Molecular and phylogenetic characterization of honey bee viruses,Nosema microsporidia,protozoan parasites,and parasitic mites in China

China has the largest number of managed honey bee colonies, which produce the highest quantity of honey and royal jelly in the world; however, the presence of honey bee pathogens and parasites has never been rigorously identified in Chinese apiaries. We thus conducted a molecular survey of honey bee RNA viruses, Nosema microsporidia, protozoan parasites, and tracheal mites associated with nonnative Apis mellifera ligustica and native Apis cerana cerana colonies in China. We found the presence of black queen cell virus (BQCV), chronic bee paralysis virus (CBPV), deformed wing virus (DWV), Israeli acute paralysis virus (IAPV), and sacbrood virus (SBV), but not that of acute bee paralysis virus (ABPV) or Kashmir bee virus (KBV). DWV was the most prevalent in the tested samples. Phylogenies of Chinese viral isolates demonstrated that genetically heterogeneous populations of BQCV, CBPV, DWV, and A. cerana‐infecting SBV, and relatively homogenous populations of IAPV and A. meliifera‐infecting new strain of SBV with single origins, are spread in Chinese apiaries. Similar to previous observations in many countries, Nosema ceranae, but not Nosema apis, was prevalent in the tested samples. Crithidia mellificae, but not Apicystis bombi was found in five samples, including one A. c. cerana colony, demonstrating that C. mellificae is capable of infecting multiple honey bee species. Based on kinetoplast‐encoded cytochrome b sequences, the C. mellificae isolate from A. c. cerana represents a novel haplotype with 19 nucleotide differences from the Chinese and Japanese isolates from A. m. ligustica. This suggests that A. c. cerana is the native host for this specific haplotype. The tracheal mite, Acarapis woodi, was detected in one A. m. ligustica colony. Our results demonstrate that honey bee RNA viruses, N. ceranae, C. mellificae, and tracheal mites are present in Chinese apiaries, and some might be originated from native Asian honey bees.     

蜜蜂及熊蜂微孢子虫rRNA基因研究

核糖体RNA(rRNA)是一种理想的进化计时器,已广泛地应用于研究微生物的系统发育和进化关系。蜜蜂和熊蜂都被微孢子虫感染,关于蜜蜂和熊蜂微孢子虫rRNA基因研究主要有rRNA基因序列的测定分析、二级结构的研究和用rRNA基因检测诊断微孢子虫等。这些对蜜蜂和熊蜂微孢子虫系统发育的研究和微孢子虫的防治等具有重要的推动作用。     

Brain melatonin content and polyethism in adult workers of Apis mellifera and Apis cerana (Hym., Apidae)

Abstract:  Radioimmunoassay showed that melatonin, N -acetyl-5-methoxytryptamine, was synthesized in the brain of workers of Apis mellifera ligustica Spinola, and related to the polyethism of the workers. Statistically significant differences were found in melatonin levels between workers at different ages and between workers performing different tasks. The level in foragers was higher than that in young nurses. The highest melatonin level was found in 21-day-old adult workers (213.15 pg per worker head) and the lowest level in 1-day-old adult workers (6.08 pg per worker head); and the highest level in pollen-collecting workers (404.13 pg per worker head) and the lowest in nurse workers (13.10 pg per worker head). Apis cerana cerana Fabricius showed a similar pattern, but the nectar foragers had the highest melatonin levels. The melatonin level also showed a daily rhythm. In 30-day-old adult workers there were two peaks, at 7:00 and 19:00 hours, and three troughs at 15:00, 21:00 and 1:00 hours respectively.     

Floral resources collected by four native bees species in southern Mexico

Abstract

Melissopalynological analysis of 14 honey samples of Tetragonisca angustula (two), Scaptotrigona mexicana (six), Melipona beecheii (three) and Melipona solani (three) from Soconusco region in Chiapas, Mexico was performed. A total of 79 taxa were identified, being Asteraceae, Euphorbiaceae, Fabaceae, Melostomataceae and Rubiaceae the predominant pollen types. Melissopalynological analysis reported a monofloral composition for Melipona beecheii honeys, where the most predominant pollen type was Fabaceae (54.2%). T. angustula, Scaptotrigona mexicana and Melipona solani honey samples were multifloral and predominant pollen types were Asteraceae, Euphorbiaceae, Fabaceae, Melastomataceae and Solanaceae. The most frequent polliniferous pollen type were Asteraceae, Bernardia interrupta, Euphorbia heterophyla and Miconia. Shannon–Wiener diversity index (H′) for Melipona beecheii 2016 (2.21) and Melipona solani 2017 (1.64) were lower meanwhile honeys of Melipona beecheii 2017, S. mexicana 2017, Melipona solani 2016 and T. angustula 2017 were considered as fairly diverse, because they visited a wider range of plant species, by preferring shrub and wild strata such as: Asteraceae, Euphorbia, Euphorbiaceae, Miconia and Mimosa. Our results showed that the foraging behaviour of these bee species is polylectic, as no taxa was found to be more than 70% of the counted grains. Since the study of the collection of meliponine flower resources in the southern region of Mexico have not been studied yet, it is not possible to analyse the foraging behaviour, the use of resources and the management of these species in this region.     

Strikingly high levels of heterozygosity despite 20 years of inbreeding in a clonal honey bee

Inbreeding (the mating between closely related individuals) often has detrimental effects that are associated with loss of heterozygosity at overdominant loci, and the expression of deleterious recessive alleles. However, determining which loci are detrimental when homozygous, and the extent of their phenotypic effects, remains poorly understood. Here, we utilize a unique inbred population of clonal (thelytokous) honey bees, Apis mellifera capensis, to determine which loci reduce individual fitness when homozygous. This asexual population arose from a single worker ancestor approximately 20 years ago and has persisted for at least 100 generations. Thelytokous parthenogenesis results in a 1/3 of loss of heterozygosity with each generation. Yet, this population retains heterozygosity throughout its genome due to selection against homozygotes. Deep sequencing of one bee from each of the three known sub‐lineages of the population revealed that 3,766 of 10,884 genes (34%) have retained heterozygosity across all sub‐lineages, suggesting that these genes have heterozygote advantage. The maintenance of heterozygosity in the same genes and genomic regions in all three sub‐lineages suggests that nearly every chromosome carries genes that show sufficient heterozygote advantage to be selectively detrimental when homozygous.     

Regulation of nectar collection in relation to honey storage levels by honey bees, Apis mellifera

Honey bees collect distinct nutrient sources in the form ofnectar (energy) and pollen (nitrogen). We investigated the effectof varying energy stores on nectar and pollen foraging. We foundno significant changes in nectar foraging in response to changesin honey storage levels within colonies. Individual foragersdid not vary activity rates or nectar load sizes in responseto changes in honey stores, and colonies did not increase nectarintake rates when honey stores within the hive were decreased.This result contrasts with pollen foraging behavior, which isextremely sensitive to colony state. Our data show that individualforaging decisions during nectar collection and colony regulationof nectar intake are distincdy different from pollen foraging.The behavior of honey bees illustrates that foraging strategy(and therefore foraging models) can incorporate multiple currencies,including both energy and protein intake.[Behav Ecol 7: 286–291(1996)]     

生物行为变化研究的新模式——工蜂

蜜蜂群体由1只蜂王、几百只雄蜂和数千只工蜂组成,工蜂数量巨大,除蜂王和雄蜂共同完成生殖任务,巢内外活动均由工蜂完成,其行为呈现多样性。工蜂发育经过卵、幼虫、蛹、成虫4个阶段,其活动范围由温度、湿度相对稳定的巢内环境发展到复杂的巢外自然环境。随发育阶段和生活环境的变化,工蜂的生理、行为等方面也发生相应变化,这种变化为综合研究生物行为的分子机理提供了可能。又因工蜂数量多、体积较大易于观察、标记、饲养管理简单,而且目前对其形态、发育、生理、分子生物学、神经、社会生态等各方面的研究比较充分,所以工蜂成为研究生物体行为变化、发育机理和个体与群体关系的理想模式生物。该文介绍工蜂活动由巢内发展到巢外时出现的一系列变化以及部分变化的机制,主要包括行为的变化、激素的分泌、代谢活动、飞行能力、神经系统等。     

蜜蜂寿命可塑性研究进展

蜜蜂是一种完全社会化的昆虫,它们在寿命方面表现出了显著的级型差异.蜂王的平均寿命是1～2年,而工蜂在生产季节平均寿命是30～40 d,越冬季节平均寿命是90～200d,显然蜜蜂寿命有可塑性.这种可塑性是由环境因素控制,因为蜂王和工蜂的遗传基础是一致的.另外工蜂任务可以发生逆转,这意味着老化的逆转.本文综述了近年来一些老...     

Floral bagging differentially affects handling behaviours and single-visit pollen deposition by honey bees and native bees

1. Measurements of pollinator performance are crucial to pollination studies, enabling researchers to quantify the relative value of different pollinator species to plant reproduction. One of the most widely employed measures of pollinator performance is single-visit pollen deposition, the number of conspecific pollen grains deposited to a stigma after one pollinator visit. To ensure a pollen-free stigma, experimenters must first bag flowers before exposing them to a pollinator. 2. Bagging flowers, however, may unintentionally manipulate floral characteristics to which pollinators respond. In this study, we quantified the effect of bagging on nectar volume in watermelon (Citrullus lanatus) flowers, and how this affects pollinator performance and behaviour. 3. Experimental bagging resulted in roughly 30-fold increases in nectar volume relative to unmanipulated, open-pollinated field flowers after only a few hours. Honey bees, but not native bees, consistently displayed elevated handling times and single-visit pollen deposition on unmanipulated bagged flowers relative to those from which we removed nectar to mimic volumes in open-pollinated flowers. 4. Furthermore, we identify specific bee foraging behaviours during a floral visit that account for differences in pollen deposition, and how these differ between honey bees and native bees. 5. Our findings suggest that experimental bagging of flowers, without accounting for artificially accumulated nectar, can lead to biased estimates of pollinator performance in pollinator taxa that respond strongly to nectar volume. We advise that pollination studies be attentive to nectar secretion dynamics in their focal plant species to ensure unbiased estimates of pollinator performance across multiple pollinator species.     

A Causal Analysis of Observed Declines in Managed Honey Bees (Apis mellifera)

The European honey bee (Apis mellifera) is a highly valuable, semi-free-ranging managed agricultural species. While the number of managed hives has been increasing, declines in overwinter survival, and the onset of colony collapse disorder in 2006, precipitated a large amount of research on bees’ health in an effort to isolate the causative factors. A workshop was convened during which bee experts were introduced to a formal causal analysis approach to compare 39 candidate causes against specified criteria to evaluate their relationship to the reduced overwinter survivability observed since 2006 of commercial bees used in the California almond industry. Candidate causes were categorized as probable, possible, or unlikely; several candidate causes were categorized as indeterminate due to lack of information. Due to time limitations, a full causal analysis was not completed at the workshop. In this article, examples are provided to illustrate the process and provide preliminary findings, using three candidate causes. Varroa mites plus viruses were judged to be a “probable cause” of the reduced survival, while nutrient deficiency was judged to be a “possible cause.” Neonicotinoid pesticides were judged to be “unlikely” as the sole cause of this reduced survival, although they could possibly be a contributing factor.     

Survey of insect visitation of ornamental flowers in Southover Grange garden,Lewes, UK

Ornamental flowers commonly grown in urban gardens and parks can be of value to flower‐visiting insects. However, there is huge variation in the number of insects attracted among plant varieties. In this study, we quantified the insect attractiveness of 79 varieties in full bloom being grown in a public urban garden that is popular due to its beautiful flowers and other attractions. The results showed very clearly that most varieties (77%, n = 61) were either poorly attractive or completely unattractive to insect flower visitors. Several varieties (19%, n = 15) were moderately attractive, but very few (4%, n = 3) were highly attractive. Closer examination of Dahlia varieties showed that “open” flowered forms were approximately 20 times more attractive than “closed” flowered forms. These results strongly suggest that there is a great potential for making urban parks and gardens considerably more bee‐ and insect‐friendly by selecting appropriate varieties.     

Detection of aerosolized bacterial spores (Bacillus atrophaeus) using free-flying honey bees (Hymenoptera: Apidae) as collectors

An aerosol cloud of Bacillus atrophaeus (previously B. subtilis variety niger) spores, an anthrax surrogate, was created in a large 0.4 ha (1 ac), bee-containing, open-mesh tent. Bees from a B. atrophaeus uncontaminated hive flying through the cloud adsorbed the spores in statistically significant quantities. After removal of the B. atrophaeus contaminated hive and introduction of another B. atrophaeus uncontaminated hive, the bees again were monitored for the next few days for B. atrophaeus spores. B. atrophaeus spores accumulated on the bees bodies following their exposure to the residual B. atrophaeus contamination in the tent. The spore loads on the bees quickly returned to background levels after the hives were removed from the contaminated tent area. It may therefore be practical to use honey bee colonies to monitor foraging areas for disease-causing spores.     

Detection of aerosolized bacterial spores (<Emphasis Type="Italic">Bacillus atrophaeus</Emphasis>) using free-flying honey bees (Hymenoptera: Apidae) as collectors

An aerosol cloud of Bacillus atrophaeus (previously B. subtilis variety niger) spores, an anthrax surrogate, was created in a large 0.4 ha (1 ac), bee-containing, open-mesh tent. Bees from a B. atrophaeus uncontaminated hive flying through the cloud adsorbed the spores in statistically significant quantities. After removal of the B. atrophaeus contaminated hive and introduction of another B. atrophaeus uncontaminated hive, the bees again were monitored for the next few days for B. atrophaeus spores. B. atrophaeus spores accumulated on the bees bodies following their exposure to the residual B. atrophaeus contamination in the tent. The spore loads on the bees quickly returned to background levels after the hives were removed from the contaminated tent area. It may therefore be practical to use honey bee colonies to monitor foraging areas for disease-causing spores.     

Colonization history and population differentiation of the Honey Bees (Apis mellifera L.) in Puerto Rico

Honey bees (Apis mellifera L.) are the primary commercial pollinators across the world. The subspecies A. m. scutellata originated in Africa and was introduced to the Americas in 1956. For the last 60 years, it hybridized successfully with European subspecies, previous residents in the area. The result of this hybridization was called Africanized honey bee (AHB). AHB has spread since then, arriving to Puerto Rico (PR) in 1994. The honey bee population on the island acquired a mosaic of features from AHB or the European honey bee (EHB). AHB in Puerto Rico shows a major distinctive characteristic, docile behavior, and is called gentle Africanized honey bees (gAHB). We used 917 SNPs to examine the population structure, genetic differentiation, origin, and history of range expansion and colonization of gAHB in PR. We compared gAHB to populations that span the current distribution of A. mellifera worldwide. The gAHB population is shown to be a single population that differs genetically from the examined populations of AHB. Texas and PR groups are the closest genetically. Our results support the hypothesis that the Texas AHB population is the source of gAHB in Puerto Rico.     

城市园林对本土传粉者中华蜜蜂的保护作用

【目的】蜜蜂等传粉昆虫是生态系统的关键种,其种群下降趋势是目前全球关注的热点问题,探讨传粉昆虫现状和保护途径具有重要实践意义。【方法】对四川地区城市园林里中华蜜蜂Apis cerana cerana进行调查,记录其拜访行为、蜜粉源植物和营巢生境特征,并观察主要植物的其他访花昆虫。【结果】城市园林中有大量开花植物,花期持续时间长,是蜜蜂可利用的食物资源;中华蜜蜂在城市中广泛活动,所调查城市都观测到采集蜂进行有效采集,在冬季中华蜜蜂蜂群也能正常活动;城市中其他访花昆虫以鳞翅目和双翅目为主,蝴蝶等会竞争食物资源,行为分析认为蜜蜂具有竞争优势;部分人工绿地和半自然生境可成为中华蜜蜂的营巢地。【结论】城市园林可为中华蜜蜂提供适宜的生存环境,建议改善城市景观和园林管理以更好地发挥园林生境的保护作用。     

Pollen analysis of honeys of Melipona (Michmelia) seminigra merrillae and Melipona (Melikerria) interrupta (Hymenoptera: Apidae) bred in Central Amazon,Brazil

The pollen present in honey in colonies of Melipona seminigra merrillae and Melipona interrupta bred in Manaus was analysed. Between August and October 2012, honey samples were collected from the Sucupira meliponary, located in Manaus, Amazonas, Brazil. We identified a total of 70 pollen types belonging to 35 botanical families. In the samples from Melipona seminigra merrillae, the most represented pollen types were: Miconia-type (Melastomataceae) was the dominant pollen (DP) in August (51.19%), September (76.83%) and October (58.33%); Triplaris-type (Polygonaceae) was an accessory pollen (AP) in August (20.5%); and the remaining pollen types were classified as isolated pollen (IP), with Talisia macrophylla (Sapindaceae) exhibiting the highest percentage in August (12.92%). For Melipona interrupta, the most frequent pollen types were as follows: Miconia-type was the DP in August (59.33%) and October (61.33%) and an AP in September (37.5%); and Triplaris-type was an AP in August (35.83%), September (34.16%) and October (23.33%). The diversity of pollen types was not significantly different between the bees in the months evaluated. However, there was a large significant niche overlap in the months studied, August (O_ik = 0.95), October (O_ik = 0.89) and September (O_ik = 0.69), revealing that of the 70 pollen types found in the samples, 22 were shared by the two bee species in large proportions, 28 were exploited exclusively by Melipona seminigra merrillae and 20 were collected only by Melipona interrupta.     

Comparative resistance in Buckfast and Canadian stocks of honey bees (Apis mellifera L.) to infestation by honey bee tracheal mites (Acarapis woodi (Rennie))

The results of bioassay and colony evaluation demonstrated that British and Texas Buckfast honey bee stocks had one-third to one-half the mean prevalence and abundance of tracheal mites as Canadian standard stock, indicating that Buckfast stocks are less susceptible to tracheal mites than Canadian standard stock. Hybrid Canadian and Buckfast stocks exhibited resistance characteristics similar to only one of their parental stocks, suggesting the colony has an unknown effect on the expression of a bee's resistance to mites. A high correlation (r _s=0.66) between abundance values from the bioassay and colony evaluations indicates that the bioassay can be used to screen bees for mite resistance.     

Adaptive spatial biases in nectar deposition in the nests of honey bees

Honey bees store their large supply of honey at the top and, to a lesser extent, along the edges of their nests. Whether this pattern is the result of preferences for where nectar is deposited is unclear. Camazine, in a path breaking study of pattern formation, found evidence that workers deposit nectar at random, while Free and Williams in an earlier study found evidence that bees prefer to deposit nectar into particular types of comb. Here we reexamine this question. We tested three hypotheses, all of which posit that bees have adaptive biases which allow them to deposit nectar directly into the most useful locations. We found that bees have preferences for depositing nectar into cells at the top of the nest, into old vs. new comb, and into interior facing comb vs. comb facing the exterior environment. These preferences may ensure that nectar is placed directly into those regions where it will ultimately be stored. They may also result in an optimal pattern of comb usage with respect to brood rearing. This work, along with the results of previous studies, suggests that comb usage patterns may reflect competing selective pressures and be more complicated than previously thought. Received 17 May 2007; revised 15 June 2007; accepted 3 July 2007.