A Phylogenetic Lineage of Closely Related Trypanosomes (Trypanosomatidae, Kinetoplastida) of Anurans and Sand Flies (Psychodidae, Diptera) Sharing the Same Ecotopes in Brazilian Amazonia

Analysis of the phylogenetic relationships among trypanosomes from vertebrates and invertebrates disclosed a new lineage of trypanosomes circulating among anurans and sand flies that share the same ecotopes in Brazilian Amazonia. This assemblage of closely related trypanosomes was determined by comparing whole SSU rDNA sequences of anuran trypanosomes from the Brazilian biomes of Amazonia, the Pantanal, and the Atlantic Forest and from Europe, North America, and Africa, and from trypanosomes of sand flies from Amazonia. Phylogenetic trees based on maximum likelihood and parsimony corroborated the positioning of all new anuran trypanosomes in the aquatic clade but did not support the monophyly of anuran trypanosomes. However, all analyses always supported four major clades (An01-04) of anuran trypanosomes. Clade An04 is composed of trypanosomes from exotic anurans. Isolates in clades An01 and An02 were from Brazilian frogs and toads captured in the three biomes studied, Amazonia, the Pantanal and the Atlantic Forest. Clade An01 contains mostly isolates from Hylidae whereas clade An02 comprises mostly isolates from Bufonidae; and clade An03 contains trypanosomes from sand flies and anurans of Bufonidae, Leptodactylidae, and Leiuperidae exclusively from Amazonia. To our knowledge, this is the first study describing morphological and growth features, and molecular phylogenetic affiliation of trypanosomes from anurans and phlebotomines, incriminating these flies as invertebrate hosts and probably also as important vectors of Amazonian terrestrial anuran trypanosomes.

Leishmanicidal Activity and Immobilization of dermaseptin 01 antimicrobial peptides in ultrathin films for nanomedicine applications

Antimicrobial peptides (AMPs) are essential for the innate immune system of eukaryotes, imparting protection against pathogens and their proliferation in host organisms. The recent interest in AMPs as active materials in bionanostructures is due to the properties shown by these biological molecules, such as the presence of an alpha-helix structure and distribution of positive charges along the chain. In this study the antimicrobial peptide dermaseptin 01 (DS 01), from the skin secretion of Phyllomedusa hypochondrialis frogs was immobilized in nanostructured layered films in conjunction with nickel tetrasulfonated phthalocyanines. The leishmanicidal activity of DS 01 was confirmed using kinetic essays, in which DS 01 promoted death of all metacyclic promastigote cells in 45 minutes. Surprisingly, the immobilized DS 01 molecules displayed electroactivity, as revealed by electrochemical experiments, in which an oxidation peak at about 0.61 V was observed for a DS 01 monolayer deposited on top of a conductive electrode. Such electroactivity was used to investigate the sensing abilities of the nanostructured films toward Leishmania. We observed an increase in the oxidation current as a function of number of Leishmania cells in the electrolytic solution at concentrations down to 10(3) cells/mL. The latter is indicative that the use of AMPs immobilized in electroactive nanostructured films may be of interest for applications in the pharmaceutical industry and diagnosis.

Desempenho zootécnico de girinos de rã-touro com diferentes níveis de proteína e energia digestíveis

A partir dos valores de digestibilidade de alguns ingredientes foram propostas dietas, com diferentes níveis de proteína e energia digestíveis, em esquema fatorial 3 x 3, destinadas a girinos de rã-touro para avaliar o desempenho zootécnico. O experimento, com duração de 60 dias, foi conduzido com 810 girinos, oriundos da mesma desova, distribuídos em 27 caixas de polipropileno, em uma densidade de 1girino/L. O delineamento experimental foi um esquema fatorial com três níveis de proteína digestível (27; 31 e 35%) e três níveis de energia digestível (2.700; 2.800 e 2.900kcal/kg) com três réplicas. Os parâmetros avaliados aos 60 dias foram ganho de peso, consumo da dieta, consumo em proteína da dieta, conversão alimentar, taxa de eficiência proteica e índice de sobrevivência. Os resultados foram submetidos à análise de variância e ao teste de Duncan (95% de precisão). Os resultados encontrados não apresentaram interação entre proteína e energia digestível para nenhum parâmetro avaliado. em relação à proteína digestível, foram verificadas diferenças nos parâmetros consumo em proteína da dieta e taxa de eficiência proteica (TEP), o que demonstra ineficiência das dietas com altos valores proteicos. Deve-se oferecer aos girinos de rã-touro dietas com 27% de proteína digestível, por apresentarem melhor resultado para taxa de eficiência proteica e para consumo em proteína da dieta.

Ecologia trófica de anfíbios anuros: relações filogenéticas em diferentes escalas

Understand the origin, maintenance and the mechanisms that operate in the current biodiversity is the major goal of ecology. Species ecology can be influenced by different factors at different scales. There are three approaches about the ecological differences between species: the first brings that differences result from current processes on niche characteristics (e.g. diet, time, space); the second that species differences are explained by random patterns of speciation, extinction and dispersion, the third that historical events explain the formation and composition of species in communities. This study aims to evaluate the influence of phylogenetic relationships in determining ecological characteristics in amphibians (globally) and test with that, if ecological differences between species of frogs are the result of ancient pre-existing differences or as result of current interactions. Another objective of this study is to verify if ecological, historical or current characteristics determine the size of species geographical distribution. The diet data for analysis of trophic ecology were collected from published literature. We performed a non-parametric MANOVA to test the existence of phylogenetic effects in diet shifts on frogs history. Thus, it is expected to know the main factors that allow the coexistence of anuran species. We performed a phylogenetic regression to analyze if niche breadth, body size and evolutionary age variables determine the size of the geographical distribution of amphibians in the Amazon. In the present study, new contributions to knowledge of major ecological patterns of anurans are discussed under a phylogenetic perspective

Interaction between breeding habitat and elevation affects prevalence but not infection intensity of Batrachochytrium dendrobatidis in Brazilian anuran assemblages

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Haematologic and immunologic parameters of bullfrogs, Lithobates catesbeianus, fed probiotics

The effects of two probiotics (P(1) - Lactobacillus acidophilus, Bifidobacterium bifidum and Enterococcus faecium and P(2) - Bacillus subtilis) supplemented to commercial feed (40% crude protein) on the haematological and immunological parameters of the bullfrog Lithobates catesbeianus were studied. Two doses of each probiotic (5 and 10 g kg-1 of food) were added to the diets and fed to frogs, totalling five treatments over 112 days. Haematological analyses consisted of total and differential leucocyte counts, erythrocyte and thrombocyte counts, haematocrit, haemoglobin levels and RBC indices (mean corpuscular volume, mean corpuscular haemoglobin - and mean corpuscular haemoglobin concentration) and the immunological parameters included phagocytic capacity and phagocytic index of peritoneal phagocytes. The results showed that the probiotics did not significantly influence any of the haematological parameters measured. However, immunological assays showed that the probiotics had an immunostimulating effect. The greatest effects were seen with probiotic P(1) fed at a dose of 10 g kg-1 of diet and probiotic P(2) fed at 5 g kg-1 of diet.

Influência da incorporação de vitamina C à dieta no desempenho produtivo de rãs-touro Lithobates catesbeianus pós-metamorfoseadas

The experiment was developed in Marta Farm, Tremembe city, located in the State of São Paulo - Brazil, using a frogculture commercial project. The aim of this study was evaluate the performance (growth, survival and feed conversion) of bullfrogs Lithobates catesbeianus, post-metamorphosed fed with diets containing different amounts of vitamin C and your performance with the temperature. The animals were kept in 24 boxes of (2.0 x 1.0 m) with an initial density of 50 animals per square meter. We tested the following levels of supplementation in the diet: 0, 250, 500, 750, 1,000 and 2,000 mg vitamin C kg(-1) of diet. The experimental design was completely randomized with six treatments and four replicates. The parameters evaluated were: feed intake, weight gain, feed conversion and survival tax. The animals were weighed at the beginning of the experiment and every 30 days and recorded daily to room temperature. The percentage of food intake decreased with the growth of the frogs, from 3.85 to 1.2% of the live weight, and the mean of temperature decreased from 28 to 20.1 degrees C. It was concluded that there was no influence of vitamin C on the performance (growth, survival and feed conversion) of bullfrogs in the growing stage.

Atividades comportamentais e comportamento alimentar de rã touro, Lithobates catesbeianus (Shaw, 1802), de pigmentação normal e albina em cativeiro

The behavior of bullfrogs reared in captivity must be well understood to support management practices that use efficient feeding regimes. In general, bullfrogs reared in captivity have normal pigmentation, but to develop an enhanced product, some studies have investigated the introduction of albino individuals in frog farms. The present study characterized the behavior of both pigmented and albino bullfrogs reared in captivity. In an initial experiment, 48 bullfrogs (70.5 ± 25.6 g) housed in small stalls were fed once a day at random times. Frogs were filmed and the images showed that both the pigmented and albino varieties behaved similarly: food intake was more frequent at dawn followed by light periods; moving and resting in dry areas may be associated to feeding events; frogs appeared to anticipate feeding time and to rest in the water more frequently in periods other than feeding time; daylight is the recommended period for feeding both pigmented and albino frogs. In a second experiment, 72 albino bullfrogs were fed at fixed times (10 a.m. or 4 p.m.) in small stalls. An initial weight of 23.8 ± 7.6 g was considered to evaluate frog performance, and after the animals reached 60.0 ± 20.0 g, they were filmed for behavior analyses. Food intake varied as a function of feeding time, and frogs were more likely to eat during the early hours of the day and immediately after receiving fresh food. Frogs fed only in the afternoon changed their behavior. Food supplied twice a day stimulated the albino frogs to eat at different times, but did not increase growth. Although fresh food stimulated feeding behavior, food intake was more frequent at dawn. Food supplied at this time of day should therefore be further investigated. The results did not indicate a more suitable feeding time (10 a.m. or 4 p.m.) for albino bullfrogs, or any advantage in using two feedings per day. The results provide xvi important information about bullfrogs in terms of food supply regime and activity preferences throughout the day. This novel information will contribute to future studies in this area

Relação de variáveis ambientais em baias cobertas com polietileno e desempenho da rã-touro (Rana catesbeiana)

Este trabalho visou a relacionar variáveis ambientais em instalação para criação de rãs, com cobertura de polietileno e baias construídas usando material alternativo, com o desempenho de rãs-touro (Rana catesbeiana). No interior das baias, foram medidas as temperaturas do piso, do ar ambiente (bulbo seco), de bulbo úmido, globo negro e da água do reservatório. Foram utilizados 60 animais por baia e três baias por galpão. As variáveis de desempenho estudadas foram peso vivo, ganho de peso e conversão alimentar. Nas condições experimentais, quando a temperatura do ar atingiu valores abaixo de 10 ºC ou superiores a 40 ºC, houve diminuição no consumo de ração pelos animais. Concluiu-se que o estresse predominante, neste tipo de estrutura, para as condições climáticas do período experimental, foi devido, principalmente, às baixas temperaturas. Concluiu-se, ainda, que o uso do Índice de Temperatura e Umidade (THI), na estimativa de variáveis de desempenho, melhorou a precisão da estimativa em relação ao uso exclusivo da temperatura do ar, embora valores desse índice, considerados estressantes para animais superiores, não o tenham sido para as rãs.

Efeito da temperatura sobre o desempenho da rã-touro (Rana catesbeiana Shaw, 1802)

Este estudo foi realizado para avaliar os efeitos do ambiente sobre a performance de rã-touro (Rana catesbeiana Shaw, 1802), criada em gaiolas de fibra de vidro instaladas no interior de estufas climatizadas. Após um período inicial de 15 dias de adaptação às instalações, à temperatura constante de 25,0ºC, os seguintes tratamentos foram aplicados: temperaturas de 23,0; 26,0; 29,0; 32,0; e 35,0ºC, por 30 dias, para rãs com 100 g PV inicial; 24,5; 26,0; 27,5; 29,0; 30,5; e 32,0ºC, também por 30 dias, para rãs com 20 g PV inicial; e a combinação das temperaturas de 26,0 e 29,0ºC com os fotoperíodos de 8, 12 e 16 h de luz a cada 24 horas, para rãs com 100 g PV inicial. Derivando-se as equações de regressão que explicam os efeitos de temperatura sobre o desempenho das rãs, estimaram-se melhores ganhos de peso à temperatura de 27,6e30,1ºC, para rãs com 100 e 20 g PV inicial, respectivamente, com melhor crescimento a 28,2ºC, para as rãs de 100 g PV inicial, e a 29,7ºC, para as rãs de 20 g PV inicial. A temperatura interagiu com fotoperíodo nos seus efeitos sobre ganho de peso e crescimento corporal, peso e rendimento de carcaça, consumo de alimentos e conversão alimentar.

Efeito da temperatura e do fotoperíodo sobre o desenvolvimento do aparelho reprodutor de rã-touro (Rana catesbeiana Shaw, 1802)

Foram avaliados os efeitos da temperatura e do fotoperíodo sobre a maturação sexual de rãs-touro pesando 94,22 g ± 12,03, mantidas durante trinta dias em temperaturas de 20, 23, 26, 29, 32 e 35°C, com fotoperíodo de 12/12 horas de luz/horas de escuridão (h L/E). A temperatura afetou os pesos do corpo gorduroso e do fígado, os quais variaram de acordo com modelos quadráticos, estimando-se maiores pesos de corpo gorduroso a 27,27°C e de fígado a 26,81°C. Estimaram-se ovários mais pesados a 28,36°C e ovidutos mais pesados a 28,77°C. Temperatura afetou a maturação sexual das rãs, avaliada por índices numéricos. Num experimento mais longo, rãs com peso médio inicial de 95,31 ± 8,46 g foram submetidas à combinação das temperaturas de 26 e 29°C com os fotoperíodos de 8/16, 12/12 e 16/8 h L/E, até atingirem a maturidade gonadal. Temperatura interagiu com fotoperíodo em seus efeitos sobre o desenvolvimento dos órgãos reprodutivos de rã-touro. Temperatura afetou a relação diâmetro do abdômen/distância entre os olhos, com maiores valores calculados para 26°C. Verificou-se que os maiores diâmetros dos ovócitos são obtidos a uma temperatura de 26°C, com fotoperíodo de 12,6/11,4 h L/E.

Estufas climatizadas para experimentos ambientais com rãs, em gaiolas

Foram construídas seis estufas climatizadas, instaladas inicialmente no Ranário Experimental da Universidade Federal de Viçosa (UFV) e, posteriormente, no Ranário Experimental da Fundação Universidade Federal do Rio Grande, com o objetivo de realizar experimentos para avaliar os efeitos do ambiente sobre o desempenho de rãs em gaiolas de fibra de vidro. Ambientes com temperaturas de 25ºC e fotoperíodo de 12/12 horas de luz/horas de escuridão (h L/E) serviram para adaptação das rãs por 15 dias antes de cada experimento. Os tratamentos consistiram em simular ambientes com temperaturas variando de 20 a 35ºC e fotoperíodos de 8/16, 12/12 e 16/8 h L/E. Foram realizados experimentos com rã-touro (Rana catesbeiana Shaw, 1802) e rã-manteiga (Leptodactylus ocellatus Linnaeus, 1758). Nessas estufas foi possível estimar que: a) os maiores ganhos de peso de rã-touro foram obtidos entre 27,6 e 29,7ºC, com melhor crescimento entre 28,2 e 30,1ºC; para rã-manteiga os melhores ganhos e conversão alimentar foram observados a 28,6 e 28ºC, respectivamente; b) a temperatura interage com fotoperíodo sobre o desempenho das rãs e seu desenvolvimento gonadal; c) a 27,7ºC (temperatura de conforto térmico) haverá menos rãs dentro d'água; d) a maior temperatura cloacal de rã-touro, 32,1ºC no seco e 33,8ºC dentro d'água, a 35ºC, evidenciou que as rãs se termorregulam; e) os níveis de tetraiodotironina (T4) no plasma decrescem na temperatura de conforto térmico; f) rã-manteiga condiciona-se ao manejo de rotina, reunindo-se ao redor do cocho na hora da alimentação.

Thermal relationships and exercise physiology in anuran amphibians: Integration and evolutionary implications

Thermal and water balance are coupled in anurans, and species with particularly permeable skin avoid overheating more effectively than minimizing variance of body temperature. In turn, temperature affects muscle performance in several ways, so documenting the mean and variance of body temperature of active frogs can help explain variation in behavioral performance. The two types of activities studied in most detail, jumping and calling, differ markedly in duration and intensity, and there are distinct differences in the metabolic profile and fiber type of the supporting muscles. Characteristics of jumping and calling also vary significantly among species, and these differences have a number of implications that we discuss in some detail throughout this paper. One question that emerges from this topic is whether anuran species exhibit activity temperatures that match the temperature range over which they perform best. Although this seems the case, thermal preferences are variable and may not necessarily reflect typical activity temperatures. The performance versus temperature curves and the thermal limits for anuran activity reflect the thermal ecology of species more than their systematic position. Anuran thermal physiology, therefore, seems to be phenotypically plastic and susceptible to adaptive evolution. Although generalizations regarding the mechanistic basis of such adjustments are not yet possible, recent attempts have been made to reveal the mechanistic basis of acclimation and acclimatization. (C) 2007 Elsevier B.V. All rights reserved.

The evolution of jumping performance in anurans: morphological correlates and ecological implications

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)