RESUMO

Photoreceptor oil droplets (ODs) are spherical organelles placed most commonly within the inner segment of the cone photoreceptors. Comprising neutral lipids, ODs can be either non-pigmented or pigmented and have been considered optically functional in various studies. Among living amphibians, ODs were only reported to occur in frogs and toads (Anura), while they are absent in salamanders and caecilians. Nonetheless, the limited understanding of their taxonomic distribution in anurans impedes a comprehensive assessment of their evolution and relationship with visual ecology. We studied the retinae of 134 anuran species, extending the knowledge of the distribution of ODs to 46 of the 58 currently recognized families, and providing a new perspective on this group that complements the available information from other vertebrates. The occurrence of ODs in anurans shows a strong phylogenetic signal, and our findings revealed that ODs evolved at least six times during the evolutionary history of the group, independently from other vertebrates. Although no evident correlation was found between OD occurrence, adult habits and diel activity, it is inferred that each independent origin involves distinct scenarios in the evolution of ODs concerning photic habits. Furthermore, our results revealed significant differences in the size of the ODs between nocturnal and arrhythmic anurans relative to the length of the cones' outer segment.

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Colour signals play pivotal roles in different communication systems, and the evolution of these characters has been associated with behavioural ecology, integumentary production processes and perceptual mechanisms of the species involved. Here, we present the first insight into the molecular and histological basis of skin colour polymorphism within a miniaturized species of pumpkin toadlet, potentially representing the lowest size threshold for colour polytypism in tetrapods. Brachycephalus actaeus exhibits a coloration ranging from cryptic green to conspicuous orange skin, and our findings suggest that colour morphs differ in their capability to be detected by potential predators. We also found that the distribution and abundance of chromatophores are variable in the different colour morphs. The expression pattern of coloration related genes was predominantly associated with melanin synthesis (including dct, edn1, mlana, oca2, pmel, slc24a5, tyrp1 and wnt9a). Up-regulation of melanin genes in grey, green and brown skin was associated with higher melanophore abundance than in orange skin, where xanthophores predominate. Our findings provide a significant foundation for comparing and understanding the diverse pathways that contribute to the evolution of pigment production in the skin of amphibians.

RESUMO

The present paper comprises a systematic survey of helminths (trematodes, an acanthocephalan and nematodes) found in nine species of freshwater fishes in Ecuador collected in March 1999 and those (a trematode and acanthocephalans) collected from an amphibian and two species of freshwater fishes in Venezuela in 1992, 1996 and 2001. The following 17 helminth species were recorded: Trematoda: Prosthenhystera ornamentosa sp. n., P. obesa (Diesing, 1850), Crassicutis intermedius (Szidat, 1954), C. cichlasomae Manter, 1936 and Glypthelmins eleutherodactyli sp. n. Acanthocephala: Quadrigyrus torquatus Van Cleave, 1920, Gracilisentis variabilis (Diesing, 1851) and Neoechinorhynchus (Neoechinorhynchus) ecuadoris sp. n. Nematoda: Cosmoxynema vianai Travassos, 1949, Travnema travnema Pereira, 1938, Touzeta ecuadoris Petter, 1987, Sprentascaris hypostomi Petter et Cassone, 1984, Sprentascaris sp., Contracaecum sp. Type 1 larvae, Contracaecum sp. Type 2 larvae, Procamallanus (Procamallanus) peraccuratus Pinto, Noronha et Rolas, 1976 and Procamallanus (Spirocamallanus) sp. juv. Nearly all of these parasites are reported from Ecuador or Venezuela for the first time and many of these findings represent new host records. The new species P. ornamentosa sp. n. was collected from the gall-bladder of an unidentified anostomid (Anostomidae, Characiformes) in Ecuador, G. eleutherodactyli sp. n. from the digestive tract of the frog Eleutherodactylus sp. (Eleutherodactylidae, Anura) in Venezuela and N. (N.) ecuadoris sp. n. from the intestine of Lebiasina sp. (Lebiasinidae, Characiformes) in Ecuador. Most parasites are briefly described and illustrated and problems concerning their morphology, taxonomy, hosts and geographical distribution are discussed.

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Horned frogs, members of the Ceratophryidae family, encompass a group of anurans varying in size and behavior, yet unified by morphological and behavioral traits enabling them to adopt a megalophagous diet (i.e., large prey feeding). Although the group has been the focus of numerous studies, our understanding of its feeding behavior remains limited. In this study, we characterize the feeding mechanism in five species representing the three extant genera of ceratophryid anurans, both in terrestrial and aquatic environments. We also explore the ability of Chacophrys pierottii to adjust feeding behavior based on prey type. Our findings show that all species are capable of wide mouth opening, displaying an asymmetric feeding cycle. While tongue usage is the primary method for capturing prey on land, ceratophryids may use their forelimbs to manipulate prey into their mouths, exhibiting different behavioral patterns. C. pierottii shows modulation of its feeding kinematics and is also capable of some modulation of its feeding in response to prey properties.

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During an ecological study with a near-endangered anuran in Brazil, the Schmidt's Spinythumb frog, Crossodactylus schmidti Gallardo, 1961, we were given a chance to analyze the gastrointestinal tract of a few individuals for parasites. In this paper, we describe a new species of an allocreadiid trematode of the genus Creptotrema Travassos, Artigas & Pereira, 1928, which possesses a unique trait among allocreadiids (i.e., a bivalve shell-like muscular structure at the opening of the ventral sucker); the new species represents the fourth species of allocreadiid trematode parasitizing amphibians. Besides, the new species is distinguished from other congeners by the combination of characters such as the body size, ventral sucker size, cirrus-sac size, and by having small eggs. DNA sequences through the 28S rDNA and COI mtDNA further corroborated the distinction of the new species. Phylogenetic analyses placed the newly generated sequences in a monophyletic clade together with all other sequenced species of Creptotrema. Genetic divergences between the new species and other Creptotrema spp. varied from 2.0 to 4.2% for 28S rDNA, and 15.1 to 16.8% for COI mtDNA, providing robust validation for the recognition of the new species. Even though allocreadiids are mainly parasites of freshwater fishes, our results confirm anurans as hosts of trematodes of this family. Additionally, we propose the reallocation of Auriculostoma ocloya Liquin, Gilardoni, Cremonte, Saravia, Cristóbal & Davies, 2022 to the genus Creptotrema. This study increases the known diversity of allocreadiids and contributes to our understanding of their evolutionary relationships, host-parasite relationships, and biogeographic history.

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Changes in environmental temperature may induce variations in thermal tolerance and sensitivity in ectotherm organisms. These variations generate plastic responses that can be analyzed by examining their Thermal Performance Curves (TPCs). Additionally, some performance traits, like locomotion, could be affected by other factors such as biological interactions (e.g., predator-prey interaction). Here, we evaluate if the risk of predation modifies TPCs in Mendoza four-eyed frog (Pleurodema nebulosum, Burmeister, 1861) and Guayapa's four-eyed frog (Pleurodema guayapae, Barrio, 1964), two amphibian species that occur in ephemeral ponds in arid environments. We measured thermal tolerances and maximum swimming velocity at six different temperatures in tadpoles under three situations: control, exposure to predator chemical cues, and exposure to conspecific alarm cues. TPCs were fitted using General Additive Mixed Models. We found that curves of tadpoles at risk of predation differed from those of control mainly in thermal sensitivity parameters. Our work confirms the importance of biotic interactions have in thermal physiology.

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RESUMO

Amphibians are considered bioindicators of the environment due to their high sensitivity and involvement in terrestrial and aquatic ecosystems. In the last two decades, 2,4-D has been one of the most widely used herbicides in Brazil and around the world, as its use has been authorized for genetically modified crops and therefore has been detected in surface and groundwater. Against this background, the aim of this work was to investigate the effects of environmentally relevant concentrations of 2,4-D-based herbicides on survival, malformations, swimming activity, presence of micronuclei and erythrocyte nuclear abnormalities in Physalaemus cuvieri tadpoles. The amphibians were exposed to six concentrations of 2,4-D-based herbicides: 0.0, 4.0, 30.0, 52.5, 75.0, and 100 µg L-1, for 168 h. At concentrations higher than 52.5 µg L-1, significantly increased mortality was observed from 24 h after exposure. At the highest concentration (100 µg L-1), the occurrence of mouth and intestinal malformations was also observed. The occurrence of erythrocyte nuclear abnormalities at concentrations of 30.0, 52.5, 75.0 and 100 µg L-1 and the presence of micronuclei at concentrations of 52.5, 75.0, and 100 µg L-1 were also recorded. These effects of 2,4-D in P. cuvieri indicate that the ecological risk observed at concentrations above 10.35 µg L-1 2,4-D may represent a threat to the health and survival of this species, i.e., exposure to 2,4-D at concentrations already detected in surface waters in the species' range is toxic to P. cuvieri.

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Abstract In order to better understand the ossification processes in anurans our study was carried out on tadpoles and adults of Lithobates catesbeianus. In this sense, we characterized the kinetic properties of alkaline phosphatase with p-nitrophenylphosphatase (pNPP) and pyrophosphate (PPi) and evaluated the activities of tartrate-resistant acid phosphatase and acid phosphatase. The enzyme extracts were obtained from tadpoles and adult femurs, which were divided into epiphysis and diaphysis. After homogenization, the samples were submitted to differential centrifugation to obtain cell membranes and, further, to phospholipase C (PIPLC) treatment, to remove membrane-bound proteins anchored by phosphatidylinositol. The average of specific activity for pNPP hydrolysis (at pH 10.5) by alkaline phosphatase released by phosphatidylinositol-specific phospholipase C (PIPLC) from Bacillus cereus among different bone regions at different animal ages was 1,142.57 U.mg-1, while for PPi hydrolysis (at pH 8.0), it was 1,433.82 U.mg-1. Among the compounds tested for enzymatic activity, the one that influenced the most was EDTA, with approximately 67% of inhibition for pNPPase activity and 77% for PPase activity. In the case of kinetic parameters, the enzyme showed a "Michaelian" behavior for pNPP and PPi hydrolysis. The Km value was around 0.6mM for pNPPase activity and ranged from 0.01 to 0.11mM for PPase activity, indicating that the enzyme has a higher affinity for this substrate. The study of pNPP and PPi hydrolysis by the enzyme revealed that the optimum pH of actuation for pNPP was 10.5, while for PPi, which is considered the true substrate of alkaline phosphatase, was 8.0, close to the physiological value. The results show that regardless of the ossification type that occurs, the same enzyme or isoenzymes act on the different bone regions and different life stages of anurans. The similarity of the results of studies with other vertebrates shows that anurans can be considered excellent animal models for the study of biological calcification.

Resumo Para melhor compreender o processo de ossificação em anuros, nosso estudo foi conduzido em girinos e adultos de Lithobates catesbeianus. Nesse sentido, as propriedades cinéticas da fosfatase alcalina com p-nitrofenilfosfato (pNPP) e pirofosfato (PPi) foram caracterizadas, e as atividades enzimáticas das fosfatases ácida e ácida tartarato resistente foram avaliadas. Os extratos enzimáticos foram obtidos de fêmur de girinos e adultos, divididos em epífise e diáfise. Após a homogeneização as amostras foram submetidas à centrifugação diferencial para obter membrana celular e, em seguida, ao tratamento com fosfolipase C (PIPLC), para remover as proteínas de membrana ancoradas por fosfatidilinositol. A média da atividade específica da fosfatase alcalina, liberada pela PIPLC de Bacillus cereus, para a hidrólise de pNPP (pH 10,5) nas diferentes regiões do fêmur e idades dos animais foi de 1.142,57 U.mg-1, enquanto para a hidrólise do PPi (pH 8,0) foi de 1.433,82 U.mg-1. Entre os compostos testados para a atividade enzimática, o de maior influência foi o EDTA, inibindo aproximadamente 67% e 77% das atividades de pNPPase e PPase, respectivamente. Quanto aos parâmetros cinéticos, a enzima apresentou comportamento Michaeliano para a hidrólise dos dois substratos. O valor de Km foi de 0,6 mM para a atividade de pNPPase e variou de 0,01 a 0,11 para a atividade de PPase, indicando uma maior afinidade por esse substrato. O estudo da hidrólise de pNPP e PPi revelou que o pH ótimo aparente de atuação foi de 10,5 para o pNPP e 8,0 para o PPi, próximo ao fisiológico, sendo que esse é considerado o substrato natural da fosfatase alcalina. Os resultados demonstram que, apesar do tipo de ossificação que ocorre, a mesma enzima ou isoenzimas, atuam nos diferentes locais do osso e estágios de vida dos anuros. A similaridade dos estudos com os realizados com outros vertebrados apontam que os anuros podem ser considerados excelentes modelos animais para o estudo da calcificação biológica.

Assuntos

RESUMO

Abstract In order to better understand the ossification processes in anurans our study was carried out on tadpoles and adults of Lithobates catesbeianus. In this sense, we characterized the kinetic properties of alkaline phosphatase with p-nitrophenylphosphatase (pNPP) and pyrophosphate (PPi) and evaluated the activities of tartrate-resistant acid phosphatase and acid phosphatase. The enzyme extracts were obtained from tadpoles and adult femurs, which were divided into epiphysis and diaphysis. After homogenization, the samples were submitted to differential centrifugation to obtain cell membranes and, further, to phospholipase C (PIPLC) treatment, to remove membrane-bound proteins anchored by phosphatidylinositol. The average of specific activity for pNPP hydrolysis (at pH 10.5) by alkaline phosphatase released by phosphatidylinositol-specific phospholipase C (PIPLC) from Bacillus cereus among different bone regions at different animal ages was 1,142.57 U.mg-1, while for PPi hydrolysis (at pH 8.0), it was 1,433.82 U.mg-1. Among the compounds tested for enzymatic activity, the one that influenced the most was EDTA, with approximately 67% of inhibition for pNPPase activity and 77% for PPase activity. In the case of kinetic parameters, the enzyme showed a Michaelian behavior for pNPP and PPi hydrolysis. The Km value was around 0.6mM for pNPPase activity and ranged from 0.01 to 0.11mM for PPase activity, indicating that the enzyme has a higher affinity for this substrate. The study of pNPP and PPi hydrolysis by the enzyme revealed that the optimum pH of actuation for pNPP was 10.5, while for PPi, which is considered the true substrate of alkaline phosphatase, was 8.0, close to the physiological value. The results show that regardless of the ossification type that occurs, the same enzyme or isoenzymes act on the different bone regions and different life stages of anurans. The similarity of the results of studies with other vertebrates shows that anurans can be considered excellent animal models for the study of biological calcification.

Resumo Para melhor compreender o processo de ossificação em anuros, nosso estudo foi conduzido em girinos e adultos de Lithobates catesbeianus. Nesse sentido, as propriedades cinéticas da fosfatase alcalina com p-nitrofenilfosfato (pNPP) e pirofosfato (PPi) foram caracterizadas, e as atividades enzimáticas das fosfatases ácida e ácida tartarato resistente foram avaliadas. Os extratos enzimáticos foram obtidos de fêmur de girinos e adultos, divididos em epífise e diáfise. Após a homogeneização as amostras foram submetidas à centrifugação diferencial para obter membrana celular e, em seguida, ao tratamento com fosfolipase C (PIPLC), para remover as proteínas de membrana ancoradas por fosfatidilinositol. A média da atividade específica da fosfatase alcalina, liberada pela PIPLC de Bacillus cereus, para a hidrólise de pNPP (pH 10,5) nas diferentes regiões do fêmur e idades dos animais foi de 1.142,57 U.mg-1, enquanto para a hidrólise do PPi (pH 8,0) foi de 1.433,82 U.mg-1. Entre os compostos testados para a atividade enzimática, o de maior influência foi o EDTA, inibindo aproximadamente 67% e 77% das atividades de pNPPase e PPase, respectivamente. Quanto aos parâmetros cinéticos, a enzima apresentou comportamento Michaeliano para a hidrólise dos dois substratos. O valor de Km foi de 0,6 mM para a atividade de pNPPase e variou de 0,01 a 0,11 para a atividade de PPase, indicando uma maior afinidade por esse substrato. O estudo da hidrólise de pNPP e PPi revelou que o pH ótimo aparente de atuação foi de 10,5 para o pNPP e 8,0 para o PPi, próximo ao fisiológico, sendo que esse é considerado o substrato natural da fosfatase alcalina. Os resultados demonstram que, apesar do tipo de ossificação que ocorre, a mesma enzima ou isoenzimas, atuam nos diferentes locais do osso e estágios de vida dos anuros. A similaridade dos estudos com os realizados com outros vertebrados apontam que os anuros podem ser considerados excelentes modelos animais para o estudo da calcificação biológica.

RESUMO

Abstract Specimens (50) of Leptophis nigromarginatus from northern Brazil, Peru, Ecuador and Colombia were examined for the composition of stomach contents. Most prey items were tree frogs, especially those of the family Hylidae (96%). Most of the anurans identified belong to the Scinax ruber species group (14%) and Scinax sp. (8%). Both sexes preyed on small-sized items (1.8-4.4% of snake snout-vent length), but females capture larger prey than males. Leptophis nigromarginatus seems to manipulate captured prey before ingestion since most of the prey items (68%) were swallowed head-first. There is a positive correlation between female snout-vent length and prey size and between and head length and prey size.

Resumo Espécimes (50) de Leptophis nigromarginatus do Norte do Brasil, Peru, Equador e Colômbia foram examinados quanto à composição do conteúdo estomacal. A maioria das presas eram pererecas, especialmente as da família Hylidae (96%). A maioria dos anuros identificados pertence ao grupo de espécies Scinax ruber (14%) e Scinax sp. (8%). Ambos os sexos predaram itens de pequeno porte (1,8-4,4% do comprimento focinho-cloaca da serpente), mas as fêmeas capturam presas maiores que os machos. Leptophis nigromarginatus parece manipular as presas capturadas antes da ingestão, uma vez que a maioria das presas (68%) foi engolida pela cabeça. Há uma correlação positiva entre o comprimento do focinho-cloaca da fêmea e o tamanho da presa e entre o comprimento da cabeça e o tamanho da presa.