不同糖源饲料对越冬蜜蜂肠道健康及肠道微生物区系的影响

发布时间：2018-03-21 05:04

本文选题：越冬蜜蜂　切入点：糖源类型　出处：《山东农业大学》2017年硕士论文　论文类型：学位论文

【摘要】：在我国北方地区,冬季气候寒冷,外界无蜜源,蜜蜂越冬期间无法从外界获取食物,只能消耗越冬前贮备的食物。因此,越冬饲料的质量好坏直接影响着蜂群的健康。自然状态下蜜蜂越冬期的糖饲料是蜜源季节储存的蜂蜜,但是,蜂农为了追求蜂产品产量和利润,往往会在蜜源季取竭蜂群内的储蜜,转而选择廉价的白砂糖作为越冬饲料,更有蜂农甚至会使用更廉价的果葡糖浆替代白砂糖作为越冬饲料。越冬期不同糖源类型对蜜蜂的饲养价值并不清楚,因此,本研究选择蔗糖、蜂蜜、果葡糖浆为蜜蜂越冬期糖源饲料,通过检测越冬期蜜蜂中肠理化指标、组织形态和肠道菌群区系等指标,探索糖源对越冬蜜蜂肠道健康的影响,从而为养蜂生产中越冬饲料的选择提供参考。在秋末(10月底)选取群势相当的本地意大利蜜蜂(Apis mellifera ligustica)越冬蜂群(群内无储备越冬饲料脾)9群,随机分为3个试验组(3群/组),从11月2号开始分别以蜂蜜、白砂糖和果葡糖浆为糖源进行饲喂,至11月下旬喂足越冬饲料,蜂群进入越冬期。分别于越冬前(11月初)、越冬中期(1月初)和越冬后(3月初)采集蜜蜂中肠,测定中肠消化酶(淀粉酶、蔗糖酶和蛋白酶)活性;在越冬中期采集蜜蜂中肠用于中肠组织形态和抗氧化酶基因(Sod1、Sod2和CAT)表达水平等指标的检测;在越冬前(11初)和越冬中期(1月初)采集蜜蜂的中肠和直肠,利用16S rDNA技术测定蜜蜂肠道菌群的种类和丰富度。结果表明:1、不同糖源饲料对越冬蜜蜂中肠消化酶及抗氧化酶基因表达的影响:(1)在越冬中期,果葡糖浆组和蜂蜜组的蜜蜂中肠内淀粉酶活性显著高于蔗糖组(P0.05)。(2)不同糖源饲料对蜜蜂中肠内蔗糖酶活性的影响差异不显著(P0.05)。(3)在越冬中期,果葡糖浆组的蜜蜂中肠内蛋白酶活性显著高于蔗糖组和蜂蜜组(P0.05)。(4)在越冬中期,蔗糖组的蜜蜂中肠Sod1基因的相对表达量显著高于蜂蜜组和果葡糖浆组(P0.05)。2、不同糖源饲料对越冬蜜蜂中肠组织结构的影响:蜂蜜组和蔗糖组的蜜蜂中肠肠壁厚度显著大于果葡糖浆组(P0.01),而且蜂蜜组和蔗糖组的蜜蜂中肠隐窝深度比果葡糖浆组中肠隐窝深度深(P0.01)。3、不同糖源饲料对越冬蜜蜂肠道菌群的种类和丰富度的影响:(1)在门水平,越冬蜜蜂的肠道菌群主要为变形菌门(Proteobacteria,63.17%)、厚壁菌门(Firmicutes,17.61%)、放线菌门(Actinobacteria,4.06%)和拟杆菌门(Bacteroidetes,1.72%)。在属水平,Lactobacillus(15.91%)和Bifidobacterium(3.24%)为主要优势菌属。(2)白砂糖能够提高越冬蜜蜂肠道中放线菌门(Actinobacteria)、双歧杆菌(Bifidobacterium)、α-变形菌纲(Alphaproteobacteria)和根瘤菌目(Rhizobiales)的丰富度,蜂蜜能够提高蜜蜂肠道中γ-变形菌纲(Gammaproteobacteria)和巴斯德氏菌目(Pasteurellales)的丰富度,而果葡糖浆可以增加蜜蜂肠道中β-变形菌纲(Betaproteobacteria)和奈瑟氏菌科(Neisseriaceae)的丰富度。本试验表明,不同糖源饲料能够改变越冬蜜蜂肠道菌群的结构,蜂蜜和白砂糖能够提高越冬蜜蜂肠道中有益菌的相对丰度;蜂蜜能够提高越冬蜜蜂中肠消化酶活性,蜂蜜和白砂糖有利于越冬蜜蜂中肠组织的发育,而且白砂糖能够提高越冬蜜蜂中肠抗氧化基因的表达。因此,蜂蜜和白砂糖比果葡糖浆更适合作为蜜蜂越冬的糖源饲料。
[Abstract]:In the northern area of our country, the cold weather in the winter, there is no nectar, bees during the winter can not get food from the outside, only consumption before the winter supplies of food. Therefore, the quality of winter feed directly affects the health of bees. Sugar feed bee wintering in natural state is seasonal storage of honey honey, but. Beekeepers in pursuit of production and profit of bee products, often take up in a swarm in the honey bee season, opting for cheap white sugar as winter feed, more beekeepers will even use cheaper fructose syrup instead of white sugar as winter feed. Wintering period of different types of sugar source feeding value of bees is not clearly, therefore, the study of sucrose, honey, fructose syrup for bee wintering sugar feed, through the detection of wintering bee midgut physicochemical indexes, morphology and intestinal microflora and other indicators, explore the sugar Of overwintering honey bees intestinal health, so as to provide reference for beekeeping production in winter feed. In late autumn (late October) selected local group of potential of the Italy bee (Apis mellifera ligustica) colony (group without reserve winter winter feeding spleen) were randomly divided into 9 groups, 3 experimental groups (3 groups from the beginning of November 2nd / group), respectively with honey, sugar and fructose syrup for sugar feeding, to late November, enough to feed the feed into the winter, the colony wintering stage. In winter (before the beginning of November), mid winter (early January) and winter (early March) acquisition bee midgut, determination of midgut digestive enzymes (amylase, invertase and protease) activity; in the mid winter collection for bee midgut tissue morphology and midgut antioxidant enzyme genes (Sod1, Sod2 and CAT) to detect the expression level of the index; in the winter before (early 11) and mid winter (early January) acquisition of bees and midgut The rectum, determination of bee species and abundance of intestinal flora by using 16S rDNA technique. The results showed that: 1. The effects of different sugar feed on the expression of overwintering bees of digestive enzymes and antioxidant enzyme genes: (1) in the mid winter fruit starch enzyme activity of glucose syrup and honey bee group group was significantly higher than that of sucrose in the midgut group (P0.05). (2) different sugar feed effects on sucrase activity in the midgut of bees had no significant difference (P0.05). (3) in the mid winter fruit syrup group the protease activity in the midgut of bees and honey group was significantly higher than that of sucrose group (P0.05). (4) in mid winter, relative the expression of Sod1 gene in the bee midgut sucrose group was significantly higher than that of honey group and fructose syrup group (P0.05.2), the influence of different sugar feed on overwintering bee midgut structure: honeybee midgut thickness honey group and sucrose group was significantly greater than that of fructose syrup group (P0.01), and the bee The bee syrup group than the midgut crypt depth of midgut crypt depth honey group and sucrose group (P0.01.3), the deep influence of different glucose feed on overwintering bee species and abundance of intestinal flora: (1) at the gate level, the intestinal flora of overwintering honey bees is mainly Proteobacteria (Proteobacteria, 63.17%). Firmicutes (Firmicutes, 17.61%), actinobacteria (Actinobacteria, 4.06%) and Bacteroidetes (Bacteroidetes, 1.72%). At the genus level, Lactobacillus (15.91%) and Bifidobacterium (3.24%) was the dominant genus. (2) sugar can improve intestinal actinomycetes in overwintering bees (door Actinobacteria), Bifidobacterium (Bifidobacterium), alpha Proteobacteria (Alphaproteobacteria) and rhizobiales (Rhizobiales) abundance, honey bees can improve the intestinal tract gammaproteobacteria (Gammaproteobacteria) and Pasteur's Aphyllophorales (Pasteurellales) abundance, and fruit Glucose syrup can increase the intestinal in beta Proteobacteria (Betaproteobacteria) and Neisseriaceae (Neisseriaceae) richness. The test showed that different glucose feed can change the intestinal flora structure of overwintering bees, honey and white sugar can increase the relative abundance of bacteria in the intestinal tract of overwintering bee honey can improve the enzyme; the activity of midgut digestive overwintering bees, honey and white sugar is conducive to the development of overwintering honey bees in midgut tissues, and sugar can increase the expression of antioxidant genes of the overwintering bee midgut. Therefore, honey and sugar than fruit syrup is more suitable as a source of sugar to feed the bees to overwinter.

【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S894

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