Grooming and hygienic behaviours of the honey bee (Apis mellifera) toward the parasitic mite Varroa destructor

Despite a multitude of environmental stressors, the Varroa mite is still regarded as the greatest cause of honey bee mortality in its invaded range. Breeding honey bees that are resistant to the mite is an important area of research. This thesis aimed to gain a better understanding of the grooming and hygienic behaviours of Russian honey bees (RHB). The effect of a break in the synchrony of a mite’s life cycle on reproductive success was tested through brood inoculation experiments. Mites released by hygienic behaviour and forced to enter a new cell are less likely to lay male offspring. Through laboratory cage assays it was found that daughter mites are more susceptible to grooming behaviour. A new method of marking Varroa mites was developed which would enable a single cohort of mites to be followed after inoculation. A strong brood removal trait was noticed in RHB colonies, therefore they were tested for Varroa sensitive hygienic (VSH) behaviour. RHB demonstrated levels of VSH as high as the USDA line bred specifically for this behaviour. In addition the same QTL found to be responsible for the trait in VSH bees, was associated with VSH in RHB stock. Previous work showed that the ratio of older mites to total trapped mites (O/T) in the debris of honey bee colonies demonstrated the strongest association with colony infestation. This research showed that O/T is associated with VSH and brood removal behaviour. In addition, bees that displayed high levels of VSH in this study were also more likely to spend a longer amount of time grooming in laboratory assays. This indicates that both grooming and hygienic behaviours play important roles in the resistance of RHB stock. Their likelihood to be expressed by other stocks is discussed and recommendations for further research are provided.

Temporal changes in Varroa destructor fertility and haplotype in Brazil

The relationship between fertility and haplotype was studied in Varroa destructor mites sampled from colonies of A. mellifera carnica and Africanized Honeybees ( Apis mellifera) in Germany and Brazil respectively. Both in Germany and in Brazil, only the V. destructor Korea haplotype was found, though the Japan-Thailand haplotype was formerly thought to have been more abundant in Brazil. The fertility of Varroa mites in Brazil has increased since 1998 and is currently ( 2001) at European levels. Temporal changes in mite fertility and haplotype are not fully congruent.

Flujo génico, comportamiento defensivo y efecto de la temperatura en colonias de Apis mellífera infestadas con Varroa destructor

Tesis ( Doctor en Manejo de Recursos Naturales) U.A.N.L.

Application d’une stratégie de lutte intégrée contre le parasite Varroa destructor dans les colonies d’abeilles mellifères du Québec

Le parasite Varroa destructor provoque depuis plus de 30 ans la perte de nombreuses colonies à travers le monde. L’utilisation d’acaricides de synthèse s’est avérée inefficace au Canada et ailleurs dans le monde à la suite de la sélection de varroas résistants. Dans ce contexte, il est devenu impératif de trouver de nouveaux moyens pour contrôler cette peste apicole. Ce travail original de recherche a pour but de déterminer les paramètres fondamentaux d’une lutte intégrée contre la varroase fondée sur l’utilisation périodique de différents pesticides organiques (l’acide oxalique, l’acide formique et le thymol) associée à des seuils d’interventions. Les seuils d’intervention ont été déterminés à l’aide de régressions linéaires entre les taux de parasitisme par V. destructor et la formance zootechnique des colonies d’abeilles mellifères (production de miel et force des colonies). Un total de 154 colonies d’abeilles du Centre de recherche en sciences animales de Deschambault (CRSAD) ont été suivies de septembre 2005 à septembre 2006. Les seuils calculés et proposés à la suite de cette recherche sont de 2 varroas par jour (chute naturelle) au début mai, 10 varroas par jour à la fin juillet et de 9 varroas par jour au début septembre. L’efficacité des traitements organiques avec l’acide oxalique (AO), l’acide formique (AF) et le thymol a été vérifiée en mai (avant la première miellée) en juillet (entre deux miellées), en septembre (après la miellée et pendant le nourrissage des colonies) et en novembre (avant l’hivernage). L’acide oxalique a été appliqué en utilisant la méthode d’égouttement (4% d’AO p/v dans un sirop de sucrose 1 :1 p/v). L’acide formique a été appliquée sous forme de MiteAwayII™ (tampon commercial imbibé d’AF 65% v/v placé sur le dessus des cadres à couvain), Mitewipe (tampons Dri-Loc™ 10/15cm imbibés de 35 mL d’AF 65% v/v placés sur le dessus des cadres à couvain) ou Flash (AF 65% coulé directement sur le plateau inférieur d’une colonie, 2 mL par cadre avec abeilles). Le thymol a été appliqué sous forme d’Apiguard™ (gélose contenant 25% de thymol p/v placée sur le dessus des cadres à couvain). Les essais d’efficacité ont été réalisés de 2006 à 2008 sur un total de 170 colonies (98 appartenant au CRSAD et 72 appartenant au privé). Les résultats montrent que les traitements de printemps testés ont une faible efficacité pour le contrôle des varroas qui sont en pleine croissance durant cette période. Un traitement avec l’AF à la mi-été permet de réduire les taux de parasites sous le seuil en septembre mais il y a risque de contaminer la récolte de miel avec des résidus d’AF. Les traitements en septembre avec le MiteAwayII™ suivis par un traitement à l’acide oxalique en novembre (5 mL par égouttement entre chaque cadre avec abeilles, 4% d’AO p/v dans un sirop de sucrose 1 :1 p/v) sont les plus efficaces : ils réduisent les niveaux de varroase sous le seuil de 2 varroas par jour au printemps. Nos résultats montrent également que les traitements réalisés tôt en septembre sont plus efficaces et produisent des colonies plus fortes au printemps comparativement à un traitement réalisé un mois plus tard en octobre. En conclusion, ce travail de recherche démontre qu’il est possible de contenir le développement de la varroase dans les ruchers au Québec en utilisant une méthode de lutte intégrée basée sur une combinaison d’applications d’acaricides organiques associée à des seuils d’intervention.

Inference on microsatellite mutation processes in the invasive mite, Varroa destructor, using reversible jump Markov chain Monte Carlo

Varroa destructor is a parasitic mite of the Eastern honeybee Apis cerana. Fifty years ago, two distinct evolutionary lineages (Korean and Japanese) invaded the Western honeybee Apis mellifera. This haplo-diploid parasite species reproduces mainly through brother sister matings, a system which largely favors the fixation of new mutations. In a worldwide sample of 225 individuals from 21 locations collected on Western honeybees and analyzed at 19 microsatellite loci, a series of de novo mutations was observed. Using historical data concerning the invasion, this original biological system has been exploited to compare three mutation models with allele size constraints for microsatellite markers: stepwise (SMM) and generalized (GSM) mutation models, and a model with mutation rate increasing exponentially with microsatellite length (ESM). Posterior probabilities of the three models have been estimated for each locus individually using reversible jump Markov Chain Monte Carlo. The relative support of each model varies widely among loci, but the GSM is the only model that always receives at least 9% support, whatever the locus. The analysis also provides robust estimates of mutation parameters for each locus and of the divergence time of the two invasive lineages (67,000 generations with a 90% credibility interval of 35,000-174,000). With an average of 10 generations per year, this divergence time fits with the last post-glacial Korea Japan land separation. (c) 2005 Elsevier Inc. All rights reserved.

A importância dos mecanismos comportamentais de resistência para a dinâmica populacional de abelhas Apis mellifera e o parasita Varroa destructor

Os ácaros ectoparasitas Varroa destructor, que parasitam as abelhas tornaram-se um problema global. Embora seja pouco provável que estes ácaros, por si só, provoquem a mortalidade das colmeias, eles desempenham um importante papel como vetor de muitas doenças virais. E estas doenças são identificados como algumas das mais importantes razões para a Desordem do Colapso das Colônias. Os efeitos da infestação do V.destructor são distintas em diferentes partes do mundo. Maiores mortalidades de colônias têm sido relatadas em colônias de abelhas européias (AE) em países da Europa, Ásia e América do Norte. No entanto, este ácaro está presente no Brasil já por muitos anos e não existem relatos de perdas em colônias das abelhas africanizadas (AA). Estudos realizados no México mostraram que alguns comportamentos de resistência ao ácaro Varroa - especialmente o grooming e o comportamento higiênico - são diferentes em cada uma das subespécie. Poderiam então esses mecanismos explicar por que as abelhas africanizadas são menos suscetíveis à Desordem do Colapso das Colônias? A fim de responder a esta pergunta, propomos um modelo matemático baseado em equações diferenciais, com o objetivo de analisar o papel desses mecanismos de resistência na saúde geral da colônia e na capacidade da colônia para enfrentar desafios ambientais.

Produtos naturais para o controle do ácaro Varroa destructor em abelhas africanizadas

O objetivo deste trabalho foi avaliar os efeitos do ácido oxálico e de óleos essenciais de plantas no controle da infestação pelo ácaro Varroa destructor em colônias de Apis mellifera africanizadas. O experimento foi realizado em delineamento inteiramente casualizado, em 30 colônias, com seis tratamentos e cinco repetições. As colmeias foram tratadas com óleos essenciais de arruda (Ruta graveolens), eucalipto (Eucalyptus spp.) e hortelã (Mentha piperita), além de timol, ácido oxálico e do tratamento controle sem aplicação de produtos. Foram avaliadas a mortalidade de varroas e as taxas de mortalidade de crias e de infestação de varroas em crias e em abelhas adultas, antes e depois da aplicação de cada produto. O ácido oxálico e os óleos de arruda, timol, eucalipto e de hortelã reduziram a mortalidade de crias parasitadas pelo ácaro em 92,1, 83,3, 81,7, 86,4 e 81,3%, respectivamente. O tratamento com ácido oxálico reduziu em 87,4% a infestação de varroas em abelhas adultas. O uso desses produtos é eficiente na redução da mortalidade de crias de A. mellifera parasitadas por V. destructor.

Analysis of Mortality in Africanized Honey Bee Colonies with High Levels of Infestation by Varroa destructor

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Effects of Natural Infestations of the Mite Varroa destructor on the Development of Africanized Honeybee Workers (Apis mellifera)

The mite Varroa destructor (Anderson & Treuman 2000) has caused extensive damage to beekeeping worldwide. In Brazil, weather conditions and the strains of bees do not provide ideal conditions for mite parasitism, which is reflected in the low number of deaths of colonies caused by varroatosis well as the stability of infestation levels. The aim of this study was to evaluate the damage caused by the mite infestation in hives maintained in natural conditions. For this purpose the number of mites per bee was calculated and used to quantify the level of infestation in each colony. To record the mortality rates of parasitized bees during development daily checks were performed. The data were analyzed by G test of independence and a Test of Proportions. The results indicate that the rate of mortality of pupae and larvae was proportional to the degree of infestation in each colony, and all colonies showed mortality rates significantly higher than the control rate. A significant interaction among death rates recorded between the third and fourth days of larval life and the total death of larvae was found (G Test = 50.22; P < 0.0001). So, it can be concluded that bee inbreeding contributed significantly to the increase of the larval rate of mortality. In Africanized honeybee colonies infested by the mite Varroa destructor mortality rates in conditions of natural infestation varied from 6.65 to 9.89% in pupae (<(x)over bar>= 8.78%) and from 6.13 to 13.48% in larvae ((x) over bar = 9.91%), against 3.85% and 3.74% in the control colony, respectively. Therefore, in the infested colonies the average rates of mortality caused by the harmful effects of the mite were, respectively, 2.28 times and 2.65 times greater in those two developmental stages.

Efeito da infestação do ácaro Varroa destructor (Anderson e Treuman, 2000) (Arachnida : Acari : Varroidae) no desenvolvimento de abelhas africanizadas Apis mellifera (Linnaeus, 1758) (Hymenoptera: Apidae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Produtos naturais no controle do ácaro Varroa destructor em abelhas Apis mellifera L. (africanizadas)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Winter Losses of Honeybee Colonies (Hymenoptera: Apidae): The Role of Infestations With Aethina tumida (Coleoptera: Nitidulidae) and Varroa destructor (Parasitiformes: Varroidae)

Multiple infections of managed honeybee, Apis mellifera, colonies are inevitable due to the ubiquitous ectoparasitic mite Varroa destructor and might be an underlying cause of winter losses. Here we investigated the role of adult small hive beetles, Aethina tumida, alone and in combination with V. destructor for winter losses and for infections with the microsporidian endoparasite Nosema ceranae. We found no significant influence of A. tumida and V destructor alone or in combination on the numbers of N. ceranae spores. Likewise, A. tumida alone had no significant effects on winter losses, which is most likely due to the observed high winter mortality of the adult beetles. Therefore, our data suggest that A. tumida is unlikely to contribute to losses of overwintering honeybee colonies. However, high losses occurred in all groups highly infested with V. destructor, supporting the central role of the mite for colony losses.

Honey Bee Apis mellifera Parasites in the Absence of Nosema ceranae Fungi and Varroa destructor Mites

Few areas of the world have western honey bee (Apis mellifera) colonies that are free of invasive parasites Nosema ceranae (fungi) and Varroa destructor (mites). Particularly detrimental is V. destructor; in addition to feeding on host haemolymph, these mites are important vectors of several viruses that are further implicated as contributors to honey bee mortality around the world. Thus, the biogeography and attendant consequences of viral communities in the absence of V. destructor are of significant interest. The island of Newfoundland, Province of Newfoundland and Labrador, Canada, is free of V. destructor; the absence of N. ceranae has not been confirmed. Of 55 Newfoundland colonies inspected visually for their strength and six signs of disease, only K-wing had prevalence above 5% (40/55 colonies = 72.7%). Similar to an earlier study, screenings again confirmed the absence of V. destructor, small hive beetles Aethina tumida (Murray), tracheal mites Acarapis woodi (Rennie), and Tropilaelaps spp. ectoparasitic mites. Of a subset of 23 colonies screened molecularly for viruses, none had Israeli acute paralysis virus, Kashmir bee virus, or sacbrood virus. Sixteen of 23 colonies (70.0%) were positive for black queen cell virus, and 21 (91.3%) had some evidence for deformed wing virus. No N. ceranae was detected in molecular screens of 55 colonies, although it is possible extremely low intensity infections exist; the more familiar N. apis was found in 53 colonies (96.4%). Under these conditions, K-wing was associated (positively) with colony strength; however, viruses and N. apis were not. Furthermore, black queen cell virus was positively and negatively associated with K-wing and deformed wing virus, respectively. Newfoundland honey bee colonies are thus free of several invasive parasites that plague operations in other parts of the world, and they provide a unique research arena to study independent pathology of the parasites that are present.

Genome characterization, prevalence and distribution of a Macula-like virus from Apis mellifera and Varroa destructor

Around 14 distinct virus species-complexes have been detected in honeybees, each with one or more strains or sub-species. Here we present the initial characterization of an entirely new virus species-complex discovered in honeybee (Apis mellifera L.) and varroa mite (Varroa destructor) samples from Europe and the USA. The virus has a naturally poly-adenylated RNA genome of about 6500 nucleotides with a genome organization and sequence similar to the Tymoviridae (Tymovirales; Tymoviridae), a predominantly plant-infecting virus family. Literature and laboratory analyses indicated that the virus had not previously been described. The virus is very common in French apiaries, mirroring the results from an extensive Belgian survey, but could not be detected in equally-extensive Swedish and Norwegian bee disease surveys. The virus appears to be closely linked to varroa, with the highest prevalence found in varroa samples and a clear seasonal distribution peaking in autumn, coinciding with the natural varroa population development. Sub-genomic RNA analyses show that bees are definite hosts, while varroa is a possible host and likely vector. The tentative name of Bee Macula-like virus (BeeMLV) is therefore proposed. A second, distantly related Tymoviridae-like virus was also discovered in varroa transcriptomes, tentatively named Varroa Tymo-like virus (VTLV).