Hypsiboas Pulchellus Research Articles

Genotoxicity Of The Herbicide Picloram (TORDON ® 24K) on tadpoles of Boana pulchella (Amphibia, Anura, Hylidae)

Picloram (PIC) is an herbicide widely used in Uruguay to control weeds, especially in forest crops. In the present study, we evaluate the acute toxicity and genotoxicity of formulated 24.9% PIC-based Tordon®24K on Boana pulchella tadpoles under standard laboratory conditions. In the acute toxicity test we use mortality as the endpoint and we obtained a 48-h LC50 of 16.5 mg/L, LOEC 14.5 mg/L, and NOEC 12.5 mg/L. We determined genotoxicity in GS36 tadpoles exposed to sublethal concentrations of 8.5 (NOEC/1.5) and 12.5 mg/L (NOEC) PIC using micronucleus frequency (MN) and occurrence of nuclear abnormalities (AN) as endpoints. We noticed a significant increase in the frequency of MN and AN in all treatments tested at sublethal concentrations. Our findings demonstrate the genotoxic effects of this herbicide, which represent a danger to amphibians that inhabit agroecosystems. Based on the results obtained and considering that Boana pulchella is not a threatened species and has a wide distribution in the Uruguayan territory and the Pampa Biome, it could be considered a reference organism in environmental monitoring programs in the region.

Coping with dry spells: Investigating oxidative balance and metabolic responses in the subtropical tree frog Boana pulchella (Hylidae) during dehydration and rehydration exposure

In the face of climate change, understanding the metabolic responses of vulnerable animals to abiotic stressors like anurans is crucial. Water restriction and subsequent dehydration is a condition that can threaten populations and lead to species decline. This study examines metabolic variations in the subtropical frog Boana pulchella exposed to dehydration resulting in a 40% loss of body water followed by 24 h of rehydration. During dehydration, the scaled mass index decreases, and concentrations of metabolic substrates alter in the brain and liver. The activity of antioxidant enzymes increases in the muscle and heart, emphasizing the importance of catalase in the rehydration period. Glycogenesis increases in the muscle and liver, indicating a strategy to preserve tissue water through glycogen storage. These findings suggest that B. pulchella employs specific metabolic mechanisms to survive exposure to water restriction, highlighting tissue-specific variations in metabolic pathways and antioxidant defenses. These findings contribute to a deeper understanding of anuran adaptation to water stress and emphasize the importance of further research in other species to complement existing knowledge and provide physiological tools to conservation.

Biomarkers at the Individual and Biochemical Level: Effects of Pure and Formulated Lambda-Cyhalothrin in Boana pulchella Tadpoles (Duméril and Bibron, 1841).

We compared the effects of lambda-cyhalothrin as the pure active ingredient and as a formulated product (Zero®), on the larval stage of the autochthonous species Boana pulchella. We evaluated ecotoxicological endpoints, behavioral and developmental alterations, and the biochemical detoxifying, neurotoxic, and oxidative stress responses, covering a wide concentration range from environmental to high application levels. Both pyrethroid preparations displayed similar ecotoxicity (median lethal concentration of ~0.5 mg/L), with the lethal effect of Zero® being more pronounced than that of the active ingredient. Sublethal behavioral alterations in natatory activity were observed at 1000 times lower concentrations, indicating the ecological hazard of tadpole exposure to this pyrethroid at environmentally relevant concentrations. Biochemical endpoints in B. pulchella larvae showed significant responses to lambda-cyhalothrin in the ng/L range; these responses were different for the pure or the formulated product, and they were variable at higher concentrations. Principal components analysis confirmed the prevalence of biochemical responses as early endpoints at the lowest lambda-cyhalothrin concentrations; the Integrated Biomarker Response Index proportionally increased with pyrethroid concentration in a similar way for the pure and the formulated products. We conclude that lambda-cyhalothrin is of concern from an environmental perspective, with particular emphasis on autochthonous anuran development. The battery of biochemical biomarkers included in our study showed a consistent integrated biomarker response, indicating that this is a potent tool for monitoring impacts on amphibians. Environ Toxicol Chem 2024;00:1-11. © 2024 SETAC.

Attenuation Effect of Azolla spp. on Lambda-Cyhalothrin Toxicity to Aquatic Organisms.

Wetlands play a crucial role in providing valuable ecosystem services, including the removal of various pollutants. In agricultural basins, wetlands are exposed to agrochemical loads. This study aims to assess the attenuation effect of the ubiquitous macrophyte Azolla spp. on the toxicity of lambda-cyhalothrin to sensitive aquatic organisms. An indoor mesocosm experiment was conducted to compare the concentration of lambda-cyhalothrin at different time points after pesticide application in vegetated and unvegetated treatments, including a control without pesticide addition. Toxicity tests were performed throughout the experiment on three organisms: a fish (Cnesterodon decemmaculatus), a macroinvertebrate (Hyalella curvispina), and an amphibian (Boana pulchella). The results demonstrated that lambda-cyhalothrin concentration and toxicity in water were significantly lower in the Azolla spp. treatment. Furthermore, the half-life of lambda-cyhalothrin decreased from 1.2days in the unvegetated treatment to 0.4days in the vegetated treatment. The vegetated treatment also resulted in a significantly lower mortality rate for both H. curvispina and C. decemmaculatus. However, no mortality was observed in B. pulchella for any of the treatments. Sublethal effects were observed in this organism, such as lateral bending of the tail and impairment of the ability to swim, which were attenuated in the vegetated treatment. We conclude that Azolla spp. can effectively reduce the concentration and toxicity of lambda-cyhalothrin, suggesting its potential use in farm-scale best management practices to mitigate the effects of pesticide loads from adjacent crops.

Gluconeogenesis in frogs during cooling and dehydration exposure: new insights into tissue plasticity of the gluconeogenic pathway dependent on abiotic factors.

Anurans undergo significant physiological changes when exposed to environmental stressors such as low temperatures and humidity. Energy metabolism and substrate management play a crucial role in their survival success. Therefore, understanding the role of the gluconeogenic pathway and demonstrating its existence in amphibians is essential. In this study, we exposed the subtropical frog Boana pulchella to cooling (-2.5°C for 24 h) and dehydration conditions (40% of body water loss), followed by recovery (24 h), and assessed gluconeogenesis activity from alanine, lactate, glycerol and glutamine in the liver, muscle and kidney. We report for the first time that gluconeogenesis activity by 14C-alanine and 14C-lactate conversion to glucose occurs in the muscle tissue of frogs, and this tissue activity is influenced by environmental conditions. Against the control group, liver gluconeogenesis from 14C-lactate and 14C-glycerol was lower during cooling and recovery (P<0.01), and gluconeogenesis from 14C-glutamine in the kidneys was also lower during cooling (P<0.05). In dehydration exposure, gluconeogenesis from 14C-lactate in the liver was lower during recovery, and that from 14C-alanine in the muscle was lower during dehydration (P<0.05). Moreover, we observed that gluconeogenesis activity and substrate preference respond differently to cold and dehydration. These findings highlight tissue-specific plasticity dependent on the nature of the encountered stressor, offering valuable insights for future studies exploring this plasticity, elucidating the importance of the gluconeogenic pathway and characterizing it in anuran physiology.

Grazing for biodiversity: Assessing the effects of cattle management practices on wetlands and amphibian communities in central Argentina

Argentina is widely recognized as a key player in the production and consumption of beef, occupying a prominent position in the global context. However, there is a lack of comprehensive research on the environmental consequences of beef production, particularly concerning wetlands and aquatic biodiversity. We aim to assess the effects of cattle management practices (grazing types and stocking density) on wetland attributes and associated amphibian diversity in central Argentina. During two amphibian breeding seasons, we surveyed 117 wetland sites in paddocks with different cattle grazing types (continuous grazing, glyphosate-promoted pastures and rotational grazing) and stocking densities (low and high). We analyzed the effects of cattle management practices on wetland attributes, including water quality parameters, nutrient concentrations, wetland morphometry, and vegetation cover. Effects on amphibian communities were explored using richness, abundance, and species occurrence. General Mixed-effects Models revealed significant increases in nutrients (total solid dissolved, total nitrogen and soluble reactive phosphorous) and a significant decrease in vegetation cover in sites with high cattle stocking density. Likewise, we observed increased soluble reactive solids in wetlands associated with rotational grazing and decreased vegetation cover in those wetlands related to glyphosate-promoted pastures. Generalized Mixed-effects Models revealed that amphibian communities were negatively affected mainly by high stocking density, while abundance and species occurrence of hylids (Boana pulchella and Scinax squalirostris) were also affected by glyphosate-promoted pastures and rotational grazing treatments. Our results indicate that different types of grazing at low stocking density favor amphibian diversity and improve wetland water quality, therefore, reduced stocking density may result in improved wetland conditions and more diverse amphibian communities. Our results also suggest that glyphosate-promoted pastures and rotational grazing, although beneficial in terms of pasture management, may have unintended consequences on water quality in wetlands and amphibian communities. This study will contribute to our understanding of how cattle management practices influence wetland ecosystems and aquatic biodiversity in the most important cattle breeding area of South America, providing valuable insights for conservation efforts in cattle ranching landscapes.

Metabolic changes in the subtropical frog Boana pulchella during experimental cooling and recovery conditions

Frogs have developed biochemical and physiological adaptations to occupy diverse ecological niches on Earth successfully. Survival in frozen states is a fascinating strategy made possible by evolving adaptations to produce cryoprotectant solutes. The hylid frog Boana pulchella thrives in South American regions with cold climates, remaining active while enduring sporadic subzero temperatures during winter. The species' metabolic changes during subzero exposure remain unclear. Therefore, we exposed B. pulchella to cooling and recovery, assessing plasma and tissue metabolite changes. Cooling significantly reduced urea concentrations in plasma (P = 0.033), muscle (P = 0.001), heart (P = 0.009), and brain (P = 0.041) compared to acclimation. Liver glucose oxidation and glycogen synthesis were lower in cooling and recovery than in acclimation (P < 0.0001 and P = 0.0117, respectively). Muscle glycogen synthesis was lower in recovery than acclimation (P = 0.0249). These results demonstrate B. pulchella's physiological strategies during subzero exposure, likely reflecting species-specific evolutionary adaptations for brief subzero exposures that enable winter survival in its natural habitat.

Changes in call properties of Boana pulchella (Anura, Hylidae) in response to different noise conditions.

The increasing expansion of urban areas leads to the emergence of new noisy environments that can affect animal communication. Calls play a crucial role in the mating displays of anurans, and the negative impact of anthropogenic noise-induced auditory masking has been reported in several species. We investigated the acoustic variation in 96 males (n = 971 calls) of the treefrog Boana pulchella across acoustically undisturbed sites and different noise conditions, associated with urban areas (URBAN) and roads (ROAD), in Central Argentina. We analyzed the effect of anthropogenic noise conditions on 6 temporal (call duration [CD], intercall interval, first and second note duration, internote interval, and call rate) and 3 spectral (dominant frequency of first and second note, frequency difference between them) call properties. The effects of temperature and size on acoustical variables were controlled. We observed differences in all call attributes among the noise conditions, except for intercall interval. Males exposed to URBAN and ROAD exhibited significant changes in CD, dominant frequency of the second note, and frequency difference between them. URBAN males had longest internote interval, while ROAD individuals displayed increased first and second note duration and call rates. Interestingly, ROAD males exhibited immediate changes in call rate in direct response to passing heavy vehicles. Our study emphasizes the impact of anthropogenic noise on the acoustic characteristics of B. pulchella calls. Understanding how animals adapt to noisy environments is crucial in mitigating the adverse effects of urbanization on their communication systems. Future investigations should explore whether the observed call adjustments are effective in avoiding or mitigating the negative consequences of anthropogenic noise on reproductive success.

Testing the Novel Weapons Hypothesis of the Argentine Ant Venom on Amphibians

The globally invasive Argentine ant (Linepithema humile) possesses a venom lethal to some amphibian species in the invaded range. To test the novel weapons hypothesis (NWH), the effects of the toxin on the cohabiting amphibian species in the ant's native range need to be investigated. The invader should benefit from the novel chemical in the invaded range, because the species are not adapted, but the venom should not be effective in the native range. We explore the venom effects on juveniles of three amphibian species with different degrees of myrmecophagy inhabiting the ant's native range: Rhinella arenarum, Odontophrynus americanus, and Boana pulchella. We exposed the amphibians to the ant venom, determined the toxic dose, and evaluated the short- (10 min to 24 h) and medium-term (14 days) effects. All amphibian species were affected by the venom independently of myrmecophagy. In addition to amphibian sensitivity, we discuss how the differential Argentine ant abundance and density in the two ranges could be the key to the susceptibility of amphibians to the venom, resulting in the possibility of NWH. Our results confirm the potential magnitude of the impact of the Argentine ant in successfully invaded areas for the conservation of already threatened amphibians.

Realistic scenarios of pesticide exposure alters multiple biomarkers in BOANA PULCHELLA (ANURA) Adult Frogs

Imazethapyr, a post-emergent herbicide used in worldwide soybean and corn crops, induces genetic and biochemical alterations in aquatic vertebrates. This study examined the relationship between biomarkers at different organization levels and imazethapyr real-life route exposure in Boana pulchella adults. Frogs were exposed to imazethapyr-based formulation Pivot® H (10.59%) at concentrations representing possible acute routes: field runoff (S1:10 mg.L-1), exposure after direct foliar application (S2:100 mg.L-1) and during direct foliar application (S3:1000 mg.L-1). Post-exposure, endpoints levels were evaluated: organism alterations, biochemical activities and cytogenetic assays. Forty-eight hours post-exposure, antioxidant enzymes decrease, micronuclei induction and DNA damage were observed in all scenarios, while cholinesterase activity increase and body condition reduction were observed in frog-exposed to S3. Ninety-six hours post-exposure, frogs showed glutathione-S-transferase inhibition in S1, micronuclei induction in S2 and S3, and DNA-damage increase in S3. Herbicides routes of exposures in real-life could indicate that authorized applications have a risk to amphibian populations.

Genotoxic risk assessment of solar UV radiation in tadpoles from Brazilian wetlands

Solar ultraviolet (UV) radiation is an environmental genotoxic factor linked to amphibian decline. Here we assessed the genotoxic risk of UVB and UVA exposure for tadpoles from open ponds in southern Brazil, a mid-latitude region influenced by stratospheric ozone depletion. Daily UV doses were measured on the surface of a pond in Taim Ecological Station (TAIM; 32°49’24’’S; 52°38’31’’W) on a cloudless summer day to predict the worst-case scenario for UV-induced DNA damage. Pond descriptors were related to the use of microhabitats by Boana pulchella tadpoles in two ponds over the climate seasons of 2013 and 2014. Our results indicate that shaded microhabitats were more frequent than unshaded ones in autumn, winter, and spring but not in summer. Hence, the penetration of UV radiation into the water of unshaded microhabitats was evaluated through laboratory experiments with artificial UV sources and pond water samples. Physical and biological sensors were applied in the experiments to measure the incident UV radiation and its genotoxic action. By integrating field and laboratory data, we demonstrate that low doses of biologically effective UV radiation reached the tadpoles in autumn, winter, spring, and early summer due to a high proportion of shaded microhabitats and a high concentration of solids in unshaded microhabitats. However, the relative reduction of shaded microhabitats jointly with a declining water level in late summer may have exposed tadpoles to high UV doses. Our experiments also indicate that solar UVB radiation, but not UVA, is primarily responsible for the induction of DNA pyrimidine dimers in organisms living under the surface of aquatic ecosystems. The present work highlights the determinant role of wetland descriptors for minimizing the genotoxic potential of UV radiation and its consequences for amphibians.

Structure and function of cationic hylin bioactive peptides from the tree frog Boana pulchella in interaction with lipid membranes

Amphibians have a great diversity of bioactive peptides in their skin. The cDNA prepro-peptide sequencing allowed the identification of five novel mature peptides expressed in the skin of Boana pulchella, four with similar sequences to hylin peptides having a cationic amphipathic-helical structure. Whole mature peptides and some of their fragments were chemically-synthesized and tested against Gram-positive and Gram-negative bacterial strains. The mature peptide hylin-Pul3 was the most active, with a MIC= 14 µM against Staphylococcus aureus. Circular dichroism assays indicated that peptides are mostly unstructured in buffer solutions. Still, adding large unilamellar vesicles composed of dimyristoyl phosphatidylcholine and dimyristoylphosphatidylglycerol increased the α-helix content of novel hylins. These results demonstrate the strong influence of the environment on peptide conformation and highlight its significance while addressing the pharmacology of peptides and their biological function in frogs.

Vocal repertoire and acoustic variation in a treefrog (Boana ericae) (Amphibia, Anura, Hylidae) endemic to the Chapada dos Veadeiros, Central Brazil

ABSTRACT Despite the great diversity of Brazilian anuran species, there are still many knowledge gaps about the acoustic communication of these animals. Among the species, those classified as ‘Data Deficient’ have the greatest gaps in knowledge on their behavior. This is the case of Boana ericae, a species belonging to the Boana pulchella group, endemic to the Chapada dos Veadeiros National Park region, in the northeast of the state of Goiás, Central Brazil. Three distinct call types (A, B, and C) are currently known for B. ericae, with the type A call associated with attracting reproductive partners and the type C call presenting a territorial function. We recorded the vocalizations of 10 individuals of this species and found the presence of vocalizations composing three new note types (D, E, and F). Our results indicate that air temperature and body condition influenced the acoustic parameters of the advertisement and aggressive calls of this species. We also observed that some acoustic parameters of the advertisement calls can be used for individual recognition for showing greater variation among individuals than intra-individually. Our results add new information regarding the vocal repertoire in this species, expanding their known intra-individual classification of the acoustic parameters.

Color polymorphism and variation in microhabitat selection: a defensive strategy in males of Boana pulchella (Anura: Hylidae)?

Anurans vary in body coloration and frequently exhibit color polymorphism, with selection by predation pressure favoring more cryptic animals. Spatial differentiation may also favor color polymorphism in some species, since cryptic morphotypes can use substrates that better match their coloration. Boana pulchella is a polymorphic species with green and brown morphotypes, but its polymorphism remains poorly understood. The aim of the present study was to determine the functionality of color polymorphism in B. pulchella as a defense strategy. Artificial green and brown clay models were placed in environments with green and brown background to determine if the two morphotypes of B. pulchella experience different frequencies of attacks. We expected that more conspicuous morphs are attacked more frequently. The attack rate, which suggests predation intent, on the models was 9.57%. The marks observed in the artificial models were left by both mammals and birds. The number of damaged models did not significantly differ between the different background environments. The results indicate that B. pulchella morphotypes are equally cryptic to predators in the studied environments. In conclusion, probably color polymorphism in B. pulchella is not operating as a defensive strategy, and other selective forces may be acting in the maintenance of color polymorphism in this species.

Structure and Function of Cationic Hylin Bioactive Peptides from the Tree Frog Boana Pulchella in Interaction with Lipid Membranes

Structure and Function of Cationic Hylin Bioactive Peptides from the Tree Frog Boana Pulchella in Interaction with Lipid Membranes

EVALUACIÓN PRELIMINAR DE LA DIVERSIDAD DE ANFIBIOS EN LA ALAMEDA DE SAN CARLOS (MALDONADO, URUGUAY), A TRAVÉS DE UN MONITOREO ACÚSTICO PASIVO

Biological monitoring, from species to communities, is key to biodiversity conservation management, policy development and planning. Globally, amphibians are experiencing a major diversity crisis. In the case of Uruguay, there is a record of 49 species, 33% of which have been assigned to some category of threat. The objective of this work was to contribute to the generation of information on amphibian diversity through passive acoustic monitoring in the Alameda de San Carlos. During this study, 511 recordings were obtained, with a total of 2555 minutes of recording. A total of 14 amphibian species were recorded and identified. During the 22 days of sampling, temporal variation in the activity of recorded species was notorious. Boana pulchella and Scinax squalirostris were the only ones that were active during the entire sampling, while the rest of the species were active intermittently. This study represents the first work based on passive acoustic monitoring of anuran calls in our country, providing novel information that was not available for the study area and showing great potential for teaching activities in addition to scientific ones.

Seasonal variations in the intermediate metabolism in South American tree-frog Boana pulchella

Seasonal variations in the intermediate metabolism in South American tree-frog Boana pulchella

Rational design and synthesis of modified natural peptides from Boana pulchella (anura) as acetylcholinesterase inhibitors and antioxidants.

The use of acetylcholinesterase (AChE) inhibitors, antioxidants or multitarget compounds are among the main strategies against Alzheimer's disease (AD). Between AChE inhibitors, those targeting the peripheral anionic site (PAS) are of special interest. Here, we describe the rational design and synthesis of peptide analogs of a natural PAS-targeting sequence that we recently discovered, aiming at increasing its activity against AChE. We also tested their radical scavenging and metal chelating properties. Our design strategy was based on the position-specific, computer-aided insertion of aromatic residues. The analog named as W3 showed a 30-fold higher inhibitory activity than the original sequence and an improved antioxidant activity. W3 is the most potent modified natural peptide against Electrophorus electricus AChE ever reported with an IC50 of 10.42μM (± 1.02). In addition, it showed a radical scavenging activity of 47.00% ± 3.11 at 50μM and 93.47% ± 1.53 at 400μM. Since peptides are receiving increasing interest as drugs, we propose the W3 analog as an attractive sequence for the development of new peptide-based multitarget drugs for AD. Besides, this work sheds light on the importance of the aromatic residues in the modulation of AChE activity and their effect on the radical scavenging activity of a peptide.

Prepollex diversity and evolution in Cophomantini (Anura: Hylidae: Hylinae)

Abstract Several species of Cophomantini are known to have an enlarged prepollex, commonly modified as an osseous spine. We surveyed the osteology and myology of the prepollex and associated elements of 94 of the 190 species of Cophomantini, sampling all genera, except Nesorohyla. Two distinct prepollex morphologies were found: a blade-shaped and a spine-shaped morphology. We described the observed variation in 17 discrete characters to study their evolution in the most inclusive phylogenetic hypothesis for Cophomantini. Both morphologies evolved multiple times during the evolutionary history of this clade, but the origin of the spine-shaped distal prepollex in Boana and Bokermannohyla is ambiguous. The articulation of metacarpal II with the prepollex through a medial expansion of the metacarpal proximal epiphysis is a synapomorphy for Boana. The shape of the curve of the spine, and a large post-articular process of the distal prepollex, are synapomorphies for the Boana pulchella group, the latter being homoplastic in the Bokermannohyla martinsi group. Muscle character states associated with the spine-shaped prepollex are plesiomorphic for Cophomantini. We discuss evolution, function, behaviour and sexual dimorphism related to the prepollical elements. A bony spine is associated with fights between males, but forearm hypertrophy could be more related with habitat than with territorial combat.

Body size, age and growth in males populations of Boana pulchella (Anura, Hylidae).

Age determination in amphibians is crucial to the investigation of life-history traits. In this context, we studied, for the first time, the life-history traits of Boana pulchella from a sample (63 adult males) of three populations (Las Acequias, Río Cuarto, Alejandro Roca) in central Argentina using the skeletochronological method. All adults of B. pulchella studied showed recognizable bone structures that allowed age determination. The average snout-vent length of sexually mature males was 43.39 mm. The maximum observed longevity was 5 years (5 Las Acequias, 4 Río Cuarto, 3 Alejandro Roca) and minimum age at sexual maturity was 2 years (same in the three populations), with mean of 2.96 years. Body size and age were positively correlated (except in Alejandro Roca populations). The growth patterns, estimated by the von Bertalanffy growth equation, do not show differences between populations, and the growth rate decrease after the attainment of sexual maturity. In conclusion, the determination of the individual age of the different populations of B. pulchella allowed us for the first time to establish the characteristics of the life history of the species, important for future comparisons whit others populations and future conservation biology studies.