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The Campos de Palmas Wildlife Refuge (CPWR) (Brazil) is a full protection Conservation Unit (CU) formed by private properties, where the use of land and natural resources are allowed sustainably according to the rules of the CU Management Plan. Inadequate practices of land use and occupation within this CU can affect its quality and the organisms that depend on them. Considering the above, the objective of the present study was to biomonitor different land uses and occupations within the CPWR and its surroundings, during the four seasons of a year, using the bioindicators Allium cepa (cytotoxicity and genotoxicity tests) and Eisenia fetida (leakage test). The soil samples were collected in areas of silviculture, agriculture, and native grasslands within the CU and agriculture outside the limits of the CPWR. The use by silviculture (spring) and native grassland (summer) showed cytotoxic effects for A. cepa. The use by agriculture outside the CPWR (spring) showed genotoxic effect and stimulation of mitotic cell division. For the animal bioindicator, the use by agriculture within the CPWR (winter), native grassland (autumn and spring), silviculture (autumn and spring), and agriculture outside the CU (spring) showed a toxic effect. A PCA analysis showed a correlation between the results of toxicity, cytotoxicity, and genotoxicity with the presence of macronutrients and metals in the evaluated soil samples. Possibly influenced by their soil composition characteristics (silviculture and native grassland) or the management of agricultural areas. In addition, samples from the rainy season (spring) showed higher ecotoxicity. These results show that biomonitoring through different organisms is important for evaluating environmental quality and indicate the need for the implementation of preventive measures in the CPWR to avoid the toxic, cytotoxic, and genotoxic effects found. They also ensure the integrity of this CU and the active protection of the environment and biodiversity.

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Our objective was to describe and compare the diet of endemic anurans of the Atlantic Forest, as well as to evaluate the influence of biotic factors on the trophic ecology of the species. We conducted the study in Mananciais da Serra (Piraquara-PR), a transition region between mixed and dense ombrophilous forest, between January 2019 and February 2020. In this work, we describe the diet of eight species: Aplastodiscus albosignatus, Boana semiguttata, Bokermannohyla circumdata, Ischnocnema henselii, Leptodactylus notoaktites, Proceratophrys boiei, Rhinella abei and Scythrophrys sawayae. Coleoptera were important prey in the diet of most species, except for B. circumdata and L. notoaktites. All species showed potential selection for some category of prey. We observed a correlation between the size of the individuals and the volume of consumed prey in I. henselii. In general, the niche overlap between species was low, and most species had a broad trophic niche.

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The importance of amphibian bioacoustics is widely documented in ecological, taxonomical and evolutionary studies, as calls might act as a primary mechanism of reproductive isolation. The influence of air temperature and male size on the variation of the advertisement calls in anurans is widely recognized but still insufficiently analyzed in Brazilian species. Our goals were to (1) describe the advertisement call of Aplastodiscus albosignatus (Lutz & Lutz, 1938), (2) test the influence of temperature and body size on its acoustic signals and (3) evaluate the variation of within and between-male acoustic signals. Advertisement calls of A. albosignatus consist of an unpulsed note with four harmonics. In most cases, the dominant frequency is the third harmonic but, in some calls, it was the minimum frequency (first harmonic). The average duration of calls was 0.191 s, the interval between calls was 2.08 s and the repetition rate was 33 calls per minute. On average, the minimum frequency was 550.15 Hz, the maximum frequency was 3531.70 Hz and the third harmonic was 2498.9 Hz. To evaluate the effect of air temperature, and body size on the variation of call parameters, we performed generalized linear models. The most explanatory model for spectral parameters was temperature plus body size. Concerning temporal variables, the best model that explains the variation in call duration was body size, while for the interval between calls was air temperature. The maximum frequency and the frequency of the third harmonic had little variation in the calls of both the same male and different males. Thus, these parameters were considered important in species recognition.(AU)

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The Campos de Palmas Wildlife Refuge (RVS-CP) is a full protection conservation unit (CU) formed by private properties. The present study aimed to use the bioindicators Allium cepa L. (cytotoxicity and mutagenicity tests) and Eisenia fetida (avoidance test) to assess the quality of surface water of the Chopim River within the RVS-CP area and its surroundings during the four seasons of the year. To do so, water samples were collected at five points, four inside the RVS-CP area and a fifth point outside thereof. Samples from all sampling points had cytotoxic effect on A. cepa in at least one season of the year. Such a finding may be related to inadequate management practices (without land-use control) in the areas surrounding the sampling points such as forestry, native fields, pastures, agriculture, and housing areas. As for the animal bioindicator (E. fetida), only points 1 (in the winter) and 5 (in the autumn) were toxic. Concerning mutagenicity, points 1 and 4 (in the spring), 1 and 2 (in the summer), and 3 (in the autumn) showed mutagenic effect on A. cepa meristematic cells, therefore only within the RVS-CP area. Overall, these results show that biomonitoring can be an ally of the residents of the RVS-CP area in controlling management practices, aiming to bring together economic support and conservation of resources, especially water.

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Complex interactions among hosts, pathogens, and the environment affect the vulnerability of amphibians to the emergence of infectious diseases such as chytridiomycosis, caused by Batrachochytrium dendrobatidis (Bd). Boana curupi is a forest-dwelling amphibian endemic to the southern Atlantic Forest of South America, a severely fragmented region. Here, we evaluated whether abiotic factors (including air and water temperature, relative air humidity, and landscape) are correlated with chytrid infection intensity and prevalence in B. curupi. We found individuals infected with Bd in all populations sampled. Prevalence ranged from 25-86%, and the infection burden ranged from 1 to over 130000 zoospore genomic equivalents (g.e.) (mean ± SD: 4913 ± 18081 g.e.). The infection load differed among populations and was influenced by forest cover at scales of 100, 500, and 1000 m, with the highest infection rates recorded in areas with a higher proportion of forest cover. Our results suggest that the fungus is widely distributed in the populations of B. curupi in southern Brazil. Population and disease monitoring are necessary to better understand the relationships between host, pathogen, and environment, especially when, as in the case of B. curupi, threatened species are involved.

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Animals have a long history of assessing ecosystem responses to environmental disturbances, and amphibians stand out for presenting themselves as good animal model and bioindicators of environmental quality. The main purpose of the present work was to investigate the cellular effects of contamination of waters of the Marrecas River, located in the southwest of the state of Paraná, Brazil. Therefore, the objective of the present study was to investigate and monitor the cytotoxic and mutagenic effect, with bullfrog tadpoles, and to discuss these effects with land use along this hydrographic basin. Mutagenic effects were determined by micronucleus assay, and cytotoxicity by other nuclear changes, such as segmented cells, binucleated cells, cells with buds and reniform cells. Water samples were obtained at nine sites along the Marrecas River, covering areas with rural and urban hydrological contribution. For each site, four samples were collected, along the years 2017 and 2018, encompassing the four seasons (summer, autumn, winter, and spring). The results showed mutagenic and cytotoxic effect in four sampling sites, and only cytotoxic effect in other four sites. These effects may be due, possibly, to the use of different agrochemicals across the hydrographic basin region, which have predominant hydrological contributions from crops. Data of this study indicate the presence of cytotoxic and mutagenic contaminants in the waters of the Marrecas River, which can generate environmental problems on the river fauna/flora, and can also affect the local population health.

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Data on geographic distribution and ecology are essential when defining the conservation status of aspecies. Herein, we present new information about the geographic distribution, habitat and advertisement callof Ischnocnema henselii, an endemic species of the southern Atlantic Forest. We conducted this study in forestfragments in the state of Santa Catarina, Southern Brazil, from January 2008 to November 2012. We recordedI. henselii in eight fragments over 100 ha in size, both inside and outside conservation units. We often observedless than 20 males in calling activity. The calling activity was daytime and nighttime, and more intense in therain. We observed individuals perched on vegetation, on leaves or branches, on the leaf litter or under treetrunks. Duration of advertisement calls ranged from 14.71 ± 1.68 s (n = 11 calls) and interval between noteswas 0.22 ± 0.13 s (n = 99 notes). We observed that the advertisement calls had a feature that we call “alternatingmodulation” in the amplitude of the notes. We suggest that further bioacoustic studies should analyze therelevance of this alternating modulation, to understand its role in communication. Although I. henselii is a habitatspecialist, it is currently considered a common species, classified as Least Concern by Brazilian and IUCNlists. In the region studied, characterized by small and isolated remnants of the Atlantic Forest, I. henselii is notan abundant species and its spatial distribution is always associated with wet forest fragments. The increasingdegradation of forest remnants in this region causes microclimatic and structural changes that may compromisethe permanence of strictly forest and moisture-dependent species for reproduction, such as I. henselii.

Informações sobre distribuição geográfica e ecologiasão essenciais para a definição do estado de conservação de uma espécie. Aqui, nós apresentamos novasinformações sobre a distribuição geográfica, habitat e canto de anúncio de I. henselii, um anfíbio endêmicodo Sul da Mata Atlântica. Nós conduzimos o estudo em fragmentos florestais no estado de Santa Catarina, sul do Brasil, de janeiro de 2008 a novembro de 2012. Registramos I. henselii em oito fragmentos com tamanhoacima de 100 ha, no interior ou fora de unidades de conservação. Nós frequentemente observamos menosde 20 machos em atividade de vocalização. As atividades de vocalização foram registradas de dia e à noite eeram mais intensas sob chuva. Observamos indivíduos empoleirados na vegetação, sobre folhas ou galhos, nofolhiço ou sob troncos de árvores caídas. A duração do canto de anúncio foi 14,71s ± 1,68 (n = 11 cantos) e ointervalo entre as notas foi de 0,22s ± 0,13 (n = 99 notas). Os cantos de anúncio apresentaram o que chamamosde “modulação alternada” na amplitude das notas. Sugerimos que outros estudos analisem a relevânciadesta “modulação alternada”, visando compreender a sua relação com a comunicação nesta espécie. EmboraI. henselli seja especialista de habitat, é considerada comum, classificada na categoria “preocupação menor”nas listas brasileira e mundial de espécies ameaçadas de extinção. Na região de estudo, caracterizada porremanescentes pequenos e isolados de Mata Atlântica, I. henselii é pouco abundante e encontrada somenteem fragmentos florestais úmidos. A crescente degradação dos remanescentes florestais nesta região ocasionamudanças microclimáticas e estruturais que podem comprometer a permanência de espécies estritamenteflorestais e dependentes da umidade para reprodução, como I. henselii.

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Data on geographic distribution and ecology are essential when defining the conservation status of aspecies. Herein, we present new information about the geographic distribution, habitat and advertisement callof Ischnocnema henselii, an endemic species of the southern Atlantic Forest. We conducted this study in forestfragments in the state of Santa Catarina, Southern Brazil, from January 2008 to November 2012. We recordedI. henselii in eight fragments over 100 ha in size, both inside and outside conservation units. We often observedless than 20 males in calling activity. The calling activity was daytime and nighttime, and more intense in therain. We observed individuals perched on vegetation, on leaves or branches, on the leaf litter or under treetrunks. Duration of advertisement calls ranged from 14.71 ± 1.68 s (n = 11 calls) and interval between noteswas 0.22 ± 0.13 s (n = 99 notes). We observed that the advertisement calls had a feature that we call “alternatingmodulation” in the amplitude of the notes. We suggest that further bioacoustic studies should analyze therelevance of this alternating modulation, to understand its role in communication. Although I. henselii is a habitatspecialist, it is currently considered a common species, classified as Least Concern by Brazilian and IUCNlists. In the region studied, characterized by small and isolated remnants of the Atlantic Forest, I. henselii is notan abundant species and its spatial distribution is always associated with wet forest fragments. The increasingdegradation of forest remnants in this region causes microclimatic and structural changes that may compromisethe permanence of strictly forest and moisture-dependent species for reproduction, such as I. henselii.(AU)

Informações sobre distribuição geográfica e ecologiasão essenciais para a definição do estado de conservação de uma espécie. Aqui, nós apresentamos novasinformações sobre a distribuição geográfica, habitat e canto de anúncio de I. henselii, um anfíbio endêmicodo Sul da Mata Atlântica. Nós conduzimos o estudo em fragmentos florestais no estado de Santa Catarina, sul do Brasil, de janeiro de 2008 a novembro de 2012. Registramos I. henselii em oito fragmentos com tamanhoacima de 100 ha, no interior ou fora de unidades de conservação. Nós frequentemente observamos menosde 20 machos em atividade de vocalização. As atividades de vocalização foram registradas de dia e à noite eeram mais intensas sob chuva. Observamos indivíduos empoleirados na vegetação, sobre folhas ou galhos, nofolhiço ou sob troncos de árvores caídas. A duração do canto de anúncio foi 14,71s ± 1,68 (n = 11 cantos) e ointervalo entre as notas foi de 0,22s ± 0,13 (n = 99 notas). Os cantos de anúncio apresentaram o que chamamosde “modulação alternada” na amplitude das notas. Sugerimos que outros estudos analisem a relevânciadesta “modulação alternada”, visando compreender a sua relação com a comunicação nesta espécie. EmboraI. henselli seja especialista de habitat, é considerada comum, classificada na categoria “preocupação menor”nas listas brasileira e mundial de espécies ameaçadas de extinção. Na região de estudo, caracterizada porremanescentes pequenos e isolados de Mata Atlântica, I. henselii é pouco abundante e encontrada somenteem fragmentos florestais úmidos. A crescente degradação dos remanescentes florestais nesta região ocasionamudanças microclimáticas e estruturais que podem comprometer a permanência de espécies estritamenteflorestais e dependentes da umidade para reprodução, como I. henselii.(AU)

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Phenotypic traits, such as the frog advertisement call, are generally correlated with interspecific genetic variation, and, as a consequence of strong sexual selection, these behaviors may carry a phylogenetic signal. However, variation in acoustic traits is not always correlated with genetic differences between populations (intraspecific variation); phenotypic plasticity and environmental variables may explain part of such variation. For example, local processes can affect acoustic properties in different lineages due to differences in physical structure, climatic conditions, and biotic interactions, particularly when populations are isolated. However, acoustic traits can be used to test phylogenetic hypotheses. We analyzed the advertisement calls of Dendropsophus elegans males from 18 sites and compared them with those of four closely related congeneric species, in order to test for differences between inter and intraspecific variation. We analyzed 451 calls of 45 males of these five species. Because males from distant sites were grouped together without population congruence, differences found in advertisement calls among individuals were not correlated with phylogeographical clades. Phylogenetic and cluster analyses of the D. elegans clades and those of closely related species grouped all five species into the same topology, as reported by previous molecular and morphological phylogenies. However, the topology of the D. elegans phylogeographical clades did not match the topology previously reported. Acoustic communication in D. elegans seems to be conserved among populations, and the phylogeographical history of the species does not explain the variation among lineages in call properties, despite some congruent phylogenetic signals evident at the species level. Based on molecular clocks retrieved from the literature, it seems that more than 6.5 million years of divergence (late Miocene) are necessary to allow significant changes to occur in the acoustic properties of these treefrog calls, making it possible to recover their phylogenetic history only based on acoustic evidence.

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Species distributed across vast continental areas and across major biomes provide unique model systems for studies of biotic diversification, yet also constitute daunting financial, logistic and political challenges for data collection across such regions. The tree frog Dendropsophus minutus (Anura: Hylidae) is a nominal species, continentally distributed in South America, that may represent a complex of multiple species, each with a more limited distribution. To understand the spatial pattern of molecular diversity throughout the range of this species complex, we obtained DNA sequence data from two mitochondrial genes, cytochrome oxidase I (COI) and the 16S rhibosomal gene (16S) for 407 samples of D. minutus and closely related species distributed across eleven countries, effectively comprising the entire range of the group. We performed phylogenetic and spatially explicit phylogeographic analyses to assess the genetic structure of lineages and infer ancestral areas. We found 43 statistically supported, deep mitochondrial lineages, several of which may represent currently unrecognized distinct species. One major clade, containing 25 divergent lineages, includes samples from the type locality of D. minutus. We defined that clade as the D. minutus complex. The remaining lineages together with the D. minutus complex constitute the D. minutus species group. Historical analyses support an Amazonian origin for the D. minutus species group with a subsequent dispersal to eastern Brazil where the D. minutus complex originated. According to our dataset, a total of eight mtDNA lineages have ranges >100,000 km2. One of them occupies an area of almost one million km2 encompassing multiple biomes. Our results, at a spatial scale and resolution unprecedented for a Neotropical vertebrate, confirm that widespread amphibian species occur in lowland South America, yet at the same time a large proportion of cryptic diversity still remains to be discovered.

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Estudos envolvendo análises bioacústicas têm possibilitado a identificação de espécies pertencentes a grupos complexos, bem como a descrição e diferenciação de espécies crípticas. Recorrendo a esta ferramenta e com o objetivo de aumentar o conhecimento sobre a composição de espécies de anfíbios em áreas de Floresta com Araucária, foram amostradas 11 áreas nos estados do Paraná e Santa Catarina. Os resultados ampliaram significativamente o conhecimento da distribuição geográfica de oito espécies de anfíbios anuros: Ischnocnema henselii, Dendropsophus anceps, atualmente classificada como criticamente ameaçada no Estado do Paraná, D. nahdereri, Scinax granulatus, Trachycephalus dibernardoi, Pseudis cardosoi, Leptodactylus araucaria e L. nanus. Alguns destes registros representam correções em identificações anteriores: Ischnocnema henselii é uma espécie críptica e algumas populações até o momento eram confundidas no Paraná com I. guentheri; Trachycephalus dibernardoi era identificada como T. imitatrix e Leptodactylus nanus como L. marmoratus. A caracterização e descrição dos cantos destas espécies também são apresentadas.

Studies involving bioacoustics analysis turned possible the identification of complex species groups, as well as the description and differentiation of cryptic species. In order to increase the knowledge on the species composition of amphibians in Araucaria Forests, we sampled eleven areas in the states of Paraná and Santa Catarina, which significantly increased the geographic distribution range of eight amphibian species: Ischnocnema henselii, Dendropsophus anceps, a currently classified as critically endangered in Paraná State, D. nahdereri, Scinax granulatus, Trachycephalus dibernardoi, Pseudis cardosoi, Leptodactylus araucaria and L. nanus. Some of the new records represent corrections of previous identifications for example: Ischnocnema henselii is a cryptic species with some populations in Paraná that have been previously mistaken to I. guentheri; Trachycephalus dibernardoi was mistaken for T. imitatrix and Leptodactylus nanus to L. marmoratus. We also characterize and describe the calls of all these species.