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Australian funnel-web spiders are arguably the most venomous spiders in the world, with much research focusing on this aspect of their biology. However, other aspects related to their life history, ecology and behaviour have been overlooked. For the first time, we assessed repeatability, namely risk-taking behaviour, aggressiveness and activity in the contexts of predation, conspecific tolerance and exploration of a new territory in four species of Australian funnel-web spiders: two are closely related, Hadronyche valida and H. infensa , and two have overlapping distributions but occupy different habitats, H. cerberea and Atrax robustus. We also compared behaviors between species. At the species level, we found that H. valida showed consistency in risk-taking behavior when exposed to a predator stimulus, aggressiveness against conspecifics, and exploration of a new territory. In contrast, in the other species, only A. robustus showed repeatability in the context of exploration of a new territory. These results suggest that some behavioral traits are likely more flexible than others, and that the repeatability of behaviors may be species-specific in funnel-webs. When we compared species, we found differences in risk-taking behavior and defensiveness. This study provides novel insights to understanding variation in behavioral traits within and between species of funnel-web spiders, suggesting that some behavioral traits are likely context and/or species dependent, as a result of their evolutionary history. These findings provide key insights for understanding the ecological role of behavior and venom deployment in venomous animals, and a greater understanding of behavior in these medically significant and iconic spiders that are of conservation concern. [ABSTRACT FROM AUTHOR]

The stonefish Synanceia verrucosa and Synanceia horrida are arguably the most venomous fish species on earth and the culprits of severe stings in humans globally. Investigation into the venoms of these two species has mainly focused on protein composition, in an attempt to identify the most medically relevant proteins, such as the lethal verrucotoxin and stonustoxin components. This study, however, focused on medically relevant small molecules, and through nuclear magnetic resonance, mass spectroscopy, and fractionation techniques, we discovered and identified the presence of three molecules new to stonefish venom, namely γ-aminobutyric acid (GABA), choline and 0-acetylcholine, and provide the first report of GABA identified in a fish venom. Analysis of the crude venoms on human nicotinic acetylcholine receptors (nAChRs) and a GABA A receptor (GABA A R) showed S. horrida venom could activate neuronal (α7) and adult muscle-type (α1β1δε) nAChRs, while both crude S. horrida and S. verrucosa venoms activated the GABA A R (α1β2γ2). Cytotoxicity studies in immunologically relevant cells (human PBMCs) indicated the venoms possess cell-specific cytotoxicity and analysis of the venom fractions on Na + channel subtypes involved in pain showed no activity. This work highlights the need to further investigate the small molecules found in venoms to help understand the mechanistic pathways of clinical symptoms for improved treatment of sting victims, in addition to the discovery of potential drug leads., (© 2024 The Author(s). FEBS Open Bio published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.)