Current Results

WP1 - WP2 - WP3 - WP4 - WP5 - WP6 - WP7 - WP8

Current Results: Workpackage 1

Forsgren E, Fries I (2010):
Comparative virulence of Nosema ceranae and Nosema apis in individual European honey bees. VETERINARY PARASITOLOGY 170: 212-217

Nosema apis and Nosema ceranae are intracellular microsporidian parasites infecting the midgut epithelial cells of adult honey bees. N. ceranae was considered to be restricted to the Asian honey bee, Apis cerana, but is nowadays a parasite found also in the European honey bee (Apis mellifera) across most of the world. Recent surveys and experimental work suggest that N. ceranae is a serious threat to the global beekeeping industry. It has been suggested that N. ceranae induces significantly higher mortality in honey bees than N. apis, but little is known about their comparative virulence. In this study, we used in vivo infection experiments to study the two parasites' different virulence (i.e. multiplication rate and infectivity). A qPCR was developed to elucidate within host competition between the two parasites using mixed infections. The outcome of the experiments indicates minor differences in infectious dose and multiplication rate between the two species. Moreover, the mortality caused by N. ceranae was not significantly higher than for N. apis and N. ceranae appeared to have no competitive advantage within host.

Current Results: Workpackage 2

Gauthier L, Ravallec M, Tournaire M, Cousserans F, Bergoin M, Dainat B, de Miranda JR (2011):
Viruses Associated with Ovarian Degeneration in Apis mellifera L. Queens. PLOS ONE 6: e16217

Queen fecundity is a critical issue for the health of honeybee (Apis mellifera L.) colonies, as she is the only reproductive female in the colony and responsible for the constant renewal of the worker bee population. Any factor affecting the queen's fecundity will stagnate colony development, increasing its susceptibility to opportunistic pathogens. We discovered a pathology affecting the ovaries, characterized by a yellow discoloration concentrated in the apex of the ovaries resulting from degenerative lesions in the follicles. In extreme cases, marked by intense discoloration, the majority of the ovarioles were affected and these cases were universally associated with egg-laying deficiencies in the queens. Microscopic examination of the degenerated follicles showed extensive paracrystal lattices of 30 nm icosahedral viral particles. A cDNA library from degenerated ovaries contained a high frequency of deformed wing virus (DWV) and Varroa destructor virus 1 (VDV-1) sequences, two common and closely related honeybee Iflaviruses. These could also be identified by in situ hybridization in various parts of the ovary. A large-scale survey for 10 distinct honeybee viruses showed that DWV and VDV-1 were by far the most prevalent honeybee viruses in queen populations, with distinctly higher prevalence in mated queens (100% and 67%, respectively for DWV and VDV-1) than in virgin queens (37% and 0%, respectively). Since very high viral titres could be recorded in the ovaries and abdomens of both functional and deficient queens, no significant correlation could be made between viral titre and ovarian degeneration or egg-laying deficiency among the wider population of queens. Although our data suggest that DWV and VDV-1 have a role in extreme cases of ovarian degeneration, infection of the ovaries by these viruses does not necessarily result in ovarian degeneration, even at high titres, and additional factors are likely to be involved in this pathology.

Current Results: Workpackage 3

Alaux C, Folschweiller M, McDonnell C, Beslay D, Cousin M, Dussaubat C, Brunet JL, Le Conte Y (2011):
Pathological effects of the microsporidium Nosema ceranae on honey bee queen physiology (Apis mellifera). JOURNAL OF INVERTEBRATE PATHOLOGY 106: 380-385

Nosema ceranae, a microsporidian parasite originally described in the Asian honey bee Apis cerana, has recently been found to be cross-infective and to also parasitize the European honey bee Apis mellifera. Since this discovery, many studies have attempted to characterize the impact of this parasite in A. mellifera honey bees. Nosema species can infect all colony members, workers, drones and queens, but the pathological effects of this microsporidium has been mainly investigated in workers, despite the prime importance of the queen, who monopolizes the reproduction and regulates the cohesion of the society via pheromones. We therefore analyzed the impact of N. ceranae on queen physiology. We found that infection by N. ceranae did not affect the fat body content (an indicator of energy stores) but did alter the vitellogenin titer (an indicator of fertility and longevity), the total antioxidant capacity and the queen mandibular pheromones, which surprisingly were all significantly increased in Nosema-infected queens. Thus, such physiological changes may impact queen health, leading to changes in pheromone production, that could explain Nosema-induced supersedure (queen replacement).

Current Results: Workpackage 4

Behrens D, Huang Q, Gessner C, Rosenkranz P, Frey E, Locke B, Moritz RFA, Kraus FB (2011):
Three QTL in the honey bee Apis mellifera L. suppress reproduction of the parasitic mite Varroa destructor. ECOLOGY AND EVOLUTION 1: 451-458

Varroa destructor is a highly virulent ectoparasitic mite of the honey bee Apis mellifera and a major cause of colony losses for global apiculture. Typically, chemical treatment is essential to control the parasite population in the honey bee colony. Nevertheless a few honey bee populations survive mite infestation without any treatment. We used one such Varroa mite tolerant honey bee lineage from the island of Gotland, Sweden, to identify quantitative trait loci (QTL) controlling reduced mite reproduction. We crossed a queen from this tolerant population with drones from susceptible colonies to rear hybrid queens. Two hybrid queens were used to produce a mapping population of haploid drones. We discriminated drone pupae with and without mite reproduction, and screened the genome for potential QTL using a total of 216 heterozygous microsatellite markers in a bulk segregant analysis. Subsequently, we fine mapped three candidate target regions on chromosomes 4, 7, and 9. Although the individual effect of these three QTL was found to be relatively small, the set of all three had significant impact on suppression of V. destructor reproduction by epistasis. Although it is in principle possible to use these loci for marker-assisted selection, the strong epistatic effects between the three loci complicate selective breeding programs with the Gotland Varroa tolerant honey bee stock.

Current Results: Workpackage 5

De Smet L, Ravoet J, de Miranda JR, Wenseleers T, Mueller MY, Moritz RFA, de Graaf DC (2012):
BeeDoctor, a Versatile MLPA-Based Diagnostic Tool for Screening Bee Viruses. PLOS ONE

The long-term decline of managed honeybee hives in the world has drawn significant attention to the scientific community and bee-keeping industry. A high pathogen load is believed to play a crucial role in this phenomenon, with the bee viruses being key players. Most of the currently characterized honeybee viruses (around twenty) are positive stranded RNA viruses. Techniques based on RNA signatures are widely used to determine the viral load in honeybee colonies. High throughput screening for viral loads necessitates the development of a multiplex polymerase chain reaction approach in which different viruses can be targeted simultaneously. A new multiparameter assay, called "BeeDoctor", was developed based on multiplex-ligation probe dependent amplification (MLPA) technology. This assay detects 10 honeybee viruses in one reaction. "BeeDoctor" is also able to screen selectively for either the positive strand of the targeted RNA bee viruses or the negative strand, which is indicative for active viral replication. Due to its sensitivity and specificity, the MLPA assay is a useful tool for rapid diagnosis, pathogen characterization, and epidemiology of viruses in honeybee populations. "BeeDoctor" was used for screening 363 samples from apiaries located throughout Flanders; the northern half of Belgium. Using the "BeeDoctor", virus infections were detected in almost eighty percent of the colonies, with deformed wing virus by far the most frequently detected virus and multiple virus infections were found in 26 percent of the colonies.

Current Results: Workpackage 6

Locke B, Fries I (2011):
Characteristics of honey bee colonies (Apis mellifera) in Sweden surviving Varroa destructor infestation. APIDOLOGIE 42: 533-542

A population of European honey bees (Apis mellifera) surviving Varroa destructor mite infestation in Sweden for over 10 years without treatment, demonstrate that a balanced host-parasite relationship may evolve over time. Colony-level adaptive traits linked to Varroa tolerance were investigated in this population to identify possible characteristics that may be responsible for colony survival in spite of mite infestations. Brood removal rate, adult grooming rate, and the mite distribution between brood and adults were not significantly different in the untreated population compared with treated control colonies. However, colony size and the reproductive success of the mite were significantly reduced in surviving colonies compared with control colonies. Our data suggest that colony-level adaptive traits may limit mite population growth by reducing mite reproduction opportunities and also by suppressing the mite reproductive success.

Current Results: Workpackage 7

Truchado P, Ponce A, Tomás-Barberán FA, Allende A (2011):
Selected Phytochemical Bioactive Compounds as Quorum Sensing Inhibitors. Acta Horticulturae (ISHS) 939:93-97

Bacterial infections remain an important problem for human health. The control of bacterial infections has been traditionally treated by inhibiting microbial growth using different types of antibiotics. However, the ability of different bacteria to resist the inhibitory action of antibiotics has become a global problem. In fact, there is an important need for the development of new antimicrobials that act on novel bacterial targets. Many pathogenic bacteria control their population and regulate gene expression in response to their cell population density using diffusible signalling com¬pounds. This type of communication has been referred to as "quorum sensing" (QS). This phenomenon can be essential for the synchronization of the virulence production factors, which make it an attractive therapeutic target. Therefore, the search of non-toxic compounds, which inhibit QS and so, the virulence of pathogenic bacteria can bring new alternatives for the treatment of bacterial infections in humans.
In this work, we made an attempt to screen the anti-QS activity of 11 bioactive compounds extracted from fruits and vegetables using the biosensor strain, Chromobacterium violaceum. The anti-QS activity was determined quantifying the violacein production of the biosensor strain at three concentrations (50, 100 and 200 µg/ml). At least five of the tested compounds (cinnamaldehyde, pomegranate extracts, ellagic acid resveratrol and rutin) showed anti-QS activity against the biosensor strain. The obtained results showed the potential of bioactive compounds extracted from fruits and vegetables to be used as a new category of anti-pathogenic compounds against bacterial infections.

Current Results: Workpackage 8

Aebi A, Neumann P (2011):
Endosymbionts and honey bee colony losses? TRENDS IN ECOLOGY & EVOLUTION 26: 494