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Changes in the natural landscape and the indiscriminate use of pesticides can have a major impact on aquatic environments and have contributed to the worldwide decline of amphibian populations. In the present study, we sampled tadpoles of three anuran amphibians (Boana albopunctata, Physalaemus cuvieri, and Dendropsophus minutus) from ponds in six different agricultural landscapes of the Brazilian Cerrado savanna and evaluated whether and to what extent genotoxic and mutagenic damage was related to land use (the amount of forest and agricultural remnants, and related physicochemical factors) and the presence of pesticides in the water of the study ponds. We also evaluated the hepatotoxicity in P. cuvieri, which was the most abundant species at five of the six sampling points. Clomazone and atrazine were the most common pesticides found in the ponds. The B. albopunctata and P. cuvieri tadpoles presented similar patterns of DNA damage among the sampling points. The least DNA damage was found in the D. minutus tadpoles, although this species was present in only one of the study ponds. More binucleated and anucleated cells were observed in B. albopunctata, but there was no significant variation among species in terms of the number of micronuclei or other erythrocytic nuclear abnormalities. Land use and physicochemical factors did not explain the variation in the DNA damage observed in the three anurans. The hepatotoxicity analyses of P. cuvieri revealed the presence of a series of alterations, including the enlargement of the sinusoids, vacuolization of the hepatocytes, the infiltration of inflammatory cells, hepatic steatosis, and dilation of the blood vessels. The interaction between physicochemical factors and the biomarkers analyzed in the present study is complex. In particular, it will be important to better elucidate which factors are contributing, either directly or indirectly, to the decline of anuran amphibian populations, especially in threatened biomes, such as the Brazilian Cerrado. In this case, we would encourage further in situ studies that assess the ecotoxicology of the landscape, together with the systematic monitoring of aquatic environments, to guarantee the long-term integrity of amphibian populations, and those of other organisms that play an essential functional role in the ecosystem. Environ Toxicol Chem 2023;42:2422-2439. © 2023 SETAC.

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The ecotoxicological risk to vertebrates posed by zinc oxide nanoparticles (ZnO NPs) is still poorly understood, especially in animals with a biphasic life cycle, which have aquatic and terrestrial phases, such as amphibians. In the present study, we investigated whether acute exposure (7 days) to ZnO NPs and zinc chloride (ZnCl2) at three environmentally relevant concentrations (0.1, 1.0, and 10 mg L-1) induces changes in the morphology, chondrocranium, and behavior of the tadpoles of Lithobates catesbeianus (Anura: Ranidae). Tadpoles exposed to both forms of Zn did not undergo any morphological or behavioral changes at the lowest concentrations (0.1 and 1.0 mg L-1). However, the animals exposed to the highest concentration (10 mg L-1) lacked oral disc structures, were smaller in size, had a longer tail, and presented changes in the position and coiling of the intestine and malformations of the chondrocranium in comparison with the control group. This indicates that ZnO NPs and ZnCl2 altered the development of the tadpoles, causing delays in their metamorphosis and even reducing individual fitness. The tadpoles exposed to both forms of Zn at 10 mg L-1 also had reduced mobility, especially in the presence of conspecifics. Based on these findings, we emphasize the importance of studying morphological, skeletal, and behavioral biomarkers to evaluate the toxic effects of metal-based nanoparticles in amphibians.

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The production and use of titanium dioxide (TiO2) nanoparticles are increasing worldwide. The release of this substance into the environment can induce toxic effects in aquatic invertebrates and vertebrates, although the exact nature of its impacts on Neotropical amphibians is still poorly understood. In this context, the present study evaluated the toxicity of TiO2 nanoparticles and their counterpart-dissolved titanium dioxide (TiO2)-in the tadpoles of Dendropsophus minutus. The biometric parameters, DNA damage, and behavioral changes were verified in tadpoles exposed to three different concentrations (0.1 mg·L-1, 1.0 mg·L-1, and 10 mg·L-1) of TiO2 nanoparticles and dissolved TiO2 for 7 days. We verified significant DNA damage in the D. minutus tadpoles exposed to both forms of Ti, in comparison with the control group. We also identified a reduction in total size, body length, and width, and the height of the musculature of the tail of the tadpoles exposed to all concentrations of both substances in comparison with the control. In the behavioral test, the tadpoles exposed to nanoparticles and dissolved TiO2 presented reduced mobility and a tendency to be less aggregated than normal. Here, the simultaneous use of multiple biomarkers was fundamental for the reliable assessment of the adverse effects of nanomaterials on anuran amphibians and the establishment of a systematic approach to the biomonitoring of aquatic ecosystems. The present study expands our understanding of the genotoxic, morphological, and behavioral effects of TiO2 nanoparticles and dissolved TiO2 on anuran amphibians, and contributes to the establishment of further research for the more systematic assessment of the environmental risk of nanomaterials.

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Abstract: Despite increasing efforts in recent years to catalog the fish diversity of Amazonian rivers, many regions are still under-sampled, and sandy beach environments are particularly poorly understood. The present study focused on a 300 km stretch of the Acre river, in the southwestern Amazon basin, where we sampled 30 sandy beaches separated by a mean interval of 10 km. We collected 15,329 fish representing 80 species, 26 families, and nine orders. The Characiformes were the most abundant order, providing 88.24% of the individuals collected, followed by the Siluriformes, with 10.03%, while the Siluriformes had the highest species richness, with 37 species (45.0%), followed by the Characiformes, with 30 (37.5%). The most abundant species were the characiforms Knodus orteguasae and Creagruto barrigai. Reliable data on a region's biota is fundamental for the evaluation of patterns of biodiversity, and the occurrence and management of threatened species. As fish are directly affected by pollutants and the degradation of aquatic environments, further research in areas that are still poorly sampled will be essential for the elaboration of effective conservation strategies.

Resumo: Apesar dos crescentes esforços para catalogar a diversidade de peixes nos rios amazônicos, muitas regiões ainda estão sub-amostradas e os ambientes de praias arenosas são particularmente pouco compreendidos. Este estudo foi realizado ao longo de um trecho de 300 km do Rio Acre, no sudoeste da bacia amazônica, onde foram amostradas 30 praias, separadas por uma distância média de 10 km. Foram coletados 15.329 peixes, representados em 80 espécies, 26 famílias e nove ordens. Characiformes foi a ordem mais abundante, representando 88,24% dos indivíduos coletados, seguidos pelos Siluriformes, com 10,03%, enquanto os Siluriformes apresentaram a maior riqueza, com 37 espécies (45,0%), seguidas pelos Characiformes, com 30 espécies (37,5%). As espécies mais abundantes foram os characiformes Knodus orteguasae e Creagruto barrigai. Dados confiáveis sobre a biota de uma região são fundamentais para a avaliação dos padrões de biodiversidade e conhecimento sobre a ocorrência e manejo de espécies ameaçadas. Como os peixes são diretamente afetados por poluentes e pela degradação dos ambientes aquáticos, mais pesquisas em áreas que ainda são pouco amostradas serão essenciais para a elaboração de estratégias eficazes de conservação.

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Anthropogenic disturbance is a major cause of the biodiversity crisis. Nevertheless, the role of anthropogenic substrate vibrations in disrupting animal behavior is poorly understood. Amphibians comprise the terrestrial vertebrates most sensitive to vibrations, and since communication is crucial to their survival and reproduction, they are a suitable model for investigating this timely subject. Playback tests were used to assess the effects of substrate vibrations produced by two sources of anthropogenic activity- road traffic and wind turbines- on the calling activity of a naïve population of terrestrial toads. In their natural habitat, a buried tactile sound transducer was used to emit simulated traffic and wind turbine vibrations, and changes in the toads' acoustic responses were analyzed by measuring parameters important for reproductive success: call rate, call duration and dominant frequency. Our results showed a significant call rate reduction by males of Alytes obstetricans in response to both seismic sources, whereas other parameters remained stable. Since females of several species prefer males with higher call rates, our results suggest that anthropogenically derived substrate-borne vibrations could reduce individual reproductive success. Our study demonstrates a clear negative effect of anthropogenic vibrations on anuran communication, and the urgent need for further investigation in this area.

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Nanomaterials (NMs) have been used in a growing number of commercial products, and their rapid expansion could lead to their release into the aquatic environments. However, there is limited knowledge about the impact of NMs in the biota, especially the amphibians. The present study revised the historical use of amphibian species as a model system for nanoecotoxicological studies and summarized the data available in the scientific literature about the genotoxic, mutagenic, histopathological, embryotoxic and reproductive effects of NMs in different groups of amphibians. The interaction, bioaccumulation, mode of action (MoA) and ecotoxicity of NMs on amphibians were also revised. The nanoecotoxicological studies were conducted with 11 amphibian species, being eight species of the order Anura and three species of the order Caudata. Xenopus laevis was the most studied species. The studies were conducted mainly with inorganic NMs (72%) compared to organic ones. The nanoecotoxicity depends on NM behavior and transformation in the environment, as well as the developmental stages of amphibians. The known effects of NMs in amphibians were mainly reported with reactive oxygen species (ROS) production, oxidative stress, and genotoxic effects. Results emphasize the need for further studies testing the ecotoxicity of different NMs, concentrations and exposure periods at environmentally relevant approaches. Furthermore, standard protocols for nanoecotoxicological tests using amphibians are required. Revised data showed that amphibians are suitable organisms to assess the environmental impact of NMs and indicated significant research gaps concerning the ecotoxicity of NMs on freshwater ecosystems and recommendations for future researches.

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Cytogenetic data can be useful for taxonomic and phylogenetic studies, as well as to provide information about chromosome evolution. Therefore, it may help design conservation priorities for some threatened species, such as anurans. Herein, we describe the karyotypes of Scinax constrictus and Ololygon centralis, native endemic species from the Brazilian Cerrado. Chromosome preparations for both species were stained with Giemsa for morphological analyses and then impregnated by the Ag-NOR method for localization of the nucleolar organizer region (NOR). Both species had 24 chromosomes, as confirmed by meiotic analyses, which showed 12 bivalents. Chromosome morphologies presented the same pattern for Scinax and Ololygon compared to species already karyotyped in both genera. The NOR was interstitially located in the long arm of pair 7 in S. constrictus, whereas in O. centralis it was found near the centromere in the long arm of pair 1, thus diverging from what is commonly found for other Ololygon species. Therefore, we provide the first description of the karyotype of O. centralis and the first report of the localization of the NOR for the karyotype of both species. Our study increases the cytogenetic knowledge in species of the genera Scinax and Ololygon, and provide support for further studies on the taxonomy, ecology, and evolution of hylid anurans.

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Studies in remnants of semi-deciduous seasonal forest of the Cerrado are needed to fill sampling gaps and improve basic knowledge of biodiversity. This study presents data on the herpetofauna of Parque Estadual Altamiro de Moura Pacheco, one of the last protected areas of seasonal forest in the core region of the Brazilian Cerrado. Fourteen sites were sampled between March 2007 and April 2008 using pitfall traps and active searches. A total of 35 anuran species were recorded, most of which were found in hygrophilous environments near forest fragments, open areas and riparian or gallery forest. A total of 29 reptile species were recorded, most of which were found in semi-deciduous forest and riparian or gallery forest. Fourteen of the amphibians and five of the reptiles are endemic to the Cerrado. The richness of amphibians and reptiles found in Parque Estadual Altamiro de Moura Pacheco is regionally representative, with communities typical of forest environments.(AU)

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Dendropsophus cruzi (Pombal Bastos, 1998) is a small hylid (male snout-to-vent length = 16.3-19.4 mm; female SVL = 21.3-25.0 mm) that is allocated in the D. microcephalus group (Faivovich et al. 2005). It is commonly found in Open and forested areas from Central Brazil to Provincia Velasco, Departamento de Santa Cruz, Bolivia (Frost 2017; Tessarolo et al. 2016). Pombal Bastos (1998) described the advertisement call of D. cruzi as a single pulsed note. Posteriorly, studies uncovered relationships between dominant frequency variation in the advertisement calls of D. cruzi and variation in the SVL and mass of males (Bastos et al. 2003), as well as the existence of a clinal geographic pattern in the variation of acoustic parameters of these calls (Tessarolo et al. 2016). However, the acoustic communication in anurans is usually mediated by more than one vocalization type (Toledo et al. 2015). Due the importance of call types in the social context of anurans (e.g., Reichert 2011; Forti et al. 2017), we expand the vocal repertoire of D. cruzi by describing its aggressive call.

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