Metabolic changes in the subtropical frog Boana pulchella during experimental cooling and recovery conditions.

Frogs have developed biochemical and physiological adaptations to occupy diverse ecological niches on Earth successfully. Survival in frozen states is a fascinating strategy made possible by evolving adaptations to produce cryoprotectant solutes. The hylid frog Boana pulchella thrives in South American regions with cold climates, remaining active while enduring sporadic subzero temperatures during winter. The species' metabolic changes during subzero exposure remain unclear. Therefore, we exposed B. pulchella to cooling and recovery, assessing plasma and tissue metabolite changes. Cooling significantly reduced urea concentrations in plasma (P = 0.033), muscle (P = 0.001), heart (P = 0.009), and brain (P = 0.041) compared to acclimation. Liver glucose oxidation and glycogen synthesis were lower in cooling and recovery than in acclimation (P < 0.0001 and P = 0.0117, respectively). Muscle glycogen synthesis was lower in recovery than acclimation (P = 0.0249). These results demonstrate B. pulchella's physiological strategies during subzero exposure, likely reflecting species-specific evolutionary adaptations for brief subzero exposures that enable winter survival in its natural habitat.

Hematologic variables of free-living Leptodactylus luctator with and without hemoparasites and thrombidiform mites in southern Brazil.

It has been suggested that anuran amphibian parasites can cause clinical signs in situations of environmental imbalance. In the family Leptodactylidae, information about hematology is scarce, although these are well-known tools for the diagnosis and prognosis in clinical practice and potential bioindicators of environmental stress. The objective of this study is to describe Leptodactylus luctator hematology, to report the occurrence of hemoparasites and thrombidiform mites, and to compare the hematological variables under the presence and absence of these organisms. Ectoparasites and heparinized blood samples from 40 free-living specimens of L. luctator were collected for analysis. Hematologic variables and total plasma protein were compared between groups with and without hemoparasites and intradermal mites. As results, structures compatible with hemogregarines, Lankesterella sp., five morphotypes of Trypanosoma spp., microfilaria, Aegyptianella sp., an unidentified intraleukocytic hemoparasite, and frog erythrocytic virus (FEV) inclusion bodies were identified in the blood samples, besides Hannemania spp. intradermal mites. The hemoparasite occurrence was higher than previously reported in other anuran families and locations. Also, L. luctator has smaller red blood cells (RBCs) and white blood cells (WBCs), and a hyposegmentation of the neutrophil nucleus, when compared to many other amphibians. White blood cell, neutrophil, and monocyte counts were higher in animals parasitized by mites. There was no correlation between the number of parasitized RBCs and hematologic variables. This study provides anuran hematologic information, in addition to indicating a host reaction to infestation by Hannemania spp. mites, besides constituting the first record of the distribution of hemoparasites and intradermal mites in L. luctator of the study region.

Seasonal variations in the intermediate metabolism in South American tree-frog Boana pulchella.

Seasonal metabolic changes can be observed in many anurans' species. In subtropical environments with environmental temperatures variations, the temperature is a factor that can influence the extent and intensity of activity in many anuran species. Nonetheless, some species of subtropical frogs may remain active throughout the year. Boana pulchella, a subtropical species, seems to be able to survive low temperatures and remain reproductively active even in the coldest months. Therefore, we hypothesized that B. pulchella presents seasonal changes in the energy metabolism to sustain activity during all year. This study evaluated the main energy substrate levels and metabolism of B. pulchella in plasma, liver and muscle of male individuals collected in winter, spring, summer and fall in the state of Rio Grande do Sul, Brazil. Our results showed that B. pulchella has a higher glycolytic oxidation rate in liver (P = 0.0152) and muscle (P = 0.0003) and higher glycogenesis from glucose in muscle (P = 0.0002) in summer, indicating the main energy substrates in this season is glucose. The higher muscle glycogen (P = 0.0008) and lower plasma glucose in fall (P = 0.0134) may indicate an anticipatory regulation for storing to the most thermally demanding cold period: winter. These results indicated seasonal differences in the main energy substrates, and these metabolic changes among seasons can be part of a metabolic adjustment allowing maintenance of reproductive activity all year in Boana pulchella species.

Are the unken reflex and the aposematic colouration of Red-Bellied Toads efficient against bird predation?

Aposematic signals as well as body behaviours may be important anti-predator defences. Species of the genus Melanophryniscus are characterised by having toxic lipophilic alkaloids in the skin and for presenting a red ventral colouration, which can be observed when they perform the behaviour called the unken reflex. Both the reflex behaviour and the colouration pattern are described as defence mechanisms. However, there are currently no studies testing their effectiveness against predators. This study aimed to test experimentally if both ventral conspicuous colouration and the unken reflex in Melanophryniscus cambaraensis function as aposematic signals against visually oriented predators (birds). We simulated the species studied using three different clay toad models as follows: (a) in a normal position with green coloured bodies, (b) in the unken reflex position with green coloured body and extremities and (c) in the unken reflex position with a green body and red extremities. Models were distributed on a known M. cambaraensis breeding site and in the adjacent forest. More than half of the attacks on the models were from birds; however, there was no preference for any model type. Thus, just the presence of the red colour associated with the motionless unken reflex position does not seem to prevent attacks from potential predators. It is possible that the effective aposematic signal in Melanophryniscus is achieved through the unken reflex movement together with the subsequent exhibition of the warning colouration and the secretion of toxins.

Anuran road-kills neighboring a peri-urban reserve in the Atlantic Forest, Brazil.

Mortality from road-kills may figure among the important causes of decline in amphibian populations and species extinctions worldwide. Evaluation of the magnitude, composition, and temporal and spatial distributions of amphibian road-kills is a key step for mitigation planning, especially in peri-urban reserves. Once a month for 16 months, we surveyed, on foot, a 4.4 km section of state road ERS-389 bordering the Itapeva reserve in the southern Atlantic Forest. We recorded 1433 anuran road-kills and estimated a mortality rate of 9002 road-kills/km/year. The species most often recorded were the largest ones: Leptodactylus latrans, Rhinella icterica, Leptodactylus gracilis and Hypsiboas faber; 54.5% of the carcasses could not be identified. Anuran mortality was concentrated in summer, and was associated with temperature, rainfall and photoperiod. Leptodactylus road-kills were strongly influenced by vehicle traffic, probably because of its high abundance during the entire study period. Road-kill hotspots differed for anurans as a group and for single species, and we found an association among spatial patterns of mortality and types of land cover, distance from the nearest waterbody, roadside ditches, and artificial light. Traffic should be banned temporarily during periods of high mortality, which can be forecasted based on meteorological data. A comprehensive mitigation approach should take into account hotspots of all anuran records, and also of target species for selecting locations for amphibian passages and fencing. Roadside ditches, artificial waterbodies, and conventional street lights should be reduced as much as possible, since they may represent ecological traps for anuran populations.

Composição e ameaças à conservação dos anfíbios anuros do Parque Estadual de Itapeva, município de Torres, Rio Grande do Sul, Brasil / Composition and threats for conservation of anuran amphibians from Itapeva State Park, Municipality of Torres, Rio Grande do Sul, Brazil

A obtenção de listas de espécies constitui-se no primeiro passo para o desenvolvimento de estratégias de conservação. O Parque Estadual de Itapeva (PEVA) é uma unidade de conservação localizada numa estreita faixa entre a RS-389 (Estrada do Mar), e a praia de Itapeva, no município de Torres, extremo norte da planície costeira do Rio Grande do Sul (29º 21' S e 49º 45' W). No Parque, observa-se a presença de grandes dunas móveis, dunas fixas, floresta paludosa, campos, floresta de restinga, banhados e riachos. Os objetivos deste trabalho são: fornecer uma listagem de espécies de anuros do PEVA, informar os ambientes em que foram encontradas e identificar as ameaças a estes anfíbios na área do Parque. O inventariamento da anurofauna estendeu-se de março de 2000 a março de 2003. As campanhas tiveram freqüência mensal no período de maio de 2000 a março de 2001, e após este período foram realizadas oito campanhas adicionais. As amostragens tiveram a duração média de duas noites e dois dias, envolvendo a visita periódica a todos os ambientes com ocorrência potencial de anfíbios anuros, concentrando-se em áreas alagadas, açudes, poças temporárias, arroios, córregos e outros corpos d'água. As coletas noturnas geralmente iniciavam-se às 20 horas e terminavam às 23 horas. Durante o dia os anuros eram procurados sob troncos, pedras, telhas e em locais próximos a residências. Oito ambientes foram amostrados: dunas primárias (DP), banhados (BD), dunas móveis (DM), dunas fixas (DF), campos alagadiços (CA), capoeira (CP), mata paludosa (MP) e campos secos (CS). Foram registradas 29 espécies de anuros das famílias Hylidae (11 spp.), Leiuperidae (6 spp.), Bufonidae (4 spp.), Leptodactylidae (3 spp.), Brachycephalidae (2 spp.), Cycloramphidae (1 sp.), Microhylidae (1 sp.) e Ranidae (1 sp.). O PEVA, até o momento, é o local com o terceiro maior número de espécies de anuros do Rio Grande do Sul. Duas espécies estão ameaçadas de extinção no Rio Grande do Sul...

A species inventory is the first step to develop conservation strategies. The Itapeva State Park (PEVA) is a protected area located in a narrow zone between RS-389 (Estrada do Mar) and the Itapeva beach, near Torres city, extreme north of the coastal plain of the Rio Grande do Sul State (29º 21' S and 49º 45' W). The Park is covered by mobile dunes, fixed dunes, marshy forest, grasslands, dry forests, wetlands and streams. The aim of this study is to list the anuran species of PEVA, to describe their habitats, and to identify possible impact on these anurans inside the Park. The anuran fauna survey occurred from March 2000 to March 2003. The field trips were monthly from May de 2000 to March 2001 after this period, eigth adicional field trips were done. The surveys comprised two nights and two days. The sampling methods included periodic visits to all environments with potential anuran incidence, wet areas, pools, temporary pools, streams and wetlands. During the day we looked for animals under logs, stones, tiles, and near houses. Nocturnal collections were from 8:00 to 11:00 PM. Eight environments were sampled: primary sand dunes (DP), wetlands (BD), movel dunes (DM), fixed dunes (DF), wet grasslands (CA), "capoeira" (regenerating forest), (CP), marshy forest (MP) and dry grasslands (CS). Twenty-nine anuran species from eight families were recorded: Hylidae (11 spp.), Leiuperidae (6 spp.), Bufonidae (4 spp.), Leptodactylidae (3 spp.), Brachycephalidae (2 spp.), Cycloramphidae (1 sp.), Microhylidae (1 sp.) e Ranidae (1 sp.). To date the PEVA is the area with the third highest number of species in Rio Grande do Sul State.Two species are threatened in Rio Grande do Sul: Haddadus binotatus and Melanophryniscus dorsalis. The main observed threats were: forest drainage, forest fragmentation, presence of invasive species and wetland degradation.

Assessment of the freshwater annual fish Cynopoecilus melanotaenia as a toxicity test organism using three reference substances.

This study presents a preliminary evaluation of the use of the Brazilian fish Cynopoecilus melanotaenia as a test organism in toxicity tests. The cryptobiotic stage presented by the eggs of fish C. melanotaenia can overcome the difficulty of continuously keeping cultures and recruiting healthy animals in sufficient numbers to be used in toxicity tests. In order to determine the applicability of this species as a test organism, three different reference substances were evaluated in 96-h acute toxicity tests: Copper sulfate (CuSO4 x 5H2O), sodium dodecil sulfate (C12H25NaO4S), and sodium chloride (NaCl). Sensitivity ranged as follows: copper sulfate (0.05-0.13 mg/L), sodium dodecil sulfate (10.7-19.0 mg/L), and sodium chloride (1.44-1.96 g/L). We conclude that C. melanotaenia shows potential as a test organism in toxicity tests; however, further research should be conducted with other substances and should be compared with the research on other species before we can reach more conclusive results.