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. 2022 Mar 14:28:e20210040.
doi: 10.1590/1678-9199-JVATITD-2021-0040. eCollection 2022.

Compositional and toxicological investigation of pooled venom from farm-raised Naja atra

Gang Xiao  1   2 , Junqi Liu  1   3 , Lingfeng Peng  1   2 , Yang Yang  1 , Zhiliang Sun  1   2
Affiliations

Compositional and toxicological investigation of pooled venom from farm-raised Naja atra

Gang Xiao et al. J Venom Anim Toxins Incl Trop Dis. .

Abstract

Background: Naja atra is a venomous snake species medically relevant in China. In the current study, we evaluated the composition and toxicological profile of venom collected from farm-raised N. atra.

Methods: Venom was collected from third-generation captive bred N. atra on a snake farm in Hunan Province, China. The venom was analyzed using sodium dodecyl sulfate polyacrylamide gel electrophoresis and nano-liquid chromatography with electrospray ionization tandem mass spectrometry. In addition, hemolytic activity, median lethal dose, serum biochemical and histopathological parameters were accessed.

Results: N. atra venom proteome was dominated by phospholipase A2 (46.5%) and three-finger toxins (41.4 %), and a set of common low relative abundance proteins, including cysteine-rich secretory proteins (4.7%), NGF-beta (2.4%), snake venom metalloproteinase (1.5%), glutathione peroxidase (0.6%), vespryn (0.3%), and 5'-nucleotidases (0.2%) were also found. Furthermore, the venom exhibited direct hemolytic activity, neurotoxicity, myotoxicity, and high lethal potency in mice, with a subcutaneous median lethal dose of 1.02 mg/kg. Histopathological analysis and serum biochemical tests revealed that venom caused acute hepatic, pulmonary and renal injury in mice.

Conclusion: This study revealed the composition and toxicity of venom collected from farm-raised N. atra, thereby providing a reference for the analysis of venom samples collected from captive-born venomous snakes in the future.

Keywords: Myotoxicity; Naja atra; Neurotoxicity; Phospholipase A2; Three-finger toxins.

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Conflict of interest statement

Competing interests: The authors declare that they have no competing interests.

Figures

Figure 1.
Figure 1.. SDS-PAGE and proteomes of N. atra venom. (A) Protein separation of N. atra venom (20 μg) on 15% SDS-PAGE under reducing conditions. (B) Composition of N. atra venom according to protein families, expressed as percentages of the total protein content. N. atra: Naja atra; PLA 2: phospholipase A2; 3FTx: three-finger toxin; CRISP: cysteine-rich secretory protein; NGF-beta: venom nerve growth factor; SVMP: snake venom metalloproteinase; PDE: phosphodiesterase; 5ʹNUC: 5ʹ-nucleotidases; GLU: glutathione peroxidase; UP: uncharacterized protein; SNTX: short-chain α-neurotoxin; LNTX: long-chain α-neurotoxin; WNTX: weak neurotoxin.
Figure 2.
Figure 2.. Direct hemolysis caused by N. atra venom in rabbit erythrocytes. N: negative control (saline); P: positive control (distilled water). Data are presented as mean ± SEM (n = 6 each).
Figure 3.
Figure 3.. Histological analysis of hepatic, renal and pulmonary tissues of mice 6h, 12h and 24h after the injection of one LD50 (1.02 mg/kg, s.c.) of N. atra venom. The black arrows indicate the lesion area (scale bar: 50 μm).
Figure 4.
Figure 4.. Increases in the serum (A) ALT, (B) AST, (C) UN, (D) SCr and (E) CK levels of mice 1h after the injection of one LD50 (1.02 mg/kg, s.c.) of N. atra venom. The columns represent the mean +/- SEM (n = 6 each). **p < 0.01 and **p < 0.001.
See this image and copyright information in PMC

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