Bacterial community structures in honeybee intestines and their response to two insecticidal proteins

In this study, the effects of the Bt-toxin Cry1Ab and a soybean trypsin inhibitor (SBTI) on intestinal bacterial communities of adult honeybees (Apis mellifera) were investigated. It was hypothesized that changes in intestinal bacterial communities of honeybees may represent a sensitive indicator for altered intestinal physiology. Honeybees were fed in a laboratory set-up with maize pollen from the Bt-transgenic cultivar MON810 or from the non-transgenic near isoline. Purified Cry1Ab (0.0014% w/v) and SBTI (0.1% or 1% w/v) represented supplementary treatments. For comparison, free-flying honeybees from two locations in Switzerland were analysed. PCR-amplification of bacterial 16S rRNA gene fragments and terminal restriction fragment length polymorphism analyses revealed a total of 17 distinct terminal restriction fragments (T-RFs), which were highly consistent between laboratory-reared and free-flying honeybees. The T-RFs were affiliated to Alpha-, Beta-, and Gammaproteobacteria, to Firmicutes, and to Bacteriodetes. Neither Bt-maize pollen nor high concentrations of Cry1Ab significantly affected bacterial communities in honeybee intestines. Only the high concentration of SBTI significantly reduced the number of T-RFs detected in honeybee midguts, a concentration that also increases bee mortality. Therefore, total bacterial community structures may not be a sensitive indicator for providing evidence for the impact of insecticidal proteins on honeybees at sublethal levels.     

Variation in Honey Bee Gut Microbial Diversity Affected by Ontogenetic Stage,Age and Geographic Location

Social honey bees, Apis mellifera, host a set of distinct microbiota, which is similar across the continents and various honey bee species. Some of these bacteria, such as lactobacilli, have been linked to immunity and defence against pathogens. Pathogen defence is crucial, particularly in larval stages, as many pathogens affect the brood. However, information on larval microbiota is conflicting.Seven developmental stages and drones were sampled from 3 colonies at each of the 4 geographic locations of A. mellifera carnica, and the samples were maintained separately for analysis. We analysed the variation and abundance of important bacterial groups and taxa in the collected bees.Major bacterial groups were evaluated over the entire life of honey bee individuals, where digestive tracts of same aged bees were sampled in the course of time. The results showed that the microbial tract of 6-day-old 5th instar larvae were nearly equally rich in total microbial counts per total digestive tract weight as foraging bees, showing a high percentage of various lactobacilli (Firmicutes) and Gilliamella apicola (Gammaproteobacteria 1). However, during pupation, microbial counts were significantly reduced but recovered quickly by 6 days post-emergence. Between emergence and day 6, imago reached the highest counts of Firmicutes and Gammaproteobacteria, which then gradually declined with bee age. Redundancy analysis conducted using denaturing gradient gel electrophoresis identified bacterial species that were characteristic of each developmental stage.The results suggest that 3-day 4th instar larvae contain low microbial counts that increase 2-fold by day 6 and then decrease during pupation. Microbial succession of the imago begins soon after emergence. We found that bacterial counts do not show only yearly cycles within a colony, but vary on the individual level. Sampling and pooling adult bees or 6th day larvae may lead to high errors and variability, as both of these stages may be undergoing dynamic succession.     

意大利蜜蜂(Apis mellifera ligustica)与中华蜜蜂(Apis cerana ceraca)的生态位比较

为了明确中华蜜蜂和意大利蜜蜂在皖南山区生态适应性,研究两种蜜蜂的食物生态位、时间生态位和空间生态位及其差异,结果是中蜂与意蜂食物(蜜源植物)资源生态位宽度分别是 0.923、0.765,中蜂对蜜源植物采集喜好性差异小,而意蜂差异大,中蜂对意蜂生态位重迭为0.160,中蜂对意蜂生态位相似性为0.755;油菜花期,中蜂与意蜂时间资源生态位宽度分别是0.879、0.801,枇杷花期,分别是0.760、0.677,中蜂与意蜂的空间资源生态位宽度分别是0.797、0.670.中蜂3种生态位宽度均大于意蜂,中蜂三维生态位值是意蜂的1.61倍和1.57倍.表明中蜂在皖南山区生态适应性比意蜂强.     

Natural selection of Varroa jacobsoni explains the different reproductive strategies in colonies of Apis cerana and Apis mellifera

In colonies of European Apis mellifera, Varroa jacobsoni reproduces both in drone and in worker cells. In colonies of its original Asian host, Apis cerana, the mites invade both drone and worker brood cells, but reproduce only in drone cells. Absence of reproduction in worker cells is probably crucial for the tolerance of A. cerana towards V. jacobsoni because it implies that the mite population can only grow during periods in which drones are reared. To test if non-reproduction of V. jacobsoni in worker brood cells of A. cerana is due to a trait of the mites or of the honey-bee species, mites from bees in A. mellifera colonies were artificially introduced into A. cerana worker brood cells and vice versa. Approximately 80% of the mites from A. mellifera colonies reproduced in naturally infested worker cells as well as when introduced into worker cells of A. mellifera and A. cerana. Conversely, only 10% of the mites from A. cerana colonies reproduced, both in naturally infested worker cells of A. cerana and when introduced into worker cells of A. mellifera. Hence, absence of reproduction in worker cells is due to a trait of the mites. Additional experiments showed that A. cerana bees removed 84% of the worker brood that was artificially infested with mites from A. mellifera colonies. Brood removal started 2 days after artificial infestation, which suggests that the bees responded to behaviour of the mites. Since removal behaviour of the bees will have a large impact on fitness of the mites, it probably plays an important role in selection for differential reproductive strategies. Our findings have large implications for selection programmes to breed less-susceptible bee strains. If differences in non-reproduction are mite specific, we should not only look for non-reproduction as such, but for colonies in which non-reproduction in worker cells is selected. Hence, in selection programmes fitness of mites that reproduce in both drone and worker cells should be compared to fitness of mites that reproduce only in drone cells. © Rapid Science Ltd. 1998     

Racial Differences in Division of Labor in Colonies of the Honey Bee (Apis mellifera)

We measured the age at onset of foraging in colonies derived from three races of European honey bees, Apis mellifera mellifera, Apis mellifera caucasica and Apis mellifera ligustica , using a cross-fostering design that involved six unrelated colonies of each race. There was a significant effect of the race of the introduced bees on the age at onset of foraging: cohorts of A. m. ligustica bees showed the earliest onset, regardless of the race of the colony they were introduced to. There also was a significant effect of the race of the host colony: cohorts of bees introduced into mellifera colonies showed the earliest onset of foraging, regardless of the race of the bees introduced. Significant inter-trial differences also were detected, primarily because of a later onset of foraging in trials conducted during the autumn (September–October). These results demonstrate differences among European races of honey bees in one important component of colony division of labor. They also provide a starting point for analyses of the evolution of division of labor under different ecological conditions.     

中、西蜂间自然交尾干扰问题的观察

中西蜂蜂王性引诱信息素对中、西蜂雄蜂具有交互引诱交尾作用 ,且处女王婚飞和雄蜂出游时间一致。在中、西蜂 (主要指意蜂 )混养区 ,空中交尾竞争处于劣势的中蜂 ,受到西蜂严重干扰而繁殖受阻 ,是中蜂在平原等地区日益缩减多因素中的一个重要原因。蜂种间“自然交尾干扰”论点的提出 ,为保护中蜂资源、发展中蜂生产提供了科学依据。     

Diversity and phylotype consistency of bacteria in the guts of three bee species (Apoidea) at an oilseed rape field

The gut of insects may harbour one of the largest reservoirs of a yet unexplored microbial diversity. To understand how specific insects select for their own bacterial communities, the structural diversity and variability of bacteria found in the gut of different bee species was analysed. For three successive years, adults and larvae of Apis mellifera ssp. carnica (honey bee), and Bombus terrestris (bumble bee), as well as larvae of Osmia bicornis (red mason bee) were collected at a flowering oilseed rape field. Total DNA was extracted from gut material and the bacterial diversity was analysed, independent of cultivation, by genetic profiling with single-strand conformation polymorphism (SSCP) of polymerase chain reaction (PCR)-amplified partial 16S rRNA genes. The SSCP profiles were specific for all bee species and for larvae and adults. Qualitative and quantitative differences were found in the bacterial community structure of larvae and adults of A. mellifera, but differences in B. terrestris were mainly quantitative. Sequencing of the PCR products revealed a dominance of Alpha-, Beta-, and Gammaproteobacteria, Bacteroidetes, and Firmicutes in all bee species. Single-strand conformation polymorphism profiles suggested a higher abundance and diversity of lactobacilli in adults of A. mellifera than in larvae. Further phylogenetic analyses indicated common bacterial phylotypes for all three bee species, e.g. those related to Simonsiella, Serratia, and Lactobacillus. Clades related to Delftia acidovorans, Pseudomonas aeruginosa or Lactobacillus intestinalis only contained sequences from larvae. Several of the bee-specific clusters also contained identical or highly similar sequences from bacteria detected in other A. mellifera subspecies from South Africa, suggesting the existence of cosmopolitan gut bacteria in bees.     

Nosema ceranae is a long-present and wide-spread microsporidian infection of the European honey bee (Apis mellifera) in the United States

Honey bee samples collected between 1995 and 2007 from 12 states were examined for the presence of Nosema infections. Our results showed that Nosema ceranae is a wide-spread infection of the European honey bee, Apis mellifera in the United States. The discovery of N. ceranae in bees collected a decade ago indicates that N. ceranae was transferred from its original host, Apis cerana to A. mellifera earlier than previously recognized. The spread of N. ceranae infection in A. mellifera warrants further epidemiological studies to identify conditions that resulted in such a widespread infection.     

皖南山区不同生态条件下中华蜜蜂形态特征差异性

为了分析皖南山区中华蜜蜂的生态适应性,对皖南山区5个不同采样点的15群中华蜜蜂450只工蜂的8个形态特征进行测定,对测定的性状特征数据进行数学分析和差异显著性检验,结果是,5个釆样点间的喙长、右前翅长、第三节背板宽、第三节背板颜色、第四节背板宽和第四节背板颜色差异显著。皖南山区中华蜜蜂的前翅长与宽均较大,第3+4背板长也较大,说明皖南中蜂飞翔能力强,采集能力强;背板长反映蜜囊的大小,蜜囊越大,贮存花蜜越多,蜂群进蜜就越快,从形态上说明皖南中蜂是较理想的蜜蜂种群。皖南中蜂种群内遗传变异丰富,种群形态性状的多态性、多型性及其生态地理变异式样,具有生态适应意义。     

Phylogenetic relationships of honey bees (Hymenoptera:Apinae:Apini) inferred from nuclear and mitochondrial DNA sequence data

Two different genomic regions (ND2 mitochondrial gene and EF1-alpha intron) were PCR amplified, cloned and sequenced for the ten known honey bee species collected within their natural range distribution. DNA sequences were analyzed using parsimony, distance and maximum likelihood methods to investigate phylogenetic relationships within Apis. The phylogenetic analyses strongly supported the basic topology recoverable from morphometric analysis, grouping the honey bees into three major clusters: giant bees (A. dorsata, A. binghami, and A. laboriosa), dwarf bees (A. andreniformis and A. florea), and cavity-nesting bees (A. mellifera, A. cerana, A. koschevnikovi, A. nuluensis, and A. nigrocincta). However, the clade of Asian cavity-nesting bees included paraphyletic taxa. Exemplars of Apis cerana collected from divergent portions of its range were less related to each other than were sympatric A. cerana, A. nuluensis, and A. nigrocincta taxa. Nucleotide sequence divergence between allopatrically distributed western (A. mellifera) and eastern (A. cerana, A. koschevnikovi, A. nigrocincta, and A. nuluensis) cavity-nesting species, around 18% for the mitochondrial gene and 10-15% for the nuclear intron, suggested an earlier divergence for these groups than previously estimated from morphometric and behavioral studies. This latter finding neccessitates reevaluation of the hypothesized origin of extant European, African, and west Asian Apis mellifera. Sequence divergence between A. laboriosa and A. dorsata was consistent with behavioral data and supports the species status of A. laboriosa.     

从有意引入到外来入侵--以意大利蜂Apis mellifera L.为例

原产于欧洲、非洲等地的意大利蜜蜂几个世纪以来被人类频繁地有意引入到其它国家和地区,然而其在带来经济利益的同时,意大利蜂凭借其在诸多方面的竞争优势对当地物种和生态系统产生不利影响。本文主要阐述了意大利蜜蜂引入后对我国本地种——中华蜜蜂造成的负面效应：①中华蜜蜂种群数量急剧下降,甚至在某些地区已到了灭绝的边缘;②影响了以中华蜜蜂作为主要传粉媒介的早花乔木、灌木和草本植物的正常繁育,从而降低了物种多样性,破坏了群落结构及其稳定性。     

基于线粒体和核基因序列的蜜蜂属系统发育分析

文章测定了中国分布的蜜蜂属(Apis)5种蜜蜂22个样本的线粒体基因ND2、CO2、16S rRNA以及核基因ITPR的序列,对序列的碱基组成和蜜蜂种间的遗传距离进行了分析。结合下载的蜜蜂属其他4个种的相关序列,采用最大简约法、邻接法和最大似然法重建了蜜蜂属系统发育关系。系统发育分析结果支持蜜蜂属划分为3个类群,即小蜜蜂类群(包括小蜜蜂和黑小蜜蜂)、大蜜蜂类群(包括大蜜蜂和黑大蜜蜂)和穴居蜜蜂类群(西方蜜蜂、东方蜜蜂、沙巴蜂、苏拉威西蜂、绿努蜂),且小蜜蜂类群较早分化。结果还显示,我国海南岛的大蜜蜂和大陆的大蜜蜂之间可能存在较大的遗传分歧。     

Open-air-nesting honey bees Apis dorsata and Apis laboriosa differ from the cavity-nesting Apis mellifera and Apis cerana in brood hygiene behaviour

The cavity-nesting Apis mellifera and Apis cerana bees detect, uncap, and remove diseased brood. The hygiene behaviour of open-air-nesting bees Apis dorsata and Apis laboriosa was investigated in India and Nepal. Sealed A. dorsata pupae were pin-killed or deep-frozen. The workers removed 73 or 37% of damaged pin-killed pupae depending on the diameter of the pins, and only 7% of the frozen undamaged pupae. Migrating A. dorsata and A. laboriosa left unopened the sealed brood in deserted combs. Thus, A. dorsata and A. laboriosa do not open undamaged cells with dead brood. This behaviour is a more efficient mechanism in preventing the spread of diseases and parasitic mites than uncapping and removing dead pupae by A. mellifera and A. cerana. It may be beneficial for migrating A. dorsata and A. laboriosa to temporarily disuse part of the comb cells in exchange for arresting the mites there and thus reducing the increase of their population.     

Distinctive gut microbiota of honey bees assessed using deep sampling from individual worker bees

Surveys of 16S rDNA sequences from the honey bee, Apis mellifera, have revealed the presence of eight distinctive bacterial phylotypes in intestinal tracts of adult worker bees. Because previous studies have been limited to relatively few sequences from samples pooled from multiple hosts, the extent of variation in this microbiota among individuals within and between colonies and locations has been unclear. We surveyed the gut microbiota of 40 individual workers from two sites, Arizona and Maryland USA, sampling four colonies per site. Universal primers were used to amplify regions of 16S ribosomal RNA genes, and amplicons were sequenced using 454 pyrotag methods, enabling analysis of about 330,000 bacterial reads. Over 99% of these sequences belonged to clusters for which the first blastn hits in GenBank were members of the known bee phylotypes. Four phylotypes, one within Gammaproteobacteria (corresponding to "Candidatus Gilliamella apicola") one within Betaproteobacteria ("Candidatus Snodgrassella alvi"), and two within Lactobacillus, were present in every bee, though their frequencies varied. The same typical bacterial phylotypes were present in all colonies and at both sites. Community profiles differed significantly among colonies and between sites, mostly due to the presence in some Arizona colonies of two species of Enterobacteriaceae not retrieved previously from bees. Analysis of Sanger sequences of rRNA of the Snodgrassella and Gilliamella phylotypes revealed that single bees contain numerous distinct strains of each phylotype. Strains showed some differentiation between localities, especially for the Snodgrassella phylotype.     

Investigation of gut microbial communities associated with indigenous honey bee (Apis mellifera jemenitica) from two different eco-regions of Saudi Arabia

The microbial communities associated with the alimentary tract of honey bees are very important as they help with food digestion, provide essential nutrients, protect the host from pathogens, detoxify harmful molecules, and increase host immunity. In this study, the structural diversity of the gut microbial communities of native honey bees, Apis mellifera jemenitica from two different geographical regions (Riyadh and Al-Baha) of Saudi Arabia was analyzed by culture-dependent methods and 16S ribosomal RNA (rRNA) gene sequencing. In this study, 100 bacterial isolates were cultivated and phylogenetic analyses grouped them into three phyla: Proteobacteria, Firmicutes, and Actinobacteria. Bacteria in the phylum Proteobacteria were the most dominant (17 species), followed by Firmicutes (13 species) and Actinobacteria (4 species). Some of the identified bacteria (Citrobacter sp., Providencia vermicola, Exiguobacterium acetylicum, and Planomicrobium okeanokoites) were reported for the first time in the genus Apis, while others identified bacteria belonged to the genera Proteus, Enterobacter, Bacillus, Morganella, Lactobacillus, and Fructobacillus. To the best of our knowledge, this is the first study on the gut microbiota of the local honey bees in Saudi Arabia.     

Morphometrics and adaptatives aspects in africanized honeybees (Apis mellifera)

The introduction of the African bees (Apis mellifera scutelata) in Brazil and their expansion in the American Continent created the opportunity to study the process of species adaptation in a new environment. In that process, within intra-specific variability, normalising selection can favour individuals that present a better adaptative morphology and they will constitute the most frequent type found in the population. To test that hypothesis morphometric analyses in samples of colonies of africanized bees and in samples of the populations were performed. The development of the colonies was also evaluated in terms of the amount of their brood, honey and pollen. Analysis of the data indicates that more developed colonies are formed by individuals closer to the population average with concerning morphological traits.     

Structural and functional differences in the antennal olfactory system of worker honey bees of Apis mellifera and Apis cerana

Olfactory cues are important sensory modalities on individual discrimination, perception, and efficient orientation to food sources in most insects. In honey bees, which are well known as eusocial insects, olfactory cues are mainly used to maintain a colony. Although much research has been reported on olfactory systems in honey bee olfaction, little is known about the differences between two major honey bee species, the European honey bee Apis mellifera and the Asian honey bee Apis cerana. In order to understand the differences of olfactory characteristics in the two species, we compared the distribution of sensory hairs on the antennae and antennal olfactory responses, using electron microscopy, electrophysiological recording and molecular expression level of odorant receptors. Our present study demonstrated that the antennae of A. cerana have more olfactory sensilla than A. mellifera, responding more strongly to various floral volatile compounds. At the molecular level, olfactory co-receptor (Orco), which makes heterodimers with other conventional olfactory receptors, is more abundantly expressed in the antenna of A. cerana than of A. mellifera. These findings extend our understanding of the olfactory systems and behavioral responses to various ecological and biological signals in two closely related honey bee species.     

The accuracy of morphometric characteristic analysis depends on the type of the assessed traits of honey bees (Apis cerana F. and Apis mellifera L.)

Conservation of honey bees as pollinators and honey producers requires their assessment using convenient general methods. One of the most commonly used methods involves assessment of morphometric characteristics using random traits. The purpose of the study was to determine a set of morphometric characteristics for identifying the local (R, H: Apis cerana koreana) and adapted (A, C, F: Apis melifera ligustica; D: Apis mellifera carpathica; V: Apis mellifera) lines of honey bees bred in Korea. We found traits with significant differences (p < 0.001), high discriminant coefficient (>100), and range of variation (<50 %). The analysis demonstrated that the widths of the abdomen, forewing, and head, and the lengths of the antenna, body, proboscis, tergites 4 and 3, and forewing can be used as the nine main morphological characteristics for distinguishing the bred lines of honey bees. Furthermore, the last two traits and the length of the head can be used for distinguishing the adapted subspecies A. m. ligustica and A. m. carpathica. Our observations can be used for improving the morphometric method for the conservation of honey bees.     

High genetic variability of Nosema ceranae populations in Apis mellifera from East Asia compared to central Asia and the Americas

Biological Invasions - Nosema ceranae is believed to have been originally a parasite of the Asian honey bee, Apis cerana, in East Asia that later infected the Western honey bee, Apis mellifera,...     

基于线粒体Cyt b基因部分序列的中国东方蜜蜂不同地理种群的系统发育

从线粒体DNA水平上对中国境内东方蜜蜂不同地理种群的系统发育进行研究,为保护和合理利用这一宝贵的蜂种资源提供理论基础。对我国10个省市的21群东方蜜蜂mtDNA Cyt b基因片段进行了扩增、测序,并以意大利蜂、卡尼鄂拉蜂、印尼蜂作为外群进行序列分析。采用Minimum-Spanning Network方法构建系统进化树。结果显示：扩增片段长度429bp;在得到的21条同源序列中,共检测出13个变异位点,其中转换数为9,颠换数为4,并且变异位点大都发生在密码子的第三位,无碱基插入或缺失;聚类分析结果显示西方蜜蜂、印尼蜂和东方蜜蜂各为独立的蜂种;在中国东方蜜蜂群体中,吉林、海南和云南的东方蜜蜂各为一个独立的类群,其它地区的东方蜜蜂为一个类群[动物学报54（6）：1005-1013,2008]。