Background Dietary supplementation of xylooligosac charides(XOS) has been found to influence gut health by manipulating cecal microbiota and producing microbe-origin metabolites.But no study investigated and compared ...Background Dietary supplementation of xylooligosac charides(XOS) has been found to influence gut health by manipulating cecal microbiota and producing microbe-origin metabolites.But no study investigated and compared the effect of in ovo feeding of xylobiose(XOS2) and xy lotriose(XOS3) in chickens.This study investigated the effect of in ovo feeding of these XOS compounds on post-hatch gut health parameters in chickens.A total of 144 fertilized chicken eggs were divided into three groups:a) non-injected control(CON),b) XOS2,and c) XOS3.On the 17^(th) embryonic day,the eggs of the XOS2 and XOS3 groups were injected with 3 mg of XOS2 and XOS3 diluted in 0.5 mL of 0.85% normal saline through the amniotic sac.After hatching,the chicks were raised for 21 d.Blood was collected on d 14 to measure plasma immunoglobulin.Cecal digests were collected for measuring short-chain fatty acids(SCFA) on d 14 and 21,and for microbial ecology and microbial metabolic pathway analyses on d 7 and 21.Results The results were considered significantly different at P<0.05.ELISA quantified plasma IgA and IgG on d 14chickens,revealing no differences among the treatments.Gas chromatography results showed no significant differences in the concentrations of cecal SCFAs on d 14 but significant differences on d 21.However;the SCFA concentrations were lower in the XOS3 than in the CON group on d 21.The cecal metagenomics data showed that the abundance of the family Clostridiaceae significantly decreased on d 7,and the abundance of the family Oscillospiraceae increased on d 21 in the XOS2 compared to the CON.There was a reduction in the relative abundance of genus Clostridium sense stricto 1 in the XOS2 compared to the CON on d 7 and the genus Ruminococcus,torques in both XOS2 and XOS3 groups compared to the CON on d 21.The XOS2 and XOS3 groups reduced the genes for chondroitin sulfate degradation Ⅰ and L-histidine degradation Ⅰpathways,which contribute to improved gut health,respectivelyc in the microbiome on d 7.In contrast,on d 21,the XOS2 and XOS3 groups enriched the thiamin salvage Ⅱ,L-isoleucine biosynthesis Ⅳ,and O-antigen building blocks biosynthesis(E. coli) pathways,which are indicative of improved gut health.Unlike the XOS3 and CON,the microbiome enriched the pathways associated with energy enhancement,including flavin biosynthesis Ⅰ,sucrose degradation Ⅲ,and Calvin-Benson-Bassham cycle pathways,in the XOS2 group on d 21.Conclusion In ovo XOS2 and XOS3 feeding promoted beneficial bacterial growth and reduced harmful bacteria at the family and genus levels.The metagenomic-based microbial metabolic pathway profiling predicted a favorable change in the availability of cecal metabolites in the XOS2 and XOS3 groups.The modulation of microbiota and metabolic pathways suggests that in ovo XOS2 and XOS3 feeding improved gut health during the post-hatch period of broilers.展开更多
Background:Prenatal nutrition is crucial for embryonic development and neonatal growth,and has the potential to be a main determinant of life-long health.In the present study,we used a layer chick model to investigate...Background:Prenatal nutrition is crucial for embryonic development and neonatal growth,and has the potential to be a main determinant of life-long health.In the present study,we used a layer chick model to investigate the effects of in ovo feeding(IOF)of L-arginine(Arg)on growth,intestinal development,intestinal microbiota and metabolism.The treatments included the non-injected control,saline-injected control,and saline containing 2,6,or 10 mg Arg groups.Results:IOF Arg increased early intestinal index and villus height,and enhanced uptake of residual yolk lipid,contributing to subsequent improvement in the early growth performance of chicks.Prenatal Arg supplementation also increased the early microbialα-diversity,the relative abundance of Lactobacillales and Clostridiales,and decreased the relative abundance of Proteobacteria of cecum in chicks.Furthermore,the shift of cecal microbiota composition and the colonization of potential probiotics were accelerated by IOF of Arg.Simultaneously,metabolomics showed that metabolisms of galactose,taurine-conjugated bile acids and lipids were modulated to direct more energy and nutrients towards rapid growth of intestine at the beginning of post-hatch when embryos received IOF of Arg.Conclusions:Prenatal Arg supplementation showed beneficial effects on the early intestinal development,cecal microbiota and host metabolism of layer chicks,contributing to subsequent improvement in the early growth performance.These findings provide new insight into the role of IOF of Arg in the establishment of the gut microbiota of newly-hatched layer chicks,and can expand our fundamental knowledge about prenatal nutrition,early bacterial colonization and intestinal development in neonate.展开更多
Background:This study investigated a novel in ovo feeding strategy to determine the prebiotic effects of xylo-and mannan-oligosaccharides(XOS and MOS)differing in the degree of polymerization.A total of 192 fertilized...Background:This study investigated a novel in ovo feeding strategy to determine the prebiotic effects of xylo-and mannan-oligosaccharides(XOS and MOS)differing in the degree of polymerization.A total of 192 fertilized eggs were divided into 6 treatment groups:i)normal saline control(NSC),ii)xylotriose(XOS3),iii)xylotetraose(XOS4),iv)mannotriose(MOS3),v)mannotetraose(MOS4),and vi)no injection control(NIC),each containing 4 replicate trays with 8 eggs per replicate.On d 17 of incubation,3 mg of oligosaccharides(except for controls)dissolved in 0.5 mL of 0.85%normal saline were injected into the amnion of Cobb 500 broilers eggs.After hatch,the chicks were raised for 28 d under standard husbandry practices and were fed a commercial broilers diet ad libitum,and samples were collected periodically.Results:The hatchability,growth performance,and relative weights of breast,drumstick,liver,and proventriculus were not different among the treatments(P>0.05).The XOS3 injection increased the total short-chain fatty acid production at d 28 compared with both control groups(P<0.05).The villus height to crypt depth ratio was significantly higher in the XOS4 group than both controls on the hatch day(P<0.01)but were not different among any treatments on d 7 and 28(P>0.05).On the hatch day,the expression level of the CD3 gene(a T cell marker)was increased by XOS3,while the IL-10 gene(a marker of anti-inflammatory cytokine)was reduced by MOS4(P<0.05)compared with both controls.Compared with both controls,XOS3 exhibited a trend of reduction for IL-10(P=0.074).No cytokines or lymphocyte markers were affected by the treatments on d 7(P>0.05),except XOS4 increased IL-4 compared with NSC(P<0.05).The broilers in the MOS4 group had higher operational taxonomic units(OTUs)and had more differentially abundant taxa,including order Lactobacillales and family Leuconostocaceae(P<0.05)than both controls on d 28.The predictive functional profiling indicated that the linoleic acid metabolism pathway was enriched in the cecal microbiota of the XOS3 group compared with both controls(P<0.05).Conclusion:The effects of these XOS and MOS on ileal mucosa and immunity are transient,but the effects on fermentation and cecal microbiota are prolonged,and further research is warranted to determine their use as a gut health promoter in poultry.展开更多
Two experiments were conducted to investigate the effect of in ovo zinc （Zn） injection on the embryonic development, tissue Zn contents, antioxidation and related gene expressions of fertilized eggs of Arbor Acres b...Two experiments were conducted to investigate the effect of in ovo zinc （Zn） injection on the embryonic development, tissue Zn contents, antioxidation and related gene expressions of fertilized eggs of Arbor Acres broiler breeders. Experiment 1 was conducted to determine an optimal embryonic age for early in ovo injection. A total of 720 fertilized eggs with similar weights were randomly allotted to 4 treatments with 6 replicates per treatment and 30 eggs per replicate in a completely randomized design. The eggs were injected with 0.1 mL sterilized water at 3, 6 and 9 embryonic days of incubation （E3, E6 and E9） or non-injection （the control）, respectively. The results from experiment 1 showed that E3 and E6 injections increased （P〈0.05） the embryonic mortalities, and decreased （P〈0.05） hatchabilities compared to the non-injected control, but no differences （P〉0.05） between E9 injection and the non-injected control were observed in either embryonic mortality or hatchability. The findings suggest that the E9 is the optimal embryonic age for early in ovo injection. In experiment 2, a total of 672 fertilized eggs with similar weights were randomly allocated to 7 treatments with 6 replicates per treatment and 16 eggs per replicate in a completely randomized design. The eggs were injected with 0 （the negative control）, 50, 100, 150, 200, or 250 μg Zn/egg as reagent grade ZnSO4.7H20 in a 0.1-mL solution, or non-injection （the positive control）, respectively at E9-10. The results from the experiment 2 demonstrated that no differences （P〉0.05） among 50 and 100 μg Zn/egg groups and the negative control were observed in the embryonic mortality and hatchability, however, the injection of 200 μg Zn/egg increased （P〈0.05） the embryonic mortality, and injections of 150 and 200 μg Zn/egg decreased （P〈0.05） hatchabilities compared with the controls. The embryonic tibia Zn contents at E20 were increased （P〈0.05） by injections of 150, 200 and 250μg Zn/egg. Zinc injection did not affect （P〉0.05） malonaldehyde （MDA） contents, copper- and Zn- containing superoxide dismutase （CuZnSOD） activities and mRNA expression levels in the liver and heart of chick embryos at E15 and E20. Compared with the negative control, injections of 50, 150 and 200 μg Zn/egg up-regulated （P〈0.05） themetallothionein （MT） mRNA expression levels in the embryonic liver at E20. These results indicated that in ovo Zn injections increased Zn contents in the embryonic tibia and MT mRNA expression levels in the embryonic liver at E20, however, injections of 150-200 μg Zn/egg were harmful to the embryonic development.展开更多
Background： Application the innovative method which is in ovo technology provides a means of modulating the immune system at early embryonic stages. The aim of study was to determine influence of the in ovo stimulati...Background： Application the innovative method which is in ovo technology provides a means of modulating the immune system at early embryonic stages. The aim of study was to determine influence of the in ovo stimulation, on d12 of incubation, with synbiotics（synbiotic 1-L. salivarius IBB3154 ＋ Bi^2tos, Clasado Ltd. and the synbiotic 2-L. plantarum IBB3036 ＋ lupin RFOs） on the microstructure of duodenum, jejunum and ileum in the 1^（st） and 42^（nd）day of rearing.Results： On the 1^（st） day of chickens life, in the duodenum of both experimental groups（SYN1 and SYN2）, a significantly higher and wider intestinal vil i as wel as a significantly larger absorbent surface of these villi were found in comparison with the Control group（P ≤ 0.01）. On the 42^（nd）day of rearing the beneficial effect of synbiotic 1 was reflected by the numerical y higher vil i（no statistical differences） with a larger surface（P ≤ 0.01） in the duodenum in the SYN1 group compare to the Control group. In the jejunum on the 1^（st） day of life, in the SYN1 group, significantly higher vil i than in the Control group, with a simultaneous decrease in the depth of crypts（P ≤ 0.01）, and also the largest width of vil i and their absorbent area（P ≤ 0.01） in comparison to the other groups were found. On the 42^（nd）day of life, in the jejunum, an increase in the height of the vil i whilst reducing the crypt depth in the SYN2 group was found（P ≤ 0.01）. In turn, in the SYN1 group, there were significantly more neutral goblet cel s observed compared with the control group（P ≤ 0.05）. In the ileum of 1-day-old chickens, the widest vil i（P ≤ 0.05） and the deepest crypts（P ≤ 0.01） were found in the SYN2 group. In the same group, there was also the least amount of neutral goblet cel s in comparison to the other groups（P ≤ 0.05）.Conclusions： We observed that synbiotic 1 and 2 beneficial y affected the examined characteristics on the 1^（st） and 42^（nd） day of life. The obtained results al ow us to conclude that the use of synbiotics significantly affect gut structure which should contribute to improvement in nutrient absorption by the gut.展开更多
Background： A leptin-like immunoreactive substance has been found in chicken eggs and has been implicated in serving as a maternal signal to program offspring growth and metabolism. In the present study, we investiga...Background： A leptin-like immunoreactive substance has been found in chicken eggs and has been implicated in serving as a maternal signal to program offspring growth and metabolism. In the present study, we investigated the effects of in ovo leptin administration on hatch weight, serum and hepatic concentrations of metabolites and hormones, as well as on the expression of genes involved in hepatic lipid metabolism and the predicted microRNAs （miRNAs） targeting the affected genes. To this end we injected fertile eggs with either 0.5 μg of recombinant murine leptin or vehicle （PBS） before incubation.Results： Prenatally leptin-exposed chicks showed lower hatch weight, but higher liver weight relative to the body weight, compared to the control group. In ovo leptin treatment increased the hepatic content and serum concentration of leptin in newly hatched chickens. The hepatic contents of triglycerides （TG） and total cholesterol （Tch） were decreased, whereas the serum levels of TG, Tch and apolipoprotein B （ApoB） were increased. The hepatic mRNA expression of sterol regulator element binding protein 1 （SREBP-1c）, SREBP-2, hydroxy-3-methylglutaryl coenzyme A reductase （HMGCR） and cholesterol 7a-hydroxylase 1 （CYP7A1） was significantly up-regulated, as was the protein content of both SREBP-1 c and SREBP-2 in hepatic nuclear extracts of leptin-treated chickens. Moreover, out of 12 miRNAs targeting SREBP-1c and/or HMGCR, five were significantly up-regulated in liver of leptin-treated chicks, including gga-miR-200b and gga-miR-429, which target both SREBP-1c and HMGCR.Conclusions： These results suggest that leptin in ovo decreases hatch weight, and modifies hepatic leptin secretion and lipid metabolism in newly hatched broiler chickens, possibly via microRNA-mediated gene regulation.展开更多
Numerous researchers and institutions have been developing in ovo sexing technologies to improve animal welfare by identifying male embryos in an early embryonic stage and disposing of them before pain perception.This...Numerous researchers and institutions have been developing in ovo sexing technologies to improve animal welfare by identifying male embryos in an early embryonic stage and disposing of them before pain perception.This review gives a complete overview of the technological approaches reported in papers and patents by performing a thorough search using Web of Science and Patstat/Espacenet databases for papers and patents,respectively.Based on a total of 49 papers and 115 patent families reported until May 2023 worldwide,11 technology categories were defined:6 non-optical and 5 optical techniques.Every category was described for its characteristics while assessing its potential for application.Next,the dynamics of the publications of in ovo sexing techniques in both paper and patent fields were described through growth curves,and the interest or actual status was visualized using the number of paper citations and the actual legal status of the patents.When comparing the reported technologies in papers to those in patents,scientific gaps were observed,as some of the patented technologies were not reported in the scientific literature,e.g.,ion mobility and mass spectrometry approaches.Generally,more diverse approaches in all categories were found in patents,although they do require more scientific evidence through papers or industrial adoption to prove their robustness.Moreover,although there is a recent trend for non-invasive techniques,invasive methods like analyzing DNA through PCR or hormones through immunosensing are still being reported(and might continue to be)in papers and patents.It was also observed that none of the technologies complies with all the industry requirements,although 5 companies already entered the market.On the one hand,more research and harmony between consumers,industry,and governments is necessary.On the other hand,close monitoring of the market performance of the currently available techniques will offer valuable insights into the potential and expectations of in ovo sexing techniques in the poultry industry.展开更多
Background:Many researches about in ovo feeding(IOF)of vitamin C(VC)are gradually carried out to explore physiological development in chicken,but little studies focus on VC synthesis capacity of the embryo itself,the ...Background:Many researches about in ovo feeding(IOF)of vitamin C(VC)are gradually carried out to explore physiological development in chicken,but little studies focus on VC synthesis capacity of the embryo itself,the selection of injection site and the effectiveness of IOF of VC.This study aims to explore the above problems.Results:Kidney and yolk sac were the main organs for VC synthesis and L-gulonolactone oxidase(GLO)expression was lower during pre-hatch development than that during post-hatch development.Sodium-dependent vitamin C transporter 1(SVCT1)expression was increased continuously in yolk sac from embryonic age 19(E19)to post-hatch day 1(D1)and in intestine(duodenum,jejunum and ileum)from E17 to D1.Plasma VC content was higher at D1 than that at D21 and D42.IOF of VC significantly reduced GLO expression in liver,kidney and yolk sac as well as SVCT1 expression in duodenum,jejunum and ileum,but increased the VC content in plasma,brain,kidney and liver.In addition,IOF of VC obviously reduced the embryonic morality and increased the hatchability under heat stress.Conclusions:This study suggested that IOF of VC at E11 in yolk was effective for embryonic VC supplementation.These findings provide a theoretical reference about the method of embryonic VC supplementation and effective methodology on embryonic VC nutrition in broiler chickens.展开更多
Background Broiler chickens are most vulnerable immediately after hatching due to their immature immune systems,making them susceptible to infectious diseases.The yolk plays an important role in early immune defence b...Background Broiler chickens are most vulnerable immediately after hatching due to their immature immune systems,making them susceptible to infectious diseases.The yolk plays an important role in early immune defence by showing relevant antioxidant and passive immunity capabilities during broiler embryonic development.The immunomodulatory effects of phytogenic compound carvacrol have been widely reported.After in ovo delivery in the amniotic fluid during embryonic development carvacrol is known to migrate to the yolk sac.However,it is unknown whether carvacrol in the yolk could enhance defence responsiveness in the yolk sac.Therefore,the aim of this study was to improve early immune function in chicken embryos,and it was hypothesized that in ovo delivery of carvacrol would result in immunomodulatory effects in the yolk sac,potentially improving post-hatch resilience.Methods On embryonic day(E)17.5,either a saline(control)or carvacrol solution was injected into the amniotic fluid.Yolk sac tissue samples were collected at E19.5,and transcriptomic analyses using RNA sequencing were performed,following functional enrichment analyses comparing the control(saline)and carvacrol-injected groups.Results The results showed that 268 genes were upregulated and 174 downregulated in the carvacrol group compared to the control(P<0.05;logFC<-0.5 or log FC>0.5).Functional analyses of these differentially expressed genes,using KEGG,REACTOME,and Gene Ontology databases,showed enrichment of several immune-related pathways.This included the pathways‘Antimicrobial peptides’(P=0.001)and‘Chemoattractant activity’(P=0.004),amongst others.Moreover,the‘NOD-like receptor signaling’pathway was enriched(P=0.002).Antimicrobial peptides are part of the innate immune defence and are amongst the molecules produced after the nucleotide oligomeriza-tion domain(NOD)-like receptor pathway activation.While these responses may be associated with an inflammatory reaction to an exogenous threat,they could also indicate that in ovo delivery of carvacrol could prepare the newly hatched chick against bacterial pathogens by potentially promoting antimicrobial peptide production through acti-vation of NOD-like receptor signaling in the yolk sac.Conclusion In conclusion,these findings suggest that in ovo delivery of carvacrol has the potential to enhance anti-pathogenic and pro-inflammatory responses in the yolk sac via upregulation of antimicrobial peptides,and NOD-like receptor pathways.展开更多
BACKGROUND Cardiovascular diseases are the first cause of death in the world.Ischemic heart disease is the main cause of heart failure.New approaches are continuously sou-ght to identify better therapeutic success.The...BACKGROUND Cardiovascular diseases are the first cause of death in the world.Ischemic heart disease is the main cause of heart failure.New approaches are continuously sou-ght to identify better therapeutic success.Thereby,current research has been drawn to identifying and completing the therapeutic profile of natural sources.Galium species are representatives exhibiting diuretic and antibacterial potential in living organisms and can treat burns,wounds,and skin diseases.Moreover,it was also observed that these plants manifest cardioprotective effects as well as having antihemolytic,antioxidant,antibacterial,anti-inflammatory,immunomod-ulatory,and antiproliferative potential.In ischemic heart disease,Galium verum(G.verum)extract manifested preservative properties in terms of contractility,systolic and diastolic function maintenance,and reduced damage to the heart after ischemia.In addition,G.verum extract upregulated the activity of antioxidant enzymes alleviating the production of pro-oxidants.AIM To test the ethanolic extract of G.verum on the H9C2(2-1)cell line by evaluating the in vitro biosafety profile and in ovo irritative potential.METHODS Cells were tested in vitro for viability(using the MTT test),cellular morphology,cell number,confluence,nuclear morphology(by immunofluorescence staining of cell nuclei and F-actin assay)and in ovo by the hen’s egg chorioal-lantoic membrane(CAM)test and CAM anti-irritant methods to study the irritation potential on the CAM.RESULTS The extract demonstrated a dose-dependent stimulatory activity.The viability increased to 170%for the dose of 55µg/mL and decreased to 135%at 200µg/mL.The results of cell number,confluence,and morphological analysis did not present significant changes compared with control untreated cells.The immunofluorescence assay showed insignificant apoptotic potential,and the hen’s egg CAM test revealed that the extract was in the weak to mode-rately irritating category with an irritation score of 5.3.When applying the sample to the CAM,only slight coagu-lation was observed(128 s).The anti-irritant test revealed the protective potential of the extract in the vascular plexus.CONCLUSION The ethanolic extract of G.verum manifests a stimulating effect on cardiomyocytes,enhancing cell viability,and maintaining a normal elongated shape,cell number,and confluence,without significant signs of apoptosis and with a weak irritative effect in ovo.In addition,the extract demonstrated a protective effect against hemorrhage,lysis,and coagulation of blood vessels induced by sodium dodecyl sulfate on the CAM.展开更多
Background Hatch weight(HW)affects broiler growth and low HW(LHW)often leads to suboptimal performance.Sodium butyrate(SB)has been shown to promote growth through enhanced intestinal health.This study investi-gated ho...Background Hatch weight(HW)affects broiler growth and low HW(LHW)often leads to suboptimal performance.Sodium butyrate(SB)has been shown to promote growth through enhanced intestinal health.This study investi-gated how broilers with different HW responded to in ovo SB injection and whether SB could enhance gut health and performance in LHW chicks.Ross 308 broiler eggs were injected on incubation d 12 with physiological saline(control)or SB at 0.1%(SB1),0.3%(SB3),or 0.5%(SB5).Post-hatch,male chicks from each treatment were categorized as high HW(HHW)or LHW and assigned to 8 groups in a 4×2 factorial design.Production parameters were recorded periodically.Intestinal weight,length,and gene expression related to gut barrier function and immune response were examined on d 14 and 42.Cecal microbiota dynamics and predicted functionality were analyzed using 16S rRNA gene sequencing.Results SB treatments did not affect hatchability.HHW-control group exhibited consistently better weight gain and FCR than LHW-control group.SB dose-dependently influenced performance and gut health in both HW catego-ries,with greater effects in LHW broilers at 0.3%.LHW-SB3 group attained highest body weight on d 42,exceeding controls but not significantly differing from HHW-SB3 group.LHW-SB3 group showed upregulation of gut-barrier genes CLDN1 in ileum,TJP1 in jejunum and anti-inflammatory cytokine IL-10 in both jejunum and ileum on d 14.Addi-tionally,LHW-SB3 group upregulated mucin-producing MUC6 gene in ileum,while HHW-SB5 group increased pro-inflammatory IL-12p40 cytokine in caecum on d 42.LHW-SB3 group demonstrated shorter relative intestinal lengths,while HHW-SB5 had longer lengths.HHW-control group had higher bacterial diversity and growth-promoting bacte-ria while LHW-control group harbored the potential pathogen Helicobacter.SB reshaped gut microbiota biodiversity,composition,and predicted metabolic pathways in both HW categories.The LHW-SB3 group exhibited highest alpha diversity on d 14 and most beneficial bacteria at all timepoints.HHW-SB5 group presented increased pathogenic Escherichia-Shigella and Campylobacter on d 42.Conclusions HW significantly affects subsequent performance and SB has differential effects based on HW.LHW chicks benefited more from 0.3%SB,showing improvements in growth,intestinal development,health,and gut microbiota characteristics.展开更多
Selenium is an essential trace element for humans and animals.As the active center of selenoproteins,the addition of selenium is beneficial to enhance the antioxidant ability.However,the high cost limits the applicati...Selenium is an essential trace element for humans and animals.As the active center of selenoproteins,the addition of selenium is beneficial to enhance the antioxidant ability.However,the high cost limits the application of organic Se in agriculture animal production.Selenized glucose(SeGlu)is a newly invented organoselenium material with good stability,low toxicity and low cost.This assay found that SeGlu was able to increase selenium deposition in liver of newborn broilers,and enhance the antioxidant capacity of liver by elevating the activities of antioxidant enzymes such as total superoxide dismutase and glutathione peroxidase.This paper as the first example clarifying the mechanism of SeGlu to enhance the antioxidant ability of chicks,shows that SeGlu can be used as an organic selenium enrichment additive for early nutrition of poultry.As a cross-discipline study involving chemistry,biology and agriculture animal science,the work may be beneficial for studies in related fields and prompt the development of the selenium science.展开更多
The intestinal health of chick embryos is vital for their life-long growth,and exogenous nutrition intervention may provide sufficient nutrition for embryonic development.In the present study,we investigated the effec...The intestinal health of chick embryos is vital for their life-long growth,and exogenous nutrition intervention may provide sufficient nutrition for embryonic development.In the present study,we investigated the effect of in ovo injection of L-methionine(L-Met)on the intestinal structure and barrier function of chick embryos.There were 4 groups of treatments:the control(CON)group injected with phosphate-buffered saline(PBS)and the other 3 groups injected with 5,10,and 20 mg L-Met/egg,respectively.The injection was performed on embryonic day 9(E9),and intestinal samples were collected on the day of hatching for analysis.The results showed that,compared with the CON group,the groups administered an in ovo injection of L-Met increased relative weights of the duodenum,jejunum,and ileum(P<0.05).Hematoxylin and eosin(H&E)staining showed that the groups injected with 5,10,and 20 mg L-Met significantly increased villus height and crypt depth(P<0.05).Moreover,in ovo injection of 10 mg L-Met also increased the transepithelial electrical resistance(TEER)of the jejunum(P<0.05).Injection with 10 and 20 mg L-Met increased the expression of the tight junction proteins(ZO-1 and claudin-1)and the fluorescence signal intensity of Ki67 and villin proteins(P<0.05).Further,the protein expression of phospho-Janus kinase 2(p-JAK2)and phospho-signal transducer and activator of transcription 3(p-STAT3)was significantly increased by 10 or 20 mg L-Met injection(P<0.05).In conclusion,the injection of L-Met,especially at a dose of 10 mg,showed beneficial effects on the intestinal integrity of chick embryos due to the activation of the JAK2/STAT3 signaling pathway.Our results may provide new insights for regulating the intestinal development of embryonic chicks and the rapid growth of chicks after hatching.展开更多
Healthy chickens are necessary to meet the ever-increasing demand for poultry meat.Birds are subjected to numerous stressful conditions under commercial rearing systems,including variations in the envi-ronmental tempe...Healthy chickens are necessary to meet the ever-increasing demand for poultry meat.Birds are subjected to numerous stressful conditions under commercial rearing systems,including variations in the envi-ronmental temperature.However,it is difficult to counter the effects of global warming on the livestock industry.High environmental temperature is a stressful condition that has detrimental effects on growth and production performance,resulting in decreased feed intake,retarded growth,compromised gut health,enhanced oxidative stress,and altered immune responses.Traditional approaches include nutritional modification and housing management to mitigate the harmful effects of hot environments.Currently,broiler chickens are more susceptible to heat stress(HS)than layer chickens because of their high muscle mass and metabolic rate.In this review,we explored the possibility of in ovo manipulation to combat HS in broiler chickens.Given their short lifespan from hatching to market age,embryonic life is thought to be one of the critical periods for achieving these objectives.Chicken embryos can be modulated through either temperature treatment or nourishment to improve thermal tolerance during the rearing phase.We first provided a brief overview of the harmful effects of HS on poultry.An in-depth evaluation was then presented for in ovo feeding and thermal manipulation as emerging strategies to combat the negative effects of HS.Finally,we evaluated a combination of the two methods using the available data.Taken together,these investigations suggest that embryonic manipulation has the po-tential to confer heat resistance in chickens.展开更多
Members of the basic helix-loop-helix （bHLH） gene family play important roles in vertebrate neurogenesis. In this study, confocal microscopy-based fluorescence resonance energy transfer （FRET） is used to monitor b...Members of the basic helix-loop-helix （bHLH） gene family play important roles in vertebrate neurogenesis. In this study, confocal microscopy-based fluorescence resonance energy transfer （FRET） is used to monitor bHLH protein-protein interactions under various physiological conditions. Tissue-specific bHLH activators, NeuroD 1, Mash 1, Neurogenin 1 （Ngn 1）, Neurogenin2 （Ngn2）, and ubiquitous expressed E47 protein are tagged with enhanced yellow fluorescence protein （EYFP） and enhanced cyan fluorescence protein （ECFP）, respectively. The subcellular localization and mobility ofbHLH fusion proteins are examined in HEK293 cells. By transient transfection and in ovo electroporation, four pairs of tissue-specific bHLH activators and E47 protein are over-expressed in HEK293 cells and developing chick embryo neural tube. With the acceptor photobleaching method, FRET could be detected between these bHLH protein pairs in the nuclei of transfected cells and developing neural tubes. Mashl/E47 and Ngn2/E47 FRET pairs show higher FRET efficiencies in the medial and the lateral half of chick embryo neural tube, respectively. It suggests that these bHLH protein pairs formed functional DNA-protein complexes with regulatory elements of their downstream target genes in the specific regions. This work will help one understand the behaviours of bHLH factors in vivo.展开更多
Methionine(Met)is an essential and first limiting amino acid in the poultry diet that plays a significant role in chicken embryonic development and growth.The present study examined the effect of in ovo injection of D...Methionine(Met)is an essential and first limiting amino acid in the poultry diet that plays a significant role in chicken embryonic development and growth.The present study examined the effect of in ovo injection of DL-Met and L-Met sources and genotypes on chicken embryonic-intestinal development and health.Fertilized eggs of the two genotypes,TETRA-SL layer hybrid(TSL)—commercial layer hybrid and Hungarian Partridge colored hen breed(HPC)—a native genotype,were randomly distributed into four treatments for each genotype.The treatment groups include the following:1)control non-injected eggs(NoIn);2)saline-injected(SaIn);3)DL-Met injected(DLM);and 4)L-Met injected(LM).The in ovo injection was carried out on 17.5 d of embryonic development;after hatching,eight chicks per group were sacrificed,and the jejunum was extracted for analysis.The results showed that both DLM and LM groups had enhanced intestinal development as evidenced by increased villus width,villus height,and villus area(P<0.05)compared to the control.The DLM group had significantly reduced crypt depth,glutathione(GSH)content,glutathione S-transferase 3 alpha(GST3),occludin(OCLN)gene expression and increased villus height to crypt depth ratio in the TSL genotype than the LM group(P<0.05).The HPC genotype has overexpressed insulin-like growth factor 1(IGF1)gene,tricellulin(MD2),occludin(OCLN),superoxide dismutase 1(SOD1),and GST3 genes than the TSL genotype(P<0.05).In conclusion,these findings showed that in ovo injection of Met enhanced intestinal development,and function,with genotypes responding differently under normal conditions.Genotypes also influenced the expression of intestinal antioxidants,tight junction,and growth-related genes.展开更多
基金supported by the USDA National Institute for Food and Agriculture,Hatch-Multistate Fund,managed by the College of Tropical Agriculture and Human Resources,University of Hawaii at Manoa,Honolulu,HI,USA。
文摘Background Dietary supplementation of xylooligosac charides(XOS) has been found to influence gut health by manipulating cecal microbiota and producing microbe-origin metabolites.But no study investigated and compared the effect of in ovo feeding of xylobiose(XOS2) and xy lotriose(XOS3) in chickens.This study investigated the effect of in ovo feeding of these XOS compounds on post-hatch gut health parameters in chickens.A total of 144 fertilized chicken eggs were divided into three groups:a) non-injected control(CON),b) XOS2,and c) XOS3.On the 17^(th) embryonic day,the eggs of the XOS2 and XOS3 groups were injected with 3 mg of XOS2 and XOS3 diluted in 0.5 mL of 0.85% normal saline through the amniotic sac.After hatching,the chicks were raised for 21 d.Blood was collected on d 14 to measure plasma immunoglobulin.Cecal digests were collected for measuring short-chain fatty acids(SCFA) on d 14 and 21,and for microbial ecology and microbial metabolic pathway analyses on d 7 and 21.Results The results were considered significantly different at P<0.05.ELISA quantified plasma IgA and IgG on d 14chickens,revealing no differences among the treatments.Gas chromatography results showed no significant differences in the concentrations of cecal SCFAs on d 14 but significant differences on d 21.However;the SCFA concentrations were lower in the XOS3 than in the CON group on d 21.The cecal metagenomics data showed that the abundance of the family Clostridiaceae significantly decreased on d 7,and the abundance of the family Oscillospiraceae increased on d 21 in the XOS2 compared to the CON.There was a reduction in the relative abundance of genus Clostridium sense stricto 1 in the XOS2 compared to the CON on d 7 and the genus Ruminococcus,torques in both XOS2 and XOS3 groups compared to the CON on d 21.The XOS2 and XOS3 groups reduced the genes for chondroitin sulfate degradation Ⅰ and L-histidine degradation Ⅰpathways,which contribute to improved gut health,respectivelyc in the microbiome on d 7.In contrast,on d 21,the XOS2 and XOS3 groups enriched the thiamin salvage Ⅱ,L-isoleucine biosynthesis Ⅳ,and O-antigen building blocks biosynthesis(E. coli) pathways,which are indicative of improved gut health.Unlike the XOS3 and CON,the microbiome enriched the pathways associated with energy enhancement,including flavin biosynthesis Ⅰ,sucrose degradation Ⅲ,and Calvin-Benson-Bassham cycle pathways,in the XOS2 group on d 21.Conclusion In ovo XOS2 and XOS3 feeding promoted beneficial bacterial growth and reduced harmful bacteria at the family and genus levels.The metagenomic-based microbial metabolic pathway profiling predicted a favorable change in the availability of cecal metabolites in the XOS2 and XOS3 groups.The modulation of microbiota and metabolic pathways suggests that in ovo XOS2 and XOS3 feeding improved gut health during the post-hatch period of broilers.
基金This study was supported by National Key R&D Program of China(2017YFD0500500)the earmarked fund for Modern Agro-industry Technology Research System(CARS-40-K12)+1 种基金Beijing Innovation Consortium of Agriculture Research System(BAIC04–2018)the Agricultural Science and Technology Innovation Program(ASTIP)of the Chinese Academy of Agricultural Sciences.
文摘Background:Prenatal nutrition is crucial for embryonic development and neonatal growth,and has the potential to be a main determinant of life-long health.In the present study,we used a layer chick model to investigate the effects of in ovo feeding(IOF)of L-arginine(Arg)on growth,intestinal development,intestinal microbiota and metabolism.The treatments included the non-injected control,saline-injected control,and saline containing 2,6,or 10 mg Arg groups.Results:IOF Arg increased early intestinal index and villus height,and enhanced uptake of residual yolk lipid,contributing to subsequent improvement in the early growth performance of chicks.Prenatal Arg supplementation also increased the early microbialα-diversity,the relative abundance of Lactobacillales and Clostridiales,and decreased the relative abundance of Proteobacteria of cecum in chicks.Furthermore,the shift of cecal microbiota composition and the colonization of potential probiotics were accelerated by IOF of Arg.Simultaneously,metabolomics showed that metabolisms of galactose,taurine-conjugated bile acids and lipids were modulated to direct more energy and nutrients towards rapid growth of intestine at the beginning of post-hatch when embryos received IOF of Arg.Conclusions:Prenatal Arg supplementation showed beneficial effects on the early intestinal development,cecal microbiota and host metabolism of layer chicks,contributing to subsequent improvement in the early growth performance.These findings provide new insight into the role of IOF of Arg in the establishment of the gut microbiota of newly-hatched layer chicks,and can expand our fundamental knowledge about prenatal nutrition,early bacterial colonization and intestinal development in neonate.
基金supported by the USDA National Institute for Food and Agriculture,Hatch-Multistate Fundby the College of Tropical Agriculture and Human Resources,University of Hawaii at Manoa,Honolulu,HI,USA
文摘Background:This study investigated a novel in ovo feeding strategy to determine the prebiotic effects of xylo-and mannan-oligosaccharides(XOS and MOS)differing in the degree of polymerization.A total of 192 fertilized eggs were divided into 6 treatment groups:i)normal saline control(NSC),ii)xylotriose(XOS3),iii)xylotetraose(XOS4),iv)mannotriose(MOS3),v)mannotetraose(MOS4),and vi)no injection control(NIC),each containing 4 replicate trays with 8 eggs per replicate.On d 17 of incubation,3 mg of oligosaccharides(except for controls)dissolved in 0.5 mL of 0.85%normal saline were injected into the amnion of Cobb 500 broilers eggs.After hatch,the chicks were raised for 28 d under standard husbandry practices and were fed a commercial broilers diet ad libitum,and samples were collected periodically.Results:The hatchability,growth performance,and relative weights of breast,drumstick,liver,and proventriculus were not different among the treatments(P>0.05).The XOS3 injection increased the total short-chain fatty acid production at d 28 compared with both control groups(P<0.05).The villus height to crypt depth ratio was significantly higher in the XOS4 group than both controls on the hatch day(P<0.01)but were not different among any treatments on d 7 and 28(P>0.05).On the hatch day,the expression level of the CD3 gene(a T cell marker)was increased by XOS3,while the IL-10 gene(a marker of anti-inflammatory cytokine)was reduced by MOS4(P<0.05)compared with both controls.Compared with both controls,XOS3 exhibited a trend of reduction for IL-10(P=0.074).No cytokines or lymphocyte markers were affected by the treatments on d 7(P>0.05),except XOS4 increased IL-4 compared with NSC(P<0.05).The broilers in the MOS4 group had higher operational taxonomic units(OTUs)and had more differentially abundant taxa,including order Lactobacillales and family Leuconostocaceae(P<0.05)than both controls on d 28.The predictive functional profiling indicated that the linoleic acid metabolism pathway was enriched in the cecal microbiota of the XOS3 group compared with both controls(P<0.05).Conclusion:The effects of these XOS and MOS on ileal mucosa and immunity are transient,but the effects on fermentation and cecal microbiota are prolonged,and further research is warranted to determine their use as a gut health promoter in poultry.
基金supported by the Key International Cooperation Program of the National Natural Science Foundation of China (31110103916)the Agricultural Science and Technology Innovation Program, China (ASTIPIAS08)the earmaked fund for the China Agriculture Research System (CARS-42)
文摘Two experiments were conducted to investigate the effect of in ovo zinc （Zn） injection on the embryonic development, tissue Zn contents, antioxidation and related gene expressions of fertilized eggs of Arbor Acres broiler breeders. Experiment 1 was conducted to determine an optimal embryonic age for early in ovo injection. A total of 720 fertilized eggs with similar weights were randomly allotted to 4 treatments with 6 replicates per treatment and 30 eggs per replicate in a completely randomized design. The eggs were injected with 0.1 mL sterilized water at 3, 6 and 9 embryonic days of incubation （E3, E6 and E9） or non-injection （the control）, respectively. The results from experiment 1 showed that E3 and E6 injections increased （P〈0.05） the embryonic mortalities, and decreased （P〈0.05） hatchabilities compared to the non-injected control, but no differences （P〉0.05） between E9 injection and the non-injected control were observed in either embryonic mortality or hatchability. The findings suggest that the E9 is the optimal embryonic age for early in ovo injection. In experiment 2, a total of 672 fertilized eggs with similar weights were randomly allocated to 7 treatments with 6 replicates per treatment and 16 eggs per replicate in a completely randomized design. The eggs were injected with 0 （the negative control）, 50, 100, 150, 200, or 250 μg Zn/egg as reagent grade ZnSO4.7H20 in a 0.1-mL solution, or non-injection （the positive control）, respectively at E9-10. The results from the experiment 2 demonstrated that no differences （P〉0.05） among 50 and 100 μg Zn/egg groups and the negative control were observed in the embryonic mortality and hatchability, however, the injection of 200 μg Zn/egg increased （P〈0.05） the embryonic mortality, and injections of 150 and 200 μg Zn/egg decreased （P〈0.05） hatchabilities compared with the controls. The embryonic tibia Zn contents at E20 were increased （P〈0.05） by injections of 150, 200 and 250μg Zn/egg. Zinc injection did not affect （P〉0.05） malonaldehyde （MDA） contents, copper- and Zn- containing superoxide dismutase （CuZnSOD） activities and mRNA expression levels in the liver and heart of chick embryos at E15 and E20. Compared with the negative control, injections of 50, 150 and 200 μg Zn/egg up-regulated （P〈0.05） themetallothionein （MT） mRNA expression levels in the embryonic liver at E20. These results indicated that in ovo Zn injections increased Zn contents in the embryonic tibia and MT mRNA expression levels in the embryonic liver at E20, however, injections of 150-200 μg Zn/egg were harmful to the embryonic development.
基金funded from the European Union’s Seventh Framework Programme for research,technological development and demonstration under grant agreement n°311,794co-financed from funds for science of the Polish Ministry of Science and Education allocated to an international project ECO FCE in the years 2013–2017
文摘Background： Application the innovative method which is in ovo technology provides a means of modulating the immune system at early embryonic stages. The aim of study was to determine influence of the in ovo stimulation, on d12 of incubation, with synbiotics（synbiotic 1-L. salivarius IBB3154 ＋ Bi^2tos, Clasado Ltd. and the synbiotic 2-L. plantarum IBB3036 ＋ lupin RFOs） on the microstructure of duodenum, jejunum and ileum in the 1^（st） and 42^（nd）day of rearing.Results： On the 1^（st） day of chickens life, in the duodenum of both experimental groups（SYN1 and SYN2）, a significantly higher and wider intestinal vil i as wel as a significantly larger absorbent surface of these villi were found in comparison with the Control group（P ≤ 0.01）. On the 42^（nd）day of rearing the beneficial effect of synbiotic 1 was reflected by the numerical y higher vil i（no statistical differences） with a larger surface（P ≤ 0.01） in the duodenum in the SYN1 group compare to the Control group. In the jejunum on the 1^（st） day of life, in the SYN1 group, significantly higher vil i than in the Control group, with a simultaneous decrease in the depth of crypts（P ≤ 0.01）, and also the largest width of vil i and their absorbent area（P ≤ 0.01） in comparison to the other groups were found. On the 42^（nd）day of life, in the jejunum, an increase in the height of the vil i whilst reducing the crypt depth in the SYN2 group was found（P ≤ 0.01）. In turn, in the SYN1 group, there were significantly more neutral goblet cel s observed compared with the control group（P ≤ 0.05）. In the ileum of 1-day-old chickens, the widest vil i（P ≤ 0.05） and the deepest crypts（P ≤ 0.01） were found in the SYN2 group. In the same group, there was also the least amount of neutral goblet cel s in comparison to the other groups（P ≤ 0.05）.Conclusions： We observed that synbiotic 1 and 2 beneficial y affected the examined characteristics on the 1^（st） and 42^（nd） day of life. The obtained results al ow us to conclude that the use of synbiotics significantly affect gut structure which should contribute to improvement in nutrient absorption by the gut.
基金supported by the NSFC-Guangdong Joint Fund (U0931004)the Sino-German Cooperation in Agriculture (Project No. 28/04-05CHN7,2010-2011)+4 种基金the Special Fund for Agro-scientific Research in the Public Interest (201003011)the National "948" project (2011-S11)the Natural Science Foundation of Jiangsu Province (BK2010296)the Doctor Foundation of the Institute of Poultry Science of Jiangsu Province (JQ201001)the Priority Academic Program Development of Jiangsu Higher Education Institutions
文摘Background： A leptin-like immunoreactive substance has been found in chicken eggs and has been implicated in serving as a maternal signal to program offspring growth and metabolism. In the present study, we investigated the effects of in ovo leptin administration on hatch weight, serum and hepatic concentrations of metabolites and hormones, as well as on the expression of genes involved in hepatic lipid metabolism and the predicted microRNAs （miRNAs） targeting the affected genes. To this end we injected fertile eggs with either 0.5 μg of recombinant murine leptin or vehicle （PBS） before incubation.Results： Prenatally leptin-exposed chicks showed lower hatch weight, but higher liver weight relative to the body weight, compared to the control group. In ovo leptin treatment increased the hepatic content and serum concentration of leptin in newly hatched chickens. The hepatic contents of triglycerides （TG） and total cholesterol （Tch） were decreased, whereas the serum levels of TG, Tch and apolipoprotein B （ApoB） were increased. The hepatic mRNA expression of sterol regulator element binding protein 1 （SREBP-1c）, SREBP-2, hydroxy-3-methylglutaryl coenzyme A reductase （HMGCR） and cholesterol 7a-hydroxylase 1 （CYP7A1） was significantly up-regulated, as was the protein content of both SREBP-1 c and SREBP-2 in hepatic nuclear extracts of leptin-treated chickens. Moreover, out of 12 miRNAs targeting SREBP-1c and/or HMGCR, five were significantly up-regulated in liver of leptin-treated chicks, including gga-miR-200b and gga-miR-429, which target both SREBP-1c and HMGCR.Conclusions： These results suggest that leptin in ovo decreases hatch weight, and modifies hepatic leptin secretion and lipid metabolism in newly hatched broiler chickens, possibly via microRNA-mediated gene regulation.
基金the Foundation for Food and Agricultural Research[EggTech-0000000028]the Research Foundation-Flanders[SB project 1SC7219N and SB project 1S54823N].
文摘Numerous researchers and institutions have been developing in ovo sexing technologies to improve animal welfare by identifying male embryos in an early embryonic stage and disposing of them before pain perception.This review gives a complete overview of the technological approaches reported in papers and patents by performing a thorough search using Web of Science and Patstat/Espacenet databases for papers and patents,respectively.Based on a total of 49 papers and 115 patent families reported until May 2023 worldwide,11 technology categories were defined:6 non-optical and 5 optical techniques.Every category was described for its characteristics while assessing its potential for application.Next,the dynamics of the publications of in ovo sexing techniques in both paper and patent fields were described through growth curves,and the interest or actual status was visualized using the number of paper citations and the actual legal status of the patents.When comparing the reported technologies in papers to those in patents,scientific gaps were observed,as some of the patented technologies were not reported in the scientific literature,e.g.,ion mobility and mass spectrometry approaches.Generally,more diverse approaches in all categories were found in patents,although they do require more scientific evidence through papers or industrial adoption to prove their robustness.Moreover,although there is a recent trend for non-invasive techniques,invasive methods like analyzing DNA through PCR or hormones through immunosensing are still being reported(and might continue to be)in papers and patents.It was also observed that none of the technologies complies with all the industry requirements,although 5 companies already entered the market.On the one hand,more research and harmony between consumers,industry,and governments is necessary.On the other hand,close monitoring of the market performance of the currently available techniques will offer valuable insights into the potential and expectations of in ovo sexing techniques in the poultry industry.
基金funded by the National Key Research and Development Program of China(2017YFD0500500,20170502200)the National Natural Science Foundation of China(No.31972529)+1 种基金the Innovative Transforming Project of Shaanxi Agricultural Achievements in Science&Technology(NYKJ-2018-YL15)the Program for Shaanxi Science&Technology from Shaanxi Provincial Science and Technology Department(2021TD-30).
文摘Background:Many researches about in ovo feeding(IOF)of vitamin C(VC)are gradually carried out to explore physiological development in chicken,but little studies focus on VC synthesis capacity of the embryo itself,the selection of injection site and the effectiveness of IOF of VC.This study aims to explore the above problems.Results:Kidney and yolk sac were the main organs for VC synthesis and L-gulonolactone oxidase(GLO)expression was lower during pre-hatch development than that during post-hatch development.Sodium-dependent vitamin C transporter 1(SVCT1)expression was increased continuously in yolk sac from embryonic age 19(E19)to post-hatch day 1(D1)and in intestine(duodenum,jejunum and ileum)from E17 to D1.Plasma VC content was higher at D1 than that at D21 and D42.IOF of VC significantly reduced GLO expression in liver,kidney and yolk sac as well as SVCT1 expression in duodenum,jejunum and ileum,but increased the VC content in plasma,brain,kidney and liver.In addition,IOF of VC obviously reduced the embryonic morality and increased the hatchability under heat stress.Conclusions:This study suggested that IOF of VC at E11 in yolk was effective for embryonic VC supplementation.These findings provide a theoretical reference about the method of embryonic VC supplementation and effective methodology on embryonic VC nutrition in broiler chickens.
基金support by AgriFutures Australia’s Chicken Meat Program[grant number PRJ-011584]is gratefully acknowledged.
文摘Background Broiler chickens are most vulnerable immediately after hatching due to their immature immune systems,making them susceptible to infectious diseases.The yolk plays an important role in early immune defence by showing relevant antioxidant and passive immunity capabilities during broiler embryonic development.The immunomodulatory effects of phytogenic compound carvacrol have been widely reported.After in ovo delivery in the amniotic fluid during embryonic development carvacrol is known to migrate to the yolk sac.However,it is unknown whether carvacrol in the yolk could enhance defence responsiveness in the yolk sac.Therefore,the aim of this study was to improve early immune function in chicken embryos,and it was hypothesized that in ovo delivery of carvacrol would result in immunomodulatory effects in the yolk sac,potentially improving post-hatch resilience.Methods On embryonic day(E)17.5,either a saline(control)or carvacrol solution was injected into the amniotic fluid.Yolk sac tissue samples were collected at E19.5,and transcriptomic analyses using RNA sequencing were performed,following functional enrichment analyses comparing the control(saline)and carvacrol-injected groups.Results The results showed that 268 genes were upregulated and 174 downregulated in the carvacrol group compared to the control(P<0.05;logFC<-0.5 or log FC>0.5).Functional analyses of these differentially expressed genes,using KEGG,REACTOME,and Gene Ontology databases,showed enrichment of several immune-related pathways.This included the pathways‘Antimicrobial peptides’(P=0.001)and‘Chemoattractant activity’(P=0.004),amongst others.Moreover,the‘NOD-like receptor signaling’pathway was enriched(P=0.002).Antimicrobial peptides are part of the innate immune defence and are amongst the molecules produced after the nucleotide oligomeriza-tion domain(NOD)-like receptor pathway activation.While these responses may be associated with an inflammatory reaction to an exogenous threat,they could also indicate that in ovo delivery of carvacrol could prepare the newly hatched chick against bacterial pathogens by potentially promoting antimicrobial peptide production through acti-vation of NOD-like receptor signaling in the yolk sac.Conclusion In conclusion,these findings suggest that in ovo delivery of carvacrol has the potential to enhance anti-pathogenic and pro-inflammatory responses in the yolk sac via upregulation of antimicrobial peptides,and NOD-like receptor pathways.
文摘BACKGROUND Cardiovascular diseases are the first cause of death in the world.Ischemic heart disease is the main cause of heart failure.New approaches are continuously sou-ght to identify better therapeutic success.Thereby,current research has been drawn to identifying and completing the therapeutic profile of natural sources.Galium species are representatives exhibiting diuretic and antibacterial potential in living organisms and can treat burns,wounds,and skin diseases.Moreover,it was also observed that these plants manifest cardioprotective effects as well as having antihemolytic,antioxidant,antibacterial,anti-inflammatory,immunomod-ulatory,and antiproliferative potential.In ischemic heart disease,Galium verum(G.verum)extract manifested preservative properties in terms of contractility,systolic and diastolic function maintenance,and reduced damage to the heart after ischemia.In addition,G.verum extract upregulated the activity of antioxidant enzymes alleviating the production of pro-oxidants.AIM To test the ethanolic extract of G.verum on the H9C2(2-1)cell line by evaluating the in vitro biosafety profile and in ovo irritative potential.METHODS Cells were tested in vitro for viability(using the MTT test),cellular morphology,cell number,confluence,nuclear morphology(by immunofluorescence staining of cell nuclei and F-actin assay)and in ovo by the hen’s egg chorioal-lantoic membrane(CAM)test and CAM anti-irritant methods to study the irritation potential on the CAM.RESULTS The extract demonstrated a dose-dependent stimulatory activity.The viability increased to 170%for the dose of 55µg/mL and decreased to 135%at 200µg/mL.The results of cell number,confluence,and morphological analysis did not present significant changes compared with control untreated cells.The immunofluorescence assay showed insignificant apoptotic potential,and the hen’s egg CAM test revealed that the extract was in the weak to mode-rately irritating category with an irritation score of 5.3.When applying the sample to the CAM,only slight coagu-lation was observed(128 s).The anti-irritant test revealed the protective potential of the extract in the vascular plexus.CONCLUSION The ethanolic extract of G.verum manifests a stimulating effect on cardiomyocytes,enhancing cell viability,and maintaining a normal elongated shape,cell number,and confluence,without significant signs of apoptosis and with a weak irritative effect in ovo.In addition,the extract demonstrated a protective effect against hemorrhage,lysis,and coagulation of blood vessels induced by sodium dodecyl sulfate on the CAM.
文摘Background Hatch weight(HW)affects broiler growth and low HW(LHW)often leads to suboptimal performance.Sodium butyrate(SB)has been shown to promote growth through enhanced intestinal health.This study investi-gated how broilers with different HW responded to in ovo SB injection and whether SB could enhance gut health and performance in LHW chicks.Ross 308 broiler eggs were injected on incubation d 12 with physiological saline(control)or SB at 0.1%(SB1),0.3%(SB3),or 0.5%(SB5).Post-hatch,male chicks from each treatment were categorized as high HW(HHW)or LHW and assigned to 8 groups in a 4×2 factorial design.Production parameters were recorded periodically.Intestinal weight,length,and gene expression related to gut barrier function and immune response were examined on d 14 and 42.Cecal microbiota dynamics and predicted functionality were analyzed using 16S rRNA gene sequencing.Results SB treatments did not affect hatchability.HHW-control group exhibited consistently better weight gain and FCR than LHW-control group.SB dose-dependently influenced performance and gut health in both HW catego-ries,with greater effects in LHW broilers at 0.3%.LHW-SB3 group attained highest body weight on d 42,exceeding controls but not significantly differing from HHW-SB3 group.LHW-SB3 group showed upregulation of gut-barrier genes CLDN1 in ileum,TJP1 in jejunum and anti-inflammatory cytokine IL-10 in both jejunum and ileum on d 14.Addi-tionally,LHW-SB3 group upregulated mucin-producing MUC6 gene in ileum,while HHW-SB5 group increased pro-inflammatory IL-12p40 cytokine in caecum on d 42.LHW-SB3 group demonstrated shorter relative intestinal lengths,while HHW-SB5 had longer lengths.HHW-control group had higher bacterial diversity and growth-promoting bacte-ria while LHW-control group harbored the potential pathogen Helicobacter.SB reshaped gut microbiota biodiversity,composition,and predicted metabolic pathways in both HW categories.The LHW-SB3 group exhibited highest alpha diversity on d 14 and most beneficial bacteria at all timepoints.HHW-SB5 group presented increased pathogenic Escherichia-Shigella and Campylobacter on d 42.Conclusions HW significantly affects subsequent performance and SB has differential effects based on HW.LHW chicks benefited more from 0.3%SB,showing improvements in growth,intestinal development,health,and gut microbiota characteristics.
基金supported by the Open Project of Jiangsu Key Laboratory of Animal genetic Breeding and Molecular Design(No.AGBMD202202)the Jiangsu Agricultural Science and Technology Innovation Fund(Nos.[CX(21)3131]and[CX(20)3010])+2 种基金the“JBGS”Project of Seed Industry Revitalization in Jiangsu Province(Nos.[JBGS[2021]027 and JBGS[2021]105])the Science and Education Integration Project of Yangzhou University(No.[KJRH202115])Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)。
文摘Selenium is an essential trace element for humans and animals.As the active center of selenoproteins,the addition of selenium is beneficial to enhance the antioxidant ability.However,the high cost limits the application of organic Se in agriculture animal production.Selenized glucose(SeGlu)is a newly invented organoselenium material with good stability,low toxicity and low cost.This assay found that SeGlu was able to increase selenium deposition in liver of newborn broilers,and enhance the antioxidant capacity of liver by elevating the activities of antioxidant enzymes such as total superoxide dismutase and glutathione peroxidase.This paper as the first example clarifying the mechanism of SeGlu to enhance the antioxidant ability of chicks,shows that SeGlu can be used as an organic selenium enrichment additive for early nutrition of poultry.As a cross-discipline study involving chemistry,biology and agriculture animal science,the work may be beneficial for studies in related fields and prompt the development of the selenium science.
基金supported by the National Natural Science Foundation of China(31972585)the Technical System of Poultry Industry of Guangdong Province,China(2021KJ128)。
文摘The intestinal health of chick embryos is vital for their life-long growth,and exogenous nutrition intervention may provide sufficient nutrition for embryonic development.In the present study,we investigated the effect of in ovo injection of L-methionine(L-Met)on the intestinal structure and barrier function of chick embryos.There were 4 groups of treatments:the control(CON)group injected with phosphate-buffered saline(PBS)and the other 3 groups injected with 5,10,and 20 mg L-Met/egg,respectively.The injection was performed on embryonic day 9(E9),and intestinal samples were collected on the day of hatching for analysis.The results showed that,compared with the CON group,the groups administered an in ovo injection of L-Met increased relative weights of the duodenum,jejunum,and ileum(P<0.05).Hematoxylin and eosin(H&E)staining showed that the groups injected with 5,10,and 20 mg L-Met significantly increased villus height and crypt depth(P<0.05).Moreover,in ovo injection of 10 mg L-Met also increased the transepithelial electrical resistance(TEER)of the jejunum(P<0.05).Injection with 10 and 20 mg L-Met increased the expression of the tight junction proteins(ZO-1 and claudin-1)and the fluorescence signal intensity of Ki67 and villin proteins(P<0.05).Further,the protein expression of phospho-Janus kinase 2(p-JAK2)and phospho-signal transducer and activator of transcription 3(p-STAT3)was significantly increased by 10 or 20 mg L-Met injection(P<0.05).In conclusion,the injection of L-Met,especially at a dose of 10 mg,showed beneficial effects on the intestinal integrity of chick embryos due to the activation of the JAK2/STAT3 signaling pathway.Our results may provide new insights for regulating the intestinal development of embryonic chicks and the rapid growth of chicks after hatching.
基金supported in part by the Brain Pool program funded by the Ministry of Science and ICT through the National Research Foundation of Korea(2019H1D3A1A01071142)by the Forest Science and Technology R&D Program(2020193C10-2022-BA01)provided by Korea Forest Service(Korea Forestry Promo-tion Institute).
文摘Healthy chickens are necessary to meet the ever-increasing demand for poultry meat.Birds are subjected to numerous stressful conditions under commercial rearing systems,including variations in the envi-ronmental temperature.However,it is difficult to counter the effects of global warming on the livestock industry.High environmental temperature is a stressful condition that has detrimental effects on growth and production performance,resulting in decreased feed intake,retarded growth,compromised gut health,enhanced oxidative stress,and altered immune responses.Traditional approaches include nutritional modification and housing management to mitigate the harmful effects of hot environments.Currently,broiler chickens are more susceptible to heat stress(HS)than layer chickens because of their high muscle mass and metabolic rate.In this review,we explored the possibility of in ovo manipulation to combat HS in broiler chickens.Given their short lifespan from hatching to market age,embryonic life is thought to be one of the critical periods for achieving these objectives.Chicken embryos can be modulated through either temperature treatment or nourishment to improve thermal tolerance during the rearing phase.We first provided a brief overview of the harmful effects of HS on poultry.An in-depth evaluation was then presented for in ovo feeding and thermal manipulation as emerging strategies to combat the negative effects of HS.Finally,we evaluated a combination of the two methods using the available data.Taken together,these investigations suggest that embryonic manipulation has the po-tential to confer heat resistance in chickens.
基金National Natural Science Foundation of China （#90208011, #30300174, #30070856 , #30421005） National Key Basic Research and Development Program of China （#2002CB713802 , #2005CB522700） Shanghai Key Project of Basic Science Research （#04DZ14005 , #04DZ05608）.
文摘Members of the basic helix-loop-helix （bHLH） gene family play important roles in vertebrate neurogenesis. In this study, confocal microscopy-based fluorescence resonance energy transfer （FRET） is used to monitor bHLH protein-protein interactions under various physiological conditions. Tissue-specific bHLH activators, NeuroD 1, Mash 1, Neurogenin 1 （Ngn 1）, Neurogenin2 （Ngn2）, and ubiquitous expressed E47 protein are tagged with enhanced yellow fluorescence protein （EYFP） and enhanced cyan fluorescence protein （ECFP）, respectively. The subcellular localization and mobility ofbHLH fusion proteins are examined in HEK293 cells. By transient transfection and in ovo electroporation, four pairs of tissue-specific bHLH activators and E47 protein are over-expressed in HEK293 cells and developing chick embryo neural tube. With the acceptor photobleaching method, FRET could be detected between these bHLH protein pairs in the nuclei of transfected cells and developing neural tubes. Mashl/E47 and Ngn2/E47 FRET pairs show higher FRET efficiencies in the medial and the lateral half of chick embryo neural tube, respectively. It suggests that these bHLH protein pairs formed functional DNA-protein complexes with regulatory elements of their downstream target genes in the specific regions. This work will help one understand the behaviours of bHLH factors in vivo.
基金awarded a Stipendium Hungaricum Scholarship for Ph.D.studiesthe support of the OTKA grant(K139021)
文摘Methionine(Met)is an essential and first limiting amino acid in the poultry diet that plays a significant role in chicken embryonic development and growth.The present study examined the effect of in ovo injection of DL-Met and L-Met sources and genotypes on chicken embryonic-intestinal development and health.Fertilized eggs of the two genotypes,TETRA-SL layer hybrid(TSL)—commercial layer hybrid and Hungarian Partridge colored hen breed(HPC)—a native genotype,were randomly distributed into four treatments for each genotype.The treatment groups include the following:1)control non-injected eggs(NoIn);2)saline-injected(SaIn);3)DL-Met injected(DLM);and 4)L-Met injected(LM).The in ovo injection was carried out on 17.5 d of embryonic development;after hatching,eight chicks per group were sacrificed,and the jejunum was extracted for analysis.The results showed that both DLM and LM groups had enhanced intestinal development as evidenced by increased villus width,villus height,and villus area(P<0.05)compared to the control.The DLM group had significantly reduced crypt depth,glutathione(GSH)content,glutathione S-transferase 3 alpha(GST3),occludin(OCLN)gene expression and increased villus height to crypt depth ratio in the TSL genotype than the LM group(P<0.05).The HPC genotype has overexpressed insulin-like growth factor 1(IGF1)gene,tricellulin(MD2),occludin(OCLN),superoxide dismutase 1(SOD1),and GST3 genes than the TSL genotype(P<0.05).In conclusion,these findings showed that in ovo injection of Met enhanced intestinal development,and function,with genotypes responding differently under normal conditions.Genotypes also influenced the expression of intestinal antioxidants,tight junction,and growth-related genes.
