人工饲粮对幼年工蜂生长发育及生理代谢的影响

本试验利用液相色谱-质谱联用技术(LC-MS)对幼年工蜂进行代谢组学分析,旨在探究不同人工饲粮对幼年工蜂生长发育及生理代谢的影响,为蜂王邮寄的人工饲粮配制提供一定理论依据。试验选取1日龄的意大利蜜蜂(Apis mellifera ligustica)300只,分为5组,分别饲喂5种人工饲粮,每组30只工蜂。对照组饲粮:蔗糖粉∶蜂蜜=3∶1;试验组A饲粮:蔗糖粉∶蜂粮∶蜂蜜=6∶5∶1;试验组B饲粮:蔗糖粉∶蜂粮∶蜂蜜=6∶3∶1;试验组C饲粮:蔗糖粉∶花粉∶蜂蜜=6∶5∶1;试验组D饲粮:蔗糖粉∶花粉∶蜂蜜=6∶3∶1。用5种人工饲粮进行室内喂养,记录蜜蜂7 d内的死亡数量;解剖试验组A和对照组第1、3、5、7天的工蜂咽下腺,测定其平均重量、颜色和饱满度;采用LC-MS方法检测试验组A和对照组蜜蜂饲喂7 d后的代谢物差异,对检测数据进行模式识别分析,筛选并鉴别差异代谢物。结果表明:1)对照组和试验组A蜜蜂第1~5天时均没有出现死亡,在第6和7天时试验组A蜜蜂死亡数量均显著低于对照组(P<0.05)。试验组B、C、D的蜜蜂在6 d内全部死亡。2)试验组A第7天时的蜜蜂咽下腺平均重量显著高于对照组(P<0.05),咽下腺颜色为全部乳白色,且咽下腺小体饱满。3)采用主成分分析(PCA)、偏最小二乘法判别方法(PLS-DA)和正交偏最小二乘方-判别分析(OPLS-DA)分析代谢组数据,结果显示试验组A和对照组明显分离,共鉴定出23个差异代谢物,包括氨基酸、脂类、糖类等,其中有10种代谢物上调,13种代谢物下调。差异代谢物通路分析发现,差异极显著的通路有牛磺酸和亚牛磺酸生物合成通路、赖氨酸降解通路、戊糖磷酸盐代谢通路、戊糖和葡萄糖醛酸相互转化通路(P<0.01),差异显著的通路有氨基酸的生物合成通路,色氨酸代谢通路,碳代谢通路,丁酸代谢通路,丙氨酸、天冬氨酸和谷氨酸代谢通路,丙酮酸代谢通路(P<0.05)。由此可见,试验组A的饲粮更适合蜜蜂工蜂的生长发育和生存,在邮寄环境中可代替常用的炼糖饲粮。

Effects of Artificial Diets on Growth and Development and Physiological Metabolism of Young Worker Bees

To study the effects of different artificial diets on growth and development and physiological metabolism of young worker bees,the metabonomics analysis was conducted for worker bees based on liquid chromatography-mass spectrometry(LC-MS),and to provide some theoretical basis for the preparation of artificial diet of mailed queen bee.A total of 300 one-day old Apis mellifera ligustica was selected and divided to 5 groups with 30 worker bees in each group,and fed with Five kinds of artificial diets.Control group diet:sucrose powder∶honey=3∶1;experimental group A diet∶sucrose powder∶bee feed∶honey=6∶5∶1;experimental group B diet:sucrose powder∶bee feed∶honey=6∶3∶1;experimental group C diet:sucrose powder∶pollen∶honey=6∶5∶1;experimental group D diet:sucrose powder∶pollen∶honey=6∶3∶1.Five kinds of artificial diet were used for indoor feeding,recorded the death number of honeybees within 7 days.The hypopharyngeal glands of worker bees on days 1,3,5 and 7 were dissected and measured the average weighing,color and plumpness.The metabolite differences of honeybees after feeding 7 days between experimental group A and control group were detected by LC-MS method,the detected data were carry out the pattern recognition analysis,screened and identified the different metabolites.The results showed as follows:1)the honeybees in control group and experimental group A were no death during days 1 to 5,the death number of honeybees on days 6 and 7 of experimental group A was significantly lower than that of the control group(P<0.05).The honeybees in experimental groups B,C and D were all died within 6 days.2)The mean weight of hypopharyngeal gland of honeybees on day 7 of experimental group A was significantly higher than that of the control group(P<0.05),the hypopharyngeal gland colour were all milky white,and hypopharyngeal gland corpuscle was plump.3)The metabolomic data were analyzed by principal component analysis(PCA),partial leastsquares discriminant analysis(PLS-DA)and orthogonal partial leastsquares-discriminant analysis(OPLS-DA),the results showed that the experimental group A and the control group were significantly separated,the 23 different metabolites were identified,included amino acid,lipid and saccharides,among then,10 metabolites were up-regulated and 13 metabolites were down-regulated.The differential metabolite pathways analysis showed that the pathways with significant differences were taurine and hypotaurine metabolism pathway,alysine degradation pathway,pentose phosphate metabolism pathway and pentose and glucuronate interconversions pathway(P<0.01),and the pathways with significant differences were biosynthesis of amino acids pathway,tryptophan metabolism pathway,carbon metabolism pathway,butanoate metabolism pathway,alanine,aspartate and glutamate metabolism pathway and pyruvate metabolism pathway(P<0.05).In conclusion,the diet of experimental group A is more suitable for the growth,development and survival of worker bees,it can replace the common sugar refining diet in mailing environment.