Amphibian Diversity: Where everything starts to flood, Cáceres Municipality, North Pantanal, Central­West Brazil

Faunal inventories contribute to our understanding of regional diversity, and are fundamental for policy and de-cision-making regarding the management and conservation of large natural areas. This study aimed to inventory and com-pile information on amphibian species occurring in the North Pantanal region, in the municipal limits of Cáceres, Mato Grosso, Brazil. We used three methods to inventory amphibian species: (1) fieldwork, (2) analysis of specimens deposited in scientif-ic collections, and (3) literature reviews. We registered 49 amphibian species in Cáceres. Of them, 48 species belonged to the Anura order and were distributed across eight families and 20 genera, and one species belonged to the Gymnophiona order (Siphonops paulensis). The families Leptodactylidae (20 spp.) and Hylidae (17 spp.) were dominant in terms of richness, ac-counting for 75.5% of all species found in Cáceres. The remaining families had between four and one species each. The mu-nicipality is strongly influenced by non-forested formations (e.g., the Cerrado and Pantanal) and presents a high species rich-ness for a non-forested location in Brazil. Our findings highlight Cáceres as one of the richest areas in amphibian species in the North Pantanal region, expanding our knowledge regarding frog diversity. This study provides a foundation for future conser-vation strategies and additional assessments of amphibian species in light of potential population declines and other emerg-ing threats.(AU)

Hotspots and hotmoments of wildlife roadkills along a main highway in a high biodiversity area in Brazilian Amazonia / Hotspots e hotmoments de atropelamentos de animais silvestres ao longo de uma rodovia em uma área de alta biodiversidade na Amazônia brasileira

Wildlife roadkills have become a concern in the Amazon biome due to the opening of major roads in recent decades. In this study, we aimed to describe wildlife roadkills in a 100-km stretch of the BR-163 highway, in western Pará state, determining which vertebrate groups are most affected and whether there are spatial (hotspots) and temporal (hotmoments) aggregations of roadkills. From July 2019 to June 2020, we carried out 25 surveys at 15-day intervals, from a vehicle at a maximum speed of 40 km h-1. We recorded 351 individuals at an observed rate of 0.14 ind km-1 day-1. Despite their lower detectability and faster carcass removal rate from the road due to small size, most recorded roadkills were amphibians (0.066 ind km-1 day-1). We mapped several hotspots along the study stretch considering the total number of animals recorded, and separately for amphibians and reptiles. Multiple linear regression analyses indicated that the number of roadkills of all vertebrates, amphibians and reptiles recorded are influenced by temperature and precipitation. Information on places with the highest incidence of roadkills can support actions such as the installation of underpasses and fences, aimed at reducing the impacts on wild vertebrates of this Amazonian highway.(AU)

Atropelamentos de fauna silvestre tornaram-se preocupantes no bioma amazônico devido à abertura de grandes rodovias nas últimas décadas. Neste estudo objetivamos caracterizar a fauna silvestre atropelada em um trecho de 100 km da rodovia BR-163, no oeste do estado do Pará, determinando quais grupos de vertebrados são mais afetados e se há agregações espaciais (hotspots) e temporais (hotmoments) de atropelamentos. De julho 2019 a junho 2020 realizamos 25 amostragens a intervalos de 15 dias, a partir de um veículo a uma velocidade máxima de 40 km h-1. Registramos 351 indivíduos a uma taxa observada de 0.14 ind km-1 dia-1. Apesar de sua menor detectabilidade e taxa de remoção mais acelerada da rodovia devido ao seu pequeno porte, a maioria dos atropelamentos foi de anfíbios (0.066 ind km-1 dia-1). Mapeamos diversos hotspots ao longo do trecho estudado considerando o número total de animais registrados, e, separadamente, para anfíbios e répteis. Análise de regressão linear múltipla indicou que o número total de registros de vertebrados atropelados, e o de anfíbios e répteis é influenciado por temperatura e precipitação. Informação sobre locais com maior incidência de atropelamentos pode subsidiar ações como a instalação de estruturas de passagem de fauna e cercas de proteção, visando a diminuição dos impactos sobre vertebrados silvestres desta rodovia amazônica.(AU)

Characterization of nonpathological intrascleral cartilage in the domestic sheep (Ovis aries) / Caracterização de cartilagem intraescleral não patológica em ovinos (Ovis aries)

ABSTRACT: Birds, cartilaginous and teleost fish, reptiles, and some amphibians have intrascleral cartilage and/or bone; however, these are rarely reported in therian mammals. This study aimed to investigate and characterize a nonpathological formation of cartilage in the posterior sclera of sheep macroscopically, histologically, and by immunohistochemical exam (IHC). Ninety eyes from 45 domestic sheep were collected, underwent gross examination, fixed in formalin, and embedded in paraffin for the microscopical assessment. Sections with histological shreds of cartilage were selected to perform IHC to confirm the presence of cartilage. Intrascleral cartilage was detected in 60 eyeballs (66.66%) from 37 sheep (82.22%). A slight whitish thickening was grossly seen in the posterior sclera. The histologic exam revealed a few scattered, isolated chondrocytes to larger aggregates of cartilaginous islands in the posterior sclera. Eighteen (30%) of 60 eyeballs revealed marked anti-collagen type II immunolabeling. The development of cartilaginous structures in the eyes is considered rare in mammalian animals. The high occurrence of intrascleral cartilage in the examined sheep eyes suggests that this finding corresponds to an anatomical component of sheep sclera, despite the age, breed, or body condition.

RESUMO: Aves, peixes cartilaginosos e teleósteos e répteis, bem como alguns anfíbios possuem cartilagem e/ou osso intraescleral, contudo, isso é raramente descrito em marsupiais. O objetivo deste trabalho foi investigar e caracterizar a formação não patológica de cartilagem hialina na esclera posterior de ovinos, por exame macroscópico, histológico e imuno-histoquímico. Olhos de 45 ovinos foram coletados, submetidos a exame macroscópico, fixados em formalina a 10% tamponada e incluídos em parafina para o exame histológico. Amostras com evidência de formação cartilaginosa ao exame histológico foram selecionadas para a realização da técnica de imuno-histoquímica para confirmação. Cartilagem intraescleral foi detectada em 60 amostras oculares (66,66%) de 37 ovinos (82,22%), caracterizada na avaliação macroscópica como discretos espessamentos esbranquiçados na esclera posterior. O exame histológico revelou formações que variaram de condrócitos isolados e dispersos a grandes agregados, formando ilhas cartilaginosas na esclera. Dezoito (30%) de 60 bulbos oculares mostraram imunomarcação anti-colágeno tipo II. O desenvolvimento de estruturas cartilaginosas nos bulbos oculares é considerado raro em mamíferos. A identificação de cartilagem intraescleral em 66% dos olhos ovinos avaliados no estudo indica que a formação de cartilagem corresponde a um componente anatômico normal na esclera de ovinos, independente de idade, raça ou condição corporal.

Biobanks, offspring fitness and the influence of developmental plasticity in conservation biology

Mitigation of the widely known threats to the world's biodiversity is difficult, despite the strategies and actions proposed by international agreements such as the United Nations Framework Convention on Climate Change (UNFCCC) and the Convention on Biological Diversity (CBD). Nevertheless, many scientists devote their time and effort to finding and implementing various solutions to the problem. One potential way forward that is gaining popularity involves the establishment of biobank programs aimed at preserving and storing germplasm from threatened species, and then using it to support the future viability and health of threatened populations. This involves developing and using assisted reproductive technologies to achieve their goals. Despite considerable advances in the effectiveness of reproductive technologies, differences between the reproductive behavior and physiology of widely differing taxonomic groups mean that this approach cannot be applied with equal success to many species. Moreover, evidence that epigenetic influences and developmental plasticity, whereby it is now understood that embryonic development, and subsequent health in later life, can be affected by peri-conceptional environmental conditions, is raising the possibility that cryopreservation methods themselves may have to be reviewed and revised when planning the biobanks. Here, I describe the benefits and problems associated with germplasm biobanking across various species, but also offer some realistic assessments of current progress and applications.(AU)

Vertebrates' roadkill in the southern region of the Atlantic Forest, Paraná coast - Brazil / Padrões de atropelamentos de vertebrados na região sul da Mata Atlântica, litoral do Paraná - Brasil

Being a significant global biodiversity hotspot, the Atlantic Forest has been drastically reduced by human activities. Among the anthropic activities that most affect the biodiversity of this biome is the construction and operation of roads and highways. Between harmful effects of these infrastructures, wildlife roadkill is currently considered one of the biggest causes of mortality of wild vertebrates. This study evaluated patterns of vertebrates' roadkill on two roads in the coastal region of the largest continuous remnant of the Brazilian Atlantic Forest. For twelve months, we carry out weekly samples with a motor vehicle at a constant speed of 40 km/h to search for carcasses along the roads. All carcasses found were georeferenced and identified to the lowest possible taxonomic level. Then, using Siriema v.2.0 software, we analyzed roadkill aggregation and the spatial distribution of hotspots of wildlife roadkill. In 43 sampling days, 209 road-killed animals were registered (average roadkill rate of 0.105 and 0.111 animals/kilometer/day for PR-407 e PR-508, respectively). Extrapolating the rates found, we estimate that about 1,773 animals can be roadkill every year on these roads. The most affected groups were birds (33.01%) and amphibians (30.62%), followed by reptiles (19.13%) and mammals (17.33%). Warmer months had the highest roadkill rates. We found two critical roadkill hotspots for the PR-407 (kilometer 11.7 to 12.5 and kilometer 14.7 to 16.7). For the PR-508, we found a critical point of 5.2 km (kilometer 5 to 10.2). As a short-term measure, we recommend installing speed reducers in the identified stretches and implementing environmental education campaigns with residents and tourists, especially during the summer months, aiming to mitigate the roadkill on both roads. However, due to the importance and environmental fragility of the area, we emphasize the need for running periodic road ecology and local wildlife population viability studies in the medium and long term.

Sendo um importante hotspot de biodiversidade global, a Mata Atlântica tem sido drasticamente reduzida pelas atividades humanas. Entre as atividades antrópicas que mais afetam a biodiversidade desse ecossistema está a construção e operação de estradas e rodovias. Considerado um dos efeitos mais nocivos desses empreendimentos, o atropelamento de animais silvestres é atualmente uma das maiores causas de mortalidade de vertebrados silvestres. Neste estudo, avaliamos os padrões de atropelamentos de pequenos vertebrados em duas estradas na região litorânea do maior remanescente contínuo de Mata Atlântica brasileira. Durante doze meses realizamos coletas semanais com um veículo motorizado a uma velocidade constante de 40 km/h para busca de carcaças ao longo das estradas. Todas as carcaças encontradas foram georreferenciadas e identificadas até o menor nível taxonômico possível. Em seguida, usando o software Siriema v.2.0, analisamos a distribuição e os padrões espaciais dos eventos para identificar pontos de atropelamento de animais selvagens. Em um total de 43 dias de amostragem, foram registrados 209 animais atropelados (taxa média de atropelamentos de 0,105 e 0,111 animais/quilômetro/dia para PR-407 e PR-508 respectivamente). Extrapolando os índices encontrados, podemos estimar que cerca de 1.773 animais podem ser atropelados todos os anos nessas estradas. Os grupos mais afetados foram aves (33,01%) e anfíbios (30,62%), seguidos de répteis (19,13%) e mamíferos (17,33%). Os meses mais quentes tiveram as maiores taxas de atropelamentos. Durante o período encontramos dois pontos críticos de atropelamentos para a rodovia PR-407 (quilômetro 11,7 a 12,5 e quilômetro 14,7 a 16,7). Para a rodovia PR-508, encontramos um ponto crítico de 5,2 km (quilômetro 5 a 10,2). Como medida de curto prazo, recomendamos a instalação de redutores de velocidade nos trechos identificados e a implementação de campanhas de educação ambiental com moradores e turistas, principalmente nos meses de verão, visando mitigar o atropelamento em ambas as vias. No entanto, devido à importância e fragilidade ambiental da área, ressaltamos a necessidade de estudos de ecologia viária e de viabilidade populacional da fauna local a médio e longo prazo.

Bem-estar dos organismos aquáticos e protocolos de avaliação para aquicultura / Welfare of aquatic organisms and assessment protocols for aquaculture

Peixes, anfíbios, répteis, moluscos cefalópodes e crustáceos decápodes são sencientes, e por isso, procedimentos de cultivo e de abate desses animais precisam atender aos critérios de bem-estar. Protocolos de bem-estar têm sido propostos para espécies da aquicultura de relevância econômica trançando novos rumos para atividade, mas infelizmente na maioria das propriedades brasileiras, não são adotados. Nesta revisão, contém informações sobre o cenário atual e os métodos de avaliação de bem-estar dos organismos aquáticos disponíveis na literatura científica, a fim de contribuir para a implantação de métodos, processos e estruturas que garantam o bom estado fisiológico e comportamental durante a vida e morte dos animais em cativeiro. Os dados disponíveis indicam que o estresse, a sanidade, as condições nutricionais e os métodos de insensibilização são os principais indicadores de bem-estar na aquicultura. Portanto, é imprescindível garantir estruturas adequadas ao sistema de produção, protocolos de vacinação, dietas balanceadas compostas principalmente por fontes protéicas de qualidade, depuração pré-abate e métodos de insensibilização que levam à perda de consciência imediata.

Fish, amphibians, reptiles, cephalopod molluscs and decapod crustaceans are sentient, so these animals' cultivation and slaughter procedures must meet welfare criteria. Welfare protocols have been proposed for aquaculture species of economic relevance, tracing new directions for the activity, but unfortunately, in most Brazilian properties, they are not adopted. This review contains information about the current scenario and the methods of evaluating the welfare of aquatic organisms available in the scientific literature to contribute to the implementation of methods, processes and structures that ensure a good physiological and behavioral state during the life and death of the animals in captivity. Available data indicate that stress, health, nutritional conditions and stunning methods are the main indicators of welfare in aquaculture. Therefore, ensuring adequate structures for the production system, vaccination protocols, balanced diets composed mainly of quality protein sources, pre-slaughter purification, and stunning methods that lead to immediate loss of consciousness is essential.

Amphibians and reptiles from a protected area in western Brazilian Amazonia (Reserva Extrativista do Baixo Juruá)

Sampling gaps across the logistically challenging and extremely biodiverse Amazonia largely hamper our understanding of broad-scale amphibian and reptile diversity patterns in this ecosystem. The Juruá River basin, a southwestern tributary of the Amazon River, is one of these undersampled areas, with only punctual information documented for these vertebrates that are spatially or temporally biased. This is especially the case for the lower-middle courses of the Juruá River, which also has comparatively less protected areas than its upper course. In order to fill some biodiversity knowledge gaps associated with amphibians and reptiles in this river basin, we combined results of our field expeditions carried out in 1992, 2005-2007, and 2018 to the Reserva Extrativista (Extractive Reserve) do Baixo Juruá, a Brazilian protected area in the right bank of the lower Juruá River. Amphibians and reptiles were sampled using four complementary methods: active surveys, pitfall traps, funnel traps, and trammel nets. We identified species or updated their taxonomic status with a reanalysis of the external morphology of the preserved material in the light of novel taxonomic literature (more than 1,500 specimens) and employment of DNA barcoding analyses for some newly collected specimens with contentious taxonomic status. Our combined sampling evidenced 149 species of amphibians and reptiles occurring in this protected area (72 amphibians, 68 squamates, six chelonians, and three crocodilians). Recorded species highlight the value of the lower Juruá River region as harboring quite diverse assemblages for these vertebrates, with species typical of the western and southwestern Amazonia sub-regions. Remarkable species records are presented, as well as accounts on species with lower taxonomic resolution. Furthermore, we discuss the biogeographic affinities of recorded assemblages based on the species geographic range and preferred habitats, and the value of this protected area to preserve the regional biological diversity.(AU)

Effects of low-intensity cattle ranching on amphibians in the Ñeembucú Wetland Complex, Paraguay / Efectos de la ganadería de baja intensidad sobre los anfibios en el Complejo de Humedales Ñeembucú, Paraguay

Cattle in wetlands impact water quality through waste excretion, which deposits excess nutrients, as well as decreasing the biomass and height of vegetation through trampling and herbivory. Amphibians are sensitive to these changes due to their porous skin and reliance on vegetated microhabitats. Previous studies examining the effect of cattle on amphibians in wetlands report conflicting results, exemplifying the need to avoid overgeneralizations and instead examine specific environments. In the Ñeembucú Wetland Complex in Paraguay, low-intensity cattle ranching is a common practice. This study seeks to understand how the presence of cattle in these operations impacts water chemistry and amphibian species richness, as well as determining which cattle effects (eutrophication, nutrient loading, and decreased vegetation height diversity) have the greatest impact on amphibian communities. We anticipated that increased cattle presence would negatively impact wetland condition, and consequentially lower amphibian species richness. Data was collected over seven weeks at Estancia Santa Ana, a low-intensity cattle ranch in Pilar, Paraguay. Cattle presence was measured through cow footprints and feces densities, while a vegetation survey and water quality testing (nitrate, phosphate, and dissolved oxygen) served as environmental metrics. Pitfall trapping was used to gather information on amphibian species richness and community composition. A Pearson parametric correlation test and Factor Analysis of Mixed Data in R were then used to understand the relationship between variables. Very few statistically relationships were found between variables, and those that existed showed a very weak correlation. This suggests that cow presence does not have as strong of an impact on water quality or amphibian species richness as expected. Terrestrial and aquatic vegetation were found to explain much of the variation among the data, which verifies the importance of amphibians' microhabitats. While future research comparing generalist and specialist species in the area is necessary, these initial results suggest a hopeful future for collaboration on environmental efforts with low-intensity cattle ranchers.(AU)

Efectos de la ganadería de baja intensidad sobre los anfibios en el Complejo de Humedales Ñeembucú, Paraguay. La ganadería en los humedales afecta a la calidad del agua por excreción que añade nutrientes excesivos, además de reducir la altura y biomasa de vegetación a causa de pisar y comerla. Los anfibios son sensibles a estos cambios a causa de su piel porosa y su dependencia de la vegetación como hábitat. Estudios anteriores que investigaban los efectos del ganado en anfibios descubrieron resultados en conflicto, y así es importante evitar generalizar sobre todas las especies y ambientes. En los humedales de Ñeembucú en Paraguay, los ranchos con una densidad baja de vaca y un cambio mínimo al ambiente son muy comunes. Esta investigación trata de entender como la presencia del ganado en los ranchos afecta la calidad del agua y la riqueza de especies de anfibios, además de cuales factores (eutrofización, nutrientes excesivos o menos diversidad de vegetación) tienen el impacto más grande en los anfibios. Esperábamos que una presencia más alta de ganado causaría daño a la calidad del agua, y por eso, habría menos especies de anfibios presentes. Los datos se recogieron por siete semanas en Estancia Santa Ana, una estancia cerca a Pilar, Paraguay. La presencia de la ganadería se midió por la densidad de heces y huellas, y el medioambiente se analizó por un estudio de la vegetación y pruebas del agua (nitrato, fosfato y oxígeno). Se usan trampas pozos para capturar las ranas y coleccionar información sobre la riqueza de especies y la composición de la comunidad. Se usa la prueba de correlación paramétrica de Pearson y un Factor Analysis of Mixed Data en R para entender las relaciones entre las variables. Pocas relaciones de significado estadístico existen, y las que existen muestran una correlación muy débil. Eso sugiere que la ganadería no tenga un impacto muy grande en la calidad del agua o la riqueza de las especies como esperábamos. Vegetación terrestre y acuática explica la mayoría de la variación en los datos, y eso confirma la importancia de los hábitats de los anfibios. Los resultados sugieren un futuro optimista en cuanto a la colaboración entre los esfuerzos de conservación y los rancheros.(AU)

Histopathological changes in Lithobates catesbeianustadpoles used as biomarkers of pesticide poisoning / Alterações histopatológicas utilizadas em girinos de Lithobates catesbeianuscomo biomarcadores de intoxicações causadas por pesticidas

The use of biological indicators has increased in recent years with the aim of investigating environ-mental pollution in aquatic environments that are vulnerable to the constant use of pesticides. Some biomarkers can help assess the health status, indicating physical, metabolic, and behavioral changes under acute and sublethal poisoning. The mixture of the active ingredients cyproconazole and pico-xystrobin is a widely used fungicide for the control of pests in cotton, rice, coffee, sugarcane, corn, soybean, and wheat. The objective of this study was to verify the occurrence of possible histopatholo-gical lesions in the liver and kidneys of bullfrog tadpoles (Lithobates catesbeianus) caused by a fungi-cide commercial formula composed of picoxystrobin and cyproconazole. The animals were subjected to different concentrations of the fungicide to determine the median lethal concentration (LC50-96h = 0.05 mg L-1), that is, the lethal dose for 50% of the animals in 96 h. After determining the value of LC50-96h, the animals were subjected to three sublethal concentrations (LC50-96h/2, LC50-96h/10, and LC-50-96h/100). Through histological biomarkers, it was verified that this fungicide changed the morphology of the animals' kidney and liver tissues in a chronic way, impairing the functioning of organs that are essential for their survival and metamorphosis, which can result in an imbalance in the biodiversity of aquatic ecosystems.(AU)

O uso de indicadores biológicos tem aumentado nos últimos anos, com o intuito de investigar a po-luição ambiental em ambientes aquáticos que são vulneráveis ao constante uso de pesticidas. Alguns biomarcadores podem ajudar a avaliar o estado de saúde, indicando alterações físicas, metabólicas e comportamentais de intoxicações agudas e subletais. A mistura dos ingredientes ativos picoxistrobina e ciproconazol é amplamente usada como fungicida para o controle de pragas da cultura de algodão, arroz, café, cana-de-açúcar, milho, soja e trigo. O objetivo deste trabalho foi verificar a ocorrência de possíveis lesões histopatológicas em fígado e rins de girinos de rã-touro (Lithobates catesbeianus) causadas pela mistura dos fungicidas picoxistrobina e ciproconazol. Os animais foram submetidos a diferentes concentrações do fungicida para determinação da concentração letal mediana (CL50-96h = 0,05 mg L-1), ou seja, a dose letal para 50% dos animais em 96 horas. Após a determinação do valor da CL50-96h, os animais foram submetidos a três concentrações subletais (CL50-96h/2, CL50-96h/10 e CL50-96h/100). Através dos biomarcadores histológicos, a pesquisa verificou que esse fungicida alterou a morfolo-gia dos tecidos renais e hepáticos dos animais de maneira crônica, prejudicando o funcionamento de órgãos que são fundamentais para sua sobrevivência e metamorfose, o que pode resultar em um desequilíbrio para a biodiversidade dos ecossistemas aquáticos.(AU)

A three-year Herpetofauna survey from one of the largest remnants of the Atlantic Rainforest Biome (Reserva Natural Vale)

Here we present a three-year survey of amphibian and reptile species registered in the Vale Natural Reserve (VNR), located in the north of Espírito Santo, southeastern Brazil. The VNR, along with the Sooretama Biological Reserve (SBR) and other surrounding areas, form a forest block that constitutes one of the largest remnants of continuous forest in the Atlantic Forest biome. We systematically sampled the herpetofauna community from 2015 to 2018, using the method of active search method in 27 plots of RAPELD distributed in the different types of vegetation present in the area. We recorded 39 species of amphibians belonging to the order Anura, distributed in seven families and 40 species of reptiles belonging to 20 families distributed in three orders: Crocodylia, Testudines and Squamata. The supplementation the samplings with those records from occasional encounters was important for the survey of the region's herpetofauna, mainly for reptiles. We added four species (two lizards, one amphisbaenid and one snake) to the known list for the VNR-SBR forest block: Micrablepharus maximiliani, Strobilurus torquatus, Amphisbaena alba and Tantilla sp. We demonstrated that VNR contains a high species richness of amphibians and reptiles, with a high number of anurans being endemic to the Atlantic Forest biome. The present species inventory provided the most current approximation of known the richness and composition of species of the existing herpetofauna at Vale Natural Reserve, since the period of the gathering of records is quite recent (2015-2018). Also, it brings the knowledge of some community parameters as species richness and composition not only for the whole area of the Vale Natural Reserve, but also for its different vegetation types, with no inclusion of environments outside this reserve. Furthermore, the presence of threatened species, the addition of four species to the current list, and the potential for new species to occur, reinforce the role of VNR as an important area for the protection of amphibian and reptile biodiversity in the Atlantic Forest.(AU)

Phylogenetic position of Gorgoderina parvicava Travassos, 1922 (Digenea: Gorgoderidae), a parasite of Leptodactylus labyrinthicus (Spix, 1824) (Anura: Leptodactylidae) in Brazil / Posição filogenética de Gorgoderina parvicava Travassos, 1922 (Digenea: Gorgoderidae), parasita de Leptodactylus labyrinthicus (Spix, 1824) (Anura: Leptodactylidae) no Brasil

During a parasite survey in Brazilian amphibians from São Paulo state, Brazil, Gorgoderina parvicava Travassos, 1922 was found in the urinary bladder (11 adult worms) and (five juvenile worms) in the kidneys of the pepperfrog Leptodactylus labyrinthicus (Spix, 1824). Parasites were examined by microscopy and 28S rDNA and COI gene were sequenced and analyzed for the molecular study. The phylogenetic reconstructions resulted in identical topologies with highly supported values in the nodes in most clades using Maximum likelihood and Bayesian inference methods and positioned G. parvicava in the subclade formed by species of subfamily Gorgoderinae parasitizing the urinary bladder of amphibians. Molecular phylogenetic data showed that this species is related to other species of Gorgoderina. In addition, new molecular data and the analyses of genetic distances provide extra comparative data, which can be applied in further investigations on the taxonomic status and the diversity among Gorgoderina spp. and host-parasite relationships.

Durante o levantamento de parasitas de anfíbios brasileiros do estado de São Paulo, Brasil, Gorgoderina parvicava Travassos, 1922 foi encontrado na bexiga urinária (11 vermes adultos) e nos rins (cinco vermes juvenis) da rãpimenta Leptodactylus labyrinthicus (Spix, 1824). Os parasitas foram examinados por microscopia e os genes 28S rDNA e COI foram sequenciados e analisados para o estudo molecular. As reconstruções filogenéticas resultaram em topologias idênticas com valores suportados nos nós na maioria dos clados, usando métodos de máxima verossimilhança e inferência bayesiana e posicionaram G. parvicava no subclado formado por espécies da subfamília Gorgoderinae parasitando a bexiga urinária de anfíbios. Dados filogenéticos moleculares mostraram que esta espécie está relacionada a outras espécies de Gorgoderina. Além disso, novos dados moleculares e as análises de distâncias genéticas fornecem dados comparativos extras, que podem ser aplicados em futuras investigações sobre o status taxonômico e a diversidade entre Gorgoderina spp. e relações parasita-hospedeiro.

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.(AU)

Self-made home: how and where does the anuran Rhinella dorbignyi build its retreat sites / Com as próprias patas: Como e onde o Sapinho-de-Jardim Rhinella dorbignyi constrói seus abrigos

In this study, we observed that burrows of Rhinella dorbignyi (Duméril & Bibron, 1841) are distributed in a non-random manner in the habitat, suggesting a microhabitat selection for digging. This conclusion was based on a characterization of 36 burrows and surrounding micro-habitat. We established a 1 m x 1 m quadrat with the burrow in its central point (n=36) to measure the percentage (density) and the average heights of grasses, herbs, and shrubs. All measurements were repeated in two unused quadrats (without burrows) to evaluate the available microhabitat (n=72). The burrows are built in specific areas of the habitat with a higher percentage of grass, taller herbs, lower density of shrubs and low shaded sites than the founded at control sites. Based on three-dimensional models of the interior of the burrow (n=15), we observed that all of them were constructed with an elliptical opening that opens into a narrow channel perpendicular to the ground. Channels had a mean maximum diameter of 38 mm and a mean minimum diameter of 18 mm. The mean length of the burrows is 182 mm, and the mean volume is 95 mL.

Neste estudo, observamos que tocas de Rhinella dorbignyi (Duméril & Bibron, 1841) são distribuídas de uma forma não-aleatória no habitat, sugerindo que esta espécie seleciona sítios para cavá-las. Esta conclusão foi baseada em uma caracterização de 36 tocas e do micro-habitat que as cerca. Estabelecemos um quadrante de 1 m x 1 m com a toca como seu ponto central (n=36) para medir a porcentagem (densidade) e a altura média de gramíneas, plantas herbáceas e arbustos. Todas as medidas foram repetidas em dois quadrantes não utilizados (sem tocas), para avaliar o micro-habitat disponível ao anfíbio (n=72). As tocas são construídas em áreas específicas do habitat com maior porcentagem de gramíneas, ervas mais altas, menos arbustos presentes e pouca área sombreada. Baseado em modelos tridimensionais do interior das tocas (n=15), observamos que todas elas são construídas com uma abertura elíptica que se abre para um canal estreito, perpendicular ao chão. Os canais têm diâmetro máximo médio de 38 mm e diâmetro mínimo médio de 18 mm. O comprimento médio das tocas é de 182 mm, e o volume médio é de 95 mL.

Helminth's assemblage of a small frog in the Brazilian semiarid: parasite-host-environment relationships / Assembleia de helmintos em uma pequena rã no semiárido brasileiro: relação parasito-hospedeiro-ambiente

Parasite infections may contingency different aspects of the hosts' lives, and are usually expected to directly or indirectly compromise host reproduction. However, although amphibians have historically been study models in parasite ecology, the effects of parasitism on amphibians' reproduction remain unknown. Thus, we investigated how the parasite load varies as a function of climatic seasonality (rainy and dry season) and intrinsic host characteristics (size and sex), as well as the relationship between parasite load and reproductive investment in males and females of a small frog (Pseudopaludicola pocoto Magalhães, Loebmann, Kokubum, Haddad & Garda, 2014) in the Brazilian semiarid region. The parasitological parameters of the studied population were not influenced by the season of the year or by the hosts' body size, but females of P. pocoto had a higher prevalence and intensity of infection than males. The number of oocytes and the volume of the testes were not related to the parasite load, revealing that the parasitism did not negatively impact the gonadal investment in P. pocoto. Our findings suggest that short-lived species, such as the tiny Pseudopaludicola species, have a high reproductive investment independent of their parasitic interactions. In addition, this should be true mainly in those species that live in seasonally dry environments, such as P. pocoto, in which reproduction is even more constrained by the shortened and unpredictable rainy period.

Como infecções parasitárias podem contigenciar diversos aspectos da vida dos hospedeiros, de modo geral, espera-se que o parasitismo comprometa direta ou indiretamente a reprodução dos hospedeiros. Contudo, apesar dos anfíbios historicamente terem sido modelos de estudos em ecologia parasitária, os efeitos do parasitismo na reprodução de anfíbios permanecem incompreendidos. Neste trabalho investigamos como a carga parasitária varia em função da sazonalidade climática (período chuvoso e seco) e de características intrínsecas dos hospedeiros (tamanho e sexo), bem como a relação entre a intensidade de infecção e o investimento reprodutivo em machos e fêmeas de uma pequena rã (Pseudopaludicola pocoto Magalhães, Loebmann, Kokubum, Haddad & Garda, 2014) no semiárido brasileiro. Os parâmetros parasitológicos da população estudada não foram influenciados pela estação do ano nem pelo tamanho corpóreo dos hospedeiros, mas as fêmeas de P. pocoto apresentaram maior prevalência e intensidade de infecção que os machos. A carga parasitária não foi relacionada com o número de ovócitos nem com o volume dos testículos, revelando que o parasitismo não impactou negativamente o investimento gonadal em P. pocoto. Nossos achados sugerem que espécies com baixa longevidade, como as diminutas espécies de Pseudopaludicola, apresentam um alto investimento reprodutivo independente de suas interações parasitárias. Complementarmente, isto deve ocorrer sobretudo naquelas espécies que vivem em ambientes sazonalmente secos, como P. pocoto, nos quais a reprodução é ainda mais contigenciada pela brevidade e imprevisibilidade do período chuvoso.

Impact of atrazine and nitrate on liver and kidney of egyptian toad Sclerophrys regularis: bioindicator alarming on ecosystem

Atrazine and nitrate have been shown to act as potent oxidative stressors in amphibians either alone or in combination under stable laboratory conditions, causing histopathological alternations in liver and kidney structures at the sub-lethal concentrations. A control group and three treatments groups were tested; atrazine, nitrate, atrazine-nitrate treatments with doses of 300 µg L-1, 200 mg L-1 and their combination respectively. Sever distortion in liver and kidney tissues were shown related to the different treatments. The most hepatic lesions were observed depletion in glycogen content, degeneration of hepatocytes, hemorrhage, necrosis, vasodilatation, congestion in blood vessels, cloudy swelling in the hepatocytes and aggregation of melanomacrophage cells in between the hepatocytes that increased in combination treatment group. In kidney, the most lesions were represented in degeneration of renal tubules, fibrosis, hemorrhage, leucocytes infiltration, thickness in the wall of the renal capsule, atrophy of glomerulus, deformation of Bowman's epithelium. These negative impacts may be a bioindicator alarming the ecosystem disrupting caused by the uncontrolled apply of these chemicals in agriculture.(AU)

Identification key for anuran amphibians in a protected area in the northeastern Atlantic Forest

Abstract The identification of anuran amphibians is still a challenge in megadiverse assemblages. In the Neotropics, the Atlantic Forest harbors more than 600 anuran species, and many studies in this ecoregion report anuran assemblages surpassing 30 species. Taxonomic keys facilitate the identification of biological diversity, however only a few are available for anuran assemblages in the Atlantic Forest. Herein we present an identification key for 40 anuran species distributed across 20 genera and nine families, occurring in the Environmental Protection Area of Catolé and Fernão Velho, northeastern Atlantic Forest. Thirty-five morphological characteristics were used in the key, all of which can be easily observed in living and museum specimens. This pioneer study provides the first identification key for an amphibian assemblage in the northeastern Atlantic Forest and this baseline information acts as the starting point for the development of evolutionary and ecological research in this conservation unit.

Identification key for anuran amphibians in a protected area in the northeastern Atlantic Forest

Abstract The identification of anuran amphibians is still a challenge in megadiverse assemblages. In the Neotropics, the Atlantic Forest harbors more than 600 anuran species, and many studies in this ecoregion report anuran assemblages surpassing 30 species. Taxonomic keys facilitate the identification of biological diversity, however only a few are available for anuran assemblages in the Atlantic Forest. Herein we present an identification key for 40 anuran species distributed across 20 genera and nine families, occurring in the Environmental Protection Area of Catolé and Fernão Velho, northeastern Atlantic Forest. Thirty-five morphological characteristics were used in the key, all of which can be easily observed in living and museum specimens. This pioneer study provides the first identification key for an amphibian assemblage in the northeastern Atlantic Forest and this baseline information acts as the starting point for the development of evolutionary and ecological research in this conservation unit.

Celiotomia e gastrotomia para remoção cirúrgica de corpos estranhos em axolote (Ambystoma mexicanum): relato de caso / Celiotomy and gastrotomy for surgical removal of foreign bodies in axolotl (Ambystoma mexicanum): case report

A ingestão de corpos estranhos em aquários artificiais é uma ocorrência frequente observada na clínica cirúrgica de anfíbios criados como pets, o que inclui os axolotes (Ambystoma mexicanum). O presente relato descreve um caso de ingestão de sete corpos estranhos em um axolote macho, de cinco meses de idade, com histórico de abaulamento irregular e irredutível da superfície corpórea ventral, de consistência firme. Na anamnese foi informado que o animal habitava um aquário com substrato de cascalhos. Ao exame físico, o axolote apresentou bom estado geral e parâmetros vitais dentro da normalidade para a espécie. Os materiais deglutidos foram identificados em região gástrica por meio de exame radiográfico corpóreo total, e suas características condiziam com o substrato utilizado no aquário do paciente. Como protocolo anestésico, foi priorizada a imersão em Isofluorano e gás oxigênio, com o objetivo de atingir a via branquial e, ocasionalmente, transdérmica. A remoção cirúrgica foi feita através de celiotomia e gastrotomia em ambiente aquático com temperatura, pH e coleção bacteriana controlados, conforme literatura disponível e tendo em consideração a natureza, diâmetro e localização dos corpos estranhos. Após cinco e quinze dias do procedimento, acompanhou-se a cicatrização da ferida cirúrgica, sendo possível constatar bom restabelecimento da continuidade dos tecidos e bom estado geral do paciente.(AU)

The ingestion of foreign bodies in artificial aquariums is a frequent occurrence observed in the surgical clinic of amphibians raised as pets, which includes axolotls (Ambystoma mexicanum). The present report describes a case of ingestion of seven foreign bodies in a male axolotl, five months old, with a history of irregular and irreducible bulging of the ventral body surface, with firm consistency. In the anamnesis, it was reported that the animal inhabited an aquariums with gravel substrate. On physical examination, the axolotl showed good general condition and vital parameters within the normal range for the species. The swallowed materials were identified in the gastric region by means of total body radiographic examination, and their characteristics were consistent with the substrate used in the patients aquarium. As anesthetic protocol, immersion in Isofluorane and oxygen gas was prioritized, in order to reach the branchial and, occasionally, transdermal route. Surgical removal was performed through celiotomy and gastrotomy in an aquatic environment with controlled temperature, pH and bacterial collection, according to available literature and taking into account the nature, diameter and location of foreign bodies. After five and fifteen days of the procedure, the healing of the surgical wound was monitored, showing a good restoration of tissue continuity and a good general condition of the patient.(AU)

Celiotomia e gastrotomia para remoção cirúrgica de corpos estranhos em axolote (Ambystoma mexicanum): relato de caso / Celiotomy and gastrotomy for surgical removal of foreign bodies in axolotl (Ambystoma mexicanum): case report

A ingestão de corpos estranhos em aquários artificiais é uma ocorrência frequente observada na clínica cirúrgica de anfíbios criados como pets, o que inclui os axolotes (Ambystoma mexicanum). O presente relato descreve um caso de ingestão de sete corpos estranhos em um axolote macho, de cinco meses de idade, com histórico de abaulamento irregular e irredutível da superfície corpórea ventral, de consistência firme. Na anamnese foi informado que o animal habitava um aquário com substrato de cascalhos. Ao exame físico, o axolote apresentou bom estado geral e parâmetros vitais dentro da normalidade para a espécie. Os materiais deglutidos foram identificados em região gástrica por meio de exame radiográfico corpóreo total, e suas características condiziam com o substrato utilizado no aquário do paciente. Como protocolo anestésico, foi priorizada a imersão em Isofluorano e gás oxigênio, com o objetivo de atingir a via branquial e, ocasionalmente, transdérmica. A remoção cirúrgica foi feita através de celiotomia e gastrotomia em ambiente aquático com temperatura, pH e coleção bacteriana controlados, conforme literatura disponível e tendo em consideração a natureza, diâmetro e localização dos corpos estranhos. Após cinco e quinze dias do procedimento, acompanhou-se a cicatrização da ferida cirúrgica, sendo possível constatar bom restabelecimento da continuidade dos tecidos e bom estado geral do paciente.

The ingestion of foreign bodies in artificial aquariums is a frequent occurrence observed in the surgical clinic of amphibians raised as pets, which includes axolotls (Ambystoma mexicanum). The present report describes a case of ingestion of seven foreign bodies in a male axolotl, five months old, with a history of irregular and irreducible bulging of the ventral body surface, with firm consistency. In the anamnesis, it was reported that the animal inhabited an aquariums with gravel substrate. On physical examination, the axolotl showed good general condition and vital parameters within the normal range for the species. The swallowed materials were identified in the gastric region by means of total body radiographic examination, and their characteristics were consistent with the substrate used in the patient’s aquarium. As anesthetic protocol, immersion in Isofluorane and oxygen gas was prioritized, in order to reach the branchial and, occasionally, transdermal route. Surgical removal was performed through celiotomy and gastrotomy in an aquatic environment with controlled temperature, pH and bacterial collection, according to available literature and taking into account the nature, diameter and location of foreign bodies. After five and fifteen days of the procedure, the healing of the surgical wound was monitored, showing a good restoration of tissue continuity and a good general condition of the patient.

In vitro antimicrobial activity of methanolic extracts from cutaneous secretions of Amazonian amphibians against phytopathogens of agricultural interest / Atividade antimicrobiana in vitro de extratos metanólicos de secreções cutâneas de anfíbios amazônicos sobre fitopatógenos de interesse agrícola

The biochemical defense mechanisms of amphibians involve cutaneous secretions of bioactive molecules with antimicrobial activity. This study evaluated the in vitro activity of methanolic extracts from cutaneous secretions of two amphibian species of the Bufonidae family, Rhaebo guttatus and Rhinella marina, in the control of the phytopathogens Fusarium udum, Fusarium solani, Colletotrichum truncatum, Aspergillus flavus, Rhizoctonia solani, Macrophomina phaseolina, and Calonectria pseudometrosideri. The R. guttatus extract decreased the mycelial growth of F. udum, F. solani, A. flavus, and M. phaseolina at some tested concentrations. The R. marina extract decreased the mycelial growth of C. truncatum at the concentration of 0.5 mg mL-¹, and inhibited the mycelial growth of A. flavus at concentrations of 0.1 and 0.5 mg mL-¹, which was similar to the inhibition by the positive control. The R. marina extract also decreased the microsclerotia production by R. solani at concentrations of 0.2 and 0.3 mg mL-¹. In addition, the extracts inhibited conidial sporulation and germination at varying degrees. The inhibition of appressoria formation in C. truncatum by the R. guttatus and R. marina extracts was 85-99% and 63-100%, respectively. Our results demonstrated that treatment with extracts from R. guttatus and R. marina cutaneous secretions showed antifungal activity against the studied phytopathogens.(AU)

Os mecanismos de defesa bioquímica dos anfíbios envolvem secreções cutâneas de moléculas bioativas com atividade antimicrobiana. Este estudo avaliou a atividade in vitro de extratos metanólicos da secreção cutânea de duas espécies de anfíbios da família Bufonidae, Rhaebo guttatus e Rhinella marina, no controle dos patógenos Fusarium udum, Fusarium solani, Colletotrichum truncatum, Aspergillus flavus, Rhizoctonia solani, Macrophomina phaseolina e Calonectria pseudometrosideri. O extrato de R. guttatus inibiu o crescimento micelial de F. udum, F. solani, A. flavus e M. phaseolina em algumas concentrações testadas. O extrato de R. marina inibiu o crescimento micelial de C. truncatum na concentração de 0,5 mg mL-¹, e inibiu o crescimento micelial de A. flavus nas concentrações de 0,1 e 0,5 mg mL-¹, que foi semelhante à inibição pelo controle positivo. O extrato de R. marina também diminuiu a produção de microescleródios de R. solani nas concentrações de 0,2 e 0,3 mg mL-¹. Além disso, os extratos inibiram a esporulação e germinação de conídios em graus variados. A inibição da formação de apressórios em C. truncatum pelos extratos de R. guttatus e R. marina foi de 85%-99% e 63%-100%, respectivamente. Nossos resultados demonstraram que o tratamento com extratos da secreção cutânea de R. guttatus e R. marina apresentou atividade antifúngica sobre os fitopatógenos estudados.(AU)