Localization of Deformed Wing Virus (DWV) in the Brains of Apis mellifera (European Honey Bees)

The purpose of the current research project is to design a successful in-situ hybridization to identify regions within the brains of honeybees where DWV replicates. The localization of the virus in the brains of the bees can draw a connection between CCDand DWV.In conclusion, these results demonstrate that in bees infected with DWV the virus replicates actively in very important regions of the brain, including neuropils that are responsible for vision and olfaction. This means that the virus could adversely affect the vision and olfaction of the honeybees making it difficult for bees to behave normally.

Nectar distribution and its relation to food quality in honeybee (Apis mellifera) colonies

In honeybees (Apis niellifera), the process of nectar collection is considered a straightforward example of task partitioning with two subtasks or two intersecting cycles of activity: (1) foraging and (2) storing of nectar, linked via its transfer between foragers and food processors. Many observations suggest, however, that nectar colleclion and processing in honeybees is a complex process, involving workers of other sub-castes and depending on variables such as resource profitability or the amount of stored honey. It has been observed that food processor bees often distribute food to other hive bees after receiving it from incoming foragers, instead of storing it immediately in honey cells. While there is little information about the sub-caste affiliation and the behaviour of these second-order receivers, this stage may be important for the rapid distribution of nutrients and related information. To investigate the identity of these second-order receivers, we quantified behaviours following nectar transfer and compared these behaviours with the behaviour of average worker hive-bees. Furthermore, we tested whether food quality (sugar concentration) affects the behaviour of the second-order receivers. Of all identified second-order receivers, 59.3% performed nurse duties, 18.5% performed food-processor duties and 22.2% performed forager duties. After food intake, these bees were more active, had more trophallaxes (especially offering contacts) compared to average workers and they were found mainly in the brood area, independent of food quality. Our results show that the liquid food can be distributed rapidly among many bees of the three main worker sub-castes, without being stored in honey cells first. Furthermore, the results suggest that the rapid distribution of food partly depends on the high activity of second-order receivers.

Sugar concentration influences decision making in <i>Apis mellifera</i> L. workers during early-stage honey storage behaviour

Decision making in honeybees is based on in- formation which is acquired and processed in order to make choices between two or more al- ternatives. These choices lead to the expression of optimal behaviour strategies such as floral constancy. Optimal foraging strategies such as floral constancy improve a colony’s chances of survival, however to our knowledge, there has been no research on decision making based on optimal storage strategies. Here we show, using diagnostic radioentomology, that decision mak- ing in storer bees is influenced by nectar sugar concentrations and that, within 48 hours of col- lection, honeybees workers store carbohydrates in groups of cells with similar sugar concentra- tions in a nonrandom way. This behaviour, as evidenced by patchy spatial cell distributions, would help to hasten the ripening process by reducing the distance between cells of similar sugar concentrations. Thus, colonies which ex- hibit optimal storage strategies such as these would have an evolutionary advantage and im- prove colony survival expectations over less efficient colonies and it should be plausible to select colonies that exhibit these preferred traits.

Genome characterization, prevalence and distribution of a Macula-like virus from Apis mellifera and Varroa destructor

Around 14 distinct virus species-complexes have been detected in honeybees, each with one or more strains or sub-species. Here we present the initial characterization of an entirely new virus species-complex discovered in honeybee (Apis mellifera L.) and varroa mite (Varroa destructor) samples from Europe and the USA. The virus has a naturally poly-adenylated RNA genome of about 6500 nucleotides with a genome organization and sequence similar to the Tymoviridae (Tymovirales; Tymoviridae), a predominantly plant-infecting virus family. Literature and laboratory analyses indicated that the virus had not previously been described. The virus is very common in French apiaries, mirroring the results from an extensive Belgian survey, but could not be detected in equally-extensive Swedish and Norwegian bee disease surveys. The virus appears to be closely linked to varroa, with the highest prevalence found in varroa samples and a clear seasonal distribution peaking in autumn, coinciding with the natural varroa population development. Sub-genomic RNA analyses show that bees are definite hosts, while varroa is a possible host and likely vector. The tentative name of Bee Macula-like virus (BeeMLV) is therefore proposed. A second, distantly related Tymoviridae-like virus was also discovered in varroa transcriptomes, tentatively named Varroa Tymo-like virus (VTLV).

Polinização por Apis mellifera em soja em soja transgênica [Glycine max (L.) Merrill] Merrill] Roundup ReadyTM cv. BRS 245 RR e convencional cv. BRS 133.

RESUMO: O objetivo deste estudo foi verificar a influência de Apis mellifera na produção de grãos e qualidade de sementes da soja transgênica Glycine max (L.) Merrill Roundup ReadyTM e convencional. A soja transgênica foi plantada intercalada com a convencional, em 18 parcelas, em três tratamentos: gaiolas com abelhas A. mellifera, gaiolas sem abelhas e áreas descobertas, com livre visitação de insetos. Na soja transgênica, em três parcelas de cada tratamento, foi aplicado glifosato, 30 dias após a emergência. Os parâmetros analisados foram: produção de grãos; número de vagens por planta; peso de 100 sementes e porcentagem de germinação das sementes. Não houve diferença entre as cultivares, entretanto a produção de 2.757,40 kg ha-1 obtida na área coberta por gaiola com abelhas, e 2.828,47 kg ha-1 na área livre para visitação de insetos foram superiores a 2.000,53 kg ha-1 da área coberta por gaiola sem abelhas. O número de vagens por planta foi maior na área coberta por gaiola com abelhas (38,28) e área livre (32,65), quando comparado com o da área coberta por gaiola sem abelhas (21,19). O peso médio de 100 sementes e a germinação das sementes não foram diferentes entre as cultivares e nem entre os tratamentos. Conclui-se que, para as cultivares estudadas, houve benefício na produção de grãos de 37,84%, quando foi permitida a visita de abelhas. ABSTRACT. Pollination by Apis mellifera in transgenic soy (Glycine max (L.) Merrill) Roundup Ready? cv. BRS 245 RR and conventional cv. BRS 133. This research was carried out to evaluate the influence of Africanized honeybees in grain production and seed quality of Glycine max (L.) Merrill Roundup Ready? transgenic soy, as well as of conventional soy. Transgenic soy was interposed with conventional soybean, in 18 plots and three treatments: covered area with Africanized honeybees, covered area without honeybees, and uncovered area with free insect visitation. The herbicide Glyphosate was applied on three plots of each treatment of transgenic soy, 30 days after emergence. Grain production, number of pods/plant, weight per 100 seeds, and seed germination percentage were evaluated. There was no difference among cultivars; however, the production in the covered area with honeybees (2757.4 kg ha-1) and in the uncovered area (2828.47 kg ha-1) were higher than in the covered area without honeybees (2000.53 kg ha-1). The number of pods/plant was greater than in the covered area with honeybees (38.28) and in the uncovered area (32.65) as compared to the covered area without honeybees (21.19). The weight per 100 seeds seed germination did not differ among cultivars or treatments. It can be concluded that, for these cultivars, there was a rise in grain production of 37.84% when honeybee visits were allowed.

巨尾阿丽蝇(Aldrichina grahami）和意大利蜂(Apis mellifera L.)在自由状态下电击回避学习能力的研究

本能和学习能力是动物生存所需的两种基本行为方式，它们之间存在一定的拮抗关系。巨尾阿丽蝇是一种具有强大生存本能的双翅目昆虫，而该物种是否具有良好的学习能力是本研究所关心的问题。而蜜蜂作为神经生物学研究中的模式生物，已有各种行为学范式对其进行了研究，包括束缚状态下的伸喙反射（PER）、螫刺反射（SER），及自由状态下与糖水结合的偏好性学习模式，但是对于自由状态下的回避反应尚没有研究。因此，本研究借鉴了啮齿类被动回避学习研究的范式，设计并制作了一个适用于研究昆虫在自由状态下的电击回避学习能力的装置，利用诱饵与电刺激相结合的方式对巨尾阿丽蝇和蜜蜂在自由状态下的电击回避学习能力进行了研究。研究得到以下结果：一、巨尾阿丽蝇能够学会回避此负性刺激，且当电压范围在5到45 V之间时有显著的回避行为，而电压达到60 V时则受到明显伤害。二、蜜蜂在被电击之后的采蜜频率有显著的降低。同样说明，它们能够学会对电刺激进行回避。有趣的是，是否有蜜蜂被电击后在电板上挣扎的情况也是影响它们采蜜频率的因素之一。三、本论文中所采用的实验范式可以成为一种新的对自由状态下昆虫电击回避学习能力进行研究的实验范式。

Grooming and hygienic behaviours of the honey bee (Apis mellifera) toward the parasitic mite Varroa destructor

Despite a multitude of environmental stressors, the Varroa mite is still regarded as the greatest cause of honey bee mortality in its invaded range. Breeding honey bees that are resistant to the mite is an important area of research. This thesis aimed to gain a better understanding of the grooming and hygienic behaviours of Russian honey bees (RHB). The effect of a break in the synchrony of a mite’s life cycle on reproductive success was tested through brood inoculation experiments. Mites released by hygienic behaviour and forced to enter a new cell are less likely to lay male offspring. Through laboratory cage assays it was found that daughter mites are more susceptible to grooming behaviour. A new method of marking Varroa mites was developed which would enable a single cohort of mites to be followed after inoculation. A strong brood removal trait was noticed in RHB colonies, therefore they were tested for Varroa sensitive hygienic (VSH) behaviour. RHB demonstrated levels of VSH as high as the USDA line bred specifically for this behaviour. In addition the same QTL found to be responsible for the trait in VSH bees, was associated with VSH in RHB stock. Previous work showed that the ratio of older mites to total trapped mites (O/T) in the debris of honey bee colonies demonstrated the strongest association with colony infestation. This research showed that O/T is associated with VSH and brood removal behaviour. In addition, bees that displayed high levels of VSH in this study were also more likely to spend a longer amount of time grooming in laboratory assays. This indicates that both grooming and hygienic behaviours play important roles in the resistance of RHB stock. Their likelihood to be expressed by other stocks is discussed and recommendations for further research are provided.

Nosema ceranae has infected Apis mellifera in Europe since at least 1998 and may be more virulent than Nosema apis

Nosema ceranae, a microsporidian formerly regarded as confined to its Asiatic host Apis cerana, has recently been shown to parasitise Apis mellifera and to have spread throughout most of the world in the past few years. Using a temporal sequence of N = 28 Nosema isolates from Finland from 1986-2006, we now find (i) that N. ceranae has been present in Europe since at least 1998 and (ii) that it has increased in frequency across this time period relative to Nosema apis, possibly leading to higher mean spore loads per bee. We then present results of a single laboratory infection experiment in which we directly compare the virulence of N. apis with N. ceranae. Though lacking replication, our results suggest (iii) that both parasites build up to equal numbers per bee by day 14 post infection but that (iv) N. ceranae induces significantly higher mortality relative to N. apis.

Ovariole number - a predictor of differential reproductive success among worker subfamilies in queenless honeybee (Apis mellifera L.) colonies

A honeybee queen normally mates with 10-20 drones, and reproductive conflicts may arise among a colony's different worker patrilines, especially after a colony has lost its single queen and the workers commence egg laying. In this study, we employed microsatellite markers to study aspects of worker reproductive competition in two queenless Africanized honeybee colonies. First, we determined whether there was a bias among worker patrilines in their maternity of drones and, second, we asked whether this bias could be attributed to differences in the degree of ovary activation of workers. Third, we relate these behavioral and physiological factors to ontogenetic differences between workers with respect to ovariole number. Workers from each of three (colony A) and one (colony B) patrilineal genotypes represented less than 6% of the worker population, yet each produced at least 13% of the drones in a colony, and collectively they produced 73% of the drones. Workers representing these genotypes also had more developed follicles and a greater number of ovarioles per ovary. Across all workers, ovariole development and number were closely correlated. This suggests a strong effect of worker genotype on the development of the ovary already in the postembryonic stages and sets a precedent to adult fertility, so that

Does infection by Nosema ceranae cause "Colony Collapse Disorder" in honey bees (Apis mellifera)?

Nosema ceranae is an emergent and potentially virulent pathogen of the honey bee (Apis mellifera) that has spread across the world in the last 10 or so years. Its precise origin and timing of spread are currently unclear because of a lack of appropriate genetic markers and inadequate sampling in putative Asian source populations. Though it has been dismissed as a cause of CCD in the USA based on correlational analyses of snapshot sampling of diseased hives, observations of naturally infected colonies suggest that it leads to colony collapse in Spain. Experiments are sorely needed to investigate its impact on individuals and colonies, and to pin down a causal relationship between N. ceranae and colony collapse. Whether N. ceranae is displacing N. apis is uncertain. For temperate zone apiculturalists, global climate change may mean that N. ceranae presents more of a challenge than has hitherto been considered the case.

Deformed wing virus and drone mating flights in the honey bee (Apis mellifera): implications for sexual transmission of a major honey bee virus

Deformed wing virus (DWV) represents an ideal model to study the interaction between mode of transmission and virulence in honey bees since it exhibits both horizontal and vertical transmissions. However, it is not yet clear if venereal-vertical transmission represents a regular mode of transmission for this virus in natural honey bee populations. Here, we provide clear evidence for the occurrence of high DWV titres in the endophallus of sexually mature drones collected from drone congregation areas (DCAs). Furthermore, the endophallus DWV titres of drones collected at their maternal hives were no different from drones collected at nearby DCAs, suggesting that high-titre DWV infection of the endophallus does not hinder the ability of drones to reach the mating area. The results are discussed within the context of the dispersal of DWV between colonies and the definition of DWV virulence with respect to the transmission route and the types of tissues infected.

Honey bee (Apis mellifera) venom induces AIM2 inflammasome activation in human keratinocytes

Following allergen exposure, cytokines and other pro-inflammatory signals play an important role in the immunological cascade leading to allergic sensitization. Inflammasomes sense exogenous and endogenous danger signals and trigger IL-1β and IL-18 activation which in turn shape Th2 responses. Honey bee venom (BV) allergies are very common; however, the local inflammatory cascade leading to the initiation of allergic sensitization is poorly understood. In this study, the local inflammatory cascades in skin after exposure to BV were investigated.