代乳粉对早期断奶沂蒙黑山羊羔小肠发育、菌群多样性及葡萄糖转运载体基因表达的影响
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摘要
【目的】研究饲喂代乳粉对早期断奶沂蒙黑山羊羔小肠发育、菌群多样性及葡萄糖转运载体基因表达的影响,为沂蒙黑山羊早期断奶羔羊的培育提供理论依据。【方法】将36只双胎沂蒙黑山羊羔羊分成2组,每组6个重复,每个重复3只。其中,对照组(Ⅰ组)羔羊随母哺乳至75日龄,试验组(Ⅱ组)于10日龄断奶后饲喂代乳粉。分别于试验的第10、15、25、45和75天称羔羊活体重以及屠宰后十二指肠、空肠、回肠重量,并收集各肠段内容物、黏膜组织样品。采用16S r DNA的PCR-DGGE技术和实时荧光定量PCR等方法,分析小肠菌群定植规律及葡萄糖转运载体(SGLT-1、GLUT-2)基因m RNA表达量。【结果】Ⅱ组羔羊45 d时平均日增重和空肠、回肠重量显著高于Ⅰ组(P<0.05),而75 d时平均日增重和回肠重量极显著高于Ⅰ组(P<0.01)。十二指肠在45和75 d、空肠在15 d及回肠在10 d时两组间菌群变化较为明显,二者菌群多样性指数回肠最高,其次空肠、十二指肠;两组各肠段优势菌群均为溶淀粉乳杆菌和普雷沃氏菌属,但随着日龄增加,Ⅰ组各肠段益生菌属总细菌数量呈下降趋势,Ⅱ组却与此相反。Ⅱ组在10 d时SGLT-1和GLUT-2基因m RNA表达量最低,而25 d后Ⅱ组表达量均高于Ⅰ组,且差异不显著(P>0.05);上述基因表达量呈现肠段部位间差异,其中SGLT-1基因m RNA表达量变化空肠最大,十二指肠最低,而GLUT-2基因m RNA表达量变化十二指肠最大,回肠最低。【结论】饲喂代乳粉在第25天时可促进沂蒙黑山羊早期断奶羔羊空肠、回肠有益菌提前定植,并通过调控葡萄糖转运载体基因表达而引起葡萄糖吸收转运的改变,进而影响羔羊后期肠道组织及机体的生长发育。
【Objective】 This study was conducted to evaluate the effects of the milk replacer on the intestinal development, the flora diversity and the relative expression of glucose transporter gene of Yimeng black goat lambs in order to provide theoretical basis for the cultivation of early stage Yimeng black goat lambs. 【Method】 A total of 36 twin born Yimeng black goat lambs were randomly divided into 2 groups, each with 6 replicates and each replicate 3 lambs. The lambs in the control group(group I) were fed with breast milk for 75 days, while the lambs in the experimental group(group Ⅱ) were weaned on the 10 th day and fed with milk replacer thereafter. The weight of duodenum, jejunum, ileum as well as live body of the lambs in the two groups were recorded on the 10, 15, 25, 45 and 75 th day, and meanwhile, the small intestinal contents and mucous tissue samples were collected. The colonization of small intestinal flora and the expression of glucose transporter(SGLT-1, GLUT-2) gene m RNA were analyzed by the PCR-DGGE of 16 S r DNA and real-time fluorescence quantitative PCR, respectively. 【Result】 The results showed that the average daily gain and the weight of jejunum and ileum of the lambs in group on the 45 th day were significantly higher than those in group I(P<0.05), while on the 75 th day, the average daily gain and ileum weight of the lambs in group were highly significantly higher than those in group I(P<0.01). The flora distribution difference between group I and group was significant in the duodenum on the 45 and 75 th day, in the jejunum on the 15 th day and in the ileum on the 10 th day. The flora diversity in the ileum was higher than that in the jejunum and duodenum, and the microbiota diversity in the jejunum was higher than that in the duodenum. The bacterium lacticum dissolving starch and prevotella were dominant bacteria in the whole intestinal tract of Group I and . The number of probiotics in group I decreased with the growth of age, while an opposite dynamic was found in the group . The expression of SGLT-1 and GLUT-2 genes in the group was the lowest at the 10 d, but their expression increased on the 25 th day, meanwhile the two genes took on different expression profiles in different intestinal segments. The fluctuation of expression level of SGLT-1 was the highest and lowest in the jejunum and duodenum, respectively, while that of GLUT-2 was the biggest and lowest in the duodenum and ileum, respectively.【Conclusion】 In conclusion, feeding milk replacer could promote early colonization of probiotics in the jejunum and ileum of early weaned Yimeng black goat lambs on the 25 th day and the glucose absorption and transport could be changed by regulating glucose transporter gene expression, which further affected later development of intestinal tissue and the growth of the lambs.
【Objective】 This study was conducted to evaluate the effects of the milk replacer on the intestinal development, the flora diversity and the relative expression of glucose transporter gene of Yimeng black goat lambs in order to provide theoretical basis for the cultivation of early stage Yimeng black goat lambs. 【Method】 A total of 36 twin born Yimeng black goat lambs were randomly divided into 2 groups, each with 6 replicates and each replicate 3 lambs. The lambs in the control group(group I) were fed with breast milk for 75 days, while the lambs in the experimental group(group Ⅱ) were weaned on the 10 th day and fed with milk replacer thereafter. The weight of duodenum, jejunum, ileum as well as live body of the lambs in the two groups were recorded on the 10, 15, 25, 45 and 75 th day, and meanwhile, the small intestinal contents and mucous tissue samples were collected. The colonization of small intestinal flora and the expression of glucose transporter(SGLT-1, GLUT-2) gene m RNA were analyzed by the PCR-DGGE of 16 S r DNA and real-time fluorescence quantitative PCR, respectively. 【Result】 The results showed that the average daily gain and the weight of jejunum and ileum of the lambs in group on the 45 th day were significantly higher than those in group I(P<0.05), while on the 75 th day, the average daily gain and ileum weight of the lambs in group were highly significantly higher than those in group I(P<0.01). The flora distribution difference between group I and group was significant in the duodenum on the 45 and 75 th day, in the jejunum on the 15 th day and in the ileum on the 10 th day. The flora diversity in the ileum was higher than that in the jejunum and duodenum, and the microbiota diversity in the jejunum was higher than that in the duodenum. The bacterium lacticum dissolving starch and prevotella were dominant bacteria in the whole intestinal tract of Group I and . The number of probiotics in group I decreased with the growth of age, while an opposite dynamic was found in the group . The expression of SGLT-1 and GLUT-2 genes in the group was the lowest at the 10 d, but their expression increased on the 25 th day, meanwhile the two genes took on different expression profiles in different intestinal segments. The fluctuation of expression level of SGLT-1 was the highest and lowest in the jejunum and duodenum, respectively, while that of GLUT-2 was the biggest and lowest in the duodenum and ileum, respectively.【Conclusion】 In conclusion, feeding milk replacer could promote early colonization of probiotics in the jejunum and ileum of early weaned Yimeng black goat lambs on the 25 th day and the glucose absorption and transport could be changed by regulating glucose transporter gene expression, which further affected later development of intestinal tissue and the growth of the lambs.
引文
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[3]VELAYUDHAN B T,DANIELS K M,HORRELL D P,HILL S R,MCGILLIARD M L,CORL B A,JIANG H,AKERS R M.Developmental histology,segmental expression,and nutritional regulation of somatotropic axis genes in small intestine of pre-weaned dairy heifers.Journal of Dairy Science,2008,91(9):3343-3352.
[4]SUN Z H,HE Z X,ZHANG Q L,TAN Z L,HAN X F,TANG S X,ZHOU C S,WANG M,ZHANG E P.Effects of protein and/or energy restriction for six weeks on antioxidation capacity of Plasma and gastrointestinal epithelial tissues of weaned kids.Livestock Science,2012,149(3):232-241.
[5]MACKIE R I,WHITE B A.Recent advances in rumen microbial ecology and metabolism:Potential impact on nutrient output.Journal of Dairy Science,1990,73(10):2971-2995.
[6]卢玉飞,周凌云,赵圣国,卜登攀,孙鹏,赵国琦.近10年瘤胃微生物分离培养研究进展.中国微生态学杂志,2012,24(9):856-862.LU Y F,ZHOU L Y,ZHAO S G,BU D P,SUN P,ZHAO G Q.Progress on isolation and culture of rumen microorganisms in recent ten years.Chinese Journal of Microecology,2012,24(9):856-862.(in Chinese)
[7]KELLETT G L.The facilitated component of intestinal glucose absorption.Journal of Physiology,2001,531(3):585-595.
[8]梁艳,刘强,王聪,张延利,裴彩霞,王永新,郭刚,霍文婕,张拴林,石彩叶,刘建新.异丁酸对犊牛瘤胃及小肠黏膜基因表达的影响.动物营养学报,2015,27(8):2483-2492.LIANG Y,LIU Q,WANG C,ZHANG Y L,PEI C X,WANG Y X,GUO G,HUO W J,ZHANG S L,SHI C Y,LIU J X.Effects of isobutyrate on gene expressions of ruminal and small intestinal mucosa of calves.Chinese Journal of Animal Nutrition,2015,27(8):2483-2492.(in Chinese)
[9]LIU J,LIU Z,GAO L,CHEN L,ZHANG H.Nutrient-intake-leveldependent regulation of intestinal development in newborn intrauterine growth-restricted piglets via glucagon-like Peptide-2.Animal,2016,10(10):1645-1654.
[10]BRANDSTETTER A M,PFAFFL M W,HOCQUETTE J F,GERRARD D E,PICARD B,GEAY Y,SAUERWEIN H.Effects of muscle type,castration,age,and compensatory growth rate on androgen receptor m RNA expression in bovine skeletal muscle.Journal of Animal Science,2000,78:629-637.
[11]HAN H C,AUSTIN K J,NATHANIELSZ P W,FORD S P,NIJLAND M J,HANSEN T R.Maternal nutrient restriction alters gene expression in the ovine fetal heart.The Journal of Physiology,2004,558(1):111-121.
[12]王建民,曹洪防,王桂芝,谢之景,刘珊珊.山东及国内优良羊品种介绍(下).农业知识:科学养殖,2011:6-12.WANG J M,CAO H F,WANG G Z,XIE Z J,LIU S S.Introduction of fine sheep breeds in Shandong and in domestic(next).Agricultural Knowledge:Scientific Breeding,2011:6-12.(in Chinese)
[13]屠焰,刁其玉,岳喜新.一种0-3月龄羔羊的代乳品及其制备方法:中国,201210365927.6[P].2013.TU Y,DIAO Q Y,YUE X X.The Milk replacer for 0-3 month of lambs and its preparation method:China,201210365927.6[P].2013.(in Chinese)
[14]张丽英.饲料分析及饲料质量检测技术.北京:中国农业大学出版社2007.ZHANG L Y.Feed Analysis and Feed Quality Inspection Technology.Beijing:China Agricultural University Press,2007.(in Chinese)
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