No abstract

Very rapidly after Varroa destructor invaded apiaries of Apis mellifera, the devastating effect of this mite prompted an active research effort to understand and control this parasite. Over a few decades, varroa has spread to most countries exploiting A. mellifera. As a consequence, a large number of teams have worked with this organism, developing a diversity of research methods. Often different approaches have been followed to achieve the same goal. The diversity of methods made the results difficult to compare, thus hindering our understanding of this parasite. In this paper, we provide easy to use protocols for the collection, identification, diagnosis, rearing, breeding, marking and measurement of infestation rates and fertility of V. destructor. We also describe experimental protocols to study orientation and feeding of the mite, to infest colonies or cells and measure the mite’s susceptibility to acaricides. Where relevant, we describe which mite should be used for bioassays since their behaviour is influenced by their physiological state. We also give a method to determine the damage threshold above which varroa damages colonies. This tool is fundamental to be able to implement integrated control concepts. We have described pros and cons for all methods for the user to know which method to use under which circumstances. These methods could be embraced as standards by the community when designing and performing research on V. destructor

Honey bees are increasingly important in the pollination of crops and wild plants. Recent reports of the weakening and periodical high losses of managed honey bee colonies have alarmed beekeeper, farmers and scientists. Infestations with the ectoparasitic mite Varroa destructor in combination with its associated viruses have been identified as a crucial driver of these health problems. Although yearly treatments are required to prevent collapses of honey bee colonies, the number of effective acaricides is small and no new active compounds have been registered in the past 25 years. RNAi-based methods were proposed recently as a promising new tool. However, the application of these methods according to published protocols has led to a surprising discovery. Here, we show that the lithium chloride that was used to precipitate RNA and other lithium compounds is highly effective at killing Varroa mites when fed to host bees at low millimolar concentrations. Experiments with caged bees and brood-free artificial swarms consisting of a queen and several thousand bees clearly demonstrate the potential of lithium as miticidal agent with good tolerability in worker bees providing a promising basis for the development of an effective and easy-to-apply control method for mite treatment.

During a reproductive cycle, not all daughter mites of Varroa destructor mate and thus leave the brood cells as virgins. Here, we show that virgin mites are present within both the phoretic (10%) and reproductive (8%) mite population. Most (n = 29 of n = 33) of these encountered virgins laid unfertilized (= male) eggs, and some (n = 10) mated later on with their own son. These findings were verified by tests with artificially reared virgin mites. Obviously, mating is not a prerequisite for Varroa reproduction. However, due to the small number of reproductive cycles, the contribution of virgins to the Varroa population is regarded as low. This study also confirms conclusively that sex of V. destructor is determined via arrhenotokous parthenogenesis and not-as previously assumed-via pseudo-arrhenotoky. Furthermore, reproductive parameters of naturally invaded and artificially introduced Varroa females were compared, and artificial infestation was reconfirmed as a suitable method.

Mating of Varroa destructor takes place inside the sealed honey bee brood cell. During copulation, male mites transfer the spermatozoa into the genital openings of the females. Before the fertilization of female germ cells, the transferred spermatozoa have to pass through a final maturation process inside the genital tract of the female, the so-called capacitation. We here describe for the first time the morphological changes and chronological sequence of spermatozoa capacitation within female V. destructor. We have defined seven distinct stages of spermatozoa during the process of capacitation and have shown that it takes about 5 days from mating to the occurrence of spermatozoa ready for fertilization. This might explain the results of an additional experiment where we could show that freshly mated daughter mites need a phoretic phase on bees before their first reproduction cycle. The transfer of non-capacitated spermatozoa from male V. destructor and the resulting long capacitation period within the female mites seems to be a consequence of an adaptive pressure for the male mites to inseminate several daughter mites within the short time span inside the sealed honey bee brood cell.