Avaliação da atividade antimicrobiana de sistemas emulsionados contendo óleos naturais para o tratamento de infecções cutâneas

Natural oils have shown a scientific importance due to its pharmacological activity and renewable character. The copaiba (Copaifera langsdorffii) and Bullfrog (Rana catesbeiana Shaw) oils are used in folk medicine particularly because the anti-inflammatory and antimicrobial activities. Emulsion could be eligible systems to improve the palatability and fragrance, enhance the pharmacological activities and reduce the toxicological effects of these oils. The aim of this work was to investigate the antimicrobial activity of emulsions based on copaiba (resin-oil and essential-oil) and bullfrog oils against fungi and bacteria which cause skin diseases. Firstly, the essential oil was extracted from copaiba oil-resin and the oils were characterized by gas chromatography coupled to a mass spectrometry (GC-MS). Secondly, emulsion systems were produced. A microbiological screening test with all products was performed followed (the minimum inhibitory concentration, the bioautography method and the antibiofilm determination). Staphylococcus aureus, S. epidermidis, Pseudomonas aeruginosa, Candida albicans, C. parapsilosis, C. glabrata, C. krusei and C. tropicalis American Type Culture Collection (ATCC) and clinical samples were used. The emulsions based on copaiba oil-resin and essential oil improved the antimicrobial activity of the pure oils, especially against Staphylococcus e Candida resistant to azoles. The bullfrog oil emulsion and the pure bullfrog oil showed a lower effect on the microorganisms when compared to the copaiba samples. All the emulsions showed a significant antibiofilm activity by inhibiting the cell adhesion. Thus, it may be concluded that emulsions based on copaiba and bullfrog oils are promising candidates to treatment of fungal and bacterial skin infections

Chemical Characterization and Antimicrobial Activity Evaluation of Natural Oil Nanostructured Emulsions

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Apparent digestibility of different ingredients in diets for bullfrog Rana catesbeiana tadpoles

The objective of this study was to determine apparent digestibility of protein and energy of ingredients commonly present in rations fed to bullfrog tadpoles Rana catesbeiana such as corn, soybean meal, wheat meal, rice bran, poultry by-product meal, blood meal, fish meal, corn starch and soybean oil. A completely randomized design was used to test ten experimental diets with four replicates. About 30% of the reference diet was replaced by the test ingredients, except for soybean oil that was used at 10% ratio and 0.5% of chromium oxide as inert marker. The values of apparent digestibility coefficient of crude protein (ADCCP) and gross energy (ADCGE) of the protein ingredients were, respectively: fish meal, 81 and 95%; poultry byproduct meal, 82 and 87%; blood meal, 64 and 53%; and soybean meal, 93 and 90%. While for the energetic ingredients, ADCCP and ADCGE, were, respectively: wheat meal, 100 and 72%; rice bran, 57 and 24%; and corn, 84 and 92%. Soybean oil and cornstarch ADCGE values were 70 and 96%, respectively; cornstarch digestibility is highly dependent on feed processing technique. The results showed that the ing redients of plant origin, except for rice bran, had good digestibility, thus indicating that tadpoles can effectively use plant protein. In this trial, the least digestible ingredients were blood meal and rice bran, thus indicating that the use of such ingredients in bullfrog tadpole diets should be limited. © Copyright by the World Aquaculture Society 2005.

Desenvolvimento biotecnológico de uma emulsão de uso tópico a base de óleo de rã-touro Rana catesbeiana Shaw

The skin is one of the largest organs of the human body and accounts for about 16% of body weight. The body protection against the external environment microorganisms is one of its most important functions, however is necessary that the skin remain intact for this function be exercised, so that when there is an injury on the skin, the process of restructuring needs to be starts, however this restructuration may also be compromised due to some diseases, justifying even more the need for the development of topical products that promote or accelerate the skin healing. Thus the aim of this study was to extract bullfrog oil and to develop a suitable topical emulsion. Two different oil samples were extracted by hot or organic solvent process. Titration techniques and gas chromatography- mass spectrometry were used to characterize the bullfrog oil. The required hydrophile-lipophile balance (HLBr) of bullfrog oil was determined and a pseudo-ternary phase diagram was constructed. The stability of the topical emulsion was evaluated. Then, cellular viability was determined by MTT assay using normal fibroblasts (3T3) and melanoma (B16F10) cells lines. The hot extraction yield was 60.6%. The major polyunsaturated compounds found were Eicosapentaenoic acid (17.6%) and Arachidonic acid (8.4%). HLBr study demonstrated the presence of stable systems with HLB ranging from 12.1 to 13.5 and the pseudo-ternary phase diagram showed mainly emulsion systems (62%). Topical emulsion showed 390 nm, polydispersity 0.05, zeta potential -25 mV and remained stable for ninety days. The bullfrog oil and topical emulsion did not showed citotoxicity in normal fibroblasts cells. However, these systems showed significantly inhibition of melanoma cells growth. In conclusion, the bullfrog oil presented desirable chemical characteristics required to be used for the development of a pharmaceutical and cosmetic products.

Testosterone-immunopositive primordial germ cells in the testis of the bullfrog, Rana catesbeiana

In amphibia, steroidogenesis remains quiescent in distinct seasonal periods, but the mechanism by which spermatogenesis is maintained under low steroidogenic conditions is not clear. In the present study, testosterone location in the testes of Rana catesbeiana was investigated immunohistochemically during breeding (summer) and nonbreeding (winter) periods. In winter, the scarce interstitial tissue exhibited occasional testosterone immunopositivity in the interstitial cells but the cytoplasm of primordial germ cells (PG cells) was clearly immunopositive. By contrast, in summer, PG cells contained little or no immunoreactivity whereas strong immunolabelling was present in the well-developed interstitial tissue. These results suggest that PG cells could retain testosterone during winter. This androgen reservoir could be involved in the control of early spermatogenesis in winter and/or to guarantee spermiogenesis and spermiation in the next spring/summer. The weak or negative immunoreaction in the summer PG cells might reflect consumption of androgen reservoir by the intense spermatogenic activity from spring to summer. Thus, besides acting as stem cells, PG cells of R. catesbeiana could exert an androgen regulatory role during seasonal spermatogenesis.

The oleoresin secretory system in seedlings and adult plants of copaiba (Copaifera langsdorffii Desf., Leguminosae-Caesalpinioideae)

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

Ectopic mineralization of articular cartilage in the bullfrog Rana catesbeiana and its possible involvement in bone closure

Mineralization of the articular cartilage is a pathological condition associated with age and certain joint diseases in humans and other mammals. In this work, we describe a physiological process of articular cartilage mineralization in bullfrogs. Articular cartilage of the proximal and distal ends of the femur and of the proximal end of the tibia-fibula was studied in animals of different ages. Mineralization of the articular cartilage was detected in animals at 1 month post-transformation. This mineralization, which appeared before the hypertrophic cartilage showed any calcium deposition, began at a restricted site in the lateral expansion of the cartilage and then progressed to other areas of the epiphyseal cartilage. Mineralized structures were identified by von Kossa's staining and by in vivo incorporation of calcein green. Element analysis showed that calcium crystals consisted of poorly crystalline hydroxyapatite. Mineralized matrix was initially spherical structures that generally coalesced after a certain size to occupy larger areas of the cartilage. Alkaline phosphatase activity was detected at the plasma membrane of nearby chondrocytes and in extracellular matrix. Apoptosis was detected by the TUNEL (TDT-mediated dUTP-biotin nick end-labeling) reaction in some articular chondrocytes from mineralized areas. The area occupied by calcium crystals increased significantly in older animals, especially in areas under compression. Ultrastructural analyses showed clusters of needle-like crystals in the extracellular matrix around the chondrocytes and large blocks of mineralized matrix. In 4-year-old animals, some lamellar bone (containing bone marrow) occurred in the same area as articular cartilage mineralization. These results show that the articular cartilage of R. catesbeiana undergoes precocious and progressive mineralization that is apparently stimulated by compressive forces. We suggest that this mineralization is involved in the closure of bone extremities, since mineralization appears to precede the formation of a rudimentary secondary center of ossification in older animals.

Testicular morphological changes during the seasonal reproductive cycle in the bullfrog Rana catesbeiana

Spermatogenesis and steroidogenesis undergo seasonal variations during the reproductive cycle in amphibians. Testicular morphological and morphometric seasonal variations as well as interstitial lipidic inclusions and intralobular glycoconjugates were evaluated during seasonal cycle of Rana catesbeiana. Testes of frogs collected during the annual seasons were weighed for calculation of GSI (Gonadosomatic index). Seminiferous lobule diameters (DSL) and volume densities of seminiferous lobules (VvSL), excretory ducts (VvED), and interstitial tissue (VvIT) were analyzed. Semithin sections were submitted to Periodic Acid-Schiff (PAS) and Alcian Blue (AB) methods for detection of glycoconjugates, while lipidic inclusions were detected by Sudan Black B. GSI showed no significant variations during the year. Since VvED and VvIT increased significantly during summer and were inversely proportional to VvSL, a compensatory effect between the testicular compartments may be related to the maintenance of GSI. During autumn/winter, larger lobular diameters were observed in comparison to spring/summer when spermiogenesis and spermiation were commonly observed. The increased VvIT and the numerous lipidic inclusions in the interstitial cells during summer suggest a relationship between spermiogenesis and steroidogenesis. Besides the structural stability variations occurring in the IT and SL, a possible paracrine interaction between ED and IT should be also involved in the IT development during summer. The presence of PAS and AB-positive globular structures were observed in the seminiferous lobules and excretory ducts. These structures containing acid glycoconjugates appear to be Sertoli cell apical portions, which are accumulated in the lumen of the seminiferous lobules mainly during spermiation. © 2004 Wiley-Liss, Inc.

Evaluation of bullfrog, rana catesbeiana, management in Illinois : a report to the Illinois Department of Conservation through the Illinois Natural History Survey /

Prepared by Philip C. Mankin, Jeffery L. Van Orman, Bruce D. Taubert, David P. Philipp.

Inclusão de farinha de resíduos da indústria de filetagem de tilápias na alimentação de girinos de rã-touro (Rana Catesbeiana Shaw,1802)

The objective of this experiment was to evaluate the inclusion of meal of tilapia industrial waster in bullfrog tadpoles' diets (Rana Catesbeina Shaw, 1802). Two hundred fifty tadpoles with initial weight and length of 0,18 +/- 0,05g and 2,50 +/- 0,18 centimeters were used, respectively, they were used and distributed in 25 aquariums of 30 liter each in a complete randomized design with five treatments and five repetitions. The tadpoles were fed four times a day with diets containing 0, 5, 10 and 15% of inclusion of tilapia filleting and another diet with 0% of supplemented tilapia filleting with synthetic methionine, being supplied four times a day at ease. The experiment period was 30 days. The diets with tilapia filleting inclusion and methionine supplementation did not have influence in the final weight and length averages, feed survival and conversion of bullfrog tadpoles. It cam be used until 15% of tilapia filleting inclusion in the bullfrog tadpoles food. The synthetic methionine supplementation in diets for bullfrog tadpoles is not necessary in diets with levels over 0,471/4 of the total methionine.

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.

Suplementação com vitamina C e E sobre o desenvolvimento e sobrevivência de girinos de rã-touro (Rana catesbeiana Shaw, 1802)

O presente trabalho teve como objetivo avaliar os efeitos da suplementação de vitamina C e E sobre o desenvolvimento e sobrevivência de girinos de rã-touro. Os animais receberam 50, 250 e 500mg de vitamina C ou E/kg de ração, distribuídos segundo delineamento inteiramente casualizado com 3 repetições cada. Na dieta controle, não foram adicionadas vitaminas. Os girinos suplementados com vitamina C ou E não apresentaram diferença significativa no crescimento, comparados ao controle. Não obstante, os suplementados com 500mg de vitamina C apresentaram melhor sobrevivência (62,83%) do que os animais controle (40,48%; p<0,05), nos demais níveis desta vitamina e a adição de vitamina E não alteraram a sobrevivência. O presente trabalho sugere que 500mg de vitamina C/kg de ração constitui uma opção para melhorar a sobrevivência de girinos de rã-touro.

Influência de probióticos no desempenho produtivo e fisiológico de rã-touro Rana catesbeiana Shaw, 1802

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)