Time-activity budgets and space structuring by the different life stages ofVarroa jacobsoni in capped brood of the honey bee,Apis mellifera

Varroa jacobsoni reproduces in honey bee brood cells. Here the behavioral activity and use of space by infestingVarroa females and progeny were quantified in transparent artificial brood cells. The time-activity budget of both infesting and developing mites converged toward a stable pattern which was established during the bee prepupal stage of the infesting mites and the protonymphal stage of mite progeny. The pattern was such that infesting females and offspring eventually divided their activity between the fecal accumulation on the cell wall, which served as the rendezvous site for newly molted individuals, and the feeding site prepared on the pupa by the foundress. Other parts of the cell wall were used for oviposition and molting, away from the fecal accumulation on which activity of mobile stages was concentrated. Space structuring and the time-activity budget inVarroa probably evolved to enhance the number of fertilized females produced within the capped brood, where space and time are limiting factors. These behavioral adaptations parallel those of other mite species which show group behavior within cavities.     

Phenotypic and Genetic Analyses of the Varroa Sensitive Hygienic Trait in Russian Honey Bee (Hymenoptera: Apidae) Colonies

Varroa destructor continues to threaten colonies of European honey bees. General hygiene, and more specific Varroa Sensitive Hygiene (VSH), provide resistance towards the Varroa mite in a number of stocks. In this study, 32 Russian (RHB) and 14 Italian honey bee colonies were assessed for the VSH trait using two different assays. Firstly, colonies were assessed using the standard VSH behavioural assay of the change in infestation of a highly infested donor comb after a one-week exposure. Secondly, the same colonies were assessed using an “actual brood removal assay” that measured the removal of brood in a section created within the donor combs as a potential alternative measure of hygiene towards Varroa-infested brood. All colonies were then analysed for the recently discovered VSH quantitative trait locus (QTL) to determine whether the genetic mechanisms were similar across different stocks. Based on the two assays, RHB colonies were consistently more hygienic toward Varroa-infested brood than Italian honey bee colonies. The actual number of brood cells removed in the defined section was negatively correlated with the Varroa infestations of the colonies (r² = 0.25). Only two (percentages of brood removed and reproductive foundress Varroa) out of nine phenotypic parameters showed significant associations with genotype distributions. However, the allele associated with each parameter was the opposite of that determined by VSH mapping. In this study, RHB colonies showed high levels of hygienic behaviour towards Varroa -infested brood. The genetic mechanisms are similar to those of the VSH stock, though the opposite allele associates in RHB, indicating a stable recombination event before the selection of the VSH stock. The measurement of brood removal is a simple, reliable alternative method of measuring hygienic behaviour towards Varroa mites, at least in RHB stock.     

A comparison of the reproductive ability of <Emphasis Type="Italic">Varroa destructor</Emphasis> (Mesostigmata:Varroidae) in worker and drone brood of Africanized honey bees (<Emphasis Type="Italic">Apis mellifera</Emphasis>)

Colony infestation by the parasitic mite, Varroa destructor is one of the most serious problems for beekeeping worldwide. In order to reproduce varroa females, enter worker or drone brood shortly before the cell is sealed. To test the hypothesis that, due to the preference of mites to invade drone brood to reproduce, a high proportion of the mite reproduction should occur in drone cells, a comparative study of mite reproductive rate in worker and drone brood of Africanized honey bees (AHB) was done for 370 mites. After determining the number, developmental stage and sex of the offspring in worker cells, the foundress female mite was immediately transferred into an uninfested drone cell. Mite fertility in single infested worker and drone brood cells was 76.5 and 79.3%, respectively. There was no difference between the groups (X ² = 0.78, P = 0.37). However, one of the most significant differences in mite reproduction was the higher percentage of mites producing viable offspring (cells that contain one live adult male and at least one adult female mite) in drone cells (38.1%) compared to worker cells (13.8%) (X ² = 55.4, P < 0.01). Furthermore, a high level of immature offspring occurred in worker cells and not in drone cells (X ² = 69, P < 0.01). Although no differences were found in the percentage of non-reproducing mites, more than 74% (n = 85) of the mites that did not reproduce in worker brood, produced offspring when they were transferred to drone brood.     

Brood cell size of <Emphasis Type="Italic">Apis</Emphasis> <Emphasis Type="Italic">mellifera</Emphasis> modifies the reproductive behavior of <Emphasis Type="Italic">Varroa</Emphasis> <Emphasis Type="Italic">destructor</Emphasis>

We undertook a field study to determine whether comb cell size affects the reproductive behavior of Varroa destructor under natural conditions. We examined the effect of brood cell width on the reproductive behavior of V. destructor in honey bee colonies, under natural conditions. Drone and worker brood combs were sampled from 11 colonies of Apis mellifera. A Pearson correlation test and a Tukey test were used to determine whether mite reproduction rate varied with brood cell width. Generalized additive model analysis showed that infestation rate increased positively and linearly with the width of worker and drone cells. The reproduction rate for viable mother mites was 0.96 viable female descendants per original invading female. No significant correlation was observed between brood cell width and number of offspring of V. destructor. Infertile mother mites were more frequent in narrower brood cells.     

Selection for outbreeding in Varroa parasitising resistant honey bee (Apis mellifera) colonies

Parasitism is expected to select for counter‐adaptations in the host: driving a coevolutionary arms race. However, human interference between honey bees (Apis mellifera) and Varroa mites removes the effect of natural selection and restricts the evolution of host counter‐adaptations. With full‐sibling mating common among Varroa, this can rapidly select for virulent, highly inbred, Varroa populations. We investigated how the evolution of host resistance could affect the infesting population of Varroa mites. We screened a Varroa‐resistant honey bee population near Toulouse, France, for a Varroa resistance trait: the inhibition of Varroa's reproduction in drone pupae. We then genotyped Varroa which had co‐infested a cell using microsatellites. Across all resistant honey bee colonies, Varroa's reproductive success was significantly higher in co‐infested cells but the distribution of Varroa between singly and multiply infested cells was not different from random. While there was a trend for increased reproductive success when Varroa of differing haplotypes co‐infested a cell, this was not significant. This suggests local mate competition, through the presence of another Varroa foundress in a pupal cell, may be enough to help Varroa overcome host resistance traits; with a critical mass of infesting Varroa overwhelming host resistance. However, the fitness trade‐offs associated with preferentially co‐infesting cells may be too high for Varroa to evolve a mechanism to identify already‐infested cells. The increased reproductive success of Varroa when co‐infesting resistant pupal cells may act as a release valve on the selective pressure for the evolution of counter resistance traits: helping to maintain a stable host–parasite relationship.     

Fertility and reproductive rate of Varroa mite,Varroa destructor,in native and exotic honeybee,Apis mellifera L., colonies under Saudi Arabia conditions

Varroa mite is the most destructive pest to bee colonies worldwide. In Saudi Arabia, preliminary data indicated high infestation levels in the exotic honeybee colonies; such as Apis mellifera carnica and Apis mellifera ligustica, compared to native honeybee subspecies Apis mellifera jemenitica, which may imply higher tolerance to Varroasis. In this study, fertility and reproductive rate of Varroa mite, Varroa destructor, in capped brood cells of the native honeybee subspecies were investigated and compared with an exotic honeybee subspecies, A. m. carnica. Mite fertility was almost alike (87.5% and 89.4%) in the native and craniolan colonies respectively. Similarly, results did not show significant differences in reproduction rate between both subspecies (F = 0.66, Pr > F = 0.42). Number of adult Varroa daughters per fertile mother mite was 2.0 and 2.1 for native and craniolan honeybee subspecies respectively. This may indicate that mechanisms of keeping low infestation rates in the native honeybee colonies are not associated with Varroa reproduction. Therefore, potential factors of keeping lower Varroa infestation rates in native honey bee subspecies should be further investigated.     

Differential periods ofVarroa mite invasion into worker and drone cells of honey bees

Invasion ofVarroa mites into honeybee brood cells was studied in an observation hive, using combs with cell openings at one side only. The cell bottoms had been replaced by a transparent sheet, through which mites were clearly visible after invasion into a cell. Mites invaded worker cells from 15–20 h preceding cell capping, whereas they invaded drone cells from 40–50 h preceding capping. The larger number of mites generally found in drone cells, when compared to worker cells, may be partly due to the longer period of mite invasion into drone brood.     

Varroa Mite Infestations in Elevated Honey Bee Brood Cells: Effects of Context and Caste

The Varroa mite infestation level of honey bee, Apis mellifera, worker larvae reared in individual raised cells was 6-fold higher than in the adjacent six cells surrounding them; this differential infestation rate is similar to published values of higher mite infestations of drone cells compared to worker cells. Infestation levels in control cells were the same as in the surrounding cells. In contrast to infestation of these individually raised cells, Varroa mites invaded worker larvae in raised cells along the perimeter of a patch of raised cells (10 by 21 rows) 2.5 times more often than surrounding unraised cells, and similarly ca. 2.5 times more often than in the remaining raised cells (interior) of this patch. In similarly prepared frames of drone comb, Varroa mites invaded individually raised drone cells 3.3-fold more often than the adjacent surrounding cells and control cells. On the other hand, Varroa mites infested drone larvae in the interior of the raised-patch area as often as drones in raised cells along the perimeter of the raised-patch, and this rate was ca. 2.5-fold higher than for drone larvae in unraised cells surrounding the raised-patch and drone larvae in control cells. The higher levels of infestation of raised cells did not come at the expense of the surrounding cells, i.e., the infestation levels of the adjacent surrounding cells were the same as in control cells. For worker larvae, the increased number of mites invading individual raised cells and edge cells of the raised patch were proportional to the number of surrounding nonraised cells. The relationship between raised cell-edges, observations of mite walking behavior on comb surfaces, and larval-to-cell-rim distances are discussed in relation to their possible roles in eliciting mite invasion of honey bee larval cells and contrasted to the putative role of kairomones in larval-host location.     

Non-reproducing Varroa jacobsoni Oud. in honey bee worker cells—status of mites or effect of brood cells?

The parasitic mite Varroa jacobsoni Oud. reproduces in sealed honey bee brood cells. Within worker cells a considerable fraction of the mites do not produce offspring. It is investigated whether variation in the ratio of cells without reproduction is caused by properties of the worker brood, or by the state of the mites entering cells. Pieces of brood comb were taken from colonies of 12 different bee lines and were placed simultaneously into highly infested colonies. Non-reproduction was independent of the origin of the brood pieces, indicating a minor role of a variation due to different brood origin. Between colonies used for infestation, however, it differed considerably. A comparison of the proportion of cells without reproduction when infested by one Varroa mite or when infested by two or three Varroa mites showed, that non-reproduction was mainly related to the state of the mites entering cells, and only to a minor degree to an influence of the brood cells. A high ratio of worker cells without reproduction was consistently reported in bee lines which survive the disease without treatment, and a high level of non-reproduction is thus regarded to be a key factor in breeding bees for high Varroa tolerance. The current results indicate, that differences in this trait are only to a minor degree related to differences between bee lines in the ability of the bee brood to induce oviposition. These differences seem rather to depend on other, unknown colony factors influencing the reproductive state of Varroa when they enter cells for reproduction.     

Reproduction of <Emphasis Type="Italic">Varroa destructor</Emphasis> and offspring mortality in worker and drone brood cells of Africanized honey bees

Varroa destructor is known to be the most serious parasite of Apis mellifera worldwide. In order to reproduce varroa females enter worker or drone brood shortly before the cell is sealed. From March to December 2008, the reproductive rate and offspring mortality (mature and immature stages), focusing on male absence and male mortality of V. destructor, was investigated in naturally infested worker and drone brood of Africanized honey bees (AHB) in Costa Rica. Data were obtained from 388 to 403 single infested worker and drone brood cells, respectively. Mite fertility in worker and drone brood cells was 88.9 and 93.1%, respectively. There was no difference between the groups (X² = 3.6, P = 0.06). However, one of the most significant differences in mite reproduction was the higher percentage of mites producing viable offspring in drone cells (64.8%) compared to worker cells (37.6%) (X² = 57.2, P < 0.05). A greater proportion of mites in worker brood cells produced non-viable female offspring. Mite offspring mortality in both worker and drone cells was high in the protonymph stage (mobile and immobile). A significant finding was the high rate of male mortality. The worker and drone brood revealed that 23.9 and 6.9%, respectively, of the adult male offspring was found dead. If the absence (missing) of the male and adult male mortality are taken together the percentage of cells increased to 40.0 and 21.3% in worker and drone cells, respectively (X² = 28.8, P < 0.05). The absence of the male or male mortality in a considerable number of worker cells naturally infested with varroa is the major factor in our study which reduces the production of viable daughters in AHB colonies in Costa Rica.     

Differential viral levels and immune gene expression in three stocks of Apis mellifera induced by different numbers of Varroa destructor

The viral levels and immune responses of Italian honey bees (IHB), Russian honey bees (RHB) and an outcross of Varroa Sensitive Hygienic bees (POL) deliberately infested with one or two foundress Varroa were compared. We found that the Deformed wing virus (DWV) level in IHB inoculated with one or two foundress Varroa increased to about 10³ or 10⁵ fold the levels of their uninfested brood. In contrast, POL (10² or 10⁴ fold) and RHB (10² or l0⁴ fold) supported a lower increase in DWV levels. The feeding of different stages of Varroa nymphs did not increase DWV levels of their pupal hosts. Analyses of their corresponding Varroa mites showed the same trends: two foundress Varroa yielded higher DWV levels than one foundress, and the addition of nymphs did not increase viral levels. Using the same pupae examined for the presence of viruses, 16 out of 24 genes evaluated showed significant differential mRNA expression levels among the three honey bee stocks. However, only four genes (Defensin, Dscam, PPOact and spaetzle), which were expressed at similar levels in uninfested pupae, were altered by the number of feeding foundress Varroa and levels of DWV regardless of stocks. This research provides the first evidence that immune response profiles of different honey bee stocks are induced by Varroa parasitism.     

Development of early infestations by the miteVarroa jacobsoni in honey-bee (Apis mellifera) colonies in cold climates

The development of an infestation by five to eight introduced adult females ofVarroa jacobsoni Oud. in 35 honey-bee (Apis mellifera L.) colonies was monitored for 16 months with no outside source of infestation. Calculations on the size of the mite populations were based on collection of debris, samples of bees and brood, and estimates of number of bees and broodcells during the summer. In the winter, only dead bees and debris were collected. Samples were taken at 3-week intervals. Data indicated that the mite population probably could increase more than 100 times within one summer, and more than ten times between years, in a climate with a brood-rearing period of less than five months. A large variation in mite population increase existed between colonies. The winter mortality of mites that die with the host or drop from the winter cluster has a large influence on the population dynamics of the mite. Data also indicated that the simple method of counting mites in hive debris is a useful parameter for monitoring the population development ofVarroa in colonies with hatching brood.     

Ontogenesis of the mite Varroa jacobsoni Oud. in drone brood of the honeybee Apis mellifera L. under natural conditions

A study carried out during the summer of 1994, in southern England, investigated the developmental times and mortality ofVarroa jacobsoni inApis mellifera drone cells. The position and time of capping of 2671 naturally infested drone cells were recorded. Six hours after the cell was capped, 90% of the mites were free from the brood food to start feeding on the developing drone. The developmental time of the mite's first three female offspring (133±3 h) and the male offspring (150 h) and the intervals between egg laying (20–32 h) were similar to those found in worker cells. However, the mortality of the offspring was much lower in drone cells than worker cells. The mode numbers of eggs laid were six and five in drone and worker cells, respectively. All offspring had ample time to develop fully in drone cells with the sixth offspring reaching maturity approximately 1 day before the drone bee emerged. Normal mites (those which lay five or six viable eggs) produced on average four female adult offspring but since only around approximately 55% of the mite population produced viable offspring the mean number of viable adult female offspring per total number of mother mites was 2 to 2.2 in drone cells.     

Reproduction of Varroa destructor in worker brood of Africanized honey bees (Apis mellifera)

Reproduction and population growth of Varroa destructor was studied in ten naturally infested, Africanized honeybee (AHB) (Apis mellifera) colonies in Yucatan, Mexico. Between February 1997 and January 1998 monthly records of the amount of pollen, honey, sealed worker and drone brood were recorded. In addition, mite infestation levels of adult bees and worker brood and the fecundity of the mites reproducing in worker cells were determined. The mean number of sealed worker brood cells (10,070 ± 1,790) remained fairly constant over the experimental period in each colony. However, the presence and amount of sealed drone brood was very variable. One colony had drone brood for 10 months and another for only 1 month. Both the mean infestation level of worker brood (18.1 ± 8.4%) and adult bees (3.5 ± 1.3%) remained fairly constant over the study period and did not increase rapidly as is normally observed in European honey bees. In fact, the estimated mean number of mites fell from 3,500 in February 1997 to 2,380 in January 1998. In May 2000 the mean mite population in the study colonies was still only 1,821 mites. The fertility level of mites in this study was much higher (83–96%) than in AHB in Brazil(25–57%), and similar to that found in EHB (76–94%). Mite fertility remained high throughout the entire study and was not influenced by the amount of pollen, honey or worker brood in the colonies. This revised version was published online in July 2006 with corrections to the Cover Date.     

Comparison of diagnostic methods for detection of low infestation levels ofVarroa jacobsoni in honey-bee (Apis mellifera) colonies

Thirty-five honey-bee colonies, originally free fromVarroa jacobsoni (Oudemans) were monitored approximately every third week for the presence of the mite during 16 months following an initial introduction of five to eight adultVarroa females in early July. Investigations of hive debris detected the presence ofV. jacobsoni in 22 colonies (63%) within three months of the mite introduction. During the first winter period (October–April), mites were found in the hive debris of 13 colonies (37%). In terms of detectingVarroa during the summer in colonies with sealed brood, investigations of hive debris were more effective than sampling of brood. Brood sampling was more effective than sampling of live bees. In colonies without sealed brood, investigations of hive debris or of live bee samples seemed approximately equally efficient. The highest correlation between sampling methods was found between daily mite downfall and mites per live bee (r=0.81) in colonies with sealed brood. During the winter, investigations of dead bees and hive debris were approximately equally efficient in detectingVarroa.     

Does time spent on adult bees affect reproductive success of Varroa mites?

Reproduction ofVarroa jacobsoni Oudemans (Acari: Varroidae) and the number ofVarroa mites that were found dead on the bottom board of the hive, were studied in relation to the period the mites spent on adult honey bees,Apis mellifera L. (Hymenoptera: Apidae), prior to invasion into brood cells. The maximum period on adult bees was 23 days. To introduce mites, combs with emerging worker brood, heavily infested with mites, were placed into a colony and removed the next day. At the beginning of the first day following emergence from brood cells, 18% of the mites introduced into the colony was found on the bottom of the hive. Part of these mites may already have died inside the capped brood cells, and then fallen down after cleaning of cells by the bees. At the second and third day following emergence, respectively 4% and 2% of the mites on adult bees at the previous day was recovered on the bottom, whereas from the fourth day on only 0.6% of the mites on adult bees was recovered on the bottom per day. After invasion into brood cells, 8–12% of the mites did not produce any offspring. Of the mites that did reproduce, the total number of offspring was 4.0–4.4 per mite during one reproductive cycle, part of which may reach maturity resulting in 1.2–1.3 viable daughters, and 8–10% of the mites produced only male offspring. Reproduction was independent of the period the mites had spent on adult bees prior to invasion into brood cells.     

Impact of the Phoretic Phase on Reproduction and Damage Caused by Varroa destructor (Anderson and Trueman) to Its Host,the European Honey Bee (Apis mellifera L.)

Varroa destructor is a parasitic mite of the honeybee that causes thousands of colony losses worldwide. The parasite cycle is composed of a phoretic and a reproductive phase. During the former, mites stay on adult bees, mostly on nurses, to feed on hemolymph. During the latter, the parasites enter brood cells and reproduce. We investigated if the type of bees on which Varroa stays during the phoretic phase and if the duration of this stay influenced the reproductive success of the parasite and the damage caused to bees. For that purpose, we used an in vitro rearing method developed in our laboratory to assess egg laying rate and the presence and number of fully molted daughters. The expression level of two Varroa vitellogenin genes (VdVg1 and VdVg2), known to vary throughout reproduction, was also quantified. Results showed that the status of the bees or time spent during the phoretic phase impacts neither reproduction parameters nor the Varroa vitellogenin genes levels of expression. However, we correlated these parameters to the gene expression and demonstrated that daughters expressed the vitellogenin genes at lower levels than their mother. Regarding the damage to bees, the data indicated that a longer stay on adult bees during the phoretic phase resulted in more frequent physical deformity in newborn bees. We showed that those mites carry more viral loads of the Deformed Wing Virus and hence trigger more frequently overt infections. This study provides new perspectives towards a better understanding of the Varroa-honeybee interactions.     

Factors influencing host choice of the honey bee parasite Varroa jacobsoni Oud

Reproducing Varroa jacobsoni obtained from brood cells of Apis mellifera L. with 13–16 day old bees (pupae) and Varroa mites kept on adult bees for at least 8 days were simultaneously tested for their choice in three host types. Comparisons were made of attractiveness of Varroa jacobsoni to nurse bees, pollen foragers as to larvae from nearly capped brood cells. Host choices were observed in Petri dishes and in an Y-shaped olfactometer. Varroa jacobsoni obtained from capped brood cells showed a stronger preference for nurse bees in Petri dish simultaneous choice tests with pollen foragers or larvae than did mites which were previously kept on adult bees. In olfactometer simultaneous choice tests, the two mite test groups showed no clear difference in preferences for bees of different ages. The preference of Varroa jacobsoni for bees of different ages is therefore not only influenced by host factors but also by intrinsic factors in female mites that depend on the mite's reproductive stage.     

Number of reproductive cycles of Varroa jacobsoni in honey-bee (Apis mellifera) colonies

Very little data exists concerning the number of reproductive cycles performed by individual Varroa mites. To understand the population dynamics of the Varroa mite it is necessary to know the number of fertile female offspring each Varroa female produces during her lifetime. The lifetime reproduction capacity of the mite consists of the mean number of fertile female offspring produced during each reproductive cycle multiplied by the mean number of cell passages. This paper describes an experimental design to estimate the number of reproductive cycles where mites are transferred to new mite-free colonies for reproduction in sealed brood cells. The data presented suggests that the mean number of reproductive cycles performed by the individual female mite is larger than previously accepted. Under optimal conditions, the mean number of reproductive cycles by Varroa females is probably greater than 1.5 but less than 2. Furthermore, the results show that the reproductive success of Varroa females going into cells to reproduce is not influenced by previous brood cycles.     

Insemination Capacity and Dispersal in Relation to Sex Allocation Decisions in Goniozus legneri (Hymenoptera: Bethylidae): Why Are There More Males in Larger Broods?

Models considering sex ratio optima under single foundress strict local mate competition predict that female bias will be reduced by stochasticity in sex allocation, developmental mortality of males and limited insemination capacity of males. In all three cases the number of males per brood is expected to increase with brood size. Sex ratio optima may also be less female biased when several mothers contribute offspring to local mating groups or if non‐local mating occurs between members of different broods; again more males are expected in larger broods. In the parasitoid wasp Goniozus legneri (Hymenoptera: Bethylidae), sex allocation has only a small stochastic component, developmental mortality is low and non‐siblings are unlikely to develop in the same brood. However, the number of males per brood increases with the size of the brood (produced by a single mother). We investigated the further possibilities of limited insemination capacity and non‐local mating using a naturalistic experimental protocol. We found that limited insemination capacity is an unlikely general explanation for the increase in number of males with brood size. All males and females dispersed from both mixed and single sex broods. Although most females in mixed sex broods mated prior to dispersal, these data suggest that non‐local mating is possible, for instance via male immigration to broods containing virgin females. This may influence sex ratio optima and account for the trend in male number.