Browsing by Person "Hasselmann, Martin"

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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.
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.
Classification and characterization of Ethiopian honey bees (Apis mellifera) based on morphometric, genetic and socio-economic analyses
(2022) Hailu, Teweldemedhn Gebretinsae; Hasselmann, Martin
Ethiopia is a major beekeeping country located in northeast Africa where several evolutionary lineages of Apis mellifera contact. A unique practice of honey bee colony marketing which involves broad agro-ecological zones (AEZs) is a developing trend in the northern part of the country such as Tigray region in association with apicultural development. Several studies based on classical morphometry on the Ethiopian honey bee subspecies classification debated from the unique Apis mellifera simensis to five others. Moreover, the genetic diversity, adaptation, gene flow and inter-relationships of the honey bees between AEZs were not disentangled – a challenge for planning sustainable apicultural development and conservation. Therefore, this study was conducted to elucidate the honey bees of Ethiopia in a context of apicultural transformation using integrated methods: morphometrics, genetics, colony market survey and metadata analyses on beekeeping development. The results of geometric morphometric analyses confirmed that Ethiopian honey bees represented by Apis mellifera simensis references belong to a separate lineage (Y) compared to A, O, M and C, and the present sample belonged to Y. This supported the hypothesis of five major honey bee lineages of the honey bee Apis mellifera. Similarly, a maximum likelihood phylogenetic tree analysis based on the mitochondrial COI-COII showed that most of the Ethiopian honey bees belong to lineage Y. However, a substantial proportion of the samples from the northern part of the country clustered with lineage O, which support the hypothesis that there is close contact between Y and O. Both geometric morphometry and classical morphometry differentiated the Ethiopian honey bees from all references including A. m. monticola, A. m. scutellata, A. m. jementica, A. m. adansonii but grouped with A. m. simensis. Genetically, five DraI haplotypes (COI-COII) were found to be randomly distributed across AEZs, indicating a substantial gene flow. Consequently, the level of genetic differentiation among the Ethiopian honey bee subpopulations defined by local areas and AEZs was generally low based on r7-frag nuclear marker, which is identified to be associated with adaptation to habitat elevation in East African honey bees. Similarly, nucleotide diversity consistently decreased with increasing elevation – indicating a reduced effective population size in the highlands. Results obtained from colony market survey showed that the honey bee swarms are reproduced in a few highlands and re-distributed throughout the region. Colony buyers have preferences of color and AEZ of origin of the honey bees, which led to a one-way flow and eroded the overall level of genetic differentiation. However, a marked differentiation was detected between the highland and lowland honey bees in relic communities where an allelic length polymorphism was observed as a signature of local adaptation. Altogether, Ethiopian honey bees belong to the lineage Y and subspecies A. m. simensis, and are characterized by a high level of gene flow enhanced by colony marketing; but a conserved signature of local adaptation to higher elevations was identified in less disturbed communities. Further studies based on genome-wide analyses and field experiments, focusing on undisturbed communities, can provide more insights into adaptation, admixture and management implications. Sustainable bee breeding and extension services that enable local beekeeping without colony trade and transportation will help to promote apiculture and genetic conservation.
European population genomic differentiation and dispersal pattern of the invasive beetle Anoplophora glabripennis
(2023) Häussermann, Iris Hanna; Hasselmann, Martin
Anthropogenic activities (e.g. homogenized habitats, trade) are the main factors to facilitate the increasing rates of invasive alien species. In this study, the invasion of the Asian long-horned beetle (Anoplophora glabripennis) was examined. Its native distribution is eastern Asia (China, Korean peninsula), but by extensive trade, this beetle was introduced via wood packing materials to North-America (1996) and Europe (2001). ALB attacks healthy broadleaved trees (e.g. Acer spp., Salix spp., Populus spp.), which can become lethal due to larval feeding. This study aims to detect genetic differences and kinship between the European infestation sites in Germany, Switzerland and Italy, from which the introduction and dispersal patterns can be deviated. Therefore, mitochondrial (mt) DNA-markers of the Cytochrome oxidase subunits I and II genes (COI and II) were used (ch. A and B, Sanger sequencing), as well as genome wide single nucleotide polymorphisms (SNPs), which were obtained by a Genotype-by-sequencing (GBS) approach (ch. C, Illumina sequencing). The results of this population genomic study of invasive European ALB populations showed very complex introduction patterns into Europe including multiple independent introductions characterized by the high population structure between the European infestation sites and some cases of human mediated secondary dispersal.
Mitochondrial haplotypes, gene expression and nuclear diversity in two strains of laying hens
(2021) Dreyling, Clara; Hasselmann, Martin
The domesticated chicken (Gallus gallus domesticus) is the most popular and widely spread domestic fowl worldwide, providing human with a stable source of protein in form of meat and eggs for centuries. The ongoing growth of human population increases the need for food and made poultry production one of the fasted growing sectors in the past decades. This need for food has resulted in several different strains which outperform their wild ancestors in terms of meat and egg production. During the past decades not only animal welfare gained importance but also ecological aspects such as global warming and the shortage of resources are becoming more important to society. One important resource for mankind which is becoming shortened is phosphorus (P), whose deposits in form of rock phosphate could be exhausted within the next 50-100 years. 90% of P supply is used in agriculture as fertilizer, whose demand will increase as well with growing population. This thesis focuses on the mitochondrial genetic background and mitochondrial related gene expression in the context of the productive life span and different diets in two contrasting high-yielding strains of laying hens, Lohmann Brown-Classic (LB) and Lohmann LSL-Classic (LSL). Mitochondria, which are commonly known as the powerhouse of the cell due to their role as the main producer of energy, play roles in other processes from cellular homeostasis to the process of ageing. The process of oxidative phosphorylation depends on the availability of P and thus, they become an important part of the complex framework of P utilization. In addition, mitochondrial haplotypes are known to affect physiological traits such as body weight in laying hens or important traits such as e.g. the metabolic capacity in dairy cows. It is known, that single mutations in the mitochondrial genome lead to a better adaptation to height in the Tibetan chicken or play a role in diseases from Alzheimer to obesity or lead to resistance to disease such as Marek’s disease in birds. This work provides insight into the whole mitochondrial genome of 180 laying hens of two commercial strains and links this information to physiological traits and genetic diversity. In addition, the first large-scaled gene expression analyses in the context of the productive life span and different P and Ca contents in laying hens is implemented. The analysis of mitochondrial haplotypes revealed a low level of genetic diversity with only three haplotypes within the LB strain while all LSL hens shared the same mitochondrial genome. Following from this observation, the nuclear genome was analysed based on genotyping data to reveal the whole genetic diversity of both strains. On the nuclear genetic level, both strains appeared as clearly distinct and equally diverse, while some individuals appear as strikingly close related. These individuals are mostly half-siblings sharing the same mitochondrial haplotype, underlining the need for more analyses about the genetic structure about the parental generation, especially the maternal background. Although there were no strong associations were found between the mitochondrial haplotypes and the analysed phenotypic traits (feed intake, body weight, P and Ca utilization), the differences between the strains indicate a potential involvement of the mitochondrial genetic background. The gene expression analyses revealed tissue type and point of the productive life span as the main influencers on gene expression while the influence of the strain is secondary. In addition, the expression of the gene GAPDH, which is frequently used as a reference gene for normalization in gene expression studies, was influenced by tissue and strain, leading to the decision to exclude it as a reference, that should be considered for in further studies. Further, no influence of the changes in dietary P and Ca on gene expression could be observed, suggesting that a reduction of 20% of both minerals is possible without the need to adapt gene expression. However, the results show, that a reduction of both minerals has less effect than a reduction of P alone, leading to an imbalance. In the context of the productive live span, mitochondrial and mitochondrial regulatory genes react contrary, illustrating the complexity of mitochondrial gene expression and regulation. In addition to the higher variance in the analysed phenotypic traits and mitochondrial genome in LB hens, they showed signs of increased oxidative stress compared to LSL hens. In the context of the productive life span, a potential higher demand for energy is suggested, since OXPHOS related gene expression is increasing. As a conclusion this work provides an insight into the mitochondrial genome and provides the first large scaled analysis of mitochondrial linked gene expression in two contrasting laying hen strains.
Reduced parasite burden in feral honeybee colonies
(2023) Kohl, Patrick L.; D'Alvise, Paul; Rutschmann, Benjamin; Roth, Sebastian; Remter, Felix; Steffan‐Dewenter, Ingolf; Hasselmann, Martin
Bee parasites are the main threat to apiculture and since many parasite taxa can spill over from honeybees (Apis mellifera) to other bee species, honeybee disease management is important for pollinator conservation in general. It is unknown whether honeybees that escaped from apiaries (i.e. feral colonies) benefit from natural parasite‐reducing mechanisms like swarming or suffer from high parasite pressure due to the lack of medical treatment. In the latter case, they could function as parasite reservoirs and pose a risk to the health of managed honeybees (spillback) and wild bees (spillover). We compared the occurrence of 18 microparasites among managed (N = 74) and feral (N = 64) honeybee colony samples from four regions in Germany using qPCR. We distinguished five colony types representing differences in colony age and management histories, two variables potentially modulating parasite prevalence. Besides strong regional variation in parasite communities, parasite burden was consistently lower in feral than in managed colonies. The overall number of detected parasite taxa per colony was 15% lower and Trypanosomatidae, chronic bee paralysis virus, and deformed wing viruses A and B were less prevalent and abundant in feral colonies than in managed colonies. Parasite burden was lowest in newly founded feral colonies, intermediate in overwintered feral colonies and managed nucleus colonies, and highest in overwintered managed colonies and hived swarms. Our study confirms the hypothesis that the natural mode of colony reproduction and dispersal by swarming temporally reduces parasite pressure in honeybees. We conclude that feral colonies are unlikely to contribute significantly to the spread of bee diseases. There is no conflict between the conservation of wild‐living honeybees and the management of diseases in apiculture.
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.