De Novo Transcriptome Analysis of the Venom of Latrodectus geometricus with the Discovery of an Insect-Selective Na Channel Modulator

The brown widow spider, Latrodectus geometricus, is a predator of a variety of agricultural insects and is also hazardous for humans. Its venom is a true pharmacopeia representing neurotoxic peptides targeting the ion channels and/or receptors of both vertebrates and invertebrates. The lack of transcriptomic information, however, limits our knowledge of the diversity of components present in its venom. The purpose of this study was two-fold: (1) carry out a transcriptomic analysis of the venom, and (2) investigate the bioactivity of the venom using an electrophysiological bioassay. From 32,505 assembled transcripts, 8 toxin families were classified, and the ankyrin repeats (ANK), agatoxin, centipede toxin, ctenitoxin, lycotoxin, scorpion toxin-like, and SCP families were reported in the L. geometricus venom gland. The diversity of L. geometricus venom was also uncovered by the transcriptomics approach with the presence of defensins, chitinases, translationally controlled tumor proteins (TCTPs), leucine-rich proteins, serine proteases, and other important venom components. The venom was also chromatographically purified, and the activity contained in the fractions was investigated using an electrophysiological bioassay with the use of a voltage clamp on ion channels in order to find if the neurotoxic effects of the spider venom could be linked to a particular molecular target. The findings show that U24-ctenitoxin-Pn1a involves the inhibition of the insect sodium (Nav) channels, BgNav and DmNav. This study provides an overview of the molecular diversity of L. geometricus venom, which can be used as a reference for the venom of other spider species. The venom composition profile also increases our knowledge for the development of novel insecticides targeting voltage-gated sodium channels.

Eggshell spheres protect brown widow spider ( Latrodectus geometricus ) eggs from bacterial infection

Eggs provide a rich source of nutrients for the developing embryo, making them a favoured food source for other organisms as well. Several defence mechanisms have evolved to protect the developing embryos against microbial threats. In this article, we elucidate the defence strategy of brown widow spider ( Latrodectus geometricus ) eggs against bacteria. Antibacterial activity was shown by inhibition of bacterial growth on agar plate, liquid culture and retarded biofilm formation. The defence strategy against bacterial invasion was demonstrated in the whole egg, whole egg extract, egg surface extract, eggshell and eggshell extract. The source and characteristics of this antibacterial activity are distinctive and stem in part from a dense layer of spheres covering the egg surface, likely originated from the oviposition fluid. The spheres are rich in low-molecular-weight proteins, yet their exact composition remains unknown. In this study, we demonstrate that the egg surface is hydrophobic, while the spheres are superhydrophilic. Egg surface roughness and hydrophobicity combined with its antibacterial chemical properties reduce the ability of bacteria to grow on the egg surface. Understanding the properties of these unique structures may contribute significantly to our knowledge of how nature deals with bacterial infections.