Naja siamensis, a cryptic species of venomous snake revealed by mtDNA sequencing

Because of possible variation in venom composition, an understanding of venomous snake systematics is of great importance for the optimization of antivenom treatment of snakebite patients. Intraspecific variation in the morphology of many venomous snakes complicates the definition and indentification of some species when allopatric populations are involved. Selectively neutral or near-neutral mtDNA sequences can reveal evolutionary relationships obscured by ecogenetically-caused morphological variation. We use comparative sequencing of the cytochrome oxidase subunit 1 gene to reveal the existence of a widespread, cryptic species of spiting cobra from southeast Asia. This species,Naja siamensis, is widely sympatric with other Asiatic cobra species. This may be of considerable medical significance, and calls for further research into venom composition in Asiatic cobras.     

Asiatic cobras: Systematics and snakebite

Summary The population affinities of the Asiatic cobras of the genusNaja are investigated, using multivariate analysis of a range of morphological characters. This complex, which was formerly thought to be monospecific, consists of at least eight full species. In some cases, species whose bites require different antivenoms occur sympatrically. The new understanding of the systematics of the Asiatic cobra complex calls for a reappraisal of cobra antivenom use in Asia, and for more research into venom composition.     

Asiatic cobras: systematics and snakebite

The population affinities of the Asiatic cobras of the genus Naja are investigated, using multivariate analysis of a range of morphological characters. This complex, which was formerly thought to be monospecific, consists of at least eight full species. In some cases, species whose bites require different antivenoms occur sympatrically. The new understanding of the systematics of the Asiatic cobra complex calls for a reappraisal of cobra antivenom use in Asia, and for more research into venom composition.     

Molecular basis of cardiotoxicity upon cobra envenomation

Various clinical manifestations leading to death have been documented in most cases of bites caused by venomous snakes. Cobra envenomation is an extremely variable process and known to cause profound neurological abnormalities. The complexity of cobra venom can induce multiple-organ failure, leading to death in case of severe envenomation. Intramuscular administration of Malayan spitting cobra (Naja sputatrix) crude venom at 1 g/g dose caused death in mice in approximately 3 h. Analysis of gene expression profiles in the heart, brain, kidney, liver and lung revealed 203 genes whose expression was altered by at least 3-fold in response to venom treatment. Of these, 50% were differentially expressed in the heart and included genes involved in inflammation, apoptosis, ion transport and energy metabolism. Electrocardiogram recordings and serum troponin T measurements indicated declining cardiac function and myocardial damage. This not only sheds light on the cardiotoxicity of cobra venom but also reveals the molecular networks affected during envenomation.Received 7 August 2004; received after revision 11 October 2004; accepted 4 November 2004     

Antimicrobial and cytolytic peptides of venomous arthropods

As a response to invading microorganisms, the innate immune system of arthropods has evolved a complex arrangement of constitutive and inducible antimicrobial peptides that immediately destroy a large variety of pathogens. At the same time, venomous arthropods have developed an additional offensive system in their venom glands to subdue their prey items. In this complex venom system, several enzymes, low-molecular-mass compounds, neurotoxins, antimicrobial and cytolytic peptides interact together, resulting in extremely rapid immobilization and/or killing of prey or aggressors. This review provides an overview of antimicrobial peptides identified in the hemolymph of venomous arthropods, and especially of cytolytic peptides in their venom. For these peptides a dual role is proposed: acting as antimicrobials as well as increasing the potency of the venom by influencing excitable cells.Received 17 March 2003; received after revision 11 June 2003; accepted 17 June 2003     

The effect of cobra venom factor (CVF) activation of the complement cascade on leukocyte circadian variation in the rat

Summary Administration of cobra venom factor (CVF) at different time periods over a 24-h-period produced a leukocytic response which varied according to the time of day the factor was given. The resulting leukocytic circadian rhythm was achieved by a marked variation and increase in the neutrophil population.This study was supported by U. S. P. H. S. Grant No. HL 16769 and Grant No. AI 09169.     

An alternative decomplementation method for the measurement of the survival of human erythrocytes transfused into rats

Summary An alternative technique for measuring the survival of⁵¹Cr-labelled human erythrocytes transfused into rats is described, in which aggregated human gamma-globulin is substituted for cobra venom factor as a decomplementing agent.Acknowledgments. This work was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo and Conselho Nacional de Desenvolvimento Cientifico e Tecnológico.     

Increased susceptibility ofTrypanosoma lewisi infected,or decomplemented rats toSalmonella typhimurium

Summary Rats infected withTrypanosoma lewisi or decomplemented by injection of cobra venom factor or complement activating factor of trypanosomes were found to be more susceptible to infection withSalmonella typhimurium. Decomplemented rats subsequently infected withT. lewisi developed higher blood parasitemia than did normalT. lewisi infected rats.This project is supported by the National Research Council of Canada grant A 0068 and a grant from the International Development Research Center.     

Myotoxic phospholipases A from snake venom,Pseudechis colletti,producing myoglobinuria in mice

Summary 2 proteins producing myoglobinuria in mice were isolated from the venom of the Australian elapid snakePseudechis colletti and identified as phospholipases A showing close similarities in amino acid composition to a similarly acting enzyme from a sea snake venom (Enhydrina schistosa).     

Chemistry of some potent animal toxins

Conclusion I have discussed in this article only the most active toxins, with the result that many interesting substances have been omitted, e.g. the toxins from bee and wasp venoms (apamin, melittin, etc.), of many amphibians (bufotoxins, etc.), ciguatoxins, and many more. Poisons are found in every phylum except birds. Shrews exemplify venomous mammals. One gets a good illustration of the number of poisonous animals by studying the monumental and impressive work byHalstead ¹⁰⁶ which consequently excludes terrestrial animals. An interesting fact in this connection is that there are about 20,000 species of spiders, most of which are poisonous.A toxin ranking list has to be included in an article of this kind. The list is, of course, far from complete. Data on molecular weights, mouse lethal doses, etc. are lacking for many potent toxins, such as the dysentery toxin, a neurotoxin with a toxicity comparable to that of the botulinus toxins¹⁰⁷, the toxins from the jelly fishChironex fleckeri ¹⁰⁶.A comparison on molar basis gives a better notion of the toxicities. Curare has about 1/30 of the toxicity of the curarimimetic snake venom neurotoxins, clearly indicating that curare has a much lower affinity for the acetylcholine receptor.Toxic organisms have developed during millions of years more and more refined toxins, and this evolution has probably brought into existence toxins against every physiological function. Neurochemistry is to a great extent unexplored. Progress in this field will in the nearest future depend on specific toxins from various natural sources. Toxins from spiders, scorpions, snakes, frogs, and fishes are therefore not mere curiosities but valuable tools for research on the molecular mechanisms of neural function and synaptic transmission.     

Snake venom action: are enzymes involved in it?

Enzymes were the first clearly recognized components of snake venoms. When several more were discovered, attempts were made to correlate venom action with enzymic functions. The last few years have seen most successful efforts in the identification, isolation and structrual elucidation of highly toxic polypeptides present in snake venoms, in particular of 'neurotoxins' and membrane-active toxins. Following this development the polypeptides were called the true toxic components and the enzymes lost their previous central position in venom pharmacology. The time, therefore, has come re-evaluate the role of enzymes in the complex interaction between snake and prey. While highly active polypeptides indeed dominate the actionof hydrophiid venoms, they appear to play a lesser role in crotalid venom action as compared with enzyme components. Enzymes are involved in many levels of venom action, e.g. by serving as spreading factors, of by producing very active agents, such as bradykinin and lysolecithins in tissues of preys or predators. Some toxins, e.g. the membrane-active polypeptides appear to participate in the interaction between membrane phospholipids and venom phospholipases. The classical neurotoxin, beta-bungarotoxin, has been recognized as a powerful phospholipase. Several instances are known which indicate that some enzymes potentiate the toxic action of others; the analysis of a single enzyme may, therefore, not fully reveal its biofunction. For 3 enzymes,ophidian L-amino acid oxicase, ATPpyrophosphatase, and acetylcholinesterase, some of the problems pertaining to venom toxicity are discussed.     

Snake venom action: Are enzymes involved in it?

Summary Enzymes were the first clearly recognized components of snake venoms. When several more were discovered, attempts were made to correlate venom action with enzymic functions. The last few years have seen most successful efforts in the identification, isolation and structural elucidation of highly toxic polypeptides present in snake venoms, in particular of neurotoxins and membrane-active toxins. Following this development the polypeptides were called the true toxic components and the enzymes lost their previous central position in venom pharmacology. The time, therefore, has come to re-evaluate the role of enzymes in the complex interaction between snake and prey. While highly active polypeptides indeed dominate the action of hydrophiid venoms, they appear to play a lesser role in crotalid venom action as compared with enzyme components. Enzymes are involved in many levels of venom action, e. g. by serving as spreading factors, of by producing very active agents, such as bradykinin and lysolecithins in tissues of preys or predators. Some toxins, e. g. the membrane-active polypeptides appear to participate in the interaction between membrane phospholipids and venom phospholipases. The classical neurotoxin, -bungarotoxin, has been recognized as a powerful phospholipase. Several instances are known which indicate that some enzymes potentiate the toxic action of others; the analysis of a single enzyme may, therefore, not fully reveal its biofunction. For 3 enzymes, ophidianl-amino acid oxidase, ATPpyrophosphatase, and acetylcholinesterase, some of the problems pertaining to venom toxicity are discussed.     

The isolation,identification and synthesis of the alarm pheromone ofVespula squamosa (Drury) (Hymenoptera: Vespidae) and associated behavior

Summary A material that elicits alarm and attack behavior byVespula squamosa (Drury) workers was isolated from venom extracts and identified by spectroscopic methods as N-3-methylbutylacetamide. This compound elicited attack responses from worker wasps identical to those responses observed when venom was applied at the same dosage. This is the first behavioral role reported for this compound.The authors thank E. Adamak, R. Murphy and F. Takken for technical assistance. This article reports the results of research only. Mention of a proprietary product does not constitute an endorsement or the recommendation for its use by USDA.     

Notes on toxinology

Conclusions Some of the papers to follow in the present series of communications on toxinology will show that in the field of chemistry, physiology, pharmacology and immunology, as well as molecular biology, animal venoms provide us with some particularly useful models. Obviously, this is one of the main reasons for the growing interest shown by numerous scientists in animal toxins.With reference to medicine, more research is needed in the field just mentioned with the aim of improving medical care. In addition, however, it is postulated that research on the behaviour of venomous animals towards man, and research into the quantities of venom actually applied to man, be intensified. Also, on the basis of results in this context, people most exposed could be provided with more and better information about prevention.Work of this sort requests the collaboration of biologists, who observe toxic animals in their natural habitat and who investigate in particular when, and under what prerogatives, the animals make use of toxins in their natural surroundings. Thus we end up with what has been said in the introduction to these notes: toxic animals are to be studied as entities and toxicity has to be looked at from all aspects essential for life, possibly including parasite and population control.Should the very last point prove valid, fascinating links could be established between toxinology and ecology and in turn migt become important for nature conservation. Thus, toxinology is but a budding field, the limits of which, can yet only be assumed.     

Plasma aldosterone in newborn and adultVipera aspis

Summary In the venomous viviparous snakeVipera aspis, plasma aldosterone concentration shows significant seasonal changes mainly related to temperature-dependent behavior. A difference is also present between newborns and adults. A correlation between sodium and potassium status and aldosterone plasma level in active and inactive life is suggested.     

Snake venom components and their applications in biomedicine

Snake envenomation is a socio-medical problem of considerable magnitude. About 2.5 million people are bitten by snakes annually, more than 100,000 fatally. However, although bites can be deadly, snake venom is a natural biological resource that contains several components of potential therapeutic value. Venom has been used in the treatment of a variety of pathophysiological conditions in Ayurveda, homeopathy and folk medicine. With the advent of biotechnology, the efficacy of such treatments has been substantiated by purifying components of venom and delineating their therapeutic properties. This review will focus on certain snake venom components and their applications in health and disease. Received 6 July 2006; received after revision 14 August 2006; accepted 28 September 2006