Abstract
Fractionation of H. haemachatus, N. nigricollis, N. nivea and N. melanoleuca venoms using Amberlite CG-50 and (NH4)HCO3 elution gradient chromatography yielded 11-13 fractions for each venom. One fraction, F X, from H. haemachatus, two fractions, F X and F XI, from N. nigricollis and one fraction, F VIII, from N. melanoleuca venoms possessed the whole of ocular activity of the venoms. The fractions were the only venom fractions that caused cardiac depressant activity; their effect was reversed by raising Ca++ concentration in the physiological solution; they did not influence the twitches of the phrenic nerve hemidiaphragm and guinea-pig ileum preparations. Further purification of the fractions on Sephadex G-50 followed by fractionation on Amberlite CG-50 yielded fractions free from phospholipase A2 activity but possessing the same ocular effects. Similarly, the cardiotoxin from commercial N. nigricollis venom caused the same ocular effects as the crude venom and its purified cardiotoxic fractions. All cardiotoxic fractions as well as N. nigricollis cardiotoxin, caused extensive chemosis, blepharitis and corneal opacification with corneal and subconjunctival neovascularization. On a weight basis, the cardiotoxins were weaker in their oculotoxic activity than the corresponding parent crude venoms possibly because of the potentiating effect of phospholipase A2 in the crude venoms. It is postulated that in spitting cobras the cardiotoxins are responsible for the corneal opacification syndrome. In other cobra venoms the stable binding of cardiotoxins with acidic proteins limits their possible ocular effects. Only in the venoms of the spitting species are the cardiotoxins present in an appropriately free form to cause the ocular opacification syndrome.