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Hexokinase 1 and 2 mediates glucose utilization to regulate the synthesis of kappa casein via ribosome protein subunit 6 kinase 1 in bovine mammary epithelial cells

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animal Nutrition 2024年第1期16卷 338-349页

作者： Tianyu Yang Jia Guo Han Song Osmond Datsomor Yuhang Chen Maocheng Jiang Kang Zhan Guoqi Zhao Institute of animal Culture Collection and Application College of Animal Science and Technology Yangzhou University

Glucose plays a vital part in milk protein synthesis through the mTOR signaling pathway in bovine mammary epithelial cells(BMEC). The objectives of this study were to determine how glucose affects hexokinase(HK) activity in BMEC and investigate the regulatory effect of HK in kappa casein(CSN3)synthesis via the mechanistic target of rapamycin complex 1(mTORC1) signaling pathway in BMEC. For this, HK1 and HK2 were knocked out in BMEC using the CRISPR/Cas9 system. The gene and protein expression, glucose uptake, and cell proliferation were measured. We found that glucose uptake, cell proliferation, CSN3 gene expression levels, and expression of HK1 and HK2 increased with increasing glucose concentrations. Notably, glucose uptake was significantly reduced in HK2 knockout(HK2KO)BMEC treated with 17.5 mM glucose. Moreover, under the same glucose treatment conditions, the proliferative ability and abundance of CSN3 were significantly diminished in both HK1 knockout(HK1KO)and HK2KO BMEC compared with that in wild-type BEMC. We further observed that the phosphorylation levels of ribosome protein subunit 6 kinase 1(S6K1) were reduced in HK1KO and HK2KO BMEC following treatment with 17.5 mM glucose. As expected, the levels of glucose-6-phosphate and the m RNA expression levels of glycolysis-related genes were decreased in both HK1KO and HK2KO BMEC following glucose treatment. These results indicated that the knockout of HK1 and HK2 inhibited cell proliferation and CSN3 expression in BMEC under glucose treatment, which may be associated with the inactivation of the S6K1 and inhibition of glycolysis.

关键词： Glucose Hexokinase Milk protein Mechanistic target of rapamycin complex 1 signaling pathway Bovine mammary epithelial cell Kappa casein

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Molybdenum and cadmium co-induce necroptosis through Th1/Th2 imbalance-mediated endoplasmic reticulum stress in duck ovaries

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Journal of Environmental sciences 2024年第8期142卷 92-102页

作者： Ting Cui Xueyan Dai Huiling Guo Dianyun Wang Bingyan Huang Wenjing Pu Xuesheng Chu Caiying Zhang Jiangxi Provincial Key Laboratory for animal Health Institute of Animal Population HealthCollege of Animal Science and TechnologyJiangxi Agricultural UniversityNanchang 330045China

Cadmium(Cd)and excess molybdenum(Mo)pose serious threats to animal health.Our previous study has determined that Cd and/or Mo exposure can cause ovarian damage of ducks,while the specific mechanism is still obscure.To further investigate the toxic mechanism of Cd and Mo co-exposure in the ovary,forty 8-day-old female ducks were randomly allocated into four groups for 16 weeks,and the doses of Cd and Mo in basic diet per kg were as follows:control group,Mo group(100 mg Mo),Cd group(4 mg Cd),and Mo+Cd group(100 mg Mo+4 mg Cd).Cadmium sulfate 8/3-hydrate(CdSO_(4)·8/3H_(2)O)and hexaammonium molybdate((NH_(4))_(6)Mo_(7)O_(24)·4H_(2)O)were the origins of Cd and Mo,respectively.At the 16th week of the experiment,all ovary tissues were collected for the detection of related indexes.The data indicated that Mo and/or Cd induced trace element disorders and Th1/Th2 balance to divert toward Th1 in the ovary,which activated endoplasmic reticulum(ER)stress and then provoked necroptosis through triggering RIPK1/RIPK3/MLKL signaling pathway,and eventually caused ovarian pathological injuries and necroptosis characteristics.The alterations of above indicators were most apparent in the joint group.Above all,this research illustrates that Mo and/or Cd exposure can initiate necroptosis through Th1/Th2 imbalance-modulated ER stress in duck ovaries,and Mo and Cd combined exposure aggravates ovarian injuries.This research explores the molecular mechanism of necroptosis caused by Mo and/or Cd,which reveals that ER stress attenuation may be a therapeutic target to alleviate necroptosis.

关键词： Molybdenum Cadmium Th1/Th2 imbalance Endoplasmic reticulum stress Necroptosis Ovary

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Vitamin A regulates mitochondrial biogenesis and function through p38 MAPK‑PGC‑1α signaling pathway and alters the muscle fiber composition of sheep

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Journal of animal science and Biotechnology 2024年第2期15卷 898-910页

作者： Pengkang Song Jiamin Zhao Fanqinyu Li Xiaoyi Zhao Jinxin Feng Yuan Su Bo Wang Junxing Zhao College of animal science Shanxi Agricultural UniversityTaiguShanxi 030801People’s Republic of China State Key Laboratory of animal Nutrition and Feeding Department of Animal Nutrition and Feed ScienceCollege of Animal Science and TechnologyChina Agricultural UniversityBeijing 100193People’s Republic of China

Background Vitamin A(VA)and its metabolite,retinoic acid(RA),are of great interest for their wide range of physiological functions.However,the regulatory contribution of VA to mitochondrial and muscle fiber composition in sheep has not been reported.Method Lambs were injected with 0(control)or 7,500 IU VA palmitate into the biceps femoris muscle on d 2 after birth.At the age of 3 and 32 weeks,longissimus dorsi(LD)muscle samples were obtained to explore the effect of VA on myofiber type composition.In vitro,we investigated the effects of RA on myofiber type composition and intrinsic mechanisms.Results The proportion of type I myofiber was greatly increased in VA-treated sheep in LD muscle at harvest.VA greatly promoted mitochondrial biogenesis and function in LD muscle of sheep.Further exploration revealed that VA elevated PGC-1αmRNA and protein contents,and enhanced the level of p38 MAPK phosphorylation in LD muscle of sheep.In addition,the number of type I myofibers with RA treatment was significantly increased,and type IIx myofibers was significantly decreased in primary myoblasts.Consistent with in vivo experiment,RA significantly improved mitochondrial biogenesis and function in primary myoblasts of sheep.We then used si-PGC-1αto inhibit PGC-1αexpression and found that si-PGC-1αsignificantly abrogated RA-induced the formation of type I myofibers,mitochondrial biogenesis,MitoTracker staining intensity,UQCRC1 and ATP5A1 expression,SDH activity,and enhanced the level of type IIx muscle fibers.These data suggested that RA improved mitochondrial biogenesis and function by promoting PGC-1αexpression,and increased type I myofibers.In order to prove that the effect of RA on the level of PGC-1αis caused by p38 MAPK signaling,we inhibited the p38 MAPK signaling using a p38 MAPK inhibitor,which significantly reduced RA-induced PGC-1αand MyHC I levels.Conclusion VA promoted PGC-1αexpression through the p38 MAPK signaling pathway,improved mitochondrial biogenesis,and altered the composition of muscle fiber type.

关键词： Mitochondria Muscle fiber type PGC-1α p38 MAPK Retinoic acid Vitamin A

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Probiotic cocktails accelerate baicalin metabolism in the ileum to modulate intestinal health in broiler chickens

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Journal of animal science and Biotechnology 2024年第2期15卷 779-794页

作者： Mingkun Gao Chaoyong Liao Jianyang Fu Zhonghua Ning Zengpeng Lv Yuming Guo State Key Laboratory of animal Nutrition and Feeding College of Animal Science and TechnologyChina Agricultural UniversityBeijing 100193China National Engineering Laboratory for animal Breeding College of Animal Science and TechnologyChina Agricultural UniversityBeijing 100193China

Background Baicalin and probiotic cocktails are promising feed additives with broad application prospects.While probiotic cocktails are known to enhance intestinal health,the potential synergistic impact of combining baicalin with probiotic cocktails on the gut health of broiler chickens remains largely unexplored.Therefore,this study aims to investigate the influence of the combined administration of baicalin and probiotic cocktails on the composition of ileal and cecal microbiota in broiler chickens to elucidate the underlying mechanisms responsible for the healthpromoting effects.Results A total of 3201-day-old male Arbor Acres broilers were divided into 4 groups,each with 8 replicates of 10 chicks per replicate.Over a period of 42 d,the birds were fed a basal diet or the same diet supplemented with 37.5 g/t baicalin(BC),1,000 g/t probiotic cocktails(PC),or a combination of both BC(37.5 g/t)and PC(1,000 g/t).The results demonstrated that BC+PC exhibited positive synergistic effects,enhancing intestinal morphology,immune function,and barrier function.This was evidenced by increased VH/CD ratio,sIgA levels,and upregulated expression of occludin and claudin-1(P<0.05).16S rRNA analysis indicated that PC potentiated the effects of BC,particularly in the ileum,where BC+PC significantly increased theα-diversity of the ileal microbiota,altered itsβ-diversity,and increased the relative abundance of Flavonifractor(P<0.05),a flavonoid-metabolizing bacterium.Furthermore,Flavonifractor positively correlated with chicken ileum crypt depth(P<0.05).While BC+PC had a limited effect on cecal microbiota structure,the PC group had a very similar microbial composition to BC+PC,suggesting that the effect of PC at the distal end of the gut overshadowed those of BC.Conclusions We demonstrated the synergistic enhancement of gut health regulation in broiler chickens by combining baicalin and probiotic cocktails.Probiotic cocktails enhanced the effects of baicalin and accelerated its metabolism in the ileum,thereby influencing the ileal

关键词： Baicalin Gut health Ileal microbiota Probiotic cocktails Synergistic effects

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Dietary copper requirement of broilers fed a corn-soybean meal diet during 22—42 d of age

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animal Nutrition 2024年第1期16卷 96-104页

作者： Ling Zhu Wei Wu Bingxin Wu Yun Hu Liyang Zhang Weiyun Zhang Tingting Li Xiaoyan Cui Feiyu Gao Ding Li Xugang Luo Shengchen Wang Poultry Mineral Nutrition Laboratory College of Animal Science and TechnologyYangzhou University Mineral Nutrition Research Division State Key Laboratory of Animal NutritionInstitute of Animal ScienceChinese Academy of Agricultural Sciences

This research was to assess the dietary copper(Cu) requirement of broiler chickens fed a practical cornsoybean meal diet during 22—42 d of age. A total of 288 numbered Arbor Acres male broilers at 22 d of age were randomly allotted 6 treatments with 8 replicate cages(6 broilers per cage) per treatment.Broilers were fed a Cu-unsupplemented corn-soybean meal basal diet(control, containing 7.36 mg Cu/kg) or the basal diet added with 3, 6, 9, 12, or 15 mg Cu/kg from CuSO4·5H2O for 21 d. Quadratic,asymptotic and broken-line models were fitted and the best fitted models were selected to determine dietary Cu requirements. The results revealed that the contents of Cu in serum and liver, m RNA expression levels of Cu-and zinc-containing superoxide dismutase(CuZnSOD) in liver and monoamine oxidase b(MAO B) in heart, as well as protein expression level of CuZnSOD in liver were affected(P < 0.05) by supplemental Cu levels, and the above indices varied linearly and quadratically(P < 0.05)with increasing Cu levels. Dietary Cu requirements assessed according to the best fitted broken-line models(P < 0.05) of the above indexes were 10.45—13.81 mg/kg. It was concluded that mRNA expression levels of CuZnSOD in liver and MAO B in heart, as well as liver CuZnSOD protein expression level were new specific sensitive biomarkers for estimating dietary Cu requirements, and the dietary Cu requirement was recommended to be 14 mg/kg to support Cu metabolic needs related to key Cucontaining enzymes in broilers fed the corn-soybean meal diet during 22—42 d of age, which was higher than the dietary Cu requirement(8 mg/kg) for broilers at the corresponding stage suggested by the Chinese Feeding Standard of Chicken.

关键词： Copper Copper content Gene expression of copper-containing enzyme Requirement Broiler

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Associations of genome-wide structural variations with phenotypic differences in cross-bred Eurasian pigs

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Journal of animal science and Biotechnology 2024年第1期15卷 68-87页

作者： Wencheng Zong Jinbu Wang Runze Zhao Naiqi Niu Yanfang Su Ziping Hu Xin Liu Xinhua Hou Ligang Wang Lixian Wang Longchao Zhang State Key Laboratory of animal Biotech Breeding Institute of Animal SciencesChinese Academy of Agricultural SciencesBeijing 100193China College of animal science Shanxi Agricultural UniversityJinzhong 030801China College of animal science and Technology Qingdao Agricultural UniversityQingdao 266109China

Background During approximately 10,000 years of domestication and selection,a large number of structural variations(SVs)have emerged in the genome of pig breeds,profoundly influencing their phenotypes and the ability to adapt to the local environment.SVs(≥50 bp)are widely distributed in the genome,mainly in the form of insertion(INS),mobile element insertion(MEI),deletion(DEL),duplication(DUP),inversion(INV),and translocation(TRA).While studies have investigated the SVs in pig genomes,genome-wide association studies(GWAS)-based on SVs have been rarely conducted.Results Here,we obtained a high-quality SV map containing 123,151 SVs from 15 Large White and 15 Min pigs through integrating the power of several SV tools,with 53.95%of the SVs being reported for the first time.These high-quality SVs were used to recover the population genetic structure,confirming the accuracy of genotyping.Potential functional SV loci were then identified based on positional effects and breed stratification.Finally,GWAS were performed for 36 traits by genotyping the screened potential causal loci in the F2 population according to their corresponding genomic positions.We identified a large number of loci involved in 8 carcass traits and 6 skeletal traits on chromosome 7,with FKBP5 containing the most significant SV locus for almost all traits.In addition,we found several significant loci in intramuscular fat,abdominal circumference,heart weight,and liver weight,etc.Conclusions We constructed a high-quality SV map using high-coverage sequencing data and then analyzed them by performing GWAS for 25 carcass traits,7 skeletal traits,and 4 meat quality traits to determine that SVs may affect body size between European and Chinese pig breeds.

关键词： Body size GWAS Pig Skeleton Structural variations

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Ginkgo biloba extract alleviates fatty liver hemorrhagic syndrome in laying hens via reshaping gut microbiota

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Journal of animal science and Biotechnology 2024年第1期15卷 277-294页

作者： Xinyue Yang Depeng Li Meihong Zhang Yuqing Feng Xiaolu Jin Dan Liu Yuming Guo Yongfei Hu State Key Laboratory of animal Nutrition College of Animal Science and TechnologyChina Agricultural UniversityBeijing 100193China

Background Ginkgo biloba extract(GBE)is evidenced to be effective in the prevention and alleviation of metabolic disorders,including obesity,diabetes and fatty liver disease.However,the role of GBE in alleviating fatty liver hemorrhagic syndrome(FLHS)in laying hens and the underlying mechanisms remain to be elucidated.Here,we investigated the effects of GBE on relieving FLHS with an emphasis on the modulatory role of GBE in chicken gut microbiota.Results The results showed that GBE treatment ameliorated biochemical blood indicators in high-fat diet(HFD)-induced FLHS laying hen model by decreasing the levels of TG,TC,ALT and ALP.The lipid accumulation and pathological score of liver were also relieved after GBE treatment.Moreover,GBE treatment enhanced the antioxidant activity of liver and serum by increasing GSH,SOD,T-AOC,GSH-PX and reducing MDA,and downregulated the expression of genes related to lipid synthesis(FAS,LXRα,GPAT1,PPARγand Ch REBP1)and inflammatory cytokines(TNF-α,IL-6,TLR4 and NF-κB)in the liver.Microbial profiling analysis revealed that GBE treatment reshaped the HFD-perturbed gut microbiota,particularly elevated the abundance of Megasphaera in the cecum.Meanwhile,targeted metabolomic analysis of SCFAs revealed that GBE treatment significantly promoted the production of total SCFAs,acetate and propionate,which were positively correlated with the GBE-enriched gut microbiota.Finally,we confirmed that the GBE-altered gut microbiota was sufficient to alleviate FLHS by fecal microbiota transplantation(FMT).Conclusions We provided evidence that GBE alleviated FLHS in HFD-induced laying hens through reshaping the composition of gut microbiota.Our findings shed light on mechanism underlying the anti-FLHS efficacy of GBE and lay foundations for future use of GBE as additive to prevent and control FLHS in laying hen industry.

关键词： Antioxidation Fecal microbiota transplantation FLHS Ginkgo biloba extract Inflammation Megasphaera

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Dietary supplementation of laminarin improves the reproductive performance of sows and the growth of suckling piglets

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Journal of animal science and Biotechnology 2024年第1期15卷 329-343页

作者： Pengguang Zhang Guoyuan Jiang Chenghong Ma Yubo Wang Enfa Yan Linjuan He Jianxin Guo Xin Zhang Jingdong Yin State Key Laboratory of animal Nutrition and Feeding College of Animal Science and TechnologyChina Agricultural UniversityBeijing 100193China

Background Maternal nutrition is essential in keeping a highly efficient production system in the pig industry.Laminarin has been shown to improve antioxidant capacity,reduce the inflammatory response,and favor the homeostasis of intestinal microbiota.However,the effect of dietary supplementation of laminarin on the reproductive performance of sows and the growth of suckling offspring remains unknown.Methods A total of 40 Landrace×Yorkshire multiparous sows on d 85 of gestation,similar in age,body weight(BW),parity and reproductive performance,were randomly divided into four dietary treatments with 10 sows per treatment,receiving a control diet(basal pregnancy or lactating diets)and a basal diet supplemented with 0.025%,0.05%and 0.10%laminarin,respectively.The experiment lasted from d 85 of gestation to d 21 of lactation.Results Laminarin supplementation linearly increased number born alive per litter(P=0.03),average daily feed intake(ADFI,P<0.01),and total milk yield of sows during the lactation of 1–21 d(P=0.02).Furthermore,maternal laminarin supplementation increased the average daily gain(ADG)of piglets while tending to reduce the culling and death rate before weaning.In addition,alterations to the composition of colostrum and milk,as well as to serum inflammatory cytokines and immunoglobulins of sows were observed.The fecal microbiota profile of sows supported the improvement of reproductive performance in sows and the growth performance in suckling offspring.Conclusions Dietary supplementation of laminarin during late pregnancy and lactation could significantly improve reproductive performance of sows and growth performance of piglets.

关键词： Fecal microbiota Laminarin Milk Piglets Reproductive performance Sows

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

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Journal of animal science and Biotechnology 2024年第2期15卷 826-843页

作者： Shengchen Wang Bingxin Wu Ling Zhu Weiyun Zhang Liyang Zhang We Wu Jiaqi Wu Yun Hu Tingting Li Xiaoyan Cui Xugang Luo College of animal science and Technology Yangzhou UniversityYangzhou 225000People’s Republic of China Mineral Nutrition Research Division State Key Laboratory of Animal NutritionInstitute of Animal ScienceChinese Academy of Agricultural SciencesBeijing 100193China

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

关键词： Broilers Chelation strengths Fe-containing enzymes NaFeEDTA Relative bioavailabilities

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Complexed amino acid minerals vs. bis-glycinate chelated minerals:Impact on the performance of old laying hens

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animal Nutrition 2024年第1期16卷 395-408页

作者： Marcos J.B.Santos Maria C.M.M.Ludke Leandro M.Silva Carlos B.V.Rabello Mércia R.Barros Fabiano S.Costa Clariana S.Santos Jamille S.S.Wanderley Department of animal science Rural Federal University of Pernambuco Department of Veterinary science Rural Federal University of Pernambuco

The present study was to evaluate the effect of trace minerals(Zn, Mn, and Cu) from complexed amino acid minerals(ZMCAA) and bis-glycinate chelated minerals(ZMCGly) in laying hen diets on performance,internal and external egg quality, yolk mineral deposition, intestinal morphometry, and bone characteristics. From 78 to 98 weeks of age, 400 White LSL-Lite strain laying hens were distributed in a randomized design with 4 treatments with 10 replicates per treatment. Treatments were distributed in a 2 ×2 factorial arrangement using either Zn, Mn, and Cu of ZMCAA or ZMCGly source at 2 levels: low(20, 20,and 3.5 mg/kg of Zn, Mn, and Cu, respectively) or high(40, 40, and 7 mg/kg of Zn, Mn, and Cu,respectively). The analysis of variance was performed, and in cases where differences were observed, the means were compared using Tukey?s test(P < 0.05). The source and level of trace mineral supplementation had a significant impact on the performance of laying hens. Hens fed ZMCAA had higher egg production(P = 0.01), egg weight(P = 0.02), egg mass(P = 0.01), and lower feed conversion ratio(P =0.05) compared to those fed ZMCGly. The ZMCAA supplementation showed higher albumen height(P =0.01), albumen weight(P = 0.01), and eggshell thickness(P < 0.01). The deposition of Zn(P < 0.01), Mn(P< 0.01), and Cu(P < 0.01) in the egg yolk was greater for hens received ZMCAA. Tibia weight(P = 0.04)and bone densitometry(P < 0.01) in the tibia were higher with ZMCAA supplementation. In the small intestine, ZMCAA resulted in longer villi(P = 0.02) and shorter crypt depth(P = 0.01) in the duodenum.Jejunum and ileum measurements were influenced by the level and source of trace minerals(P < 0.05).Laying hens fed ZMCAA exhibited superior performance, egg quality, deposition of trace minerals in the egg yolk, and bone density compared to hens fed ZMCGly. In this study, older laying hens supplemented with ZMCAA at lower levels demonstrated adequate levels of supplementation.

关键词： Bis-glycinates Metal-amino acids Radiodensity Trace minerals

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