Exploring the ex vivo effects of Naja mossambica venom on the ultrastructure and viscoelastic properties of human blood.

Background: Venomous snake bites have been listed as a neglected tropical disease by the World Health Organization. The Mozambique spitting cobra (Naja mossambica) is found in Sub-Saharan African countries, and its venom has been identified to predominantly result in cytotoxic effects. However, there is limited evidence on the possible hemotoxic effects of this venom on human blood. Objectives: In this cross-sectional study, we investigated how Mozambique spitting cobra venom affects blood clot formation. Methods: Cell morphology and clot architecture were studied by using microscopy techniques. We also studied the effects of the venom on platelets by measuring platelet activity with the global thrombosis test, followed by analyzing the viscoelasticity with thromboelastography using a 0.025 ng/µL venom concentration. Results: The most prominent findings indicated that the viscoelastic profile in the venom-treated blood samples formed an unstable and elastic clot. The clot architecture seen with the scanning electron microscopy analysis showed an altered fibrin network and red blood cells, confirmed by the increased axial ratios, and aggregated platelets with spreading. Conclusion: These findings may offer insights into the species-specific effects of snake venom on human blood and add value to the clinical workup in confirming envenomation. Further research is needed to correlate the 20 minute whole blood clotting test with measurable values from the thromboelastography within the context of snake envenomation. This may offer a bridge between cost, early diagnosis, and treatment of snake envenomation in resource-constrained countries.

How the Cobra Got Its Flesh-Eating Venom: Cytotoxicity as a Defensive Innovation and Its Co-Evolution with Hooding, Aposematic Marking, and Spitting.

The cytotoxicity of the venom of 25 species of Old World elapid snake was tested and compared with the morphological and behavioural adaptations of hooding and spitting. We determined that, contrary to previous assumptions, the venoms of spitting species are not consistently more cytotoxic than those of closely related non-spitting species. While this correlation between spitting and non-spitting was found among African cobras, it was not present among Asian cobras. On the other hand, a consistent positive correlation was observed between cytotoxicity and utilisation of the defensive hooding display that cobras are famous for. Hooding and spitting are widely regarded as defensive adaptations, but it has hitherto been uncertain whether cytotoxicity serves a defensive purpose or is somehow useful in prey subjugation. The results of this study suggest that cytotoxicity evolved primarily as a defensive innovation and that it has co-evolved twice alongside hooding behavior: once in the Hemachatus + Naja and again independently in the king cobras (Ophiophagus). There was a significant increase of cytotoxicity in the Asian Naja linked to the evolution of bold aposematic hood markings, reinforcing the link between hooding and the evolution of defensive cytotoxic venoms. In parallel, lineages with increased cytotoxicity but lacking bold hood patterns evolved aposematic markers in the form of high contrast body banding. The results also indicate that, secondary to the evolution of venom rich in cytotoxins, spitting has evolved three times independently: once within the African Naja, once within the Asian Naja, and once in the Hemachatus genus. The evolution of cytotoxic venom thus appears to facilitate the evolution of defensive spitting behaviour. In contrast, a secondary loss of cytotoxicity and reduction of the hood occurred in the water cobra Naja annulata, which possesses streamlined neurotoxic venom similar to that of other aquatic elapid snakes (e.g., hydrophiine sea snakes). The results of this study make an important contribution to our growing understanding of the selection pressures shaping the evolution of snake venom and its constituent toxins. The data also aid in elucidating the relationship between these selection pressures and the medical impact of human snakebite in the developing world, as cytotoxic cobras cause considerable morbidity including loss-of-function injuries that result in economic and social burdens in the tropics of Asia and sub-Saharan Africa.

The effect of physiological levels of South African puff adder (Bitis arietans) snake venom on blood cells: an in vitro model.

A significant burden of illness is caused globally by snakebites particularly by the puff adder, Bitis arietans. Presently there is no reliable and rapid method to confirm envenomation on blood chemistry; although coagulation parameters like prothrombin time, partial thromboplastin time, international normalized ratio and also serum electrolytes are tested. Here, we found that direct in vitro exposure of physiological relevant whole venom levels to human healthy blood (N = 32), caused significant physiological changes to platelet activity using a hematology analyzer, and measuring occlusion time, as well as lyses time, with the global thrombosis test (GTT). Disintegrated platelets were confirmed by scanning electron microscopy (SEM). We also confirmed the pathologic effects on erythrocytes (RBCs) (visible as eryptotic RBCs), by looking at both light microscopy and SEM. Thromboelastography showed that no clot formation in whole blood could be induced after addition of whole venom. We propose further clinical studies to investigate the use of light microscopy smears and hematology analyzer results immediately after envenomation, as a possible first-stage of clinical confirmation of envenomation.