Basic and Applied Herpetology

Effects of alpha-cypermethrin and difenoconazole on survival, growth and biomarkers in European green toad tadpoles (Bufotes viridis, Laurenti 1768)

Bruno Bekić — 2024-12-17

In recent decades, the decline of amphibian populations has become more pronounced and accelerated, with one of the contributing factors being the excessive use of pesticides. Alpha-cypermethrin, an insecticide, and difenoconazole, a fungicide, are widely employed pesticides globally. To gain a comprehensive understanding of the acute impact of these two pesticides on amphibians, we used European green toad (Bufotes viridis) tadpoles as our model organisms. We assessed mortality and mass as apical endpoints, and examined physiological biomarkers including electron transport system (ETS) activity, catalase activity, and carbonyl protein content. Tadpoles were exposed to varying concentrations of the two pesticides for 48 hours (0.2-10 µg/L for alpha-cypermethrin; 10-100 µg/L for difenoconazole). Our results indicate that the exposure of tadpoles to these pesticides significantly affected their physiology. Mortality was observed exclusively in tadpoles treated with difenoconazole (LC₅₀ = 100.832 µg/L), while a decrease in mass occurred in tadpoles exposed to both pesticides. No significant differences across treatments were found for total ETS activity, catalase activity, or the presence of protein carbonyls. In conclusion, our findings suggest that alpha-cypermethrin at environmentally relevant concentrations poses a risk to European green toad tadpoles, while the effects of difenoconazole are become patent at higher concentrations than those recorded in natural aquatic environments.

Testudo graeca tripolitania, a new taxon of spur-thighed tortoise found in Libya at risk of exploitation for the international pet trade

Stephen F. Poterala — 2024-12-16

The spur-thighed tortoise, Testudo graeca, is a widely distributed and deeply diversified species inhabiting areas of Africa, Asia, and Europe. For decades, populations of T. graeca from North Africa have been exploited for the international pet market. In particular, T. graeca of Libyan origin have been commercially imported into the United States several times since 2021. Using mitochondrial DNA (mtDNA) sequencing, we show that these imported tortoises include T. g. cyrenaica and a novel lineage originally reported in 2017 from a displaced specimen from a market in Libya. That novel lineage inhabits northwestern Libya, and we show that wild populations near Gharyan share the same mtDNA haplotype as those now sold as pets in the USA. Populations of T. graeca in northwestern Libya, previously identified as T. g. nabeulensis, are reassigned to a new subspecies Testudo graeca tripolitania described herein. Additional field work is needed to determine the mtDNA haplotypes of populations in central and southern Tunisia and to locate the subspecies boundary between T. g. nabeulensis and T. g. tripolitania.

The Mesoamerican giant toad (Rhinella horribilis) as bioindicator of vegetation degradation in a tropical forest

Carmen Duque-Amado — 2024-06-12

Identifying species that can serve as bioindicators of environmental quality is essential for monitoring the anthropogenic impact. Common and widespread species can be ideal bioindicators due to their abundance and easy monitoring, but a confirmation of their differential responses as a function of habitat perturbation is needed. Because amphibians are known as good bioindicators of environmental perturbation, we conducted this work to identify whether a common, generalist amphibian species, the Mesoamerican giant toad (Rhinella horribilis), could serve as a bioindicator of environmental degradation in a tropical forest. We sampled toads in two areas of tropical forest that differed in anthropogenic degradation (primary vs. secondary forest), establishing in each of these areas two sections of the same surface area but differing in substrate (grass vs. sand). We analyzed toad abundance, sex ratio, body length and condition, and the amount and distribution across the body of ectoparasites (ticks). We analyzed 59 toads that were infested with 503 ticks. Based on a multi-model inference approach, the results suggested that toads were more abundant and had lower body condition in the secondary than in the primary forest. In the secondary forest, females were proportionally less abundant than males. The tick loads responded to an interaction of the body area with either the forest type or the substrate, with increased occurrence of ticks in toads from secondary forests and from grass sections. The differences found between the primary and secondary forests in sex ratio, toad abundance, body condition, and tick load across body regions are consistent with previous studies in other less common species of amphibians and thus posit R. horribilis as a good bioindicator of anthropic disturbance in this tropical forest.

High prevalence of Trypanosoma infection in Iberian green frogs (Pelophylax perezi) and evidence of a negative relationship between blood parasites and two indices of frog body condition

Rodrigo Megía-Palma — 2024-12-17

Trypanosoma commonly parasitizes anuran hosts but very few studies have investigated ecological relationships in multiparasitized amphibians. We analysed a sample of 29 adult Iberian green frogs (Pelophylax perezi) from a monitored population in central Spain and found that 28 of these individuals (96.5%) were infected with blood parasites. The protozoa genera Lankesterella (Apicomplexa: Eimeriorina) (72.4%) and Trypanosoma (Euglenozoa: Trypanosomatida) (69%) had the highest prevalence, followed by an intraerythrocytic bacteria of the genus Aegyptianella (Pseudomonadota: Rickettsiales) (31.0%). We also report an infection by hematic microfilariae (Nematoda: Spirurida) (6.8%), which to our knowledge represents the first documented case in Iberian amphibians. Infections with more than one parasite type occurred in 62.1% of the frogs. A multimodel inference approach indicated that the infection intensities of Trypanosoma and Aegyptianella were the most important predictors, both negatively affecting the body condition of the frogs. Furthermore, the number of leeches that frogs had when captured showed a strong positive association with Trypanosoma infection intensity. This suggests that leeches act as primary vectors for Trypanosoma. Our results revealed a high taxonomic diversity of blood parasites in green frogs, thus contributing to expand our knowledge of the biodiversity of Mediterranean wetlands and highlighted the potential negative effects of certain infections on the fitness of these amphibian hosts.

What can the location of skin sheds tell us? A case study on Montpellier snake Malpolon monspessulanus in southeast France

Marc Cheylan — 2024-07-30

Montpellier snake Malpolon monspessulanus (Hermann, 1804) skin sheds were collected over four years from an olive grove (0.21 ha) in southeast France. Examination of the cephalic scales allowed us to identify the individuals that frequented the site and the regularity of their visits for shedding. Reconstruction of the life histories of the individuals by capture-mark-recapture modelling showed that they visited the site during four consecutive years to shed. This site fidelity could be linked to mating opportunities or egg laying and foraging, or to the distinctive nature of the site, which is a sunny clearing within a dense woodland not very suitable for this thermophilic species. The concentration of the population in a very small area at certain times of the year risks making it vulnerable to predators as well as to changes that the site may undergo in the future, particularly if its agricultural use is abandoned and the woodland encroaches the open field area.

Barred, blotched or intermediate? A photographic-based survey on the spatial distribution of Vipera aspis phenotypes from central-southern Italy

Matteo Riccardo Di Nicola — 2024-06-12

Variation in animal colouration, which holds ecological and evolutionary significance, is evident in snakes, including the polytypic and highly polymorphic asp viper Vipera aspis. This species, currently acknowledged with four subspecies, features three of them (V. a. aspis, V. a. francisciredi and V. a. hugyi) within the Italian territory, each displaying distinct dorsal patterns. Analysing 143 georeferenced photos from personal surveys and citizen science, the present study explores transition areas between V. a. francisciredi and V. a. hugyi, revealing the coexistence of three dorsal pattern phenotypes (i.e. francisciredi, hugyi, and intermediate) across central-southern Italy. The outcome of the work reveals occurrences of francisciredi phenotypes considerably beyond the southern limit indicated in recent literature. Remarkably, intermediate phenotypes cover a significant portion of southern Italy, including Apulia (three records), Basilicata (six records) and Campania (three records) regions, and overlapping with both hugyi and francisciredi ranges. These findings suggest asymmetrical gene flow from northern to southern populations, resembling patterns observed in other reptiles. The intricate distribution of colour phenotypes underscores the necessity for molecular investigations to comprehend the genetic context and evolutionary history of V. aspis populations from central-southern Italy.

Reduction of amphibian roadkill by one-side barriers

João Rato — 2024-04-05

Amphibians are the most affected vertebrates by roadkill. Often, to minimise this problem mitigation barriers are installed. While two-side barriers effectiveness is well described in the literature, one-side barriers are poorly studied. The present study aimed to evaluate the effectiveness of one-side barriers installed by LIFE LINES project along the EM535 road in south-east Portugal. Two types of one-side barriers (one permanent and another temporary one made from tarp) were monitored through road surveys. For each barrier, the influence of interaction between the period (before and after the barrier installation) and road sector (control-no barrier, permanent or temporary barrier installed) on the number of amphibians was evaluated. The result showed that, contrary to the temporary barrier, the permanent one-side barrier appeared to be effective. Thus, the ineffectiveness of the temporary barrier may be related to (i) the tarp material that allows some amphibians to escape, (ii) the presence of vegetation that goes over the top of the barrier, or (iii) increased fence-end effects due to its short length. Even though less effective than two-side barriers, one-side barriers could be installed when there is budget limitation in conservation projects or if land use conflicts exist. Given that our study was the first to evaluate the effectiveness of one-side barriers, we discuss the potential constrains that may have affected our results.

Comparative acute toxicity of pesticides to tadpoles of a tropical anuran (Epipedobates anthonyi), a North American native anuran (Lithobates sphenocephalus) and a standard fish species

Scott Weir — 2024-02-19

Global amphibian declines have the highest incidence in tropical regions, but most of the ecotoxicological data on amphibians is collected on temperate northern hemisphere anuran species. We tested the hypothesis that tropical anuran larvae (Epipedobates anthonyi) would be more sensitive to pesticides than a North American native species (Lithobates sphenocephalus). For 12 pesticides, 96-hr range-finding acute toxicity tests were conducted to determine if mortality occurred at environmentally relevant levels. Based on those studies, two substances were selected for additional time-to-event analyses in both species as well as median lethal concentration (LC50) calculations. Time-to-event results indicated that the two species appear to be roughly equivalent in their sensitivity to the two tested pesticides. Significant differences between species were not consistent across concentrations for either the insecticide terbufos or the herbicide pendimethalin. The utility of LC50 data was mixed with one LC50 providing an arbitrarily large standard error around the LC50 precluding informative comparisons across species. However, standard LC50 methods allowed data collection that continues to contribute to our understanding of the protectiveness of fish as surrogates for anuran larvae. While our data set is limited, it appears that testing temperate species would be protective for tropical species in ecological risk assessments. Our data also support the continued use of fish as surrogates for amphibian larvae as none of the species were more sensitive to the tested pesticides than rainbow trout (Oncorhynchus mykiss), the standard sensitive fish species used for acute toxicity testing.

Is the Pyrenean newt (Calotriton asper) a thermoconformer? Cloacal and water temperature in two different thermal periods in a Pre-Pyrenean stream population

Albert Montori — 2024-02-14

In ectothermic animals, heat seems to be a determining factor because it influences many vital activities such as locomotion, the ability to escape, feeding, and reproduction, among others. In aquatic environments, physical characteristics of water prevent small ectotherms from thermoregulating and therefore it is expected that their body temperature remains similar to water temperature. Throughout its distribution and annual cycle, the Pyrenean newt (Calotriton asper) is exposed to a wide range of water temperatures that affect its biological and ecological traits like the standard metabolic rate, oxygen consumption, activity period and growth pattern. This study analyses the relationship between the cloacal and water temperatures in a Pre-Pyrenean population of C. asper in two periods with well differentiated water temperatures (July and September). The aims are to establish if there are differences between sexes in cloacal temperature, whether reproductive activity modifies cloacal temperature, and to analyse the degree of thermoconformity of the species. The results indicate that cloacal temperature depends on the water temperature, corroborating the idea that C. asper is mainly a thermoconformer, as it corresponds to an aquatic ectotherm. However, the cloacal temperature of newts was slightly higher than water temperature. In individuals in amplexus, cloacal temperature was significantly higher than in specimens that are not in this mating position, and in July, females showed much greater dispersion in cloacal temperature than males, which is probably related to spawning behaviour.

Species list of the Spanish herpetofauna: an update

Alberto Sánchez-Vialas — 2024-12-20

Since the last update of the list of the Spanish herpetofauna in 2018, recent studies have provided new evidence supporting the need to implement taxonomic changes in several groups. In this work, we present an updated reference list for the Spanish herpetofauna, which currently includes 132 native or historically introduced species, i.e. 37 amphibians and 95 reptiles, offering a standardized framework for amateurs, scientists, and environmental agencies. While updating the list, we had to undertake some nomenclatural acts. For instance, to solve the nomenclatural issues concerning Blanus species, we designate a neotype for Amphisbaena oxyura Wagler, 1824; thus, the valid name for the two recognized Iberian species are Blanus cinereus and Blanus oxyurus. We also provide an updated list for the 13 introduced species with documented breeding populations in Spain.