Vol. 9, Special Issue 12, Part S (2025)

The present study was undertaken to analyze the diversity and composition of spider venom in selected species from Chhattisgarh, India, with emphasis on biochemical complexity, enzymatic activity and ecological relevance. Six spider species belonging to five families-Hippasa agelenoides (Lycosidae), Pholcus phalangioides (Pholcidae), Cyrtophora citriola and Argiope aemula (Araneidae), Oxyopes salticus (Oxyopidae) and Heteropoda venatoria (Sparassidae)-were selected based on their abundance, ecological guild and habitat diversity. Venom was extracted using mild electrical stimulation and analyzed for yield, protein concentration and molecular composition. Venom yield ranged from 1.8 ± 0.3 µL in Pholcus phalangioides to 6.4 ± 0.8 µL in Heteropoda venatoria. Bradford assay revealed significant interspecific variation in protein concentration, with the highest levels recorded in Argiope aemula (2.63 ± 0.18 µg/µL) and Cyrtophora citriola (2.21 ± 0.16 µg/µL). SDS-PAGE and RP-HPLC analyses demonstrated species-specific protein profiles and varying degrees of venom complexity, with orb-weaving spiders and large wandering hunters exhibiting greater protein diversity and enzymatic activity. Enzymatic assays indicated the presence of phospholipase A₂, proteases and antimicrobial components, particularly in Argiope aemula, Cyrtophora citriola and Heteropoda venatoria. Statistical analysis confirmed significant differences in venom protein concentration among species (p < 0.05). The study reveals a strong association between venom composition, ecological guild and predatory strategy, highlighting orb-weaving spiders as reservoirs of highly complex venoms. These findings provide baseline information on spider venom diversity in Chhattisgarh and underscore the potential of regional spider venoms for future pharmacological and bioinsecticidal research.