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Characterization of Indian Common Krait (Bungarus Caeruleus) Venom

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Characterization of Indian Common Krait (Bungarus Caeruleus) Venom ISSN 2320-5407 International Journal of Advanced Research (2014) Journal homepage: http://www.journalijar.com INTERNATIONAL JOURNAL OF ADVANCED RESEARCH CHARACTERIZATION OF INDIAN COMMON KRAIT (BUNGARUS CAERULEUS) VENOM Dissertation submitted to H. P. UNIVERSITY In partial fulfillment of the requirements For the award of the degree of M.Sc. MICROBIOLOGY ΙV SEMESTER DEGREE EXAMINATION MAY, 1988 ANJU DHIR CENTRAL RESEARCH INSTITUTE KASAULI HIMACHAL PRADESH DISTRICT INDIA 1 ISSN 2320-5407 International Journal of Advanced Research (2014) CONTENTS SUBJECT PAGE Acknowledgement 3 List of Tables 4 List of Figures 5 Introduction 6 Aim 7 Review of Literature 7 Materials and Methods 11 Results 13 Discussion 29 References 32 2 ISSN 2320-5407 International Journal of Advanced Research (2014) Acknowledgement I would like to express my deepest appreciation and gratitude to Late Dr. B. Das, Head of Antisera Department of Central Research Institute, Kasauli, Himachal Pradesh, India for his keen supervision and continuous encouragement. He offered deep experience, unlimited support, valuable suggestions and sincere advice. It was a great honor to work under his supervision. To him, I owe a great deal of gratitude and thanks. Anju Dhir 3 ISSN 2320-5407 International Journal of Advanced Research (2014) List of Tables Table Title Page Number 1. Determination of LD50 of Indian common krait (B. caeruleus) venom 13 by subcutaneous route 2. Determination of LD50 of Indian common krait (B. caeruleus) venom 14 by intramuscular route 3. Determination of LD50 of Indian common krait (B. caeruleus) venom 14 by intraperitoneal route 4. Determination of LD50 of Indian common krait (B. caeruleus) venom 15 by intravenous route 5. Determination of LD50 of Indian common krait (B. caeruleus) venom 16 by intracerebral route 6 Toxicity ofIndian common krait (B. caeruleus) venom by different 17 routes of inoculations 7. Statistical analysis of the results of LD50of Indian common krait (B. 18 caeruleus) venom 8. Hemolytic activity of Indian common krait (B. caeruleus) venom 1% 19 mice Red blood cells 9. Hemolytic activity of Indian common krait (B. caeruleus) venom 1% 20 rat Red blood cells 10. Hemolytic activity of Indian common krait (B. caeruleus) venom 1% 21 guinea pig Red blood cells 11. Hemolytic activity of Indian common krait (B. caeruleus) venom 1% 22 rabbit Red blood cells 12. Hemolytic activity of Indian common krait (B. caeruleus) venom 1% 23 sheep Red blood cells 13. Hemolytic activity of Indian common krait (B. caeruleus) venom 1% 24 fowl Red blood cells 14. Hemolytic activity of Indian common krait (B. caeruleus) venom 1% 25 mule Red blood cells 15. Hemolytic activity of Indian common krait (B. caeruleus) venom 1% 26 horse Red blood cells 16. Hemolytic activity of Indian common krait (B. caeruleus) venom 1% 27 human Red blood cells 17. Hemolytic activity of Indian common krait (B. caeruleus) venom 28 using erythrocytes from different species of animals 18. Statistical analysis of the results of HD50of Indian common krait (B. 28 caeruleus) venom 4 ISSN 2320-5407 International Journal of Advanced Research (2014) List of Figures Table Title Page Number 1. Bar diagram of Probit mortality of Indian common krait (B. caeruleus) 13 venom by subcutaneous route 2. Bar diagram of Probit mortality of Indian common krait (B. caeruleus) 14 venom by intramuscular route 3. Bar diagram of Probit mortality of Indian common krait (B. caeruleus) 15 venom by intraperitoneal route 4. Bar diagram of Probit mortality of Indian common krait (B. caeruleus) 15 venom by intravenous route 5. Bar diagram of Probit mortality of Indian common krait (B. caeruleus) 16 venom by intracerebral route 6.. Comparative results of various routes of inoculations 17 7. Hemolysis of 1% mice erythrocytes by Indian common krait (B. 19 caeruleus) venom 8. Hemolysis of 1% rat erythrocytes by Indian common krait (B. 20 caeruleus) venom 9. Hemolysis of 1% guinea pig erythrocytes by Indian common krait (B. 21 caeruleus) venom 10. Hemolysis of 1% rabbit erythrocytes by Indian common krait (B. 22 caeruleus) venom 11. Hemolysis of 1% sheep erythrocytes by Indian common krait (B. 23 caeruleus) venom 12. Hemolysis of 1% fowl erythrocytes by Indian common krait (B. 24 caeruleus) venom 13. Hemolysis of 1% mule erythrocytes by Indian common krait (B. 25 caeruleus) venom 14. Hemolysis of 1% horse erythrocytes by Indian common krait (B. 26 caeruleus) venom 15. Hemolysis of 1% human erythrocytes by Indian common krait (B. 27 caeruleus) venom 5 ISSN 2320-5407 International Journal of Advanced Research (2014) INTRODUCTION Snakes are the limbless vertebrates and belong to the class of living creatures known as reptiles which include all the living orders of crocodiles, turtles, tortoises and lizards.These reptiles came into existence long before the mammals and birds were born on this earth. Fossil remains of snakes have been found in the later cretaceous and early tertiary periods in the world’s history. Herpetologists agree that the snakes and lizards share a common ancestor having labial glands on both jaws. Only fossil record available in this direction has been found in case of the marine snake like lizards. Underwood, 1969 also has the opinion that the origin of snakes precedes the modern lizards. The snakes have originated from lizards or some prelizard like ancestors. Snakes are scattered all over the world with the exception of Newzeland where they have never known to exist. Snakes like heat, it means life and vitality to them, and therefore the snakes are most abundant in the tropical and subtropical regions of the globe, in forests and places with thick vegetation. This type of habitat is favored by snakes as it gives necessary warmth to quicken their sluggish vital forces. When the temperature of the air is fairly warm snakes may be seen at any hour of the day or night. The venomous snakes are classified according to their morphological characteristics and comprise five families: crotalidae e.g. pit vipers; viperidae e.g. vipers; elapidae e.g. cobras and kraits; hydrophidae e.g. sea snakes and colubridae e.g. colubrids. The venomous snakes have a pair of venom glands which are lying on either side of the skull beneath the skin, situated just under and behind the eyes. The venom glands may be true salivary glands with or without digestive enzymes. Without the venom it may be very difficult or even impossible for snakes to obtain their food. About 2000 different types of snakes exist and nearly 400 are known to be venomous. Nearly 20,000 people die every year from snake bite in India (Ahuja & Singh, 1954). Dr. K.C.Chughfrom P.G.I. in 1988 has also reported that the global annual mortality rate from snake bite is about 40,000; out of this nearly 10,000 deaths occur in India, which is highest as compared to any other country in the world. The common four speciesresponsible for these casualties belong to two families: viperidae and elapidae. The venom of elapids is neurotoxic and death occurs due to respiratory failure. The Indian common krait and cobra belong to this family. Whereas the venom of vipers cause hemorrhagic effect and death occurs due to cardiac failure – Russel’s viper and Echis carinatus belong to this family. The snakes are frequently found in and around the human dwellings. In autumn they find some cozy place in the thatch, under the roof or floor, down a hole, in a stack of timber, dung heap or in fodder, in order to hibernate in winter season. Moreover, the snakes are highly evolved creatures and do not depend much on oxygen like most of the other animals. Due to this quality and by virtue of their specifically modified organs they are able to penetrate into dense overgrown vegetation of the tropics where the smaller reptiles are found in less number. The snakes may take a heavy toll of life every year but in nature they keep the growth of fast breeding creatures in check. In hot countries insects and rodents are a nuisance to the farmers as they damage the crops. Owing to the long tapering bodies, the snakes are capable to penetrate into the innermost dwellings of these animals to feed upon them. Thus they act as the part of nature’s agent in checking their rapid increase. The snakes are also a valuable source of venom which is used in various research fields as the snake venoms retain many of their biological activities over long periods of time. It was not until the 17th century that systematic studies of the venoms were made in a manner that may be regarded as positive. In 1664, Francesco Redi demonstrated for the first time that the snake venom must be injected under the skin to produce their characteristic effects as certain venoms were found to be harmless when taken by mouth. Systematic studies on the biological action of 6 ISSN 2320-5407 International Journal of Advanced Research (2014) snake venoms were undertaken by Flexner in 1863-1954at the Rockfeller institute in New York. The reports from these studies described the effects of various snake venoms in producing hemolysis and other toxic effects. But the chemists were among the first to consider the venom with interest, and they defined their composition. They saw the effect of common reagents on snake venom and tried to extract the toxic constituents. They observed that certain reagents destroyed the venom while others caused precipitation of the toxic components. In 1938, a crystallisable protein ‘crotoxin’ was separated from venom of Crotalus durissus terrificus (Slotta and Frankel Conrat, 1938). After this work many reports about procedures for purifying snake venom enzyme appeared. The older methods for isolation and purification of the venom components include heat and pH treatment, alcohol fractionation, free boundary electrophoresis and ultracentrifugation.
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