The reproductive program of female Varroa destructor mites is triggered by its host, Apis mellifera

Reproducing Varroa females begin oviposition on a host larva by laying an unfertilized (male) egg, followed by fertilized (female) offspring. Using transfer experiments, we examined whether the sequence of sexes in the brood cell is triggered by a host stimulus. When reproducing Varroa females were transferred from white-eyed pupae (worker brood) into freshly capped worker brood cells, 77% (n = 22 fertile mites after the transfer) began a new reproductive cycle by laying a male egg. The proportion of brood cells with male offspring was similar to naturally infested brood cells. Varroa females transferred into brood cells with young pupae reproduced, but only 6% (n = 16 fertile mites after the transfer) produced male offspring. This was significantly different from male production in naturally reproducing Varroa females and those transferred into freshly capped brood cells. We conclude that a host stimulus present in freshly capped brood cells triggers both the start of reproduction and the sequence of sexes.     

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.     

蜜蜂巢房大小影响狄斯瓦螨的繁殖行为

在具有相同类型幼虫的雄蜂和工蜂巢房中,人工接入狄斯瓦螨Varroa destructorAnderson&Trueman,比较巢房大小不同,对于螨繁殖的影响。结果显示:狄斯瓦螨在具有工蜂幼虫的工蜂房(WW)中的繁殖率为94.4%,而在具有工蜂幼虫的雄蜂房(WD)中繁殖率只有27.7%,差异极显著。在具有工蜂幼虫的工蜂房中,每只雌螨产出后代的平均数为3.35±1.56只;在具有工蜂幼虫的雄蜂房中每只雌螨产出后代的平均数为0.49±0.93只,差异极显著。表明:在具有相同类型幼虫存在的情况下,狄斯瓦螨喜欢较小的巢房,狄斯瓦螨在较小巢房中的繁殖能力明显高于较大的巢房。     

Behavior of varroa mites in worker brood cells of Africanized honey bees

The ectoparasitic mite Varroa destructor is currently the most important pest of the honey bee, Apis mellifera. Because mite reproduction occurs within the sealed cell, the direct observation of varroa activity inside the cell is difficult. A video observation method using transparent polystyrol cells containing infested brood was used to analyze the behavior of varroa mites in worker brood of Africanized honey bees. We recorded how mites feed on the larva and pupa, construct a fecal accumulation site and how the bee larva carried out some longitudinal movements around the cell. The feeding activity of the foundress mite varies during the course of the cycle. On the prepupa mites were found to feed often (0.3 ± 0.2 bouts h⁻¹) for a period of 8.7 ± 8.4 min h⁻¹ and there was no preference for a specific segment as feeding site. On the opposite, during the pupal stage mites fed less often (0.1 ± 0.1 bouts h⁻¹) for a period of 6.2 ± 4.0 min h⁻¹ and almost always at a particular site (92.4%). On pupa, 83.7% of the feeding was on the 2nd abdominal segment (n = 92), and only few perforations were found on the thorax. Varroa shows a preference for defecation in the posterior part of the cell (cell apex), close to the bee′s anal zone. We found a high correlation between the position of the feeding site on the pupa and the position of the fecal accumulation on the cell wall. Most infested cells have only one fecal accumulation site and it was the favorite resting site for the mite, where it spent 24.3 ± 3.9 min h⁻¹. Longitudinal displacements were observed in 28.0% (n = 25) of the analyzed bee larvae. Turning movements around the cell, from the bottom to the top, were carried out by these larvae, mainly during the second day (47.7 ± 22.5 min h⁻¹), just before pupation, with a total time of 874.9 ± 262.2 min day⁻¹ (n = 7 individuals). These results in worker brood of Africanized bees demonstrate adaptations of varroa mites to parasitizing the developing bee inside the capped brood cells.     

Comparative reproduction of <Emphasis Type="Italic">Varroa destructor</Emphasis> in different types of Russian and Italian honey bee combs

Earlier studies showed that Russian honey bees support slow growth of varroa mite population. We studied whether or not comb type influenced varroa reproduction in both Russian and Italian honey bees, and whether Russian bees produced comb which inhibited varroa reproduction. The major differences found in this study concerned honey bee type. Overall, the Russian honey bees had lower (2.44 ± 0.18%) levels of varroa infestation than Italian honey bees (7.20 ± 0.60%). This decreased infestation resulted in part from a reduced number of viable female offspring per foundress in the Russian (0.85 ± 0.04 female) compared to the Italian (1.23 ± 0.04 females) honey bee colonies. In addition, there was an effect by the comb built by the Russian honey bee colonies that reduced varroa reproduction. When comparing combs having Russian or Italian colony origins, Russian honey bee colonies had more non-reproducing foundress mites and fewer viable female offspring in Russian honey bee comb. This difference did not occur in Italian colonies. The age of comb in this study had mixed effects. Older comb produced similar responses for six of the seven varroa infestation parameters measured. In colonies of Italian honey bees, the older comb (2001 dark) had fewer (1.13 ± 0.07 females) viable female offspring per foundress than were found in the 2002 new (1.21 ± 0.06 females) and 1980s new (1.36 ± 0.08 females) combs. This difference did not occur with Russian honey bee colonies where the number of viable female offspring was low in all three types of combs. This study suggests that honey bee type largely influences growth of varroa mite population in a colony.     

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.     

Semiochemicals Influencing the Host-finding Behaviour of Varroa Destructor

Studies of Varroa destructor orientation to honey bees were undertaken to isolate discrete chemical compounds that elicit host-finding activity. Petri dish bioassays were used to study cues that evoked invasion behaviour into simulated brood cells and a Y-tube olfactometer was used to evaluate varroa orientation to olfactory volatiles. In Petri dish bioassays, mites were highly attracted to live L5 worker larvae and to live and freshly freeze-killed nurse bees. Olfactometer bioassays indicated olfactory orientation to the same type of hosts, however mites were not attracted to the odour produced by live pollen foragers. The odour of forager hexane extracts also interfered with the ability of mites to localize and infest a restrained nurse bee host. Varroa mites oriented to the odour produced by newly emerged bees (<16 h old) when choosing against a clean airstream, however in choices between the odours of newly emerged workers and nurses, mites readily oriented to nurses when newly emerged workers were <3 h old. The odour produced by newly emerged workers 18–20 h of age was equally as attractive to mites as that of nurse bees, suggesting a changing profile of volatiles is produced as newly emerged workers age. Through fractionation and isolation of active components of nurse bee-derived solvent washes, two honey bee Nasonov pheromone components, geraniol and nerolic acid, were shown to confuse mite orientation. We suggest that V. destructor may detect relative concentrations of these compounds in order to discriminate between adult bee hosts, and preferentially parasitize nurse bees over older workers in honey bee colonies. The volatile profile of newly emerged worker bees also may serve as an initial stimulus for mites to disperse before being guided by allomonal cues produced by older workers to locate nurses. Fatty acid esters, previously identified as putative kairomones for varroa, proved to be inactive in both types of bioassays.     

Using an in vitro system for maintaining <Emphasis Type="Italic">Varroa destructor</Emphasis> mites on <Emphasis Type="Italic">Apis mellifera</Emphasis> pupae as hosts: studies of mite longevity and feeding behavior

Varroa destructor mites (varroa) are ectoparasites of Apis mellifera honey bees, and the damage they inflict on hosts is likely a causative factor of recent poor honey bee colony performance. Research has produced an arsenal of control agents against varroa mites, which have become resistant to many chemical means of their control, and other means have uncertain efficacy. Novel means of control will result from a thorough understanding of varroa physiology and behavior. However, robust knowledge of varroa biology is lacking; mites have very low survivability and reproduction away from their natural environment and host, and few tested protocols of maintaining mites in vitro are available as standardized methods for varroa research. Here, we describe the ‘varroa maintenance system’ (VMS), a tool for maintaining in vitro populations of varroa on its natural host, and present best practices for its use in varroa and host research. Additionally, we present results using the VMS from research of varroa and host longevity and varroa feeding behavior. Under these conditions, from two trials, mites lived an average of 12 and 14 days, respectively. For studies of feeding behavior, female mites inflicted wounds located on a wide range of sites on the host’s integument, but preferred to feed from the host’s abdomen and thorax. Originally in the phoretic-phase, female mites in VMS had limited reproduction, but positive instances give insights into the cues necessary for initiating reproduction. The VMS is a useful tool for laboratory studies requiring long-term survival of mites, or host–parasite interactions.     

Reproductive biology of <Emphasis Type="Italic">Varroa destructor</Emphasis> in Africanized honey bees (<Emphasis Type="Italic">Apis mellifera</Emphasis>)

Since its first contact with Apis mellifera, the population dynamics of the parasitic mite Varroa destructor varies from one region to another. In many regions of the world, apiculture has come to depend on the use of acaricides, because of the extensive damage caused by varroa to bee colonies. At present, the mite is considered to contribute to the recent decline of honey bee colonies in North America and Europe. Because in tropical climates worker brood rearing and varroa reproduction occurs all year round, it could be expected that here the impact of the parasite will be even more devastating. Yet, this has not been the case in tropical areas of South America. In Brazil, varroa was introduced more than 30 years ago and got established at low levels of infestation, without causing apparent damage to apiculture with Africanized honey bees (AHB). The tolerance of AHB to varroa is apparently attributable, at least in part, to resistance in the bees. The low fertility of this parasite in Africanized worker brood and the grooming and hygienic behavior of the bees are referred as important factors in keeping mite infestation low in the colonies. It has also been suggested that the type of mite influences the level of tolerance in a honey bee population. The Korea haplotype is predominant in unbalanced host-parasite systems, as exist in Europe, whereas in stable systems, as in Brazil, the Japan haplotype used to predominate. However, the patterns of varroa genetic variation have changed in Brazil. All recently sampled mites were of the Korea haplotype, regardless whether the mites had reproduced or not. The fertile mites on AHB in Brazil significantly increased from 56% in the 1980s to 86% in recent years. Nevertheless, despite the increased fertility, no increase in mite infestation rates in the colonies has been detected so far. A comprehensive literature review of varroa reproduction data, focusing on fertility and production of viable female mites, was conducted to provide insight into the Africanized bee host-parasite relationship.     

Changes in the reproductive ability of the mite Varroa destructor (Anderson e Trueman) in africanized honey bees (Apis mellifera L.) (Hymenoptera: Apidae) colonies in southern Brazil

Varroa destructor has been in Brazil for more than 30 years, but no mortality of honeybee colonies due to this mite has been recorded. Africanized bee infestation rates attained by varroa have been low, without causing measurable damage to Brazilian apiculture. The low reproductive ability of this parasite in Africanized bee worker brood cells has been considered an important factor for maintaining the host-parasite equilibrium. Nevertheless, the possible substitution of the haplotype of the mite Varroa destructor that has occurred recently in Brazil could affected the reproductive ability of the population of this parasite in Brazil. The reproductive ability of worker of the mite females was evaluated in over one thousand 17-18 day-old Africanized worker brood cells each of the two periods. The percentage of fertile mites increased from 56% in the 1980s to 86% in 2005-2006. The difference in the percentage of females that produced deutonymphs, female progeny that can reach the adult stage at bee emergence, was even greater. In 2005-2006, 72% of the females that invaded worker brood had left at the least one viable descendant, compared to 35% in 1986-1987.     

An experimental analysis of pleometrotic advantage in the desert seed-harvester antMessor pergandei (Hymenoptera; Formicidae)

Summary Starting colonies of the desert seed-harvester antMessor pergandei are clumped in the field and face severe intraspecific competition through brood raiding. Single foundress laboratory colonies ofM. pergandei are more likely to succeed at brood raiding with conspecific colonies if they are given additional workers and mature pupae several days prior to brood raiding. Per foundress fecundity remains constant across laboratory starting colonies established with 1, 3 and 5 foundresses. These results suggest that the selective advantage of cooperative colony foundation (pleometrosis) in this and similar species may derive directly from the ability of multiple foundresses to produce a larger brood raiding force.     

Modelling biocontrol of

The two haplotypes of Varroa destructor that have been identified as parasites of the Western honeybee (Apis mellifera L.) show disparate levels of virulence towards honeybee colonies. The Korea haplotype has been associated with severe colony mortality, whereas untreated colonies of European A. mellifera have survived long-term infestation by the Japan haplotype. The possible existence of a benign haplotype of V. destructor raises the prospect that it be used to “inoculate” colonies to provide biocontrol of the virulent haplotype. The feasibility of such a strategy was investigated using a mathematical model. Competition for resources during reproduction is known to reduce varroa mites’ reproduction rates as their infestation levels increase. Results from modelling suggested this density-dependent effect is sufficient for an established benign population to prevent the virulent population reaching destructive levels if a colony is subject to sporadic influxes of virulent mites. A colony faced with a continuous influx of mites could be protected if the proportion of virulent mites in the influx were below a threshold level (dependent on length of breeding season and intensity of influx). This condition might be achieved by “inoculating” neighbouring apiaries and controlling feral colonies in the vicinity. Decreased brood cell invasion rate by the benign haplotype decreased the threshold level. Any reproductive isolation between the benign and virulent haplotypes would cause further reproductive suppression, driving sporadic influxes of the virulent haplotype to extinction and conferring greater tolerance to a colony faced with a virulent influx. Increased colony resistance to varroa in the model was synergistic with the inoculation of colonies in the absence of reproductive isolation, but potentially antagonistic in its presence—although not to an extent that would preclude their joint use.     

Parasitism and oviposition of Melittobia acasta on Bombus terrestris: Impact of larval overwintering and host status on pupation and adult emergence

Melittobia acasta (Walker) are microhymenopteran ectoparasitoids of the pupae and prepupae of the commercially‐used pollinator bumblebee species Bombus terrestris L. The female parasitoids puncture the host cuticle with their sting and feed oozing hemolymph. This study shows that M. acasta parasitize 100% pupae and 84% prepupae of B. terrestris but are ineffective on the larvae of the bees. The female parasitoids lay a significantly higher number of eggs on pupae (67.7 ± 16.2 female^?1) compared to prepupae (20.5 ± 14.5 female^?1). The parasitoids differ in their choice for oviposition sites and fecundity on different locations of B. terrestris pupae, and they show most preference for oviposition (32%) as well as fecundity (34.9 ± 15.1 female^?1) on the petiole of the host. Larvae of the parasitoids overwinter at low temperatures but larval overwintering duration and post‐diapause rearing on original or new hosts do not affect their pupation and adult emergence. Larvae have a higher percentage of pupation (88.0–94.4%) and adult emergence (84.4–92.9%) both on the original and the new host, thus indicate that the parasitoids are highly capable of reproduction in B. terrestris colonies.     

Communal use of integumental wounds in honey bee (Apis mellifera) pupae multiply infested by the ectoparasitic mite Varroa destructor

The ectoparasitic bee mite, Varroa destructor, is highly adapted to its natural and adopted honey bee hosts, Apis cerana and Apis mellifera. Adult females perforate the integument of bee pupae in such a way that they and their progeny can feed. We examined the wounds that founder females made, and usually found one, and rarely up to three, integumental wounds on pupae of A. mellifera multiply infested by V. destructor. The punctures were mainly on the 2nd abdominal sternite of the host. These perforations are used repeatedly as feeding sites by these hemolymph-sucking mites and by their progeny. The diameter of the wounds increased during pupal development. In brood cells containing 4-5 invading female mites and their progeny, healing of the wound is delayed, normally occurring just before the imaginal moult of the bee pupa. These wounds are subject to microbial infections, and they are relevant to the evolution of behavioral traits in these parasitic mites and their relations to host bees.     

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.     

Ability to host regulate determines host choice and reproductive success in the gregarious ectoparasitoid Eulophus pennicornis (Hymenoptera: Eulophidae)

Abstract.  The age of Lacanobia oleracea (L.) in the final (sixth) larval stadium influences host choice and developmental success significantly in the gregarious ectoparasitoid Eulophus pennicornis (Nees). In choice tests, parasitoids with prior oviposition experience parasitize hosts in the second day of the sixth stadium most frequently. Parasitoid brood survival on normally-reared (i.e. fed) hosts declines monotonically with age such that mean progeny survival (egg–adult) is less than 20% for wasps developing on hosts parasitized on day 5 of the sixth stadium, as opposed to almost 50% when developing on those parasitized on day 1. Neck ligation of hosts increases the survival of wasp larvae developing on older hosts (days 4 and 5), whereas starved hosts produce progeny in similar numbers to fed hosts on most days during the final larval stadium. Hosts parasitized early in the stadium (days 1–3), although continuing to grow, do not exhibit the characteristic physical changes that non-parasitized larvae exhibit prior to pupation. However, hosts parasitized on days 4 and 5 form prepupae in appreciable numbers, particularly on day 5 where, regardless of treatment, over 80% of hosts attain this stage. Envenomated hosts behave similarly, an observation that suggests that it is the wasp's inability to arrest completely development in older hosts that is the significant factor in reducing the developmental success of the wasp. The findings are discussed in the light of the known endocrinological events in the host, and in relation to previously reported host manipulations induced by this wasp.     

Varroa management in small bites

Chelifers (Arachnida: Pseudoscorpionida), also known as pseudoscorpions, have been reported to be beneficial honeybee hive generalist pest predators for over 100 years and are occasionally noted by beekeepers in their hives. We collected chelifers within or closely associated with beehives in New Zealand. Under video observation conditions, they predated upon varroa mites while studiously ignoring bee larvae. Varroa mites reproduce at exponential rates during the spring season, and current chemical miticides rely on single treatments aiming for at least 90% control. An alternate strategy, removal of mites at a rate matching their reproductive capacity, although mathematically obvious, fails unless a suitable biological control agent is available. Our observations build on over 100 years of sporadic work to provide further evidence that chelifers show clear potential to be a suitable predator for varroa management in beehives. Approximately 25 chelifers can be expected to manage varroa populations in a single hive.     

Observations on the initiation and stimulation of oviposition of theVarroa mite

FemaleVarroa mites were collected from adult bees and were classified as swollen or not swollen. After introduction of these mites into recently sealed worker brood cells the average number of offspring of reproducing swollen mites was similar to that of naturally invaded mites, but the non-swollen mites produced a significantly lower number of offspring. This suggests that the oviposition of adult mites is stimulated by a preceding stay on adult bees. When (non-swollen) mites collected from brood cells were kept for 35 days in Eppendorf test tubes containing a larva or a pupa, their reproduction was similar to that of swollen mites.Contact of young mites, collected from brood cells, with adult bees was not essential for the initiation of oviposition. However, the number of offspring of reproducing mites, even after a third or fourth introduction into brood cells, was as a rule lower than that of mites that had been in contact with adult bees.The period of artificial introduction into sealed brood cells proved to be essential for subsequent reproduction. When introduced 48–52 h or 72–76 h after cell sealing, the mites did not produce eggs. When introduced 0–4 h after cell sealing a high percentage of the mites reproduced. Contact of the mites with a spinning larva seems necessary for initiation of oviposition.     

Fatty acid composition of the parasitic mite Varroa destructor and its host the worker prepupae of Apis mellifera

The present study analyzes the fatty acid (FA) profile of lipids isolated from Varroa destructor Anderson & Trueman, a parasitic mite of the honey bee (Apis mellifera L.), uninfected and infected worker prepupae of the Carnolian subspecies Apis mellifera carnica Pollmann, and bee bread fed to the worker brood. Significant differences are observed in the FA profiles of lipids isolated from parasites, hosts and bee bread. Parasitism by V. destructor (henceforth, varroosis) induces visible changes in the lipid profile of worker prepupae. In infected prepupae, the percentage of total saturated FAs is lower and the percentage of unsaturated FAs is higher than in uninfected insects. These differences result from significant changes in the percentages of FAs that are most abundant in the evaluated groups (i.e. C16:0, C18:1 9c, C18:2n‐6 and C18:3n‐3 FAs). In mites and in uninfected and infected prepupae, the predominant FAs are oleic acid (41.07 ± 2.26%, 42.79 ± 1.21% and 45 ± 0.20%, respectively) and palmitic acid (22.62 ± 0.87%, 39.48 ± 0.43% and 36.84 ± 0.22%, respectively). Highly significant differences in FA composition are noted between bee bread and worker brood. The results suggest specific mechanisms of FA uptake, accumulation and metabolism in the food chain of this parasitic association, beginning from the food processed by nurse bees for larval feeding, through host organisms (worker brood) to V. destructor mites.