The time course of acclimation of critical thermal maxima is modulated by the magnitude of temperature change and thermal daily fluctuations.

Plasticity in the critical thermal maximum (CTmax) helps ectotherms survive in variable thermal conditions. Yet, little is known about the environmental mechanisms modulating its time course. We used the larvae of three neotropical anurans (Boana platanera, Engystomops pustulosus and Rhinella horribilis) to test whether the magnitude of temperature changes and the existence of fluctuations in the thermal environment affected both the amount of change in CTmax and its acclimation rate (i.e., its time course). For that, we transferred tadpoles from a pre-treatment temperature (23 °C, constant) to two different water temperatures: mean (28 °C) and hot (33 °C), crossed with constant and daily fluctuating thermal regimes, and recorded CTmax values, daily during six days. We modeled changes in CTmax as an asymptotic function of time, temperature, and the daily thermal fluctuation. The fitted function provided the asymptotic CTmax value (CTmax∞) and CTmax acclimation rate (k). Tadpoles achieved their CTmax∞ between one and three days. Transferring tadpoles to the hot treatment generated higher CTmax∞ at earlier times, inducing faster acclimation rates in tadpoles. In contrast, thermal fluctuations equally led to higher CTmax∞ values but tadpoles required longer times to achieve CTmax∞ (i.e., slower acclimation rates). These thermal treatments interacted differently with the studied species. In general, the thermal generalist Rhinella horribilis showed the most plastic acclimation rates whereas the ephemeral-pond breeder Engystomops pustulosus, more exposed to heat peaks during larval development, showed less plastic (i.e., canalized) acclimation rates. Further comparative studies of the time course of CTmax acclimation should help to disentangle the complex interplay between the thermal environment and species ecology, to understand how tadpoles acclimate to heat stress.

The effect of thermal microenvironment in upper thermal tolerance plasticity in tropical tadpoles. Implications for vulnerability to climate warming.

Current climate change is generating accelerated increase in extreme heat events and organismal plastic adjustments in upper thermal tolerances, (critical thermal maximum -CTmax ) are recognized as the quicker mitigating mechanisms. However, current research casts doubt on the actual mitigating role of thermal acclimation to face heat impacts, due to its low magnitude and weak environmental signal. Here, we examined these drawbacks by first estimating maximum extent of thermal acclimation by examining known sources of variation affecting CTmax expression, such as daily thermal fluctuation and heating rates. Second, we examined whether the magnitude and pattern of CTmax plasticity is dependent of the thermal environment by comparing the acclimation responses of six species of tropical amphibian tadpoles inhabiting thermally contrasting open and shade habitats and, finally, estimating their warming tolerances (WT = CTmax - maximum temperatures) as estimator of heating risk. We found that plastic CTmax responses are improved in tadpoles exposed to fluctuating daily regimens. Slow heating rates implying longer duration assays determined a contrasting pattern in CTmax plastic expression, depending on species environment. Shade habitat species suffer a decline in CTmax whereas open habitat tadpoles greatly increase it, suggesting an adaptive differential ability of hot exposed species to quick hardening adjustments. Open habitat tadpoles although overall acclimate more than shade habitat species, cannot capitalize this beneficial increase in CTmax, because the maximum ambient temperatures are very close to their critical limits, and this increase may not be large enough to reduce acute heat stress under the ongoing global warming.

Toxinological characterization of venom from Leptodeira annulata (Banded cat-eyed snake; Dipsadidae, Imantodini).

We investigated the histology of Duvernoy's venom gland and the biochemical and biological activities of Leptodeira annulata snake venom. The venom gland had a lobular organization, with secretory tubules formed by serous epithelial cells surrounding each lobular duct. The latter drained into a common lobular duct and subsequently into a central cistern. In contrast, the supralabial gland was mucous in nature. SDS-PAGE revealed a profile of venom components that differed from pitviper (Bothrops spp.) venoms. RP-HPLC also revealed greater complexity of this venom compared to Bothrops venoms. The venom had no esterase, l-amino acid oxidase or thrombin-like activity, but was proteolytic towards elastin-Congo red, fibrin, fibrinogen, gelatin and hide powder azure. The venom showed strong α-fibrinogenase and fibrinolytic activities and reduced the rate and extent of plasma recalcification. The proteolytic activity was inhibited by EDTA and 1,10-phenanthroline (metalloproteinase inhibitors) but not by AEBSF and PMSF (serine proteinase inhibitors). The venom had phospholipase A2 (PLA2) activity that was inhibited by varespladib. The venom cross-reacted with antivenoms to lancehead (Bothrops spp.), coralsnake (Micrurus spp.) and rattlesnake (Crotalus durissus terrificus) venoms. The venom did not aggregate rat platelets or inhibit collagen-induced aggregation, but partially inhibited thrombin-induced aggregation. The venom was hemorrhagic (inhibited by EDTA) and increased the vascular permeability (inhibited by varespladib) in rat dorsal skin. In gastrocnemius muscle, the venom caused myonecrosis and increased serum creatine kinase concentrations. In conclusion, L. annulata venom has various enzymatic and biological activities, with the local effects being mediated primarily by metalloproteinases and PLA2.