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The early life immune dynamics and cellular drivers at single-cell resolution in lamb forestomachs and abomasum

Journal of Animal Science and Biotechnology 2024年第1期15卷 218-235页

作者： Kailang Huang Bin Yang Zebang Xu Hongwei Chen Jiakun Wang Institute of Dairy Science College of Animal SciencesZhejiang UniversityHangzhou 310058China Key Laboratory of Molecular Animal Nutrition Ministry of EducationZhejiang UniversityHangzhou 310058China

Background Four-chambered stomach including the forestomachs(rumen,reticulum,and omasum)and abomasum allows ruminants convert plant fiber into high-quality animal products.The early development of this four-chambered stomach is crucial for the health and well-being of young ruminants,especially the immune development.However,the dynamics of immune development are poorly understood.Results We investigated the early gene expression patterns across the four-chambered stomach in Hu sheep,at 5,10,15,and 25 days of age.We found that forestomachs share similar gene expression patterns,all four stomachs underwent widespread activation of both innate and adaptive immune responses from d 5 to 25,whereas the metabolic function were significantly downregulated with age.We constructed a cell landscape of the four-chambered stomach using single-cell sequencing.Integrating transcriptomic and single-cell transcriptomic analyses revealed that the immune-associated module hub genes were highly expressed in T cells,monocytes and macrophages,as well as the defense-associated module hub genes were highly expressed in endothelial cells in the four-stomach tissues.Moreover,the non-immune cells such as epithelial cells play key roles in immune maturation.Cell communication analysis predicted that in addition to immune cells,non-immune cells recruit immune cells through macrophage migration inhibitory factor signaling in the forestomachs.Conclusions Our results demonstrate that the immune and defense responses of four stomachs are quickly developing with age in lamb's early life.We also identified the gene expression patterns and functional cells associated with immune development.Additionally,we identified some key receptors and signaling involved in immune regulation.These results help to understand the early life immune development at single-cell resolution,which has implications to develop nutritional manipulation and health management strategies based on specific targets including key receptors and signaling pathways.

关键词： Early life Forestomachs Four-chambered stomach Immune cells Immune system maturation MIF signaling rumen Ruminant development Single-cell transcriptomic sequencing

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rumen microbial-driven metabolite from grazing lambs potentially regulates body fatty acid metabolism by lipid-related genes in liver

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Journal of Animal Science and Biotechnology 2023年第4期14卷 1678-1695页

作者： Zhen Li Xingang Zhao Luyang Jian Bing Wang Hailing Luo State Key Laboratory of Animal Nutrition College of Animal Science and TechnologyChina Agricultural UniversityBeijing 100193P.R.China

Background Lipid metabolism differs significantly between grazing and stall-feeding lambs,affecting the quality of livestock products.As two critical organs of lipid metabolism,the differences between feeding patterns on rumen and liver metabolism remain unclear.In this study,16S rRNA,metagenomics,transcriptomics,and untargeted metabolomics were utilized to investigate the key rumen microorganisms and metabolites,as well as liver genes and metabolites associated with fatty acid metabolism under indoor feeding(F)and grazing(G).Results Compared with grazing,indoor feeding increased ruminal propionate content.Using metagenome sequencing in combination with 16S rRNA amplicon sequencing,the results showed that the abundance of propionate-producing Succiniclasticum and hydrogenating bacteria Tenericutes was enriched in the F group.For rumen metabolism,grazing caused up-regulation of EPA,DHA and oleic acid and down-regulation of decanoic acid,as well as,screening for 2-ketobutyric acid as a vital differential metabolite,which was enriched in the propionate metabolism pathway.In the liver,indoor feeding increased 3-hydroxypropanoate and citric acid content,causing changes in propionate metabolism and citrate cycle,while decreasing the ETA content.Then,the liver transcriptome revealed that 11 lipid-related genes were differentially expressed in the two feeding patterns.Correlation analysis showed that the expression of CYP4A6,FADS1,FADS2,ALDH6A1 and CYP2C23 was significantly associated with the propionate metabolism process,suggesting that propionate metabolism may be an important factor mediating the hepatic lipid metabolism.Besides,the unsaturated fatty acids in muscle,rumen and liver also had a close correlation.Conclusions Overall,our data demonstrated that rumen microbial-driven metabolite from grazing lambs potentially regulates multiple hepatic lipid-related genes,ultimately affecting body fatty acid metabolism.

关键词： Feeding pattern Lamb Lipid metabolism Liver Microorganism rumen

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The role of rumen epithelial urea transport proteins in urea nitrogen salvage:A review

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Animal Nutrition 2022年第2期9卷 304-313页

作者： Chongliang Zhong Ruijun Long Gavin S.Stewart College of Ecology State Key Laboratory of Grassland Agro-ecosystemsLanzhou UniversityLanzhouChina School of Biology and Environmental Science University College DublinDublinIreland

The symbiotic relationship between the host and the rumen microbiome plays a crucial role in ruminant physiology.One of the most important processes enabling this relationship is urea nitrogen salvaging(UNS).This process is important for both maintaining ruminant nitrogen balance and supporting production of their major energy supply,bacterially-derived short chain fatty acids(SCFA).The key step in UNS is the trans-epithelial movement of urea across the ruminal wall and this is a highly regulated process.At the molecular level,the key transport route is via the facilitative urea transporter-B2,localized to ruminal papillae epithelial layers.Additional urea transport through aquaporins(AQP),such as AQP3,is now also viewed as important.Long-term regulation of these ruminal urea transport proteins appears to mainly involve dietary fermentable carbohydrates;whereas,transepithelial urea transport is finely regulated by local conditions,such as CO_(2) levels,pH and SCFA concentration.Although the key principles of ruminal urea transport physiology are now understood,there remains much that is unknown regarding the regulatory pathways.One reason for this is the limited number of techniques currently used in many studies in the field.Therefore,future research in this area that combines a greater range of techniques could facilitate improvements to livestock efficiency,and potentially,reductions in the levels of waste nitrogen entering the environment.

关键词： Urea nitrogen salvaging rumen Urea transporter Aquaporin UT-B AQP

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Single-cell transcriptomic landscape of the sheep rumen provides insights into physiological programming development and adaptation of digestive strategies

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Zoological Research 2022年第4期43卷 634-647页

作者： Yuan Yuan Da-Ming Sun Tao Qin Sheng-Yong Mao Wei-Yun Zhu Yu-Yang Yin Jie Huang Rasmus Heller Zhi-Peng Li Jun-Hua Liu Qiang Qiu School of Ecology and Environment Northwestern Polytechnical UniversityXi’anShaanxi 710072China Ruminant Nutrition and Feed Engineering Technology Research Center College of Animal Science and TechnologyNanjing Agricultural UniversityNanjingJiangsu 210095China Laboratory of Gastrointestinal Microbiology Jiangsu Key Laboratory of Gastrointestinal Nutrition and Animal HealthNational Center for International Research on Animal Gut NutritionCollege of Animal Science and TechnologyNanjing Agricultural UniversityNanjingJiangsu 210095China Huzhou Academy of Agricultural Sciences HuzhouZhejiang 313000China Section for Computational and RNA Biology Department of BiologyUniversity of CopenhagenCopenhagen DK-2200Denmark College of Animal Science and Technology Jilin Agricultural UniversityChangchunJilin 130118China

As an important evolutionary innovation and unique organ,the rumen has played a crucial role in ruminant adaptation to complex ecological environments.However,the cellular basis of its complex morphology and function remains largely unknown.In this study,we identified eight major cell types from seven representative prenatal and postnatal rumen samples using~56600 single-cell transcriptomes.We captured the dynamic changes and high heterogeneity in cellular and molecular profiles before,during,and after the appearance of keratinized stratified squamous epithelium with neatly arranged papillae and functional maturity.Basal cells,keratinocytes,differentiating keratinocytes,terminally differentiated keratinocytes,and special spinous cells provided the cellular basis for rumen epithelium formation.Notably,we obtained clear evidence of two keratinization processes involved in early papillogenesis and papillae keratinization and identified TBX3 as a potential marker gene.Importantly,enriched stratum spinosum cells played crucial roles in volatile fatty acid(VFA)metabolism and immune response.Our results provide a comprehensive transcriptional landscape of rumen development at single-cell resolution,as well as valuable insight into the interactions between dietary metabolism and the rumen.

关键词： rumen scRNA-seq Ruminal epithelium Keratinization

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Antimicrobial peptides act on the rumen the performance of castrated bulls

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Journal of Animal Science and Biotechnology 2023年第3期14卷 1218-1234页

作者： Jinping Shi Yu Lei Jianping Wu Zemin Li Xiao Zhang Li Jia Ying Wang Yue Ma Ke Zhang Qiang Cheng Zhao Zhang Yannan Ma Zhaomin Lei College of Animal Science and Technology Gansu Agricultural UniversityLanzhou 730070China Key Laboratory of Animal Genetics Breeding and Reproduction of Shanxi ProvinceCollege of Animal Science and TechnologyNorthwest A&F UniversityYangling 712100China Institute of Rural Development Northwest Normal UniversityLanzhou 730070China Jingchuan Xu Kang Food Co. Ltd.Pingliang 744300China Gansu Huarui Agriculture Co. Ltd.Zhangye 734500China

Background Many countries have already banned the use of antibiotics in animal husbandry,making it extremely difficult to maintain animal health in livestock breeding.In the livestock industry,there is an urgent need to develop alternatives to antibiotics which will not lead to drug resistance on prolonged use.In this study,eighteen castrated bulls were randomly divided into two groups.The control group(CK)was fed the basal diet,while the antimicrobial peptide group(AP)was fed the basal diet supplemented with 8 g of antimicrobial peptides in the basal diet for the experimental period of 270 d.They were then slaughtered to measure production performance,and the ruminal contents were isolated for metagenomic and metabolome sequencing analysis.Result The results showed that antimicrobial peptides could improve the daily weight,carcass weight,and net meat weight of the experimental animals.Additionally,the rumen papillae diameter and the micropapillary density in the AP were significantly greater than those in the CK.Furthermore,the determination of digestive enzymes and fermentation parameters showed that the contents of protease,xylanase,andβ-glucoside in the AP were greater than those in the CK.However,lipase content in the CK was greater than that in the AP.Moreover,the content of acetate,propionate,butyrate,and valerate was found to be greater in AP than those in CK.The metagenomic analysis annotated 1993 differential microorganisms at the species level.The KEGG enrichment of these microorganisms revealed that the enrichment of drug resistance-related pathways was dramatically decreased in the AP,whereas the enrichment of immune-related pathways was significantly increased.There was also a significant reduction in the types of viruses in the AP.187 probiotics with significant differences were found,135 of which were higher in AP than in CK.It was also found that the antimicrobial mechanism of the antimicrobial peptides was quite specific.Seven low-abundance microorganisms(Acinetobactersp.Ac1271,Aequorivita soeso

关键词： Antimicrobial peptides Castrated bull Growth performance Metabolites Microorganisms rumen

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Transcriptomic and metabolomic insights into the roles of exogenous β-hydroxybutyrate acid for the development of rumen epithelium in young goats

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Animal Nutrition 2023年第4期15卷 10-21页

作者： Yimin Zhuang Jianmin Chai Mahmoud M.Abdelsattar Yuze Fu Naifeng Zhang Key Laboratory of Feed Biotechnology of the Ministry of Agriculture and Rural Affairs Institute of Feed Research of Chinese Academy of Agricultural SciencesBeijing 100081China Department of Animal Science Division of AgricultureUniversity of ArkansasFayettevilleAR 72701USA Guangdong Provincial Key Laboratory of Animal Molecular Design and Precise Breeding College of Life Science and EngineeringFoshan UniversityFoshanChina Department of Animal and Poultry Production Faculty of AgricultureSouth Valley University83523 QenaEgypt

Beta-hydroxybutyric acid(BHBA),as one of the main metabolic ketones in the rumen epithelium,plays critical roles in cellular growth and metabolism.The ketogenic capacity is associated with the maturation of rumen in young ruminants,and the exogenous BHBA in diet may promote the rumen development.However,the effects of exogenous BHBA on rumen remain unknown.This is the first study to investigate the mechanisms of BHBA on gene expression and metabolism of rumen epithelium using young goats as a model through multi-omics techniques.Thirty-two young goats were divided into control,low dose,middle dose,and high dose groups by supplementation of BHBA in starter(0,3,6,and 9 g/day,respectively).Results demonstrated the dietary of BHBA promoted the growth performance of young goats and increased width and length of the rumen papilla(Prumen characteristics and BHBA concentration were identified.Several upregulated hub genes including NDUFC1,NDUFB4,NDUFB10,NDUFA11 and NDUFA1 were enriched in the gene ontology(GO)pathway of nicotinamide adenine dinucleotide(NADH)dehydrogenase(ubiquinone)activity,while ATP5ME,ATP5PO and ATP5PF were associated with ATP synthesis.RT-PCR revealed the expression of genes(HMGCS2,BDH1,SLC16A3,etc.)associated with lipolysis increased significantly by BHBA supplementation(Prumen development through altering NADH balance and accelerating lipid metabolism,which provides a theoretical guidance for the strategies of gastrointestinal health and development of young ruminants.

关键词： rumen Beta-hydroxybutyrate acid Lipid metabolism Transcriptome Metabolomics Goat

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Localization of urea transporter B in the developing bovine rumen

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Animal Nutrition 2022年第3期10卷 216-222页

作者： Chongliang Zhong Tamsin Lyons Orla Heussaff Evelyn Doyle Eoin O'Hara Sinead M.Waters David Kenny Gavin S.Stewart School of Biology and Environmental Science University College DublinBelfieldDublin 4Ireland Animal and Bioscience Research Department Animal and Grassland Research and Innovation CentreTeagascGrangeCounty MeathIreland Department of Agriculture Foodand Nutritional SciencesUniversity of AlbertaEdmontonAlbertaCanada

Urea nitrogen secreted from blood to rumen is a crucial factor shaping the symbiotic relationship between host ruminants and their microbial populations.Passage of urea across rumen epithelia is facilitated by urea transporter B(UT-B),but the long-term regulation of these proteins remains unclear.As ruminal function develops over a period of months,the developing rumen is an excellent model with which to investigate this regulation.Using rumen epithelium samples of calves from birth to 96 d of age,this study performed immunolocalization studies to localize and semi-quantify UT-B protein development.As expected,preliminary experiments confirmed that ruminal monocarboxylate transporter 1(MCT1)short chain fatty acid transporter protein abundance increased with age(Prumen function after the advent of solid feed intake and weaning,exhibiting a similar pattern to both MCT1 transporters and papillae growth.This study therefore confirmed age-dependent changes of in situ ruminal UT-B protein,adding to our understanding of the long-term regulation of ruminal urea transporters.

关键词： UT-B Immunolocalization rumen Bovine Development

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Lovastatin as a supplement to mitigate rumen methanogenesis:an overview

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Journal of Animal Science and Biotechnology 2022年第2期13卷 353-364页

作者： Amaury Ábrego-Gacía Héctor M.Poggi-Varaldo Vania Robles-González Teresa Ponce-Noyola Graciano Calva-Calva Elvira Ríos-Leal Daniel Estrada-Bárcenas Alfredo Mendoza-Vargas Department of Biotechnology and Bioengineering CINVESTAV-IPNP.O.Box 17-74007000 Mexico CityMexico Environmental Biotechnology and Renewable Energies Group CINVESTAV-IPNP.O.Box 17-74007000 Mexico CityMexico Instituto de Hidrología Universidad Tecnológica de la MixtecaOaxaca 69000 Huajuapan de LeónMexico National Collection of Microbial and Cellular Cultures CINVESTAV-IPNP.O.Box17-74007000 Mexico CityMexico Unidad de Secuenciación e Identificación de Polimorfismos Instituto Nacional de Medicina Genómica14610 Mexico CityMexico

Methane from enteric fermentation is the gas with the greatest environmental impact emitted by ruminants.Lovastatin(Lv)addition to feedstocks could be a strategy to mitigate rumen methane emissions via decreasing the population of methanogenic archaea(MA).Thus,this paper provides the first overview of the effects of Lv supplementation,focusing on the inhibition of methane production,rumen microbiota,and ruminal fermentation.Results indicated that Lv treatment had a strong anti-methanogenic effect on pure strains of MA.However,there are uncertainties from in vitro rumen fermentation trials with complex substrates and rumen inoculum.Solid-state fermentation(SSF)has emerged as a cost-effective option to produce Lv.In this way,SSF of agricultural residues as an Lv-carrier supplement in sheep and goats demonstrated a consistent decrease in ruminal methane emissions.The experimental evidence for in vitro conditions showed that Lv did not affect the volatile fatty acids(VFA).However,in vivo experiments demonstrated that the production of VFA was decreased.Lv did not negatively affect the digestibility of dry matter during in vitro and in vivo methods,and there is even evidence that it can induce an increase in digestibility.Regarding the rumen microbiota,populations of MA were reduced,and no differences were detected in alpha and beta diversity associated with Lv treatment.However,some changes in the relative abundance of the microbiota were induced.Further studies are recommended on:(i)Lv biodegradation products and stability,as well as its adsorption onto the solid matter in the rumen,to gain more insight on the“available”or effective Lv concentration;and(ii)to determine whether the effect of Lv on ruminal fermentation also depends on the feed composition and different ruminants.

关键词： Fermentation Lovastatin Methanogenesis Microbiota rumen

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rumen-protected methionine during the peripartal period in dairy cows and its effects on abundance of major species of ruminal bacteria

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Journal of Animal Science and Biotechnology 2018年第2期9卷 460-466页

作者： Mohamed K.Abdelmegeid Ahmed A.Elolimy Zheng Zhou Vincenzo Lopreiato Joshua C.Mc Cann Juan J.Loor Department of Animal Sciences Mammalian Nutri Physio GenomicsUniversity of Illinois Department of Animal Medicine Faculty of Veterinary Medicine Kafrelsheikh University Department of Animal and Veterinary Sciences Clemson University Department of Health Science Interdepartmental Services Centre of Veterinary for Human and Animal Health Magna Gr?cia University of Catanzaro Division of Nutritional Sciences Illinois Informatics Institute University of Illinois

Background: Extensive degradation of amino acids in the rumen via microbial deamination decreases the postruminal availability of dietary indispensable amino acids. Together with the normal decrease in voluntary dry matter intake(DMI) around parturition in dairy cows, microbial metabolism contributes to a markedly negative balance of indispensable amino acids, including methionine which may be the first-limiting for milk production. The main objective of the current study was to profile changes in major bacterial species with key functions in cel ulose and hemicel ulose digestion, xylan breakdown, proteolytic action, propionic acid production, lactate utilization and ruminal biohydrogenation in cows supplemented with rumen-protected Methionine(SM; Smartamine M, Adisseo NA, Alpharetta,GA, USA) from-23 through 30 d relative to parturition. Because ~90% of the methionine in SM bypasses the rumen,~10% of the methionine is released into the rumen and can be utilized by microbes.Results: As expected, there was an increase in overall DMI after parturition(Day, P < 0.05) during which cows consumed on average 19.6 kg/d versus 13.9 kg/d in the prepartum period. The postpartum diet contained greater concentrations of lipid and highly-fermentable carbohydrate from corn grain, which likely explains the increases in the relative abundance of Anaerovibrio lipolytica, Megasphaera elsdenii, Prevotella bryantii, Selenomonas ruminantium,Streptococcus bovis, and Succinimonas amylolytica. Despite similar DMI prepartum, cows fed SM had greater(Treatment × Day, P < 0.05) abundance prepartum of Fibrobacter succinogenes, Succinimonas amylolytica, and Succinivibrio dextrinosolvens. However, the greater DMI in cows fed SM after parturition(19.6 kg/d versus 13.9 kg/d) was associated with lower abundance of Fibrobacter succinogenes(2.13 × 10-3 versus 2.25 × 10-4) and Selenomonas ruminantium(2.98 × 10-1 versus 4.10 × 10-1). A lower abundance(Day, P < 0.05) was detected on d 20 compared with d-10 for Fibrobacter succinogenes and Succinivibrio

关键词： Microbiome rumen microbes Transition cow

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Effects of live yeast on differential genetic and functional attributes of rumen microbiota in beef cattle

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Journal of Animal Science and Biotechnology 2019年第4期10卷 1101-1107页

作者： Ibukun M.Ogunade Jerusha Lay Kenneth Andries Christina J.Mc Manus Frederick Bebe College of Agriculture Communities and the Environment Kentucky State University

Several studies have evaluated the effects of live yeast supplementation on rumen microbial population;however,its effect on differential microbial genes and their functional potential has not been described. Thus, this study applied shotgun metagenomic sequencing to evaluate the effects of live yeast supplementation on genetic and functional potential of the rumen microbiota in beef cattle. Eight rumen-cannulated Holstein steers were randomly assigned to two treatments in a cross-over design with two 25-day experimental periods and a 10-day wash-out between the two periods. The steers were housed in individual pens and fed 50% concentrate-mix and 50% red clover/orchard hay ad libitum. Treatments were(1) control(CON;basal diet without additive) and(2) yeast(YEA;basal diet plus 15 g/d of live yeast product). rumen fluid samples were collected at 3, 6, and 9 h after feeding on the last d of each period. Sequencing was done on an Illumina Hi Seq 2500 platform. Dietary yeast supplementation increased the relative abundance of carbohydrate-fermenting bacteria(such as Ruminococcus albus, R.champanellensis, R. bromii, and R. obeum) and lactate-utilizing bacteria(such as Megasphaera elsdenii, Desulfovibrio desulfuricans, and D. vulgaris). A total of 154 differentially abundant genes(DEGs) were obtained(false discovery rate < 0.01). Kyoto Encyclopedia of Genes and Genomes(KEGG) annotation analysis of the DEGs revealed that 10 pathways, including amino sugar and nucleotide sugar metabolism, oxidative phosphorylation, lipopolysaccharide biosynthesis, pantothenate and coenzyme A biosynthesis, glutathione metabolism, beta-alanine metabolism,polyketide sugar unit biosynthesis, protein export, ribosome, and bacterial secretory system, were enriched in steers fed YEA. Annotation analysis of the DEGs in the carbohydrate-active enzymes(CAZy) database revealed that the abundance of genes coding for enzymes belonging to glycoside hydrolases, glycosyltransferases, and carbohydrate binding modules were enriched in steers fed YEA. Thes

关键词： Functional potential Live yeast rumen

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