Variation In Venom Composition Research Articles

Comparative venom gland transcriptome surveys of the saw-scaled vipers (Viperidae: Echis) reveal substantial intra-family gene diversity and novel venom transcripts

BackgroundVenom variation occurs at all taxonomical levels and can impact significantly upon the clinical manifestations and efficacy of antivenom therapy following snakebite. Variation in snake venom composition is thought to be subject to strong natural selection as a result of adaptation towards specific diets. Members of the medically important genus Echis exhibit considerable variation in venom composition, which has been demonstrated to co-evolve with evolutionary shifts in diet. We adopt a venom gland transcriptome approach in order to investigate the diversity of toxins in the genus and elucidate the mechanisms which result in prey-specific adaptations of venom composition.ResultsVenom gland transcriptomes were created for E. pyramidum leakeyi, E. coloratus and E. carinatus sochureki by sequencing ~1000 expressed sequence tags from venom gland cDNA libraries. A standardised methodology allowed a comprehensive intra-genus comparison of the venom gland profiles to be undertaken, including the previously described E. ocellatus transcriptome. Blast annotation revealed the presence of snake venom metalloproteinases, C-type lectins, group II phopholipases A2, serine proteases, L-amino oxidases and growth factors in all transcriptomes throughout the genus. Transcripts encoding disintegrins, cysteine-rich secretory proteins and hyaluronidases were obtained from at least one, but not all, species. A representative group of novel venom transcripts exhibiting similarity to lysosomal acid lipase were identified from the E. coloratus transcriptome, whilst novel metallopeptidases exhibiting similarity to neprilysin and dipeptidyl peptidase III were identified from E. p. leakeyi and E. coloratus respectively.ConclusionThe comparison of Echis venom gland transcriptomes revealed substantial intrageneric venom variation in representations and cluster numbers of the most abundant venom toxin families. The expression profiles of established toxin groups exhibit little obvious association with venom-related adaptations to diet described from this genus. We suggest therefore that alterations in isoform diversity or transcript expression levels within the major venom protein families are likely to be responsible for prey specificity, rather than differences in the representation of entire toxin families or the recruitment of novel toxin families, although the recruitment of lysosomal acid lipase as a response to vertebrate feeding cannot be excluded. Evidence of marked intrageneric venom variation within the medically important genus Echis strongly advocates further investigations into the medical significance of venom variation in this genus and its impact upon antivenom therapy.

Interspecific variation in venom composition and toxicity of Brazilian snakes from Bothrops genus

The genus Bothrops spp. is responsible for 90% of envenomation by snakes in Brazil, and the standard treatment for snakebites is the antivenom therapy. The anti-bothropic serum produced by Butantan Institute is prepared by the hyperimmunization of horses with a pool of venoms from Bothrops alternatus, Bothrops jararaca, Bothrops jararacussu, Bothrops moojeni and Bothrops neuwiedi. In this study, the biochemical and biological characteristics of the venoms from nineteen snakes of the genus Bothrops, responsible for human accidents in Brazil, were analysed. Venoms, particularly from Crotalidae and Viperidae snakes, are rich sources of serine proteases and metalloproteases and the ability of the Brazilian anti-bothropic serum to neutralize the proteolytic activity of these venoms were also tested. The results obtained here show the existence of a large range of variation in the composition and activities in Bothrops spp. toxins and demonstrate that the anti-bothropic serum is not able to fully neutralize the toxic activities of all analysed venoms. These suggest that for the preparation of a fully effective therapeutic anti-bothropic serum, other venoms should be included in the immunization mixture.

Intersexual variations in Northern ( Missulena pruinosa) and Eastern ( M. bradleyi) mouse spider venom

Venoms of both sexes of Australian Northern ( Missulena pruinosa) and Eastern ( Missulena bradleyi) mouse spiders were studied in order to determine intersexual variations in venom yield, composition and bioactivity. Females of both species yielded more venom than males. High-performance liquid chromatography (HPLC) and mass spectrometry data further indicate a substantial degree of intersexual variation in the venom composition of both species. In a cricket ( Acheta domestica) acute toxicity assay, only small intersexual differences were observed, but M. bradleyi venom was found to be considerably more potent than M. pruinosa venom. In the chick biventer cervicis nerve–muscle preparation, male but not female M. bradleyi venom induced large and sustained muscle contractions with fasciculation and decreased twitch height that could be reversed by CSL funnel-web spider antivenom. In contrast, venoms of both sexes of M. pruinosa did not induce significant effects in the chick biventer cervicis nerve–muscle preparation. We therefore conclude that female M. bradleyi venom and venoms from male and female M. pruinosa appear to contain few, if any, orthologs of δ-missulenatoxin-Mb1a, the toxin responsible for the effects of male M. bradleyi venom in vertebrates. These findings are consistent with clinical reports that mouse spiders, particularly species other than male M. bradleyi, do not appear to be a major medical problem in humans.

Effectiveness of Snake Antivenom: Species and Regional Venom Variation and Its Clinical Impact

The ubiquity of venom variation in snakes poses special problems for the manufacture of antivenom and has undermined the commercial attractiveness of this class of therapeutic agent. In particular, it has been amply documented that both interspecific and intraspecific variation in venom composition can affect the neutralisation capacity of antivenoms. This may be exacerbated by the selective use of tests of venom toxicity

INTRASPECIFIC VARIATION IN SCORPIONLEIURUS QUINQUESTRIATUSVENOM COLLECTED FROM EGYPT (SINAI AND ASWAN DESERTS)

Leiurus quinquestriatus is a single species genus scorpion of Saharan origin, which has penetrated the Mediterranean region. The Sinai Isthmus is believed to be the region in which the subspecies L.q. quinquestriatus gives way to L.q. hebraeus. To test whether there are qualitative differences in the venom from Leiurus quinquestriatus inhabiting two different geographic regions, venom was obtained from scorpions collected from Aswan in Southern Egypt and from the Southern region of Sinai Peninsula, Egypt. Electrophoresis and a densitometric gel scan showed that in the molecular weight range above that known to include toxins, venom of Aswan origin contained several protein bands that were absent from Sinai-sourced venom. In contrast Sinai venom appeared to have a larger proportion of protein in the molecular weight range known to include toxins. Such differences may reflect a response to local ecological conditions. Application of equal concentrations by weight of the two venoms to rat ileum showed no difference in the contraction produced by the lowest effective dose (0.2 μg ml-1) but at higher doses (0.5, 1.0 & 2.0 μg ml-1) venom from Aswan induced stronger and more sustained contractions with a different time course than did Sinai scorpion venom. Geographically related intra-specific variation in venom composition might be important in treating the pathophysiological effects of scorpion venom. This is the first time that differences in the venom components and their physiological effectiveness has been demonstrated for the scorpion L. quinquestriatus venom collected from two distinct areas in Egypt.

Diet and snake venom evolution.

Venom composition within snake species can show considerable geographical variation, an important consideration because bites by conspecific populations may differ in symptomatology and require different treatments. The underlying causes of this phenomenon have never been explained. Here we present evidence that the variation in the venom of the pitviper Calloselasma rhodostoma (Serpentes: Viperidae) is closely associated with its diet. We also evaluated other possible causes of geographic variation in venom using partial Mantel tests and independent contrasts, but rejected both contemporary gene flow (estimated from geographical proximity) and the phylogenetic relationships (assessed by analysis of mitochondrial DNA) among populations as important influences upon venom evolution. As the primary function of viperid venom is to immobilize and digest prey and prey animals vary in their susceptibility to venom, we suggest that geographical variation in venom composition reflects natural selection for feeding on local prey.

Electrophoretic profiles and biological activities: Intraspecific variation in the venom of the malayan pit viper ( Calloselasma rhodostoma)

The Malayan pit viper ( Calloselasma rhodostoma) is of major clinical significance both as a leading cause of snakebite and as the source of ancrod (Arvin ™). Although its venom has been extensively studied, the degree to which venom composition varies between individuals is poorly known. We individually analysed the venoms of over 100 C. rhodostoma using isoelectric focusing. In all populations, females produced an intense band that was absent from all males, and significant ontogenetic variation was detected. Principal components analysis of the banding profiles also revealed strong geographic variation, which was significantly congruent with variation in the biological activities of the venom (phosphodiesterase, alkalinephosphoesterase, l-amino acid oxidase, arginine ester hydrolase, 5′-nucleotidase, thrombin-like enzyme, haemorrhagic activity). Studies of captive-bred snakes indicate that the intraspecific variation in venom is genetically inherited rather than environmentally induced. The intraspecific variation in venom composition and biological activity could be of applied importance to snakebite therapy, both in correct diagnosis of the source of envenomation and in the development of a more effective antivenom. Greater attention should be given to the source of C. rhodostoma venom used in research to ensure reproducibility of results.

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 identification 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 spitting 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.

Variation in venom composition and reactivity in two specimens of yellow-faced whip snake (Demansia psammophis) from the same geographical area

V. Williams and J. White. Variation in venom composition and reactivity in two specimens of yellow-faced whip snake ( Demansia psammophis) from the same geographical area. Toxicon 28, 1351–1354, 1990.—Gel filtration chromatography and SDS-PAGE of venom from two specimens of Demansia psammophis showed little similarity. Amidolytic activity of the venoms, however, was in the same order of reactivity against various chromogenic substrates. The venom from both snakes produced precipitin lines with brown snake antivenom but the venom detection kit (Commonwealth Serum Laboratories) identified one venom as brown snake ( Pseudonaja sp.) and the other as tiger snake ( Notechis sp.). These results raise questions about the phylogeny of this species.

Venom Ontogeny in the Pacific Rattlesnakes Crotalus viridis helleri and C. v. oreganus

Ontogenetic variation in venom composition was examined in the Pacific rattlesnakes Crotalus viridis helleri and C. v. oreganus. Venoms were analyzed for protease, phospholipase A2, L-amino acid oxidase, exonuclease and elastinolytic activities, and toxicity toward a native prey (Sceloporus graciosus). Protease activity increased significantly with size; L-amino acid oxidase and exonuclease activities also tended to increase. Phospholipase A2 activity decreased significantly with size, as did venom toxicity. These factors produce a highly toxic venom with low protease activity in juvenile snakes, which facilitates efficient handling of lizards and young rodents. Analysis of gut contents of museum specimens showed that lizards constitute a major fraction of prey taken by juvenile rattlesnakes. Lizards continue to be taken with high frequency until snakes reach approx. 500 mm in total length; above this size, mammals are taken exclusively. As snakes increase in size, they feed on larger mammalian prey, and a functionally different venom is produced. Venom from adult Pacific rattlesnakes is less toxic but has high protease activity, aiding in the digestion of prey in a thermally variable environment.