EurBee board Dorothea Brückner, Germany Norberto Milani, Italy Robert Paxton, Great Britain Dalibor Titěra, Czech Republic Bernard Vaissiere, France Program consultant

The protection and use of bees´ gene pool in special programs, in rearing and in beekeeping practice cannot omit the description of the belonging to races.

The description of gene types were afore based only on pedigree records and phenotypical, morphometric analysis. The new feature for the genotype description is the microsatellite analysis.

DNA was isolated from of worker bees conserved after the taking by drying, freezing or by inserting into 75% ethanol. Isolation was carried out on ABI 3100 Nuclei Acid Preparation Station using NucPrep™ Chemistry method. The analysis was based on tests of 14 locuses assembled in four multiplexes [A29, Ap33, Ap36, Ap43],[Ap1, Ap12, Ap55],[ A37, A7, A8],[Ap113, Ap16, Ap19, Ap34]. In this way > 600 bees were analysed various geographical races [carnica, mellifera, ligustica] from various regions.

The applied method offers data for very detailed discrimination of distances among populations.

The results were compared and completed by morphometric analysis of wings using DAWINO method. The method of microsatellite analyzes is a good tool for very detailed discrimination of distances among populations.

Supported by grant project NAZV 1G46032.

Non-Apis bees

Agroecology, Department of Crop Science, University of Göttingen, Germany: isteffa©gwdgd.de

Human destruction and fragmentation of natural or seminatural habitats and the creation of intensively managed agricultural habitats are major causes for the loss of biodiversity in terrestrial ecosystems. However, it is not well understood to which extent and at which spatial scales pollinator species respond to the resulting loss of habitats and changes in landscape structure. Further it is unclear how biodiversity change affects ecosystem functioning and services. Crop pollination is an ecosystem service of significant economic value, and there is increasing evidence that wild pollinators contribute to production in several crops. In this context I will present key results and ongoing interdisciplinary research projects.

Bumblebee behavioural and sensory diversity under threat by indiscriminate trade?

Lars Chittka, Thomas C. Ings, Nigel Raine, Peter Skorupski

School of Biological & Chemical Sciences, Queen Mary, University of London, Mile End Road, London E1 4NS, E-mail: l.chittka©qmul.ac.uk

The European bumblebee species Bombus terrestris has recently become a model to study the evolution of sensory traits and behaviour, because of the intriguing variation that occurs between populations of this species. Island and mainland populations from Corsica, Sardinia, Britain, the Canary Islands, Germany, France, Turkey and Israel differ in spectral tuning of photoreceptors, floral colour preferences, learning behaviour, foraging performance, phenology, and colour coat. As we are beginning to understand the ecological and evolutionary reasons for this diversity, these between-population differences are already under threat because commercial breeders ship colonies from these populations in large numbers to destinations where they are not native. This is because bees of this species are popular with growers of commercial greenhouse crops, such as tomatoes. However, non-native bumblebees might easily establish at non-native localities, interbreeding with native populations. We present a risk assessment of such commercial displacements, and viable strategies to minimise the dangers of population homogenisation.


Communication in eusocial bees: comparison of collective foraging

Koos Biesmeijer

University of Leeds, j.c.biesmeijer©leeds.ac.uk

Social bee foragers can collect food either based on their own decisions, based on information obtained from nest mates or a combination of both. Honeybees, stingless bees, and bumblebees differ strongly in the behavioural mechanisms they have developed to explore for and to exploit food sources and in this talk I would like to try to compare the different groups of social bees using a single coherent concept (see Biesmeijer and de Vries 2001). I will characterize individual foragers as scout, recruit, inspector, reactivated forager, employed forager, unemployed experienced forager, and novice forager. I will emphasise on stingless bees and bumblebees, but compare them to honeybees throughout.


Mating in bees: unique behaviour by which males apply pheromones onto female antennae

Dieter Wittmann, Matthias Schindler, Betina Blochtein*, Dirk Barouz

INRES, Department of Landscape Ecology, University Bonn, Melbweg 42 53127 Bonn, Germany; * PUC-RS, Porto Alegre Av. Ipiranga 66

wittmann©uni-bonn.de

In several taxa of bees males have independently evolved modes of mating behaviour through which sex-pheromones are applied directly onto the antennae of females.

In Anthophora plumipes the males’ middle legs are elongate and bear long brush-like hairs, but no odour glands. Before mounting the female the male brushes secretions from glands in the abdomen onto his hind legs. They then transfer the odour to the middle legs and finally they brush this secretion during mating onto the female’s antennae.

In megachilid and Coelioxys bees of the New and Old World males have modified front legs which bear the odour glands in the tibia and/or in the basitarsus. To apply the contact sex-pheromone, a megachilid male mounts the female, holds the female’s antennae with special structures of his mandibles, covers her compound eyes with blind shields present on his front legs and then secretes the paste like pheromone from pores in his front legs onto the antennae.

In some genera of New and Old World Xylocopa, males have similar structures on their front legs and exhibit similar behaviour as described for megachilid bees. However, in some species of Xylocopa the middle legs, which are modified to hold the female’s antennae and secrete the odour, are of importance in this mating behaviour.

In several species of the cuckoo bees Nomada, males have pores of pheromone glands in various segments of the flagella. During mating the male winds it’s flagella around the female’s antennae and slowly pulls them upwards to apply the paste like secretions onto the female’s antennae. The significance of this chemical communication is discussed.

Institute of Landscape Ecology, E-mail: kuhlmmi©uni-muenster.de

The palaearctic Colletes succinctus species group comprises twelve species with three of them occuring in western Europe: C. succinctus (Linnaeus) (oligolege of Calluna and Erica), C. hederae Schmidt & Westrich (oligolege of Hedera) and C. halophilus Verhoeff (oligolege of Asteraceae, especially Aster tripolium). Their close morphological resemblance, the minute genetic differences and distribution patterns suggest that these three species are closely related and probably very recently diverged taxa.

For C. halophilus, an endemic species of the coast of the southern North Sea and the English Channel, a hypothetical scenario of speciation was developed. It is based on the recent postglacial landscape change in the southwestern Netherlands that is assumed to be the centre of origin of this species. The effect of landscape change on the availability of flowers as pollen source and the influence of parasitic cuckoo bees is assumed to be the driving force for the speciation of C. halophilus.

Osmia lignaria is a spring-flying solitary bee that very effectively pollinates fruit tree flowers. Methods have been developed to rear and manage populations of this bee for orchard pollination. The establishment of this or any other agricultural pollinator roughly follows a series of steps. First, information on the basic biology of the species is needed, including its life cycle and developmental biology, nesting behavior, population dynamics and pollinating efficacy. At the same time, the basic biology of the pollinator’s parasites and predators should also be studied, as well as the reproductive biology of the target crop. The information acquired through these studies is combined to design a management system. This system should provide guidelines on rearing methods, nesting materials, population densities required for adequate pollination and control against parasites and predators. The management system should then be tested in field trials, and modified accordingly. Finally, the information should be delivered to the agricultural industry. Only pollinators for which a reliable supply of bees can be secured are likely to become established as crop pollinators. In this presentation, we provide an overview of the process that has led to the establishment of O. lignaria as an orchard pollinator.

Universidad Autonoma de Yucatan, E-mail: qeuan©tunku.uady.mx

Stingless bees are amongst the most important pollinators in tropical and subtropical environments; however, little attention has been given to their potential use as pollinators in confined environments. We have studied the adaptability and pollination efficiency of these species: Trigona nigra, Scaptotrigona pectoralis, Melipona beecheii and Nannotrigona perilampoides.

Here we report results on Nannotrigona perilampoides. We first evaluated its acclimation, foraging behavior, and pollination efficiency using tomato (Lycopersicon esculentum) and habanero pepper (Capsicum chinense) in greenhouses. The pollination efficiency against mechanical vibration (MV) and no pollinator (NP) was compared through the percentage of fruit set, weight of individual fruit, kilograms of fruit produced per square meter, and the number of seed per fruit. In this first study we collected evidence that N. perilampoides performed as well as MV and better than NP.

In a second experiment, we evaluated the efficiency of N. perilampoides on large-scale greenhouse pollination against bumble bees. In Yucatan, the bumble bee Bombus impatiens has been the species mainly used for pollination in greenhouses. In this experiment we also used C. chinense. The results showed that the plants pollinated by N. perilampoides set more fruit and the production in kg per m2 was better than those pollinated by B. impatiens mainly as a result of the capacity of stingless bees to continue foraging at high temperature and humidity and because bumble bees caused damage to the flowers.

These results suggest that for neotropical enclosures stingless bees can be a good pollinator alternative compared to temperate bumblebees and hand pollination. More studies need to be conducted to increase the production of colonies for pollination purposes.

Department of Agrienvironmental Sciences and Technologies

University of Bologna - Viale G. Fanin 42, 40127- Bologna, Italy. E-mail: bmaccagnani©entom.agrsci.unibo.it

The role of ecological infrastructures in pear orchards as food sources functional to pollinators management has been investigated. The researches focused, in particular, on the possible function of early flowering boundaries and edgerows in order to anticipate the release of the pear pollinator Osmia cornuta (Latreille) prior to pear (Pyrus communis Linnaeaus cv. ‘Abbé Fetèl’) blooming. Trials were run in the years 2004-2005 in two orchards, one with a Prunus spinosa Linnaeus edge, the other with strips of Brassica oleracea Linnaeus and Brassica napus Linnaeus, sowed during the previous autumn. Four nesting shelters were placed in each orchard, and 200 females and 400 males were released per shelter. Ten females per shelter, and their nests, were identified. The functionality of the two kinds of early flowering food sources in allowing nesting activity’s initiation was compared in terms of number of larval cells produced in the first part (till 50%) and in the second part (till 100%) of the flowering. The number of cells produced in both pear orchards was also compared with data collected in the years 1998-2000, when O. cornuta cocoons had been released at the beginning of pear flowering. Different safe measures were adopted and compared to preserve females from pesticide sprays at the end of pear flowering. Results are discussed.

Department of Evolutionary Biology, University of Vienna, Austria, E-mail: tkis21©gmx.de

Size variation amongst eusocial bees is most profound in the genus Bombus. It has a strong impact on individual behaviour and colony organization. The goal of this study was to elucidate whether size variation has an impact on morphology and light sensitivity of the visual system of Bombus terrestris. After a comprehensive analysis of the compound eyes and ocella we discovered distinct morphological differences in the eye structure, with larger individuals showing an increase in size and number of ommatidia as well as an increase in ocella diameter. In a behavioural experiment we found that differences in body size have a strong impact upon the light sensitivity of the visual system, with larger bees being able to fly at significantly lower light intensities. These data suggest that larger individuals are able to forage earlier at dawn and later at dusk compared to their smaller nestmates, thus being yet another possible factor contributing to the size related division of labour in bumblebees.


Rearing Bombus lapidarius L. (Hymenoptera) in laboratory

Vladimir Ptacek

Masaryk University, Faculty of Science, IEB, Kotlarska 2, 637 11 Brno, Czech Republic,

When a method of laboratory rearing of Bombus terrestris L. was applied as the standard in B. lapidarius, the rearing conditions were fully acceptable for B. lapidarius queens. Ways of starting colonies, their development, numbers of individuals, were similar in both species. The outstanding feature of most lapidarius queens was their willingness to accept the cocoon(s) of B. terrestris and even the very young terrestris worker as a helper in the care of brood at the very beginning of colonies. On the contrary from B. terrestris, lapidarius colonies did not produce young queens after they were made orphan, what in terrestris is the rule.

Lapidarius colonies did produce young queens regularly. Those freshly emerged, fed pollen first to develop their fat body, and from the 6th day onward they were ready to mate. They did mate in comparative small copulation rooms (40 litter aquarium covered with mesh) at the window (light). Males produced in laboratory copulated regularly, whereas those from outside did not.

Unlike in B. terrrestris, where currently 70-80 % of queens survived the period of 3-6 months of cold storage, in B. lapidarius 75–87 % of queens died under the same conditions. Moreover those, which survived were hardly able to start colonies. The process of entering the diapauses remains to be solved.

This presentation was possible thanks to the financial support from NAZV CR No: IR440114.


Effects of different starting methods on Colony foundation of Bombus terrestris L. queens

Fehmi Gurel, Ayhan Gosterit

Akdeniz University, Faculty of Agriculture, Antalya, Turkey

E-mail: fgurel©akdeniz.edu.tr

In extensive commercial rearing, one of the main success criteria is colony foundation ratio of hibernated Bombus terrestris L. queens. The effects of different starting methods on the colony foundation of hibernated B. terrestris queens were investigated. Total of 350 hibernated queens obtained from commercial company were used. Each queen was treated with CO₂ and placed separetely in the starting box. Queens were submitted to four starting methods: (qtw) queen with one B. terrestris worker, (qhw) queen with one honey bee worker, (qtp) queen with one B. terrestris pupa and (q) only queen. Queens were fed ad libitum with a sugar syrup and fresh frozen pollen and kept in climate rooms at 27 ^oC ± 1 and 65 ± 10% R.H. The highest egg laying ratio was found in the qtw group (82.8%) followed by qtp (72.8%) , qhw (57.1%) and q (35.7%). In qtw, qtp, qhw and q groups, queens started laying eggs 11.4 ± 0.8, 11.7 ± 1.3, 15.4 ± 0.9 and 20.7 ± 1.7 days respectively after they were placed in the starting boxes. Number of egg cells in the first brood was determined to be an average of 4.4 ± 0.2, 3.5 ± 0.2, 3.3 ± 0.2 and 2.7 ± 0.3 in qtw, qhw, qtp and q groups, respectively. Results showed that adding a B. terrestris worker is the best starting method in bumblebee rearing.

The effects of weight of queens after diapause on colony development in Bombus terrestris were investigated in this study. The weight of 57 queens was determined to be an average of 0.792± 0.012 g after diapause. Approximately 82% of the queens laid eggs and 64% of egg laying queens produced worker. No significant statistical correlation was found between the weight of the queen after diapause and colony development patterns in Bombus terrestris. Only the correlation between the weight of the queen after diapause and the number of workers produced in the first brood was found to be significant (p <0.05, r = 0.42, n = 30). Many factors affect on the variation in colony development patterns and the colony foundation ratio of queens during the year round rearing of Bombus terrestris. However, results showed that the weight of the queens after diapause did not affect colony development in Bombus terrestris.

About 300 bumblebee (Apidae: Bombus) species in 38 subgenera are known in the world. Previous morphological studies showed that 31 bumblebee species in nine out of 16 European subgenera can be found in Slovenia. The aim of our study was to obtained characteristic of wing venation, for assessment of morphological structure and diversity of bumblebees in Slovenia. Over 350 bumblebees from 84 different localities in Slovenia were collected. Right-sided forewings were removed, scanned and analysed. The co-ordinates of 19 vein crosses were measured and used in the calculation of 35 characteristics. Species means and standard deviations of the morphometric characters were analysed. Statistical analysis using multivariate discriminant analysis and a cluster analysis of the Mahalanobis distances between centroids showed significant (P<0,05) differences between species. With discriminant analysis we classified into right species more than 96% of bumblebees. The most informative characters was dumb-bell index (R2 0,70). Wing venation characters could be also useful tool for species determination, especially for species which are hard to identify.

Agroscope FAL Reckenholz, Swiss Federal Research Station for Agroecology and Agriculture, Reckenholzstr. 191, 8046 Zürich, Switzerland, E-mail: roger.konrad©students.unibe.ch

Potential effects of genetically modified (GM), insect-resistant crops on non-target organisms are widely discussed. Honey bees generally have to be tested for effects of such plants or their insecticidal products and few data are also available for bumble bees. However, solitary bees have not been tested to date, although they form a very diverse group of pollinators and are both of ecological and economical importance. Furthermore, they differ from honey bees in various aspects of their biology (e.g. provisioning of developing larvae).

We conducted feeding assays under defined indoor conditions with larvae of the red mason bee Osmia rufa. Larvae were reared either on pollen provisions that were bee-collected on a transgenic insect-resistant oilseed rape (producing the protease inhibitor OC-I) or on bee-collected pollen from control plants where a transgene compound (i.e. the Bt-toxin Cry1Ab, the lectin GNA and OC-1) had been manually applied in purified form. During larval development and the subsequent hibernation period, life history and viability parameters were recorded. The data from two consecutive experimental periods (summer 2005 and 2006) will be combined. Data from 2005 revealed that 0.1% GNA in the larval diet resulted in a significantly lower food conversion (i.e. conversion of provision weight into larval weight). No significant departure from the control was observed for all other treatments and parameters tested. Our experimental design proved to be suitable for testing direct effects of transgenic plants or their purified insecticidal products on solitary bees and could also be adopted for testing agrochemicals.

Bombus terrestris is a bumblebee species whose males exhibit patrolling behavior during their mating phase of life. While patrolling, males laid scent marks to attract conspecific females. The scent is produced by the cephalic part of the male labial gland. Little is known about changes in the scent production during the male life span.

Labial glands of males of different age were dissected, extracted with hexane, analyzed and the components were quantified. Chemical analyses revealed significant age-dependent changes. Very young males (0-1 day) posses only traces of compounds in the gland extracts (1-20 µg/gland). During 2-7 consecutive days, amounts of compounds increase rapidly (up to 7 mg/gland) and decrease slowly during the following 30 days (down to 7 µg/gland). Terpenic alcohols (2,3-dihydrofarnesol and geranylcitronellol) appear first in the extract followed by aliphatic compounds (ethyl dodecanoate and hexadecanol). Glands of old males (33 days) contained hydrocarbons and esters of terpenic alcohols such as 2,3-dihydrofarnesyl dodecanoate.

EAG and GC-EAD experiments were performed on antennae of virgin queens. The EAG recording showed maximum responses to extracts of glands of 2-7 days old males. GC-EAD analysis revealed six EAD active components of the labial gland extract. Males’ antennae responded to the same components as those of queens.

Data show that pheromone biosynthesis starts immediately after eclosion and reaches its maximum about 4th posteclosion day. It was also confirmed by the transmission electron microscopy of the cephalic part of the labial gland.


Multitemporal investigation of Giant honey bees (Apis dorsata) migration in Chitwan area (Nepal) by means of remote sensing

M. Hirschmugl¹, E. Huettinger², G. Kastberger³, W. Sulzer<sup>1

1</sup> Institute for Geography and Regional Science, University Graz/Austria

² BFL-Institute of Apiculture, Lunz/Austria

³ Institute of Zoology, University Graz/Austria
E-mail: wolfgang.sulzer©uni-graz.at

The study area is located in the district of Chitwan (Nepal). The Giant Honey Bee swarms (Apis dorsata) migrate in autumn and winter to the southern sub-tropical part of Nepal. The main focus of the inter-disciplinarily project is to analyze the occurrence and distribution of bees (done by field work) together with the spatio-temporal (December, February and April) expansions of the crop fields (by LANDSAT-ETM+), to get out the relationship of bee migration and special food offer. The traditional way of monitoring by doing field work in a large area with parts far off roads and not easily accessible within the monsoon season is very difficult and expensive, sometimes not even possible. Therefore, a good alternative to get data about these areas is by means of remote sensing.

Due to lack of a public geodata base, the whole range of data acquisition (digital maps, Remote sensing data), pre-processing (building up a GIS data base (digital elevation model, digital maps, land use and land system maps) and analyses within Erdas/Imagine must be applied. The workflow and its results will be documented in the presentation paper.
Communication precision of foragers in the stingless bee Scaptotrigona mexicana

D. Sánchez, F. B. Kraus, M. de Jesús Hernández, R. Vandame

ECOSUR, E-mail: bkraus©tap-ecosur.edu.mx

For eusocial bees the precision of communication of food sources, i.e. the proportion of recruits that reach a communicated food source, is a crucial parameter for colony fitness. However, only little is known about the precision of stingless bee communication systems. Here we examined the impact of experience of workers and distance of the food source on the precision of the food communication system of Scaptotrigona mexicana.
The experiments were conducted by training bees to a three dimensional artificial patch at varying distances from the colony. We recorded the choices of individual recruited foragers, either being newcomers (newly arriving recruits) or reactivated (foragers that had previously visited the feeder). We found that the average precision of newcomers (95.6%±2.61%) was significantly higher (p<0.001) than that of reactivated bees (80.2%±1.12%). This effect, in which the newly arriving bees were significantly more precise, was found at all four experimental distances tested (p=0.004). While this might seem counterintuitive on the first sight, this \"loss\" of precision can be explained by the tendency of experienced workers to explore nearby areas to find new rewarding food sources, after they initially had learned the exact location of the food source. Increasing the distance from the colony had no significant effect on the precision of the foraging bees, neither for newcomers nor for the reactivated ones. Thus our data show that experience, but not the distance of the food source, affected the patch precision of S. mexicana foragers.


How bees get rid of ants?

Orawan Duangphakdee¹, Nikolaus Koeniger² , Siriwat Wongsiri¹ and Sureerat Deowanish <sup>1

1</sup> Center of Excellence in Entomology: Bee Biology, Biodiversity of Insects and Mites, Department of Biology, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand, ² Institut für Bienenkunde (Polytechnische Gesellschaft), Fachbereich Biologie und Informatik der J.W.Goethe-Universitat Frankfurt am Main, Karl-von-Frisch-Weg 2, 61440 Oberursel, Germany

The bees collect the resinous materials from various plants for many purposes. Apis mellifera worker bees collect plant resin for plugging cracks and holes in their nest cavity. Apis florea and Apis andreniformis apply the plant resin around the nesting branches. In contrast to Apis species, stingless bees are using wax mixed with abundant propolis to build an envelop (involucrum) surrounding the nest. We designed a bioassay to test the response of the predatory ant species, weaver ants, Oecophylla smaragdina, and red wood ants, Formica polyctena, toward bee materials from Genus Apis and Trigona. The bee materials were found to be highly repellent against weaver ants in the raw materials. Interestingly, pentane extracts exhibited repellent activity toward weaver ants whereas the acetone and methanol extracts were no repellent activity. On the contrary, the bee materials resulted in very low repellent activity against red wood ants. Perspectives and directions for further research on the active components are discussed.

Vespa orientalis is a threat to beekeeping in the countries of the Near East. Apis mellifera syriaca, the indigenous honey bee of Syria, Lebanon, Palestine, Israel and Jordan, is a bee adapted to dry-hot regions and reported to have superior ability to withstand attacks by Vespa orientalis. As a particular reaction, bees are said to commence flight during wasp attacks. We investigated, whether bees would decrease flight activity in response to presenting tethered V. orientalis wasps at a distance of 5 to 10 cm to the colony entrance. In two years we investigated a total of 29 A. m. syriaca colonies, 16 kept in traditional and 13 kept in modern hives, and of 13 colonies of A. m. ligustica in modern hives. Outflying bees were counted over 5 min, then a tethered wasp was slowly waved before the entrance for 5 min, and outflying bees were again counted over the following 5 min. The experiments confirmed previous observations of a reduced flight activity of honey bees elicited by the attack of predatory wasps. The reduction was present in both bee subspecies tested, but it was more pronounced in A. m. syriaca (48%) compared to an imported strain of A. m. ligustica (22%) indicating a better suitability of the indigenous bees of Jordan, where attacks by V. orientalis are a common threat during summer and autumn. The results further indicate, that traditional bee hives (A. m. syriaca, 48%) might be better suited to withstand attacks compared to modern Langstroth hives

Osmia lignaria is a univoltine, spring-flying solitary bee from North America. Development from egg to adult occurs during spring and summer. Eclosed adults remain in their cocoons through the winter in a diapausing stage, and emerge the next spring as temperatures increase. Chilling is required to complete diapause development. We define pre-wintering as the period during which cocooned adults are still exposed to warm temperatures in late summer and early fall. Because the timing of the onset of cold temperatures in autumn varies widely among years, pre-wintering duration may be highly variable from year to year. Our objective in this study was to establish whether pre-wintering duration affects diapause development and fitness in O. lignaria. We exposed females to three different pre-wintering durations (5, 30, 60 days) to simulate years with early, mid and late winter, and measured respiration rates, ovary maturation, weight loss, fat body depletion and lipid levels throughout pre-wintering and wintering. We also measured survival and vigour at emergence. Pre-wintering duration affected diapause development, and we obtained significant differences among treatments in all variables measured. Females exposed to long pre-wintering periods lost more weight and used up their fat body reserves. These females were less likely to survive the winter or were less vigorous in spring. We discuss the relevance of these results for fitness, nesting activity and the dynamics of O. lignaria populations.

.

Environmental Hazards to Honey Bees

LAVES Institut für Bienenkunde Celle, E-mail: martina.janke©laves.niedersachsen.de

In recent years colony losses reiterated in Germany. Besides Varroosis many other causes like other bee diseases, nutrition supply as well as effects of pesticides have been discussed.

A chronic feeding study was conducted to find any indications to what extent negative effects of pesticides in sub-lethal doses can be defined from effects of other stressors (pathogens, drugs, mix of plant protection products, malnutrition of proteins) or if there will be found any interactions or coactions.

In screening programme effects of chronic dietary exposure to sub-lethal doses of the insecticide imidacloprid were studied in honeybees being loaded with a potential stressor (Varroa destructor, Nosema apis, drugs, lack of pollen supply). The results confirm a chronic oral toxicity of imidacloprid at concentrations which have in several previous studies reported to be toxic to bees (100 ppb). However, no indications were found for significant differences in sensitivity to imidacloprid between bees being loaded with different stressors and control bees.

Results confirm previous findings that optimizing of protein supply can soften negative effects of stressors. In addition it became apparent that bees from different colonies of the same apiary which were fed in parallel varied in sensitivity.

Semi-field experiment was conducted to asses the risks of mixing plant protection products by simulating commercial application during blooming on bee colonies foraging in commercial seed dressed rape with potential residues in nectar and pollen.

No adverse effects on mortality or on development of exposed bee colonies had been found when bees foraged on rape of dressed seeds and plants were sprayed with one single plant protection product alone (pyrethroid resp. azol-fungicide) or in combination (tank mix pyrethroid plus azol-fungicide).

From the findings of chronic feeding tests and semi-field test it can be concluded that imidacloprid used as standard seed dressing formulation will pose no risks to honeybees.

Pesticide transport with nectar and pollen loads following treatments in different blooming cultivations

J.Pistorius, K. Wallner see page 146 for abstract

Universität Hohenheim, Landesanstalt für Bienenkunde D-70593 Stuttgart/ GAB Biotechnologie GmbH, 75223 Niefern-Öschelbronn, E-mail: jenspistorius©gmx.de

AFSSA, E-mail: mp.chauzat©afssa.fr

Twenty five apiaries, evenly distributed in continental France have been followed for 3 years in order to monitor weakness of honey bee colonies (Apis mellifera L.). Five colonies randomly selected in each apiary (total: 125 colonies) were visited four times per year during the whole survey. Depending on the season, the following samples were collected at each visit: pollen from traps, adult bees, foundation wax or honey. This three year study has been completed in 2005. Multiresidue analyses were performed on all of the apicultural matrices in order to detect residues of various pesticides including forbidden substances. Pesticide residues were found in all of apicultural matrices. For example coumaphos residues were found in pollen, carbofuran in honey and fungicides in honey bees.

In pollen, honey and honey bees specific analyses were conducted to search fipronil, imidacloprid and some of their metabolites residues. Fipronil and/or metabolite contents were superior to the limit of detection in 16 samples out of the 81 collected in 2003. Residues of imidacloprid and/or 6-chloronicotinic acid were found in 69% of pollen samples collected in 2003. Imidacloprid contents were quantified in 11 samples out of 81 with values ranging from 1.1 to 5.7 µg/kg.

The different steps of the French regulatory scheme for plant protection including post-release field controls are detailed. The occurrence of the residues found in the present study is commented in regard to these legal requirements.


An investigation of potential long-term impact of clothianidin seed-treated canola on honey bees, Apis mellifera L.

C.D. Scott-Dupree and G.C.Cutler

Department of Environmental Biology, Ontario Agricultural College, University of Guelph, Guelph, Ontario, Canada, N1G 2W1, E-mail: cscottdu©uoguelph.ca

The past decade has witnessed considerable debate over the impacts of chloronicotinyl insecticides on honey bees, Apis mellifera L. Few studies have investigated potential long-term effects after chloronicotinyl exposure. We conducted a 130-day investigation to ascertain effects on honey bee colonies after exposure to canola grown from seed treated with the chloronicotinyl clothianidin. Colonies were placed in the middle of 1-ha untreated or clothianidin seed-treated canola fields for three weeks during bloom, and thereafter moved to a fall apiary. There were 4 treated and 4 untreated fields, and 4 colonies per field, giving 32 colonies total. Bee mortality, worker longevity, and brood development were periodically assessed in each colony throughout the study. Samples of honey, beeswax, pollen and nectar were regularly collected from colonies and analyzed for clothianidin residues using HPLC with MS/MS detection. Although sporadic treatment or site effects were found on various dates, essentially no differences in bee mortality, worker longevity, or brood development occurred throughout the study. Weight gains of and honey yields from colonies in clothianidin-treated fields were not significantly different from those in control fields. While clothianidin residues were detected in honey, nectar, and pollen from colonies in clothianidin treated fields, maximum clothianidin concentrations detected were 8- to 22-fold below the NOAEC. The results suggest that honey bee colonies will be unaffected by exposure to clothianidin seed-treated canola during bloom.


Sublethal effects of fipronil on the ability of foragers to orientate in a complex maze

Axel Decourtye

ACTA, E-mail: axel.decourtye©acta.asso.fr

The insecticide fipronil was recently suspected to disorientate honeybees during their returning flight to the hive. In response to whether fipronil may disorientate foragers, its impact on orientation in a maze under outdoor conditions was examined. The maze consisted of a 4 × 5 matrix of identical cubic boxes, with each side carrying a hole in its centre where bees crossed (Zhang et al., 1996). Bees had to fly through a sequence of nine boxes to reach the goal – a feeder containing a reward of sugar solution. During training, the foragers were taught to associate a green mark to the reward that was moved step by step along the path. After foragers were trained and individually marked, their ability to find their way according to the presence or the absence of green marks was measured. Along the path, three boxes constituted a decision point: the bee had to choose between a marked hole (correct path) and unmarked hole leading to dead end. Correct and wrong decisions, turn back, and course duration were recorded. Training and test phases were carried out following two feeding conditions: syrup delivered in a feeder placed outside the maze (before and after treatment); syrup added with 1 µg kg-1 (quantified by HPLC-MS) fipronil (treatment). Data of each period were obtained from different bees.

The rate of foragers entering the maze, and so responding to the mark placed at the entrance, was largely reduced in fipronil-fed animals. Before and after treatment, 86-89% of bees equally flied through the whole path and arrived to the goal without mistakes (P=0.35). The rate of fipronil-treated bees making path without mistakes decreased to 60%. The performance was lower than controls (P<0.01). In parallel, the rate of bees with unsuccessful searches for goal notably increased with treatment (34% in treated bees versus 4% in control bees). So, the orientation capacities of foragers in a complex maze were highly affected by fipronil.

CSL, York, UK, E-mail: h.thompson©csl.gov.uk

The systemic properties of some pesticides used in agriculture and horticulture (e.g. granules, seed treatments, soil drenches) means that the laboratory-based acute toxicity studies undertaken for honeybee risk assessment may be less relevant for assessing the potential for adverse effects in the field. Pesticides with systemic properties may be expressed in pollen or nectar and are thus available to foraging bees at low levels but for longer periods of time depending on the flowering of the crop. Foraging bees returning to the hive may also bring contaminated nectar and pollen used to feed brood within the colony. Current OECD test guidelines only allow for short term exposure of adult worker bees (a single contact dose or a 4 hr oral exposure) with assessments of mortality for up to 48 hrs. The relationship between the toxicity of such acute exposures and chronic toxicity in honeybees is not well established. Chronic (10 day) exposure studies have been undertaken for a limited range of chemicals but with widely differing results. This paper will consider the design of a suitable 10-day LC50 test procedure and assessment of the toxicity of a range of chemicals to worker bees. It also will consider data generated toxicity of pesticides to honeybee larvae to determine whether they are protected by current adult honeybee studies.


A NEW LARVAL IN VITRO REARING METHOD TO TEST EFFECTS OF PESTICIDES ON HONEY BEE BROOD

P. Aupinel, D. Fortini, B. Michaud, J.N. Tasei, J.F. Odoux

A new in vitro rearing method of honey bee larvae was devised to assess the effect on brood of any substance that can reach the hive with nectar and pollen. This method could be used in the risk assessment process since it meets with the following characteristics:

low mortality rate when no treatment is applied,

standardisation of the test,

easiness of carrying out,

sensitiveness to treatment,

precise control of the ingested doses of diet and pesticide,

control of larval mortality for each larval instar,

control of pre pupae weight,

measurement of adult emergence rate.

The method enables the study of 1/ lethal effects (calculation of LD50) 2/ sublethal effects (prepupal weight, duration of development, adult morphology and behaviour).The method can be used either to study acute effects by applying contaminated diet to one particular instar, or to investigate chronic effects by providing each day the larvae with the test substance.

The use of our rearing method is illustrated with two insecticides:

Dimethoate considered as a reference insecticide in toxicological tests on adults,

Diflubenzuron which belongs to the IGR group.

ACTA, E-mail: axel.decourtye©acta.asso.fr

We have studied the effects of insecticide fipronil in the honeybee (Apis mellifera L.) in semi-field conditions. Each modality was conducted twice, with a colony of about 20 000 workers and a fertile 1-year-old queen, set in a tunnel. The food was delivered with two artificial feeders filled with sucrose solution and multi-floral pollen contaminated (1 ppm, 100 or 10 ppb) or not by fipronil. Any dead bees found on the ground were counted daily. All anomalies in development and behavior of the honeybee colonies were recorded. The colonies were visited to assess brood surface and food quantities (honey and pollen). Larval development was measured using the method of Oomen et al. (1992). No impact of fipronil at 10 ppb was found on the survey, the foraging activity of workers adults and on larval stages, whilst impact was found with others concentrations. Lethal effect on workers bees were observed with two higher concentrations. The contamination of pollen with fipronil at 1 ppm induced a drastic decrease in the foraging activity on the food source. Negative effects of fipronil were also noted on larval stages.


Effect of Bt-corn pollen on the development of honeybee larvae and resistance of adult workers

L. Békési¹,B.Darvas², A.Székács², É. Lauber²,E. Zajácz¹, E.Szalai Mátray<sup>1

1</sup> Research Institute for Animal Breeding and Nutrition, Gödöllő, Hungary. E.mail: bekesi©katki.hu ²Plant Protection Institute, Budapest, Hungary

For the time being, plantation of GM crop has been suspended in Hungary. Supposed coexistancy of transgenic and isogenic plants is a new challange for bee breeding and honey production.

According to recent reports, CryAb toxin expressed in pollen by MON 810 corn, does not cause acute toxicity and not reduce development of hypopharingeal gland of workerbees either (Ramirez-Romero et al., 2005). However no data are available about the effect of Bt-pollen to larval development in vitro and resistance of honeybees.

In our experiments, larval food containing 10 % of Bt corn pollen, 10 % of fresh bee bread was offered in royal jelly to 4 day old larvae maintained in U-shape microplates in vitro. The plates were incubated at 35 ˚C with 90 % relative humidity. The same feed, but with isogenic corn pollen was given to controls. Controls fed with fresh bee bread containing feed was also applied. On the 8th day, before pupation, the larvae were removed, weighed, and deepfreezed for toxin analysis.

Adult worker bees were placed into wire cages by 20 in each and maintained in incubator at 25 ˚C and 90 % relative humidity. As feed they were given honey twice a day, that contained 10 % Bt-corn pollen and 10 % fresh bee bread. The feed of the controls contained isogenic corn pollen. During 4 week period, mortality rates were registered and Nosema spore analysis of dead bees was performed.

According to our preliminary results, Bt-corn pollen containing feed seems to slightly reduce in vitro larval development. Further more we noticed reduced survival rate and increased Nosema spore count in caged, Bt-pollen fed adult bees.

AFSSA, E-mail: m.aubert©afssa.fr

In 1997, several beekeepers reported that colony death and weakness were the consequence of the use of imidacloprid for seed dressing. This product is indeed very toxic for bees as confirmed by laboratory and semi-field tests. However, several field studies were conducted in France and elsewhere with colonies kept close to cultures issued from seeds dressed with imidacloprid. None has shown any adverse effect of this product. This has been confirmed by some bee-keepers that testified that they suffered no adverse effect from such cultures. Additionally, observations in France revealed imidacloprid in pollen loads of several surveyed apiaries with no acute adverse effects on them.

An expert panel gathered by the French Ministry of agriculture studied several scenarios of contamination of bee colonies by imidacloprid. Experts concluded that several PEC/PNEC ratios were very preoccupying. However the experiment we had conducted on colonies for mimicking the natural consumption of contaminated nectar (the most risky scenario according to the expert panel) did not show any adverse effect on the tested colonies. This apparent discrepancy probably originates from the security margins used in the above mentioned risk analysis.

We conclude that, if contamination by imidacloprid from sunflower cultures issued from treated seeds may have occurred simultaneously with a period of colony losses as described by several French bee-keepers, such occurrence has not been observed systematically, and no negative impact on bee colonies of the use of Gaucho® has been experimentally demonstrated in the field.


The pragmatism of honey bees as environmental bioindicators

Claudio Porrini¹, Anna Gloria Sabatini², Piotr Medrzycki², Fabio Sgolastra¹, Laura Bortolotti²

1DiSTA – area di Entomologia, University of Bologna, Italy (cporrini©ento.agrsci.unibo.it)

2C.R.A. Istituto Nazionale di Apicoltura, Bologna, Italy

The goal of environmental pollution control is the protection of human health. Most of applied monitoring methodologies are based on chemical-physical procedures that provide, through their high analytic capacity, exact data regarding the concentration of contaminants in the environment. However, this information is not complete because only a part of the pollution has a toxicological importance for the organisms. There is no data about assimilable quantity or biological effects of the pollutants. For this purpose the biological indicators (e.g. honey bees) can be very useful. They can respond, in accordance to the nature of the contaminant, with population variations, behaviour changes or they can bio-accumulate it, by intercepting and assimilating the bio-available fraction. Anyway these assumptions are not considered by the environmental control agencies probably because the evaluation of bioindicator goodness (particularly honey bees), is based only on the comparison with chemical-physical instruments or with environmental markers. This way, for example the fact that honey bees are present and visit all the environmental sectors, is not taken into account. Different methods and instruments, indeed, in relation to their prerogatives, find only a part of the environmental pollution that not necessarily must coincide. The chemical-physical system can be defined as eidetic, because it finds the total amount of the contaminants, while the honey bees find the contaminants in pragmatic way, i.e. considering the bio-availability fraction and the related effects. Integrated monitoring is required to obtain more complete information about environmental pollution.

INSTITUTE FOR DIAGNOSIS AND ANIMAL HEALTH, BUCHAREST, ROMANIA, E-mail: bianu.elisabeta©idah.ro

Substances hazardous to bee (e.g. heavy metals, pesticides, agrochemicals etc.) induce changes in bee activities or colony performance. Industrial exhaust gases contain a variety of mineral acids and heavy metals. The aim of this study was to investigate the cadmium, lead and zinc levels in sample of honeybees, mellifera flora and honey in a polluted area from Romania compared with the ones in the clean area. The heavy metals pollution was produced by a non-ferrous metal processing plant. 30 samples of honeybees, 25 samples of mellifera flora and 20 samples of honey were analysed. The samples were harvested from hives situated at 10-15 km away from the plant.

The concentration of cadmium, lead and zinc were measured by atomic absorbtion spectroscopy after mineralization of the samples in a microwave digestion system.

The high amounts of heavy metals were found in honeybees comparing to the control area. The mean level of Cd was 6,2 mg/kg, Pb level 15,5 mg/kg and Zn level 229 mg/kg comparing with 0,01 mg/kg Cd, 0,8 mg/kg Pb and 3,1 mg/kg Zn in the clean area. In mellifera flora samples the metal levels were higher than in honeybees while in the honey samples the concentrations of heavy metals were higher than in unpolluted area but under the allowable limits.

The results indicated that heavy metals originating from immissions generated a persistent contamination which induced a high absorbtion of polluants into bee bodies during foraging. Honeybees can be considered representative bioindicators of environmental pollution.

Institute for Diagnosis and Animal Health, Department of Toxicology, Bucharest, Romania, E-mail: nica.daniela©idah.ro

In a morning the beekeeper of an individual farm observed a high mortality of his colonies. He found an excesive number of dead bees in the front of all hives. The apiary was situated in the backyard near a rape plantation. Because the damages were 100% from bees colonies he suspected that there were effectuated phytosanitary treatments to the plants. To establish the cause of the death he called the veterinarian who decided to sent immediately bees and rape samples for toxicological investigations.

Due to the placement of the hives and rapid death of the bees chemical analysis were first performed to identify fipronil (an usual pesticide for rape treatments in our country), organophosphorates, organochlorates and piretroids but the results were negative both for bees and plant samples. Then the analysis were done for another toxic compunds and it was chosen the identication of carbofuran. First step to determinate this compound was done using thin layer chromatography. Characteristic spots developed on the thin layer plate indicated the presence of carbofuran only in bees samples. The confirmation of carbofuran involved GC/MS technique which showed typical ions and retention time for this compound in bees and in spiked bees samples with carbofuran.

The death of the bees was an acute intoxication with carbofuran, a highly toxic compound for animals, birds, insects and fish.

Our results were sent to the beekeeper and to the police department to make investigations as to find of the suspect person.

Istituto Zooprofilattico Sperimentale delle Venezie, 35020 Legnaro (PD), Italy

The use of some products containing chlorfenvinfos, as Birlane® and Supona®, in the control of Varroa destructor infestation of honeybees is a diffuse practice among Italian beekeepers. The use of these products is in contrast with the Community legislation that not includes chlorfenvinfos as active substance in any authorized veterinary medicinal products intended for beekeeping (Regulation (CE) 2377/90). Furthermore, in Italy, any authorization of commercial products containing this active ingredient has been lifted to the end of 2003 (Regulation (CE) 2076/2002).

Actually Birlane® is easily available on illegal market and its use is much widespread, mainly due to its high efficacy against Varroa-mite. As occurs for other not authorized products, e.g. antibiotics, the use of chlorfenvinfos possesses great risks to Italian honey production. In fact, the presence of residues can cause, not only the withdrawal of the product from sale, but also a negative impact on the entire Italian beekeeping sector. Pharmacoresistance of Varroa-mite against organophosphates should also be considered.


Is the bacterial community of honey bee intestines affected by the consumption of insecticidal proteins?

D. Babendreier, D. Joller, J. Romeis, F. Bigler and F. Widmer

Agroscope Reckenholz-Tänikon Research Station ART, Reckenholzstr. 191, 8046 Zürich, Switzerland

Honey bees are important non-target species when the risks of insect resistant, transgenic plants are being assessed. Proteins expressed by these plants generally affect the intestinal tract of sensitive insects. We here tested whether the intestinal microflora of honey bees is affected by the consumption of transgenic pollen or insecticidal proteins and whether it is a good indicator for demonstrating sub-lethal effects on this non-target organism. We used a cultivation-independent approach based on PCR-amplified partial small subunit rRNA genes and genetic profiling by terminal restriction fragment length polymorphism (T-RFLP) to characterize the bacterial community of adult honeybee intestines. Bees were fed with Bt pollen (transgenic maize cultivar MON 810), Bt toxin (Cry1Ab) or the Kunitz Soybean trypsin inhibitor (SBTI) for the first ten days of their life. In addition, we also analyzed free-flying bees from two locations in Switzerland collected at two different times during the season. Neither the Bt pollen treatment nor high concentrations of Cry1Ab affected bacterial community in honeybee intestines significantly. In contrast, bees that were fed with high concentrations of SBTI had a significantly reduced number of bacteria in their midgut. However, in the field collected bees, changes in bacterial community were also found in relation to location or season. Since changes in the bacterial community were only observed at the high concentration of SBTI tested, which bees are unlikely to consume under natural conditions, we expect that these changes would not have detrimental effects on the health of honey bee colonies.

CNRS, E-mail: rortais©pge.cnrs-gif.fr

Systemic insecticides, which are applied as seed formula and thereafter permanently present in the pollen and nectar of plants present a risk to honeybees which collect and consume pollen and nectar during their entire life. However, the actual directives (EC-91/414) cannot assess properly the risk posed by systemic insecticides because it is based on HQ (Hazard Quotient), a value that does not take into account the application mode of these insecticides (i.e. not sprayed and not limited in time) and the biology of a honeybee colony (i.e. a superorganism). In order to overcome this lack, we propose a new risk assessment approach derived from various intoxication scenarios with the example of honeybees and imidacloprid, a systemic insecticide. This approach is based on the new and existing chemical substances directive which compares levels of exposure (PEC-Predicted Environmental Concentration) to levels of toxicity (PNEC-Predicted No Effect Concentration) of chemicals to organisms in order to protect ecosystems. PNECs are determined from the available data found in the literature on acute, chronic, and sublethal toxicities of imidacloprid to honeybees, to which specific assessment factors are applied. PECs are calculated with the known amounts of contaminated pollen and nectar (e.g. 1.9 ppb in sunflower nectar and 3.4 ppb in sunflower and maize pollens) consumed by different categories of honeybees. Results highlight a risk for all categories of honey bees, in particular for hive bees. New perspectives are given to better assess the risk posed by systemic insecticides to honey bees.

Tested methods for the use of honeybees as bioindicators

A. Rortais, Y. Loublier, L. Garnery , G. Arnold

CNRS, E-mail: rortais©pge.cnrs-gif.fr

The CNRS team is developing tested methods for the use of honeybees as bioindicators to detect and monitor (i) temporal and spatial changes in plant diversity and (ii) various sources of environmental pollution. This work is conducted within the consortium FRAGILE that includes several institutions (CNRS, Université de Versailles St Quentin-en-Yvelines, Université Paris XIII, Bergerie Nationale) which objective is to design proper indicators, based on a multi-disciplinary approach, to maintain sustainable use of biodiversity in the region of Ile-de-France (IDF), around Paris. To test these methods, behavioural (honeybees activity and dysfunction), palynologic, chemical (pollen dosages), genetics (populations heterozygosis, stress proteins) and morphometrics (fluctuating asymetry, malformations) measurements are conducted at two contrasted sites – an undisturbed area comprising forest and grasslands and - a disturbed agricultural location. Data is analysed with available expert systems, and methods are automated to generalise the use of honeybees as indicators of plant diversity and ecosystem pollution and dysfunction.


Consequences of a hive treatment with tetracycline on the quality of the honey

Anne-Claire Martel,Sarah Zeggane, Patrick Drajnudel, Jean-Paul Faucon and Michel Aubert

AFSSA Sophia Antipolis, E-mail: ac.martel©afssa.fr

Tetracyclines (TCs) are used to control bacterial diseases such as European and American foulbrood, which may cause severe losses in honeybee population and honey production. By using 24 hives randomly distributed into four groups of six hives, this study was performed to measure the occurrence of tetracycline hydrochloride (TC) residues in honey following two types of TC application. Two groups of colonies were treated 3 times with 0.5 g of TC in 1 litre of syrup (group S) or in 10 g of powdered sugar (group P). Six hives of a first control group (C) fed with untreated syrup were installed at 20 and 45 metres from groups S and P respectively. A second control group (DC) was set up 3 km away. Honey was sampled at different times from all hives and was analysed to follow the persistence and diffusion of TC residues into the apiary. One day after the last application, the mean TC concentration in brood chamber honey was ten times higher in group S (40.7 mg.kg-1) than in group P (4.34 mg.kg-1). After 146 days, the mean TC concentration in harvested honey was 1.54, 0.35, and 0.15 mg.kg-1 for groups S, P and C respectively. The control group C had been contaminated by drifting from treated hives. In all hives of group DC, no residues were detected at any time of the study. The half-life time of TC in honey from supers was similar in groups C, S and P and was equal to 65 days. These results show that the TC must be used with precaution in honey production.


Transfer and accumulation of heavy metals from cotaminated soils into the nests of Bombus terrestris L. (Hymenoptera, Apoidea) and little hives of Apis mellifera L.(Hymenoptera, Apoidea)

Andreé Hamm

INRES - Institute of Crop science and Ressource Conservation - Animal Ecology, E-mail: anham©web.de

We show the results of a studie, in wich we tried to find out, if the different components of the nests of Bombus terrestris and the hives of Apis mellifera can be used as a bioindikator for the heavy metal contamination of soils.


Death rate and weakening of honeybees in the south part of Belgium

Bach Kim Nguyen

Faculté Universitaire des sciences agronomiques de Gembloux - Unité d\'Entomologie fonctionnelle et évolutive, E-mail: nguyen.b©fsagx.ac.be

For several years, abnormal high death rate and weakening of honeybee (Apis mellifera L.) have been very often observed in the south part of Belgium (Walloon Region). Although two new insecticides, imidacloprid and fipronil, are regarded as being the single cause of the honeybee collapse, many multifactorial studies carried out in Europe and in North America clearly highlight many factors of risk: parasites, diseases, climate, food resources, low farmland biodiversity. The honeybees collapse stigmatizes a deep uneasiness at apiarian world confronted with a more and more anthropized environment.

Then, an exploratory multifactorial study was set up in order to discover the factors of risks which are likely to influence the vitality of the bees in the south part of Belgium. In this project, we study pesticides, pathologies and environment (nutritive resources, climates, etc.).

A list of pesticides including both product types used in apiculture and for the surrounding agriculture has been set up and quantification of residues of those pesticides (about 50 analytes) in honey, beeswax and bees were carried out.

The pathologies were studied combining field observations, microscopic, classic microbiological and molecular approaches.

Finally, the correlation between the honeybee colonies mortality and the presence of pesticides, pathologies and the environment were analyzed.

Sub-lethal effects of pesticides

Helen Thompson

CSL, York UK, E-mail: h.thompson©csl.gov.uk

Pesticides may affect honeybees by causing direct mortality or sub-lethal effects which may affect the individual and thus the colony. Effects at the level of the individual may include, for instance, decreased ability to return to the colony and may result in effects at the colony level such as reductions in colony strength. In the last few years, evaluation and assessment of possible sublethal effects of pesticides on bees has been increasingly discussed by scientists and regulatory authorities. This discussion has been enhanced by allegations, particularly in France, that exposure of honeybees to systemic insecticides may result in sublethal effects directly related to the reported colony losses. Novel study designs and data requirements have subsequently been proposed to test for sublethal effects on individual honeybees, in particular cases or even on a regular basis. This paper will consider whether and, if so, how sublethal effects should be incorporated into risk assessment by considering: What do we mean by a sublethal effect?

What sublethal effects should we measure, when and how?

How do we link sublethal effects to effects at colony level?

What should trigger assessment of sublethal effects?

How do we include sublethal effects in risk assessments?


Concentration of lead and cadmium in bees and bee bread

Anna Spodniewska, Konstanty Romaniuk

University of Warmia and Mazury in Olsztyn, Poland, E-mail: anspod©uwm.edu.pl

Studies were carried out in randomly selected apiaries of Warmia and Mazury province. At the turn of June and July the house bees and bee bread were collected from 5 colonies of each apiary in 22 communes of the province. Quantitative determination of lead (Pb) and cadmium (Cd) was done by the method of absorption atomic spectrophotometry.
The presence of lead and cadmium was found in the all samples of bees and bee bread . Lead (Pb) content in bees fluctuated between 0.234 mg/kg and 1.165 mg/kg and in bee bread between 0.074 mg/kg and 0.135 mg/kg however cadmium (Cd) was respectively 0.067-0.324 mg/kg and 0.030-0.072 mg/kg.

The obtained results indicate the low concentration of lead and cadmium in bee bread. The average content of Pb and Cd in bees from individual communes was about 1.7-12.7 and 1-10.8 times higher than in bee bread. The highest concentrations of Pb (0.932-1.421 mg/kg) and cadmium (0.210-0.324 mg/kg) were ascertained in bees from apiaries situated in central and western part of Warmia and Mazury province. In these communes, agricultural and food industry is concentrated, main transit roads run to the Russian border as well as manufacturing agriculture is developed.