Numerous studies have recently reported on the discovery of bee viruses in different arthropod species and their possible transmission routes, vastly increasing our understanding of these viruses and their distribution. Here, we review the current literature on the recent advances in understanding the transmission of viruses, both on the presence of bee viruses in Apis and non-Apis bee species and on the discovery of previously unknown bee viruses. The natural transmission of bee viruses will be discussed among different bee species and other insects. Finally, the research potential of in vivo (host organisms) and in vitro (cell lines) serial passages of bee viruses is discussed, from the perspective of the host-virus landscape changes and potential transmission routes for emerging bee virus infections.

Western honey bees, Apis mellifera, live in highly eusocial colonies that are each typically headed by a single queen. The queen is the sole reproductive female in a healthy colony, and because long-term colony survival depends on her ability to produce a large number of offspring, queen health is essential for colony success. Honey bees have recently been experiencing considerable declines in colony health. Among a number of biotic and abiotic factors known to impact colony health, disease and queen failure are repeatedly reported as important factors underlying colony losses. Surprisingly, there are relatively few studies on the relationship and interaction between honey bee diseases and queen quality. It is critical to understand the negative impacts of pests and pathogens on queen health, how queen problems might enable disease, and how both factors influence colony health. Here, we review the current literature on queen reproductive potential and the impacts of honey bee parasites and pathogens on queens. We conclude by highlighting gaps in our knowledge on the combination of disease and queen failure to provide a perspective and prioritize further research to mitigate disease, improve queen quality, and ensure colony health.

SummaryDiseases are known to be one of the major contributors to colony losses. Within a Europe-wide experiment on genotype -environment interactions, an initial 621 colonies were set up and maintained from 2009 to 2012. The colonies were monitored to investigate the occurrence and levels of key pathogens. These included the mite Varroa destructor (mites per 10 g bees), Nosema spp. (spore loads and species determination), and viruses (presence/absence of acute bee paralysis virus (ABPV) and deformed wing virus (DWV)). Data from 2010 to the spring of 2011 are analysed in relation to the parameters: genotype, environment, and origin (local vs. non-local) of the colonies in the experiment. The relative importance of different pathogens as indicators of colony death within the experiment is compared. In addition, pathogen occurrence rates across the geographic locations are described. 216Meixner et al.

Deformed wing virus is an important contributor to honey bee colony losses. Frequently queen failure is reported as a cause for colony loss. Here we examine whether sexual transmission during multiple matings of queens is a possible way of virus infection in queens. In an environment with high prevalence of deformed wing virus, queens (n = 30) were trapped upon their return from natural mating flights. The last drone’s endophallus (n = 29), if present, was removed from the mated queens for deformed wing virus quantification, leading to the detection of high-level infection in 3 endophalli. After oviposition, viral quantification revealed that seven of the 30 queens had high-level deformed wing virus infections, in all tissues, including the semen stored in the spermathecae. Two groups of either unmated queens (n = 8) with induced egg laying, or queens (n = 12) mated in isolation with drones showing comparatively low deformed wing virus infections served as control. None of the control queens exhibited high-level viral infections. Our results demonstrate that deformed wing virus infected drones are competitive to mate and able to transmit the virus along with semen, which occasionally leads to queen infections. Virus transmission to queens during mating may be common and can contribute noticeably to queen failure.

Deformed wing virus (DWV) is an important pathogen in a broad range of insects, including honey bees. Concordant with the spread of Varroa, DWV is present in the majority of honey bee colonies and can result in either low-level infections with asymptomatic bees that nonetheless exhibit increased colony loss under stress, or high-level infections with acute effects on bee health and viability. DWV can be transmitted vertically or horizontally and evidence suggests that horizontal transmission via Varroa is associated with acute symptomatic infections. Vertical transmission also occurs and is presumably important for the maintenance of DWV in honey bee populations. To further our understanding the vertical transmission of DWV through queens, we performed three experiments: we studied the quantitative effectiveness of vertical transmission, surveyed the prevalence of successful egg infection under commercial conditions, and distinguished among three possible mechanisms of transmission. We find that queen-infection level predicts the DWV titers in their eggs, although the transmission is not very efficient. Our quantitative assessment of DWV demonstrates that eggs in 1/3 of the colonies are infected with DWV and highly infected eggs are rare in newly-installed spring colonies. Additionally, our results indicate that DWV transmission occurs predominantly by virus adhering to the surface of eggs (transovum) rather than intracellularly. Our combined results suggest that the queens’ DWV vectoring capacity in practice is not as high as its theoretical potential. Thus, DWV transmission by honey bee queens is part of the DWV epidemic with relevant practical implications, which should be further studied.

Social evolution has led to distinct life‐history patterns in social insects, but many colony‐level and individual traits, such as egg size, are not sufficiently understood. Thus, a series of experiments was performed to study the effects of genotypes, colony size and colony nutrition on variation in egg size produced by honey bee (Apis mellifera) queens. Queens from different genetic stocks produced significantly different egg sizes under similar environmental conditions, indicating standing genetic variation for egg size that allows for adaptive evolutionary change. Further investigations revealed that eggs produced by queens in large colonies were consistently smaller than eggs produced in small colonies, and queens dynamically adjusted egg size in relation to colony size. Similarly, queens increased egg size in response to food deprivation. These results could not be solely explained by different numbers of eggs produced in the different circumstances but instead seem to reflect an active adjustment of resource allocation by the queen in response to colony conditions. As a result, larger eggs experienced higher subsequent survival than smaller eggs, suggesting that honey bee queens might increase egg size under unfavourable conditions to enhance brood survival and to minimize costly brood care of eggs that fail to successfully develop, and thus conserve energy at the colony level. The extensive plasticity and genetic variation of egg size in honey bees has important implications for understanding life‐history evolution in a social context and implies this neglected life‐history stage in honey bees may have trans‐generational effects.

SummaryHygienic behaviour in the honey bee, Apis mellifera, is the uncapping and removal of dead, diseased or infected brood from sealed cells by worker bees. We determined the effect of hygienic behaviour on varroa population growth and incidence of deformed wing virus (DWV), which can be transmitted by varroa. We treated 42 broodless honey bee colonies with oxalic acid in early January 2013 to reduce varroa populations to low levels, which we quantified by extracting mites from a sample of worker bees. We quantified varroa levels, again when the colonies were broodless, 48 weeks later. During the summer the hygienic behaviour in each colony was quantified four times using the Freeze Killed Brood (FKB) removal assay, and ranged from 27.5 % to 100 %. Varroa population increased greatly over the season, and there was a significant negative correlation between varroa increase and FKB removal. This was entirely due to fully hygienic colonies with >95 % FKB having only 43 % of the varroa build up of the less hygienic colonies. None of the 14 colonies with >80 % FKB removal had overt symptoms of DWV, whilst 36 % of the less hygienic colonies did. Higher levels of FKB removal also correlated significantly with lower numbers of DWV RNA copies in worker bees, but not in varroa mites. On average, fully hygienic colonies had c. 10,000 times less viral RNA than less hygienic colonies. Hacia el control integrado de varroa: efecto de la variación en el comportamiento higiénico entre colonias de abejas de la miel con aumento de la población de ácaros y de la incidencia de virus de las alas deformadas ResumenEl comportamiento higiénico en la abeja de la miel, Apis mellifera, se basa en el desoperculado y la eliminación de la cría muerta, enferma o infectada a de las celdas selladas por las abejas obreras. Se determinó el efecto del comportamiento higiénico en el crecimiento de la población de varroa y la incidencia del virus de las alas deformadas (VAD), que puede ser transmitido por la varroa. Se han tratado 42 colonias de abejas de la miel sin larvas con ácido oxálico a principios de enero de 2013 para reducir las poblaciones de varroa a niveles bajos, lo que fue cuantificado mediante la extracción de los ácaros de una muestra de las abejas obreras. Se cuantificaron los niveles de varroa, de nuevo cuando las colonias no tenían cría, 48 semanas después. Durante el verano, el comportamiento higiénico en cada colonia se cuantificó cuatro veces utilizando el ensayo de congelar la cría para matarla (BCM), y este varió entre el 27,5% y el 100%. La población de Varroa aumentó considerablemente durante la temporada, y se observó una correlación negativa significativa entre el aumento de la varroasis y la 556 Toufailia et al.

Honey bee virus prevalence data are an essential prerequisite for managing epidemic events in a population. A survey study was carried out for seven viruses in colonies representing a healthy Danish honey bee population. In addition, colonies from apiaries with high level Varroa infestation or high level of winter mortality were also surveyed. Results from RT-qPCR showed a considerable difference of virus levels between healthy and sick colonies. In the group of healthy colonies, no virus was detected in 36% of cases, while at least one virus was found in each of the sick colonies. Virus titers varied among the samples, and multiple virus infections were common in both groups with a high prevalence of Sacbrood virus (SBV), Black queen cell virus (BQCV) and Deformed wing virus (DWV). Based on the distribution of virus titers, we established four categories of infection: samples free of virus (C = 0), samples with low virus titer (estimated number of virus copies 0 < C < 103), samples with medium virus titer (103 ≤ C < 107) and samples with high virus titer (C ≥ 107). This allowed us to statistically compare virus levels in healthy and sick colonies. Using categories to communicate virus diagnosis results to beekeepers may help them to reach an informed decision on management strategies to prevent further spread of viruses among colonies.