Quantitative Characterization of the Hemorrhagic, Necrotic, Coagulation-Altering Properties and Edema-Forming Effects of Zebra Snake (Naja nigricincta nigricincta) Venom

Abstract

This study was designed to investigate the cytotoxicity and haemotoxicity of the Western barred (zebra) spitting cobra (Naja nigricincta nigricincta) venom to help explain atypical and inconsistent reports on syndromes by Namibian physicians treating victims of human ophidian accidents. Freeze-dried venom milked from adult zebra snakes was dissolved in phosphate buffered saline (PBS) for use in this study. Haemorrhagic and necrotic activity of venom were studied in New Zealand albino rabbits. Oedema-forming activity was investigated in 10-day-old Cobb500 broiler chicks. Procoagulant and thrombolytic activity was investigated in adult Kalahari red goat blood in vitro. The rabbit skin minimum hemorrhagic dose (MHD) for N. n. nigricincta was 9.8μg. The minimum necrotizing dose (MND) for N. n. nigricincta venom was 12.2μg. The N. n. nigricincta venom showed linear dose-dependent procoagulant activity on goat blood (p<0.05). Likewise, N. n. nigricincta venom showed linear dose-dependent thrombolytic activity on goat blood (p<0.05, n = 6). Subplantar injection of N. n. nigricincta venom (25μg, 50μg, 75μg, and 100μg) into chick paw resulted in peak oedema of 35.5%, 38.5%, 42.9%, and 47.5%, respectively, two hours after injection. Paw oedema subsided within five hours to a mean volume ranging from 5% (25μg venom) to 17.6% (100μg venom). In conclusion, though N. n. nigricincta belongs to the genus Elapidae, the current study has shown its venom to possess potent hemorrhagic, necrotic (cytotoxic), and paradoxically, both procoagulant and thrombolytic activity. The authors propose further work to fractionate, isolate, and elucidate the structure of the various N. n. nigricincta venom toxins as a prelude to the development of an antivenom.

Figure 1

Haemorrhagic lesions on internal aspect of rabbit skin 24 hours after intradermal injection of varying doses of N. n. nigricincta venom.

Figure 2

The dose-dependent hemorrhagic activity of N. n. nigricincta venom on adult rabbit skin.

Figure 3

The dose-dependent necrotic activity of N. n. nigricincta venom on adult rabbit skin.

Figure 4

The dose-dependent anticoagulative activity of heparin on Kalahari red goat blood.

Figure 5

Dose-dependent coagulative (thrombotic) activity of N. n. nigricincta venom on Kalahari red goat blood.

Figure 6

Comparison of dose-dependent thrombolytic activity of heparin and N. n. nigricincta venom on Kalahari red goat blood clots.

Figure 7

Dose-dependent oedema-forming effect of N. n. nigricincta venom on 10-day-old chick paw.