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Honeybees secrete 2-heptanone (2-H) from their mandibular glands when they bite. Researchers have identified several possible functions: 2-H could act as an alarm pheromone to recruit guards and soldiers, it could act as a chemical marker, or it could have some other function. The actual role of 2-H in honeybee behaviour remains unresolved. In this study, we show that 2-H acts as an anaesthetic in small arthropods, such as wax moth larva (WML) and Varroa mites, which are paralysed after a honeybee bite. We demonstrated that honeybee mandibles can penetrate the cuticle of WML, introducing less than one nanolitre of 2-H into the WML open circulatory system and causing instantaneous anaesthetization that lasts for a few minutes. The first indication that 2-H acts as a local anaesthetic was that its effect on larval response, inhibition and recovery is very similar to that of lidocaine. We compared the inhibitory effects of 2-H and lidocaine on voltage-gated sodium channels. Although both compounds blocked the hNav1.6 and hNav1.2 channels, lidocaine was slightly more effective, 2.82 times, on hNav.6. In contrast, when the two compounds were tested using an ex vivo preparation–the isolated rat sciatic nerve–the function of the two compounds was so similar that we were able to definitively classify 2-H as a local anaesthetic. Using the same method, we showed that 2-H has the fastest inhibitory effect of all alkyl-ketones tested, including the isomers 3- and 4-heptanone. This suggests that natural selection may have favoured 2-H over other, similar compounds because of the associated fitness advantages it confers. Our results reveal a previously unknown role of 2-H in honeybee defensive behaviour and due to its minor neurotoxicity show potential for developing a new local anaesthetic from a natural product, which could be used in human and veterinary medicine.

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Background With numerous endemic subspecies representing four of its five evolutionary lineages, Europe holds a large fraction of Apis mellifera genetic diversity. This diversity and the natural distribution range have been altered by anthropogenic factors. The conservation of this natural heritage relies on the availability of accurate tools for subspecies diagnosis. Based on pool-sequence data from 2145 worker bees representing 22 populations sampled across Europe, we employed two highly discriminative approaches (PCA and FST) to select the most informative SNPs for ancestry inference. Results Using a supervised machine learning (ML) approach and a set of 3896 genotyped individuals, we could show that the 4094 selected single nucleotide polymorphisms (SNPs) provide an accurate prediction of ancestry inference in European honey bees. The best ML model was Linear Support Vector Classifier (Linear SVC) which correctly assigned most individuals to one of the 14 subspecies or different genetic origins with a mean accuracy of 96.2% ± 0.8 SD. A total of 3.8% of test individuals were misclassified, most probably due to limited differentiation between the subspecies caused by close geographical proximity, or human interference of genetic integrity of reference subspecies, or a combination thereof. Conclusions The diagnostic tool presented here will contribute to a sustainable conservation and support breeding activities in order to preserve the genetic heritage of European honey bees.

-Honeybees face several predators and their ability to express collective defence behaviour is one of their major life traits that promote colony survival. We discovered that, while confronting attacks by the Oriental hornet Vespa orientalis, Apis mellifera cypria honeybees engage in a distinct acoustic behaviour: they produce a characteristic hissing sound of unexpectedly high frequency. When recording and analysing these hissing sounds during an extended sample of artificial attacks by hornets, we found that honeybees can produce sounds covering a wide frequency spectrum with a dominant frequency around 6 kHz. Notably, these acoustic emissions are distinct from the background noise of neighbouring flying bees. These results provide a detailed description of the sounds generated by A. m. cypria when defending their nest against hornets, and they could be used for future research to better understand the biological function of the acoustic behaviour in honeybees' colony defence.hissing sound / colony defence / Apis mellifera cypria / Vespa orientalis / prey-predator contest

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Guards of Cyprian honey bee colonies, Apis mellifera cypria, display a great defensive behaviour against hornets' attacks. The deformed wing virus (DWV) and the kakugo virus (KV) genomes are very similar, but unlike KV, the presence of DWV is not related to honey bees' aggressiveness. This discrepancy is further discussed.

Honey is a natural food with a long history as a traditional medicine because of its many biological characteristics, including antimicrobial, antioxidant, anti-tumor and anti-inflammatory properties. In this study, the antimicrobial actions of eight different honeys from Lemnos island (north-eastern Greece) plus manuka honey (from New Zealand, UMF 30+, licensed in many countries as topical medical preparation) were evaluated against 10 clinically relevant bacteria, including five Gram-positive and five Gram-negative. To achieve this, an agar well diffusion assay measured the diameter of inhibition zones (mm) of two selected concentrations for each honey (25% and 12.5% v/v). The minimum inhibitory and bactericidal concentrations (MIC and MBC) of each sample were also calculated and compared against two representative bacterial species (Salmonella Typhimurium and Staphylococcus aureus) using broth microdilution and agar spot methods, respectively. The pH, water activity (aw), 5-hydroxymethylfurfural (HMF) and diastase levels, together with the pollen type and content of each honey, were also determined. Results revealed that all the Lemnos honeys presented antibacterial action, which for some samples was like that of manuka. These all had an acidic pH (3.61 ± 0.04), with a aw ≤ 0.60, while it is worth noting that those found to display the strongest antibacterial actions also presented the lowest HMF content, together with the highest diastase values, both of the latter being used as quality parameters. Pollen composition of the Lemnos honeys was multifloral, underlining the rich plant biodiversity encountered on the island. To summarize, Lemnos honeys could be further exploited as natural antimicrobial systems for use in foods and medicine.