.34, 1.01, 0.67, 0.34 and 0%. Next, thirty 31-wk-old Jinghong hens were fed 5 types of diets for evaluating phosphorous apparent metabolism rate. Egg productive rate, egg weight, feed conversion ratio, Haugh unit, egg albumen height, serum calcium, tibia ash, tibia ash calcium and tibia breaking strength were not different significantly among 5 treatments. The significant difference of average daily feed intake was not appeared when the DCP content of corn-soybean-FPP diet was reduced to 0.67%; the eggshell hardness, eggshell thickness and serum phosphorus were not reduced significantly until the DCP content of corn-soybean-FPP diet was reduced to 0.34%. The yolk color was improved when the laying hens fed deficient DCP corn-soybean-FPP diet. A 22.14% reduction in excreta phosphorus was observed when the laying hens fed low phosphorus （0.67% DCP） corn-soybean-FPP diet. A 30% elevation of phosphorus apparent metabolism rate was obtained when the DCP content of corn-soybean-FPP diet was decreased from 1.34 to 1.01%. The reducing cost of layer diet was totalized about 120 CNY 1000 kg1 diet when the content of DCP was 0.67% in corn-soybean-FPP diet. These results indicated that FPP could be applied in laying hen as a potential, cost-effective and rational application of WVR.