Marina D. Meixner

The German bee monitoring project: a long term study to understand periodically high winter losses of honey bee colonies

The Western honey bee, Apis mellifera, is the most important animal pollinator in agriculture worldwide providing more than 90% of the commercial pollination services. Due to the development in agriculture the demands for honey bee pollination are steadily increasing stressing the pollination capacity of the global managed honey bee population. Hence, the long-term decline of managed honey bee hives in Europe and North-America is of great concern and stimulated intensive research into the possible factors presumably causing honey bee colony collapse. We here present a four-year study involving more than 1200 bee colonies from about 120 apiaries which were monitored for the entire study period. Bee samples were collected twice a year to analyze various pathogenic factors including the ectoparasitic mite Varroa destructor, fungi (Nosema spec., Ascosphaera apis), the bacterium Paenibacillus larvae, and several viruses. Data on environmental factors, beekeeping management practice, and pesticides were also collected. All data were statistically analyzed in respect to the overwintering mortality of the colonies. We can demonstrate for several factors that they are significantly related to the observed winter losses of the monitored honey bee colonies: (i) high varroa infestation level, (ii) infection with deformed wing virus (DWV) and acute bee paralysis virus (ABPV) in autumn, (iii) queen age, and (iv) weakness of the colonies in autumn. No effects could be observed for Nosema spec. or pesticides. The implications of these findings will be discussed.ZusammenfassungDie Honigbiene Apis mellifera ist weltweit der wichtigste Bestäuber in der Landwirtschaft und nach aktuellen Schätzungen wird der globale Bedarf an kommerzieller Bestäubung weiter steigen. Dadurch stellt der seit Jahren zu beobachtende stetige Rückgang der Bienenvölker in Nord-Amerika und Europa ein ernsthaftes Problem für die Landwirtschaft dar. Für die Abnahme der Bienenvölker werden neben wirtschaftlichen Faktoren vor allem periodisch auftretende Völkerverluste verantwortlich gemacht, für die aber eine eindeutige Ursachenanalyse bisher fehlt.Zur Ursachenaufklärung von Winterverlusten führten wir von 2004 bis 2009 ein Monitoringprojekt durch, in dem mehr als 1200 Bienenvölker auf 125 über ganz Deutschland verteilten Bienenständen (Abb. 1) kontinuierlich beprobt und kontrolliert wurden. Die beteiligten „Monitoringimker“ stellten hierfür 10 ihrer Völker zur Verfügung und lieferten Daten zu Honigerträgen, Wanderungen und Ablegerbildung. Mitarbeiter der Bieneninstitute nahmen zweimal im Jahr Bienenproben für Krankheitsuntersuchungen (Nosema spec, Varroa destructor, 4 verschiedene Bienenviren) sowie Bienenbrotproben für Rückstandsuntersuchungen. Die Stärke der Bienenvölker wurde bei der Ein- und Auswinterung bestimmt; als „Überwinterungsverlust“ wurden Völker definiert, die tot waren bzw. nicht genug Bienen für eine erfolgreiche Frühjahrsentwicklung aufwiesen.Die Winterverluste schwankten zwischen 3,5 % und 15,2 % (Abb. 3) mit ungleicher Verteilung innerhalb der beteiligten Imker (Abb. 4). Für die Ursachenanalyse wurden die überlebenden mit den zusammengebrochenen Völkern verglichen. Dabei zeigten sich die größten und hochsignifikanten (P < 0,000001, U-Test) Unterschiede beim Varroabefall der Bienen im Oktober (Tab. III, Abb. 5). Ebenfalls hochsignifikante Unterschiede ergaben sich für die Bienenviren DWV (P < 0,00001) und APBV (P < 0,0039), nicht jedoch für KBV, SBV und den Nosemabefall (Tab. V). Erstaunlicherweise waren Völker mit jungen Königinnen signifikant seltener von Winterverlusten betroffen als mit älteren Königinnen (Tab. VI), während z. B. Beutenmaterial oder Rähmchenmaß keine Rolle spielten.Bei den insgesamt in drei Jahren auf Pestizidrückstände untersuchten 215 Bienenbrotproben wurden insgesamt über 50 Wirkstoffe (von 256) nachgewiesen, die meisten im Spurenbereich. Häufig wurden mehrere Wirkstoffe gefunden und nur etwas mehr als 20 % der Proben waren frei von messbaren Rückständen (Tab. VII). Neonikotinoide wurden nur in einer einzigen Probe nachgewiesen. Es konnte keine Korrelation von Rückstandswerten mit Winterverlusten festgestellt werden. Es gab auch keinen Zusammenhang zwischen der Überwinterung von Bienenvölkern und dem Umfang des zuvor eingetragenen Rapshonigs (Abb. 6).Unser Projekt zeigt, dass der Varroabefall im Herbst (zusammen mit den assoziierten Sekundärinfektionen) eine Hauptursache für Überwinterungsverluste darstellt. Eine konsequente Varroabehandlung und starke Bienenvölker mit jungen Königinnen sind daher die wichtigste Empfehlung, um Winterverlusten vorzubeugen. Ein zusätzlicher Einfluss der übrigen Faktoren kann nicht ausgeschlossen werden, hierfür sind aber modifizierte Versuchsansätze notwendig.

The neonicotinoids thiacloprid, imidacloprid, and clothianidin affect the immunocompetence of honey bees (Apis mellifera L.)

A strong immune defense is vital for honey bee health and colony survival. This defense can be weakened by environmental factors that may render honey bees more vulnerable to parasites and pathogens. Honey bees are frequently exposed to neonicotinoid pesticides, which are being discussed as one of the stress factors that may lead to colony failure. We investigated the sublethal effects of the neonicotinoids thiacloprid, imidacloprid, and clothianidin on individual immunity, by studying three major aspects of immunocompetence in worker bees: total hemocyte number, encapsulation response, and antimicrobial activity of the hemolymph. In laboratory experiments, we found a strong impact of all three neonicotinoids. Thiacloprid (24h oral exposure, 200 μg/l or 2000 μg/l) and imidacloprid (1 μg/l or 10 μg/l) reduced hemocyte density, encapsulation response, and antimicrobial activity even at field realistic concentrations. Clothianidin had an effect on these immune parameters only at higher than field realistic concentrations (50-200 μg/l). These results suggest that neonicotinoids affect the individual immunocompetence of honey bees, possibly leading to an impaired disease resistance capacity.

Standard methods for characterising subspecies and ecotypes of Apis mellifera

Summary The natural diversity of honey bees in Europe is eroding fast. A multitude of reasons lead to a loss of both genetic diversity and specific adaptations to local conditions. To preserve locally adapted bees through breeding efforts and to maintain regional strains in conservation areas, these valuable populations need to be identified. In this paper, we give an overview of methods that are currently available and used for recognition of honey bee subspecies and ecotypes, or that can be utilised to verify the genetic origin of colonies for breeding purposes. Beyond summarising details of morphometric, allozyme and DNA methods currently in use, we report recommendations with regard to strategies for sampling, and suggest methods for statistical data analysis. In particular, we emphasise the importance of reference data and consistency of methods between laboratories to yield comparable results.

Conserving diversity and vitality for honey bee breeding.

Summary Beekeepers in Europe, North America and other parts of the world have repeatedly been afflicted by elevated and sometimes unexplained colony losses. Multiple factors have been considered in connection with increased winter losses. In addition to national programmes investigating possible causes for increased honey bee mortality, scientists collaborate at an international level on different aspects of bee health within the COLOSS network. Within this network, Working Group 4 explores aspects of genetic diversity in relation to the vitality and health of honey bee populations. In this paper, we briefly review the genetic diversity of honey bees in Europe, discuss the effects of beekeeping and selective breeding on honey bee populations under the aspect of genetic diversity and bee health, and review the current status of EU legislation with respect to protection of native bee populations. We introduce and discuss recent approaches in honey bee selective breeding to improve disease resistance by introducing traits related to colony vitality. Finally, we present the aims of WG4 within the COLOSS network and briefly introduce our experimental approach.

The Mediterranean fruit fly in California: evidence for multiple introductions and persistent populations based on microsatellite and mitochondrial DNA variability

Microsatellite and mitochondrial DNA (mtDNA) variability data were used to study outbreaks of Mediterranean fruit fly in California in the years 1992–94 and 1997–99. A total of 359 flies caught in monitoring traps during these years were examined at three polymorphic mtDNA restriction sites and two microsatellite loci. Composite genotypes obtained through analysis of these markers indicate at least five independent introductions of medflies into California between 1992 and 1998. Whereas the majority of specimens displayed a single mtDNA haplotype (AAA), variation of microsatellite alleles among these flies suggests at least one additional introduction in 1993 into southern California. Flies displaying the AAB haplotype sampled in 1992 both in northern and southern California shared microsatellite alleles absent in AAA flies although lacking others commonly found in AAA specimens, thus supporting the hypothesis of an independent introduction of these flies from a different source. In contrast to earlier infestations, a few specimens caught in southern California in 1993 and again in 1998 showed both mtDNA and microsatellite patterns consistent with a Hawaiian origin. Single flies collected in Santa Clara County in 1997 and in El Monte, Los Angeles County & in 1999 most likely represent a sixth and seventh distinct introduction, respectively.

The influence of genetic origin and its interaction with environmental effects on the survival of Apis mellifera L. colonies in Europe

Summary The survival and performance of 597 honey bee colonies, representing five subspecies and 16 different genotypes, were comparatively studied in 20 apiaries across Europe. Started in October 2009, 15.7% of the colonies survived without any therapeutic treatment against diseases until spring 2012. The survival duration was strongly affected by environmental factors (apiary effects) and, to a lesser degree, by the genotypes and origin of queens. Varroa was identified as a main cause of losses (38.4%), followed by queen problems (16.9%) and Nosema infection (7.3%). On average, colonies with queens from local origin survived 83 days longer compared to non-local origins (p < 0.001). This result demonstrates strong genotype by environment interactions. Consequently, the conservation of bee diversity and the support of local breeding activities must be prioritised in order to prevent colony losses, to optimize a sustainable productivity and to enable a continuous adaptation to environmental changes.

Occurrence of parasites and pathogens in honey bee colonies used in a European genotype-environment interactions experiment

Summary Diseases are known to be one of the major contributors to colony losses. Within a Europe-wide experiment on genotype—environment interactions, an initial 621 colonies were set up and maintained from 2009 to 2012. The colonies were monitored to investigate the occurrence and levels of key pathogens. These included the mite Varroa destructor (mites per 10 g bees), Nosema spp. (spore loads and species determination), and viruses (presence/absence of acute bee paralysis virus (ABPV) and deformed wing virus (DWV)). Data from 2010 to the spring of 2011 are analysed in relation to the parameters: genotype, environment, and origin (local vs. non-local) of the colonies in the experiment. The relative importance of different pathogens as indicators of colony death within the experiment is compared. In addition, pathogen occurrence rates across the geographic locations are described.

The honey bees of Ethiopia represent a new subspecies of Apis mellifera—Apis mellifera simensis n. ssp.

Honey bees endemic to the volcanic dome system of Ethiopia are described as a new subspecies, Apis mellifera simensis, on the basis of morphometrical analyses. Principal component and discriminant analyses show that the Ethiopian bees are clearly distinct and statistically separable from honey bees belonging to neighboring subspecies in eastern Africa. Considerable variation of morphological characters in relation to altitude is present in the samples under analysis, but there are no statistically separable subgroups within this population. There is no indication for the presence of more than one subspecies of honey bee in Ethiopia.

Genetic characterization of commercial honey bee (Hymenoptera: Apidae) populations in the United States by using mitochondrial and microsatellite markers

ABSTRACT Genetic diversity levels within and between the two commercial breeding areas in the United States were analyzed using the DraI restriction fragment length polymorphism of the COI-COII mitochondrial region and 10 polymorphic microsatellite loci. The western commercial breeding population (WCBP) and the southeastern commercial breeding population (SCBP) were sampled in 1993–1994 and again in 2004–2005. The goal of this study was to characterize the genetic composition of these populations and to measure potential changes in genetic diversity and composition across the sampling period. The mitochondrial DNA haplotypes C1 and C2, characteristic of the most popular bee strains (Italians and Carniolans, respectively) sold in the United States, were the dominant haplotypes at both sample dates. The frequency of Apis mellifera mellifera M haplotypes, M4, M7, and M7′, decreased during the 10-yr span. An A1 haplotype characteristic of Africanized bees was found in the SCBP from 2005. Microsatellite analysis showed there was a loss of alleles in both the WCBP and SCBP, but the losses were not significant due to simultaneous gains of new alleles into these populations between 1993 and 2005. Genetic differences that occurred between the 1993–1994 WCBP and SCBP were still detectable in these populations sampled a decade later, suggesting that these populations could be useful sources of diversity for each other in the future.

Population dynamics of European honey bee genotypes under different environmental conditions

Summary Adaptation of honey bees to their environment is expressed by the annual development pattern of the colony, the balance with food sources and the host—parasite balance, all of which interact among each other with changes in the environment. In the present study, we analyse the development patterns over a period of two years in colonies belonging to 16 different genotypes and placed in areas grouped within six environmental clusters across Europe. The colonies were maintained with no chemical treatment against varroa mites. The aim of the study was to investigate the presence of genotype—environment interactions and their effects on colony development, which we use in this study as a measure of their vitality. We found that colonies placed in Southern Europe tend to have lower adult bee populations compared to colonies placed in colder conditions, while the brood population tends to be smaller in the North, thus reflecting the shorter longevity of bees in warmer climates and the shorter brood rearing period in the North. We found that both genotype and environment significantly affect colony development, and that specific adaptations exist, especially in terms of adult bee population and overwintering ability.