Reproductive cycle of the Neotropical Crotalus durissus terrificus: I. Seasonal levels and interplay between steroid hormones and vasotocinase.

Crotaline snakes present delayed fertilization and sperm storage because secondary vitellogenesis is not completed by the time of mating. The release of vitellogenesis and synchrony between ovulation and fertilization suggest a steroidal modulation. We investigated changes of sexual steroid levels during reproduction in the Neotropical rattlesnake Crotalus durissus terrificus, analyzing macroscopical variations of reproductive condition (vitellogenesis, pregnancy, and post-partum) and plasma levels of estradiol, progesterone, and vasotocinase cystine aminopeptidase (CAP) activity over 2 years. Data showed 44.4% non-reproductive snakes (40.1% primary vitellogenesis and 4.3% post-partum) and 55.6% reproductive (36.8% secondary vitellogenesis and 18.8% pregnant). Estradiol was low in spring and summer, increasing in autumn till it peaked in winter. Estradiol in secondary vitellogenesis was significantly higher than in primary vitellogenesis, or in pregnant and post-partum females, Progesterone dropped significantly in autumn compared to summer, winter, and spring. Pregnant females showed the highest levels of progesterone compared to primary or secondary vitellogenesis, or post-partum females. CAP activity showed lowest values in reproductive females in autumn and greatest levels in post-partum females. A significant negative linear relationship was obtained between CAP activity and estradiol. The combination of morphological observations, levels of steroids and CAP activity allowed us to suggest a similar morphological reproductive pattern between temperate and tropical rattlesnakes, and to infer the role of estradiol, progesterone and CAP activity on vitellogenesis, gestation and sperm storage, respectively.

Reproductive cycle of the Neotropical Crotalus durissus terrificus: II. Establishment and maintenance of the uterine muscular twisting, a strategy for long-term sperm storage.

Crotaline snakes store sperm by means of a uterine musculature twisting (UMT). We investigated the influence of plasma levels of estradiol and progesterone and vasotocinase cystine aminopeptidase (CAP) activity on UMT formation and maintenance, and the in vitro uterine reactivity for AVT in Crotalus durissus terrificus in primary or secondary vitellogenesis with or without UMT. Frequency of females in secondary vitellogenesis with UMT is significantly higher than in primary one. Estradiol levels did not vary in all conditions studied, however, significantly low levels of progesterone were found in snakes in secondary vitellogenesis with UMT compared to those without it. UMT is always observed when high levels of estradiol and low levels of progesterone are detected. CAP activity did not change in the presence of UMT. AVT produced concentration-response contractions of the isolated uterus of snakes in all stages analysed and the pD2 value and maximum contractile response were significantly higher in primary vitellogenesis without UMT than in other reproductive conditions, indicating that uterus of those snakes presents a higher contractile capacity which may favour UMT establishment. In conclusion, we show a relationship of UMT and estradiol/progesterone balance and a possible participation of AVT in UMT formation and maintenance in the Neotropical rattlesnake.

Hemorrhagic, fibrinogenolytic and edema-forming activities of the venom of the colubrid snake Philodryas olfersii (green snake).

The venom of P. olfersii has high hemorrhagic, edema-inducing and fibrin(ogen)olytic activities. It is devoid of thrombin-like, procoagulant, phospholipase A2 and platelet aggregating enzymes. The main activities are metalloproteinases inhibited by metal chelators (EDTA and 1,10-phenanthroline) and sulfhydryl compounds (DTT and cysteine). The hemorrhagic and fibrinogenolytic enzymes were partially purified by gel filtration on HPLC. The hemorrhagic activity of the venom was neutralized by commercial horse antivenoms to Bothrops species, as well as by rabbit antisera specific for hemorrhagic factors isolated from these Bothrops venoms. No immunoprecipitin reactions were obtained, indicating that the few epitopes of the P. olfersii hemorrhagin are involved in these neutralization reactions. The fibrinogenolytic enzyme cleaves A alpha-chain more quickly than the B beta-chain of human fibrinogen. The venom also solubilizes fibrin. This solubilization appears to occur from the hydrolysis of unpolymerized alpha-chain and cross-linked gamma-gamma dimer. The fibrin peptide products are distinct from those produced by plasmin.