Landesanstalt für Bienenkunde

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    Honey bees (Apis mellifera) preselected for Varroa sensitive hygiene discriminate between live and dead Varroa destructor and inanimate objects
    (2023) Sprau, Lina; Traynor, Kirsten; Rosenkranz, Peter
    Varroa destructor is one of the main causes of colony losses of the western honey bee ( Apis mellifera ). Many efforts exist to breed honey bees resistant to V. destructor . Varroa sensitive hygiene (VSH) is a commonly selected behavioural trait; VSH workers remove the pupae of mite infested brood cells with high efficiency, interrupting the reproduction of the mite. The cues and triggers for this behaviour are not yet fully understood. To determine what elicits this removal behaviour, we examined preselected VSH workers´ responses to four different groups of objects inserted into freshly capped cells: live mites, dead mites, odour reduced mites, and glass beads. These were also compared to control cells that were opened and closed without inserting any object. The pupae in cells containing inorganic objects (glass beads) were removed at similar rates to the control, demonstrating that an object alone does not trigger a removal response. Dead and odour reduced mites were removed at a higher frequency than control cells, but less frequently than live mites. Workers sometimes removed items resting near the top of the cell without removing the pupa. Our results demonstrate that although mite odour from dead mites triggers removal behaviour, the pupa of cells containing live mites were removed more frequently, suggesting that other cues (i.e. odour from feeding wound) or signals (i.e. pupal movement to signal distress) are important. Future research should focus on elucidating these other cues or signals from the brood and mites, as mite presence alone seems to be insufficient.
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    Same data, different analysts: variation in effect sizes due to analytical decisions in ecology and evolutionary biology
    (2025) Gould, Elliot; Berauer, Bernd J.; Ernst, Ulrich Rainer; Zitomer, Rachel A.
    Although variation in effect sizes and predicted values among studies of similar phenomena is inevitable, such variation far exceeds what might be produced by sampling error alone. One possible explanation for variation among results is differences among researchers in the decisions they make regarding statistical analyses. A growing array of studies has explored this analytical variability in different fields and has found substantial variability among results despite analysts having the same data and research question. Many of these studies have been in the social sciences, but one small “many analyst” study found similar variability in ecology. We expanded the scope of this prior work by implementing a large-scale empirical exploration of the variation in effect sizes and model predictions generated by the analytical decisions of different researchers in ecology and evolutionary biology. We used two unpublished datasets, one from evolutionary ecology (blue tit, Cyanistes caeruleus , to compare sibling number and nestling growth) and one from conservation ecology ( Eucalyptus , to compare grass cover and tree seedling recruitment). The project leaders recruited 174 analyst teams, comprising 246 analysts, to investigate the answers to prespecified research questions. Analyses conducted by these teams yielded 141 usable effects (compatible with our meta-analyses and with all necessary information provided) for the blue tit dataset, and 85 usable effects for the Eucalyptus dataset. We found substantial heterogeneity among results for both datasets, although the patterns of variation differed between them. For the blue tit analyses, the average effect was convincingly negative, with less growth for nestlings living with more siblings, but there was near continuous variation in effect size from large negative effects to effects near zero, and even effects crossing the traditional threshold of statistical significance in the opposite direction. In contrast, the average relationship between grass cover and Eucalyptus seedling number was only slightly negative and not convincingly different from zero, and most effects ranged from weakly negative to weakly positive, with about a third of effects crossing the traditional threshold of significance in one direction or the other. However, there were also several striking outliers in the Eucalyptus dataset, with effects far from zero. For both datasets, we found substantial variation in the variable selection and random effects structures among analyses, as well as in the ratings of the analytical methods by peer reviewers, but we found no strong relationship between any of these and deviation from the meta-analytic mean. In other words, analyses with results that were far from the mean were no more or less likely to have dissimilar variable sets, use random effects in their models, or receive poor peer reviews than those analyses that found results that were close to the mean. The existence of substantial variability among analysis outcomes raises important questions about how ecologists and evolutionary biologists should interpret published results, and how they should conduct analyses in the future.
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    Digest: Better constructing the future than choosing a new habitat
    (2022) Ernst, Ulrich R.
    What strategy should an individual follow in a heterogeneous environment when its phenotype is not optimized for its current environment: make changes to the environment (habitat construction), move to a different place (habitat choice), or both? Scheiner et al. used an individual‐based model to investigate the interaction of habitat choice and habitat construction. In most situations, habitat construction was superior to either habitat selection or a mixed strategy.
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    Wirksamkeit, Nebenwirkung und Verteilung von Lithiumchlorid: ein neuer Wirkstoff zur Behandlung von Varroa destructor bei Honigbienenvölkern (Apis mellifera)
    (2024) Rein, Carolin Vanessa; Rosenkranz, Peter
    Honigbienen sind unverzichtbare Bestäuber sowohl für unser Ökosystem als auch für die Landwirtschaft. Die weltweit verbreitete ektoparasitische Milbe Varroa destructor stellt seit Jahrzehnten das größte Problem für Honigbienen und die Imkerei dar und gilt zweifellos als Hauptverursacher der periodischen Völkerverluste (Genersch et al. 2010; Le Conte et al. 2010). Derzeit gibt es kein zufriedenstellendes Behandlungsverfahren, das alle Anforderungen der Imker erfüllt. Im Jahr 2018 wurde mit Lithiumchlorid (LiCl) ein neuer Wirkstoff mit varroazider Wirkung entdeckt, welcher eine sehr gute Wirksamkeit auf Varroamilben mit einer guten Verträglichkeit für adulte Bienen verbindet (Ziegelmann et al. 2018). Aufgrund der systemischen Wirkungsweise kann er zudem sehr einfach angewendet werden und bietet dadurch großes Potential für eine effektive Varroabehandlung. Im Rahmen der Dissertation wurden verschiedene Applikationsformen von LiCl zur Behandlung von Bienenvölkern getestet und Daten zur Wirksamkeit, Nebenwirkung und Verteilung des Wirkstoffes erhoben. Die gesammelten Daten sollen dazu dienen, eine fundierte Bewertung der Chancen und Risiken für ein Zulassungsverfahren zu ermöglichen und erfolgsversprechende Strategien für die Zulassung zu entwickeln. Zu diesem Zweck wurden verschiedene Feld- und Laborexperimente durchgeführt. Unter praxisnahen Freilandbedingungen führt die Applikation von LiCl zu konzentrationsabhängigen Schäden und hohen Ausräumraten der Bienenbrut. Diese Brutschäden sind von der Dauer der Fütterung und dem Larvenalter abhängig (Rein et al. 2022). Bei einer Fütterung von 25 mM LiCl überleben nur knapp 40% der Bienenbrut. Analysen der Lithium-Konzentration verschiedener Larvenstadien zeigen, dass in zwei Tage alten Larven noch kein Lithium nachgewiesen werden kann. Die Analysen der Futtersaftdrüsen haben bestätigt, dass das Sekret der Ammenbienen, mit dem die Königin und die jungen Larvenstadien gefüttert werden, weitgehend frei von Lithium ist (Rein et al. 2024). Daher sollte eine LiCl-Applikation kein Risiko für die Königin, oder die jungen Larven darstellen. In älteren Larven steigt die Lithium-Konzentration dagegen signifikant an (Rein et al. 2022). Dies ist auf die stadienspezifische Futterumstellung von reinem Futtersaft auf gemischtes Larvenfutter zurückzuführen. Hierbei wird auch eingelagertes Futter aus den umliegenden Waben beigemengt, wodurch auch zuvor eingelagertes Lithium weitergegeben werden kann. Aufgrund der schädigenden Wirkung von LiCl auf die Bienenbrut wurde eine Applikation für brutfreie Völker entwickelt und die Wirksamkeit unter Feldbedingungen getestet. Die Kombination einer brutfreien Phase durch Sperren der Königin mit der LiCl-Behandlung erzielte durchschnittliche Wirkungsgrade von 78 – 98% (Rein et al. 2024). Die höchsten Wirkungsgrade wurden erst bei einer Fütterungsdauer von > 5 Tagen erreicht, was zu Schäden bei der neu angelegten Brut nach der Brutpause führte. In Käfigversuchen, in denen jeweils eine Biene mit einer parasitierenden Milbe mit LiCl gefüttert wurde, konnten dagegen bereits nach 48 Stunden Milbenmortalitäten von über 95% erreicht werden (Rein et al. 2024). Offenbar verzögerte sich im Volk der Wirkungseintritt aufgrund des sozialen Futteraustauschs (Trophallaxis) und einem damit verbundenen Verdünnungseffekt des Wirkstoffs. Jedoch ist dieser Futteraustausch notwendig, um den Wirkstoff gleichmäßig im Volk zu verteilen und alle Milben zu erreichen. Es besteht daher weiterer Optimierungsbedarf, um die für die Zulassung notwendigen Wirkungsgrade von über 90% zu erreichen, ohne die Bienenbrut zu schädigen. Anhand von quantitativen Lithium-Analysen über ICP-OES konnte bestätigt werden, dass der Wirkstoff im Volk gleichmäßig verteilt wird. Bereits einen Tag nach der LiCl-Applikation wurden im Schnitt 93 mg/kg Lithium in den Honigblasen der Bienen nachgewiesen, unabhängig von ihrer Position im Volk (Rein et al. 2024). Analysen der Bienen-Hämolymphe zeigen, dass bereits nach einer 12-stündigen Fütterung der Bienen im Käfig ein Gleichgewicht von 5 – 8 mg/kg Lithium erreicht wird, welches zum Tod der Milbe führt (Rein et al. 2024). Das Lithium sollte daher nach einer LiCl-Applikation möglichst schnell im gesamten Volk verteilt werden und ausreichend lange im Volk zirkulieren, um möglichst in allen Bienen die für Milben tödliche Konzentration zu erreichen. Diese Konzentration sollte für etwa 48 Stunden aufrechterhalten werden, um eine Milbenmortalität von über 95% zu erreichen. Die im Rahmen der vorliegenden Dissertation durchgeführten Versuche bieten eine umfangreiche Basis für die Entwicklung einer zulassungsrelevanten Applikation. Es konnte gezeigt werden, dass Lithium eine hohe akarizide Wirkung besitzt und lediglich noch einige Optimierungen notwendig sind, um eine bessere Verteilung des Wirkstoffes und damit stabile Wirkungsgrade von über 90% bei brutfreien Völkern zu erreichen. Für brütende Völker müssen jedoch alternative Behandlungsmöglichkeiten entwickelt werden, um den Kontakt der Brut mit dem Wirkstoff zu vermeiden.
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    Temporal increase of Varroa mites in trap frames used for drone brood removal during the honey bee season
    (2022) Odemer, Richard; Odemer, Franziska; Liebig, Gerhard; de Craigher, Doris
    Varroa mites are highly attracted to drone brood of honey bees (Apis mellifera), as it increases their chance of successful reproduction. Therefore, drone brood removal with trap frames is common practice among beekeepers in Europe and part of sustainable varroa control. However, it is considered labour‐intensive, and there are doubts about the effectiveness of this measure. At present, it is mostly unknown how many mites a drone frame can carry at different times of the season, and how many mites can be removed on average if this measure is performed frequently. Therefore, we sampled a total of 262 drone frames with varying proportion of capped cells (5–100%) from 18 different apiaries. Mites were washed out from brood collected from mid‐April to mid‐July based on a standard method to obtain comparable results. We found that a drone frame carried a median of 71.5 mites, and with the removal of four trap frames, about 286 mites can be removed per colony and season. In addition, mite counts were significantly higher in June and July than in April and May (Tukey‐HSD, P < 0.05). The number of mites and the proportion of capped cells, however, were not correlated (R2 < 0.01, P < 0.05). Our results suggest that drone brood removal is effective in reducing Varroa destructor numbers in colonies, supporting the findings of previous studies on the efficacy of this measure. Although mite counts varied, we believe that increasing sample size over different seasons and locations could elucidate infestation patterns in drone brood and ultimately improve drone brood removal as an integrated pest management tool for a wider audience of beekeepers.
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    The emergence of ecotypes in a parasitoid wasp: a case of incipient sympatric speciation in Hymenoptera?
    (2021) Malec, Pawel; Weber, Justus; Böhmer, Robin; Fiebig, Marc; Meinert, Denise; Rein, Carolin; Reinisch, Ronja; Henrich, Maik; Polyvas, Viktoria; Pollmann, Marie; von Berg, Lea; König, Christian; Steidle, Johannes L. M.
    Background: To understand which reproductive barriers initiate speciation is a major question in evolutionary research. Despite their high species numbers and specific biology, there are only few studies on speciation in Hymenoptera. This study aims to identify very early reproductive barriers in a local, sympatric population of Nasonia vitripennis (Walker 1836), a hymenopterous parasitoid of fly pupae. We studied ecological barriers, sexual barriers, and the reduction in F1-female offspring as a postmating barrier, as well as the population structure using microsatellites. Results: We found considerable inbreeding within female strains and a population structure with either three or five subpopulation clusters defined by microsatellites. In addition, there are two ecotypes, one parasitizing fly pupae in bird nests and the other on carrion. The nest ecotype is mainly formed from one of the microsatellite clusters, the two or four remaining microsatellite clusters form the carrion ecotype. There was slight sexual isolation and a reduction in F1-female offspring between inbreeding strains from the same microsatellite clusters and the same ecotypes. Strains from different microsatellite clusters are separated by a reduction in F1-female offspring. Ecotypes are separated only by ecological barriers. Conclusions: This is the first demonstration of very early reproductive barriers within a sympatric population of Hymenoptera. It demonstrates that sexual and premating barriers can precede ecological separation. This indicates the complexity of ecotype formation and highlights the general need for more studies within homogenous populations for the identification of the earliest barriers in the speciation process.
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    Auslöser und Ausprägung des Varroa Sensitiven Hygiene (VSH) Verhaltens im Zusammenhang mit der Reproduktion der Varroamilbe Varroa destructor
    (2024) Fölsch, Lina; Hasselmann, Martin
    The ectoparasite Varroa destructor causes massive economic damages to the Western honey bee (Apis mellifera) and thus threatens the health of honey bee colonies worldwide. When left untreated, varroa populations can lead to high winter colony losses (Genersch et al. 2010; Traynor et al. 2020; Le Conte et al. 2010). Beekeepers must thus routinely treat against varroa to keep their colonies healthy. A sustainable solution to overcome the varroa problem is selection for varroa resistant honey bees. In addition to studies on natural selection against varroa, there is much interest in breeding varroa resistant honey bees. During varroa resistant selection the focus is often on different selection criteria, such as mite non-reproduction (MNR) and varroa sensitive hygiene (VSH). The selection criteria MNR describes the proportion of non-reproductive mites in a colony (Virag et al. 2022). VSH encompasses the specialized removal behavior of mite infested brood by worker bees (Dietemann et al. 2013; Harbo und Harris 2005). This brood removal interrupts varroa reproduction, which leads to reduced varroa infestation in honey bee colonies. The experiments in thesis were conducted as part of the SETBie project. The SETBie (Selection and Establishment of Varroa-tolerant Bee Colonies) project was a four-year selection project whose objectives included breeding a varroa-resistant honey bee in Baden-Württemberg, Germany. Several institutions and beekeeper organizations cooperated in this project. This dissertation focuses on the factors that trigger VSH behaviour, in addition to documenting the reproductive success of the mite (MNR) in selected colonies. Initial experiments investigated the relationship between VSH and the reproductive success of female Varroa (MNR). A widely held hypothesis was that the reproduction of the mite and the associated nymphal stages in the brood cells triggered the removal behaviour of the workers. However, this hypothesis was disproved by placing mites in capped brood cells and comparing the removal behaviour of reproducing mites with that of mites whose reproduction had been blocked by a special procedure (publication 1). Both groups of mites were removed at a rate of about 40% and did not differ significantly from each other. Therefore, reproduction of the mite could be excluded as a trigger for removal behavior of the workers. This result was supported by simultaneous analysis of VSH behaviour and the proportion of non-reproductive mites (MNR) in a large number of pre-selected colonies. It was also assumed that the increased number of non-reproductive mites (MNR) within a colony is directly connected to VSH behaviour, because the workers would primarily remove the reproductive mites, leaving only non-reproductive mites behind. In addition to providing evidence that mite reproduction is not the trigger of removal behaviour, our analysis of the data collected during the SETBie project showed no correlation between MNR and VSH values (publication 3). The MNR value of a colony therefore does not allow any conclusions to be drawn about the VSH value, which should be taken into account in recommendations for breeding practices. Furthermore, an experiment was conducted to rule out the possibility that removal behaviour effects the normal reproduction of the mite in the next cycle (publication 3). The need for other triggering factors for removal behaviour to occur was clearly demonstrated in this first set of experiments. Varroa mites manipulated to remove specific factors and inanimate objects were placed in capped brood cells to determine what influences removal behaviour. We found that an object (bead) alone does not induce removal behaviour (publication 2). Dead and odour-reduced mites were removed more frequently than the bead and control, but not anywhere near as frequently as live mites. We can thus assume that, in addition to the odour of the mite, the movement of the mite in particular has a signalling effect that triggers this removal behaviour. Other as yet unknown factors such as the reaction of the larvae to the parasitism are likely to play an important role in the removal behaviour of workers. The SETBie project took place over a period of four years, which allowed us to analyse colonies over several generations. Although MNR and VSH are in principle selectable traits, our analysis has shown that they are difficult to improve over multiple generations (publication 4). Colonies with low MNR values could often be improved by crossing them with a drone (artificial insemination) from a colony with a high MNR value. However, it also became clear that it is difficult to maintain high MNR values. Methodological difficulties in targeted selection of MNR/VSH also became noticeable during the collaborative project, which involved the cooperation of numerous breeders. Despite prior infestation of the selected colonies with 180 mites, it was often not possible to collect enough mites to make a valid statement about the reproductive capability of the mite. Even after opening many cells, the minimum number of 10 single infested cells could not be found, which means that MNR values could not be determined for about 63 % of the evaluated colonies due to the presence of too few mites. The selection and breeding work involved in this project is extremely labour-intensive and therefore expensive and requires a high level of expertise among the breeders. It was shown that MNR and VSH are in principle selectable and thus theoretically a Varroa-resistant honey bee line can be bred. However, for future projects, methodological problems need to be resolved or the evaluation methods of the MNR and VSH traits need to be simplified. An optimal solution would be the identification of molecular markers to support selection via genetic heritability. However, this requires further fundamental knowledge about the Varroa mite, the mechanisms that trigger the behaviour of the currently used selection traits, and more precise phenotypic definition of the resistant behaviours.
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    Zierpflanzen als Nahrungsquelle und Bewertung der Blütenmerkmale für die Attraktivität der Bestäuberinsekten im urbanen Raum
    (2022) Marquardt, Melanie; Rosenkranz, Peter
    Currently, an increasing extension of urban areas can be observed worldwide, which implies a concurrent loss in natural habitats. If the current biodiversity shall be preserved, efforts must be reinforced in order to provide alternative habitats for the flora and fauna in urban areas. Regarding the assessment of the ecological value of the habitat ‘city’, scientific studies came to varying conclusions. However, all of them emphasize the importance of green urban areas in promoting the urban biodiversity. It has been frequently pointed out that sufficient and suitable foraging resources are an important and basic requirement for the survival of flower-visiting insects. However, it has hardly been investigated whether ornamental plants are suitable foraging resources for insect pollinators. While this is still highly contentious, there is growing evidence that ornamental plants could contribute to the provision of pollen and nectar. But apparently, the attractivity of different ornamental plants varies widely and furthermore, not all pollinator groups can profit equally from the mostly exotic ornamental plants. At present, scientific data for all ornamental genera or even species are not available. So, the first aim of this study was the comparison of the attractivity of certain ornamental plants, in particular those with a high market value. In order to conduct such trials, already existing acquisition methods have been assessed and refined. A further and so far rarely considered focus of this thesis is the analysis of impact factors that might affect the composition and abundance of urban pollinators. In order to identify the pollinator friendliness of ornamental plants, field tests in urban areas and semi-field tests in flight tents were conducted during the years 2017 – 2019. In the first trail, raised flower beds with an identical set of ornamental plants were installed at 13 different locations in the city area of Stuttgart. During the summer months of the years 2017 and 2018, all flower beds were visited in weekly intervals. Over a time period of 20 minutes, the number of foraging insects – divided into different groups of insect pollinators – was recorded. In total, 10,565 nectar and/or pollen foraging insects were counted. First of all, this confirms that our selection of ornamental plants was used as a foraging resource by pollinating insects. The attractivity of the tested ornamental plants, however, varied to a considerable degree among the plant species and the number of counts ranged from 1.2 flower visits in 20 minutes on Bracteantha bracteata (strawflower) to 5.3 flower visits on Bidens (beggar-ticks). It is noteworthy, that the attractivity also varied within the cultivars of the same species, partly even to a greater extent than between species. Interestingly, not only the abundance but also the composition of pollinators varied among the different test plants. Furthermore, the applied statistical models indicate significant impacts of the study year and the location on the results. This highlights the need of a continuously testing of all ornamental plants in regard to insect friendliness, for which the described methods were found to be very appropriate (publication I). Pollinating insects often use characteristic floral traits of the plants for their decision to visit the respective flower. These floral traits are often genera-, species- or even cultivar-specific and have been well studied in the native plants. In contrast, very little is known about the role of floral traits in ornamental plants. This includes e. g. the petal colour, several floral morphomet-rics or the floral scent of different plants. The impact of these traits on attractivity for insects was analysed in semi-field experiments using Calibrachoa cultivars and Bombus terrestris as model pollinator. Similar to the first part of the thesis, the attractivity of the different cultivars varied significantly. While the floral scent explained the observed differences in attractivity only to a small extend, it could be shown unequivocally that the petal colour constitutes a significant factor in the attractivity on B. terrestris. For a better understanding on the impact of certain floral traits for pollinating insects, however, there is further research required (publication II).
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    Charakterisierung der Qualität von Blütenpollen in unterschiedlichen Regionen Baden-Württembergs
    (2022) Friedle, Carolin Gertrud Maria; Hasselmann, Martin
    Honey bees (Apis mellifera) collect nectar and pollen from plants to feed their brood. Pollen provides a wide range of nutrients, such as proteins and lipids, but also carbohydrates, vitamins and enzymes. Because of these ingredients, pollen is also attractive to humans and is used as a dietary supplement. However, honey bees collect pollen not only from wild plants, but also from flowering crops grown in agriculture. Accordingly, contamination from plant protection products can be found in bee pollen and bee bread. In order to get a deeper insight into the occurrence and distribution of pesticide residues during an entire season, a total of 102 daily pollen samples were collected from April to July 2018 using pollen traps in an orchard in southern Germany. Almost 90% of the pollen samples showed detectable levels of pesticide residues. A total of 29 pesticides were detected in the samples, with more than half being fungicides, followed by insecticides and herbicides. Maximum concentrations of up to 4500 ng/g could be measured at the end of April. Samples collected in early May and late June also showed high levels of pesticides. A general risk management was performed to assess the risk of the detected pesticide concentrations for honey bees. The microbial quality of bee pollen is highly dependent on its botanical and geographic origin, as well as climatic conditions and post-harvest processing steps by the beekeeper. If no processing steps such as freezing or drying follow after harvest, the growth of microorganisms can be promoted and the pollen quality can be influenced by negative side effects such as fermentation or the production of mycotoxins. Bacterial and fungal colonies can be determined both by culture-dependent methods such as colony counting on plates and by culture-independent methods such as 16-rRNA amplicon sequencing. Following the hypothesis that storage conditions influence the composition of microorganisms in bee pollen, freshly harvested bee pollen was stored for seven days in June 2018 and 2019 under defined conditions (cold, room temperature, warm) and analyzed by sequencing 16S and 18S PCR amplicons. The bacterial community varied slightly between the sites studied and showed no significant difference between the storage conditions. The fungal community showed significant differences both between the studied sites and between the different storage conditions. The dominant fungal genera in the pollen samples were Cladosporium, Aspergillus and Zygosaccharomyces. While Cladosporium was most dominant in freshly collected pollen and the percentage decreased during storage, Aspergillus and Zygosaccharomyces showed a significant increase especially under warm storage conditions. Other contaminants naturally produced by plants can also have negative impacts on human health. Pyrrolizidine alkaloids belong to a group of phytochemicals, of which more than 600 structures are known in around 3% of all flowering plants worldwide. PA are known to be able to cause both acute poisoning and chronic damage or cancer in animals and humans. In July 2019, pollen was collected at 57 locations in Baden-Württemberg and analyzed for 42 different PAs and their N-oxides in order to expand knowledge about PA contamination in pollen and to be able to estimate the risk of the concentrations. A total of 22 different PAs were detected in over 90% of all samples examined. Only 5% of the PA were obtained as PA from plants of Senecio sp. identified, while 95% of PAs with a botanical background are from Echium sp. and Eupatorium sp. could be identified. The maximum total concentration of PA per sample was determined to be 48,400 ng/g. According to the risk values calculated by the BfR, however, 42% of the samples represented an increased risk to human health.
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    The production of melezitose in honeydew and its impact on honey bees (Apis mellifera L.)
    (2021) Seeburger, Victoria; Hasselmann, Martin
    Honeydew honey is a honey type which is of high economic importance in Europe. Phloem sap feeding insects of the order Hemiptera (true bugs) excrete honeydew, the key component of honeydew honey. Beekeepers move their hives between forest regions so that their bees can process the honeydew into honey. In case of high osmolality in the phloem sap of the hemipterans’ host trees, they counteract osmotic pressure by osmoregulation and produce oligosaccharides such as melezitose. Melezitose-rich honeydew honey is a major issue for beekeepers; it crystallises and obstructs the combs, leading to an economical loss. Nevertheless, precise analyses of the conditions of the occurrence of melezitose have not been realised. Furthermore, it is not known which impacts the trisaccharide has on honey bee health and the honeydew flow disease documented in beekeepers’ journals can have one explanation in the nutrition on melezitose. In order to determine influence factors for the emergence of melezitose, more than 600 honeydew droplets from defined honeydew producer species were collected under different environmental conditions (hemipteran species (host tree specific), natural area, air temperature, relative humidity, altitude, time of the year and of the day) between 2016 and 2019. The sugar spectra were analysed by high performance anion exchange chromatography with pulsed amperometric detection. To obtain the impact of melezitose on honey bee health, additional feeding experiments with daily evaluation of food uptake, gut-body weight ratio and mortality have been realised between 2017 and 2019. Additionally, comprehensive 16S rRNA Illumina sequencing of the gut microbial community has been performed. Remarkable differences could be found in the amount of melezitose between honeydew samples collected from different honeydew producer species and according to different environmental conditions. Air temperature increases and decreases in relative humidity increased the melezitose production in honeydew by the observed seven hemipteran species. Both, scale insect species on Picea abies and aphid species on Abies alba produced significantly less honeydew containing melezitose than aphid species on Picea abies. Additionally, honeydew with increased melezitose content was significantly more frequent collected in natural areas with limited water reservoir capacities, at higher altitudes and years with low precipitation. All results lead to the conclusion that hemipteran species produce more melezitose when the host trees have less access to water, increasing the osmolality of the phloem sap and indirectly enhancing the osmoregulation with producing melezitose by hemipteran species. Bees fed with melezitose showed increased food uptake and higher gut-body weight ratio than the control groups. Furthermore, melezitose feeding caused disease symptoms such as swollen abdomen, abdomen tipping and impaired movement and a significantly higher mortality than in control groups. Gut microbiota analyses indicated a shift of the bacterial species Lactobacillus Firm-4 and Lactobacillus kunkeei in favour of Lactobacillus Firm-5 in melezitose fed bees. This PhD project provides the important knowledge about the indicators that point out an enhanced melezitose production. This is a valuable contribution to design a warning system for beekeepers that will help to prevent harmful nutrition for honey bees or crystallised honey in the future by timely removal of bee colonies from local regions at risk. Additionally, feeding experiments point out the high effort that is required for the degradation process of the large-molecule melezitose. This effort might lead to a higher uptake of food, heavier guts, shorter lifespan and a higher susceptibility to intestinal diseases. Finally, an evidence was presented that the lactic acid bacteria of the gut microbiota are significantly involved in the digestion of melezitose.
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