动物营养学报 >

2022 , Vol. 34 >Issue 2: 1087 - 1097

DOI: https://doi.org/10.3969/j.issn.1006-267x.2022.02.040

研究论文 RESEARCH PAPER

凉茶渣替代象草对育肥牛空肠组织形态、屏障功能以及菌群结构的影响

  • 李玲 ,
  • 孙小红 ,
  • 连旭 ,
  • 罗君谊 ,
  • 陈婷 ,
  • 习欠云 ,
  • 张永亮 ,
  • 孙加节
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  • 华南农业大学动物科学学院, 广东省动物营养调控重点实验室, 广州 510642
李玲(1997-),女,河南信阳人,硕士研究生,从事动物营养组学研究。E-mail:1350979603@qq.com

收稿日期: 2021-07-14

  网络出版日期: 2022-02-15

基金资助

国家重点研发计划（2018YFD0501700）；广东省重点研发计划（2019B110209005）；广东省现代农业技术体系（2018LM1121，2018LM2158）

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Effects of Herbal Tea Residue on Ileum Morphology, Barrier Function and Flora Structure of Fattening Cattle

  • LI Ling ,
  • SUN Xiaohong ,
  • LIAN Xu ,
  • LUO Junyi ,
  • CHEN Ting ,
  • XI Qianyun ,
  • ZHANG Yongliang ,
  • SUN Jiajie
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  • Key Laboratory of Animal Nutrition Control in Guangdong, College of Animal Science, South China Agricultural University, Guangzhou 510642, China

Received date: 2021-07-14

  Online published: 2022-02-15

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摘要

本试验旨在研究凉茶渣替代象草对育肥牛空肠组织形态、屏障功能以及菌群结构的影响。选取120头健康且体重相近的西门塔尔杂交肉牛，随机分为2个组，分别为对照组（饲喂基础饲粮）、试验组（50％凉茶渣替代部分象草），每组3个重复，每个重复20头。试验期共67 d，包括7 d的预试期，60 d的正试期。在试验期结束后，采集试验牛血液、空肠组织、空肠内容物，通过苏木精-伊红（HE）染色测量空肠绒毛高度、隐窝深度；通过酶联免疫吸附测定（ELISA）试剂盒检测血清中炎症因子、D-乳酸（D-Lac）以及二胺氧化酶（DAO）浓度；通过荧光定量PCR测定空肠紧密连接蛋白基因闭合小环蛋白-1（ZO-1）、闭锁蛋白（Occludin）、闭合蛋白-1（Claudin-1）的mRNA表达水平；利用16S rDNA高通量测序分析空肠菌群结构。结果表明：1）与对照组相比，试验组育肥牛空肠绒毛高度显著增加（P<0.05），血清中D-Lac浓度显著降低（P<0.05），空肠紧密连接蛋白ZO-1的mRNA表达水平显著上调（P<0.05）。2）在门水平上，对照组和试验组厚壁菌门（Firmicutes）相对丰度分别为85.79％、76.96％，拟杆菌门（Bacteroidetes）相对丰度分别为0.54％、3.11％。在属水平上，对照组和试验组之间罗姆布茨菌属（Romboutsia）、罗斯氏菌属（Rothia）、布劳特氏菌属（Blautia）等相对丰度表现出显著差异（P<0.05）。3） Pearson相关性分析表明，空肠绒毛高度与Blautia、Rothia、短波单胞菌属（Brevundimonas）、四球虫属（Tetrasphaera）相对丰度呈显著正相关（P<0.05），相关系数为0.46~0.57；血清中D-Lac浓度与Romboutsia、土孢杆菌属（Terrisporobacter）相对丰度呈显著正相关（P<0.05），相关系数分别为0.56、0.62；空肠紧密连接蛋白ZO-1的mRNA表达水平与梭菌属（Solobacterium）相对丰度呈显著负相关（P<0.05），相关系数为-0.50。由此可见，饲粮中添加50％凉茶渣作为功能性粗饲料对育肥牛空肠绒毛高度、屏障功能有积极的影响，凉茶渣可能通过改变空肠菌群组成和相对丰度，进而影响空肠组织形态与屏障功能，最终促进肠道健康。

关键词： 凉茶渣; 育肥牛; 空肠; 组织形态; 屏障功能; 空肠菌群

本文引用格式

李玲 , 孙小红 , 连旭 , 罗君谊 , 陈婷 , 习欠云 , 张永亮 , 孙加节 . 凉茶渣替代象草对育肥牛空肠组织形态、屏障功能以及菌群结构的影响[J]. 动物营养学报, 2022 , 34(2) : 1087 -1097 . DOI: 10.3969/j.issn.1006-267x.2022.02.040

Abstract

This experiment was conducted to study the effects of herbal tea residue on ileum morphology, barrier function and flora structure of fattening cattle. A total of 120 healthy Simmental crossbred beef cattle with similar body weight were randomly divided into two groups:control group (fed a basal diet) and experimental group (50% herbal tea residue replaced part of elephant grass), with 3 replicates in each group and 20 heads in each replicate. The experimental period lasted for 67 days, including 7 days of pre feeding period and 60 days of formal experimental period. At the end of the experimental period, the blood, jejunal tissue and jejunal contents were collected, and the villus height and crypt depth of jejunum were measured by hematoxylin-eosin (HE) staining; the concentrations of inflammatory factors, D-lactic acid (D-Lac) and diamine oxidase (DAO) in serum were measured by enzyme-linked immunosorbent assay (ELISA) kit; the mRNA expression levels of tight junction proteins of zonula occludens-1 (ZO-1), Occludin and Claudin-1 in jejunum were determined by fluorescence quantitative PCR; the 16S rDNA high-throughput sequencing was used to analyze the structure of jejunal flora. The results showed as follows:1) compared with the control group, the jejunal villi height of the experimental group was significantly increased (P<0.05), the serum D-Lac concentration was significantly decreased (P<0.05), and the mRNA expression level of tight junction protein ZO-1 in jejunum was significantly up-regulated (P<0.05). 2) At the phylum level, the Firmicutes relative abundances in the control group and the experimental group were 85.79% and 76.96%, respectively, and the Bacteroidetes relative abundances were 0.54% and 3.11%, respectively. At the genus level, the relative abundances of Romboutsia, Rothia and Blautia showed significant differences between the control group and the experimental group (P<0.05). 3) Pearson correlation analysis displayed that the jejunal villi height showed significant positive correlations with the relative abundances of Blautia, Brevundimonas, Rothia and Tetrasphaera (P<0.05), and the correlation coefficients ranged from 0.46 to 0.57; the serum D-Lac concentration showed significant positive correlations with the relative abundances of Romboutsia and Terrisporobacter (P<0.05), and the correlation coefficients were 0.56 and 0.62, respectively; the mRNA expression level of tight junction protein ZO-1 in jejunum showed a significant negative correlation with the relative abundances of Solobacterium (P<0.05), and correlation coefficient was -0.50. In conclusion, adding 50% herbal tea residue as functional roughage in the diet has positive effects on the jejunal villus height and barrier function of fattening cattle, the herbal tea residue may affect intestinal tissue morphology and barrier function by changing the composition and relative abundance of jejunal flora, and ultimately promote the intestinal health.

Key words： herbal tea residue; fattening cattle; jejunum; tissue morphology; barrier function; jejunal flora

参考文献

[1] 刘主洁.王老吉凉茶的化学成分分析及质量评价研究[D].硕士学位论文.广州:广州中医药大学, 2016. LIU Z J.Chemical components analysis and quality control of Wanglaoji herbal tea[D].Master's Thesis.Guangzhou:Guangzhou University of Chinese Medicine, 2016.(in Chinese)
[2] 贺颖颖, 罗燕玉, 林朝展, 等.王老吉凉茶化学成分研究[J].中药材, 2018, 41(4):889-893. HE Y Y, LUO Y Y, LIN C Z, et al.Chemical constituents from Wanglaoji herbal tea[J].Journal of Chinese Medicinal Materials, 2018, 41(4):889-893.(in Chinese)
[3] LI D L, ZHENG X L, DUAN L, et al.Ethnobotanical survey of herbal tea plants from the traditional markets in Chaoshan, China[J].Journal of Ethnopharmacology, 2017, 205:195-206.
[4] RAHMAN I U, AFZAL A, IQBAL Z, et al.Herbal teas and drinks:folk medicine of the Manoor Valley, Lesser Himalaya, Pakistan[J].Plants, 2019, 8(12):581.
[5] AJILA C M, BRAR S K, VERMA M, et al.Bio-processing of agro-byproducts to animal feed[J].Critical Reviews in Biotechnology, 2012, 32(4):382-400.  
[6] XIE Y Q, CHEN Z J, WANG D Y, et al.Effects of fermented herbal tea residues on the intestinal microbiota characteristics of Holstein heifers under heat stress[J].Frontiers in Microbiology, 2020, 11:1014.
[7] 张萌萌, 姜宁, 张爱忠.肠道微生物对肠道屏障功能完整性的维护机制研究概况[J].微生物学通报, 2020, 47(3):933-940. ZHANG M M, JIANG N, ZHANG A Z.Maintenance mechanism of intestinal barrier function integrity by intestinal microbes[J].Microbiology, 2020, 47(3):933-940.(in Chinese)
[8] 武笑天, 温曌, 李锐瑞, 等.叶酸和维生素B12对产蛋鸡生产性能、小肠屏障功能及相关基因表达的影响[J].动物营养学报, 2021, 33(5):2616-2630. WU X T, WEN Z, LI R R, et al.Effects of folic acid and vitamin B12 on performance, small intestinal barrier function and related gene expression of laying hens[J].Chinese Journal of Animal Nutrition, 2021, 33(5):2616-2630.(in Chinese)
[9] PAULSON J N, POP M, BRAVO H C.Metastats:an improved statistical method for analysis of metagenomic data[J].Genome Biology, 2011, 12(Suppl.1):P17.
[10] CHAPMAN M G, UNDERWOOD A J.Ecological patterns in multivariate assemblages:information and interpretation of negative values in ANOSIM tests[J].Marine Ecology Progress Series, 1999, 180:257-265.
[11] 陈耀星, 崔燕.动物解剖学与组织胚胎学[M].北京:中国农业出版社, 2019. CHEN Y X, CUI Y.Animal anatomy and histoembryology[M].Beijing:China Agriculture Press, 2019.(in Chinese)
[12] 王娇, 杨宝钰, 阿依古丽·艾买尔, 等.甜高粱与苜蓿混合青贮对肉羊消化道组织形态的影响[J].饲料工业, 2021, 42(7):54-60. WANG J, YANG B Y, AIMAIER A Y G L, et al.Effects of sweet sorghum and alfalfa silage mixtures on digestive tissue morphology in meat sheep[J].Feed Industry, 2021, 42(7):54-60.(in Chinese)
[13] WOYENGO T A, KIARIE E, NYACHOTI C M.Metabolizable energy and standardized ileal digestible amino acid contents of expeller-extracted canola meal fed to broiler chicks[J].Poultry Science, 2010, 89(6):1182-1189.  
[14] 王凤英.中草药结合α-酮戊二酸对断奶大鼠消化道生长发育的影响[D].硕士学位论文.乌鲁木齐:新疆农业大学, 2004. WANG F Y.The effects of Chinese herbs combined with α-ketoglutarate on the small intestinal morphology and physiology in weaned rats[D].Master's Thesis.Urumqi:Xinjiang Agricultural University, 2004.(in Chinese)
[15] CHEN J, GUO K, SONG X Z, et al.The anti-heat stress effects of Chinese herbal medicine prescriptions and rumen-protected γ-aminobutyric acid on growth performance, apparent nutrient digestibility, and health status in beef cattle[J].Animal Science Journal, 2020, 91(1):e13361.
[16] HUANG S W, FU Y J, XU B, et al.Wogonoside alleviates colitis by improving intestinal epithelial barrier function via the MLCK/pMLC2 pathway[J].Phytomedicine, 2020, 68:153179.
[17] ZEISEL M B, DHAWAN P, BAUMERT T F.Tight junction proteins in gastrointestinal and liver disease[J].Gut, 2019, 68(3):547-561.  
[18] KAWANO M, MIYOSHI M, OGAWA A, et al.Lactobacillus gasseri SBT2055 inhibits adipose tissue inflammation and intestinal permeability in mice fed a high-fat diet[J].Journal of Nutritional Science, 2016, 5:e23.
[19] GAO M, JIANG Y, XIAO X F, et al.Protective effect of pioglitazone on sepsis-induced intestinal injury in a rodent model[J].The Journal of Surgical Research, 2015, 195(2):550-558.  
[20] DUZGUN A P, GULGEZ B, OZMUTLU A, et al.The relationship between intestinal hypoperfusion and serum d-lactate levels during experimental intra-abdominal hypertension[J].Digestive Diseases and Sciences, 2006, 51(12):2400-2403.  
[21] HONZAWA Y, NAKASE H, MATSUURA M, et al.Clinical significance of serum diamine oxidase activity in inflammatory bowel disease:importance of evaluation of small intestinal permeability[J].Inflammatory Bowel Diseases, 2011, 17(2):E23-E25.
[22] SU J Y, ZHU Q, ZHAO Y, et al.Dietary supplementation with Chinese herbal residues or their fermented products modifies the colonic microbiota, bacterial metabolites, and expression of genes related to colon barrier function in weaned piglets[J].Frontiers in Microbiology, 2018, 9:3181.
[23] SHREINER A B, KAO J Y, YOUNG V B.The gut microbiome in health and in disease[J].Current Opinion in Gastroenterology, 2015, 31(1):69-75.  
[24] MA Q T, LI Y Q, LI P F, et al.Research progress in the relationship between type 2 diabetes mellitus and intestinal flora[J].Biomedicine & Pharmacotherapy, 2019, 117:109138.
[25] MA N, TIAN Y A, WU Y, et al.Contributions of the interaction between dietary protein and gut microbiota to intestinal health[J].Current Protein & Peptide Science, 2017, 18(8):795-808.
[26] SARESELLA M, MENDOZZI L, ROSSI V, et al.Immunological and clinical effect of diet modulation of the gut microbiome in multiple sclerosis patients:a pilot study[J].Frontiers in Immunology, 2017, 8:1391.
[27] WANG H, JI Y C, YIN C, et al.Differential analysis of gut microbiota correlated with oxidative stress in sows with high or low litter performance during lactation[J].Frontiers in Microbiology, 2018, 9:1665.
[28] GAGEN E J, PADMANABHA J, DENMAN S E, et al.Hydrogenotrophic culture enrichment reveals rumen Lachnospiraceae and Ruminococcaceae acetogens and hydrogen-responsive Bacteroidetes from pasture-fed cattle[J].FEMS Microbiology Letters, 2015, 362(14):fnv104.
[29] NAKAYAMA J, YAMAMOTO A, PALERMO-CONDE L A, et al.Impact of westernized diet on gut microbiota in children on Leyte island[J].Frontiers in Microbiology, 2017, 8:197.
[30] 许杰, 王伊龙, 王拥军.肠道菌群与高血压[J].中国卒中杂志, 2017, 12(2):175-178. XU J, WANG Y L, WANG Y J.Gut microbiome and hypertension[J].Chinese Journal of Stroke, 2017, 12(2):175-178.(in Chinese)
[31] LI J, ZHAO F Q, WANG Y D, et al.Gut microbiota dysbiosis contributes to the development of hypertension[J].Microbiome, 2017, 5(1):14.
[32] TURNBAUGH P J, LEY R E, MAHOWALD M A, et al.An obesity-associated gut microbiome with increased capacity for energy harvest[J].Nature, 2006, 444(7122):1027-1031.  
[33] PLAIZIER J C, LI S, TUN H M, et al.Nutritional models of experimentally-induced subacute ruminal acidosis (SARA) differ in their impact on rumen and hindgut bacterial communities in dairy cows[J].Frontiers in Microbiology, 2017, 7:2128.
[34] 孙鹏.奶牛酮病的发生原因、临床特征、诊断及防治措施[J].现代畜牧科技, 2021(3):78-79. SUN P.Causes, clinical features, diagnosis and control measures of ketosis in dairy cows[J].Modern Animal Husbandry Science & Technology, 2021(3):78-79.(in Chinese)
[35] BERNI CANANI R, SANGWAN N, STEFKA A T, et al.Lactobacillus rhamnosus GG-supplemented formula expands butyrate-producing bacterial strains in food allergic infants[J].The ISME Journal, 2016, 10(3):742-750.  
[36] WEI G X, DARWISH G, OPPENHEIM F G, et al.Commensal bacterium Rothia aeria degrades and detoxifies gluten via a highly effective subtilisin enzyme[J].Nutrients, 2020, 12(12):3724.
[37] MANGIFESTA M, MANCABELLI L, MILANI C, et al.Mucosal microbiota of intestinal polyps reveals putative biomarkers of colorectal cancer[J].Scientific Reports, 2018, 8(1):13974.
[38] KELLER M, MAZUCH J, ABRAHAM U, et al.A circadian clock in macrophages controls inflammatory immune responses[J].Proceedings of the National Academy of Sciences of the United States of America, 2009, 106(50):21407-21412.  
[39] 尉浩斌, 张小丹, 李苏宜.肠屏障功能损伤机制及其临床检测方法研究现状[J].肿瘤代谢与营养电子杂志, 2020, 7(4):407-414. WEI H B, ZHANG X D, LI S Y.Research status of the mechanism of intestinal barrier function injury and its clinical detection methods[J].Electronic Journal of Metabolism and Nutrition of Cancer, 2020, 7(4):407-414.(in Chinese)
[40] BACH KNUDSEN K E, LÆRKE H N, HEDEMANN M S, et al.Impact of diet-modulated butyrate production on intestinal barrier function and inflammation[J].Nutrients, 2018, 10(10):1499.
[41] CAI C X, ZHANG Z X, MORALES M, et al.Feeding practice influences gut microbiome composition in very low birth weight preterm infants and the association with oxidative stress:a prospective cohort study[J].Free Radical Biology & Medicine, 2019, 142:146-154.
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