Occurrence and prevalence of seven bee viruses in Apis mellifera and Apis cerana apiaries in China

Populations of Apis mellifera and Apis cerana in China were surveyed for seven bee viruses: acute bee paralysis virus (ABPV), black queen cell virus (BQCV), chronic bee paralysis virus (CBPV), deformed wing virus (DWV), Kashmir bee virus (KBV), sacbrood virus (SBV), and Isreal acute paralysis virus (IAPV). No KBV was detected from any samples of the two species. In A. mellifera, DWV was the most prevalent virus, but in A. cerana, SBV was the dominant. Simultaneous multiple infections of viruses were common in both species. This is the first report of detection of IAPV and CBPV in A. cerana.     

安徽省七种蜜蜂病毒的发生与流行研究

【目的】调查安徽省内7种常见蜜蜂病毒:蜜蜂畸翅病毒(Deformed wing virus,DWV)、以色列急性麻痹病毒(Israeli acute paralysis virus,IAPV)、急性蜜蜂麻痹病毒(Acute bee paralysis virus,ABPV)、慢性麻痹病毒(Chronic bee paralysis virus,CBPV)、黑蜂王台病毒(Black queen cell virus,BQCV)、囊状幼虫病病毒(Sacbrood virus,SBV)、克什米尔病毒(Kashmir bee virus,KBV)的感染发生情况,为安徽养蜂业可持续健康发展提供理论依据。【方法】运用反转录RT-PCR和序列分析比对的方法对安徽省内21个乡镇中的38个蜂场蜜蜂样品进行研究分析,以获得以上7种蜜蜂病毒的特异性发生情况。【结果】意大利蜜蜂Apis mellifera蜂场感染率:DWV(64%),IAPV(43%),CBPV(32%),ABPV(14%),BQCV(11%);中华蜜蜂Apis cerana蜂场感染率:DWV(80%),IAPV(40%),CBPV(30%),ABPV(10%),BQCV(0)。SBV和KBV在所有的蜜蜂样品中均未检测到。【结论】DWV,IAPV,CBPV,ABPV,BQCV在安徽省内大范围都存在发生流行现象,SBV和KBV对安徽蜜蜂的潜在危害可能性小。     

中华蜜蜂工蜂视叶胚后发育过程中的细胞凋亡

本研究通过形态解剖和原位末端标记法(TUNEL),对中华蜜蜂Apis cerana cerana视叶胚后发育过程中的细胞凋亡进行了研究,结果表明:视叶内的细胞程序性死亡开始出现在1龄幼虫末期,随后凋亡细胞数量逐渐增加;在视叶的胚后发育过程中,细胞凋亡经历了3个高峰期,即2龄幼虫、5龄幼虫和蛹发育的第2天;在视叶3个部分的发育中,视髓层中细胞凋亡的数量远远多于视小叶和视神经节层,而视神经节层最少,说明了细胞凋亡的数量和位置与各部分结构发育的时间以及神经投射有关。广泛的细胞凋亡是蜜蜂视叶发育过程中的一个显著特征。     

Associations between reproduction and work in workers of the Asian hive bee Apis cerana

If a honey bee (Apis spp.) colony becomes queenless, about 1/3 of young workers activate their ovaries and produce haploid male-producing eggs. In doing so queenless workers maximize their inclusive fitness because the normal option of vicarious production of relatives via their queen’s eggs is no longer available. But if many workers are engaged in reproduction, how does a queenless colony continue to feed its brood and forage? Here we show that in the Asian hive bee Apis cerana hypopharyngeal gland (HPG) size is larger in queenless workers than in queenright workers and that bees undertaking brood-rearing tasks have larger HPG than same-aged bees that are foraging. In queenless colonies, workers with a smaller number of ovarioles are more likely to have activated ovaries. This reinforces the puzzling observation that a large number of ovarioles reduces reproductive success in queenless A. cerana. It further suggests that reproductive workers either avoid foraging or transition to foraging later in life than non-reproductive workers. Finally, our study also showed that ovary activation and larger-than-average numbers of ovarioles had no statistically detectable influence on foraging specialization for pollen or nectar.     

中国甘肃东北部地区东方蜜蜂(Apis cerana)形态学研究

本研究采集甘肃东北部地区泾川蜂场、庄浪蜂场和平凉蜂场的东方蜜蜂Apis cerana 43群,采样点地貌包括山地、丘陵和平原.运用蜜蜂形态学研究方法对东方蜜蜂40个形态特征指标进行测量分析.运用SPSS13.0对蜜蜂形态数据进行方差分析、主成分分析、聚类分析及环境因子相关性分析.研究结果表明:甘肃东北部地区不同采样点东方蜜蜂形态特征指标存在显著差异,而且与环境因素显著相关.     

Survey of six bee viruses using RT-PCR in Northern Thailand

Six honey bee viruses were surveyed using RT-PCR in Northern Thailand where about 80% of Thai apiaries are located. Tested samples were found to be positive for deformed wing virus (DWV), acute bee paralysis virus (ABPV), sacbrood virus (SBV) and Kashmir bee virus (KBV). In the collected samples, neither chronic bee paralysis virus nor black queen cell virus nucleic acids could be detected. It was found that DWV was the most widespread and ABPV was the second most prevalent. Kashmir bee virus was found only in the Lampang province where high infestation of Varroa destructor mite occurred. Tropilaelaps, European foulbrood, and Chalkbrood diseases were found in some apiaries.     

Horizontal and vertical transmission of viruses in the honey bee, Apis mellifera

The most crucial stage in the dynamics of virus infections is the mode of virus transmission. In general, transmission of viruses can occur through two pathways: horizontal and vertical transmission. In horizontal transmission, viruses are transmitted among individuals of the same generation, while vertical transmission occurs from mothers to their offspring. Because of its highly organized social structure and crowded population density, the honey bee colony represents a risky environment for the spread of disease infection. Like other plant and animal viruses, bee viruses use different survival strategies, including utilization of both horizontal and vertical routes, to transmit and maintain levels in a host population. In this review, we explore the current knowledge about the honey bee viruses and transmission routes of bee viruses. In addition, different transmission strategies on the persistence and dynamics of host-pathogen interactions are also discussed.     

中华蜜蜂雄蜂脑部组织结构观察

通过组织化学方法,对中华蜜蜂Apis cerana cerana Fabricius雄蜂的脑部的形态结构进行观察。结果表明,雄蜂的脑由前脑、中脑和后脑三部分构成,前脑视叶两侧具有大而显著的复眼,而其他结构相对较小。蕈形体在脑中所占的比例小于工蜂和蜂王;中心体的比例小于工蜂,与蜂王接近;中脑的嗅叶相对较为发达,这使它们在同蜂王的交配过程中,能够更灵敏地接收来自于蜂王性外激素的刺激。中华蜜蜂雄蜂的嗅叶具有性二态现象,但是与意大利蜜蜂Apis mellifera L.雄蜂发达的巨大纤维球复合体相比,中华蜜蜂并不明显;除此之外,它们在脑部结构上没有显著差异。     

中华蜜蜂自然分蜂中的亲属优惠行为

以3群中华蜜蜂Apiscerana cerana Fabricius为材料,研究自然分蜂中的亲属优惠行为。分蜂开始后,分别对参与分蜂的工蜂和留在原巢内的工蜂随机取样,用95%酒精保存待用。取下所有王台,放入培养箱中培养出房,每30min观察1次,记录处女王的出房顺序。运用3个MRJP(major royal jelly protein)标记分析蜂群的亲缘关系和亚家系组成。3群实验蜂群分别由13,12和9个亚家系组成。分蜂过程中,王台的全同胞姐妹工蜂绝大多数都选择留在原巢内,说明工蜂能够辨认出全同胞姐妹王台,并且给予优惠。参与分蜂的工蜂和留在原巢内的工蜂间亚家系分布差异极显著,说明一些亚家系分蜂性更强,而另一些亚家系分蜂性较弱。实验首次运用MRJP分子标记技术证明:在自然分蜂过程中,工蜂和王台之间存在亲属优惠行为,并且这种优惠行为可能会刺激其它工蜂产生分蜂趋向性。     

Detection of multiple viruses in queens of the honey bee Apis mellifera L

Individual honey bee Apis mellifera L. queens were examined for the presence of six honey bee viruses including acute bee paralysis virus, chronic bee paralysis virus, black queen cell virus, deformed wing virus, Kashmir bee virus, and sacbrood virus. All viruses, except ABPV, were detected in the samples. Among queens examined for virus infections, 93% had multiple virus infections. The detection of viruses in queens raises the possibility of a vertical transmission pathway wherein infected queens can pass virus through their eggs to their offspring.     

中蜂与意蜂无王群培育非自身蜂种蜂子及改造王台特性

以中华蜜蜂和意大利蜜蜂为试验材料,通过组织相应无王群,把一蜂种无王群中的子脾加入到另一蜂种无王群中,进行中蜂和意蜂无王群培育非自身蜂种蜂子和改造王台特性研究,当未培育出蜂王时,分别从原有王群中调入子脾进行第2、第3期试验。结果表明:中蜂无王群中的工蜂会清除引入意蜂子脾中的蜂卵,但随着引入意蜂子脾封盖子羽化出房的意蜂幼蜂数量增加而逐渐接受意蜂幼虫,蜂群未改造意蜂王台,只培育出中蜂蜂王;意蜂无王群中的工蜂会随着引入中蜂子脾中羽化出房的中蜂幼蜂数量增加而逐渐接受中蜂卵及幼虫,并从第2期试验开始会改造中蜂王台,但最终只培育出意蜂蜂王。     

温度对中华蜜蜂和意大利蜜蜂个体主要抗寒生理指标的影响

【目的】为了探究中华蜜蜂Apis cerana cerana和意大利蜜蜂Apis mellifera ligustica个体耐寒性差异的生理机制。【方法】本试验分别对20日龄的中华蜜蜂和意大利蜜蜂工蜂进行0、10、25℃持续4 h的温度处理,随后测定不同温度处理后两蜂种体内葡萄糖、甘油和氨基酸的含量。【结果】低温状态下中华蜜蜂体内葡萄糖含量显著降低,意大利蜜蜂在0℃组显著下降却在10℃组显著上升;中华蜜蜂体内甘油在低温环境下显著积累,意大利蜜蜂体内甘油水平在10℃时显著的上调却在0℃组无显著变化;中华蜜蜂体内17种氨基酸在各温度处理间显著变化,且多数的氨基酸含量表现出10℃组降低而0℃组积累的趋势,意大利蜜蜂体内只有15种氨基酸参与机体的抗寒机制,其中多数的氨基酸表现为低温环境下大量积累,仅有亮氨酸和精氨酸表现为低温时被转化消耗的现象。【结论】低温条件下可引起两蜂种体内葡萄糖、甘油和氨基酸含量的显著变化,初步推测蜜蜂体内可能存在"葡萄糖-甘油-氨基酸"的抗寒物质系统,但两蜂种在不同程度的低温处理后各生理指标的变化存在较大的差异,从一定程度上阐释两蜂种抗寒性的差异。本研究为蜜蜂个体抗寒生理机制的进一步研究提供基础数据,同时为蜜蜂抗寒新品系的选育提供理论依据。     

Comparison of learning and memory of Apis cerana and Apis mellifera

The honeybee is an excellent model organism for research on learning and memory among invertebrates. Learning and memory in honeybees has intrigued neuroscientists and entomologists in the last few decades, but attention has focused almost solely on the Western honeybee, Apis mellifera. In contrast, there have been few studies on learning and memory in the Eastern honeybee, Apis cerana. Here we report comparative behavioral data of color and grating learning and memory for A. cerana and A. mellifera in China, gathered using a Y-maze apparatus. We show for the first time that the learning and memory performance of A. cerana is significantly better on both color and grating patterns than that of A. mellifera. This study provides the first evidence of a learning and memory difference between A. cerana and A. mellifera under controlled conditions, and it is an important basis for the further study of the mechanism of learning and memory in honeybees.     

Molecular characterization and tissue localization of sensory neuron membrane protein from Chinese honey bee, Apis cerana cerana (Hymenoptera: Apidae)

Sensory neuron membrane protein (SNMP) is an olfactory receptor with photoaffinity analogs, capable of binding the pheromone membrane protein receptor deduced from receptor membrane protein with the pheromone–pheromone binding protein complex. However, this hypothesis has not yet been experimentally verified. In this experiment, the cDNA sequence encoding an open reading frame (ORF) of the SNMP gene AccSNMP1 (GenBank, KC012595) was cloned from Chinese honey bee, Apis cerana cerana Fabricius. Results from sequence analysis showed that this gene is 1,563 bp long, and that the ORF encodes 520 amino acids with a predicted molecular weight of 58.02 kDa, and has a theoretical isoelectric point of 5.83. Furthermore, there are two putative transmembrane domains. Multiple sequence alignment indicated that the AccSNMP1 gene from A. cerana cerana had different degrees of identity with the corresponding genes in nineteen other insects at the amino acid level. Phylogenetic analysis of the aligned sequences showed that A. cerana cerana is closely related to Apis mellifera Linnaeus and Bombus impatiens Cresson. Its distribution in tissues, as quantified using real-time RT-PCR, indicated that AccSNMP1 is highly expressed in the antennae and legs of A. cerana cerana, and there was a significant difference (p < 0.05) in gene expression between those tissues and tissues in the thorax, abdomen, snout, and head (not including antennae). Western blotting also confirmed the existence in the antennae of AccSNMP1 with an M _W of 58.0 kDa, which is the same as the expected value of 58.02 kDa. An immunohistochemistry study showed that AccSNMP1 is expressed in the trichoid sensilla of A. cerana cerana antenna. Therefore, the results of this study provide the basis for further studies of the function of SNMP from A. cerana cerana.     

中华蜜蜂与意大利蜜蜂蜂王体表信息素含量比较

【目的】分析中华蜜蜂Apis cerana cerana与意大利蜜蜂Apis mellifera ligustica蜂王不同时期的体表信息素含量变化,探索两蜂种之间蜂王交尾干扰机理。【方法】本试验通过GC-MS检测并分析了中华蜜蜂蜂王和意大利蜜蜂蜂王刚出房、性成熟时期以及婚飞过程中体表信息素含量变化。【结果】研究结果表明:中华蜜蜂性成熟蜂王在飞行过程中,其体表9-ODA含量显著高于刚出房蜂王,9-HDA显著高于刚出房蜂王和性成熟蜂王;意大利蜜蜂飞行蜂王在9-ODA含量也显著高于刚出房蜂王。另外,意大利蜜蜂性成熟期蜂王在9-ODA、9-HDA、10-HDA含量显著高于中华蜜蜂蜂王,而两蜂种蜂王体表信息素在婚飞时期差异不显著。【结论】同种蜂王不同发育时期,其体表信息素含量存在差异;中华蜜蜂蜂王与意大利蜂王在婚飞过程中,其体表信息素差异不显著,但部分体表信息素在性成熟而未进行婚飞时差异显著。     

Apis iridescent virus and "clustering disease" of Apis cerana

NPV of Spodoptera littoralis was completely inactivated in vitro following 10 min of exposure to a temperature higher than 90°C, but survived 3 weeks at ?20°C. At pH 12, some 75% of the infectivity was lost. Measurable proteolysis in vitro of the polyhedral protein by a larval midgut extract could be obtained only when the pH of the reaction mixture was raised to an unnatural level of 10.5, the natural pH of the midgut content being 8.5 or 9.5 according to different authors. The plant growth retardant Phosfon synergized mortality caused by the NPV. The virus could be cross-transmitted to two congeneric species of Spodoptera (S. exigua and S. litura), but could not infect any of four tested species belonging to other genera of the Moctuid family.     

Uncovering the novel characteristics of Asian honey bee,Apis cerana,by whole genome sequencing

Background

The honey bee is an important model system for increasing understanding of molecular and neural mechanisms underlying social behaviors relevant to the agricultural industry and basic science. The western honey bee, Apis mellifera, has served as a model species, and its genome sequence has been published. In contrast, the genome of the Asian honey bee, Apis cerana, has not yet been sequenced. A. cerana has been raised in Asian countries for thousands of years and has brought considerable economic benefits to the apicultural industry. A cerana has divergent biological traits compared to A. mellifera and it has played a key role in maintaining biodiversity in eastern and southern Asia. Here we report the first whole genome sequence of A. cerana.

Results

Using de novo assembly methods, we produced a 238 Mbp draft of the A. cerana genome and generated 10,651 genes. A.cerana-specific genes were analyzed to better understand the novel characteristics of this honey bee species. Seventy-two percent of the A. cerana-specific genes had more than one GO term, and 1,696 enzymes were categorized into 125 pathways. Genes involved in chemoreception and immunity were carefully identified and compared to those from other sequenced insect models. These included 10 gustatory receptors, 119 odorant receptors, 10 ionotropic receptors, and 160 immune-related genes.

Conclusions

This first report of the whole genome sequence of A. cerana provides resources for comparative sociogenomics, especially in the field of social insect communication. These important tools will contribute to a better understanding of the complex behaviors and natural biology of the Asian honey bee and to anticipate its future evolutionary trajectory.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-16-1) contains supplementary material, which is available to authorized users.     

中华蜜蜂工蜂嗅叶的胚后发育

通过形态解剖、免疫组织化学、原位细胞凋亡检测技术,对中华蜜蜂工蜂嗅叶的胚后发育过程进行了系统的比较研究.结果表明:(1)中华蜜蜂工蜂的嗅叶由神经纤维网区和外围的中脑神经细胞体层组成,在幼虫早期,神经纤维网的腹外侧和背外侧区内有几个大型的成神经细胞,呈现出典型的不对称分裂模式;在随后的发育过程中,神经纤维网的体积逐渐增加,神经细胞逐渐分散,细胞层变薄;分裂细胞主要集中在背外侧细胞区;在3龄幼虫期 ,触角嗅觉神经元的传入纤维开始进入嗅叶;在蛹发育的第3天,神经纤维网内的神经纤维球开始出现;(2)嗅叶发育过程中的细胞凋亡发生较少,增殖细胞和凋亡细胞数量的相对比率基本恒定;细胞凋亡的高峰期出现在幼虫末期;蛹发育的第4天左右,凋亡细胞完全消失 ;(3)中华蜜蜂的嗅叶存在雌雄异形现象[动物学报 54(3):546-554,2008].     

Detection of honey bee (Apis mellifera) viruses with an oligonucleotide microarray

In recent years, declines in honey bee (Apis mellifera L.) colonies have been observed to varying degrees worldwide with the worst losses in the USA being termed Colony Collapse Disorder (CCD). Pathogen load and the prevalence of honey bee viruses have been implicated in these losses and many diseased hives have multiple viruses present. We have designed and tested an oligonucleotide microarray which enables the simultaneous detection of nine honey bee viruses: Acute bee paralysis virus, Black queen cell virus, Chronic bee paralysis virus, Deformed wing virus, Kashmir bee virus, Sacbrood virus, Israel acute paralysis virus, Varroa destructor virus 1 and Slow paralysis virus. The microarray can be used to robustly diagnose nine viruses in one test.