Objective  To investigate the components of snake venom and the dynamic changes in protein expression in the exudates of necrotic tissues affected by the Naja atra venom in Guangxi Bama miniature pigs.

Methods  Guangxi Bama miniature pigs were injected with Naja atra venom to establish a local tissue necrosis model due to snakebite poisoning. Exudates from the necrotic tissues of the pigs were collected at 6 h, 12 h, 24 h, 36 h, and 48 h post-injection. Label-free proteomics techniques were employed to identify and analyze the composition and dynamic changes of snake venom proteins in the exudates. KEGG and GO pathway enrichment analyses were performed on the identified proteins to gain deeper insights into the biological functions related to snake venom proteins.

Results  The biological behavior of the Guangxi Bama miniature pigs post-snakebite, pathological results of local muscle tissues, and changes at the injection site were used to evaluate the animal model. The pigs exhibited symptoms such as rapid respiration, muscle tissue necrosis, and local wound ulceration, consistent with clinical characteristics, indicating the successful establishment of a Guangxi Bama miniature pig model of local tissue necrosis induced by Naja atra venom. A total of 40 snake venom proteins were identified in the exudates of the local tissue necrosis in the Guangxi Bama miniature pigs, including three-finger toxins, phospholpase A2, cysteine- rich secretory proteins, snake venom metalloproteinases, and nucleotidyl pyrophosphatase/phosphodiesterases, showing dynamic quantitative changes.

Conclusion  Various snake venom proteins, including three-finger toxins and phospholipase A2, are identified in the exudates of the tissues at different time points during the poisoning process of Guangxi Bama miniature pigs by Naja atra venom, suggesting their important roles in local tissue necrosis.