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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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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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Biotic and abiotic stresses on honeybee health

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Integrative Zoology 2024年第3期19卷 442-457页

作者： Zheguang LIN Siyi SHEN Kang WANG Ting JI Apicultural Research institute College of Animal Science and TechnologyYangzhou UniversityYangzhouChina

Honeybees are the most critical pollinators providing key ecosystem services that underpin crop production and sustainable agriculture.Amidst a backdrop of rapid global change,this eusocial insect encounters a succession of stressors during nesting,foraging,and pollination.Ectoparasitic mites,together with vectored viruses,have been recognized as central biotic threats to honeybee health,while the spread of invasive giant hornets and small hive beetles also increasingly threatens colonies worldwide.Cocktails of agrochemicals,including acaricides used for mite treatment,and other pollutants of the environment have been widely documented to affect bee health in various ways.Additionally,expanding urbanization,climate change,and agricultural intensification often result in the destruction or fragmentation of flower-rich bee habitats.The anthropogenic pressures exerted by beekeeping management practices affect the natural selection and evolution of honeybees,and colony translocations facilitate alien species invasion and disease transmission.In this review,the multiple biotic and abiotic threats and their interactions that potentially undermine bee colony health are discussed,while taking into consideration the sensitivity,large foraging area,dense network among related nestmates,and social behaviors of honeybees.

关键词： abiotic stressor Apis biotic stressor colony losses interaction effect pollinator decline

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Importance of dietary supplementation of soluble and insoluble fibers to sows subjected to high ambient temperatures during late gestation and effects on lactation performance

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animal Nutrition 2024年第1期16卷 73-83页

作者： Seung Min Oh Abdolreza Hosseindoust Sang Hun Ha Jun Young Mun Joseph Moturi Habeeb Tajudeen Yo Han Choi Su Hyup Lee Jin Soo Kim Gyeongbuk Livestock Research institute Department of animal Industry Convergence Kangwon National University Swine Division National Institute of Animal Science Rural Development Administration Department of Swine science Korea National University of Agriculture and Fisheries

Heat stress adversely affects sows' performance, which can be improved by applying proper nutritional strategies. This study was conducted to investigate the interactive effects of dietary fiber levels and sources on sows' reproductive performance, metabolic response during gestation, and the carry-over influence on litter performance in the lactation period during heat stress(average room temperature of 27.1°C). Fifty-four multiparous sows(Landrace × Yorkshire; initial body weight of 236.3 ± 16 kg; 2, 3and 4 parities) at d 90 of gestation were assigned to a 2 × 3 factorial arrangement(9 sows/treatment),involving 2 dietary fiber levels(4.5% and 6% crude fiber) and 3 dietary fiber sources(wheat bran [WB],palm kernel meal [PK], and beet pulp [BP]). Sows fed the BP diet had highest(P < 0.01) feed intake and constipation index and lowest(P < 0.01) farrowing duration. Piglet weight(P = 0.041) and litter weight(P < 0.01) at weaning were higher in sows in the BP treatment compared to PK treatment. Sows in the BP treatment showed the greatest(P < 0.01) digestibility of crude protein and neutral detergent fiber. The fecal concentration of acetate was the lowest(P < 0.01) in the PK treatment. Total short-chain fatty acid production was increased in the WB and BP treatments compared with the PK. Sows in the BP treatment showed the lowest(P = 0.036) hair cortisol. The blood insulin concentration of sows was higher(P = 0.026) in the high fiber(6%) treatment compared with the low fiber(4.5%) treatment at 90 min and120 min after the meal. The concentration of phthalic acid, succinic acid, phenylethylamine, hydrocinnamic acid, iron, linoleic acid, glycerol, ketone, and formamide were increased(P < 0.05) in the BP treatment compared with the WB. The BP treatment with high soluble fiber content improved the constipation index, farrowing duration, and litter performance, while high insoluble fibers increased sows comfort and reduced stress factors including respiratory rate and rectal temperature. Therefore,both soluble and insolub

关键词： Soluble fiber Insoluble fiber Beet pulp Wheat bran Palm kernel Metabolites

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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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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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Mulberry leaf supplementation inhibits skatole deposition by regulating gut microbiota and upregulating liver cytochrome P4501A1 expression in finishing pigs

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animal Nutrition 2024年第1期16卷 34-44页

作者： Yuqing Sun Xiaoming Men Tianbao Lin Bo Deng Shi Zhong Jinxi Huo Kaipeng Qin Zhiqiang Lv Ziwei Xu Yougui Li institute of Sericulture and Tea Zhejiang Academy of Agricultural Sciences institute of animal Husbandry and Veterinary science Zhejiang Academy of Agricultural Sciences

Skatole, a strong fecal odor substance, is generated through microbial degradation of tryptophan in the animal hindgut. It easily accumulates in adipose tissue and affects meat quality. In this study, the effect of mulberry leaf supplementation on skatole in finishing pigs was studied. In a 35-day trial, 20 finishing pigs (barrows and gilts) were fed with a basal diet or basal diet with 6% mulberry leaves. Growth performance of the pigs (n=10) was automatically recorded by a performance-testing feeder system and 8 pigs in each treatment were slaughtered and sampled for the remaining tests. Skatole and short-chain fatty acids were detected using HPLC and gas chromatography, respectively. Fecal microbiota were analyzed using 16S r RNA gene sequencing. The metabolomics analysis of feces and serum was performed with UHPLC-MS/MS. The major cytochrome P450 (CYP) enzymes that catalyze skatole degradation in the liver were tested by using RT-PCR and Western blot. Effects of major bioactive compounds in mulberry leaves on the CYP genes were verified in the hepatic cell line Hep G2 in an in vitro test (n=3). In finishing pigs, mulberry leaf supplementation had no significant effect on the average daily gain, average daily feed intake, and feed conversion ratio (P>0.05), but reduced skatole levels in feces, serum, and backfat (P<0.05), and increased acetic acid levels in feces (P=0.027). Mulberry leaf supplementation decreased the relative abundance of the skatole-producing bacteria Megasphaera and Olsenella (P<0.05). Indole-3acetic acid, the intermediate that is essential for skatole production, was significantly reduced in feces by mulberry leaf supplementation (P<0.05) and was positively correlated with skatole content in feces (P=0.004). In pigs treated with mulberry leaves, liver CYP1A1 expression was increased (P<0.05) and was negatively correlated with skatole content in backfat (P=0.045). The in vitro test demonstrated that mulberry leaf polyphenols and polysaccharides could directly stimulate CYP1A1 expression in h

关键词： Pig Skatole Mulberry leaf Microbiota Cytochrome P450

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Selenomethionine alleviates chronic heat stress-induced breast muscle injury and poor meat quality in broilers via relieving mitochondrial dysfunction and endoplasmic reticulum stress

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animal Nutrition 2024年第1期16卷 363-375页

作者： Jinzhong Jing Jiayi Wang Xiaoyu Xiang Shenggang Yin Jiayong Tang Longqiong Wang Gang Jia Guangmang Liu Xiaoling Chen Gang Tian Jingyi Cai Bo Kang Lianqiang Che Hua Zhao Key Laboratory for animal Disease-Resistance Nutrition of Ministry of Education of China Ministry of Agriculture and Rural Affairsof Sichuan ProvinceAnimal Nutrition InstituteSichuan Agricultural University College of animal science and Technology Sichuan Agricultural University

In the present study,the chronic heat stress （CHS） broiler model was developed to investigate the potential protection mechanism of organic selenium （selenomethionine,SeMet） on CHS-induced skeletal muscle growth retardation and poor meat quality.Four hundred Arbor Acres male broilers （680±70 g,21 d old） were grouped into 5 treatments with 8 replicates of 10 broilers per replicate.Broilers in the control group were raised in a thermoneutral environment （22±2°C） and fed with a basal diet.The other four treatments were exposed to hyperthermic conditions （33±2°C,24 h in each day） and fed on the basal diet supplied with SeMet at 0.0,0.2,0.4,and 0.6 mg Se/kg,respectively,for 21 d.Results showed that CHS reduced （P<0.05） the growth performance,decreased （P<0.05） the breast muscle weight and impaired the meat quality of breast muscle in broilers.CHS induced protein metabolic disorder in breast muscle,which increased （P<0.05） the expression of caspase 3,caspase 8,caspase 9 and ubiquitin proteasome system related genes,while decreased the protein expression of P-4EBP1.CHS also decreased the antioxidant capacity and induced mitochondrial stress and endoplasmic reticulum （ER） stress in breast muscle,which increased （P<0.05） the ROS levels,decreased the concentration of ATP,increased the protein expression of HSP60 and CLPX,and increased （P<0.05） the expression of ER stress biomarkers.Dietary SeMet supplementation linearly increased （P<0.05） breast muscle Se concentration and exhibited protective effects via up-regulating the expression of the selenotranscriptome and several key selenoproteins,which increased （P<0.05） body weight,improved meat quality,enhanced antioxidant capacity and mitigated mitochondrial stress and ER stress.What's more,SeMet suppressed protein degradation and improved protein biosynthesis though inhibiting the caspase and ubiquitin proteasome system and promoting the mTOR-4EBP1 pathway.In conclusion,dietary SeMet supplementation increases the expression of several key selenoproteins,alleviates mitochondr

关键词： Broiler Chronic heat stress Skeletal muscle Meat quality Mitochondrial dysfunction Endoplasmic reticulum stress

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Imprinting at the KBTBD6 locus involves species-specific m ternal methylation and monoallelic expression in livestock animals

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Journal of animal science and Biotechnology 2024年第1期15卷 127-145页

作者： Jinsoo Ahn In-Sul Hwang Mi-Ryung Park Seongsoo Hwang Kichoon Lee Department of animal sciences The Ohio State UniversityColumbusOH 43210USA animal Biotechnology Division Rural Development AdministrationNational Institute of Animal ScienceJeonbuk 55365Republic of Korea Columbia Center for Translational Immunology Columbia University Irving Medical CenterColumbia UniversityNew YorkNY 10032USA animal Welfare Research Team National Institute of Animal ScienceRDA1500Kongjwipatjwi‑roJeollabuk‑do 55365Republic of Korea

Background The primary differentially methylated regions(DMRs) which are maternally hypermethylated serve as imprinting control regions(ICRs) that drive monoallelic gene expression, and these ICRs have been investigated due to their implications in mammalian development. Although a subset of genes has been identified as imprinted, in-depth comparative approach needs to be developed for identification of species-specific imprinted genes. Here, we examined DNA methylation status and allelic expression at the KBTBD6 locus across species and tissues and explored potential mechanisms of imprinting.Results Using whole-genome bisulfite sequencing and RNA-sequencing on parthenogenetic and normal porcine embryos, we identified a maternally hypermethylated DMR between the embryos at the KBTBD6 promoter Cp G island and paternal monoallelic expression of KBTBD6. Also, in analyzed domesticated mammals but not in humans, non-human primates and mice, the KBTBD6 promoter Cp G islands were methylated in oocytes and/or allelically methyl-ated in tissues, and monoallelic KBTBD6 expression was observed, indicating livestock-specific imprinting. Further analysis revealed that these Cp G islands were embedded within transcripts in porcine and bovine oocytes which coexisted with an active transcription mark and DNA methylation, implying the presence of transcription-dependent imprinting.Conclusions In this study, our comparative approach revealed an imprinted expression of the KBTBD6 gene in domesticated mammals, but not in humans, non-human primates, and mice which implicates species-specific evolution of genomic imprinting.

关键词： Differentially methylated region Domesticated mammal Imprinting KBTBD6 Parthenogenetic

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