Honeybees are important pollinators of many crops and contribute to biological biodiversity. For years, a decline in bee populations has been observed in certain areas. This decline in honeybees is accompanied by a decrease in pollinator services. One factor contributing to the decline of bee colonies is the exposure to pesticides. Pesticide exposure of bees, among other effects, can negatively affect orientation, memory, immune system function and gene expression. Among the altered expressed genes are transcripts of endocrine regulation and oxidative phosphorylation. Endocrine regulation plays an important role in the development of nurse bees into foragers and oxidative phosphorylation is involved in energy metabolism. Most of these transcriptional changes were investigated using mixed aged honeybees derived from the same colony. Experiments using nurse bees or foragers of the same age but from different colonies are rare. In the present study, effects of the two pesticides chlorpyrifos and pyraclostrobin on the expression of transcripts linked to endocrine regulation and oxidative phosphorylation in foragers of the same age from three different colonies are investigated to fill this gap. These two pesticides were selected because negative effects at sublethal concentrations on bees are known and because they are found in pollen and nectar of crops and wild plants. For this purpose, 20–22 days old foragers of three different colonies were exposed to different sublethal concentrations of the selected fungicides for 24 h, followed by analysis of the expression of buffy, vitellogenin, hbg-3, ilp-1, mrjp1, 2 and 3, cox5a, cox5b and cox17. Some significant changes in gene expression of both endocrine regulation transcripts and oxidative phosphorylation were shown. Furthermore, it became clear that forager bees from different colonies react differently. This is especially important in relation to the risk analysis of pesticides. In addition, it could be shown that the expression of hbg-3 in the brain of bees is a robust marker to distinguish nurse bees from foragers at the molecular biological level. In summary, this study clearly shows that pesticides, which are often detected in pollen and nectar, display negative effects at sublethal concentrations on bees and that it is important to use bees from different colonies for risk assessment of pesticides.

Intensive growth of cyanobacteria in freshwater promoted by eutrophication can lead to release of toxic secondary metabolites that may harm aquatic organisms and humans. The serine protease inhibitor aeruginosin 828A was isolated from a microcystin-deficient Planktothrix strain. We assessed potential molecular effects of aeruginosin 828A in comparison to another cyanobacterial serine protease inhibitor, cyanopeptolin 1020, in human hepatoma cell line Huh7, in zebrafish embryos and liver organ cultures. Aeruginosin 828A and cyanopeptolin 1020 promoted anti-inflammatory activity, as indicated by transcriptional down-regulation of interleukin 8 and tumor necrosis factor α in stimulated cells at concentrations of 50 and 100 µmol·L⁻¹ aeruginosin 828A, and 100 µmol·L⁻¹ cyanopeptolin 1020. Aeruginosin 828A induced the expression of CYP1A in Huh7 cells but did not affect enzyme activity. Furthermore, hatched zebrafish embryos and zebrafish liver organ cultures were exposed to aeruginosin 828A. The transcriptional responses were compared to those of cyanopeptolin 1020 and microcystin-LR. Aeruginosin 828A had only minimal effects on endoplasmic reticulum stress. In comparison to cyanopeptolin 1020 our data indicate that transcriptional effects of aeruginosin 828A in zebrafish are very minor. The data further demonstrate that pathways that are influenced by microcystin-LR are not affected by aeruginosin 828A.

Neonicotinoids as thiamethoxam and thiacloprid are suspected to be implicated in the decline of honey bee populations. As nicotinic acetylcholine receptor agonists, they disturb acetylcholine receptor signaling in insects, leading to neurotoxicity and are therefore globally used as insecticides. Several behavioral studies have shown links between neonicotinoid exposure of bees and adverse effects on foraging activity, homing flight performance and reproduction, but the molecular aspects underlying these effects are not well-understood. In the last years, several studies through us and others showed the effects of exposure to neonicotinoids on gene expression in the brain of honey bees. Transcripts of acetylcholine receptors, hormonal regulation, stress markers, detoxification enzymes, immune system related genes and transcripts of the energy metabolism were altered after neonicotinoid exposure. To elucidate the link between homing flight performance and shifts in gene expression in the brain of honey bees after neonicotinoid exposure, we combined homing flight activity experiments applying RFID technology and gene expression analysis. We analyzed the expression of endocrine factors, stress genes, detoxification enzymes and genes linked to energy metabolism in forager bees after homing flight experiments. Three different experiments (experiment I: pilot study; experiment II: “worst-case” study and experiment III: laboratory study) were performed. In a pilot study, we wanted to investigate if we could see differences in gene expression between controls and exposed bees (experiment I). This first study was followed by a so-called “worst-case” study (experiment II), where we investigated mainly differences in the expression of transcripts linked to energy metabolism between fast and slow returning foragers. We found a correlation between homing flight duration and the expression of cytochrome c oxidase subunit 5A, one transcript linked to oxidative phosphorylation. In the third experiment (experiment III), foragers were exposed in the laboratory to 1 ng/bee thiamethoxam and 8 ng/bee thiacloprid followed by gene expression analysis without a subsequent flight experiment. We could partially confirm the induction of cytochrome c oxidase subunit 5A, which we detected in experiment II. In addition, we analyzed the effect of the feeding mode (group feeding vs. single bee feeding) on data scattering and demonstrated that single bee feeding is superior to group feeding as it significantly reduces variability in gene expression. Based on the data, we thus hypothesize that the disruption of energy metabolism may be one reason for a prolongation of homing flight duration in neonicotinoid treated bees.

The increased reports of wild bee declines and annual losses of managed bees pose a significant threat to biodiversity and agricultural productivity. While these losses and declines are likely driven by various factors, the exposure of bees to agrochemicals has raised significant concern due to their ubiquitous use and potential adverse effects. Despite numerous studies suggesting neonicotinoids can negatively affect bees at the behavioral and molecular level, data linking these two factors remains sparse. Here we provide data on the impact of an acute dose of the neonicotinoid thiamethoxam on the flight performance and molecular transcription profiles of foraging honey bees (Apis mellifera). Using a controlled experimental design with tethered flight mills, we measured flight distance, duration, and speed, alongside the expression of genes involved in energy metabolism, hormone regulation, and biosynthesis. Acute thiamethoxam exposure resulted in hyperactive flight behavior but led to significant dysregulation of genes associated with oxidative phosphorylation, indicating potential disruptions in cellular energy production. These molecular changes were particularly evident when bees engaged in flight activities, suggesting that the combined stress of pesticide exposure and physical exertion exacerbates negative outcomes. Our study provides new insights into the molecular mechanisms underlying neonicotinoid-induced impairments in bee physiology that can help understand the potential long-term consequences of xenobiotic exposure on the foraging abilities of bees and ultimately fitness.

Bees are often exposed to pesticides affecting physiological functions and molecular mechanisms. Studies showed a potential link between altered expression of energy metabolism related transcripts and increased homing flight time of foragers exposed to pesticides. In this study, we investigated the effects of thiamethoxam and pyraclostrobin on longevity, flight behavior, and expression of transcripts involved in endocrine regulation (hbg-3, buffy, vitellogenin) and energy metabolism (cox5a, cox5b, cox17) using radio frequency identification (RFID) technology and quantitative polymerase chain reaction. Parallel, a laboratory study was conducted investigating whether pesticide exposure alone without the influence of flight activity caused similar expression patterns as in the RFID experiment. No significant effect on survival, homing flight duration, or return rate of exposed bees was detected. The overall time foragers spent outside the hive was significantly reduced post-exposure. Irrespective of the treatment group, a correlation was observed between cox5a, cox5b, cox17 and hbg-3 expression and prolonged homing flight duration. Our results suggest that flight behavior can impact gene expression and exposure to pesticides adversely affects the expression of genes that are important for maintaining optimal flight capacity. Our laboratory-based experiment showed significantly altered expression levels of cox5a, cox6c, and cox17. However, further work is needed to identify transcriptional profiles responsible for prolonged homing flight duration.

Pesticide residues are frequently detected in surface waters, with several compounds known to adversely affect aquatic organisms. Gammarids are particularly suitable indicator organisms for assessing the sublethal effects of such contaminants due to their high sensitivity and their central ecological role in freshwater ecosystems. While behavioral endpoints and feeding rates have been commonly used to evaluate sublethal pesticide effects, gene expression changes have received comparatively little attention, despite their proven value in other ecotoxicological contexts. This study investigates the potential of gene expression as a sensitive sublethal endpoint in gammarids collected from natural populations. A laboratory exposure experiment was conducted using the model pesticides azoxystrobin and acetamiprid, both of which are regularly detected in surface waters. Gammarids collected from the wild were exposed under controlled conditions to sublethal concentrations of the test substances. Subsequently, RNA sequencing (RNA-seq) was performed to characterize genome-wide transcriptional responses. Two independent exposure and sequencing experiments were carried out, resulting in the identification of 145 and 326 differentially expressed transcripts per experiment when comparing exposed animals to controls. Gene ontology (GO) term enrichment analyses revealed significant effects on metabolic processes, cell proliferation, and cell differentiation. Notably, the two experimental runs yielded distinct transcriptional profiles, with minimal overlap in differentially expressed transcripts despite the use of gammarids from the same population and the short interval (12 days) between experiments. The study demonstrates the applicability of transcriptomic analyses for detecting sublethal pesticide effects in field-collected gammarids and provides a practical workflow for the evaluation of RNA-seq data in non-model organisms. At the same time, it highlights important limitations, including high genetic variability within wild populations and incomplete transcriptome annotation, which together contribute to inconsistencies across repeated experiments.