Comparison of external characteristics and crude venom composition of five species of vermivorous cone snails. (a) External characteristics of common vermivorous cone snails found in the South China Sea. From left to right: C. generalis, C. caracteristicus, C. betulinus, C. quercinus, and C. tesselatus. (b) Schematic representation of venom tube dissection for the five species of cone snail. From top to bottom: venom glands and ducts of C. generalis, C. caracteristicus, C. betulinus, C. quercinus, and C. tesselatus. (c) SDS-PAGE analysis of crude venom from the five species of cone snail. M indicates the standard marker. Lanes 1–5 correspond to the crude venom of C. tesselatus, C. caracteristicus, C. quercinus, C. betulinus, and C. generalis, respectively, with a loading volume of 10 μL. (d) RP-HPLC analysis of crude venom from the five species of cone snail.

Cluster analysis of mitochondrial COI gene sequence and HPLC fingerprint of five species of vermivorous cone snails. (a) Based on COI gene sequences of five species of vermivorous cone snails, a phylogenetic tree was constructed using the ML method. (b) Cluster diagram based on RP-HPLC data of five species of crude venom from vermivorous cone snails.

Insecticidal effects and LD₅₀ values of crude venom from five species of vermivorous cone snail on Spodoptera litura larvae at various concentrations. (a–e) The insecticidal effects of crude venoms from C. quercinus, C. tesselatus, C. caracteristicus, C. betulinus, and C. generalis on Spodoptera litura at a specified concentration, n = 3. (f) The LD₅₀ values of these five species of vermivorous cone snail against Spodoptera litura larvae at varying concentrations. ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus blank control (BC) group.

Cytotoxicity assays of crude venom from five species of vermivorous cone snail on ovarian cancer cells at different concentrations. (a–e) Cytotoxicity assays of crude venom from C. quercinus, C. tesselatus, C. caracteristicus, C. betulinus, and C. generalis on ovarian cancer cells at predetermined concentrations, n = 3. (f) IC₅₀ value for the crude venom of four species of vermivorous cone snail on ovarian cancer cells at varying concentrations. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 compared to the blank control (BC) group.

Toxicity analysis of crude venom from five species of vermivorous cone snail on zebrafish at varying concentrations. (a–e) The mortality rates of zebrafish exposed to crude venom from C. tesselatus, C. caracteristicus, C. quercinus, C. betulinus, and C. generalis at specific concentrations, n = 3. (f) The LD₅₀ values for the crude venom of C. tesselatus and C. caracteristicus. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus the blank control (BC) group.

Analgesic effects of crude venoms from five species of vermivorous cone snail on zebrafish larvae. (a) Analgesic effects of crude venoms from five species of vermivorous cone snail on zebrafish larvae, evaluated based on their swimming distance, n = 12. (b) Motion trajectory diagrams for the blank control group, acetic acid model group, and positive drug (Tramadol and Lidocaine) group. Motion trajectories were recorded every 5 s, with instantaneous velocity indicated by color: black (<2 mm/s), green (2–8 mm/s), and red (>8 mm/s). (c) Motion trajectory diagram of zebrafish larvae treated with crude venom from C. caracteristicus. (d) Analgesic effects of C. caracteristicus crude venom on zebrafish larvae, n = 12. (e) Line graph showing movement changes in zebrafish larvae treated with C. caracteristicus venom, measured every two minutes. ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus the acetic acid group.