Acoustic niche partitioning in an anuran community from the municipality of Floriano, Piauí, Brazil / Partição de nicho acústico em uma comunidade de anuros do município de Floriano, Piauí, Brasil

Abstract Vocalizations are an important trait for the identification of cryptic and/or closely related amphibian species. Different vocalizations also contribute to partitioning of the acoustic space by sympatric species. This study aimed to describe the advertisement calls of anurans in a pond of the municipality of Floriano, State of Piauí, Brazil, and infer the acoustic niche partitioning of amphibians. Euclidean distance was used in a cluster analysis approach to infer the acoustic similarities among species. Thirteen species were analysed: Boana raniceps, Dendropsophus nanus, D. rubicundulus , D. minutus, Leptodactylus fuscus, L. troglodytes, L. vastus, Pithecopus nordestinus , Physalaemus cuvieri, P. nattereri, Pleurodema diplolister, Proceratophrys cristiceps and Scinax ruber. From these, six showed more than 90% of acoustic overlap: P. nattereri , P. cuvieri, L. fuscus and L. vastus (Leptodactylidae); and, D. nanus and D. rubicundulus (Hylidae). Despite the acoustic similarities among these six species, the acoustic interference was reduced due to the small number of sympatric species and to distinct features on carrier frequency such as dominant frequency and the degree of modulation in the frequency. Environmental factors limit the periods and sites of reproduction respectively, which may maintain the low anuran diversity and consequently reduce acoustic overlap.

Resumo A vocalização é um componente importante na identificação de espécies crípticas e/ou proximamente relacionadas de anfíbios anuros. Diferenças na bioacústica também contribuem para o particionamento do espaço acústico entre espécies simpátricas. Este estudo descreve o canto de anúncio de anuros em uma lagoa temporária do município de Floriano, Estado do Piauí, Brasil, além de buscar inferir o particionamento do nicho acústico entre essas espécies. Para tanto, foi utilizada a distância Euclidiana para compor uma análise de conglomerados e avaliar a similaridade/dissimilaridade da bioacústica da anurofauna. Treze espécies foram amostradas: Boana raniceps, Dendropsophus nanus, D. rubicundulus , D. minutus, Leptodactylus fuscus, L. troglodytes, L. vastus, Pithecopus nordestinus , Physalaemus cuvieri, P. nattereri, Pleurodema diplolister, Proceratophrys cristiceps e Scinax ruber. Destas, seis apresentaram mais de 90% de sobreposição acústica: P. nattereri, P. cuvieri, L. fuscus e L. vastus (Leptodactylidae); e, D. nanus e D. rubicundulus (Hylidae). Apesar da similaridade acústica entre essas seis espécies, a interferência é reduzida devido ao pequeno número de espécies simpátricas e certas características distintas na frequência do canto, tais como: frequência dominante e sua modulação. Fatores ambientais específicos do semiárido limitam os períodos e sítios reprodutivos, o que mantém baixa diversidade de espécies e consequentemente reduz a sobreposição no nicho acústico.

Anuran diversity in urban and rural zones of the Itacoatiara municipality, central Amazonia, Brazil / Diversidade de anuros nas zonas urbana e rural de Itacoatiara, Amazônia central, Brasil

Few studies in the Amazon region have evaluated anuran diversity in urban forest fragments or in areas with differing degrees of anthropogenic impact. We determined the composition and abundance of the anurofauna in urban and rural sites within the municipality of Itacoatiara in central Amazonia, Brazil. Specimens were sampled from January 2012 to May 2013 in 10 urban sites and five rural sites. A total of 1,538 anurans (930 in rural sites and 608 in urban sites) were recorded, belonging to 29 species in five families. Species richness was higher in rural sites, with 10 exclusive species. All species found in urban sites were also found in rural sites, however, species abundance varied considerably between the habitats. Sampling- and individual-based rarefaction curves showed a tendency toward stabilization of species richness only in the urban sites. We concluded that the anuran assemblages in the urban sites are depauperate due to the absence of many species associated to pristine terra firme or várzea and igapó forests. Habitat loss and quality degradation in urban landscapes are the main factors threatening amphibian diversity. The anuran assemblages in our study areas were similar to those recorded in other Amazonian habitats. Conservation measures involving anurans in this region should consider the preservation of habitat mosaics, including both pristine terra firme, várzea and igapó forests. (AU)

Poucos estudos avaliaram a diversidade de anuros em fragmentos florestais urbanos ou em áreas com diferentes graus de impacto antrópico na Amazônia. Neste estudo foi determinada a composição e abundância de anuros em áreas urbanas e rurais no município de Itacoatiara, na Amazônia central, Brasil. Anuros foram amostrados entre janeiro de 2012 e maio de 2013 em 10 locais na área urbana e cinco na área rural. Um total de 1.538 indivíduos (930 na área rural e 608 na área urbana) foram registrados, compreendendo 29 espécies e cinco famílias. A área rural apresentou maior riqueza de espécies, com 10 espécies exclusivas. Todas as espécies identificadas na área urbana também foram encontradas na área rural, mas a abundância das espécies entre ambas áreas variou consideravelmente. Curvas de rarefação por amostragem e por número de indivíduos tenderam à estabilização na riqueza de espécies somente na área urbana. Concluímos que a comunidade de anuros na área urbana está empobrecida devido à ausência de muitas espécies associadas à ambientes inalterados de florestas de terra firme, várzea e igapó. A degradação da qualidade e a perda de habitat nas paisagens urbanas são os principais fatores que ameaçam a diversidade de anfíbios. A comunidade de anuros registrada neste estudo foi similar às de outros habitats na Amazônia. Medidas de conservação envolvendo anuros nesta região devem considerar a preservação de um mosaico de habitats que inclua áreas naturais de floresta de terra firme, várzea e igapó.(AU)

Deep sequencing analysis of toad Rhinella schneideri skin glands and partial biochemical characterization of its cutaneous secretion

Animal poisons and venoms are sources of biomolecules naturally selected. Rhinella schneideri toads are widespread in the whole Brazilian territory and they have poison glands and mucous gland. Recently, protein from toads' secretion has gaining attention. Frog skin is widely known to present great number of host defense peptides and we hypothesize toads present them as well. In this study, we used a RNA-seq analysis from R. schneideri skin and biochemical tests with the gland secretion to unravel its protein molecules. Methods: Total RNA from the toad skin was extracted using TRizol reagent, sequenced in duplicate using Illumina Hiseq2500 in paired end analysis. The raw reads were trimmed and de novo assembled using Trinity. The resulting sequences were submitted to functional annotation against non-redundant NCBI database and Database of Anuran Defense Peptide. Furthermore, we performed caseinolytic activity test to assess the presence of serine and metalloproteases in skin secretion and it was fractionated by fast liquid protein chromatography using a reverse-phase column. The fractions were partially sequenced by Edman's degradation. Results: We were able to identify several classes of antimicrobial peptides, such as buforins, peroniins and brevinins, as well as PLA2, lectins and galectins, combining protein sequencing and RNA-seq analysis for the first time. In addition, we could isolate a PLA2 from the skin secretion and infer the presence of serine proteases in cutaneous secretion. Conclusions: We identified novel toxins and proteins from R. schneideri mucous glands. Besides, this is a pioneer study that presented the in depth characterization of protein molecules richness from this toad secretion. The results obtained herein showed evidence of novel AMP and enzymes that need to be further explored.(AU)

Acetylcholine produces contractions mediated by the cyclooxygenase pathway in arterial vessels in the Chilean frog (Calyptocephalella gayi) / Acetilcolina produz contrações mediadas pela via da ciclooxigenase em vasos arteriais da rã chileno (Calyptocephalella gayi)

Abstract Previous studies performed in marine fish (I. conceptionis and G. laevifrons) showed that indomethacin blocked arterial contraction mediated by acetylcholine (ACh). The objective of this study was to determine if contraction induced by acetylcholine is mediated by the cyclooxygenase pathway in several arterial vessels in the Chilean frog Calyptocephalella gayi. Arteries from the pulmonary (PA), dorsal (DA), mesenteric (MA) and iliac (IA) regions were dissected from 6 adult specimens, and isometric tension studies were done using dose response curves (DRC) for ACh (10-13 to 10-3 M) in presence of a muscarinic antagonist (Atropine 10-5 M) and an unspecific inhibitor of cyclooxygenases (Indomethacin, 10-5M). All the studied arteries exhibited vasoconstriction mediated by ACh. This vasoconstriction was abolished in the presence of atropine in DA, MA and IA and attenuated in PA. Indomethacin abolished the vasoconstriction in MA and attenuated the response in PA, DA and IA. Similar to marine fish, C. gayi have an ACh-mediated vasoconstrictor pattern regulated by muscarinic receptors that activate a cyclooxygenase contraction pathway. These results suggest that the maintenance in vasoconstrictor mechanisms mediated by ACh→COX →vasoconstriction is conserved from fish to frogs.

Resumo Estudos feitos em peixes marinhos (I. conceptionis e G. laevifrons) têm demostrado que a indometacina bloqueia a contração arterial mediada por acetilcolina (ACh). O objetivo do presente estudo foi avaliar o efeito da via da ciclooxigenase na contração induzida por ACh em vasos arteriais da rã chilena Calyptocephalella gayi. Foram dissecadas regiões das artérias pulmonares (PA), dorsal (DA), mesentérica (MA) e ilíaca (IA) de seis espécimes adultos e realizados estudos de tensão isométrica utilizando curvas dose-resposta (CDR) de ACh (10-13 a 10-3 M) na presença de um antagonista muscarínico (atropina, 10-5 M) e um inibidor das ciclooxigenases (indometacina, 10-5 M). Todas as artérias evidenciaram uma resposta vasoconstritora mediada por ACh. Esta resposta vasoconstrictora foi suprimida na presença de atropina nas artérias DA, MA, IA e atenuada na PA. A indometacina suprimiu a vasoconstrição na artéria MA e atenuou a resposta nas artérias PA, DA e IA. Tal como os peixes marinhos, a C. gayi tem um padrão de vasoconstrição mediado por Ach que é regulado pelos receptores muscarínicos e pela ciclooxigenase. Estes resultados sugerem a conservação dos mecanismos vasoconstrictores mediados por ACh→COX em peixes e rãs.

Correlations of condition factor and gonadosomatic, hepatosomatic and lipo-somatic relations of Leptodactylus macrosternum (ANURA: Leptodactylidae) in the Brazilian Semi-arid

ABSTRACT The objective of this study was to assess variations of the condition factor (K1) in relation to the gonadosomatic- RGS and energy reserves (hepatosomatic - RWL and liposomatic - RFB relations) of Leptodactylus macrosternum and their relationship to climate variation in the Northeast of Brazil, Caatinga area, state of Paraiba. The animals were captured fortnightly through active collecting, between January and December 2013. Significant differences were observed in the monthly variations of K1, RGS and RFB indices in male and female L. macrosternum over the months of collection. In males, K1 showed no significant relationship with the other variables. In females, RGS values only show notable correlations with RWF and K1 values. K1 values showed significant correlations with all other weight and length ratios. Climate change in the HFOB region showed significant relationships with the variation of the indexes evaluated, with the exception of RWF. The variation of K1, RGS, RWL and RFB values over the months of collection as well as their relation with the local climatic variation, showed a brief reproductive activity for the species.

First record of the Lesser Snouted Treefrog Scinax nasicus (Cope, 1862) in Brazilian coast and new species records for the state of Rio Grande do Sul / Primeiro registro da perereca-de-banheiro Scinax nasicus (Cope, 1862) na costa do Brasil e novos registros para o estado do Rio Grande do Sul

Abstract Herein, we provide new occurrence records of Scinax nasicus (Cope, 1862) for the state of Rio Grande do Sul, Southern Brazil. All new records here provide are located on Southern half of the state. Besides this, we provide the first record for species in Brazilian coastal zone. Those records improve considerably our knowledge regarding species distribution in Southern Brazil.

Resumo Aqui, nós fornecemos novos registros de ocorrência de Scinax nasicus (Cope, 1862) para o estado do Rio Grande do Sul, Sul do Brasil. Todos os novos registros aqui fornecidos estão localizados na metade sul do estado. Além disso, nós fornecemos o primeiro registro para a espécie na zona costeira brasileira. Esses registros melhoram consideravelmente o nosso conhecimento sobre a distribuição da espécie no Sul do Brasil.

The peculiar breeding biology of the Amazonian frog Allobates subfolionidificans (Aromobatidae)

ABSTRACT Allobates subfolionidificans is a vulnerable and endemic leaf-litter frog from the state of Acre, Brazilian Amazonia. We monitored a population of A. subfolionidificans through regular censuses and mark-recapture of 181 individuals during an entire breeding season to characterize its reproductive behavior. The space use of A. subfolionidificans individuals differed between sexes, with males using smaller and more segregated spaces. Males defended territories and were aggressive against same-sex individuals, which was not the case in females. The daily cycle of calling activity showed peaks in the morning and in the afternoon, and the occurrence of reproductive events was positively correlated with monthly rainfall. The breeding behavior comprised vocal and tactile interactions, although the species lacked reproductive amplexus. Egg and larvae attendance, as well as tadpole transport to water environments was performed mostly by males but occasionally by females, probably in cases of desertion by the father. This species is characterized by performing courtship, mating, oviposition, as well as egg and larvae attendance exclusively on the under surface of leaves, a unique behavior among members of the superfamily Dendrobatoidea.

Ocellatin peptides from the skin secretion of the South American frog Leptodactylus labyrinthicus (Leptodactylidae): characterization, antimicrobial activities and membrane interactions

Background The availability of antimicrobial peptides from several different natural sources has opened an avenue for the discovery of new biologically active molecules. To the best of our knowledge, only two peptides isolated from the frog Leptodactylus labyrinthicus, namely pentadactylin and ocellatin-F1, have shown antimicrobial activities. Therefore, in order to explore the antimicrobial potential of this species, we have investigated the biological activities and membrane interactions of three peptides isolated from the anuran skin secretion. Methods Three peptide primary structures were determined by automated Edman degradation. These sequences were prepared by solid-phase synthesis and submitted to activity assays against gram-positive and gram-negative bacteria and against two fungal strains. The hemolytic properties of the peptides were also investigated in assays with rabbit blood erythrocytes. The conformational preferences of the peptides and their membrane interactions have been investigated by circular dichroism spectroscopy and liposome dye release assays. Results The amino acid compositions of three ocellatins were determined and the sequences exhibit 100% homology for the first 22 residues (ocellatin-LB1 sequence). Ocellatin-LB2 carries an extra Asn residue and ocellatin-F1 extra Asn-Lys-Leu residues at C-terminus. Ocellatin-F1 presents a stronger antibiotic potential and a broader spectrum of activities compared to the other peptides. The membrane interactions and pore formation capacities of the peptides correlate directly with their antimicrobial activities, i.e., ocellatin-F1 > ocellatin-LB1 > ocellatin-LB2. All peptides acquire high helical contents in membrane environments. However, ocellatin-F1 shows in average stronger helical propensities. Conclusions The obtained results indicate that the three extra amino acid residues at the ocellatin-F1 C-terminus play an important role in promoting stronger peptide-membrane interactions and antimicrobial properties. The extra Asn-23 residue present in ocellatin-LB2 sequence seems to decrease its antimicrobial potential and the strength of the peptide-membrane interactions.(AU)

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.

Living in a same microhabitat should means eating the same food? Diet and trophic niche of sympatric leaf-litter frogs Ischnocnema henselii and Adenomera marmorata in a forest of Southern Brazil / Viver no mesmo ambiente significa comer o mesmo alimento? Dieta e nicho trófico dos anuros simpátricos de folhiço Ischnocnema henselii e Adenomera marmorata em uma floresta do sul do Brasil

In this study we analyzed diet composition, niche breadth and overlap of the two leaf-litter frogs Ischnocnema henselii and Adenomera marmorata. Frogs were collected in an Atlantic Rainforest area in the Reserva Natural Salto Morato, in Paraná State, Southern Brazil, using plots of 16 m2 established on forest floor. Ischnocnema henselii consumed 18 different types of prey and the diet of this species was composed predominantly by Hymenoptera (Formicidae) (15.4%), Araneae (13.83%), Orthoptera (6.15%) and Opiliones (6.15%), whereas Adenomera marmorata consumed 15 different types of prey and its diet was composed mainly by Hymenoptera (Formicidae) (45.7%), Acari (31.8%) and Blattodea (14.8%). The niche breadth of I. henselii was BA = 0.43 and that of A. marmorata was BA = 0.19. The diet of the two sympatric species of leaf-litter frogs was basically composed by arthropods and the trophic niche overlap among them did not differ from expected at random. The differences in prey consumption should potentially facilitate the coexistence of two sympatric frogs on the forest floor. Possibly, this difference of prey consumption partly reflects differences in jaw width, species-specific body size of the two species and the period of activity of these two species.

Neste estudo analisamos a composição da dieta e a amplitude e sobreposição do nicho trófico dos anuros de folhiço Ischnocnema henselii e Adenomera marmorata. Os anuros foram coletados em uma área de Mata Atlântica na Reserva Natural Salto Morato, no Estado do Paraná, Sul do Brasil, utilizando o método de parcelas de 16 m2 estabelecidas no chão da floresta. Ischnocnema henselii consumiu 18 diferentes tipos de presa e a dieta desta espécie foi composta predominantemente por Hymenoptera (Formicidae) (15,4%), Araneae (13,83%), Orthoptera (6,15%) e Opiliones (6,15%), enquanto Adenomera marmorata consumiu 15 diferentes tipos de presas e sua dieta foi composta principalmente por Hymenoptera (Formicidae) (45,7%), Acari (31,8%) e Blattodea (14,8%). A amplitude de nicho de I. henselii foi BA = 0,43 e de A.marmorata foi BA = 0,19. A dieta das duas espécies de anuros simpátricos do folhiço por nós estudadas foi composta basicamente de artrópodes e a sobreposição de nicho trófico entre elas não diferiu do esperado para ocorrer ao acaso. As diferenças no consumo de presas potencialmente devem facilitar a coexistência desses dois anuros simpátricos no chão da floresta. Possivelmente, esta diferença no consumo de presas em parte reflete diferenças na largura da mandíbula, no tamanho do corpo e no período de atividade dessas duas espécies.

The recognition of Dendropsophus minusculus () (Hylidae, Dendropsophini) as a highly polymorphic, multi-domain distributed species / O reconhecimento de Dendropsophus minusculus () (Hylidae, Dendropsophini) como espécie polimórfica e de ocorrência em distintos domínios morfoclimáticos

Herein we present new occurrences of sites of Dendropsophus minusculus in Brazil adding information about reproductive biology and colour patterns of the species. Such information is fundamental for characterising the species that we believe has been neglected due to its morphological similarity with other congeneric species. Dendropsophus minusculus may be found in different morphoclimatic domains, being one of the most generalist species of the genus in this aspect. The species plasticity is not restricted to its occurrence, but also related to aspects of its reproductive biology, and we hypothesised that the latter feature could explain the wide geographical range of the species. We highlight the importance of further in-depth studies and the use of D. minusculus as a model to understand the historical events responsible for the current geographical distribution of the morphoclimatic domains in Brazil.

No presente estudo relatamos novos locais de ocorrência para Dendropsophus minusculus no Brasil e acrescentamos informações sobre a biologia reprodutiva e padrões de coloração da espécie. Essas informações são de fundamental importância para a caracterização da espécie, que acreditamos ter sido negligênciada em virtude de sua semelhança morfológica com outras espécies congenéricas. Dendropsophus minusculus pode ser encontrada em diversos domínios morfoclimáticos, sendo, neste aspecto, uma das espécies mais generalistas de seu gênero. A plasticidade da espécie não se restringe a sua ocupação territorial, mas também a aspectos relativos a sua biologia reprodutiva e especulamos que esta seja a razão de sua amplitude territorial. Destacamos ainda a importância de estudos mais aprofundados e o uso da espécie como modelo para o entendimento de eventos pretéritos responsáveis pela distribuição de domínios morfoclimáticos no Brasil.

Reproductive biology of Pleurodema guayapae (Anura: Leptodactylidae: Leiuperinae) / Biología reproductiva de Pleurodema guayapae (Anura: Leptodactylidae: Leiuperinae)

Pleurodema guayapae is a species that inhabits saline environments and semidesert zones from central Argentina. To date, the knowledge about the reproductive biology of this species is very poor, and our aim is to contribute to its knowledge with the description of some important reproductive aspects. For this, field work was undertaken in an area near to Patquía, La Rioja province. Sampling was undertaken during three summer periods (2006-2007; 2007-2008; 2008-2009) in Chamical-Patquía area, where we could find reproductively active populations. We observed and described breeding sites, type of clutch, process of foam nest construction, clutch and egg number and sizes, and hatching time and stage. Behaviour observations were performed from the time that males began to call until the pairs ended up the foam nests building, and layed the eggs. Additionally, one amplected pair was observed and filmed in the process of foam nest construction, and four amplectant pairs were collected and separatelly placed in plastic containers, for nests observations in the laboratory. Hatching time was based on three different foam nests of known age. We found that P. guayapae populations were acoustically active only after a rainfall. Its breeding sites were represented by ephemeral ponds of fresh water, product of rains. The males emitted their calls inside or outside these ponds. A detailed description of the foam nest construction process by both females and males was made. The clutches were in dome-shaped foam nest type of 6-9cm in diameter and 1-3cm in height, some of which were in communal nests. The nests had an average of 1 137 pigmented eggs. This species showed a short hatching time. Our results allow us to conclude that this species should be considered an extreme explosive breeder. Our results are discussed with others obtained for related species.

Pleurodema guayapae es una especie que habita en zonas semidesérticas y salinas del área central de Argentina. Los conocimientos actuales sobre la biología reproductiva de esta especie son muy pobres, por lo que nuestro objetivo fue contribuir a su conocimiento con la descripción de algunos aspectos reproductivos importantes. Para ello se realizaron tres campañas de muestreo durante los veranos (2006-2007, 2007-2008 y 2008-2009) en el área de Chamical-Patquía, provincia de La Rioja, donde se encontraron poblaciones reproductivamente activas. Se observaron y describieron los sitios reproductivos, tipo de puesta, proceso de construcción del nido de espuma, tamaño de la puesta y de los huevos, número de huevos por puesta y tiempo y estado de eclosión de la larva. Las observaciones de comportamiento se realizaron desde el momento en que los machos comenzaron a emitir los cantos hasta que la pareja en amplexo terminó de construir el nido de espuma con los huevos en su interior. Además, se observó y filmó una pareja en amplexo, en el proceso de construcción del nido de espuma y se recolectaron cuatro parejas en amplexo y se colocaron separadamente en recipientes plásticos para que sus puestas pudieran ser observadas en el laboratorio. El tiempo de eclosión se determinó en tres nidos de espuma de edad conocida. Se encontró que las poblaciones de P. guayapae solo estuvieron acústicamente activas luego de una lluvia torrencial. Los sitios de reproducción fueron charcas efímeras de agua de lluvia. Los machos emitieron sus cantos de advertencia desde adentro o fuera de esas charcas. Se realizó una descripción detallada del proceso de construcción del nido de espuma por parte de la hembra y el macho de P. guayapae. El tipo de la puesta en esta especie es un nido de espuma flotante con forma de domo de 6 a 9cm de diámetro y de 1 a 3cm de alto, algunos de los cuales estaban unidos entre sí formando puestas comunales. Los nidos de espuma poseen en promedio 1 137 huevos pigmentados. Esta especie mostró un tiempo de eclosión corto. Nuestros resultados nos permiten concluir que P. guayapae debe ser considerada como reproductor explosivo extremo. Nuestros resultados son discutidos con los obtenidos en otras especies afines.

Ecología reproductiva de Geobatrachus walkeri (Anura: Srabomantidae) / Reproductive ecology of Geobatrachus walkeri (Anura: Strabomantidae)

Geobatrachus walkeri is a frog that belongs to a monotypic genus and is endemic to the Sierra Nevada de Santa Marta, Colombia. The species inhabits different microhabitats between 2 000 to 3 500m altitude, including the leaf litter of a pine plantation. To understand its reproductive ecology, we conducted eight frog samplings, covering the rainy and dry seasons, and two habitat types (secondary native forest and pine plantation) during 2010-2011. For this study, we also included data obtained from five previous similar samplings undertaken during 2008-2009. The pine leaf litter was the main microhabitat where frogs were found; we heard choruses of six-ten calling males during all sampled months, and observed the frogs having diurnal and nocturnal activity. Regardless of the year of study, the population consisted of neonates recruited several times of the year, a large number of juveniles with a wide range of body sizes, and fewer adults with a narrower range of body size. The histological analyses of the gonads showed that the size at maturity was near 18mm SVL for males and females, and those adult males and females were reproductive active during all sampling months, suggesting a continuous reproductive activity pattern. However, during the dry season, the seminiferous tubules showed a drastically diminished spermatic epithelium although containing abundant luminal spermatozoa, which suggest a reduction in the sperm production at the end of this season. Similarly, frogs of all age categories were significantly more abundant during the early dry season, whereas were significantly less abundant with the advancement of the dryness suggesting that the intensity of the dry season could temporally stop the activity and reproduction of this population.

Geobatrachus walkeri es una rana que pertenece a un género monotípico y es endémica de la Sierra Nevada de Santa Marta, Colombia. La especie habita en diferentes microhábitats entre los 2 000 y 3 500m de altitud, incluyendo la hojarasca de una plantación de pinos. Para entender su ecología reproductiva se realizaron ocho muestreos de individuos, que abarcaron las estaciones de lluvias y de sequía y dos tipos de hábitats (bosque secundario y plantación de pino) durante 2010-2011. También se incluyeron los datos de cinco muestreos similares obtenidos previamente durante el período 2008-2009. El principal microhábitat donde las ranas se encontraron fue la hoja-rasca de pino, allí escuchamos coros de seis-diez machos durante todos los meses muestreados, y observamos que las ranas tienen actividad diurna y nocturna. Sin importar el año de estudio, la población se compone de neonatos reclutados en varios meses del año, un gran número de juveniles con un amplio rango de tamaños corporales, y un menor número de adultos con un rango más reducido de tamaño corporal. Los análisis histológicos de las gónadas mostraron que el tamaño a la madurez se adquiere cerca de los 18mm LRC para machos y hembras, y que los machos y hembras adultos fueron reproductivos durante todo el muestreo, lo que sugiere un patrón continuo de actividad reproductiva. Sin embargo, durante la estación seca, los túbulos seminíferos presentan un epitelio espermático drásticamente disminuido, aunque contienen abundantes espermatozoides luminales, lo que sugiere una reducción en la producción de espermatozoides en el final de la estación seca. Del mismo modo, las ranas de todas las categorías de edad fueron significativamente más abundantes durante el inicio de la estación seca, mientras que fueron significativamente menos abundantes con el avance de la sequedad, lo que sugiere que una mayor intensidad de la temporada de sequía podría detener temporalmente la actividad y la reproducción en esta población.

Demografía poblacional de Rhinella arenarum (Anura: Bufonidae) y Physalaemus biligonigerus (Anura: Leiuperidae) en agroecosistemas de la provincia de Córdoba, Argentina / Population demography in Rhinella arenarum (Anura: Bufonidae) and Physalaemus biligonigerus (Anura: Leiuperidae) in agroecosystems in the province of Córdoba, Argentina

The advancing agricultural frontier has led to an important loss of natural habitats, with significant consequences for biodiversity. The demography for two species of anurans, Physalaemus biligonigerus and Rhinella arenarum, both associated with agricultural systems in the central region of the Córdoba Province, were analyzed and compared in this study. Four sites were sampled: three agroecosystems with different alteration degrees (C1, C2 and SM1) and a fourth site not cultivated (SM2). The sampling was conducted during two reproductive periods, from September 2008 to April 2009 and September 2009 to April 2010. Individuals were captured using live pitfall traps for the metamorphic, juveniles and adults; and visual encounter survey, for the capture of eggs and larvae. With the abundance data, the survival for each age class was estimated using the KNM method (Kiritani Nakasuki Manly). With survival rates and fertility population, Leslie matrices were elaborated to obtain a quantitative projection of the population size. Altered environments showed lower eggs and larvae survival. Population pro-jections were favorable in the site SM2 and were less favorable and a tendency to extinction, in sites dominated by crops. This study showed that the agroecosystems of this region are possibly inhospitable environments for reproduction and survival of the species studied. The aquatic stages in the life cycle of both species would be the more affected, since water bodies deterioration is present or may occur in those areas. We can recognize species-specific effects of agricultural ecosystems; P. biligonigerus was the most affected species, possibly because of their life histories and habitat requirements. We suggested that environmental degradation caused by the cropland in the central region of Argentina would impact on the demographics of the anuran populations in the area.

El crecimiento de la agricultura produce pérdida del hábitat natural, con consecuencias para la biodiversidad de los anfibios. Se analizó la demografía poblacional de dos especies de anfibios en agroecosistemas de la provincia de Córdoba, Rhinella arenarum y Physalaemus biligonigerus. Los muestreos se realizaron en tres agroecosistemas y en un cuarto sitio, no afectado por cultivos. Los individuos se capturaron utilizando trampas de caída viva y relevamientos visuales. Con los datos de abundancia se estimó la supervivencia para cada clase de edad y con las tasas de supervivencia y fertilidad poblacional, se construyeron matrices de Leslie para obtener una proyección cuantitativa del tamaño poblacional. Las proyecciones poblacionales fueron favorables para el sitio SM2 y desfavorables para los sitios cultivados. Los agroecosistemas son ambientes posiblemente inhóspitos para la reproducción y supervivencia de las especies estudiadas. Particularmente se ven afectados aquellos estadios acuáticos del ciclo de vida de estas especies, debido al deterioro de los cuerpos de agua en estas áreas. Se reconocieron efectos especie-específicos de los ecosistemas agrícolas, ya que P. biligonigerus resultó la especie más afectada. Se sugiere un deterioro ambiental producido por cultivos extensivos de soja, lo que impactaría la demografía de las poblaciones de anuros.

Habitat selection by anurofauna community at rocky seashore in coastal Atlantic Forest, Southeastern Brazil / Selecao de habitat por uma comunidade de anuros em um costao rochoso na Mata Atlantica costeira, sudeste do Brasil

Rocky seashores are low granitic hills distributed along the southeastern Brazilian coast with xeric-like vegetation due to the shallow soil. Knowledge on amphibian communities and their reproductive patterns is especially reduced on this kind of environment. Herein, we present a framework of two years monitoring an amphibian community at a rocky seashore environment located at the protected area of Parque Estadual da Serra da Tiririca, municipality of Niterói, state of Rio de Janeiro, Brazil. We conducted diurnal and nocturnal searches for frogs in tank bromeliads, rocky surface and shrubby vegetation. Annual pattern of breeding activity of anurans was also estimated. Individuals of the most abundant tank-bromeliad, Alcantarea glaziouana were collected and measured according to several variables to understanding the selection of bromeliads by frogs. We checked the influence of the environmental conditions on amphibian abundance, association between the bromeliads measures, and the water storage in the tank. We recorded the species: Scinax aff. x-signatus; S. cuspidatus; S. littoreus; Thoropa miliaris and Gastrotheca sp. Bromeliads were the preferential habitat used by anurans. The nocturnal habit was predominant for all species and during diurnal searches, the specimens were found sheltered in bromeliads axils. The number of calling males as well as amphibian abundance was associated with the rainiest and warmest period of the year. The species S. littoreus was observed in breeding activity in the majority of sample period. Adult calling males of T. miliaris were observed especially in the rainy season. Rainfall and temperature combined are positively correlated to the total number of captured amphibians. However, individually, rainfall was not significantly correlated, while temperature was positively correlated with the amphibian abundance. Water storage capacity by bromeliads was correlated to characteristics and size of the plant. In the rainy season, the height of the plant and the diameter on top view were correlated with the occurrence of amphibians, while during the driest period there was no correlation among variables and the bromeliad usage by amphibians. Recorded species were strongly associated to the Atlantic Forest domain. Nevertheless, the occupation of rocky seashores by anurans may be more associated with the specialized reproductive modes presented by species, since there is no permanent water available in ponds or streams.

Costões rochosos são morros graníticos distribuídos ao longo da costa do sudeste brasileiro, com vegetação xeromórfica devido ao seu solo raso. O conhecimento sobre comunidades de anfíbios e seus padrões reprodutivos é especialmente reduzido neste tipo de ambiente. Neste estudo, nós apresentamos os resultados de dois anos de monitoramento de uma comunidade de anfíbios em um costão rochoso localizado na área protegida do Parque Estadual da Serra da Tiririca, município de Niterói, estado do Rio de Janeiro, Brasil. Nós fizemos buscas diurnas e noturnas para encontrar anuros em bromélias-tanque, superfície rochosa e vegetação arbustiva. O padrão anual de atividade reprodutiva também foi estimado. Indivíduos da bromélia-tanque mais abundante no local, Alcantarea glaziouana, foram coletados e tiveram diversas variáveis mensuradas a fim de se entender a seleção de bromélias por anuros. Verificamos a influência das condições ambientais sobre a abundância dos anfíbios, e a associação entre as medidas das bromélias e a capacidade de estocar água no seu tanque. Diferenças na utilização de microhábitats e na ocupação de bromélias pelos anuros também foram investigadas, assim como também foi testada a influência das variáveis das bromélias sobre a ocorrência dos anuros nestas plantas. Foram registradas as seguintes espécies: Scinax aff. x-signatus; S. cuspidatus; S. littoreus; Thoropa miliaris e Gastrotheca sp. As bromélias foram o habitat preferido pelos anuros. O hábito noturno foi predominante para todas as espécies. Durante buscas diurnas, os espécimes foram encontrados abrigados nas axilas das folhas das bromélias. O número de machos vocalizando, assim como a abundância de anfíbios, foi associado com o período mais quente e chuvoso. Machos adultos de T. miliaris foram observados vocalizando especialmente na estação chuvosa. A precipitação e a temperatura combinadas foram positivamente correlacionadas com o número total de anfíbios capturados. Entretanto, individualmente, a precipitação não foi significativamente correlacionada, enquanto que a temperatura foi positivamente correlacionada com a abundância de anfíbios. A capacidade de armazenamento de água pelas bromélias está correlacionada com as características e tamanho da planta. Na estação chuvosa, a altura da planta e o seu diâmetro foram correlacionados com a ocorrência de anfíbios, enquanto que durante o período seco não existiu correlação entre variáveis e o uso da bromélia pelos anfíbios. As espécies registradas estão fortemente associadas ao domínio da Floresta Atlântica. Entretanto, a ocupação dos costões rochosos por anuros pode estar mais associado com modos reprodutivos especializados apresentados pelas espécies, já que não existem riachos ou poças d'água permanentes disponíveis no local.

Alimentación de dos especies de anfibios (Anura: Hylidae) en la estación de bajas temperaturas y su relación con la acumulación de energía en Santa Fe, Argentina / Feeding of two amphibian species (Anura: Hylidae) during the low temperatures season and its relationship with energy storage in Santa Fe, Argentina

In environments with thermal and pluvial seasonality such as those of the Middle Paraná River floodplain (Province of Santa Fe, Argentina), most amphibian species reproduce during the warm season and drastically diminish their activity during winter. Even though, a few species remain active during the cold season, such as Hypsiboas pulchellus that has its reproductive peak during the autumn-winter period (and the consequent energy demand). The objective of this study was to analyze and compare the feeding and development of fat bodies during the low temperature season for H. pulchellus and Dendropsophus nanus. We analyzed entire gastrointestinal tract contents of both species (H. pulchellus=110 specimens; D. nanus=114 specimens) and applied an index (IRI%) that combines prey abundance, volume and frequency to describe frogs diets; we used fat bodies weights as indicators of stored energy reserves. We compared diet between species with a niche overlap index (Ojk: 0-1) and used null models to ascribe statistical significance to evaluate overlap; and we analyzed variation in empty guts proportions through months and between species. Also, using ANCOVAs we explored differences in fat bodies, number and volume of prey consumed along months, between species and sexes. The most important preys in H. pulchellus diet during the cold season were Araneae (IRI%=34.96), Chironomidae (IRI%=33.08), Tipulidae (IRI%=11.44) and Gryllidae (IRI%=7.31); while for D. nanus, Chironomidae (IRI%=48.14), Tipulidae (IRI%=18.41), Psychodidae (IRI%=7.44) and Araneae (IRI%=7.34). Diet overlap between species was elevated (Ojk=0.78) and higher than expected by chance (mean simulated indices: Ojk=0.04; p[observed≥expected]<0.01; p[observed≤expected]=1). In H. pulchellus there was a monthly variation in number of prey per gut, while in D. nanus there was a difference in fat bodies development between sexes. Fat bodies development, number of prey per gut and preys volume also varied between species. Despite diet similarity between H. pulchellus and D. nanus, each species showed a different strategy to accumulate energy and support their activity during the cold season. The low rate of gastrointestinal emptiness in H. pulchellus (<10% in any analyzed month) together with the poor development of their fat bodies, allows us to point out that, to sustain the breeding elevated energy demands, this species continues with a high feeding rate even at the low temperatures of cold season. On the other hand, the rate of gastrointestinal emptiness of D. nanus was higher than that of H. pulchellus (May=17.24%, July=22.22% and August=35.71%), while their fat bodies were well developed. Thus, D. nanus would depend more on their stored reserves to sustain the energy demands of being active during the low temperatures season and hence would reach the reproductive season in spring-summer in good body condition.

Existe escasa información sobre la alimentación de las especies de anfibios activas durante la estación fría o las estrategias de utilización de reservas lipídicas como fuente de energía durante el periodo de bajas temperaturas. Por lo tanto, se analizaron las estrategias de obtención y acumulación energética durante la estación de bajas temperaturas en un anfibio que se reproduce principalmente en otoño-invierno, Hypsiboas pulchellus (Hp), y uno que lo hace en primavera-verano, Dendropsophus nanus (Dn). Para ello se analizó la alimentación de ambas especies y se utilizó el peso de los cuerpos grasos (CG) como indicador del desarrollo de las reservas energéticas. Si bien se encontró una elevada similitud en la dieta entre las especies, esto no implicó la ausencia de diferencias en las estrategias de obtención energética entre ellas. El bajo índice de vacuidad gastrointestinal de Hp, acompañado de CG poco desarrollados, permite señalar que, para sostener la elevada demanda energética de la actividad reproductiva concentrada en el otoño-invierno, esta especie continúa alimentándose aún en temperaturas bajas. Por otro lado, el índice de vacuidad de Dn fue mayor al de Hp, mientras que sus CG se encontraron bien desarrollados, por lo que esta especie utilizaría en mayor medida sus reservas lipídicas como fuente de energía para transitar el período de bajas temperaturas y llegar en buenas condiciones a la temporada reproductiva en la próxima primavera-verano.

Bioacústica del canto de advertencia de Ceratophrys cranwelli (Anura: Ceratophrydae) / Bioacoustic of the advertisement call of Ceratophrys cranwelli (Anura: Ceratophryidae)

The advertisement call plays an important role in the life history of anuran amphibians, mainly during the breeding season. Call features represent an important character to discriminate species, and sound emissions are very effective to assure or reinforce genetic incompatibility, especially in the case of sibling species. Since frogs are ectotherms, acoustic properties of their calls will vary with temperature. In this study, we described the advertisement call of C. cranwelli, quantifying the temperature effect on its components. The acoustic emissions were recorded during 2007 using a DAT record Sony™ TCD-100 with stereo microphone ECM-MS907 Sony™ and tape TDK™ DAT-RGX 60. As males emit their calls floating in temporary ponds, water temperatures were registered after recording the advertisement calls with a digital thermometer TES 1 300±0.1°C. Altogether, 54 calls from 18 males were analyzed. The temporal variables of each advertisement call were measured using oscillograms and sonograms and the analyses of dominant frequency were performed using a spectrogram. Multiple correlation analysis was used to identify the temperature-dependent acoustic variables and the temperature effect on these variables was quantified using linear regression models. The advertisement call of C. cranwelli consists of a single pulse group. Call duration, Pulse duration and Pulse interval decreased with the temperature, whereas the Pulse rate increased with temperature. The temperature-dependent variables were standardized at 25°C according to the linear regression model obtained. The acoustic variables that were correlated with the temperature are the variables which emissions depend on laryngeal muscles and the temperature constraints the contractile properties of muscles. Our results indicated that temperature explains an important fraction of the variability in some acoustic variables (79% in the Pulse rate), and demonstrated the importance of considering the effect of temperature in acoustic components. The results suggest that acoustic variables show geographic variation to compare data with previous works.

El canto de advertencia en anuros es especie-específico, pero como los anfibios son organismos ectotermos, las variables acústicas pueden estar afectadas por la temperatura. Se describe el canto de advertencia de Ceratophrys cranwelli cuantificando el efecto de la temperatura sobre sus componentes. Los cantos se grabaron in situ y se registró la temperatura del sitio de canto. Un total de 54 cantos de 18 individuos fueron digitalizados y analizados. Las variables temporales de cada canto de advertencia fueron medidas a partir de los oscilogramas y sonogramas, mientras que el análisis de frecuencia se realizó a través del espectrograma. Mediante análisis de correlación múltiple se identificaron las variables acústicas temperatura-dependientes y el efecto de la temperatura fue cuantificado mediante los modelos de regresión lineal obtenidos. C. cranwelli posee un canto de advertencia simple pulsado. Las variables Duración del canto, Duración del pulso e Intervalo entre pulso decrecen con la temperatura, en tanto que la Tasa de pulsos está directamente correlacionada con la temperatura. Las variables temperatura-dependientes fueron estandarizadas a 25ºC según el modelo de regresión lineal obtenido para ser comparadas con datos publicados. Se demuestra la importante variabilidad que produce la temperatura en algunas propiedades del canto de C. cranwelli.

Seasonal testicular changes in Dendropsophus minutus Peters, 1872 (Anura, Hylidae)

The reproductive cycle in anurans may be either continuous or discontinuous. These differences may be connected to seasonal climate changes and/or to anthropic activity. Forty adult male individuals of the Dendropsophus minutus species were collected during one year, in the municipality of Chapada dos Guimarães (Mato Grosso, Brazil). The testicles were studied under light and transmission electron microscopy. No variations were observed when the diameter of the seminiferous tubules and the thickness of the interstitial tissue were studied. However, changes in spermatogenesis were conspicuous and indicated that the reproductive cycle of D. minutus in Chapada dos Guimarães is discontinuous and seems related to variations in air temperature and rainfall.

The choice of bromeliads as a microhabitat by Scinax argyreornatus (Anura, Hylidae) / A escolha de bromélias, como micro-habitat, por Scinax argyreornatus (Anura, Hylidae)

The association of anurans to bromeliads presents different degrees of interaction such as: eventual, obligatory and bromeligen. The frog species Scinax argyreornatus shows a regular association with these plants. The goal of this study is to characterise the degree of association between the frog S. argyreornatus to different species of bromeliads. We identified which species of bromeliad is regularly associated with S. argyreornatus and recognised which factors interfere with this association preference. We analysed the Concentration of Relative Dominance of frogs per bromeliad species. As possible criteria for frog association preference to different bromeliads species we established the analysis of leaves number, length and width, number of leaf axils, stored water in the axils, pH and relative humidity, among other organographic components. Our observations were analyzed by ANOVA followed by the Tukey test. We also evaluated the preference for association by the constancy of Bodenheimer. The correlation matrix indicated that the relative humidity is the factor responsible for the frog-bromeliads association, except for Aechmea sp.. However Aechmea sp. was the species with greater constancy of occupation followed by Quesnelia arvensis and Neoregelia johannis. According to our statistical results, Aechmea sp. and Q. arvensis are not different regarding organographic parameters, but differ from N. johannis. Our observations suggest that the bromeliads organographic structure and the relative humidity are key conditions which influence the preferences of S. argyreornatus to bromeliad species, while the other features showed no correlation.

A associação dos anuros às bromeliáceas pode apresentar diferentes graus de interação, como: bromelículas eventuais, bromelículas obrigatórios e bromelígenas. Scinax argyreornatus apresenta uma associação frequente a essas plantas. Propõe-se, neste estudo, qualificar esse grau de associação, buscando identificar qual espécie de bromélia é mais ocupada, reconhecendo os fatores que interferem nessa escolha de ocupação. Analisou-se a Concentração de Dominância Relativa dos anuros por espécie de bromélia, sendo considerados como prováveis critérios de escolha: número, largura e comprimento das folhas, número de axilas foliares, volume de água acumulado, pH e umidade relativa, entre outros componentes organográficos, que foram submetidos à ANOVA e, sempre que observadas diferenças significativas, ao teste de Tukey. Avaliou-se também a escolha de ocupação pela constância de Bodenheimer. A matriz de correlação apontou a UR% como o fator responsável pela ocorrência do animal nas bromélias, exceto para Aechmea sp.; entretanto, esta foi a planta com maior Constância de ocupação, seguida por Quesnelia arvensis e Neoregelia johannis. Segundo a ANOVA, Aechmea sp. e Q. arvensis não apresentam diferenças entre si do ponto de vista organográfico, mas diferem de N. johannis. Foi possível inferir que a estrutura organográfica da planta e a UR% são as condições que interferem na escolha de ocupação das bromélias por S. argyreornatus, enquanto as outras características não demonstraram nenhuma correlação.