Responses of growth performance,antioxidant function,small intestinal morphology and mRNA expression of jejunal tight junction protein to dietary iron in yellow-feathered broilers

This study aimed to investigate the dose-effect of iron on growth performance,antioxidant function.intestinal morphology,and mRNA expression of jejunal tight junction protein in 1-to21-d-old yellow-feathered broilers.A total of 7201-d-old yellow-feathered maleb roilers were allocated to 9 treatments with 8 replicate cages of 10 birds per cage.The dietary treatments were consisted of a basal diet(contained 79.6 mg Fe kg^(-1))supplemented with 0,20,40,60,80,160,320,640,and 1,280 mg Fe kg^(-1)in the form of FeSO_(4)·7H_(2)O.Compared with the birds in the control group,birds supplemented with 20mg Fe kg^(-1)had higher average daily gain(ADG)(P<0.0001).Adding 640 and 1,280 mg Fe kg^(-1)significantly decreased ADG(P<0.0001)and average daily feed intake(ADFI)(P<0.0001)compared with supplementation of 20mg Fe kg^(-1).Malondialdehyde(MDA)concentration in plasma and duodenum increased linearly(P<0.0001),but MDA concentration in liver and jejunum increased linearly(P<0.05)or quadratically(P<0.05)with increased dietary Fe concentration.The villus height(VH)in duodenum and jejunum,and the ratio of villus height to crypt depth(V/C)in duodenum decreased linearly(P?0.05)as dietary Feincreased.As dietary Fe increased,the jejunal relative mRNA abundance of claudin-1 decreased linearly(P=0.001),but the jejunal relative mRNA abundance of zona occludens-1(ZO-1)and occludin decreased linearly(P?0.05)or quadratically(P?0.05).Compared with the supplementation of 20 mg Fe kg^(-1),the supplementation of640 mg Fe kg^(-1)or higher increased(P?0.05)MDA concentrations in plasma,duodenum,and jejunum,decreased VH in the duodenum and jejunum,and the addition of 1,280 mg Fe kg^(-1)reduced(P?0.05)the jejunal tight junction protein(claudin-1,ZO-1,occludin)mRNA abundance.In summary,640 mg of supplemental Fe kg^(-1)or greater was associated with decreased growth performance,increased oxidative stress,disrupted intestinal morphology,and reduced mRNA expression of jejunal tight junction protein.

The chemical characteristics of different sodium iron ethylenediaminetetraacetate sources and their relative bioavailabilities for broilers fed with a conventional corn‑soybean meal diet

Background Our previous studies demonstrated that divalent organic iron(Fe)proteinate sources with higher complexation or chelation strengths as expressed by the greater quotient of formation(Qf)values displayed higher Fe bioavailabilities for broilers.Sodium iron ethylenediaminetetraacetate(NaFeEDTA)is a trivalent organic Fe source with the strongest chelating ligand EDTA.However,the bioavailability of Fe when administered as NaFeEDTA in broilers and other agricultural animals remains untested.Herein,the chemical characteristics of 12 NaFeEDTA products were determined.Of these,one feed grade NaFeEDTA(Qf=2.07×10^(8)),one food grade NaFeEDTA(Qf=3.31×10^(8)),and one Fe proteinate with an extremely strong chelation strength(Fe-Prot ES,Qf value=8,590)were selected.Their bioavailabilities relative to Fe sulfate(FeSO_(4)·7H_(2)O)for broilers fed with a conventional corn-soybean meal diet were evaluated during d 1 to 21 by investigating the effects of the above Fe sources and added Fe levels on the growth performance,hematological indices,Fe contents,activities and gene expressions of Fe-containing enzymes in various tissues of broilers.Results NaFeEDTA sources varied greatly in their chemical characteristics.Plasma Fe concentration(PI),transferrin saturation(TS),liver Fe content,succinate dehydrogenase(SDH)activities in liver,heart,and kidney,catalase(CAT)activity in liver,and SDH mRNA expressions in liver and kidney increased linearly(P<0.05)with increasing levels of Fe supplementation.However,differences among Fe sources were detected(P<0.05)only for PI,liver Fe content,CAT activity in liver,SDH activities in heart and kidney,and SDH mRNA expressions in liver and kidney.Based on slope ratios from multiple linear regressions of the above indices on daily dietary analyzed Fe intake,the average bioavailabilities of Fe-Prot ES,feed grade NaFeEDTA,and food grade NaFeEDTA relative to the inorganic FeSO_(4)·7H_(2)O(100%)for broilers were 139%,155%,and 166%,respectively.Conclusions The bioavailabilities of organic Fe sources relative to FeSO_(4)·7H_(2)O were closely related to their Qf values,and NaFeEDTA sources with higher Qf values showed higher Fe bioavailabilities for broilers fed with a conventional corn-soybean meal diet.

Effects of Enteromorpha prolifera polysaccharides on growth performance,intestinal barrier function and cecal microbiota in yellow-feathered broilers under heat stress

Background Global warming leading to heat stress(HS)is becoming a major challenge for broiler production.This study aimed to explore the protective effects of seaweed(Enteromorpha prolifera)polysaccharides(EPS)on the intestinal barrier function,microbial ecology,and performance of broilers under HS.A total of 144 yellow-feathered broilers(male,56 days old)with 682.59±7.38 g were randomly assigned to 3 groups:1)TN(thermal neutral zone,23.6±1.8℃),2)HS(heat stress,33.2±1.5℃ for 10 h/d),and 3)HSE(HS+0.1%EPS).Each group contained 6 replicates with 8 broilers per replicate.The study was conducted for 4 weeks;feed intake and body weights were measured at the end of weeks 2 and 4.At the end of the feeding trial,small intestine samples were collected for histomorphology,antioxidant,secretory immunoglobulin A(s Ig A)content,apoptosis,gene and protein expression analysis;cecal contents were also collected for microbiota analysis based on 16S r DNA sequencing.Results Dietary EPS promoted the average daily gain(ADG)of broilers during 3–4 weeks of HS(P<0.05).At the end of HS on broilers,the activity of total superoxide dismutase(T-SOD),glutathione S-transferase(GST),and the content of s Ig A in jejunum were improved by EPS supplementation(P<0.05).Besides,dietary EPS reduced the epithelial cell apoptosis of jejunum and ileum in heat-stressed broilers(P<0.05).Addition of EPS in HS group broilers'diet upregulated the relative m RNA expression of Occludin,ZO-1,γ-GCLc and IL-10 of the jejunum(P<0.05),whereas downregulated the relative m RNA expression of NF-κB p65,TNF-αand IL-1βof the jejunum(P<0.05).Dietary EPS increased the protein expression of Occludin and ZO-1,whereas it reduced the protein expression of NF-κB p65 and MLCK(P<0.01)and tended to decrease the protein expression of TNF-α(P=0.094)in heat-stressed broilers.Furthermore,the proportions of Bacteroides and Oscillospira among the three groups were positively associated with jejunal apoptosis and pro-inflammatory cytokine expression(P<0.05)and negatively correlated with jejunal Occludin level(P<0.05).However,the proportions of Lactobacillus,Barnesiella,Subdoligranulum,Megasphaera,Collinsella,and Blautia among the three groups were positively related to ADG(P<0.05).Conclusions EPS can be used as a feed additive in yellow-feathered broilers.It effectively improves growth performance and alleviates HS-induced intestinal injury by relieving inflammatory damage and improving the tight junction proteins expression.These beneficial effects may be related to inhibiting NF-κB/MLCK signaling pathway activation and regulation of cecal microbiota.

Transcriptome analysis reveals steroid hormones biosynthesis pathway involved in abdominal fat deposition in broilers

Excessive abdominal fat deposition reduces the feed efficiency and increase the cost of production in broilers.Therefore,it is an important task for poultry breeders to breed broilers with low abdominal fat.Abdominal fat deposition is a highly complex biological process,and its molecular basis remains elusive.In this study,we performed transcriptome analysis to compare gene expression profiles at different stages of abdominal fat deposition to identify the key genes and pathways involved in abdominal fat accumulation.We found that abdominal fat weight(AFW)increased gradually from day 35(D35)to 91(D91),and then decreased at day 119(D119).Accordingly,after detecting differentially expressed genes(DEGs)by comparing gene expression profiles at D35 vs.D63 and D35 vs.D91,and identifying gene modules associated with fat deposition by weighted gene co-expression network analysis(WGCNA),we performed intersection analysis of the detected DEGs and WGCNA gene modules and identified 394 and 435 intersecting genes,respectively.The results of the Gene Ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses showed that the steroid hormone biosynthesis and insulin signaling pathways were co-enriched in all intersecting genes,steroid hormones have been shown that regulated insulin signaling pathway,indicating the importance of the steroid hormone biosynthesis pathway in the development of broiler abdominal fat.We then identified 6 hub genes(ACTB,SOX9,RHOBTB2,PDLIM3,NEDD9,and DOCK4)related to abdominal fat deposition.Further analysis also revealed that there were direct interactions between 6 hub genes.SOX9 has been shown to bind to proteins required for steroid hormone receptor binding,and RHOBTB2 indirectly regulates the steroid hormones biosynthesis through cyclin factor,and ultimately affect fat deposition.Our results suggest that the genes RHOBTB2 and SOX9 play an important role in fat deposition in broilers,by regulating steroid hormone synthesis.These findings provide new targets and directions for further studies on the mechanisms of fat deposition in chicken.

Hypoxia-mediated programmed cell death is involved in the formation of wooden breast in broilers

Background Wooden breast(WB)myopathy is a common myopathy found in commercial broiler chickens worldwide.Histological examination has revealed that WB myopathy is accompanied by damage to the pectoralis major(PM)muscle.However,the underlying mechanisms responsible for the formation of WB in broilers have not been fully elucidated.This study aimed to investigate the potential role of hypoxia-mediated programmed cell death(PCD)in the formation of WB myopathy.Results Histological examination and biochemical analysis were performed on the PM muscle of the control(CON)and WB groups.A significantly increased thickness of the breast muscle in the top,middle,and bottom portions(P<0.01)was found along with pathological structure damage of myofibers in the WB group.The number of capillaries per fiber in PM muscle,and the levels of p O_(2) and s O_(2) in the blood,were significantly decreased(P<0.01),while the levels of p CO_(2) and TCO_(2) in the blood were significantly increased(P<0.05),suggesting hypoxic conditions in the PM muscle of the WB group.We further evaluated the PCD-related pathways including autophagy,apoptosis,and necroptosis to understand the consequence response to enhanced hypoxic conditions in the PM muscle of birds with WB.The ratio of LC3 II to LC3 I,and the autophagy-related factors HIF-1α,BNIP3,Beclin1,AMPKα,and ULK1 at the m RNA and protein levels,were all significantly upregulated(P<0.05),showing that autophagy occurred in the PM muscle of the WB group.The apoptotic index,as well as the expressions of Bax,Cytc,caspase 9,and caspase 3,were significantly increased(P<0.05),whereas Bcl-2 was significantly decreased(P<0.05)in the WB-affected PM muscle,indicating the occurrence of apoptosis mediated by the mitochondrial pathway.Additionally,the expressions of necroptosis-related factors RIP1,RIP3,and MLKL,as well as NF-κB and the pro-inflammatory cytokines TNF-α,IL-1β,and IL-6,were all significantly enhanced(P<0.05)in the WB-affected PM muscle.Conclusions The WB myopathy reduces blood supply and induces hypoxia in the PM muscle,which is closely related to the occurrence of PCD including apoptosis,autophagy,and necroptosis within myofibers,and finally leads to abnormal muscle damage and the development of WB in broilers.

Effects of methionine supplementation in a reduced protein diet on growth performance, oxidative status, intestinal health, oocyst shedding, and methionine and folate metabolism in broilers under Eimeria challenge

Background This study investigated effects of different methionine(Met)supplementation levels in a reduced protein diet on growth performance,intestinal health,and different physiological parameters in broilers under Eimeria challenge.A total of 600 fourteen-day-old Cobb500 male broilers were challenged with E.maxima,E.tenella,and E.acervulina,and randomly allocated in a 2×5 factorial arrangement.Birds received normal protein diets(20%crude protein,NCP)or reduced protein diets(17%crude protein,LCP),containing 2.8,4.4,6.0,7.6,and 9.2 g/kg of Met.Results On 6 and 9 days post inoculation(DPI),increasing Met level linearly improved the growth performance(P<0.05).Total oocyst shedding linearly increased as Met level increased(P<0.05).Duodenal villus height(VH):crypt depth(CD)in the LCP groups were higher on 6 DPI(P<0.01)while lower on 9 DPI(P<0.05)compared to the NCP groups.Jejunal CD and duodenal VH:CD changed quadratically as Met level increased(P<0.05).On 6 DPI,liver glutathione(GSH)and glutathione disulfide(GSSG)linearly increased as Met level increased(P<0.05).On 9 DPI,GSSG quadratically increased,whereas GSH:GSSG quadratically decreased as Met levels increased(P<0.05).The expression of amino acid transporters linearly decreased as Met level increased(P<0.05).The expression of zonula occludens 2 and claudin-1 linearly increased on 6 DPI whereas decreased on 9 DPI as Met level increased(P<0.05).The expressions of cytokines were lower in the LCP groups than the NCP groups(P<0.05).Interaction effects were found for the expression of IL-10 and TNFαon 6 DPI(P<0.05),where it only changed quadratically in the NCP group as Met level increased.The expression of Met and folate metabolism genes were lower in the LCP groups than the NCP groups on 9 DPI(P<0.05).The expression of these genes linearly or quadratically decreased as Met level increased(P<0.05).Conclusion These results revealed the regulatory roles of Met in different physiological parameters including oxidative status,intestinal health,and nutrient metabolism in birds fed reduced protein diet and challenged with Eimeria.

Dietary manganese supplementation inhibits abdominal fat deposition possibly by regulating gene expression and enzyme activity involved in lipid metabolism in the abdominal fat of broilers

Excessive abdominal fat deposition seriously restricts the production efficiency of broilers.Several studies found that dietary supplemental manganese(Mn)could effectively reduce the abdominal fat deposition of broilers,but the underlying mechanisms remain unclear.The present study aimed to investigate the effect of dietary supplementation with the inorganic or organic Mn on abdominal fat deposition,and enzyme activity and gene expression involved in lipid metabolism in the abdominal fat of male or female broilers.A total of 4201-d-old AA broilers(half males and half females)were randomly allotted by body weight and gender to 1 of 6 treatments with 10 replicates cages of 7 chicks per cage in a completely randomized design involving a 3(dietary Mn addition)×2(gender)factorial arrangement.Male or female broilers were fed with the Mn-unsupplemented basal diets containing 17.52 mg Mn kg^(-1)(d 1-21)and 15.62 mg Mn kg^(-1)(d 22-42)by analysis or the basal diets supplemented with 110 mg Mn kg^(-1)(d 1-21)and 80 mg Mn kg^(-1)(d 22-42)as either the Mn sulfate or the Mn proteinate with moderate chelation strength(Mn-Prot M)for 42 d.The results showed that the interaction between dietary Mn addition and gender had no impact(P>0.05)on any of the measured parameters;abdominal fat percentage of broilers was decreased(P<0.003)by Mn addition;Mn addition increased(P<0.004)adipose triglyceride lipase(ATGL)activity,while Mn-Prot M decreased(P<0.002)the fatty acid synthase(FAS)activity in the abdominal fat of broilers compared to the control;Mn addition decreased(P<0.009)diacylglycerol acyltransferase 2(DGAT2)mRNA expression level and peroxisome proliferator-activated receptor γ(PPARγ)mRNA and protein expression levels,but up-regulated(P<0.05)the ATGL mRNA and protein expression levels in the abdominal fat of broilers.It was concluded that dietary supplementation with Mn inhibited the abdominal fat deposition of broilers possibly via decreasing the expression of PPARγand DGAT2 as well as increasing the expression and activity of ATGL in the abdominal fat of broilers,and Mn-Prot M was more effective in inhibiting the FAS acitivity.

Transcriptomic and epigenomic landscapes of muscle growth during the postnatal period of broilers

Background Broilers stand out as one of the fastest-growing livestock globally,making a substantial contribution to animal meat production.However,the molecular and epigenetic mechanisms underlying the rapid growth and development of broiler chickens are still unclear.This study aims to explore muscle development patterns and regulatory networks during the postnatal rapid growth phase of fast-growing broilers.We measured the growth performance of Cornish(CC)and White Plymouth Rock(RR)over a 42-d period.Pectoral muscle samples from both CC and RR were randomly collected at day 21 after hatching(D21)and D42 for RNA-seq and ATAC-seq library construction.Results The consistent increase in body weight and pectoral muscle weight across both breeds was observed as they matured,with CC outpacing RR in terms of weight at each stage of development.Differential expression analysis identified 398 and 1,129 genes in the two dimensions of breeds and ages,respectively.A total of 75,149 ATAC-seq peaks were annotated in promoter,exon,intron and intergenic regions,with a higher number of peaks in the promoter and intronic regions.The age-biased genes and breed-biased genes of RNA-seq were combined with the ATAC-seq data for subsequent analysis.The results spotlighted the upregulation of ACTC1 and FDPS at D21,which were primarily associated with muscle structure development by gene cluster enrichment.Additionally,a noteworthy upregulation of MUSTN1,FOS and TGFB3 was spotted in broiler chickens at D42,which were involved in cell differentiation and muscle regeneration after injury,suggesting a regulatory role of muscle growth and repair.Conclusions This work provided a regulatory network of postnatal broiler chickens and revealed ACTC1 and MUSTN1 as the key responsible for muscle development and regeneration.Our findings highlight that rapid growth in broiler chickens triggers ongoing muscle damage and subsequent regeneration.These findings provide a foundation for future research to investigate the functional aspects of muscle development.

Gut microbiota intervention attenuates thermogenesis in broilers exposed to high temperature through modulation of the hypothalamic 5‑HT pathway

Background Broilers have a robust metabolism and high body temperature,which make them less tolerant to hightemperature(HT)environments and more susceptible to challenges from elevated temperatures.Gut microbes,functioning as symbionts within the host,possess the capacity to significantly regulate the physiological functions and environmental adaptability of the host.This study aims to investigate the effects of gut microbial intervention on the body temperature and thermogenesis of broilers at different ambient temperatures,as well as the underlying mechanism involving the"gut-brain"axis.Methods Broilers were subjected to gut microbiota interference with or without antibiotics(control or ABX)starting at 1 day of age.At 21 day of age,they were divided into 4 groups and exposed to different environments for 7 d:The control and ABX groups at room temperature(RT,24±1℃,60%relative humidity(RH),24 h/d)and the control-HT and ABX-HT groups at high temperature(HT,32±1℃,60%RH,24 h/d).Results The results demonstrated that the antibiotic-induced gut microbiota intervention increased body weight and improved feed conversion in broiler chickens(P<0.05).Under HT conditions,the microbiota intervention reduced the rectal temperature of broiler chickens(P<0.05),inhibited the expression of avUCP and thermogenesisrelated genes in breast muscle and liver(P<0.05),and thus decreased thermogenesis capacity.Furthermore,the gut microbiota intervention blunted the hypothalamic‒pituitary‒adrenal axis and hypothalamic–pituitary–thyroid axis activation induced by HT conditions.By analyzing the cecal microbiota composition of control and ABX chickens maintained under HT conditions,we found that Alistipes was enriched in control chickens.In contrast,antibioticinduced gut microbiota intervention resulted in a decrease in the relative abundance of Alistipes(P<0.05).Moreover,this difference was accompanied by increased hypothalamic 5-hydroxytryptamine(5-HT)content and TPH2 expression(P<0.05).Conclusions These findings underscore the critical role of the gut microbiota in regulating broiler thermogenesis via the gut-brain axis and suggest that the hypothalamic 5-HT pathway may be a potential mechanism by which the gut microbiota affects thermoregulation in broilers.

Correlation between Intestinal Health and Coccidiosis Prevalence in Broilers during Different Seasons of the Year in Brazil from 2012 to 2018

Coccidiosis is a disease caused by intracellular protozoan parasites, specifically belonging to the genus Eimeria. These parasites target the gastrointestinal tract in different types of hosts, causing sores in the intestinal lining. The presence of these lesions reduces the animal’s ability to digest and absorb nutrients, significantly impacting their overall performance. The current study aimed to explore the potential correlation between seasonal variations and the incidence of Eimeria spp-induced lesions in broiler chickens’ gastrointestinal tracts in Brazil from 2012 to 2018. A total of 8,607 broiler chickens, aged 14 to 42 days, were sampled from 103 poultry integrated companies in Brazil to conduct intestinal health examinations. The sampling process involved selecting 3 to 6 chickens from each poultry house for examination. The assessment included various abnormalities such as shedding of intestinal cells and excessive fluid and mucus presence, thickening and tension of the intestines, food movement, roughened mucosal surface resembling a Turkish towel in the small intestine, tissue death, duodenal inflammation, intestinal inflammation, gizzard erosion, presence of worms and bedding material, and ingestion of mealworms. During the seasons, winter exhibited the highest average occurrence of Eimeria maxima microorganisms at 52.83%, with E. acervulina following closely at 26.42% in second place. In spring, E. maxima had an occurrence of 11.31%, while in fall, E. tenella had the lowest occurrence at 6.74%. When analyzing the seasonal occurrence of Eimeria, it was observed that E. maxima micro was more common during winter compared to summer (P = 0.0491). However, no discernible variation was observed in the occurrence of the remaining species across different seasons. Research findings suggest that subclinical coccidiosis is most prevalent during the winter season in Brazil. Likewise, clinical disease caused by E. acervulina is also prevalent during this time. In contrast, E. maxima is more likely to cause clinical disease in the spring, whereas E. tenella is more commonly associated with clinical disease in the fall. Lesions induced by Eimeria spp. are associated with factors influencing the overall health of broiler intestines. These findings allow for the utilization of seasonal metrics in disease management, thereby reducing economic losses associated with the condition.

Effects of Fresh Cupressus sempervirens Leaves Infusion on Growth Performance, Intestinal Microbiota and Haemato-Biochemical Parameters in Broilers

The accumulation of growth-promoting antibiotic residues in animal products and the resistance developed by bacteria in poultry farms has led to a search for natural compounds derived from plants. This study was designed to promote the production performance of broiler chickens using fresh Cupressus sempervirens leaves infusion. Fresh Cupressus sempervirens leaves were harvested, washed, chopped and ground to a paste using a blender and fermented for three days in a closed container at a rate of 500 g/L of water. The solution obtained was filtered and added at the rate of 2, 4, 6, 8 and 10 ml/L of drinking water. The chickens fed on the graded level of the solution were compared to a control ration without an additive and positive control group supplemented with 1 g antibiotic/kg feed. At the finisher phase and throughout the study period, water intake increased significantly (P < 0.05) with increasing levels of infusion. Feed intake decreased significantly (P < 0.05) with 2 and 4 ml of infusion/L drinking water. Live weight and weight gain were significantly (P < 0.05) higher with 6 ml/L, while feed conversion significantly (P < 0.05) decreased with the same treatment compared with the control treatment without additives (T0). Carcass characteristics were not significantly (P > 0.05) affected by the inclusion of Cupressus sempervirens infusion. Haematological parameters significantly (P < 0.05) increase independently of the rate of incorporation of the infusion into the drinking water, with the exception of RBCs, MCHT and PCT. Serum content in total protein, globulins, LDL cholesterol and triglycerides were significantly (P < 0.05) high with 8 and 10 ml Cupressus sempervirens infusion/litre drinking water as compared to all other treatments. AST, ALT, urea, creatine, albumin, total cholesterol and HDL-cholesterol were not significantly affected. The lactic acid bacteria load increased significantly (P E. coli and salmonella counts decreased significantly (P < 0.05) with infusion compared to the control without additive. In conclusion, 6 ml of Cupressus sempervirens infusion can be used as an alternative to antibiotic feed additives to promote growth performance in broilers.

Technical-Economic Performance, Carcass and Meat Quality of COBB500 Broilers Fed Rations Incorporating Mango By-Product Feeds in Ivory Coast (West Africa)

The study evaluated the growth performance and carcass quality of COBB500 broilers fed two experimental rations incorporating feed and mango fine ED10 for a batch of 83 birds, ED20 (batch of 83 birds) in comparison with a commercial control ration CCD (84 birds). For this purpose, 250 day-old chicks of this breed were purchased in Korhogo and transported to Tengréla in Ivory Coast for rearing. The results show that substituting mango provender ans and mango seed flour for maize resulted in rations with costs per kgMS of 215.35FCFA, 2016.93FCFA and 320FCFA for the ED10, ED20 and CCD rations respectively. The production costs per kg of chicken (FCPkgLW) of the CCD ration were significantly higher than those of the ED10 and ED20 (p 0.05). ADGs were significantly different and decreasing from CCD, ED20 to ED10 (p 0.05). Average weights at 42nd day for ED20 and CCD were homogeneous (p > 0.05) and significantly higher than for ED10 (p 0.05). By day 49th the average weight of ED20 had reached and exceeded that of CCD by day 42nd. PSLW, HCW and CCW were significantly different and decreasing from CCW, ED20 to ED10 (p 0.05). Average hot (HCY) and cooled (CCY) carcass yields of ED20 and CCD rations were comparable (p > 0.05) and significantly higher than those of ED10 ration (p = 0.009) and 0.003). Average drumstick and neck weights were significantly higher and lower for CCD, ED20 and ED10 (p 0.05). For all other parts of the cut, the average weights of the CCD ration were significantly higher (p 0.05) than those of the ED20 and ED10 rations, which were homogeneous (p 0.05). The incorporation of mango provender and mango seed flour in broiler rations offers the prospect of reduced production costs, less competition between man and animals over cereals, and lean meat production that is more appreciated than that of chickens fed conventional feed. As it stands, the ED20 ration can be recommended as a growth diet for broilers. It would be necessary to determine the levels of anti-nutritional substances in mango feeds and to assess the in vivo digestibility of mango ingredients and the diets incorporating them.

Dietary Epimedium extract supplementation improves intestinal functions and alters gut microbiota in broilers

Background Growth-promoting antibiotics have been banned by law in the livestock and poultry breeding industry in many countries.Various alternatives to antibiotics have been investigated for using in livestock.Epimedium(EM)is an herb rich in flavonoids that has many beneficial effects on animals.Therefore,this study was planned to explore the potential of EM as a new alternative antibiotic product in animal feed.Methods A total of 7201-day-old male broilers(Arbor Acres Plus)were randomly divided into six groups and fed basal diet(normal control;NC),basal diet supplemented with antibiotic(75 mg/kg chlortetracycline;CTC),and basal diet supplemented with 100,200,400 or 800 mg/kg EM extract for 6 weeks(EM100,EM200,EM400 and EM800 groups).The growth performance at weeks 3 and 6 was measured.Serum,intestinal tissue and feces were collected to assay for antioxidant indexes,intestinal permeability,lactic acid and short-chain fatty acids(SCFAs)profiles,microbial composition,and expression of intestinal barrier genes.Results The average daily feed intake in CTC group at 1-21 d was significantly higher than that in the NC group,and had no statistical difference with EM groups.Compared with NC group,average daily gain in CTC and EM200 groups increased significantly at 1-21 and 1-42 d.Compared with NC group,EM200 and EM400 groups had significantly decreased levels of lipopolysaccharide and D-lactic acid in serum throughout the study.The concentrations of lactic acid,acetic acid,propionic acid,butyric acid and SCFAs in feces of birds fed 200 mg/kg EM diet were significantly higher than those fed chlortetracycline.The dietary supplementation of chlortetracycline and 200 mg/kg EM significantly increased ileal expression of SOD1,Claudin-1 and ZO-1 genes.Dietary supplemented with 200 mg/kg EM increased the relative abundances of g_NK4A214_group and Lactobacillus in the jejunal,while the relative abundances of Microbacterium,Kitasatospora,Bacteroides in the jejunal and Gallibacterium in the ileum decreased.Conclusion Supplementation with 200 mg/kg EM extract improved the composition of intestinal microbiota by regulating the core bacterial genus Lactobacillus,and increased the concentration of beneficial metabolites lactic acid and SCFAs in the flora,thereby improving the antioxidant capacity and intestinal permeability,enhancing the function of tight junction proteins.These beneficial effects improved the growth performance of broilers.

Dietary organic acids ameliorate high stocking density stress-induced intestinal inflammation through the restoration of intestinal microbiota in broilers

Background:High stocking density(HSD)stress has detrimental effects on growth performance,intestinal barrier function,and intestinal microbiota in intensive animal production.Organic acids(OA)are widely used as feed addi-tives for their ability to improve growth performance and intestinal health in poultry.However,whether dietary OA can ameliorate HSD stress-induced impaired intestinal barrier in broilers remains elusive.In this study,a total of 528 one-day-old male Arbor Acres broilers were randomly allocated into 3 treatments with 12 replicates per treatment including 10 birds for normal stocking density and 17 birds for HSD.The dietary treatments were as follows:1)Normal stocking density+basal diet;2)HSD+basal diets;3)HSD+OA.Results:HSD stress can induce increased levels of serum corticosterone,lipopolysaccharides,interleukin-1β,tumor necrosis factor-α,and down-regulated mRNA expression of ZO-1,resulting in compromised growth performance of broilers(P<0.05).Dietary OA could significantly reduce levels of serum corticosterone,lipopolysaccharides,interleukin-1β,and tumor necrosis factor-α,which were accompanied by up-regulated interleukin-10,mRNA expres-sion of ZO-1,and growth performance(P<0.05).Moreover,OA could down-regulate the mRNA expression of TLR4 and MyD88 to inhibit the NF-κB signaling pathway(P<0.05).Additionally,HSD stress significantly decreased the abundance of Bacteroidetes and disturbed the balance of microbial ecosystems,whereas OA significantly increased the abundance of Bacteroidetes and restored the disordered gut microbiota by reducing competitive and exploita-tive interactions in microbial communities(P<0.05).Meanwhile,OA significantly increased the content of acetic and butyric acids,which showed significant correlations with intestinal inflammation indicators(P<0.05).Conclusions:Dietary OA ameliorated intestinal inflammation and growth performance of broilers through restor-ing the disordered gut microbial compositions and interactions induced by HSD and elevating short-chain fatty acid production to inhibit the TLR4/NF-κB signaling pathway.These findings demonstrated the critical role of intestinal microbiota in mediating the HSD-induced inflammatory responses,contributing to exploring nutritional strategies to alleviate HSD-induced stress in animals.

Effect of replacement of soybean oil by Hermetia illucens fat on performance,digestibility,cecal microbiome,liver transcriptome and liver and plasma lipidomes of broilers

Background In contrast to protein-rich insect meal,the feed potential of insect fat is generally less explored and knowledge about the suitability of insect fat as a fat source specifically in broiler diets is still limited.In view of this,the present study aimed to comprehensively investigate the effect of partial(50%) and complete replacement of soybean oil with insect fat from Hermetia illucens(HI) larvae in broiler diets on performance,fat digestibility,cecal microbiome,liver transcriptome and liver and plasma lipidomes.Thus,100 male,1-day-old Cobb 500 broilers were randomly assigned to three groups and fed three different diets with either 0(group HI-0,n cens(HI) larvae fat for 35 d.= 30),2.5%(group HI-2.5,n %(HI-5.0,n = 35) or 5.0= 35) Hermetia illuResults Body weight gain,final body weight,feed intake,and feed:gain ratio during the whole period and apparent ileal digestibility coefficient for ether extract were not different between groups.Cecal microbial diversity did not differ between groups and taxonomic analysis revealed differences in the abundance of only four low-abundance bacterial taxa among groups;the abundances of phylum Actinobacteriota,class Coriobacteriia,order Coriobacteriales and family Eggerthellaceae were lower in group HI-5.0 compared to group HI-2.5(P < 0.05).Concentrations of total and individual short-chain fatty acids in the cecal digesta were not different between the three groups.Liver transcriptomics revealed a total of 55 and 25 transcripts to be differentially expressed between groups HI-5.0 vs.HI-0 and groups HI-2.5 vs.HI-0,respectively(P < 0.05).The concentrations of most lipid classes,with the exception of phosphatidylethanolamine,phosphatidylglycerol and lysophosphatidylcholine in the liver and cholesterylester and ceramide in plasma(P < 0.05),and of the sum of all lipid classes were not different between groups.Conclusions Partial and complete replacement of soybean oil with HI larvae fat in broiler diets had no effect on growth performance and only modest,but no adverse effects on the cecal microbiome and the metabolic health of broilers.This suggests that HI larvae fat can be used as an alternative fat source in broiler diets,thereby,making broiler production more sustainable.

Supplemental N-acyl homoserine lactonase alleviates intestinal disruption and improves gut microbiota in broilers challenged by Salmonella Typhimurium

Background Salmonella Typhimurium challenge causes a huge detriment to chicken production.N-acyl homoserine lactonase(AHLase),a quorum quenching enzyme,potentially inhibits the growth and virulence of Gram-negative bacteria.However,it is unknown whether AHLase can protect chickens against S.Typhimurium challenge.This study aimed to evaluate the effects of AHLase on growth performance and intestinal health in broilers challenged by S.Typhimurium.A total of 240 one-day-old female crossbred broilers(817C)were randomly divided into 5 groups(6 replicates/group):negative control(NC),positive control(PC),and PC group supplemented with 5,10 or 20 U/g AHLase.All birds except those in NC were challenged with S.Typhimurium from 7 to 9 days of age.All parameters related to growth and intestinal health were determined on d 10 and 14.Results The reductions(P<0.05)in body weight(BW)and average daily gain(ADG)in challenged birds were alleviated by AHLase addition especially at 10 U/g.Thus,samples from NC,PC and PC plus 10 U/g AHLase group were selected for further analysis.S.Typhimurium challenge impaired(P<0.05)intestinal morphology,elevated(P<0.05)ileal inflammatory cytokines(IL-1βand IL-8)expression,and increased(P<0.05)serum diamine oxidase(DAO)activity on d 10.However,AHLase addition normalized these changes.Gut microbiota analysis on d 10 showed that AHLase reversed the reductions(P<0.05)in several beneficial bacteria(e.g.Bacilli,Bacillales and Lactobacillales),along with increases(P<0.05)in certain harmful bacteria(e.g.Proteobacteria,Gammaproteobacteria,Enterobacteriaceae and Escherichia/Shigel a)in PC group.Furthermore,AHLase-induced increased beneficial bacteria and decreased harmful bacteria were basically negatively correlated(P<0.05)with the reductions of ileal IL-1βand IL-8 expression and serum DAO activity,but positively correlated(P<0.05)with the increased BW and ADG.Functional prediction revealed that AHLase abolished S.Typhimurium-induced upregulations(P<0.05)of certain pathogenicity-related pathways such as lipopolysaccharide biosynthesis,shigellosis,bacterial invasion of epithelial cells and pathogenic Escherichia coli infection of gut microbiota.Conclusions Supplemental AHLase attenuated S.Typhimurium-induced growth retardation and intestinal disruption in broilers,which could be associated with the observed recovery of gut microbiota dysbiosis.

Supplementing the early diet of broilers with soy protein concentrate can improve intestinal development and enhance short-chain fatty acid-producing microbes and short-chain fatty acids,especially butyric acid

Background:Research on nutrition in early-life commonly focuses on the maturation of the intestine because the intestinal system is crucial for ensuring continued growth.To explore the importance of early nutrition regulation in animals,soy protein concentrate(SPC)was added to the early diet of broilers to investigate its effects on amino acid digestibility,intestinal development,especially intestinal microorganisms,and broiler metabolites.A total of 192 oneday-old Arbor Acres(AA)male broilers were randomly assigned to two experimental treatments with 8 replicates of 12 birds.The control group was fed a basal diet(control),and the treatment group was fed a basal diet supplemented with 12%SPC(SPC12)during the first 10 d(starter phase).From d 11 to 21(grower phase)and d 22 to 42(finisher phase),a basal diet was fed to both treatment groups.Results:SPC reduced the pH value and acid-binding capacity of the starter diet(P<0.05,d 10);SPC in the early diet enhanced the gizzard weight(P<0.05,d 10 and d 42)and the ileum weight(P<0.05,d 10)and decreased the weight and length of the jejunum(P<0.05,d 10)and the relative length of the duodenum and jejunum(P<0.05,d 10).At the same time,SPC enhanced villus height(P<0.05,d 10)and muscle thickness in the jejunum and ileum(P<0.05,d 10)and increased the number of goblet cells in the duodenum(P<0.05,d 10).Meanwhile,SPC increased the Chao1 index and the ACE index(P<0.05,d 10)and altered the composition of caecal microflora at d 10.SPC also increased the relative abundance of Alistipes,Anaerotruncus,Erysipelatoclostridium,Intestinimonas and Flavonifractor bacteria(P<0.05,d 10).At the same time,the concentrations of caecal butyric acid and total short-chain fatty acids(SCFAs)were also increased in the SPC12 group(P<0.05,d 10).Conclusions:In summary,the results showed that supplementing the starter diet of broilers with SPC has a significant effect on the early development of the intestine and the microflora.

Heat stress in pigs and broilers:role of gut dysbiosis in the impairment of the gut-liver axis and restoration of these effects by probiotics,prebiotics and synbiotics

Heat stress is one of the most challenging stressors for animal production due to high economic losses resulting from impaired animal’s productivity,health and welfare.Despite the fact that all farm animal species are susceptible to heat stress,birds and pigs are particularly sensitive to heat stress due to either lacking or non-functional sweat glands.Con-vincing evidence in the literature exists that gut dysbiosis,a term used to describe a perturbation of commensal gut microbiota,develops in broilers and pigs under heat stress.Owing to the protective role of commensal bacteria for the gut barrier,gut dysbiosis causes a disruption of the gut barrier leading to endotoxemia,which contributes to the typical characteristics of heat stressed broilers and growing and growing-finishing pigs,such as reduced feed intake,decreased growth and reduced lean carcass weight.A substantial number of studies have shown that feeding of probiotics,prebiotics and synbiotics is an efficacious strategy to protect broilers from heat stress-induced gut barrier disruption through altering the gut microbiota and promoting all decisive structural,biochemical,and immunologi-cal elements of the intestinal barrier.In most of the available studies in heat stressed broilers,the alterations of gut microbiota and improvements of gut barrier function induced by feeding of either probiotics,prebiotics or synbiot-ics were accompanied by an improved productivity,health and/or welfare when compared to non-supplemented broilers exposed to heat stress.These findings indicate that the restoration of gut homeostasis and function is a key target for dietary interventions aiming to provide at least partial protection of broilers from the detrimental impact of heat stress conditions.Despite the fact that the number of studies dealing with the same feeding strategy in heat stressed pigs is limited,the available few studies suggest that feeding of probiotics might also be a suitable approach to enhance productivity,health and welfare in pigs kept under heat stress conditions.

Dietary oregano aqueous extract improves growth performance and intestinal health of broilers through modulating gut microbial compositions

Background Intestinal health plays a pivotal role in broiler chicken growth.Oregano aqueous extract(OAE)effec-tively exerts anti-inflammatory and antibacterial effects.However,the protective effects of OAE on intestinal health in broilers and the underlying mechanism remain unclear.This study aimed to investigate the potential effects of OAE on growth performance,the gut microbiota and intestinal health.A total of 8401-d-old male and female broilers(Arbor Acres)were randomly allocated into 6 groups as follows:basal diet(Con),Con+antibiotics(Anti,colistin sulfate 7 g/kg,roxarsone 35 g/kg),Con+400,500,600 and 700 mg/kg OAE(OAE400,OAE500,OAE600 and OAE700).Subse-quently,fermentation in vitro together with oral administration trials were carried out to further assess the function of OAE on intestinal health of broilers.Results Dietary 700 mg/kg OAE supplementation resulted in an increase(P<0.05)in body weight and a decrease(P<0.05)in feed conversion ratio when compared with the control during d 22 to 42 of the trial.OAE addition resulted in lower(P<0.05)jejunal crypt depth and mRNA expression of IL-4 and IL-10 at d 42.In addition,dietary OAE addition increased the abundance of Firmicutes(P=0.087)and Lactobacillus(P<0.05)in the cecum,and increased(P<0.05)the content of acetic acid and butyric acid.In the in vitro fermentation test,OAE significantly increased(P<0.05)the abundance of Lactobacillus,decreased(P<0.05)the abundance of unspecified_Enterobacteriaceae,and increased the content of acetic acid(P<0.05).In the oral administration trial,higher(P<0.05)IL-4 expression was found in broilers when oral inoculation with oregano fermentation microorganisms at d 42.And SIgA content in the ileum was significantly increased(P=0.073)when giving OAE fermentation supernatant.Conclusions Dietary OAE addition could maintain intestinal health and improve growth performance through enhancing intestinal mucosal immunity and barrier function mediated by gut microbiota changes.

Stimbiotic supplementation modulated intestinal inflammatory response and improved broilers performance in an experimentally-induced necrotic enteritis infection model

Background:Two experiments were conducted to establish an optimal NE challenge model and evaluate the efficacy of stimbiotic(STB)supplementation in necrotic enteritis(NE)challenged broilers.In Exp.1,a total of 120 Arbor Acres(AA)broilers(45.0±0.21 g)were randomly assigned to 6 treatments in a 3×2 factorial arrangement.Vaccine treatments included non-challenge(0),×10 the recommended dose(×10)or×20 the recommended dose(×20)by the manufacturer.Clostridium perfringens(CP)treatments were non-challenge(No)or 3 mL of 2.2×10^(7)CFU CP challenge(Yes).In Exp.2,a total of 72 AA broilers(40.17±0.27 g)were randomly assigned to 6 treatments in a 3×2 factorial arrangement.Dietary treatments included non-additive(CON),100 mg/kg STB(STB)and 100 mg/kg STB on top of a typical commercial blend including an essential oil,probiotics,and enzyme(CB).Challenge treatments included non-NE challenge(No)and NE challenge(Yes)as established in Exp.1.Results:In Exp.1,CP and vaccine challenge decreased(P<0.05)body weight(BW),body weight gain(BWG)and feed intake(FI),and increased(P<0.05)the number of broilers with diarrhea and intestinal lesions.The oral administration of×20 recommended dose of vaccines coupled with 3 mL of 2.2×10^(7)CFU CP resulted in(P<0.01)a significantly increased incidence of wet litter and intestinal lesions.Thus,this treatment was chosen as the challenge model for the successful inducement of NE in Exp.2.In Exp.2,the NE challenge negatively affected(P<0.01)growth performance,ileal morphology,immunoglobulin contents in blood,caecal microbiota in the caecum,footpad dermatitis,intestinal lesion scores,tumour necrosis factor(TNF-α)and endotoxin in the serum compared with the non-NE challenged birds.The supplementation of STB and CB in diets enhanced(P<0.05)growth performance,intestinal microbiota,and blood profiles by stimulating ileal morphology(VH and VH:CD)and propionate production in the cecum,and there were no differences in measured variables between STB and CB supplemented birds.Conclusion:Overall,these results indicate that STB supplementation was able to reduce the inflammatory response and improve the performance of NE challenged birds,and the supplementation of STB alone was as effective as a typical commercial blend containing a number of other additives.