From birth to bite: the evolutionary ecology of India's medically most important snake venoms

Venoms of N. naja and D. russelii individuals of various developmental stages, including neonates (<30 days), juveniles (between 1 to 12 months), and mature individuals (>36 months), were sampled with permission from the state forest department of Karnataka. 226 individuals and 9 clutches were examined, including periodic venom collection (every three months) from the same N. naja and D. russelii juvenile individuals to track ontogenetic changes across time. Freshly collected venoms were flash-frozen in liquid nitrogen, lyophilised and stored at -80° C. Venoms were fractionated using RP-HPLC and SDS-PAGE and subjected to tandem mass spectrometry for toxin identification. The relative abundance of each toxin family were estimated and compared across different developmental stages.