Bacterial community structure in Apis florea larvae analyzed by denaturing gradient gel electrophoresis and 16S rRNA gene sequencing

This study characterizes the colonization and composition of bacterial flora in dwarf Asian honeybee (Apis florea) larvae and compares bacterial diversity and distribution among different sampling locations. A. florea larvae were collected from 3 locations in Chiang Mai province, Thailand. Bacterial DNA was extracted from each larva using the phenol–chloroform method. Denaturing gradient gel electrophoresis was performed, and the dominant bands were excised from the gels, cloned, and sequenced for bacterial species identification. The result revealed similarities of bacterial community profiles in each individual colony, but differences between colonies from the same and different locations. A. florea larvae harbor bacteria belonging to 2 phyla (Firmicutes and Proteobacteria), 5 classes (Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Bacilli, and Clostridia), 6 genera (Clostridium, Gilliamella, Melissococcus, Lactobacillus, Saccharibacter, and Snodgrassella), and an unknown genus from uncultured bacterial species. The classes with the highest abundance of bacteria were Alphaproteobacteria (34%), Bacilli (25%), Betaproteobacteria (11%), Gammaproteobacteria (10%), and Clostridia (8%), respectively. Similarly, uncultured bacterial species were identified (12%). Environmental bacterial species, such as Saccharibacter floricola, were also found. This is the first study in which sequences closely related to Melissococcus plutonius, the causal pathogen responsible for European foulbrood, have been identified in Thai A. florea larvae.     

云南红河和保山两地区小蜜蜂巢房结构比较研究

【目的】本研究旨在对自然环境条件明显不同的云南红河和保山两地区小蜜蜂Apis florea巢房结构进行量化分析,以期揭示这两个地区间小蜜蜂巢房结构的差异性。【方法】测量小蜜蜂子脾厚度、小蜜蜂巢脾上连续10个工蜂巢房的宽度;利用环氧树脂填充巢房,制作巢房模型,基于巢房模型比较两地区小蜜蜂工蜂和雄蜂单个巢房直径、口部边长、棱长、深度和容积及巢房倾斜角的差异。【结果】云南 红河地区小蜜蜂工蜂、雄蜂子脾厚度均极显著小于保山地区的。两地区小蜜蜂连续10个工蜂巢房的宽度在0°方向均显著大于60°和120°方向的宽度,60°和120°方向的宽度之间差异均不显著,两地区小蜜蜂工蜂单个巢房直径在3个方向上也表现出显著差异, 而两地区雄蜂单个巢房的直径在3个方向上均没有显著性差异。红河地区小蜜蜂工蜂巢房直径在3个方向上均显著小于保山地区的,而两地区小蜜蜂雄蜂巢房直径在3个方向上均差异不显著。红河地区小蜜蜂工蜂、雄蜂单个巢房口部边长、棱长、深度和容积均极显著小于保山地区的;两地区工蜂巢房的倾斜角差异不显著。【结论】小蜜蜂巢房的结构存在地区性差异,海拔高、温度低的地区小蜜蜂巢房结构尺寸明显大于海拔低、温度高地区的。本研究结果不仅丰富了小蜜蜂的生物学知识,也为后续研究小蜜蜂巢房结构指标与形态学指标关联性研究提供了思路。     

云南野生小蜜蜂群体SSR遗传多样性分析

【目的】小蜜蜂Apis florea是蜜蜂属野生种类之一,在我国主要分布于西南部。对于小蜜蜂的研究我国多集中在形态方面,遗传多样性的研究较少。本文旨在阐明云南地区小蜜蜂的群体遗传特征,评价其遗传多样性。【方法】本研究利用14对微卫星引物对采自云南省的13群野生小蜜蜂进行了群体遗传变异检测和遗传多样性分析。【结果】在小蜜蜂群体中共检测到53个等位基因,平均等位基因数为3.7857,平均有效等位基因数2.9420,平均期望杂合度和表观杂合度分别为0.5708和0.2184,平均多态信息含量为0.5300。根据Nei’s遗传距离用UPGMA方法对13群小蜜蜂进行亲缘关系分析,结果 13群野生小蜜蜂先分出红河(HH),再分出缅甸(MD)和普洱(PE),最后所有的西双版纳的聚在一起。【结论】说明同一地区的小蜜蜂的遗传距离相对不同地区的较近。     

云南地区小蜜蜂的饲养方法

为了小蜜蜂Apis florea资源的利用和保护,本论文对其饲养方法及生物学特性进行了研究。在云南省蒙自市,收集野生小蜜蜂8群,对其筑巢、蜂蜜生产、蜂群越冬等饲养方法进行研究,观察并记录饲养过程中蜂群生物学特性。辅助蜂群筑巢过程中,支撑巢脾的树枝距离地面高度在0.6-1.0 m,蜂群较稳定,不易弃巢;利用"巢脾分区切割"的方法生产蜂蜜可以提高产量,减少蜂群分蜂次数;模拟野外筑巢环境,用塑料框和稻草包装,置于日光室的网室内,可有效辅助蜂群越冬。通过对小蜜蜂饲养方法的探索,明确了云南地区小蜜蜂饲养过程中的筑巢高度及科学合理的蜂蜜生产、蜂群越冬方法,为深入研究该种传粉蜂资源的利用与保护提供了重要的参考依据。     

膜翅目昆虫酯酶同工酶的比较研究

用聚丙烯酰胺凝胶等电聚焦电泳分析了膜翅目七个总科、蜜蜂总科9个属, 蜜蜂属6个种和西方蜜蜂4个宗样品的酯酶同工酶.总科代表之间酶谱的差别大于总科内各属间的差别;各属间的酶谱也有显著差别;而属内各种间的酶谱比较接近, 但每个种都有自己特有的谱型, 彼此易于区分.没有发现宗间有明显差别.本文还在实验的基础上讨论了用电泳酶谱方法作为昆虫分子分类学研究手段的可行性.     

Essential Oil of Indian Propolis: Chemical Composition and Repellency against the Honeybee Apis florea

Hitherto unknown biological properties and the chemical composition of the essential oil isolated from propolis of Indian origin were established. GC/MS Analysis of the essential oil revealed the presence of 32 constituents, of which ten were major compounds, nine had intermediate contents, and 13 were minor compounds. With the exception of six minor constituents, that could not be identified, their identification was based on the comparison of their mass spectra and Kovats retention indices with those listed in the NIST and Wiley mass spectral libraries. Their structural assignment was confirmed by GC/MS co‐injection of the essential oil with authentic compounds. Quantification of the components was done by GC‐FID analyses. Moreover, the essential oil was shown to possess repellent activity against the honeybee Apis florea. The activity was found to be dose dependent. The average repellency (ΔR) increased with increasing essential‐oil concentration up to 24 μg/ml and remained constant for the formulation with the higher concentration. These findings established the chemical constitution of the essential oil and might be useful to beekeepers for the improvement of the bee management.