The emergence of disease as a significant global threat to amphibian diversity has generated considerable interest in amphibian defenses against cutaneous microbial infection and disease. To date, however, the influence of sloughing on the susceptibility of amphibians to infection and disease has been largely overlooked. To investigate the potential for sloughing to regulate topical microbial loads, the abundance of cultivable cutaneous bacteria and fungi in the cane toad Rhinella marina were compared before and after sloughing. Toads were also exposed to fluctuating thermal regimes (10−20 and 20−30°C) and variable photoperiods to investigate possible effects of season and climate on sloughing periodicity. Sloughing substantially reduced the abundance of cultivable cutaneous bacteria and fungi by up to 100%. The intermoult interval of toads maintained at 10−20°C was twice that of animals at 20−30°C and did not appear to thermally acclimate. Photoperiod had no discernable influence on sloughing periodicity. Results of this study suggest that normal sloughing cycles could play a significant role in controlling the persistence and build-up of cutaneous microbes, including pathogens. The loss of non-pathogenic commensal and protective skin microbiota after sloughing may also influence host susceptibility to cutaneous pathogens. We suggest that the spatio-temporal dynamics of chytridiomycosis, the widespread and often fatal disease caused by the fungal pathogen Batrachochytrium dendrobatidis, are related to temperature not only because of its effect on the growth of the fungus, but also because of its effect on the frequency of host sloughing.KEY WORDS: Skin · Shedding · Temperature · Rhinella marina · Pathogen · Batrachochytrium dendrobatidis · Commensal microbe · Epidermis · Cutaneous Resale or republication not permitted without written consent of the publisherDis Aquat Org 101: [235][236][237][238][239][240][241][242] 2012 and microbes adherent to the most peripheral skin layers can be shed with the shed keratinocytes (Dahl 1993). In amphibians, the stratum corneum is shed on a regular basis (Larsen 1976, Withers 1995; moreover, unlike mammals, sloughing of the stratum corneum in amphibians is usually entire (i.e. all of the stratum corneum is shed at once) (Taylor & Ewer 1956, Ling 1972, Larsen 1976, Withers 1995. As such, sloughing could play a significant role in regulating the abundance and persistence of microbes, both beneficial and pathogenic, at the body's surface.To investigate the influence of sloughing on microbial abundances on amphibian skin, we examined changes in the abundance of cultivable cutaneous microbes post-sloughing in cane toads Rhinella marina (Linnaeus, 1758). As ectotherms, physiological processes such as sloughing are likely to be affected by environmental factors such as temperature and day length. Seasonal differences in temperature and photoperiod may, therefore, affect sloughing periodicity and in turn contribute to the seasonality of some amphibian diseases (Berger et...

In the following work we examine the richness and altitudinal distribution of Colombian Andean anurans trying to emphasize patterns of distribution. We also supply an updated checklist of Andean anurans in Colombia. At present, Colombian harbors about 396 Andean frogs: 153 species in the Cordillera Occidental, 187 species in the Cordillera Central, and 131 species in the Cordillera Oriental. Of these, the Cordillera Oriental presents the higher number and percentage of endemic species. The frequency distribution of altitudinal ranges for Colombian Andean frogs shows that the majority of species have narrow altitudinal ranges, less than 500 m altitude, and only a few species have broad altitudinal distributions. On the other hand, lowland species have broader altitudinal ranges than do highland species. The hypothesis of a wider altitudinal range of highland anurans is therefore not supported. Finally, the averages of the Jaccard similarity indices for the Andean anurans along altitudinal gradients in Colombian are approximately similar to those of other tropical anurans reported by Huey (1978), but notably lower than those of anurans of temperate localities. Thus, these results are in concordance with Janzen's hypothesis (1967) about a broader altitudinal range for temperate species, likely because of their higher thermal tolerance.

The spraying of coca (Erythroxylum coca) with glyphosate (coca mixture, a combination of formulated glyphosate, Glyphos, and an adjuvant, Cosmo-Flux) in Colombia has raised concerns about possible impacts on amphibians. Although acute LC50 for 8 species of Colombian frogs ranged from 1.2 to 2.78 mg acid equivalents (a.e.)/L, these exposures were conducted in the laboratory in the absence of sediments and organic matter such as would occur under realistic field conditions. In order to assess the effects of overspray of frog habitat under field conditions, Gosner stage 25 tadpoles of Rhinella granulosa, R. marina, Hypsiboas crepitans, and Scinax ruber were placed in outdoor microcosms made from polyethylene plastic fish ponds (2.07 m in diameter, 37 cm high) in an experimental area in Tolima, Colombia. The bottoms of the microcosms were covered with a 3-cm layer of local soil and they were filled to a depth of 15 cm (above the sediment) with local spring water. After up to 100 tadpoles of each frog species were placed in the microcosms, they were sprayed with the coca mixture at concentrations greater and less than the normal application rate (3.69 kg glyphosate a.e./ha). Mortality at 96 h in the control microcosms was between 0 and 16% and LC50 values were between 8.9 and 10.9 kg glyphosate a.e./ha (equivalent to initial concentrations of 5963 to 7303 microg glyphosate a.e./L). Mortality >LC50 was only observed in the tested species when the application rate was >2-fold the normal application rate. In other experiments, juvenile and adult terrestrial stages of frogs were exposed by direct spraying to a range of concentrations of coca mixture. Juveniles and adults were exposed in plastic food containers (19 x 19 cm). The bottom of the container was filled with moistened soil and leaf litter to a depth of 1 cm and 0.5 cm, respectively. Mortality in the controls was low, from 0 to 10%, and from 0 to 35% at the normal application rate. LC50 values ranged between 4.5 kg a.e./ha and 22.8 kg a.e./ha, 1.5- to 6-fold greater than the normal application rate. Data indicate that, under realistic worst-case exposure conditions, the mixture of Glyphos and Cosmo-Flux as used for control of coca in Colombia exerts a low toxicity to aquatic and terrestrial stages of anurans and that risks to these organisms under field conditions are small.

The spraying of coca (Erythroxylum coca) with glyphosate in Colombia has raised concerns about possible impacts on amphibians. There are few toxicity data for species other than those from temperate regions, and these have not been generated with the combination of formulated glyphosate (Glyphos) and the adjuvant, Cosmo-Flux (coca mix) as used in coca control in Colombia. In order to characterize toxicity of the spray mixture to frogs from Colombia, Gosner stage-25 tadpoles of Scinax ruber, Dendropophus microcephalus, Hypsiboas crepitans, Rhinella granulosa, Rhinella marina, Rhinella typhonius, Centrolene prosoblepon, and Engystomops pustulosus were exposed to the coca mix at concentrations of glyphosate ranging from 1 to 4.2 mg a.e./L diluted in dechlorinated tap water in glass containers. Cosmo-Flux was added to Glyphos in the proportion of 2.3% v/v, as used in aerial application for coca control. Exposures were for 96 h at 23 +/- 1.5 degrees C with 12:12-h light/dark cycle. Test solutions were renewed every 24 h. Concentrations, measured within the first hour and at 24 and 96 h using enzyme-linked immunosorbent assay (ELISA) (Abraxis, LLC), ranged from 70 to 130% of nominal values. LC50 values ranged from 1200 to 2780 microg glyphosate acid equivalents (a.e.)/L for the 8 species tested. Data suggest that sensitivity to Roundup-type formulations of glyphosate in these species is similar to that observed in other tropical and temperate species. In addition, sensitivity of larval amphibians to Roundup-type formulations spans a relatively narrow range. Finally, toxicity of the mixture as used to spray coca was likely driven by the surfactant in the glyphosate formulation, as the addition of Cosmo-Flux did not enhance toxicity above those reported for Vision = Roundup.

Anuran survival is strongly affected by exposure to high environmental temperatures. However, their upper thermal tolerances vary between species and within developmental stages. The aims of this research were to measure the median lethal temperature (LT50) of three anuran developmental stages (Gosner stages 10, 20, and 25) at a constant thermal regime, and of developing embryos (stage 10) until they became tadpoles (stage 25) exposed to daily peaks of temperatures between 1000 and 1600. Four Colombian species (Emerald-eyed Treefrog, Hypsiboas crepitans (Wied-Neuwied, 1824); Tungara Frog, Engystomops pustulosus (Cope, 1864); Rivero’s Toad, Rhinella humboldti (Gallardo, 1965); Emerald Glassfrog, Espadarana prosoblepon (Boettger, 1892)) were used in these experiments. An ontogenetic increase was observed in the upper thermal tolerance from embryos to tadpoles for all species studied. In addition, developing embryos exposed to peak temperatures showed a LT50 fairly close to the mean of the maximum habitat temperatures, particularly in H. crepitans and E. pustulosus that lay egg masses exposed directly to the sun. Environmental temperatures in the microhabitat of species studied showed values remarkably higher than their experimental LT50. Therefore, we postulate that rapid increases in environmental temperatures, as result of global or local changes, might be a critical factor for anuran survival, mainly during the embryonic stages when they are more sensitive to temperature.

Temperature and humidity are critical factors for terrestrial lungless salamanders, as their body temperatures are largely determined by the environmental temperature and require moisture to sustain cutaneous respiration. Herein, we evaluated the preference of Bolitoglossa ramosi Brame and Wake, 1972 between a high temperature and a high relative humidity (RH), the influence of temperature on RH preferences, and the influence of RH on the thermal preferences. This study was performed in a field location in the municipality of Líbano, Tolima, Colombia. There, on different nights, we collected 84 adult B. ramosi and carried out the preference experiments, using aluminum troughs with different thermal and RH gradients. We found that between high temperature and high RH, B. ramosi preferred high RH. However, B. ramosi selected high temperatures when the gradient had a high RH and low temperatures when the gradient had a low RH. These results show that B. ramosi is able to thermoregulate and hydroregulate. Nevertheless, hydroregulation seems to be more important than thermoregulation because B. ramosi always selected the high RH gradients, while their thermal selection relied on the hydric environment.

Anurans are ectothermic animals very sensitive to temperature, mainly during the embryonic stage. In addition, environmental temperature decreases with altitude, and the amphibian fauna changes. Therefore, we studied the relationship between the embryonic thermal tolerances of twelve species of anurans and the temperatures of their microhabitat along an altitudinal gradient from 430 m to 2600 m. We hypothesized that there is a strong thermal adjustment of embryos to their microhabitat and, consequently, that temperature could be a limiting factor of altitudinal distribution of the anurans. We also compared the embryonic thermal tolerances according to six postulated reproductive modes of the study species. We found a significant relationship between the maximum and minimum thermal tolerances of the anuran embryos and the maximum and minimum temperatures of their microhabitat and altitudinal distribution. We also found a wide range of embryonic thermal tolerances for aquatic breeding species and a narrower range for terrestrial breeding species. Particularly, embryos of direct development species were the most sensitive to temperature. These results show the strong thermal adjustment of anuran embryos to their microhabitat and elevation and do not reject the hypothesis that temperature can be a limiting factor of their altitudinal distribution.

Understanding how geographic and environmental heterogeneity drive local patterns of genetic variation is a major goal of ecological genomics and a key question in evolutionary biology. The tropical Andes and inter‐Andean valleys are shaped by markedly heterogeneous landscapes, where species experience strong selective processes. We examined genome‐wide SNP data together with behavioural and ecological traits (mating calls and body size) known to contribute to genetic isolation in anurans in the banana tree‐dwelling frog, Boana platanera, distributed across an environmental gradient in Central Colombia (northern South America). Here, we analysed the relationships between environmentally (temperature and precipitation) associated genetic and phenotypic differentiation and the potential drivers of isolation by environment along an elevation gradient. We identified candidate SNPs associated with temperature and body size, which follow a clinal pattern of genome‐wide differentiation tightly coupled with phenotypic variation: as elevation increases, B. platanera exhibits larger body size and longer call duration with more pulses but lower pulse rate and frequency. Thus, the environmental landscape has rendered a scenario where isolation by environment and candidate loci show concordance with phenotypic divergence in this tropical frog along an elevation gradient in the Colombian Andes. Our study sets the basis for evaluating the role of temperature in the genetic structure and local adaptation in tropical treefrogs and its putative effect on life cycle (embryos, tadpoles, adults) along elevation gradients.