Building pondscapes for amphibian metapopulations.

The success of ponds constructed to restore ecological infrastructure for pond-breeding amphibians and benefit aquatic biodiversity depends on where and how they are built. We studied effects of pond and landscape characteristics, including connectivity, on metapopulation dynamics of 12 amphibian species in Switzerland. To understand the determinants of long-term occupancy (here summarized as incidence), environmental effects on both colonization and persistence should be considered. We fitted dynamic occupancy models to 20 years of monitoring data on a pond construction program to quantify effects of pond and landscape characteristics and different connectivity metrics on colonization and persistence probabilities in constructed ponds. Connectivity to existing populations explained dynamics better than structural connectivity metrics, and simple metrics (distance to the nearest neighbor population, population density) were useful surrogates for dispersal kernel-weighted metrics commonly used in metapopulation theory. Population connectivity mediated the persistence of conservation target species in new ponds, suggesting source-sink dynamics in newly established populations. Population density captured this effect well and could be used by practitioners for site selection. Ponds created where there were 2-4 occupied ponds within a radius of ∼0.5 km had >3.5 times higher incidence of target species (median) than isolated ponds. Species had individual preferences regarding pond characteristics, but breeding sites with larger (≥100 m2) total water surface area, that temporarily dried, and that were in surroundings with maximally 50% forest benefitted multiple target species. Pond diversity will foster amphibian diversity at the landscape scale.

Construcción de estanques para meta poblaciones de anfibios Resumen El éxito de los estanques construidos para restaurar la infraestructura ecológica para los anfibios que allí se reproducen y para beneficiar la biodiversidad acuática depende de en dónde y cómo se construyen. Estudiamos los efectos de las características de los estanques y el paisaje, incluida la conectividad, sobre la dinámica de las meta poblaciones de 12 especies de anfibios en Suiza. Se deben considerar los efectos ambientales sobre la colonización y la persistencia para entender las determinantes de la ocupación a largo plazo (resumida aquí como incidencia). Ajustamos los modelos dinámicos de ocupación a datos de 20 años de monitoreo de un programa de construcción de estanques para cuantificar los efectos de las características del estanque y el paisaje y las diferentes medidas de conectividad para las probabilidades de colonización y persistencia en los estanques construidos. La conectividad con las poblaciones existentes explicó mejor la dinámica que las medidas de conectividad estructural, mientras que las medidas simples (distancia a la población vecina más cercana, densidad poblacional) fueron sustitutos útiles para las medidas de dispersión ponderadas al núcleo que se usan con frecuencia en la teoría de meta poblaciones. La conectividad poblacional medió la persistencia de las especies a conservar en los estanques nuevos, lo que sugiere que hay dinámicas fuente­sumidero en las poblaciones recién establecidas. La densidad poblacional capturó muy bien este efecto y podría usarse para que los practicantes seleccionen sitios. Los estanques construidos en un radio de ≈0.5 km de dos a cuatro estanques ocupados tuvieron >3.5 más incidencia de las especies a conservar (mediana) que los estanques aislados. Las especies tuvieron preferencias individuales con respecto a las características de los estanques, aunque los sitios de reproducción con una mayor superficie total de agua (≥100 m2), que se secaban temporalmente y que estaban rodeados con un máximo de 50% de bosque beneficiaron a muchas especies a conservar. Por esto, la diversidad de estanques promoverá la diversidad de anfibios a escala de paisaje.

Effects of climate and human activity on the current distribution of amphibians in China.

In China, as elsewhere, amphibians are highly endangered. Anthropogenic environmental change has affected the distribution and population dynamics of species, and species distributions at a broad scale are strongly driven by climate and species' ability to disperse. Yet, current knowledge remains limited on how widespread human activity affects the distribution patterns of amphibians in China and whether this effect extends beyond climate. We compiled a relatively comprehensive database on the distribution of 196 amphibian species in China from the literature, public databases, and field data. We obtained 25,826 records on almost 50% of known species in China. To test how environmental factors and human activities influence the current distribution of amphibians (1960-1990), we used range filling, which is species realized ranges relative to their potential climate distribution. We used all species occurrence records to represent realized range and niche models to predict potential distribution range. To reduce uncertainty, we used 3 regression methods (beta regression, generalized boosted regression models, and random forest) to test the associations of species range filling with human activity, climate, topography, and range size. The results of the 3 approaches were consistent. At the species level, mean annual precipitation (climate) had the most effect on spatial distribution pattern of amphibians in China, followed by range size. Human activity ranked last. At the spatial level, mean annual precipitation remained the most important factor. Regions in southeastern of China that are currently moist supported the highest amphibian diversity, but were predicted to experience a decline in precipitation under climate change scenarios. Consequently, the distributions of amphibians will likely shift to the northwest in the future, which could affect future conservation efforts.

En China, como en todos lados, los anfibios están gravemente en peligro. El cambio ambiental antropogénico ha afectado la distribución y dinámica poblacional de especies, y la distribución de especies a gran escala están muy influidas por el clima y la habilidad de dispersión de las especies. Sin embargo, el conocimiento actual sigue siendo limitado sobre cómo la actividad humana generalizada afecta a los patrones de distribución de anfibios en China y si este efecto se extiende más allá del clima. A partir de literatura, bases de datos públicas y datos de campo, integramos una base datos relativamente completa sobre la distribución de 196 especies de anfibios en China. Obtuvimos 25,826 registros de casi 50% de las especies conocidas en China. Para probar cómo los factores ambientales y las actividades humanas influyen en la distribución actual de anfibios (1960-1990), utilizamos la ocupación de rango, que contrasta los rangos de distribución observada de las especies en relación con su distribución climática potencial. Utilizamos los registros de ocurrencia de todas las especies para representar el rango observado y modelos de nicho para predecir el rango de distribución potencial. Para reducir la incertidumbre, utilizamos 3 métodos de regresión (regresión beta, modelos de regresión acelerada generalizada y bosque aleatorio) para probar las asociaciones de la ocupación de rango de especies con la actividad humana, clima, topografía y extensión de rango. Los resultados de los tres métodos fueron consistentes. A nivel de especie, la precipitación media anual (clima) tuvo el mayor efecto sobre el patrón de distribución de anfibios en China, seguida por la extensión del rango. La actividad humana ocupó el último lugar. A nivel espacial, la precipitación media anual siguió como el factor más importante. Las regiones en el sureste de China que aun son húmedas sostuvieron la mayor diversidad de anfibios, pero se pronosticó que la precipitación declinará bajo escenarios de cambio climático. Consecuentemente, la distribución de anfibios muy probablemente cambiará hacia el noreste, lo cual podría afectar esfuerzos futuros de conservación.

Action of heparin and acetylcholine modulators on the neurotoxicity of the toad Rhinella schneideri (Anura: Bufonidae) in Brazil / Acción de la heparina y moduladores de acetilcolina en la neurotoxicidad del sapo Rhinella schneideri (Anura: Bufonidae) en Brasil

Abstract Introduction: Rhinella schneideri is a toad widely distributed in South America and its poison is characterized by inducing cardiotoxicity and neurotoxicity. Objective: In this work, we investigated pharmacological strategies to attenuate the peripheral neurotoxicity induced by R. schneideri poison in avian neuromuscular preparation. Methods: The experiments were carried out using isolated chick biventer cervicis preparation subjected to field stimulation for muscle twitches recordings or exposed to acetylcholine and potassium chloride for contracture responses. Results: Poison (10 μg/ml) produced complete neuromuscular blockade in chick biventer cervicis preparation within approximately 70 min incubation (times for 50 and 90 % blockade: 15 ± 3 min and 40 ± 2 min, respectively; P < 0.05, N= 5); contracture responses to exogenous acetylcholine and KCl were unaffected by poison indicating no specificity with postsynaptic receptors or myotoxicity, respectively. Poison (10 μg/ml)-induced neuromuscular blockade was not prevented by heparin (5 and 150 IU/ml) under pre- or post-treatment conditions. Incubation at low temperature (23-25 °C) abolished the neuromuscular blockade; after raising the temperature to 37 °C, the complete neuromuscular blockade was slightly slower than that seen in preparations directly incubated at 37 °C (times for 50 and 90 % blockade: 23 ± 2 min and 60 ± 2.5 min, respectively; P < 0.05, N= 4). Neostigmine (3.3 μM) did not reverse the neuromuscular blockade in BC preparation whereas 3,4-diaminopyridine (91.6 μM) produced a partial and sustained reversal of the twitch responses (29 ± 7.8 % of maximal reversal reached in approximately 40 min incubation; P < 0.05, N= 4). Conclusions: R. schneideri poison induces potent peripheral neurotoxicity in vitro which can be partially reversible by 3,4-diaminopyridine.

Resumen Introducción: Rhinella schneideri está ampliamente distribuida en Suramérica y su veneno es caracterizado por inducir cardiotoxicidad y neurotoxicidad. Objetivo: En este trabajo, investigamos estrategias farmacológicas para atenuar la neurotoxicidad periférica inducida por el veneno de R. schneideri en preparaciones neuromusculares de aves. Métodos: Los experimentos fueron realizados usando preparaciones de biventer cervicis de pollos sometidas a estimulación de campo para el registro de las contracciones musculares o expuestas a la acetilcolina y al cloruro de potasio para la respuesta contractural. Resultados: El veneno (10 µg/ml) provocó un bloqueo neuromuscular completo en las preparaciones después de aproximadamente 70 min de incubación (tiempos para 50 y 90 % de bloqueo: 15 ± 3 min y 40 ± 2 min, respectivamente; P < 0.05, N = 5); las contracturas en respuesta a la acetilcolina y el KCl exógenos no fueron afectadas por el veneno, indicando que no hay una interacción especifica con receptores postsinápticos o miotoxicidad respectivamente. El bloqueo neuromuscular causado por el veneno (10 µg/ml) no fue prevenido por la heparina (5 y 150 UI/ml) bajo condiciones pre y post-tratamiento. La incubación a bajas temperaturas (23-25 ºC) abolió el bloqueo neuromuscular; después de aumentar la temperatura a 37 ºC, el bloqueo neuromuscular total fue levemente más lento que el visto en preparaciones directamente incubadas a 37 ºC (tiempos para 50 y 90 % de bloqueo: 23 ± 2 min y 60 ± 2.5 min, respectivamente; P < 0.05, N= 4). Neostigmina (3.3 µM) no revirtió el bloqueo neuromuscular, mientras que 3.4-diaminopiridina (91.6 µM) produjo una reversión parcial y sostenida de las respuestas neuromusculares (29 ± 7.8 % de la reversión máxima alcanzada en aproximadamente 40 min de incubación; P < 0.05, N = 4). Conclusiones: El veneno de R. schneideri indujo neurotoxicidad periférica potente in vitro, el cual puede ser revertido por 3.4-diaminopiridina.

Immunohistochemical localization of vascular factors in tooth germ of amphibian (Cynops pyrrhogaster) / Localización inmunohistoquímica de factores vasculares en germen dentario de anfibios (Cynops pyrrhogaster)

SUMMARY: Vascular endothelial growth factor (VEGF) and its receptor, VEGFR-2, are known to regulate blood vessel endothelium growth. They play important role in human and rodents teeth development. In newt jaws, there are sequential developmental teeth germs following behind the mature teeth. We examined the immunohistochemical localization of VEGF and its receptor and showed the specific expression pattern of VEGF and VEGF receptor in Cynops pyrrhogaster sequential tooth development. The intensity of immunoreactivity for VEGF in the inner enamel epithelium was weaker than that in the outer enamel epithelium in the dentine matrix formation and mineralization stages. Finally, at the enameloid maturation and enamel-like matrix formation stage, immunoreactivity for VEGF in inner enamel epithelium was stronger than in the outer enamel epithelium. The intensity of immunoreactivity for VEGFR-2 was positive for the outer enamel epithelium throughout tooth development. The crown sides of the odontoblasts were stained especially strongly for VEGF and VEGFR-2 during the dentine matrix formation and mineralization stage of the enameloid maturation and enamel- like matrix formation stage. We postulate that the expression of VEGF in the inner enamel epithelium and odontoblast widely effects tooth development in newts, as well as in human and rodents.

RESUMEN: Se sabe que el factor de crecimiento endotelial vascular (VEGF) y su receptor, VEGFR-2, regulan el crecimiento del endotelio de los vasos sanguíneos. Desempeñan un papel importante en el desarrollo de los dientes humanos y de los roedores. En las mandíbulas de tritón, hay gérmenes dentales de desarrollo secuenciales que siguen a los dientes maduros. Examinamos la localización inmunohistoquímica de VEGF y su receptor y mostramos el patrón de expresión específico de VEGF y receptor de VEGF en el desarrollo secuencial de dientes de Cynops pyrrhogaster. La intensidad de la inmunorreactividad para VEGF en el epitelio interno del esmalte era más débil que en el epitelio externo del esmalte en las etapas de formación y mineralización de la matriz de dentina. Finalmente, en la etapa de maduración del esmalte y de formación de la matriz similar al esmalte, la inmunorreactividad para VEGF en el epitelio interno del esmalte fue más fuerte que en el epitelio externo del esmalte. La intensidad de la inmunorreactividad para VEGFR- 2 fue positiva para el epitelio externo del esmalte durante el desarrollo del diente. Los márgenes de la corona de los odontoblastos se tiñeron especialmente para VEGF y VEGFR-2 durante la etapa de formación de la matriz de dentina y mineralización de la etapa de maduración del esmalte y la etapa de formación de la matriz similar al esmalte. Postulamos que la expresión de VEGF en el epitelio interno del esmalte y odontoblastos afecta ampliamente el desarrollo de los dientes en tritones, así como en humanos y roedores.

Automated detection of frog calls and choruses by pulse repetition rate.

Anurans (frogs and toads) are among the most globally threatened taxonomic groups. Successful conservation of anurans will rely on improved data on the status and changes in local populations, particularly for rare and threatened species. Automated sensors, such as acoustic recorders, have the potential to provide such data by massively increasing the spatial and temporal scale of population sampling efforts. Analyzing such data sets will require robust and efficient tools that can automatically identify the presence of a species in audio recordings. Like bats and birds, many anuran species produce distinct vocalizations that can be captured by autonomous acoustic recorders and represent excellent candidates for automated recognition. However, in contrast to birds and bats, effective automated acoustic recognition tools for anurans are not yet widely available. An effective automated call-recognition method for anurans must be robust to the challenges of real-world field data and should not require extensive labeled data sets. We devised a vocalization identification tool that classifies anuran vocalizations in audio recordings based on their periodic structure: the repeat interval-based bioacoustic identification tool (RIBBIT). We applied RIBBIT to field recordings to study the boreal chorus frog (Pseudacris maculata) of temperate North American grasslands and the critically endangered variable harlequin frog (Atelopus varius) of tropical Central American rainforests. The tool accurately identified boreal chorus frogs, even when they vocalized in heavily overlapping choruses and identified variable harlequin frog vocalizations at a field site where it had been very rarely encountered in visual surveys. Using a few simple parameters, RIBBIT can detect any vocalization with a periodic structure, including those of many anurans, insects, birds, and mammals. We provide open-source implementations of RIBBIT in Python and R to support its use for other taxa and communities.

Los anuros (ranas y sapos) se encuentran dentro de los grupos taxonómicos más amenazados a nivel mundial. La conservación exitosa de los anuros dependerá de información mejorada sobre el estado y los cambios en las poblaciones locales, particularmente para las especies raras y amenazadas. Los sensores automatizados, como las grabadoras acústicas, tienen el potencial para proporcionar dicha información al incrementar masivamente la escala espacial y temporal de los esfuerzos de muestreo poblacional. El análisis de dicha información requerirá herramientas robustas y eficientes que puedan identificar automáticamente la presencia de una especie en las grabaciones de audio. Como las aves y los murciélagos, muchas especies de anuros producen vocalizaciones distintivas que pueden ser capturadas por las grabadoras acústicas autónomas y también son excelentes candidatas para el reconocimiento automatizado. Sin embargo, a diferencia de las aves y los murciélagos, todavía no se cuenta con una disponibilidad extensa de herramientas para el reconocimiento acústico automatizado de los anuros. Un método efectivo para el reconocimiento automatizado del canto de los anuros debe ser firme ante los retos de los datos reales de campo y no debería requerir conjuntos extensos de datos etiquetados. Diseñamos una herramienta de identificación de las vocalizaciones: la herramienta de identificación bioacústica basada en el intervalo de repetición (RIBBIT), el cual clasifica las vocalizaciones de los anuros en las grabaciones de audio con base en su estructura periódica. Aplicamos la RIBBIT a las grabaciones de campo para estudiar a dos especies: la rana coral boreal (Pseudacris maculata) de los pastizales templados de América del Norte y la rana arlequín variable (Atelopus varius), críticamente en peligro de extinción, de las selvas tropicales de América Central. Mostramos que RIBBIT puede identificar correctamente a las ranas corales boreales, incluso cuando vocalizan en coros con mucha superposición, y puede identificar las vocalizaciones de la rana arlequín variable en un sitio de campo en donde rara vez se le ha visto durante censos visuales. Mediante relativamente unos cuantos parámetros simples, RIBBIT puede detectar cualquier vocalización con una estructura periódica, incluyendo aquellas de muchos anuros, insectos, aves y mamíferos. Proporcionamos implementaciones de fuente abierta de RIBBIT en Python y en R para fomentar su uso para otros taxones y comunidades.

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.

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.

Manipulating water for amphibian conservation.

Amphibian populations globally are in decline. One great threat is the abstraction of water resources that alter surface-water hydrology. Conservation actions aimed at restoring or manipulating surface water are employed as a management tool, but empirical evidence on the effectiveness of these approaches is scarce. In this systematic review, we summarized the global experience of manipulating water for amphibian conservation. We explored examples of manipulating water to conserve amphibian species and communities. Approaches varied in their frequency of implementation and in their success. Extending hydroperiod to match larval requirements showed encouraging results, as did off-season drying to control predators. Spraying water into the environment showed several potential applications, but successes were limited. Despite some promising interventions, we identified few (n = 17) empirically supported examples of successful water manipulation to benefit amphibians. It is unclear whether this stems from publication bias or if it is an artifact of language selection. However, manipulating water shows some potential in amphibian conservation, particularly at sites with a proximal water source and in regions where aridity is increasing due to climate change. Regardless of the scale of the intervention or its perceived probability of success, high-quality reporting of empirical results will further understanding of how water manipulations can benefit threatened amphibian populations.

Manipulación del Agua para la Conservación de Anfibios Resumen Las poblaciones mundiales de anfibios están en declinación. Una gran amenaza es la extracción de los recursos hídricos que alteran la hidrología superficial. Las acciones de conservación enfocadas en la restauración o manipulación del agua superficial se emplean como herramientas de manejo, pero la evidencia empírica de la efectividad de estas estrategias es escasa. En esta revisión sistemática resumimos la experiencia mundial de la manipulación del agua para la conservación de anfibios. Exploramos ejemplos de la manipulación del agua para conservar especies y comunidades de anfibios. Las estrategias variaron en la frecuencia de implementación y en el éxito que tuvieron. La extensión del periodo hídrico para que cumpla con los requerimientos de las larvas mostró resultados alentadores, así como lo hizo la sequía atemporal para controlar a los depredadores. La aspersión de agua en el ambiente mostró varias aplicaciones potenciales, pero el éxito fue limitado. A pesar de algunas intervenciones prometedoras, identificamos pocos (n = 17) ejemplos con respaldo empírico de la manipulación exitosa del agua para el beneficio de los anfibios. Todavía no está claro si esto proviene de un sesgo en las publicaciones o si es un artificio de la selección del lenguaje. Sin embargo, la manipulación del agua muestra cierto potencial en la conservación de los anfibios, particularmente en sitios próximos a una fuente de agua y en regiones en donde la aridez está incrementando debido al cambio climático. Sin importar la escala de la intervención o la probabilidad de éxito percibida, la comunicación de alta calidad de los resultados empíricos hará crecer el entendimiento de cómo la manipulación del agua puede beneficiar a las poblaciones amenazadas de anfibios.

Antibacterial activity of the skin secretion of Phyllomedusa azurea (Anura: Hylidae) from the Central Brazil Cerrado / Actividad antibacterial de secreciones de la piel de Phyllomedusa azurea (Anura: Hylidae) en Cerrado, Brasil centro

Abstract Phyllomedusa azurea is a frog species well distributed geographically in South America, including Brazilian biomes as Pantanal and Cerrado. Compared with other anurans from the Phyllomedusinae family, there are few reports on the bioactive potential of skin-derived molecules from this species. In this perspective, the aim of the present study was to evaluate the in vitro antibacterial activity of skin secretion of P. azurea by detection of Minimum Inhibitory Concentration (MIC) of the growth of bacterial indicator strains and to determine if occurs a changing in the bacterial cell envelope permeability. The MIC determination was carried out by the microdilution plate method. The absorbance was measured and analyzed statistically using the t-test to compare two groups (0.05 % of significance). The impact of the crude extract on cell envelope permeability of Staphylococcus aureus ATCC 25923 was conducted by the crystal violet assay, and the absorbance was measured spectrophotometry followed by the calculation of the crystal violet uptake percentage. The specific MIC for S. aureus ATCC 25923 and Escherichia coli ATCC 25922 was 31.25 µg/mL, while for Bacillus subtilis ATCC 6633 was 125 µg/mL and Pseudomonas aeruginosa ATCC 27853 was 250 µg/mL. The treatment with crescent concentrations of frog skin secretion increased the crystal violet uptake by S. aureus ATCC 25923 cells, suggesting an action on the cell plasma membrane. The results demonstrated that the skin secretion of P. azurea presents antibacterial activity and merit further investigations to characterize the bioactive molecules.(AU)

Resumen P. azurea es una especie de rana bien distribuida geográficamente en América del Sur, que incluye biomas brasileños como Pantanal y Cerrado. En comparación con otros anuros de Phyllomedusinae, existen pocos informes sobre el potencial bioactivo de las moléculas derivadas de la piel de esta especie. En esta perspectiva, el objetivo del presente estudio fue evaluar la actividad antibacteriana in vitro de la secreción de la piel de P. azurea mediante la detección de la Concentración Inhibitoria Mínima (CIM) del crecimiento de cepas indicadoras bacterianas y determinar si ocurre un cambio en la permeabilidad de la envoltura celular bacteriana. La determinación de MIC se llevó a cabo mediante el método de la placa de microdilución. La absorbancia se midió y se analizó estadísticamente mediante la prueba t para comparar dos grupos (0.05 de significancia). El impacto del extracto crudo sobre la permeabilidad de la envoltura celular de Staphylococcus aureus ATCC 25923 se realizó mediante el ensayo de cristal violeta, y se midió la absorbancia mediante espectrofotometría seguida del cálculo del porcentaje de absorción de violeta cristal. La CIM específica para S. aureus ATCC 25923 y Escherichia coli ATCC 25922 fue 31.25 μg / ml, mientras que para Bacillus subtilis ATCC 6633 de 125 μg / ml y Pseudomonas aeruginosa ATCC 27853 de 250 μg / ml. El tratamiento con concentraciones crecientes de secreción de piel de rana aumentó la absorción de violeta cristal por las células de S. aureus ATCC 25923, sugiriendo una acción sobre la membrana plasmática de la célula. Los resultados demostraron que la secreción de la piel de P. azurea presenta actividad antibacteriana y amerita más investigaciones para caracterizar las moléculas bioactivas.(AU)

Bryophryne phuyuhampatu sp. n., a new species of Cusco Andes frog from the cloud forest of the eastern slopes of the Peruvian Andes (Amphibia, Anura, Craugastoridae).

A new species of Bryophryne from the humid montane forest of the Department of Cusco, Peru, is described. Specimens were collected at 2795-2850 m a.s.l. in the Área de Conservación Privada Ukumari Llaqta, Quispillomayo valley, in the province of Paucartambo. The new species is readily distinguished from all other species of Bryophryne by having green coloration on dorsum, and blue flecks on flanks and ventral parts. Specimens are characterized by lacking a distinct tympanic annulus, tympanic membrane, and dentigerous processes of vomers, and by having dorsal skin shagreen, discontinuous dorsolateral folds, skin tuberculate on flanks, skin areolate on ventral surfaces of the body, and fingers and toes without lateral fringes or webbing. The new species has a snout-vent length of 14.2-16.9 mm in three males and 22.2-22.6 mm in two females, and is smaller than all other congeneric species except for B. abramalagae. Generic allocation is supported by low genetic distances of the 16S mitochondrial gene and morphological similarity with other species of Bryophryne, and geographic distribution. Bryophryne phuyuhampatusp. n. is only known from the type locality, a cloud forest along the Quispillomayo River in the upper Nusiniscato watershed.

ResumenSe describe una nueva especie de Bryophryne de los bosques nublados del Departamento de Cusco en Perú. Los especímenes fueron colectados a una elevación de 2795­2850 m en el Área de Conservación Privada Ukumari Llaqta, valle del río Quispillomayo, provincia de Paucartambo. La nueva especie se diferencia fácilmente de todas las demás especies de Bryophryne por tener coloración verde en la espalda y manchitas azuladas en los flancos y las superficies ventrales. Los especímenes se caracterizan por carecer de procesos vomerianos y de anillo y membrana timpánicos, y por poseer piel dorsal finamente granulada, pliegues dorsolaterales discontinuos, piel en los flancos verrugosa, y cubierta de verrugas en areola en las partes ventrales, y por carecer de membrana basal y quillas laterales en los dedos. La nueva especie tiene una longitud hocico­cloaca (LHC) de 14.2­16.9 mm en tres machos y de 22.2­22.6 mm en dos hembras, siendo menor que todas las demás especies de Bryophryne excepto por B. abramalagae. Distancias genéticas bajas para el gen mitocondrial 16S, similitud morfológica con otras especies de Bryophryne y distribución geográfica indican que la nueva especie forma parte del género Bryophryne. Bryophryne phuyuhampatusp. n. se conoce únicamente de su localidad tipo, un bosque nublado en el valle del torrente Quispillomayo, en la parte alta de la cuenca del río Nusiniscato.

Effects of past and present livestock grazing on herpetofauna in a landscape-scale experiment.

Livestock grazing is the most widespread land use on Earth and can have negative effects on biodiversity. Yet, many of the mechanisms by which grazing leads to changes in biodiversity remain unresolved. One reason is that conventional grazing studies often target broad treatments rather than specific parameters of grazing (e.g., intensity, duration, and frequency) or fail to account for historical grazing effects. We conducted a landscape-scale replicated grazing experiment (15,000 km2 , 97 sites) to examine the impact of past grazing management and current grazing regimes (intensity, duration, and frequency) on a community of ground-dwelling herpetofauna (39 species). We analyzed community variables (species richness and composition) for all species and built multiseason patch-occupancy models to predict local colonization and extinction for the 7 most abundant species. Past grazing practices did not influence community richness but did affect community composition and patch colonization and extinction for 4 of 7 species. Present grazing parameters did not influence community richness or composition, but 6 of the 7 target species were affected by at least one grazing parameter. Grazing frequency had the most consistent influence, positively affecting 3 of 7 species (increased colonization or decreased extinction). Past grazing practice affected community composition and population dynamics in some species in different ways, which suggests that conservation planners should examine the different grazing histories of an area. Species responded differently to specific current grazing practices; thus, incentive programs that apply a diversity of approaches rather than focusing on a change such as reduced grazing intensity should be considered. Based on our findings, we suggest that determining fine-scale grazing attributes is essential for advancing grazing as a conservation strategy.

A new species of frog of the genus Pristimantis from Tingo María National Park, Huánuco Department, central Peru (Anura, Craugastoridae).

A new species of Craugastoridae frog encountered from 1000-1700 m in elevation in the premontane forests of the Peruvian central Andes is described. The new species is similar in appearance to many other species of Pristimantis, but is easily distinguishable from these species by having bright red coloration on the groin, posterior surface of thighs, and shanks. The new species is only known for two localities 27 km apart in the Huánuco Region.

ResumenDescribimos una nueva especie de rana de la familia Craugastoridae de los bosques premontanos de los Andes centrales peruanos, los especímenes fueron encontrados entre los 1000 ­ 1700 metros de elevación. Esta especie es similar en apariencia a muchas especies de Pristimantis, sin embargo es facilmente distinguible por tener ingles, superficie posterior de los muslos y de la tibia rojo brillante. La nueva especie es conocida solo de dos localidades en la Región Huánuco, ambas separadas por alrededor de 27 km.

External morphological features of the gills of Ichthyophis bannanicus embryo (Amphibia, Gymnophiona) / Características morfológicas externas de las branquias del embrión de Ichthyophis bannanicus (Amphibia, Gymnophiona)

The gills of Ichthyophis bannanicus have yet to be investigated. This paper describes the external morphological features of the gills of mature I. bannanicus embryos exhibiting three pairs of gills on their neck region. Each gill is composed of an axis and filaments. In newly released embryos, the filaments and the axis form at approximately 90° relative to each other; eventually, this angle decreases and the color of the gill fades. The filaments on the axis are arranged alternately, and their spacing varies. The mid-pair of gills is significantly longer by nearly twofold than the front and rear pairs. Likewise, the lengths of the front and rear pairs of gills are not significantly different (P >0.05); for the same pair of gills, the lengths of the left and right parts are not significantly different (P >0.05). The number of filaments is greater in the mid-pair of gills than in the front and rear pairs (P <0.05); the number of filaments in the front pair is not significantly different from that of the rear pair (P >0.05); the number of filaments in the left part does not significantly differ from that of the right parts (P >0.05). Results showed that the gills of I. bannanicus embryo are more similar to those of other species in Ichthyophiidae than to those of species in other families.

Las branquias del Ichthyophis bannanicus aún no se han investigado. En este trabajo se describen las características morfológicas externas de las branquias de embriones maduros de I. bannanicus, que exhiben tres pares de branquias en la región del cuello. Cada branquia está compuesta de un eje y filamentos. En embriones recién liberados los filamentos y la forma del eje es de aproximadamente 90° respecto a la otra; finalmente, este ángulo disminuye y el color de las branquias se desvanece. Los filamentos en el eje están dispuestos en forma alternada y su separación varía. La media de par de branquias es significativamente más larga, por casi el doble que los pares anterior y posterior. Del mismo modo, las longitudes de la parte delantera y posterior de pares de branquias no son significativamente diferentes (p >0,05); para el mismo par de branquias, las longitudes de las partes izquierda y derecha no son significativamente diferentes (p >0,05). El número de filamentos es mayor en el par medio de branquias, que en la parte delantera y posterior de pares (p <0,05); el número de filamentos en el par frontal no es significativamente diferente de la del par trasero (p >0,05); el número de filamentos en la parte izquierda no varía significativamente del de las partes derechas (p >0,05). Los resultados mostraron que las branquias del embrión de I. bannanicus son más similares a los de otras especies en Ichthyophiidae que de otras especies.

Using citizen science data to identify the sensitivity of species to human land use.

Conservation practitioners must contend with an increasing array of threats that affect biodiversity. Citizen scientists can provide timely and expansive information for addressing these threats across large scales, but their data may contain sampling biases. We used randomization procedures to account for possible sampling biases in opportunistically reported citizen science data to identify species' sensitivities to human land use. We analyzed 21,044 records of 143 native reptile and amphibian species reported to the Carolina Herp Atlas from North Carolina and South Carolina between 1 January 1990 and 12 July 2014. Sensitive species significantly associated with natural landscapes were 3.4 times more likely to be legally protected or treated as of conservation concern by state resource agencies than less sensitive species significantly associated with human-dominated landscapes. Many of the species significantly associated with natural landscapes occurred primarily in habitats that had been nearly eradicated or otherwise altered in the Carolinas, including isolated wetlands, longleaf pine savannas, and Appalachian forests. Rare species with few reports were more likely to be associated with natural landscapes and 3.2 times more likely to be legally protected or treated as of conservation concern than species with at least 20 reported occurrences. Our results suggest that opportunistically reported citizen science data can be used to identify sensitive species and that species currently restricted primarily to natural landscapes are likely at greatest risk of decline from future losses of natural habitat. Our approach demonstrates the usefulness of citizen science data in prioritizing conservation and in helping practitioners address species declines and extinctions at large extents.

Quantifying population declines based on presence-only records for red-list assessments.

Accurate trend estimates are necessary for understanding which species are declining and which are most in need of conservation action. Imperfect species detection may result in unreliable trend estimates because this may lead to the overestimation of declines. Because many management decisions are based on population trend estimates, such biases could have severe consequences for conservation policy. We used an occupancy-modeling framework to estimate detectability and calculate nationwide population trends for 14 Swiss amphibian species both accounting for and ignoring imperfect detection. Through the application of International Union for Conservation of Nature Red List criteria to the different trend estimates, we assessed whether ignoring imperfect detection could affect conservation policy. Imperfect detection occurred for all species and detection varied substantially among species, which led to the overestimation of population declines when detectability was ignored. Consequently, accounting for imperfect detection lowered the red-list risk category for 5 of the 14 species assessed. We demonstrate that failing to consider species detectability can have serious consequences for species management and that occupancy modeling provides a flexible framework to account for observation bias and improve assessments of conservation status. A problem inherent to most historical records is that they contain presence-only data from which only relative declines can be estimated. A move toward the routine recording of nonobservation and absence data is essential if conservation practitioners are to move beyond this toward accurate population trend estimation.

Conservation threats and the phylogenetic utility of IUCN Red List rankings in Incilius toads.

Phylogenetic analysis of extinction threat is an emerging tool in the field of conservation. However, there are problems with the methods and data as commonly used. Phylogenetic sampling usually extends to the level of family or genus, but International Union for Conservation of Nature (IUCN) rankings are available only for individual species, and, although different species within a taxonomic group may have the same IUCN rank, the species may have been ranked as such for different reasons. Therefore, IUCN rank may not reflect evolutionary history and thus may not be appropriate for use in a phylogenetic context. To be used appropriately, threat-risk data should reflect the cause of extinction threat rather than the IUCN threat ranking. In a case study of the toad genus Incilius, with phylogenetic sampling at the species level (so that the resolution of the phylogeny matches character data from the IUCN Red List), we analyzed causes of decline and IUCN threat rankings by calculating metrics of phylogenetic signal (such as Fritz and Purvis' D). We also analyzed the extent to which cause of decline and threat ranking overlap by calculating phylogenetic correlation between these 2 types of character data. Incilius species varied greatly in both threat ranking and cause of decline; this variability would be lost at a coarser taxonomic resolution. We found far more phylogenetic signal, likely correlated with evolutionary history, for causes of decline than for IUCN threat ranking. Individual causes of decline and IUCN threat rankings were largely uncorrelated on the phylogeny. Our results demonstrate the importance of character selection and taxonomic resolution when extinction threat is analyzed in a phylogenetic context.

Cost-effective conservation of an endangered frog under uncertainty.

How should managers choose among conservation options when resources are scarce and there is uncertainty regarding the effectiveness of actions? Well-developed tools exist for prioritizing areas for one-time and binary actions (e.g., protect vs. not protect), but methods for prioritizing incremental or ongoing actions (such as habitat creation and maintenance) remain uncommon. We devised an approach that combines metapopulation viability and cost-effectiveness analyses to select among alternative conservation actions while accounting for uncertainty. In our study, cost-effectiveness is the ratio between the benefit of an action and its economic cost, where benefit is the change in metapopulation viability. We applied the approach to the case of the endangered growling grass frog (Litoria raniformis), which is threatened by urban development. We extended a Bayesian model to predict metapopulation viability under 9 urbanization and management scenarios and incorporated the full probability distribution of possible outcomes for each scenario into the cost-effectiveness analysis. This allowed us to discern between cost-effective alternatives that were robust to uncertainty and those with a relatively high risk of failure. We found a relatively high risk of extinction following urbanization if the only action was reservation of core habitat; habitat creation actions performed better than enhancement actions; and cost-effectiveness ranking changed depending on the consideration of uncertainty. Our results suggest that creation and maintenance of wetlands dedicated to L. raniformis is the only cost-effective action likely to result in a sufficiently low risk of extinction. To our knowledge we are the first study to use Bayesian metapopulation viability analysis to explicitly incorporate parametric and demographic uncertainty into a cost-effective evaluation of conservation actions. The approach offers guidance to decision makers aiming to achieve cost-effective conservation under uncertainty.

Differential success in sampling of Atlantic Forest amphibians among different periods of the day / Sucesso diferenciado na amostragem de anfíbios da Mata Atlântica entre diferentes períodos do dia

In general, anurans tend to be nocturnal, though diurnal activity is characteristic of some groups. Studies show that frog activity may be inferred based on the number of individuals collected at different periods of the day, during large-scale field surveys. We investigated the best period of the day to conduct amphibian sampling in nine Atlantic Rainforest areas in southeastern Brazil, based on intensive field surveys. At each locality we employed similar sampling effort during diurnal, crepuscular and nocturnal searches (totaling 704.5 sampling hours). We pooled data from all localities for each period and estimated the proportion of frogs of each species active at each period based on the total number of individuals and on the number of species found during all surveys for that period. We recorded a total of 817 individual frogs from 69 species. Species richness was highest at night (median = 12 species), intermediate at dusk (median = 8), and lowest during the day (median = 4). The percentage of the total number of individual frogs found (pooled species) was highest during the night (ca. 53%) and lowest during the day (ca. 14%). Analyzing each species separately, the number of individuals recorded was consistently higher at dusk and night for most species. Our study evidences a trend for nocturnal activity for most Atlantic Rainforest frogs, with few species having primarily diurnal habits. Those results may favor future studies and conservation efforts for amphibian species.

Em geral, anuros tendem a ser noturnos, apesar de a atividade diurna ser característica de alguns grupos. Estudos mostram que a atividade de anuros pode ser inferida com base no número de indivíduos coletados em diferentes períodos do dia durante pesquisas de campo de larga escala. Nós avaliamos qual o melhor período do dia para conduzir amostragens de anfíbios em nove áreas da Mata Atlântica no sudeste do Brasil, com base em amostragens de campo intensivas. Em cada localidade nós empregamos esforço de amostragem similar durante amostragens diurnas, crepusculares e noturnas (totalizando 704,5 horas de amostragem). Nós agrupamos dados de todas as localidades para cada período e estimamos a proporção de anuros de cada espécie ativa em cada período com base no número total de indivíduos e no número de espécies encontradas durante todas as buscas naquele período. Nós registramos um total de 817 indivíduos pertencentes a 69 espécies. A riqueza de espécies foi maior à noite (mediana = 12 espécies), intermediária ao crepúsculo (mediana = 8) e menor durante o dia (mediana = 4). A porcentagem do número total de indivíduos encontrados (todas as espécies agrupadas) foi maior durante a noite (ca. 53%) e menor durante o período diurno (ca. 14%). Analisando cada espécie separadamente, o número de indivíduos registrados foi consistentemente maior ao crepúsculo e à noite para a maioria das espécies. Nosso estudo evidencia uma tendência para atividade noturna para a maioria dos anuros da Mata Atlântica, com poucas espécies tendo hábitos primariamente diurnos. Esses resultados podem favorecer futuros estudos e esforços de conservação para as espécies de anfíbios.

A new species of Telmatobius (Amphibia, Anura, Telmatobiidae) from the Pacific slopes of the Andes, Peru.

We describe a new species of Telmatobius from the Pacific slopes of the Andes in central Peru. Specimens were collected at 3900 m elevation near Huaytará, Huancavelica, in the upper drainage of the Pisco river. The new species has a snout-vent length of 52.5 ± 1.1 mm (49.3-55.7 mm, n = 6) in adult females, and 48.5 mm in the single adult male. The new species has bright yellow and orange coloration ventrally and is readily distinguished from all other central Peruvian Andean species of Telmatobius but Telmatobiusintermedius by having vomerine teeth but lacking premaxillary and maxillary teeth, and by its slender body shape and long legs. The new species differs from Telmatobiusintermedius by its larger size, flatter head, and the absence of cutaneous keratinized spicules (present even in immature females of Telmatobiusintermedius), and in males by the presence of minute, densely packed nuptial spines on dorsal and medial surfaces of thumbs (large, sparsely packed nuptial spines in Telmatobiusintermedius). The hyper-arid coastal valleys of Peru generally support low species richness, particularly for groups such as aquatic breeding amphibians. The discovery of a new species in this environment, and along a major highway crossing the Andes, shows that much remains to be done to document amphibian diversity in Peru.

ResumenDescribimos una nueva especie de Telmatobius de la vertiente Pacífica de los Andes en el centro de Perú. Los especímenes tipo fueron colectados a una elevación de 3900 m cerca de Huaytará, Huancavelica, en la parte alta de la cuenca del río Pisco. La nueva especie tiene una longitud hocico­cloaca de 52.5 ± 1.1 mm (49.3­55.7 mm, n = 6) en las hembras adultas y de 48.5 mm en un macho adulto. La nueva especie tiene coloración brillante amarilla y naranja en las partes ventrales y se diferencia fácilmente de todas las demás especies peruanas de Telmatobius de los Andes centrales a excepción de Telmatobiusintermedius por tener dientes vomerianos y carecer de dientes maxilares y premaxilares, y por su forma del cuerpo delgada y sus patas largas. La nueva especie se diferencia de Telmatobiusintermedius por su mayor tamaño, por tener la cabeza más plana, y por la ausencia de espículas queratinizadas cutáneas (presentes incluso en hembras inmaduras de Telmatobiusintermedius), y en los machos por la presencia de pequeñas espinas nupciales compactadas en la superficie dorsal y medial de los pulgares (espinas nupciales grandes y dispersas en Telmatobiusintermedius). Los valles costeros hiper-áridos de Perú se caracterizan en general por tener baja riqueza de especies, especialmente para grupos como los anfibios de reproducción acuática. El descubrimiento de una nueva especie en este tipo de ecosistema, y a lo largo de una de las carreteras principales que cruzan los Andes, muestra que aún queda mucho por hacer para documentar la diversidad de los anfibios en Perú.

MORFOHISTOLOGÍA TESTICULAR DE Hypsiboas pulchellus (AMPHIBIA, HYLIDAE)DURANTE LA ESTACIÓN REPRODUCTIVA

En este estudio se aportan datos sobre la morfohistología testicular de Hypsiboas pulchellus durante el período reproductivo con el fin de conocer aspectos de su biología reproductiva. Los testículos, ovoides y amarillentos de 4,64 ± 0,48 mm de largo por 2,05 ± 0,23 mm de ancho, están rodeados por la túnica albugínea que mide 5,60 ± 0,82 µm. Internamente presentan lóculos seminíferos de 257,47 ± 58,25 µm de diámetro, distinguiéndose en ellos cistos con células espermatogénicas asociadas a células de Sertoli. Existe escaso tejido intersticial. Las espermatogonias I son las células más grandes de la serie germinal (14,34 ± 1,74 µm), estas originan a las espermatogonias II (10,14 ± 1,33 µm). Los espermatocitos I presentan la cromatina levemente condensada midiendo 9,34 ± 0,32 µm. Los espermatocitos II miden 8,12 ± 1,07 µm. Las espermátidas I son esféricas, miden 7,61 ± 1,45 µm, y se agrupan en cistos redondeados. Las espermátidas II, en cambio, son alargadas, no hallándoselas dentro de cistos (4,09 ± 0,51 µm). Los espermatozoides, alargados y flagelados, se hallan libres en el centro del lóculo. La morfohistología de los testículos analizados muestra características similares a las observadas en otros anfibios neotropicales, y presentan todas las células del linaje espermatogénico en un mismo lóculo.

In this study, data concerning to the morphological testicular features of Hypsiboas pulchellus during the breeding season is provided, in order to know aspects of their reproductive biology. The testis, ovoid and yellowish with 4.64 ± 0.48 mm length and 2.05 ± 0.23 mm wide, are surrounded by the tunica albuginea which measures 5.60 ± 0.82 µm. The seminiferous locules, placed internally, measure 257.47 ± 58.25 µm in diameter, and cysts with spermatogenic cells associated with Sertoli cells are distinguished in them. The interstitial tissue is scarce. The biggest cells of the germinal series are the spermatogonia I (14.34 ± 1.74 µm), from which spermatogonia II (10.14 ± 1.33 µm) originate. Spermatocytes I measure 9.34 ± 0.32 µm and have slightly condensed chromatin. The spermatocytes II are 8.12 ± 1.07 µm long. The spermatids I are spherical, grouped in rounded cysts, and measure 7.61 ± 1.45 µm. On the other hand, spermatids II are elongated (4.09 ± 0.51 µm) and not within the cysts. Towards the center of the loculus, the free, lengthened and flagellated spermatozoids are located. The morphohistology of the analyzed testicles show similarities with those observed in other neotropical amphibians, being all the cells of the spermatogenic lineage in the same locule.