Furthermore, the results Selleck AC220 from the individual chromatograms (Fig. 1B) of the venoms clearly demonstrates the presence of isoforms of crotamine, crotapotin and phospholipase A2, of the crotoxin complex. The phospholipase A2 isoforms were found in more abundance, being observed in the majority of the chromatograms of the studied groups. Despite the
high variability in the concentrations, the chromatographic profiles did not present variation of venom proteins when considering the captivity time and ontogenetic variation. The RP-HPLC profile of the crotamine-positive and -negative venoms can be observed in Fig. 2A and B respectively. The contents of major peaks were determined by manual collection of the fraction followed by Edman degradation. In the case of crotapotin, the peptide was reduced with Dithiothreitol and alkylated with iodoacetamide (following standard protocols) and resubmitted to another chromatographic separation before individual sequencing of the peaks. The results on toxicity and coagulant activity are presented in Table 2 that shows LD50 statistical difference between males and females wild life groups. Statistical difference was observed also for clotting BIBF 1120 mouse time (CT) to newborns and reference venom group. The group of animals
inoculated with crotamine-positive venom presented hypertonicity of the hind paws followed by complete paralysis. The groups inoculated with crotamine-negative venom and control did not present any neurological activities. The monitoring of biological, biochemical and pharmacological activities of venoms should be one of the great concerns of institutions that produce antivenom, given that studies conducted in the last 50 years have demonstrated variation in these activities attributable
to sex, Linifanib (ABT-869) age range, geographic origin, diet, captivity, season of the year and/or possible environmental changes (Ferreira et al., 2009, 2010a, in press; Campagner et al., in press; Schenberg, 1959b; Furtado et al., 2003; Pimenta et al., 2007; Calvete et al., 2009). This variability has direct implications on the antivenom type produced and, depending on the favorable response or lack thereof, on the treatment of snakebite patients (Chippaux et al., 1991; Warrel, 1997; Calvete et al., 2009). In the present study a large number of adult Cdt snakes (males and females) and newborns were evaluated, comparing the biological activities of animals newly born in the wild with those maintained for at least three years in captivity, finding a high variability in their venoms. Thus, the protein profile evaluated did not present a difference among the adult individuals corroborating Cárdenas et al. (1995) who found values of 76.9% in Brazilian snakes and 81.4% for Argentinean ones. On the other hand, the venom from the offspring showed a protein content of 60%, a value below that observed in adults (75%).