Relations entre le parasite Varroa destructor et son hôte l'abeille domestique : étude des facteurs impactant le déroulement du cycle de développement du parasite

Varroa destructor, principal ectoparasite de l'abeille, est l'un des responsables majeurs des phénomènes de perte de colonies observés partout dans le monde. Depuis son changement d'hôte et l'infestation de colonies d'Apis mellifera dans les années 1970, des traitements essentiellement chimiques ont été proposés pour limiter l'infestation. Pourtant ces dernières années, ces traitements ont fait l'objet d'études critiques à la fois à cause de la résistance émergente du parasite aux différents acaricides et du fait de la sensibilité des abeilles à l'accumulation de produits chimiques dans les différentes matrices de la ruche. De nouveaux traitements alternatifs contre la varroose sont donc nécessaires et très attendus afin d'enrayer rapidement les effets délétères du parasitisme acarien. Cependant, la recherche de ces nouvelles cibles thérapeutiques est limitée par l'absence d'un système d'élevage synchronisé de l'hôte et de son parasite en laboratoire. L'objectif technique de ce travail de thèse était donc de pallier à ce manque en développant une méthode pour coupler l'élevage de larves d'abeilles in vitro avec le transfert à la fois de la phase phorétique et de la phase de reproduction du parasite (respectivement sur les adultes et sur les pupes) dans des conditions de laboratoire. Le système mis au point lors de ces travaux de thèse a permis une étude plus approfondie de l'impact de facteurs variables chez l'hôte sur plusieurs points clés du cycle parasitaire. Les effets de la nutrition larvaire chez l'abeille, la longueur de la phase phorétique, le type d'abeilles adultes sur lequel Varroa reste au cours de cette phase ou encore l'âge des stades larvaires ont ainsi été étudiés. L'analyse des impacts potentiels de ces facteurs a porté principalement sur certains paramètres de la reproduction, du comportement et de la virulence de V. destructor. Les résultats obtenus ont montré que la nutrition larvaire, ou plus précisément la supplémentation de la nourriture larvaire avec des pollens monofloraux et polyfloraux, a un faible impact sur la reproduction du parasite, mais présente une tendance à influencer le poids de l'abeille ainsi qu'à potentiellement augmenter la tolérance de l'abeille à certains des symptômes de la varroose, tels que la perte de poids. La reproduction du parasite n'a pas non plus été affectée par les variations induites dans la phase phorétique. Ni la fertilité du parasite, ni l'expression des deux gènes de la vitellogénine n'ont fluctué lorsque la durée ou l'âge des adultes parasités pendant la phase phorétique variaient. En revanche, la durée du séjour sur les abeilles adultes avant la reproduction est corrélée positivement à la charge virale chez le parasite, ce qui explique pourquoi les varroas ayant vécu une longue phase phorétique induisent plus fréquemment des malformations. Enfin, l'âge des stades juvéniles des abeilles sont tous attrayants pour Varroa destructor, bien que certaines préférences aient été mises en évidence dans notre étude. Si l'attraction pour le 5ème stade larvaire et les abeilles nouvellement écloses est déjà connu, l'attractivité des nymphes aux yeux blancs est un phénomène nouveau qui pourrait être d'importance majeure pour la compréhension du cycle parasitaire. Des analyses approfondies pour isoler les signaux responsables de cette attraction restent nécessaires. En conclusion, les travaux de cette thèse offrent une méthode d'élevage innovante et ouvrent ainsi, pour l'ensemble de la communauté apicole, de nouvelles avenues de recherche passionnantes afin de développer nos connaissances à la fois sur l'abeille, sur le parasite et sur les virus qu'il transporte. Ce travail pourrait permettre d'identifier de nouvelles cibles thérapeutiques pour la conception de traitements alternatifs ciblant spécifiquement le parasite sans nuire à l'abeille.Varroa destructor is the principal acarian parasite of the honey bee, involved in the phenomenon of colony loss which has been reported worldwide. Since its host shift from Apis cerana to Apis mellifera in the 1970's, chemical treatments have been set up to limit infestation. For many years however, these treatments have been the subject of critical studies both because of the resistance of the parasite to acaricides and of the sensitivity of bees to chemical accumulation in hive matrices. New alternative ways of varroatosis treatment are therefore urgently needed to limit the deleterious effects of the acarian parasitism. However, the search for these alternative therapeutic targets is limited by the lack of a laboratory system to transfer and synchronize the cycles of V. destructor and the honey bee under controlled conditions. The methodological objective of this Phd work was to overcome this issue by developing a method to couple the rearing of honey bee larvae in vitro to the transfer of both the parasite phoretic and reproductive phases (respectively on adults and on pupae) in laboratory conditions. Our system allowed further investigation regarding the impact of variable parameters in the host on several key points of the parasite cycle. The effects of several factors -such as the bee larval nutrition, the length of the phoretic phase, the type of adult bees on which Varroa stays during this phase or the age of pupal development- were studied. The potential impact in the parasite cycle focused mainly on reproductive, behavioural and virulence related parameters. The results obtained from our experiments showed that larval nutrition, tested via the supplementation of the larval food with pollen, had no impact on the reproduction of the parasite but a tendency to influence the weight of the bee, potentially increasing the bee tolerance to some of the symptoms of varroatosis, such as the weight loss. The reproduction of Varroa was not impacted by variations induced in the phoretic phase either. Neither the fertility of the parasite nor the expression of its two vitellogenin genes were impacted by the duration or the age of adults parasitized during the phoretic phase. On the other hand, the length of the stay on adult bees before reproduction positively correlates with the viral loads in the parasite, which explains why varroas experiencing a long phoretic phase induce malformation more frequently in parasitized pupae. Finally, the age of juvenile stages of the honey bees are all attractive to Varroa destructor, although some preferences have been highlighted in our study. If the attraction for 5th instar larvae and newly emerged bees have already been described, the attractions towards the white eyed pupae is a new phenomenon that could be of major importance in the understanding of the parasite cycle. Further analysis to isolate and shed light on the attraction signals involved in the process is needed. Altogether, this thesis has supplied, for the whole scientific beekeeping community, new exciting research avenues and methodologies to develop our knowledge about the bee, the parasite and the viruses it carries. It might allow us, in the future, to design alternative treatment specifically targeting the parasite without injuring the honey bee

Interactions between the ectoparasite Varroa destructor and its host the European honey bee Apis mellifera : study of the factors impavting the parasite developmental cycle

Varroa destructor, principal ectoparasite de l'abeille, est l'un des responsables majeurs des phénomènes de perte de colonies observés partout dans le monde. Depuis son changement d'hôte et l'infestation de colonies d'Apis mellifera dans les années 1970, des traitements essentiellement chimiques ont été proposés pour limiter l'infestation. Pourtant ces dernières années, ces traitements ont fait l'objet d'études critiques à la fois à cause de la résistance émergente du parasite aux différents acaricides et du fait de la sensibilité des abeilles à l'accumulation de produits chimiques dans les différentes matrices de la ruche. De nouveaux traitements alternatifs contre la varroose sont donc nécessaires et très attendus afin d'enrayer rapidement les effets délétères du parasitisme acarien. Cependant, la recherche de ces nouvelles cibles thérapeutiques est limitée par l'absence d'un système d'élevage synchronisé de l'hôte et de son parasite en laboratoire. L'objectif technique de ce travail de thèse était donc de pallier à ce manque en développant une méthode pour coupler l'élevage de larves d'abeilles in vitro avec le transfert à la fois de la phase phorétique et de la phase de reproduction du parasite (respectivement sur les adultes et sur les pupes) dans des conditions de laboratoire. Le système mis au point lors de ces travaux de thèse a permis une étude plus approfondie de l'impact de facteurs variables chez l'hôte sur plusieurs points clés du cycle parasitaire. Les effets de la nutrition larvaire chez l'abeille, la longueur de la phase phorétique, le type d'abeilles adultes sur lequel Varroa reste au cours de cette phase ou encore l'âge des stades larvaires ont ainsi été étudiés. L'analyse des impacts potentiels de ces facteurs a porté principalement sur certains paramètres de la reproduction, du comportement et de la virulence de V. destructor. Les résultats obtenus ont montré que la nutrition larvaire, ou plus précisément la supplémentation de la nourriture larvaire avec des pollens monofloraux et polyfloraux, a un faible impact sur la reproduction du parasite, mais présente une tendance à influencer le poids de l'abeille ainsi qu'à potentiellement augmenter la tolérance de l'abeille à certains des symptômes de la varroose, tels que la perte de poids. La reproduction du parasite n'a pas non plus été affectée par les variations induites dans la phase phorétique. Ni la fertilité du parasite, ni l'expression des deux gènes de la vitellogénine n'ont fluctué lorsque la durée ou l'âge des adultes parasités pendant la phase phorétique variaient. En revanche, la durée du séjour sur les abeilles adultes avant la reproduction est corrélée positivement à la charge virale chez le parasite, ce qui explique pourquoi les varroas ayant vécu une longue phase phorétique induisent plus fréquemment des malformations. Enfin, l'âge des stades juvéniles des abeilles sont tous attrayants pour Varroa destructor, bien que certaines préférences aient été mises en évidence dans notre étude. Si l'attraction pour le 5ème stade larvaire et les abeilles nouvellement écloses est déjà connu, l'attractivité des nymphes aux yeux blancs est un phénomène nouveau qui pourrait être d'importance majeure pour la compréhension du cycle parasitaire. Des analyses approfondies pour isoler les signaux responsables de cette attraction restent nécessaires. En conclusion, les travaux de cette thèse offrent une méthode d'élevage innovante et ouvrent ainsi, pour l'ensemble de la communauté apicole, de nouvelles avenues de recherche passionnantes afin de développer nos connaissances à la fois sur l'abeille, sur le parasite et sur les virus qu'il transporte. Ce travail pourrait permettre d'identifier de nouvelles cibles thérapeutiques pour la conception de traitements alternatifs ciblant spécifiquement le parasite sans nuire à l'abeille.Varroa destructor is the principal acarian parasite of the honey bee, involved in the phenomenon of colony loss which has been reported worldwide. Since its host shift from Apis cerana to Apis mellifera in the 1970's, chemical treatments have been set up to limit infestation. For many years however, these treatments have been the subject of critical studies both because of the resistance of the parasite to acaricides and of the sensitivity of bees to chemical accumulation in hive matrices. New alternative ways of varroatosis treatment are therefore urgently needed to limit the deleterious effects of the acarian parasitism. However, the search for these alternative therapeutic targets is limited by the lack of a laboratory system to transfer and synchronize the cycles of V. destructor and the honey bee under controlled conditions. The methodological objective of this Phd work was to overcome this issue by developing a method to couple the rearing of honey bee larvae in vitro to the transfer of both the parasite phoretic and reproductive phases (respectively on adults and on pupae) in laboratory conditions. Our system allowed further investigation regarding the impact of variable parameters in the host on several key points of the parasite cycle. The effects of several factors -such as the bee larval nutrition, the length of the phoretic phase, the type of adult bees on which Varroa stays during this phase or the age of pupal development- were studied. The potential impact in the parasite cycle focused mainly on reproductive, behavioural and virulence related parameters. The results obtained from our experiments showed that larval nutrition, tested via the supplementation of the larval food with pollen, had no impact on the reproduction of the parasite but a tendency to influence the weight of the bee, potentially increasing the bee tolerance to some of the symptoms of varroatosis, such as the weight loss. The reproduction of Varroa was not impacted by variations induced in the phoretic phase either. Neither the fertility of the parasite nor the expression of its two vitellogenin genes were impacted by the duration or the age of adults parasitized during the phoretic phase. On the other hand, the length of the stay on adult bees before reproduction positively correlates with the viral loads in the parasite, which explains why varroas experiencing a long phoretic phase induce malformation more frequently in parasitized pupae. Finally, the age of juvenile stages of the honey bees are all attractive to Varroa destructor, although some preferences have been highlighted in our study. If the attraction for 5th instar larvae and newly emerged bees have already been described, the attractions towards the white eyed pupae is a new phenomenon that could be of major importance in the understanding of the parasite cycle. Further analysis to isolate and shed light on the attraction signals involved in the process is needed. Altogether, this thesis has supplied, for the whole scientific beekeeping community, new exciting research avenues and methodologies to develop our knowledge about the bee, the parasite and the viruses it carries. It might allow us, in the future, to design alternative treatment specifically targeting the parasite without injuring the honey bee

Varroa destructor rearing in laboratory conditions: importance of foundress survival in doubly infested cells and reproduction of laboratory-born females

International audienceA considerable part of the knowledge about the honey bee parasite Varroa destructor emerged from rearing protocols in semi-natural or laboratory conditions, yet a durable protocol over several generations of mites is still lacking. The development of such multigenerational rearing relies on the emergence of a sufficient number of new fertile females in the first generation of V. destructor. The optimization of the parasite’s reproductive success in laboratory conditions thus represents an important prerequisite. The number of foundress mites in a cell is known to impact the probability of male survival and thus the number of mated daughters. We therefore investigated the effect of the degree of bee larvae infestation under laboratory conditions. The results showed that the probability of finding at least one foundress alive at the end of the rearing was significantly higher in doubly infested cells. This leads to the improvement of the reproductive parameters and more specifically of the number of daughters per mite. In doubly infested cells with one dead foundress, the presence of a surviving female would in fact allow both its descendants and those of the dead mite to complete their development. The mated daughters from this system were used in a subsequent experiment to test their ability to complete their reproductive cycle in laboratory conditions, from the perspective of developing a multigenerational rearing. The reproduction and development of the offspring measured were similar to those of the first generation. However, many of the females from the second generation died before the completion of their first reproductive cycle. We suggest that these females are fertile but might lack the energy necessary to survive throughout reproduction. The results from our bioassay could constitute a basis for the development of a durable V. destructor laboratory rearing and for the improvement of our understanding of the parasite’s reproductive cycle

Transmission of deformed wing virus between Varroa destructor foundresses, mite offspring and infested honey bees

International audienceBackground: Varroa destructor is the major ectoparasite of the western honey bee (Apis mellifera). Through both its parasitic life-cycle and its role as a vector of viral pathogens, it can cause major damage to honey bee colonies. The deformed wing virus (DWV) is the most common virus transmitted by this ectoparasite, and the mite is correlated to increased viral prevalence and viral loads in infested colonies. DWV variants A and B (DWV-A and DWV-B, respectively) are the two major DWV variants, and they differ both in their virulence and transmission dynamics. Methods: We studied the transmission of DWV between bees, parasitic mites and their offspring by quantifying DWV loads in bees and mites collected in in vitro and in situ environments. In vitro, we artificially transmitted DWV-A to mites and quantified both DWV-A and DWV-B in mites and bees. In situ, we measured the natural presence of DWV-B in bees, mites and mites' offspring. Results: Bee and mite viral loads were correlated, and mites carrying both variants were associated with higher mortality of the infected host. Mite infestation increased the DWV-B loads and decreased the DWV-A loads in our laboratory conditions. In situ, viral quantification in the mite offspring showed that, after an initially non-infected egg stage, the DWV-B loads were more closely correlated with the foundress (mother) mites than with the bee hosts. Conclusions: The association between mites and DWV-B was highlighted in this study. The parasitic history of a mite directly impacts its DWV infection potential during the rest of its life-cycle (in terms of variant and viral loads). Regarding the mite's progeny, we hypothesize that the route of contamination is likely through the feeding site rather than by vertical transmission, although further studies are needed to confirm this hypothesis

Varroa destructor from the Laboratory to the Field: Control, Biocontrol and IPM Perspectives—A Review

Varroa destructor is a real challenger for beekeepers and scientists: fragile out of the hive, tenacious inside a bee colony. From all the research done on the topic, we have learned that a better understanding of this organism in its relationship with the bee but also for itself is necessary. Its biology relies mostly on semiochemicals for reproduction, nutrition, or orientation. Many treatments have been developed over the years based on hard or soft acaricides or even on biocontrol techniques. To date, no real sustainable solution exists to reduce the pressure of the mite without creating resistances or harming honeybees. Consequently, the development of alternative disruptive tools against the parasitic life cycle remains open. It requires the combination of both laboratory and field results through a holistic approach based on health biomarkers. Here, we advocate for a more integrative vision of V. destructor research, where in vitro and field studies are more systematically compared and compiled. Therefore, after a brief state-of-the-art about the mite’s life cycle, we discuss what has been done and what can be done from the laboratory to the field against V. destructor through an integrative approach

Effect of pollen extract supplementation on the varroatosis tolerance of honey bee (Apis mellifera) larvae reared in vitro

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Standard Methods for Dissection of Varroa destructor Females

Varroa destructor (Anderson and Trueman) is known as a major pest of Apis mellifera L, especially in the Northern Hemisphere where its effects can be deleterious. As an obligate parasite, this mite relies entirely on its host to reproduce and complete its cycle. Studies focusing on isolated organs are needed to better comprehend this organism. To conduct such targeted molecular or physiological studies, the dissection of V. destructor mites is crucial as it allows the extraction of specific organs. Here, we propose a technical article showing detailed steps of females V. destructor dissection, illustrated with pictures and videos. These illustrated guidelines will represent a helpful tool to go further in V. destructor research

Dispersal in the inquiline social parasite ant Plagiolepis xene.

Inquiline ants are highly specialised social parasites. They usually do not produce their own worker caste but instead use the worker force of the host ant colony to ensure the rearing of their sexual progeny. Several barriers are expected to severely limit their migration, and the mechanism allowing them to disperse remains largely enigmatic. Here, we tested two hypotheses to account for the low level of infestation of inquiline parasites, in populations of the parasite ant Plagiolepis xene and its host P. pygmaea: (1) the establishment of a new P. xene colony is such a rare event that a single colonisation should be expected per population, and (2) once a P. xene colony is established in one location, it has very little chance to succeed in infecting a neighbour genetically unrelated colony. We sampled nests from both species along four separate transects, and genotyped host and parasite individuals at eight polymorphic microsatellite loci. Our genetic data contradict both hypotheses: multiple colonisation events were recorded in all four transects sampled and, in at least one case, P. xene has successfully migrated from one host colony of P. pygmaea to a spatially close unrelated nest. This shows that the dispersion capacity of the social parasite is sufficiently effective to ensure its long-term survival. © 2014 International Union for the Study of Social Insects (IUSSI).SCOPUS: ar.jinfo:eu-repo/semantics/publishe