Physicochemical, Microbiological, and Toxicological Characterization of Pâté Prepared from the Meat and Liver of Bullfrog (Aquarana catesbeiana) Carcasses.

The development of balanced, healthy, ready-to-consume, and easy-to-prepare products has led to the development of new food technologies. Despite their high commercial value, bullfrog (Aquarana catesbeiana) carcasses result in low yields, with the thighs being the most marketed in comparison to other carcass portions. In this sense, liver pâté is a traditional food consumed worldwide, mainly in European countries, and may be prepared by incorporating bullfrog meat by-products and certain viscera. In this context, the aim of the present study was to develop a pâté product based on a mixture comprising 50% grounded bullfrog torso meat and 50% liver paste, with each treatment incorporating 10% liver paste increments, totaling five final mixtures. The nutritional compositions and physicochemical, microbiological, and toxicological characteristics of each mixture were assessed. The dry matter percentage of the prepared product was determined to be 27.00%, while mineral content was 1.45%, lipid content was 4.00%, and total protein content was 20.00%. Finally, microbiological counts were in agreement with current food safety regulations. The developed pâté serves as a standard, recycling underused industrial materials, adding value to the production chain at low operational costs, creating a more accessible market, and promoting the popularization of this type of meat.

Contrasting effects of transdermal and implant corticosterone treatments in the American bullfrog wound healing.

Glucocorticoid (GC) release is triggered by adverse stimuli that activate the hypothalamus-pituitary-adrenal/interrenal axis. Glucocorticoids may enhance or suppress immune functions depending on the level of elevation. In this study, we investigated the effects of transient and chronic increase of corticosterone (CORT) on the wound healing of the American bullfrog. Frogs were submitted to a daily transdermal hormonal application that acutely elevated CORT plasma levels, or vehicle as a control. Other frogs were surgically implanted with a silastic tube filled with CORT that resulted in chronic elevation of CORT plasma levels or received empty implants as a control. A dermal biopsy was performed to create a wound and was photographed every 3 days. Individuals treated with transdermal CORT started healing faster than their control 32 days after the biopsy. Frogs that received CORT implants tended to heal slower than control subjects. Plasma bacterial killing ability was not affected by treatment, which reinforces the constitutive nature of this innate immune trait. By the end of the experiment, frogs from the acute CORT treatment had smaller wounds compared with those receiving the CORT-filled implants, highlighting the differential effects of acute (immunoenhancing) and chronic (immunosuppressive) elevation of CORT plasma levels. This article is part of the theme issue 'Amphibian immunity: stress, disease and ecoimmunology'.

First evidence of ranavirus in native and invasive amphibians in Colombia.

Ranaviruses can cause mass mortality events in amphibians, thereby becoming a threat to populations that are already facing dramatic declines. Ranaviruses affect all life stages and persist in multiple amphibian hosts. The detrimental effects of ranavirus infections to amphibian populations have already been observed in the UK and in North America. In Central and South America, the virus has been reported in several countries, but the presence of the genus Ranavirus (Rv) in Colombia is unknown. To help fill this knowledge gap, we surveyed for Rv in 60 species of frogs (including one invasive species) in Colombia. We also tested for co-infection with Batrachochytrium dendrobatidis (Bd) in a subset of individuals. For Rv, we sampled 274 vouchered liver tissue samples collected between 2014 and 2019 from 41 localities covering lowlands to mountaintop páramo habitat across the country. Using quantitative polymerase chain reaction (qPCR) and end-point PCR, we detected Rv in 14 individuals from 8 localities, representing 6 species, including 5 native frogs of the genera Osornophryne, Pristimantis and Leptodactylus, and the invasive American bullfrog Rana catesbeiana. Bd was detected in 7 of 140 individuals, with 1 co-infection of Rv and Bd in an R. catesbeiana specimen collected in 2018. This constitutes the first report of ranavirus in Colombia and should set off alarms about this new emerging threat to amphibian populations in the country. Our findings provide some preliminary clues about how and when Rv may have spread and contribute to understanding how the pathogen is distributed globally.

Gastrointestinal parasites in bullfrogs (Lithobates catesbeianus) in aquaculture production units in the Mexican central highlands / Parasitos gastrointestinais em rãs-touro (Lithobates catesbeianus) em unidades de produção aquícola do planalto central do México

In Mexico, intensive production of bullfrogs is one of the most important aquaculture activities, due to growing demand for their meat. Frogs can be hosts for several parasites that negatively affect their development and health. The objective of this study was to identify the presence of intestinal parasites in bullfrogs in aquaculture production units. Eighteen bullfrogs aquaculture production units were selected, and 20 animals (n=360) from each farm. Fecal samples were obtained by mucosal scraping and processed using the concentration method. The overall prevalence of intestinal parasites was 70.5%, and all farms had frogs infected by some species of parasite. Two species of parasites were identified: Eimeria sp. and Strongyloides sp. Significant differences were found regarding parasite prevalence between males and females (73.8% vs 58.8%) and regarding tibia length (5.5 vs 6.1 cm) and weight (168 vs 187 g) between parasitized and non-parasitized frogs. In conclusion, the present study showed a high prevalence of intestinal parasites, and morphometric alterations (weight, snout-cloaca length, radio-ulna length, tibia length and distance between parotid glands) were identified in the parasitized animals. These results provided useful information that will enable establishment of adequate control measures to help minimize the adverse effects of these parasites.(AU)

No México, a produção intensiva de rãs-touro é uma das atividades mais relevantes da aquicultura devido à sua crescente demanda. As rãs podem ser hospedeiras de vários parasitos que afetam negativamente seu desenvolvimento e sua saúde animal. O objetivo deste trabalho é identificar a presença de parasitas gastrointestinais (IPs) em rãs de boi em unidades de produção aquícola. Foram selecionadas 18 unidades de produção aquícola de rãs-touro, e de cada unidade 20 animais (n=360). Amostras fecais foram obtidas por raspagem de mucosas e processadas pelo método de concentração. A prevalência geral de IPs foi de 70,5%, todas as fazendas foram infectadas com algum parasita. Duas espécies de parasitas Eimeria sp. e Strongyloides sp. foram identificadas; encontrando-se diferenças significativas na prevalência entre machos e fêmeas (73,8% vs 58,8%), comprimento da tíbia (5,5 vs 6,1 cm) e peso (168 vs 187 g) entre rãs parasitadas e não parasitadas. Em conclusão, o presente estudo mostrou uma alta prevalência e alterações morfométricas (peso, comprimento do ronco-cloaca, comprimento do rádio-ulna, comprimento da tíbia e distância entre as glândulas parótidas) todas identificadas nos animais parasitados. Esses resultados mostram informações relevantes que permitirão o estabelecimento de medidas de controle adequadas para ajudar a minimizar os efeitos adversos desses parasitas.(AU)

Productive diagnosis offrog culture in the state of Rio de Janeiro / Diagnóstico produtivo da ranicultura no estado do Rio de Janeiro

Brazilian frog farming has a history of production that began in the state of Rio de Janeiro in 1935. Over the years, this activity has spread throughout Brazil, with technological advances that have improved the productivity and health of enterprises. However, structuring the production chain has presented moments of growth and decline, culminating in low production compared with other aquaculture activities. Despite efforts focused on restructuring the chain, data on enterprises and their actors remain scarce. To obtain information on the current scenario of frog farming in the state of Rio de Janeiro, in this study, an online survey of data on frog producers in Rio de Janeiro was conducted. In general, frog farming in Rio de Janeiro has not shown substantial improvements in terms of structuring. Its dynamics are still linked to lowproduction, family run enterprises, little insertion into the market, and not being formalized, encompassing aspects related to fiscal, environmental, and health issues. Therefore, the restructuring process of the frog chain in the state depends on overcoming the bottlenecks mentioned in this study. These predominantly refer to the availability of information on the relevant aspects of management and regularization of production.(AU)

A ranicultura brasileira apresenta um histórico de produção que se iniciou no estado do Rio de Janeiro em 1935. Com o passar dos anos essa atividade se difundiu para todo o Brasil, apresentando avanços tecnológicos que melhoraram o manejo sanitário e a produtividade dos empreendimentos. Contudo, a estruturação da cadeia produtiva apresentou momentos de crescimento e declínio, culminando com uma baixa produção em comparação com outras atividades aquícolas. Apesar dos esforços focados na reestruturação da cadeia, ainda são escassos dados sobre os empreendimentos e seus atores. Com o objetivo de obter informações acerca do cenário atual da ranicultura no estado do Rio de Janeiro, o presente trabalho executou levantamento de dados referentes aos produtores de rã fluminense por meio de formulário online. De uma maneira geral, a ranicultura no Rio de Janeiro não conseguiu apresentar grandes melhorias em termos de estruturação. Sua dinâmica ainda está atrelada a empreendimentos com baixa produção, familiar, pouca inserção no mercado e não formalizados, englobando aspectos relacionados às questões fiscais, ambientais e sanitárias. Assim, o processo de reestruturação da cadeia ranícola no estado depende da superação dos gargalos apontados neste estudo, principalmente referentes à disponibilização de informação acerca dos aspectos de manejo e regularização da produção.(AU)

Evaluation of nutritional composition and technological functionality of whole American Bullfrog (Lithobates catesbeianus), its skin, and its legs as potential food ingredients.

Frog farming systems do not take advantage of their byproducts, which represents health risks and environmental pollution. The present study aimed to evaluate the proximate composition, amino acid, and fatty acid profile of American Bullfrog byproducts (whole frogs (WF), legs (LF), and skin (SF)) and their technological functionality. Results showed that WF, LF, and SF protein content was 47.6, 88.4, and 91.1% dry base (d.b.), correspondingly. Fat content resulted in 34.6, 3.2, and 4.2% (d.b.), respectively. Moreover, byproducts contain all the essential amino acids (23.8-46.6%), and the unsaturated fatty acids predominated the saturated fats. Samples showed water and oil absorption capacities of 1.8-2.6% and 1.8-4.0%, respectively, while oil and water emulsion capacities were 76.7-98.3% and 36.1-85.6%, correspondingly. Additionally, SF presented a gelling capacity in a 5% concentration. These results showed that frogs' byproducts have adequate nutritional and functional capacities, compared to other vegetable and animal flours used in the industry.

Anti-Inflammatory Activity of Bullfrog Oil Polymeric Nanocapsules: From the Design to Preclinical Trials.

BACKGROUND: Although bullfrog oil (BFO) exerts anti-inflammatory effects, it has undesirable properties limiting its use. METHODOLOGY: BFO nanocapsules (BFONc) were produced through nanoprecipitation, and their physicochemical and morphological properties were characterized. To evaluate the biocompatibility of the formulation, a mitochondrial activity evaluation assay was conducted, and cell uptake was assessed. The in vitro anti-inflammatory activity was evaluated by measuring reactive oxygen species (ROS), nitric oxide (NO), type-6 interleukin (IL-6), and tumor necrosis factor (TNF) levels. The in vivo anti-inflammatory effect was assessed by quantifying myeloperoxidase (MPO) levels using the carrageenan-induced paw edema model. RESULTS: BFONc showed a particle size of 233 ± 22 nm, a polydispersity index of 0.17 ± 0.03, and a zeta potential of -34 ± 2.6mV. BFONc revealed remarkable biocompatibility and did not induce changes in cell morphology. Furthermore, BFONc decreased ROS levels by 81 ± 4%; however, NO level increased by 72 ± 18%. TNF and IL-6 levels were reduced by approximately 10% and 90%, respectively. Significant in vivo anti-inflammatory activity was observed compared to dexamethasone. MPO levels were reduced up to 2 MPOs/mg. CONCLUSION: Taken together, the results pointed out the remarkable biocompatibility and anti-inflammatory effects of BFONc.

Orexin-A inhibits fictive air breathing responses to respiratory stimuli in the bullfrog tadpole (Lithobates catesbeianus).

In pre-metamorphic tadpoles, the neural network generating lung ventilation is present but actively inhibited; the mechanisms leading to the onset of air breathing are not well understood. Orexin (ORX) is a hypothalamic neuropeptide that regulates several homeostatic functions, including breathing. While ORX has limited effects on breathing at rest, it potentiates reflexive responses to respiratory stimuli mainly via ORX receptor 1 (OX1R). Here, we tested the hypothesis that OX1Rs facilitate the expression of the motor command associated with air breathing in pre-metamorphic bullfrog tadpoles (Lithobates catesbeianus). To do so, we used an isolated diencephalic brainstem preparation to determine the contributions of OX1Rs to respiratory motor output during baseline breathing, hypercapnia and hypoxia. A selective OX1R antagonist (SB-334867; 5-25 µmol l-1) or agonist (ORX-A; 200 nmol l-1 to 1 µmol l-1) was added to the superfusion media. Experiments were performed under basal conditions (media equilibrated with 98.2% O2 and 1.8% CO2), hypercapnia (5% CO2) or hypoxia (5-7% O2). Under resting conditions gill, but not lung, motor output was enhanced by the OX1R antagonist and ORX-A. Hypercapnia alone did not stimulate respiratory motor output, but its combination with SB-334867 increased lung burst frequency and amplitude, lung burst episodes, and the number of bursts per episode. Hypoxia alone increased lung burst frequency and its combination with SB-334867 enhanced this effect. Inactivation of OX1Rs during hypoxia also increased gill burst amplitude, but not frequency. In contrast with our initial hypothesis, we conclude that ORX neurons provide inhibitory modulation of the CO2 and O2 chemoreflexes in pre-metamorphic tadpoles.

Metabolic, immunologic, and histopathologic responses on premetamorphic American bullfrog (Lithobates catesbeianus) following exposure to lithium and selenium.

The presence of chemicals and the destruction of freshwater habitats have been addressed as one of the reasons for the decline in the amphibians' populations worldwide. Considering the threat that these animals have been suffering in tropical regions, the present study tested if the Brazilian legislation, concerning the permissive levels of lithium and selenium in water bodies and effluents, warrants the protection of aquatic life. To do so, we assessed the metabolic, immunologic, and histopathologic alterations in liver samples of American bullfrog (Lithobates catesbeianus), at the premetamorphic stage, through biomarkers indicative of general energetic status, i.e., glucose, lipid, and protein metabolism using biochemical and histochemical approaches. The immunologic responses were assessed by the quantification of melanomacrophage centres (MMCs); the histopathologic evaluation of the liver sections was also performed. The assay was carried out over 21 days with two periods of sampling (after 7 and 21 days) to assess the effects of exposure over time. The animals were exposed to the considered safe levels of lithium (2.5 mg L-1) and selenium (10 µg L-1), both, isolated and mixed. The exposed animals showed alterations in glucose and lipid metabolism throughout the experiment. The intense presence of MMCs and histopathological responses are compatible with hepatotoxicity. The toxicity expressed by the employed animal model indicates that the Brazilian environmental legislation for the protection of aquatic life needs to be updated. With this study, we intend to provide data for better environmental policies and bring attention to sublethal effects triggered by the presence of contaminants in the aquatic environment.

Assessment of antioxidant system, cholinesterase activity and histopathology in Lithobates catesbeianus tadpoles exposed to water from an urban stream.

Anthropogenic activities promote changes in community structure and decrease the species abundance of amphibians. The aim of this study was to assess potential alterations in the antioxidant system and cholinesterase activity, histopathological and oxidative damage in Lithobates catesbeianus tadpoles exposed to water from the Cascavel River, in Southern Brazil. Water samples (140 L each) were collected from the headwater, urban and rural areas of the river. Tadpoles were acclimated for seven days. After acclimatization tadpoles were reared in water from the river, except for the control aquarium. After seven days, a portion of the liver was removed and prepared for cholinesterase (ChE), superoxide dismutase (SOD), catalase (CAT) and lipid peroxidation (LPO) analysis; another part of the tissue was prepared for histological examination. An elevation of CAT activity was observed for water from both urban and rural environments. A decrease in LPO reaction was detected, mainly among the tadpoles exposed to water from the rural area. These alternations can cause delay the metamorphosis and lead to metabolic dysfunction, interfering in survival capacity and diminishing, not only individual fitness, but that of the whole population.

Therapeutic bullfrog oil-based nanoemulsion for oral application: Development, characterization and stability.

The aim of this study was to develop, optimize, and characterize a stable therapeutic bullfrog oil based nanoemulsion for oral application using a rational experimental design approach. The optimized oral nanoemulsion contained 0.2 % sodium benzoate and 0.02 % propyl-paraben as preservatives; 0.1 % sucralose and 0.4 % acesulfam K as sweeteners and 0.1 % tutti-frutti as flavoring to mask the unpleasant organoleptic characteristics of bullfrog oil. The oral O/W-nanoemulsion showed the droplet size, PDI, zeta potential, and pH of 410 ± 8 nm, 0.20 ± 0.02, -38 ± 2.5 mV, and 6.43 ± 0.05, respectively. The optimized oral nanoemulsion showed a milky single-phase and optimal physical stability at 25 °C for 90 days. Indeed, higher oxidation induction time and lower formation of peroxides in the oral nanoemulsion were responsible for improving its stability. A therapeutic delivery system containing bullfrog oil for oral application was successfully developed and optimized with ideal thermo-oxidative stability.

Effect of parasitism of Lernaea cyprinacea on tadpoles of the invasive species Lithobates catesbeianus.

The introduction of invasive species is one of the greatest threats currently faced by natural ecosystems, causing ecological imbalances between native populations and transmission of a variety of diseases. We reported the interaction between two exotic species given by the parasitic infestation of the copepod Lernaea cyprinacea in the early stages of the development of the American bullfrog Lithobates catesbeianus in the central area of Argentina. In this paper we analysed the leukocyte profile of parasitized and non-parasitized tadpoles of L. catesbeianus with L. cyprinacea and their body condition (BC) as biomarkers of the health status of organisms. A total of 27 tadpoles of L. catesbeianus were analysed (12 non-parasitized and 15 parasitized). The lower BC recorded in parasitized organisms show a lower health status in these tadpoles, which could be affecting the metamorphosis and therefore impact at the population level. Leukocyte response of L. catesbeianus tadpoles to the parasitism of L. cyprinacea was found. Mature and immature lymphocyte frequencies and hematocrit were higher in parasitized compared to non-parasitized tadpoles, which is a typical response to the presence of parasites. However, eosinophils and monocytes were recorded at high frequencies in not parasitized tadpoles, which could be due to the important role played by these leucocytes in the metamorphosis of frogs. The results of this study constitute a first antecedent on leukocyte profile in aquatic stages of anurans during an ectoparasitosis and its possible implications for environmental health. The parasitism of L. cyprinacea influences the biology of the American bullfrog at both the individual and population levels. Parasitized individuals are not killed directly by the parasite, but they can create conditions for secondary infections, growth retardation, behavioral changes and, ultimately, reduce populations.

Bullfrog oil (Rana catesbeiana Shaw) induces apoptosis, in A2058 human melanoma cells by mitochondrial dysfunction triggered by oxidative stress.

Bullfrog oil, an animal oil extracted from the adipose tissue of Rana catesbeiana Shaw, showed promising cytotoxic activity against melanoma cells and, therefore, has the potential to become a pharmaceutical active compound. However, there is a lack of information regarding the pathways involved in its pharmacological activity. Thus, the aim of this study was to investigate and elucidate the cytotoxic effect of this oil against A2058 human melanoma cells. The cytotoxic potential was evaluated by the MTT assay, the cell cycle analysis and the cell death assay. In addition, the apoptotic potential was investigated by (i) the DNA fragmentation using propidium iodide staining analysis, (ii) the evaluation of mitochondrial membrane potential and (iii) the determination of intracellular Reactive Oxygen Species (ROS) level. The results showed that the bullfrog oil was able to promote a time-dependent cytotoxic effect, decreasing cell viability to 38% after 72 h of treatment without affecting the cell cycle. Additionally, the bullfrog oil induced the apoptosis in A2058 cells, increasing up to 50 ±â€¯13% of the intracellular ROS level, maintaining the DNA integrity and promoting an approximate decrease of 35 ±â€¯5% in the mitochondrial membrane potential. It can be concluded that the in vitro cytotoxic effect of the bullfrog oil in A2058 human melanoma cells is mediated by oxidative stress that induces mitochondrial dysfunction, triggering the apoptosis. These unprecedented results highlight the pharmacological potential of bullfrog oil and provide important information to support studies on the development of new pharmaceutical products for complementary and alternative treatments for melanoma.

Discovery of Wild Amphibians Infected with Ranavirus in Brazil.

Ranavirus is a double-stranded DNA virus associated with amphibian, fish and reptile die-offs worldwide. International trade of live animals farmed for human consumption, such as the American bullfrog (Lithobates catesbeianus), plays a key role in spreading the pathogen. In Brazil, ranavirus has only been reported in captive bullfrog farms. We found infected tadpoles of both native species and the American bullfrog in the wild, and a case of mass mortality of amphibians and fish potentially associated with ranavirus. Dead animals presented skin ulcerations, hemorrhages, and edemas. We also found an overall prevalence of 37% of the amphibian chytrid in the area, and two bullfrog tadpoles were co-infected with both pathogens. We suggest that the interaction between the two pathogens should be investigated to improve global conservation of ectothermic vertebrates.

Buccal Bullfrog (Rana catesbeiana Shaw) Oil Emulsion: A Mucoadhesive System Intended for Treatment of Oral Candidiasis.

Oral candidiasis (OC) is an infectious disease caused by microorganisms of the genus Candida, leading to lesions in the buccal cavity. Its treatment consists of the administration of topical or systemic antifungal agents, which may compromise the patient compliance due to its side effects, highlighting the need for alternative treatments. In this scenario, bullfrog oil, an animal oil composed of a pool of saturated and unsaturated fatty acids, is introduced as a potential antifungal raw material. Thus, the aim of this work was to produce a mucoadhesive emulsified system able to deliver the bullfrog oil in the buccal cavity to treat the OC. The emulsion was produced and characterized by visual inspection, droplet size, polydispersity index (PdI), and zeta potential over the course of 60 days. In addition, its mucoadhesive ability was evaluated using an in vitro mucin model. The antifungal activity, evaluated by the broth microdilution assay and the biocompatibility, performed against human erythrocytes, were also carried out. The emulsion showed a droplet size of 320.79 ± 35.60 nm, a PdI of 0.49 ± 0.08, and a zeta potential of -38.53 ± 6.23 mV, with no significant changes over 60 days. The mucoadhesive properties of the system was improved by the use of pharmaceutical excipients. The antifungal activity showed that the bullfrog oil and the emulsion were able to inhibit the growth of different Candida species. Furthermore, the emulsion showed no significant hemolytic effect. Overall, the system showed suitable physicochemical characteristics and biocompatibility, with substantial in vitro antifungal activity, suggesting that this system can be further investigated for OC treatment.

Tuberculosis caused by Mycobacterium bovis infection in a captive-bred American bullfrog (Lithobates catesbeiana).

BACKGROUND: Tuberculosis is widely known as a progressive disease that affects endothermic animals, leading to death and/or economical losses, while mycobacterial infections in amphibians are commonly due to nontuberculous mycobacteria. To the authors' knowledge, this report describes the first case of bovine tuberculosis in a poikilothermic animal. CASE PRESENTATION: An adult female captive American bullfrog (Lithobates catesbeianus Shaw, 1802) died in a Brazilian aquarium. Multiple granulomas with acid-fast bacilli were observed in several organs. Identification of Mycobacterium bovis was accomplished by culture and PCR methods. The other animals from the same enclosure were euthanized, but no evidence of mycobacterial infection was observed. CONCLUSIONS: The American bullfrog was introduced in several countries around the world as an alternative husbandry, and its production is purposed for zoological and aquarium collections, biomedical research, education, human consumption and pet market. The present report warns about an episode of bovine tuberculosis in an amphibian, therefore further studies are necessary to define this frog species' role in the epidemiology of M. bovis.

Getting the Jump on the Development of Bullfrog Oil Microemulsions: a Nanocarrier for Amphotericin B Intended for Antifungal Treatment.

Amphotericin B (AmB), a potent antifungal drug, presents physicochemical characteristics that impair the development of suitable dosage forms. In order to overcome the AmB insolubility, several lipid carriers such as microemulsions have been developed. In this context, the bullfrog oil stands out as an eligible oily phase component, since its cholesterol composition may favor the AmB incorporation. Thus, the aim of this study was to develop a microemulsion based on bullfrog oil containing AmB. Moreover, its thermal stability, antifungal activity, and cytotoxicity in vitro were evaluated. The microemulsion formulation was produced using the pseudo-ternary phase diagram (PTPD) approach and the AmB was incorporated based on the pH variation technique. The antifungal activity was evaluated by determination of minimal inhibitory concentration (MIC) against different species of Candida spp. and Trichosporon asahii. The bullfrog oil microemulsion, stabilized with 16.8% of a surfactant blend, presented an average droplet size of 26.50 ± 0.14 nm and a polydispersity index of 0.167 ± 0.006. This system was able to entrap AmB up to 2 mg mL-1. The use of bullfrog oil as oily phase allowed an improvement of the thermal stability of the system. The MIC assay results revealed a growth inhibition for different strains of Candida spp. and were able to enhance the activity of AmB against T. asahii. The microemulsion was also able to reduce the AmB toxicity. Finally, the developed microemulsion showed to be a suitable system to incorporate AmB, improving the system's thermal stability, increasing the antifungal activity, and reducing the toxicity of this drug.

Lithobates catesbeianus (American Bullfrog) oocytes: a novel heterologous expression system for aquaporins.

Xenopus laevis oocytes are a valuable tool for investigating the function of membrane proteins. However, regulations around the world, specifically in Brazil, render the import of Xenopus laevis frogs impractical, and, in some cases, impossible. Here, as an alternative, we evaluate the usefulness of the North American aquatic bullfrog Lithobates catesebeianus, which is commercially available in Brazil, for the heterologous expression of aquaporin (AQP) proteins. We have developed a method that combines a brief collagenase treatment and mechanical defolliculation for isolating individual oocytes from Lithobates ovaries. We find that they have a similar size, shape, and appearance to Xenopus oocytes and can tolerate and survive following injections with cRNA or water. Furthermore, surface biotinylation, western blot analysis, and measurements of osmotic water permeability (Pf) show that Lithobates oocytes can express AQPs to the plasma membrane and significantly increase the Pf of the oocytes. In fact, the Pf values are similar to historical values gathered from Xenopus oocytes. Due to the presence of a mercury sensitive cysteine (Cys or C) in the throat of the water channel, the Pf of oocytes expressing human (h) AQP1, hAQP1FLAG [FLAG, short protein tag (DYKDDDDK) added to the N-terminus of AQP1], hAQP8, and rat (r) AQP9 was inhibited with the mercurial compound p-chloromercuribenzene sulfonate (pCMBS), whereas AQPs lacking this Cys - hAQP1C189S mutant [residue Cys 189 was replaced by a serine (Ser or S)] and hAQP7 - were mercury insensitive. Contrary to previous studies with Xenopus oocytes, rAQP3 was also found to be insensitive to mercury, which is consistent with the mercury-sensitive Cys (Cys 11) being located intracellularly. Thus, we consider Lithobates oocytes to be a readily accessible system for the functional expression and study of membrane proteins for international researchers who do not currently have access to Xenopus oocytes.

Frog tail: a source of protein to feed the future / Cauda de rã: uma fonte proteica para alimentar o futuro

The aim of this study was to produce a nutritious food, in short-time period, of final low cost, based in frog matrix. Tadpoles obtained from natural fertilization were reared for 100 days and then transported to industrial processing plants, where two primary products were elaborated, the frog tail fillet (FTF) and the non-edible part (NEP). Two canned preserves were developed from the FTF (frog tail fillet in oil/FTFoil; frog tail fillet in tomato sauce/FTFtomato) and from the NEP a flour was produced (tadpole flour/TF). The five products were subjected to physical-chemical analysis. Those elaborated from the tail presented chemical composition values close to the frog meat, while NEP and TF showed values of protein, lipid and ash compatible with by-products and ingredients normally used in the animal feed industry. The two canned preserves were subjected to acceptance test and purchase intent test, with emphasis on the FTFoil, which obtained an acceptance rate of 77% from the evaluators. The method of food production proposed proved to be satisfactory fully reaching the objectives stated. It also can be used in anti-hunger programs, as well as by human and animal industries, improving the quality of the products developed.

Objetivou-se a produção de um alimento nutritivo, em curto espaço de tempo, de custo final baixo, utilizando-se como matriz a rã. Girinos obtidos a partir de fertilização natural foram criados durante 100 dias e transportados para plantas processadoras, onde foram elaborados dois produtos, o filé da cauda de rã (FTF) e a parte não-comestível (NEP). Duas conservas enlatadas foram desenvolvidas a partir do FTF (filé da cauda de rã em óleo/ FTFoil; filé da cauda de rã com molho de tomate/FTFtomato) e da NEP, uma farinha foi produzida (farinha de girino/TF). Os cinco produtos foram submetidos à análise físico-química. Àqueles elaborados a partir do FTF apresentaram valores de composição química próximos à carne de rã, enquanto que NEP e TF apresentaram valores de proteínas, lipídios e cinzas compatíveis com coprodutos normalmente usados na indústria de alimentação animal. As duas conservas enlatadas foram submetidas a testes de aceitação e intenção de compra, destacando-se o FTFoil, que obteve uma taxa de aceitação de 77%. O método de produção de alimentos proposto mostrou-se satisfatório alcançando plenamente os objetivos estabelecidos. Os produtos propostos podem ser utilizados em programas de combate à fome, bem como pelas indústrias de alimentos, nas mais diversas formas.

Frog tail: a source of protein to feed the future / Cauda de rã: uma fonte proteica para alimentar o futuro

The aim of this study was to produce a nutritious food, in short-time period, of final low cost, based in frog matrix. Tadpoles obtained from natural fertilization were reared for 100 days and then transported to industrial processing plants, where two primary products were elaborated, the frog tail fillet (FTF) and the non-edible part (NEP). Two canned preserves were developed from the FTF (frog tail fillet in oil/FTFoil; frog tail fillet in tomato sauce/FTFtomato) and from the NEP a flour was produced (tadpole flour/TF). The five products were subjected to physical-chemical analysis. Those elaborated from the tail presented chemical composition values close to the frog meat, while NEP and TF showed values of protein, lipid and ash compatible with by-products and ingredients normally used in the animal feed industry. The two canned preserves were subjected to acceptance test and purchase intent test, with emphasis on the FTFoil, which obtained an acceptance rate of 77% from the evaluators. The method of food production proposed proved to be satisfactory fully reaching the objectives stated. It also can be used in anti-hunger programs, as well as by human and animal industries, improving the quality of the products developed.(AU)

Objetivou-se a produção de um alimento nutritivo, em curto espaço de tempo, de custo final baixo, utilizando-se como matriz a rã. Girinos obtidos a partir de fertilização natural foram criados durante 100 dias e transportados para plantas processadoras, onde foram elaborados dois produtos, o filé da cauda de rã (FTF) e a parte não-comestível (NEP). Duas conservas enlatadas foram desenvolvidas a partir do FTF (filé da cauda de rã em óleo/ FTFoil; filé da cauda de rã com molho de tomate/FTFtomato) e da NEP, uma farinha foi produzida (farinha de girino/TF). Os cinco produtos foram submetidos à análise físico-química. Àqueles elaborados a partir do FTF apresentaram valores de composição química próximos à carne de rã, enquanto que NEP e TF apresentaram valores de proteínas, lipídios e cinzas compatíveis com coprodutos normalmente usados na indústria de alimentação animal. As duas conservas enlatadas foram submetidas a testes de aceitação e intenção de compra, destacando-se o FTFoil, que obteve uma taxa de aceitação de 77%. O método de produção de alimentos proposto mostrou-se satisfatório alcançando plenamente os objetivos estabelecidos. Os produtos propostos podem ser utilizados em programas de combate à fome, bem como pelas indústrias de alimentos, nas mais diversas formas.(AU)