Effects of the insecticide ß-endosulfan on tadpoles of Isthmohyla pseudopuma (Anura: Hylidae).

Conventional agriculture uses pesticides intensively. Once pesticides are released into the environment, they can be toxic to non-target organisms. Exposure of amphibians to pesticides can be lethal and affect their growth, development and behavior. ß-endosulfan is a persistent organochlorine that has been detected in environmental samples within protected sites in Costa Rica, far from agricultural areas. The aim of this study was to evaluate the lethal and sublethal effects, as well as changes in three biomarkers (Cholinesterase activity [ChE], glutathione S-transferase activity [GST] and lipid peroxidation [LPO]) in tadpoles of Isthmohyla pseudopuma exposed to ß-endosulfan. A 96-h acute test (20, 40, 60, 80, 100 and 200 µg/L) was performed in order to calculate the median lethal concentration (LC50), while effects on growth and development were assessed during a 4-weeks chronic test (10, 20, 30 and 50 µg/L). In addition, we measured the aforementioned biomarkers in tadpoles exposed to concentrations below the LC50. The 96-h LC50 for this species was 123.6 µg/L. We found no evidence of ß-endosulfan influencing any of the three biomarkers evaluated. At 50 µg/L, both length and total weight of tadpoles decreased with respect to the control. Also, at 30 and 50 µg/L we observed that individuals showed a slower development. Therefore, we demonstrated that at sublethal concentrations, ß-endosulfan negatively affects I. pseudopuma at early stages causing tadpoles to develop slower and smaller than normal.

Health status of Polychrus gutturosus based on physical examination, hematology and biochemistry parameters in Costa Rica.

Studies evaluating the health status and characteristics of free-ranging wildlife populations are scarce or absent for most species. Saurian health assessments are usually performed in species that have conservation issues or that are kept in captivity. The Berthold's bush anole (Polychrus guturossus) is one of eight species belonging to the genus Polychrus, the only representative of the family Polychrotidae. Only a handful of studies have been reported concerning these lizard's morphological variation, ecology, and natural history, probably because P. gutturosus is a canopy dweller and it can be difficult to locate individuals. It is believed that deforestation and habitat modification could pose a threat for this species, although to date no health assessment has been done. The aim of this study was to generate health baseline data on P. gutturosus. Forty Berthold's bush anoles (20 males and 20 females) were sampled at the Pacific versant in Costa Rica, where physical examination, skin and cloacal temperatures, and blood samples were obtained from individuals immediately after capture. Animals from the studied population were all healthy (body condition 2.5-3.0/5.0). No lesions or ectoparasites were detected, but hemoparasites were found in nine individuals. Hematological and biochemical values were obtained, and the morphology of leukocytes were found to be similar to other iguanians. A positive correlation was found between the tissue enzymes aspartate aminotransferase (AST) and creatinine kinase (CK) and a negative correlation was found between skin and cloacal temperatures and AST and CK. There were positive correlations between female weight and total protein, calcium, and the calcium and phosphorus ratio. No significant inter-sex differences were found in biochemical values, despite females being larger than males. This is the first health assessment performed on a free-ranging canopy dwelling lizard. These findings provide baseline data that may be useful for future monitoring if the species faces changes in health status due to anthropogenic causes or natural disturbances.

Differences in plasmatic butyrylcholinesterases (BChE) values between Pacific and Caribbean populations of terciopelo (Bothrops asper) in Costa Rica.

The terciopelo (Bothrops asper) inhabits human-modified environments such as agricultural areas, becoming more prone to be exposed to organophosphate insecticides. These chemicals can inhibit plasmatic butyrylcholinesterases (BChE) activity in B. asper. Caribbean and Pacific populations of B. asper belong to two divergent lineages that exhibit robust genetic partitioning; however, differences across versants in biochemistry and hematology have not yet been examined, especially in BChE variations. This study aims to evaluate the differences of BChE plasmatic values, hematology parameters, and some biochemical analytes as biomarkers in the presence of organophosphates between the Pacific and Caribbean population of B. asper in Costa Rica. A total of 89 snakes (41 Pacific and 48 Caribbean) were used, and hematology parameter, albumin, aspartate aminotransferase (AST), total protein and BChEs were evaluated. Differences in hemoglobin content, thrombocytes, white cell count, AST, and BChE values were found between both versants. Intrinsic genetic factors might influence the variation found in BChE and AST values in the snakes sampled from both versants; moreover, understanding this variation in BChE and AST values across the B. asper's distribution can be useful in future ecotoxicology, biomonitoring, genetic and other clinical/health studies.

Effect of exposition to chlorpyrifos upon plasmatic cholinesterases, hematology and blood biochemistry values in Bothrops asper (Serpentes: Viperidae).

The terciopelo (Bothrops asper), is one of the most important venomous snakes in Costa Rica and common on agriculture where insecticides are frequently used for pest control. To assess the exposure to organophosphates on captive B. asper, an experiment using chlorpyrifos and butyrylcholinesterase (BChE), as a biomarker was conducted. In addition to BChE, hematology, aspartate aminotransferase (AST), total proteins (TP) and albumin were measured after exposure. Different concentrations of chlorpyrifos were used in Group A (0.1%) and B (1%), while the Control Group received distilled water; each group was composed of 5 snakes. Values of BChE, AST, TP, and albumin were measured before exposure, and at 6, 12, 24, 196, 360 and 528 h post-exposure. Hematology values were measured after 24 h post-exposure. As result, an important variation between subjects in all groups before exposure was obtained. Moreover, BChE activity showed 37% inhibition of Group A when compared to Control Group at 12 h post-exposure, and a higher inhibition of Group B (97%) related to Control Group, at 6 h post-exposure. Recovery of BChE occurred towards 528 h, never reaching initial values. Despite some variation in the rest of parameters used, a marked relative lymphopenia and monocytosis occurred at 24 h, assuming stress as the main cause.

A multi-locus molecular phylogeny for Australia's iconic Jacky Dragon (Agamidae: Amphibolurus muricatus): phylogeographic structure along the Great Dividing Range of south-eastern Australia.

Jacky dragons (Amphibolurus muricatus) are ubiquitous in south-eastern Australia and were one of the first Australian reptiles to be formally described. Because they are so common, Jacky dragons are widely used as a model system for research in evolutionary biology and ecology. In addition, their distribution along the Great Dividing Range of eastern Australia provides an opportunity to examine the influence of past biogeographical processes, particularly the expansion and contraction of forest habitats, on the diversification of this iconic agamid lizard. We generated sequence data for two mitochondrial and three nuclear DNA loci (4251base pairs) for 62 Jacky dragons sampled from throughout their distribution. Phylogenetic analyses based on maximum likelihood and Bayesian species-tree methods revealed five geographically structured clades separated by up to 6% mitochondrial and 0.7% nuclear sequence divergence. We also quantified body proportion variation within and between these genetic clades for more than 500 specimens and found no evidence of any significant differentiation in body proportions across their range. Based on body proportion homogeneity and lack of resolution in the nuclear loci, we do not support taxonomic recognition of any of the mitochondrial clades. Instead, A. muricatus is best thought of as a single species with phylogeographic structure. The genetic patterns observed in the Jacky dragon are consistent with fragmented populations reduced to multiple refugia during cold, arid phases when forested habitats were greatly restricted. Consequently, the inferred biogeographic barriers for this taxon appear to be in line with lowland breaks in the mountain ranges. Our results are congruent with studies of other reptiles, frogs, mammals, birds and invertebrates, and together highlight the overarching effects of widespread climatic and habitat fluctuations along the Great Dividing Range since the Pliocene.