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)

Intoxicação de cães por venenos de anuros: prevalência, danos aos animais e protocolos clínicos / Dog poisoning by anuran poisons: prevalence, animal damage and clinical protocols

Indivíduos da família Bufonidae são comumente associados a casos de intoxicação de cães após contato com o veneno secretado nas glândulas presentes no tegumento, reforçando a importância de tratamentos efetivos após esta interação. Neste trabalho apresentamos uma revisão sobre casos de intoxicação de cães por venenos de anuros, relacionando os principais sintomas nos cães e os protocolos clínicos utilizados. Realizamos uma busca por artigos que relatem casos de intoxicação de cães por anuros nos repositórios Google Scholar, Scielo e PubMed. Utilizamos palavras-chave nos idiomas português e inglês. Um total de 430 artigos foram encontrados, sendo apenas 17 de acordo com a proposta da pesquisa. Os registros encontrados foram para o Brasil, Austrália e Estados Unidos. As espécies de anuros reportadas na literatura foram exclusivamente as do gênero Rhinella. O maior número dos casos registrados no Brasil ocorreu em ambiente urbano. Os principais sintomas descritos após intoxicação de cães foram salivação em excesso, convulsão e vômito. Óbitos também foram encontrados durante a busca. Principais protocolos para o tratamento após envenenamento foram lavagem da cavidade oral do cão, e administração de atropina, diazepam e fluidoterapia. Foi observado que há influência do tamanho dos cães na severidade após intoxicação, sendo os de pequeno porte mais suscetíveis a quadros letais. Sugerimos, por conta do baixo número de registros, que possivelmente a quantidade de casos acerca desta temática seja subestimado. Neste estudo evidenciamos que os protocolos utilizados para o cuidado dos cães intoxicados não são realizados de forma padrão, alterando de acordo com o quadro clínico apresentado.

Individuals of the Bufonidae family are commonly associated with cases of poisoning in dogs after contact with the venom secreted in glands present in the tegument, reinforcing the importance of effective treatments after this interaction. In this paper, we present a review of cases of poisoning in dogs by anuran venom, relating the main symptoms in dogs and the clinical protocols used. We performed a search for articles reporting cases of anuran poisoning in dogs in the Google Scholar, Scielo, and PubMed repositories. We used keywords in Portuguese and English. A total of 430 articles were found and only 17 of which were in accordance with the research proposal. The records found were for Brazil, Australia, and the United States. The species of anurans reported in the literature were exclusively those of the genus Rhinella. The greatest number of cases registered in Brazil occurred in an urban environment. The main symptoms described after intoxication in dogs were excessive salivation, convulsion, and vomiting. Deaths were also found during the search. The main protocols for treatment after poisoning were washing the dog's oral cavity, and administration of atropine, diazepam, and fluid therapy. It was observed that the size of the dogs influences the severity after intoxication, with small dogs being more susceptible to lethal conditions. Due to the low number of records, we suggest that the number of cases on this topic is possibly underestimated. In this study, we showed that the protocols used for the care of intoxicated dogs are not performed in a standard way, changing according to the clinical picture presented.

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)

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)

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)

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)

Individual variation in the advertisement call of Aplastodiscus albosignatus (Anura: Hylidae) is correlated with body size and environmental temperature

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)

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.

El rol del sexo en la estructura de la población de nematodes en una especie de anfibio del Monte de Argentina

RESUMEN Diversos estudios comparativos han demostrado que los hospedadores machos no sólo podrían albergar más parásitos que las hembras, sino también tener un rol en la estructura de la población de nematodes. Debido a esto, debatimos la función que tiene el sexo en el parasitismo en una población de anfibios en un sector del monte. Analizamos el contenido gastrointestinal en busca de endoparásitos de 43 ejemplares de Pleurodema nebulosum (Burmeister, 1861), 38 adultos (26 machos y 12 hembras) y 5 juveniles. Nuestros resultados indicaron que únicamente machos se encontraban parasitados asociado con una curva correspondiente a un modelo parasitismo/ rango - edad tipo III. Aplectana nebulosa Piñeiro-Gómez, González & Sanabria, 2017 (Nematoda: Cosmocercidae) presenta un patrón de distribución agregado ajustado a un modelo Poisson. Pleurodema nebulosum (Anura: Leptodactylidae) es oportunista para reproducirse en periodos de lluvia, debido a esto nosotros proponemos como hipótesis que el momento de infección por nematodes probablemente ocurra durante estos cortos periodos en lagunas temporales formadas. El parasitismo en machos podría estar relacionado a factores hormonales debido al periodo reproductivo. Por lo tanto el sexo sería un factor importante en la estructuración de las poblaciones parásitas de esta especie de anfibio.

ABSTRACT The role of sex in the structure of the nematode population in an amphibian species from Monte de Argentina. Several comparative studies demonstrated that male host could have more parasites than female, in a way that host sex could play a role in the structure of the nematode parasite populations. Here we discussed the role of sex of parasitism in an amphibian population in a sector of the Monte. We analyzed the gastrointestinal content for endoparasites of 43 Pleurodema nebulosum (Burmeister, 1861) specimens, 38 adults (26 males and 12 females) and 5 juveniles. Our results indicated that only males were parasitized associated with a curve corresponding to a type III model. Aplectana nebulosa Piñeiro-Gómez, González & Sanabria, 2017 (Nematoda: Cosmocercidae) presents an aggregate distribution pattern adjusted to a Poisson model. Pleurodema nebulosum (Anura: Leptodactylidae) is opportunistic to reproduce in periods of rain, because of this we propose the hypothesis that the moment of infection by nematodes probably occurs during these short periods in temporary lagoons formed. Parasitism in males may be related to hormonal factors due to the reproductive period. Therefore, sex would be an important factor that structures parasitism in this species of amphibian.

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.

El rol del sexo en la estructura de la población de nematodes en una especie de anfibio del Monte de Argentina / The role of sex in the structure of the nematode population in an amphibian species from Monte de Argentina

Diversos estudios comparativos han demostrado que los hospedadores machos no sólo podrían albergar más parásitos que las hembras, sino también tener un rol en la estructura de la población de nematodes. Debido a esto, debatimos la función que tiene el sexo en el parasitismo en una población de anfibios en un sector del monte. Analizamos el contenido gastrointestinal en busca de endoparásitos de 43 ejemplares de Pleurodema nebulosum (Burmeister, 1861), 38 adultos (26 machos y 12 hembras) y 5 juveniles. Nuestros resultados indicaron que únicamente machos se encontraban parasitados asociado con una curva correspondiente a un modelo parasitismo/ rango - edad tipo III. Aplectana nebulosa Piñeiro-Gómez, González & Sanabria, 2017 (Nematoda: Cosmocercidae) presenta un patrón de distribución agregado ajustado a un modelo Poisson. Pleurodema nebulosum (Anura: Leptodactylidae) es oportunista para reproducirse en periodos de lluvia, debido a esto nosotros proponemos como hipótesis que el momento de infección por nematodes probablemente ocurra durante estos cortos periodos en lagunas temporales formadas. El parasitismo en machos podría estar relacionado a factores hormonales debido al periodo reproductivo. Por lo tanto el sexo sería un factor importante en la estructuración de las poblaciones parásitas de esta especie de anfibio.(AU)

The role of sex in the structure of the nematode population in an amphibian species from Monte de Argentina. Several comparative studies demonstrated that male host could have more parasites than female, in a way that host sex could play a role in the structure of the nematode parasite populations. Here we discussed the role of sex of parasitism in an amphibian population in a sector of the Monte. We analyzed the gastrointestinal content for endoparasites of 43 Pleurodema nebulosum (Burmeister, 1861) specimens, 38 adults (26 males and 12 females) and 5 juveniles. Our results indicated that only males were parasitized associated with a curve corresponding to a type III model. Aplectana nebulosa Piñeiro-Gómez, González & Sanabria, 2017 (Nematoda: Cosmocercidae) presents an aggregate distribution pattern adjusted to a Poisson model. Pleurodema nebulosum (Anura: Leptodactylidae) is opportunistic to reproduce in periods of rain, because of this we propose the hypothesis that the moment of infection by nematodes probably occurs during these short periods in temporary lagoons formed. Parasitism in males may be related to hormonal factors due to the reproductive period. Therefore, sex would be an important factor that structures parasitism in this species of amphibian.(AU)

El rol del sexo en la estructura de la población de nematodes en una especie de anfibio del Monte de Argentina / The role of sex in the structure of the nematode population in an amphibian species from Monte de Argentina

Diversos estudios comparativos han demostrado que los hospedadores machos no sólo podrían albergar más parásitos que las hembras, sino también tener un rol en la estructura de la población de nematodes. Debido a esto, debatimos la función que tiene el sexo en el parasitismo en una población de anfibios en un sector del monte. Analizamos el contenido gastrointestinal en busca de endoparásitos de 43 ejemplares de Pleurodema nebulosum (Burmeister, 1861), 38 adultos (26 machos y 12 hembras) y 5 juveniles. Nuestros resultados indicaron que únicamente machos se encontraban parasitados asociado con una curva correspondiente a un modelo parasitismo/ rango - edad tipo III. Aplectana nebulosa Piñeiro-Gómez, González & Sanabria, 2017 (Nematoda: Cosmocercidae) presenta un patrón de distribución agregado ajustado a un modelo Poisson. Pleurodema nebulosum (Anura: Leptodactylidae) es oportunista para reproducirse en periodos de lluvia, debido a esto nosotros proponemos como hipótesis que el momento de infección por nematodes probablemente ocurra durante estos cortos periodos en lagunas temporales formadas. El parasitismo en machos podría estar relacionado a factores hormonales debido al periodo reproductivo. Por lo tanto el sexo sería un factor importante en la estructuración de las poblaciones parásitas de esta especie de anfibio.

The role of sex in the structure of the nematode population in an amphibian species from Monte de Argentina. Several comparative studies demonstrated that male host could have more parasites than female, in a way that host sex could play a role in the structure of the nematode parasite populations. Here we discussed the role of sex of parasitism in an amphibian population in a sector of the Monte. We analyzed the gastrointestinal content for endoparasites of 43 Pleurodema nebulosum (Burmeister, 1861) specimens, 38 adults (26 males and 12 females) and 5 juveniles. Our results indicated that only males were parasitized associated with a curve corresponding to a type III model. Aplectana nebulosa Piñeiro-Gómez, González & Sanabria, 2017 (Nematoda: Cosmocercidae) presents an aggregate distribution pattern adjusted to a Poisson model. Pleurodema nebulosum (Anura: Leptodactylidae) is opportunistic to reproduce in periods of rain, because of this we propose the hypothesis that the moment of infection by nematodes probably occurs during these short periods in temporary lagoons formed. Parasitism in males may be related to hormonal factors due to the reproductive period. Therefore, sex would be an important factor that structures parasitism in this species of amphibian.

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.

Quantity - but not diversity - of secreted peptides and proteins increases with age in the tree frog Pithecopus nordestinus

Amphibians inhabit the terrestrial environment, a conquest achieved after several evolutionary steps, which were still insufficient to make them completely independent of the aquatic environment. These processes gave rise to many morphological and physiological changes, making their skin (and cutaneous secretion) rich in bioactive molecules. Among the tree frogs, the secretion is composed mainly of peptides; but alkaloids, proteins and steroids can also be found depending on the species. The most known class of biologically active molecules is the antimicrobial peptides (AMPs) that act against bacteria, fungi and protozoans. Although these molecules are well-studied among the hylids, AMPs ontogeny remains unknown. Therefore, we performed peptidomic and proteomic analyses of Pithecopus nordestinus (formerly Phyllomedusa nordestina) in order to evaluate the peptide content in post-metamorphosed juveniles and adult individuals. Methods: Cutaneous secretion of both life stages of individuals was obtained and analyzed by LC-MS/MS after reduction and alkylation of disulfide bonds or reduction, alkylation and hydrolysis by trypsin. Results: Differences in the TIC profile of juveniles and adults in both treatments were observed. Moreover, the proteomic data revealed known proteins and peptides, with slight differences in the composition, according to the life stage and the treatment. AMPs were identified, and bradykinin-potentiating peptides were observed in trypsin-treated samples, which suggests a protein source of such peptide (cryptide). Conclusion: In general, skin secretion contents were similar between juveniles and adults, varying in quantity, indicating that the different stages of life are reflected in the number of molecules and not on their diversity.(AU)

A summary of reptile and anuran amphibian species from Brazilian sandy coastal plains: 31 years of sampling efforts of the Laboratório de Ecologia de Vertebrados, Universidade do Estado do Rio de Janeiro / Um resumo das espécies de répteis e anfíbios anuros das planícies costeiras arenosas brasileiras: 31 anos de esforços de amostragem do Laboratório de Ecologia de Vertebrados da Universidade do Estado do Rio de Janeiro

Although currently there is already a set of studies regarding ecological aspects of some particular reptile and amphibian species living in Brazilian sandy coastal plains (including the so-called restinga and campo nativo habitats), there is comparatively few information on the species composition usually associated to these environments. During 31 years (1988-2019) of herpetological studies carried out in sandy coastal plains environments by our research team of the Laboratory of Vertebrate Ecology (Department of Ecology, Universidade do Estado do Rio de Janeiro, in Rio de Janeiro Brazil) we have surveyed reptile and amphibian communities and performed different studies with similar methods in 70 sites from 10 different states along the Brazilian coast. Our surveys resulted in records of 87 species of reptile (five turtles, two crocodylians, six amphisbaenians, 36 lizards and 39 snakes) from 24 families, and 77 species of anuran amphibians from nine families. We have studied multiple natural history topics for anurans and reptiles which resulted in the publication of some specific ecological studies, especially regarding some species, encompassing population and community ecology, foraging and feeding habits, species activity, thermoregulation, reproduction, use of microhabitats, and parasitism by ecto and endoparasites. Our results along these three decades have also contributed for the description of four new lizard species (Ameivula nativo, Glaucomastix littoralis, G. abaetensis and G. itabaianensis). Our studies constitute an important contribution to the knowledge of the ecology of anuran amphibians and reptiles in these ecosystems, as well as to the conservation of sandy coastal plains environment. The checklist presented in this study, based on our records of sandy coastal plains herpetofauna, provides for many localities along the Brazilian coast, the needed knowledge on species occurrence, including the presence of endemic and/or endangered species, which can be of value for many conservation actions.(AU)

Embora atualmente exista um conjunto de estudos sobre aspectos ecológicos de algumas espécies de répteis e de anfíbios que ocorrem nas planícies costeiras arenosas brasileiras (incluindo os chamados habitats de restinga e de campo nativo), há relativamente poucas informações sobre a composição de espécies geralmente associada a esses ambientes. Durante 31 anos (1988-2019) de estudos herpetológicos realizados em restingas por nossa equipe de pesquisa do Laboratório de Ecologia de Vertebrados (Departamento de Ecologia, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brasil) nós estudamos comunidades de répteis e de anfíbios e realizamos diferentes estudos com métodos semelhantes em 70 localidades de dez diferentes Estados ao longo da costa brasileira. Nossas pesquisas resultaram em registros de 87 espécies de répteis (cinco tartarugas, dois crocodilianos, seis anfisbênios, 36 lagartos e 39 serpentes) de 24 famílias, e 77 espécies de anfíbios anuros de nove famílias. Estudamos vários tópicos de história natural sobre anuros e répteis, que resultaram na publicação de alguns estudos ecológicos específicos, especialmente em relação a algumas espécies, abrangendo ecologia populacional e de comunidades, forrageamento e dieta, horário de atividade de espécies, termorregulação, reprodução, uso do microhabitat e parasitismo por ecto e endoparasitas. Nossos resultados ao longo dessas três décadas também contribuíram para a descrição de quatro novas espécies de lagartos (Ameivula nativo, Glaucomastix littoralis, G. abaetensis e G. itabaianensis). Nossos estudos constituem uma importante contribuição para o conhecimento da ecologia de répteis e de anfíbios anuros nesses ecossistemas, bem como para a conservação dos ecossistemas de restinga. A lista de espécies apresentada neste estudo, com base em nossos registros de herpetofauna das planícies costeiras arenosas, fornece para muitas localidades ao longo da costa brasileira o conhecimento necessário sobre a ocorrência de espécies, incluindo a presença de espécies endêmicas e/ ou ameaçadas de extinção, que podem ser úteis para muitas ações de conservação.(AU)

Quantity - but not diversity - of secreted peptides and proteins increases with age in the tree frog Pithecopus nordestinus

Amphibians inhabit the terrestrial environment, a conquest achieved after several evolutionary steps, which were still insufficient to make them completely independent of the aquatic environment. These processes gave rise to many morphological and physiological changes, making their skin (and cutaneous secretion) rich in bioactive molecules. Among the tree frogs, the secretion is composed mainly of peptides; but alkaloids, proteins and steroids can also be found depending on the species. The most known class of biologically active molecules is the antimicrobial peptides (AMPs) that act against bacteria, fungi and protozoans. Although these molecules are well-studied among the hylids, AMPs ontogeny remains unknown. Therefore, we performed peptidomic and proteomic analyses of Pithecopus nordestinus (formerly Phyllomedusa nordestina) in order to evaluate the peptide content in post-metamorphosed juveniles and adult individuals. Methods: Cutaneous secretion of both life stages of individuals was obtained and analyzed by LC-MS/MS after reduction and alkylation of disulfide bonds or reduction, alkylation and hydrolysis by trypsin. Results: Differences in the TIC profile of juveniles and adults in both treatments were observed. Moreover, the proteomic data revealed known proteins and peptides, with slight differences in the composition, according to the life stage and the treatment. AMPs were identified, and bradykinin-potentiating peptides were observed in trypsin-treated samples, which suggests a protein source of such peptide (cryptide). Conclusion: In general, skin secretion contents were similar between juveniles and adults, varying in quantity, indicating that the different stages of life are reflected in the number of molecules and not on their diversity.(AU)

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)

Selection and use of calling site by Boana leptolineata and Phyllomedusa distincta during the reproductive season / Seleção e uso de sítios de vocalização por Boana leptolineata e Phyllomedusa distincta durante o período reprodutivo

Climatic conditions and microhabitat characteristics, such as the substrate and type of vegetation, influence the choice of male anurans for calling sites that optimize their reproductive success. We evaluated the structure and selection of vocalization microhabitat of 17 individuals of Phyllomedusa distincta Lutz, 1950 and 25 of Boana leptolineata (Braun & Braun, 1977), in a subtropical forest in southern Brazil. We measure the height of the perch, the distance between the perch and the edge of the body of water and the structure of the microhabitat of quadrants used by individuals. The same attributes were measured in the quadrants available (but not used) by the individuals. We classified the microhabitats by visual estimate, assigning percentages of coverage for each quadrant by herbaceous, shrub, tree and wetland vegetation. We observed that both species selected their microhabitat, since the characteristics of the quadrants occupied by the individuals were different from those available. Phyllomedusa distincta was more associated with heterogeneous microhabitats, while B. leptolineata occurred in environments with greater coverage of tree strata. Additionally, we observed that both species used shrubs more frequently as a perch site. The selection of these microhabitat characteristics must be associated with strategies to optimize the use of the reproductive habitat, based on the morphological and behavioral characteristics of the species. Finally, it is possible to infer that the differences observed in the microhabitat structure selected by the species can facilitate the coexistence of both in the context of the heterogeneity of the environment.(AU)

Seleção e uso de sítios de vocalização por Boana leptolineata e Phyllomedusa distincta durante o período reprodutivoAs condições climáticas e as características de micro-habitat, como o substrato e o tipo de vegetação, influenciam a escolha de anuros machos por sítios de vocalização que otimizem seu sucesso reprodutivo. Avaliamos a estrutura e a seleção de micro-habitat de vocalização de 17 indivíduos de Phyllomedusa distincta Lutz, 1950 e 25 de Boana leptolineata (Braun & Braun, 1977), em uma floresta subtropical no sul do Brasil. Medimos a altura do poleiro, a distância do poleiro até a margem do corpo dágua e a estrutura do micro-habitat de quadrantes utilizados pelos indivíduos. Os mesmos atributos foram mensurados nos quadrantes disponíveis (mas não utilizados) pelos indivíduos. Classificamos os micro-habitats por estimativa visual, atribuindo-se porcentagens de cobertura de cada quadrante por vegetação herbácea, arbustiva, arbórea e de área alagada. Observamos que ambas as espécies selecionaram seu micro-habitat, uma vez que as características dos quadrantes ocupados pelos indivíduos foram distintas daquelas disponíveis. Phyllomedusa distincta esteve mais associada a micro-habitats heterogêneos, enquanto B. leptolineata ocorreu em ambientes com maior cobertura de estrato arbóreo. Adicionalmente, observamos que ambas as espécies utilizaram mais frequentemente arbustos como local do poleiro. A seleção destas características de micro-habitat deve estar associada a estratégias para otimização do uso do habitat reprodutivo, baseadas em características morfológicas e comportamentais das espécies. Por fim, é possível inferir que as diferenças observadas na estrutura de micro-habitat selecionadas pelas espécies podem facilitar a coexistência de ambas no contexto da heterogeneidade do ambiente.(AU)