Behaviour of Varroa mites invading honey bee brood cells

Invasion behaviour of Varroa jacobsoni into honey bee brood cells was studied using an observation hive. The mites were carried close to a suitable brood cell by the bees. Subsequently, the mites moved from the bees to the rim of the cell, walked quickly inside, crawled between the larva and the cell wall, and moved onto the bottom of the cell. Varroa mites were never seen walking across the comb, and entering and leaving brood cells as has been described for Tropilaelaps clareae. Differences in invasion strategies between V. jacobsoni and T. clareae are discussed.     

Effect of host brood type on the number of offspring laid by the honeybee parasite Varroa jacobsoni

The number of offspring laid by individual mites, varies depending on the type (drone or worker) of honeybee brood cell invaded. The number of offspring laid by individual mites increases when artificially transferred from worker to drone brood and vice versa when moved in the opposite direction.     

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.     

Further observations on the correlation between attractiveness of honey bee brood cells toVarroa jacobsoni and the distance from larva to cell rim

Varroa jacobsoni Oudemans (Acari: Varroidae) was studied with respect to invasion into different types of honeybee,Apis mellifera L., brood cells. Different cell types were obtained by shortening and elongating of cells, grafting worker larvae into drone cells andvice versa. The type of cell strongly affected the number of mites per cell, and the attractive period of the cells to the mites. The type of cell also affected the distance from larva to cell rim preceding cell capping. When this distance was larger in comparison to control cells of the same age, the attractive period of the brood cells was shorter andvice versa. Since in all cell types the distance from larva to cell rim continuously decreased preceding cell capping, this negative correlation is in agreement with the hypothesis that there is a critical larva-rim distance under which brood cells are attractive to mites. Then, the length of the attractive period of brood cells depends on the moment this critical distance is reached. The distribution of mites over different cell types in turn results from differences in the attractive period.     

The mite Varroa jacobsoni does not transmit American foulbrood from infected to healthy colonies

The present study was conducted to determine whether Varroa jacobsoni can transmit American foulbrood (AFB), caused by the bacterium Paenibacillus larvae to healthy colonies by the surface transport of spores. Five two-storey Langstroth colonies of Apis mellifera ligustica were infested by placing a sealed brood comb, with 10% Varroa prevalence, between the central brood combs of each colony. Two months later the colonies were inoculated with P. larvae by adding brood comb pieces with clinical signs of AFB (45±5 scales per colony). After 60 days the brood area was completely uncapped by means of dissecting needles and tweezers, separating the Varroa mites from the larvae and the collected mites were introduced at a rate of 51 per colony into four recipient hives placed in an isolated apiary. Twenty female Varroa specimens were separated at random and observed by SEM. Paenibacillus larvae spores were found on the dorsal shield surface and on idiosomal setae. All colonies died after 4–5 months due to a high incidence of varroosis. No clinical AFB symptoms or P. larvae spores were observed in microscopic preparations. It is concluded that Varroa jacobsoni does not transmit AFB from infected to healthy colonies; it does, however transport P. larvae spores on its surface.     

The role of trophallaxis in the distribution of Perizin in a honeybee colony with regard to the control of the Varroa mite

It is claimed that Perizin, a pesticide to control the mite Varroa jacobsoni, acts systemically and is distributed by trophallaxis of the bees. We studied the role of trophallactic interactions in the distribution of coumaphos, the active ingredient, among the colony members and whether coumaphos can reach all mites by systemic activity. Colonies were divided into three compartments by a screen, one receiving a Perizin treatment by sprinkling, the others receiving no Perizin. In this way it was possible to trace the amount of coumaphos transferred between bees through the screen from the treated part to the untreated one by trophallaxis. After different periods of time the number of fallen mites was counted and the amount of coumaphos in bees was determined for all hive compartments. We found that trophallactic interactions are of minor importance in the distribution of Perizin between the two compartments. The recommended method of sprinkling Perizin over the bees was shown to be very inefficient; only 24% of the applied amount reaches the alimentary canal of the bees; the rest must therefore remain at other places: on the outside of the bees, in the combs and on the hive-parts.     

Varroa jacobsoni (Acari: Varroidae) is more than one species

Varroa jacobsoni was first described as a natural ectoparasitic mite of the Eastern honeybee (Apis cerana) throughout Asia. It later switched host to the Western honeybee (A. mellifera) and has now become a serious pest of that bee worldwide. The studies reported here on genotypic, phenotypic and reproductive variation among V. jacobsoni infesting A. cerana throughout Asia demonstrate that V. jacobsoni is a complex of at least two different species. In a new classification V. jacobsoni is here redefined as encompassing nine haplotypes (mites with distinct mtDNA CO-I gene sequences) that infest A. cerana in the Malaysia–Indonesia region. Included is a Java haplotype, specimens of which were used to first describe V. jacobsoni at the beginning of this century. A new name, V. destructor n. sp., is given to six haplotypes that infest A. cerana on mainland Asia. Adult females of V. destructor are significantly larger and less spherical in shape than females of V. jacobsoni and they are also reproductively isolated from females of V. jacobsoni. The taxonomic positions of a further three unique haplotypes that infest A. cerana in the Philippines is uncertain and requires further study.Other studies reported here also show that only two of the 18 different haplotypes concealed within the complex of mites infesting A. cerana have become pests of A. mellifera worldwide. Both belong to V. destructor, and they are not V. jacobsoni. The most common is a Korea haplotype, so-called because it was also found parasitizing A. cerana in South Korea. It was identified on A. mellifera in Europe, the Middle East, Africa, Asia, and the Americas. Less common is a Japan/Thailand haplotype, so-called because it was also found parasitizing A. cerana in Japan and Thailand. It was identified on A. mellifera in Japan, Thailand and the Americas.Our results imply that the findings of past research on V. jacobsoni are applicable mostly to V. destructor. Our results will also influence quarantine protocols for bee mites, and may present new strategies for mite control.     

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.     

First data on resistance mechanisms of Varroa jacobsoni (OUD.) against tau-fluvalinate

In 1991, the first losses of efficacy of tau-fluvalinate against the honeybee ectoparasite Varroa jacobsoni Oud. were recorded in Sicily. Since then, diminished efficacy with available pyrethroid treatments has been encountered in many regions of Italy. The aim of this study was to investigate the type of resistance in V. jacobsoni to the pyrethroid tau-fluvalinate by focusing on metabolic resistance mechanisms (detoxication). After developing a suitable application method, two synergists were used: piperonyl butoxide (PBO), as an inhibitor of the microsomal monooxygenases of the cytochrome P450 complex and S,S,S-tributylphosphorotrithioate (DEF), which blocks esterases. A significant decrease in the LC₅₀ values of the susceptible and of the resistant mite strains after the application of PBO was observed. A slight decrease of the LC₅₀ values was also observed after the application of DEF. However, this decrease was not significant. These results indicate that the resistance of Varroa mites to tau-fluvalinate can partly be explained by an increased detoxication due to the monooxygenases in the P450 system, which is blocked by PBO. Esterases seems to play a negligible role. Whether glutathione-S-transferases are involved, is still unknown, but other mechanisms, such as the modification of the binding sites and/or reduced uptake might be involved as well.     

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.     

The presence of inhibitors of the reproduction of Varroa jacobsoni Oud. (Gamasida: Varroidae) in infested cells

The mite Varroa jacobsoni was reared in artificial gelatin cells under laboratory conditions and the possible presence of factors inhibiting Varroa reproduction was studied. In cells infested with three mites, the mean offspring per female was reduced to 75% of that in singly infested cells. When gelatin cells were used for two successive rearing cycles, both the proportion of reproducing females and the offspring per reproducing female were significantly lower in cells that had contained an infested larva during the first rearing cycle than in those with an uninfested larva. The mean reduction of the offspring per female was 48%; this suggests that inhibitors of the reproduction are released into infested cells. Treatment of gelatin cells with the hexane extract of cells in which an infested bee pupa had developed caused a 21% reduction in the mean offspring per female, with a difference close to the significance level (p=0.07).     

Number of adult female mites Varroa jacobsoni Oud. on hive debris from honey bee colonies artificially infested to monitor mite population increase (Mesostigmata: Varroidae)

Six honey bee colonies hived in Langstroth nuclei were each artificially infested with 100 phoretic Varroa mites. Hive debris on bottom inserts was inspected every 3–4 days. The adult Varroa mites were compared with mounted specimens and catalogued into lightly pigmented and darkly pigmented females. After 4 months, an acaricide treatment was used to estimate the final mite population. Based on light and dark adult counts, we propose a balancing equation that follows the Varroa population increase at 7 day intervals and allows the calculation of experimental population growth rates. The calculated Varroa finite rate of increase is =1.021. Exponential and logistic growth models fitted to the balancing equation data gave R ²=0.986 and R ²=0.991, respectively. To develop a more precise model it would be necessary to follow the population growth beyond our experimental data.     

Synergistic and molecular interactions between the formamidine amitraz and copper(II) sulfate,used against the mite Varroa jacobsoni O., a parasite of the honeybee Apis mellifera L.

Toxicological field assays have shown that the shock-treatment efficacy of the formamidine pesticide amitraz, used against the parasitic mite Varroa jacobsoni, is synergistically improved by the administration of copper(II) sulfate through feeding of the honeybees. Amitraz is autoxidized and this process is accompanied by chemiluminescence. The emission is enhanced in the presence of low concentrations of H₂O₂. A dose-related inhibition of the chemiluminescence by CuSO₄ was observed; consistent with the formation of copper-amitraz complexes evidenced in vitro. The results suggest the possibility that a protection of amitraz by cupric ions might be at the origin of the enhancement of its toxicity and thus makes a contribution to the observed synergy.     

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.     

Methyl palmitate: a novel product of the Medfly (Ceratitis capitata) corpus allatum

The corpus allatum (CA) of adult female Ceratitis capitata produces methyl palmitate (MP) in vitro, in addition to JHB₃ and JH III. Biosynthesized MP migrates on TLC and co-elutes from RP-18 HPLC with synthetic MP. Its identity is verified herein by GCMS. MP production is up-regulated twofold by mevastatin, an inhibitor of mevalonic acid-dependent isoprene biosynthesis. Fosmidomycin, an inhibitor of mevalonic acid-independent isoprene synthesis in graminaceous plants, up-regulates MP synthesis by about fourfold. However, it does not depress JHB₃ biosynthesis concurrently. This suggests that the initial enzyme(s) in the conversion of 1-deoxy-xylulose 5-phosphate to isoprene is presumably present in C. capitata, but is inhibited by fosmidomycin, and this inhibition diverts precursors to MP synthesis. Phytol, an acyclic diterpene, might be suppressing isoprene biosynthesis by CA, thereby resulting in a fourfold increase in the MP biosynthesis. Linolenic acid is an end-product and its presence in incubation media up-regulates MP biosynthesis by twofold, presumably due to the feedback diversion to biosynthesis of C_16:0 and its methyl ester. Biosynthesis of MP is markedly depressed after mating, while otherwise maintained at significantly higher levels in virgin females. MP biosynthesis is significantly reduced in virgin females by direct axonal control but is less consistent after mating.     

Distribution of deformed wing virus within honey bee (Apis mellifera) brood cells infested with the ectoparasitic mite Varroa destructor

The distribution of deformed wing virus (DWV) in adult female Varroa destructor and in their progeny in relation to the pupal host bee was investigated to evaluate acquisition and transfer of DWV by the mites. The results clearly show that adult female mites regularly act as competent vectors of DWV, however, they do not acquire or transfer virus on all possible occasions. Mother mites may contain DWV while the pupal host remains free from overt infection and both mother mites and mite progeny may not acquire detectable amounts of DWV from an infected host bee. However, a majority of mites feeding on pupae that emerge with deformed wings will contain DWV. The data also demonstrates that both adult and immature mite progeny most likely acquire DWV from DWV-infected host bees and not from their mother mites. Possible explanations for the obtained results are discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

Survival of the mite Varroa jacobsoni Oud. (Mesostigmata: Varroidae) in broodless colonies of the honey bee Apis mellifera L. (Hymenoptera: Apidae)

Varroa mite free colonies of the honey bee Apis mellifera L. were artificially infested, with either parasitized bees or infested worker brood. Queens were kept in cages to provide broodless conditions during the experiment. Parasites that fell to the bottom of the hive were monitored at 3–4 days intervals for three months. An acaricide treatment was used to recover mites still alive after this time period. Survivorship at each interval was calculated and life table functions of the phoretic mite cohorts were obtained. Trends in survival of Varroa cohorts showed maximum lifespans ranging from 80 to 100 days. Life expectancy of these phoretic cohorts at the beginning of the experiment ranges between 19 to 41, with a mean of 31 days.     

Positive anemotaxis by Varroa mites: responses to bee odour plumes and single clean‐air puffs

Abstract.The stimuli and mechanisms mediating host location and host choice by the bee mite, Varroa jacobsoni (Oudemans), are currently unknown. It is shown that Varroa can use single clean‐air puffs and bee‐odour plumes in a wind tunnel as directional cues. Varroa turned nearly straight upwind in response to single 0.1‐s puffs of clean air directed at 90° to the their anterior‐posterior axis. They turned significantly further to their left side (104°) than to their right (76°), but showed no difference in latency to initiation of the turns (means of 63.3 ms vs. 62.6 ms, respectively). They also followed bee‐odour plumes in a wind tunnel. When released in odour and control plumes mid‐way between the plume's origin and the downwind end of the tunnel, mites responding to bee‐odour walked upwind in, or along the edge of, the odour plume with 38% making contact with the odour delivery tube; mites in clean air did not walk upwind along the air stream, and none made contact with the air delivery tube. Walking speeds were not different between the bee‐odour and control groups (0.28 vs. 0.29 cm s^–1); there were also no differences in the turning rates (96.85 vs. 97.16 deg s^–1 and 388.08 vs. 379.18 deg cm^–1, respectively). Under all conditions, mites walked in a zigzag fashion.