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.

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.

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.

Effect of Supplemental Oregano Essential Oils in Diets on Production Performance and Relatively Intestinal Parameters of Laying Hens

This study was designed to investigate the effects of dietary oregano essential oils on 150 30-week-old Hy-Line Layers’ productive performance, egg quality characteristics, cecal microbiota, digestive enzyme activities, mucosa structure of the duodenum and jejunum and glucose and small peptides transporters expression in the duodenum and jejunum. All hens were allocated into one of five different groups: negative control (NC;basal diet only), antibiotics control (CS;basal diet plus 100 mg/kg of 10% colistin sulfate) and treatments I, II, and III (basal diet supplemented with 50, 100 and 150 mg/kg oregano essential oils, respectively). The results showed that the laying rate, average egg weight, feed conversion ratio and the activities of amylase and trypsin were significantly improved by a diet supplemented with 100 mg/kg oregano essential oils (P 0.05), significantly increased duodenum villus-height-to-crypt-depth ratios (P 0.05).

Evaluation of a laying-hen tracking algorithm based on a hybrid support vector machine

Background： Behavior is an important indicator reflecting the welfare of animals. Manual analysis of video is the most commonly used method to study animal behavior. However, this approach is tedious and depends on a subjective judgment of the analysts. There is an urgent need for automatic identification of individual animals and automatic tracking is a fundamental part of the solution to this problem.Results： In this study, an algorithm based on a Hybrid Support Vector Machine（HSVM） was developed for the automated tracking of individual laying hens in a layer group. More than 500 h of video was conducted with laying hens raised under a floor system by using an experimental platform. The experimental results demonstrated that the HSVM tracker outperformed the Frag（fragment-based tracking method）, the TLD（Tracking-Learning-Detection）,the PLS（object tracking via partial least squares analysis）, the Mean Shift Algorithm, and the Particle Filter Algorithm based on their overlap rate and the average overlap rate.Conclusions： The experimental results indicate that the HSVM tracker achieved better robustness and state-of-theart performance in its ability to track individual laying hens than the other algorithms tested. It has potential for use in monitoring animal behavior under practical rearing conditions.

Effects of dietary phosphorous supplementation on laying performance,egg quality,bone health and immune responses of laying hens challenged with Escherichia coli lipopolysaccharide

Background: Phosphorus is an essential nutrient to maintain poultry health and performance. The objective of this study was to evaluate the effect of dietary phosphorus levels on egg production, egg quality, bone health, immune responses of laying hens challenged with Escherichia coli lipopolysaccharide.Methods: Three hundred laying hens at 28 wk were randomly divided into 2 dietary treatments with 10 replicates of 15 birds. The wheat-soybean based diets contained either 0.12% or 0.4% non-phytate phosphorus(NPP). At 32 wk of age, all the birds of each dietary treatment were injected into the abdomen with 1.5 mg/kg body weight(BW) of either LPS or saline once a day at 24-h intervals for continuous 9 d. The performance of laying hens was evaluated for 9 d. The eggs after the fifth injection were collected to value the egg quality. Three hours after the first injection, blood was collected to measure serum metabolite and immune response associated parameters.Three hours after the fifth injection, the hens were euthanized to obtain tibia, cecal tonsils and jejunum.Results: Compared with saline-injected hens, LPS-injected hens had lower feed intake and egg production(P < 0.05).Eggshell thickness, strength, albumin height and Haugh unit were significantly increased in LPS-injected hens compared with saline-injected hens(P < 0.05). Furthermore, laying hens challenged with LPS had lower villious height/crypt depth ration than those received saline. Serum calcium, phosphorus and SOD activities significantly decreased in the LPS-injected hens compared with the control(P < 0.05). LPS up-regulated expression of IL-1β, IL-6 and IL-10 in cecum, and serum concentration of MDA, IL-1β and IL-6(P < 0.05), whereas 0.40% dietary non-phytate phosphorus supplementation significantly increased(P < 0.05) villi height/crypt depth ratio, decreased(P < 0.05) serum MDA and IFN-γ concentration compared with the 0.12% non-phytate phosphorus group.Conclusion: In summary, this study demonstrates that 0.40% dietary non-phytate phosphorus supplementation significantly increased calcium and phosphorus levels of eggshell, increased villi height/crypt depth ratio, decreased serum MDA and IFN-γ concentration compared with the 0.12% non-phytate phosphorus groups. The results indicate that high level of dietary non-phytate phosphorus exerts a potential effect in alleviating systemic inflammation of LPSchallenged laying hens.

Dietary L-citrulline supplementation modulates nitric oxide synthesis and antioxidant status of laying hens during summer season

Background:L-citrulline(L-Cit),a non-protein amino acid,has been implicated in several physiological functions including anti-inflammatory,anti-oxidative,and hypothermic roles,however,there is a paucity of information with regards to its potential in poultry production.Methods:This study was designed to investigate the effects of dietary L-Cit supplementation on the production performance,nitric oxide production,and antioxidant status of laying hens during summer period.Hy-Line Brown laying hens(n=288,34 weeks old)were allotted to four treatment,6 replicates of 12 chickens each.Dietary treatments of control(basal diets),0.25%,0.50%and 1.00%L-Cit supplementation were fed to chickens for eight(8)weeks.Production performance,free amino acid profiles,nitric oxide production,and antioxidant properties were measured.Blood samples were collected at the 4th and 8th weeks of the experiment.Results:Air temperature monitoring indicated an average daily minimum and maximum temperatures of 25.02°C and 31.01°C respectively.Dietary supplementation with L-Cit did not influence(P>0.05)the production performance,and rectal temperature of laying hens.Egg shape index was increased(P<0.05)with increasing levels of L-Cit.Serum-free content of arginine,citrulline,ornithine,tryptophan,histidine,GABA,and cystathionine were elevated,but taurine declined with L-Cit diets.Plasma nitric oxide(NOx)concentration was highest at 1%L-Cit.Likewise,nitric oxide synthase(NOS)activity for total NOS(tNOS)and inducible NOS(iNOS)were upregulated with increasing L-Cit levels,although,tNOS was not affected at the 4th week.Anti-oxidant enzymes including catalase and superoxide dismutase(SOD)were increased with L-Cit supplementation,however,SOD activity was unchanged at 4th week,while total anti-oxidant capacity increased at the 8th week.L-Cit supplementation attenuated the extent of lipid peroxidation,and also inhibited glutathione peroxidase activity.Conclusion:Dietary L-Cit supplementation modulated systemic arginine metabolism,nitric oxide synthesis,antioxidant defense system,and increased the egg shape index of laying hens during the summer season.1%L-Cit supplementation proved most effective in potentiating these effects and may be adopted for feed formulation strategies.

Effects of Macleaya cordata Extracts on Growth Performance and Immune Function of Immunosuppressed Laying Chicks

[Objective] The paper was to study the effects of Macleaya cordata extracts on growth performance and immune function of immunosuppressed laying chicks. [Method] A total of 120 1-day-old laying chicks with the body weight of (38.76±1.12) g were randomly divided into 3 groups with 40 replicates per treatment and 1 chick per replicate according to the principle of consistent body weight and half male and half female,namely control group, cyclophosphamide (CTX) group and M. cordata group. The trial lasted 21 d. The chicks in control group and CTX group were fed with basal diet, and those in M. cordata group were fed with the basal diet supplemented with 40 mg/kg M. cordata extract. On the 8^(th), 9^(th), 10^(th) and 17^(th) day of the trial, the chicks in CTX group and M. cordata group were intraperitoneally injected with 80 mg/kg BW CTX, and those in control group were intraperitoneally injected with equal dose of normal saline. [Result] (1) Compared with control group, the average daily gain (ADG) and average daily feed intake (ADFI) of chicks in CTX group reduced significantly;the feed gain ratio (F/G) increased significantly;the white blood cell count (WBC) decreased significantly;the alanine transaminase (ALT) and aspertate aminotransferase (AST) levels in the blood increased significant -ly;the total protein (TP) and globulin (GLOB) content in the blood decreased significantly;the serum malondialdehyde (MDA) content increased significantly;and the contents of immunoglobulin G (IgG), immunoglobulin M (IgM), interleukin-1 β (IL-1β) and interleukin-10 (IL-10) in the serum decreased significantly. (2)Compared with CTX group, the ADG and ADFI of chicks in M. cordata group increased significantly;the F/G decreased significantly;the WBC, red blood cell count (RBC), hemoglobin (HGB), hematokrit (HCT) and mean corpuscular hemoglobin (MCH) increased sig -nificantly;the contents of ALT, AST and MDA in the serum decreased significantly;the contents of TP, GLOB, IgG, Ig M, IL -1β, interleukin-6(IL-6) and IL-10 in the serum increased significantly;and the bursa of fabricius index increased significantly. (3) Compared with control group, the ADG and ADFI of chicks in M. cordata group increased significantly, and the contents of HGB and HCT in the blood increased significantly. [Conclusion] CTX injection will reduce the growth performance, antioxidant and immune function of laying chicks. The addition of M. cordata extract could restore the growth performance, immune function and antioxidant function impaired by CTX.

Effect of Combined Vitamin C and Vitamin E Supplementation on Reproductive Performance and Hatching Rate in Japanese Laying Quail

This study was carried out to evaluate the effect of vitE(Vitamin E)supplementation in combination with vitC(Vitamin C)on reproductive performance and hatching rate of Japanese quail at the age of 59-114 days.A total of 132 laying quails were arranged in a completely randomized design with 3 treatments and 4 replicates,each replicate being 9 female quails and 2 male quails at 59 days of age.The experimental treatments were as follows:control fed the basal diet(KPCS),E125C75 including KPCS supplemented with 125 mg vitE/kg feed combined with 75 mg vitC/kg feed and E125C125 including KPCS supplemented 125 mg vitE/kg feed combined with 125 mg vitC/kg feed.Research results showed that the highest WG(Weight Gain)was in E125C125(30.32 g)and lowest in E125C75(17.37 g).There were no statistically significant differences between treatments in terms of laying rate,feed conversion ratio,egg mass and egg weight.However,hatching rate and bodyweight of quails at 1 day of age were significantly different between treatments,the highest was E125C125(74.37%and 7.43 g),followed by E125C75(70.02%and 7.03 g)and the lowest was control(65.89%and 6.82 g).It can be concluded that E125C125 not only improved laying rate but also increased hatching rate and bodyweight of quail chick.

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.

Hepatic steatosis is associated with dysregulated cholesterol metabolism and altered protein acetylation dynamics in chickens

Background Hepatic steatosis is a prevalent manifestation of fatty liver, that has detrimental effect on the health and productivity of laying hens, resulting in economic losses to the poultry industry. Here, we aimed to systematically investigate the genetic regulatory mechanisms of hepatic steatosis in laying hens.Methods Ninety individuals with the most prominent characteristics were selected from 686 laying hens according to the accumulation of lipid droplets in the liver, and were graded into three groups, including the control, mild hepatic steatosis and severe hepatic steatosis groups. A combination of transcriptome, proteome, acetylome and lipidome analyses, along with bioinformatics analysis were used to screen the key biological processes, modifications and lipids associated with hepatic steatosis.Results The rationality of the hepatic steatosis grouping was verified through liver biochemical assays and RNA-seq. Hepatic steatosis was characterized by increased lipid deposition and multiple metabolic abnormalities. Integration of proteome and acetylome revealed that differentially expressed proteins(DEPs) interacted with differentially acetylated proteins(DAPs) and were involved in maintaining the metabolic balance in the liver. Acetylation alterations mainly occurred in the progression from mild to severe hepatic steatosis, i.e., the enzymes in the fatty acid oxidation and bile acid synthesis pathways were significantly less acetylated in severe hepatic steatosis group than that in mild group(P < 0.05). Lipidomics detected a variety of sphingolipids(SPs) and glycerophospholipids(GPs) were negatively correlated with hepatic steatosis(r ≤-0.5, P < 0.05). Furthermore, the severity of hepatic steatosis was associated with a decrease in cholesterol and bile acid synthesis and an increase in exogenous cholesterol transport.Conclusions In addition to acquiring a global and thorough picture of hepatic steatosis in laying hens, we were able to reveal the role of acetylation in hepatic steatosis and depict the changes in hepatic cholesterol metabolism. The findings provides a wealth of information to facilitate a deeper understanding of the pathophysiology of fatty liver and contributes to the development of therapeutic strategies.

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.

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.

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.