Inclusion complex and nanoclusters of cyclodextrin to increase the solubility and efficacy of albendazole.

Albendazole (ABZ), a benzimidazole widely used to control gastrointestinal parasites, is poorly soluble in water, resulting in variable and incomplete bioavailability. This has favored the appearance ABZ-resistant nematodes and, consequently, an increase in its clinical ineffectiveness. Among the pharmaceutical techniques developed to increase drug efficacy, cyclodextrins (CDs) and other polymers have been extensively used with water-insoluble pharmaceutical drugs to increase their solubility and availability. Our objective was to prepare ABZ formulations, including ß-cyclodextrin (ßCD) or hydroxypropyl-ß-cyclodextrin (HPßCD), associated or not to the water-soluble polymer polyvinylpyrrolidone (PVP). These formulations had their solubility and anthelmintic effect both evaluated in vitro. Also, their anthelmintic efficacy was evaluated in lambs naturally infected with gastrointestinal nematodes (GIN) through the fecal egg count (FEC) reduction test. In vitro, the complex ABZ/HPßCD had higher solubility than ABZ/ßCD. The addition of PVP to the complexes increased solubility and dissolution rates more effectively for ABZ/HPßCD than for ABZ/ßCD. In vivo, 48 lambs naturally infected with GIN were divided into six experimental groups: control, ABZ, ABZ/ßCD, ABZ/ßCD-PVP, ABZ/HPßCD, and ABZ/HPßCD-PVP. Each treated animal received 10 mg/kg of body weight (based on the ABZ dose) for three consecutive days. After 10 days of the last administered dose, treatment efficacy was calculated. The efficacy values were as follows: ABZ (70.33%), ABZ/ßCD (85.33%), ABZ/ßCD-PVP (82.86%), ABZ/HPßCD (78.37%), and ABZ/HPßCD-PVP (43.79%). In vitro, ABZ/HPßCD and ABZ/HPßCD-PVP had high solubility and dissolution rates. In vivo, although the efficacies of ABZ/ßCD, ABZ/ßCD-PVP, and ABZ/HPßCD increased slightly when compared to pure ABZ, this increase was not significant (P > 0.05).

Desempenho do polvo Octopus vulgaris alimentado com monodieta de mexilhão (Perna perna) / Performance of the Octopus vulgaris octopus fed on mussel (Perna perna) as monodiet

The octopus Octopus cf. vulgaris is a potential species to diversify aquaculture. Due to absence of balanced commercial diet, growth of the O. cf. vulgaris is based on natural diet with local and low-cost inputs. In Brazil, studies on experimental octopus ongrowing are recent and there is little available data. We evaluated the performance, survival and food consumption of O. vulgaris fed on mussel Perna perna for 20 days. Six octopuses with initial weight of 415±12.73g (mean±standard deviation) were divided into two groups (n=3 octopuses/group) according to the diet: MC Group (frozen mussels) and MV Group (live mussels). The Weight Gain of octopuses was 273.33±94.52g and 340.00±26.46g; the Absolute Growth Rate was 13.67±4.73 and 17.00±1.32g.dia-1 and the Specific Growth Rate of 2.95±0.58 and 2.64±0.37%.day-1 to MC and MV groups, respectively. There was no significant difference in performance between groups and the survival rate was 100%. Octopuses well accepted both diets and despite the amount of frozen mussels (129±31) was higher than in live mussels (100±19), there was no significant difference regarding the consumption between groups. Our results demonstrate that the mussel Perna perna can be used frozen or live as monodiet in O. cf. vulgaris ongrowing.(AU)

Desempenho do polvo Octopus vulgaris alimentado com monodieta de mexilhão (Perna perna )

ABSTRACT The octopus Octopus cf. vulgaris is a potential species to diversify aquaculture. Due to absence of balanced commercial diet, growth of the O. cf. vulgaris is based on natural diet with local and low-cost inputs. In Brazil, studies on experimental octopus ongrowing are recent and there is little available data. We evaluated the performance, survival and food consumption of O. vulgaris fed on mussel Perna perna for 20 days. Six octopuses with initial weight of 415±12.73g (mean±standard deviation) were divided into two groups (n=3 octopuses/group) according to the diet: MC Group (frozen mussels) and MV Group (live mussels). The Weight Gain of octopuses was 273.33±94.52g and 340.00±26.46g; the Absolute Growth Rate was 13.67±4.73 and 17.00±1.32g.dia-1 and the Specific Growth Rate of 2.95±0.58 and 2.64±0.37%.day-1 to MC and MV groups, respectively. There was no significant difference in performance between groups and the survival rate was 100%. Octopuses well accepted both diets and despite the amount of frozen mussels (129±31) was higher than in live mussels (100±19), there was no significant difference regarding the consumption between groups. Our results demonstrate that the mussel Perna perna can be used frozen or live as monodiet in O. cf. vulgaris ongrowing.

Doctors' awareness concerning primary immunodeficiencies in Brazil.

BACKGROUND: PIDs are a heterogeneous group of genetic illnesses, and delay in their diagnosis is thought to be caused by a lack of awareness among physicians concerning PIDs. The latter is what we aimed to evaluate in Brazil. METHODS: Physicians working at general hospitals all over the country were asked to complete a 14-item questionnaire. One of the questions described 25 clinical situations that could be associated with PIDs and a score was created based on percentages of appropriate answers. RESULTS: A total of 4026 physicians participated in the study: 1628 paediatricians (40.4%), 1436 clinicians (35.7%), and 962 surgeons (23.9%). About 67% of the physicians had learned about PIDs in medical school or residency training, 84.6% evaluated patients who frequently took antibiotics, but only 40.3% of them participated in the immunological evaluation of these patients. Seventy-seven percent of the participating physicians were not familiar with the warning signs for PIDs. The mean score of correct answers for the 25 clinical situations was 48.08% (±16.06). Only 18.3% of the paediatricians, 7.4% of the clinicians, and 5.8% of the surgeons answered at least 2/3 of these situations appropriately. CONCLUSIONS: There is a lack of medical awareness concerning PIDs, even among paediatricians, who have been targeted with PID educational programmes in recent years in Brazil. An increase in awareness with regard to these disorders within the medical community is an important step towards improving recognition and treatment of PIDs.

Growth and survival of the pearl oyster Pteria hirundo (L.,1758) in an intermediate stage of culture in Santa Catarina, Brazil.

There are no records in Brazil for the culture of Pteridae family oysters, genus Pteria and Pinctada. The intermediate culture can be considered one of the critical stages during the oyster farming. The changes in life conditions from a semi-closed environment provided by the collector to the structures utilized for the growth represents a considerable stress factor to which the animals are subjected. The goal of the present study was to evaluate the growth and survival of the pearl oyster Pteria hirundo (Linnaeus, 1758) kept in intermediate lanterns. Seeds of P. hirundo, obtained through larviculture carried out at the Marine Molluscs Laboratory (UFSC), southern Brazil were detached from the collectors, transferred and kept in an intermediate culture system (oyster lantern). During 6 months, growth (dorso-ventral height - DVH in mm) and survival (%) were evaluated. Two size classes were tested: medium (M), between 3.4 and 4.4 mm, and large (L), bigger than 4.5 mm. The density of occupation in each floor of the lantern was 50%. At the end of the experiment, the survivals observed were 90 and 94% for the sizes M and L, respectively. Medium juveniles of Pteria hirundo had an average (Std) increase of 16.1 (±4.38) mm in the dorso-ventral height, and large juveniles an average increase of 11.4 (±3.77) mm after six months of experiment. The results observed during the intermediate stage of cultivation indicate that Pteria hirundo presents survival and growth similar to other species of pearl oysters traditionally used in the pearl industry in several countries.

Evaluation of Cymbopogon schoenanthus essential oil in lambs experimentally infected with Haemonchus contortus.

Hematophagous gastrointestinal parasites cause significant economic losses in small ruminant grazing systems. The growing reports of multi-drug resistant parasites call for intensive research on alternative treatments for anthelmintics to help small ruminants cope with these parasites. Two-month-old lambs with mean body weight (BW) of 22.5 kg were experimentally infected with a multidrug-resistant Haemonchus contortus strain. Infected animals were dosed orally with Cymbopogon schoenanthus essential oil to evaluate its anthelmintic potential. Eighteen animals were allocated into three groups of six animals, and each received one of the following treatments: Group 1 - control (10 mL of water), Group 2 - C. schoenanthus essential oil (180 mg/kg BW); and Group 3 - C. schoenanthus essential oil (360 mg/kg BW). Animals received the oil once a day for 3 consecutive days. Lambs were evaluated clinically for blood biochemistry before, at 1, 5, 10, 15 and 20 days after treatment, and then were euthanized to assess the total worm burden. No statistically significant reduction in fecal egg count, packed cell volume or total worm count was observed after treatments. Also, no statistical difference among group means for blood levels of urea, creatinine, albumin, alkaline phosphatase, aspartate aminotransferase and gamma glutamyl transferase was found. Larval development assay (LDA) and egg hatch assay (EHA) were performed from feces of treated animals at 1, 5, 10 and 15 days after essential oil administration. An inhibition in LDA was observed 1 day after the 3-day treatment in larvae from feces of animals treated with 360 mg/kg essential oil. In conclusion, the essential oil at the doses of 180 mg/kg and 360 mg/kg was safe to sheep, but failed as an anthelmintic treatment when applied to young sheep artificially infected with a multidrug-resistant H. contortus strain.

Anthelmintic activity of Cymbopogon martinii, Cymbopogon schoenanthus and Mentha piperita essential oils evaluated in four different in vitro tests.

Anthelmintic resistance is a worldwide concern in small ruminant industry and new plant-derived compounds are being studied for their potential use against gastrointestinal nematodes. Mentha piperita, Cymbopogon martinii and Cymbopogon schoenanthus essential oils were evaluated against developmental stages of trichostrongylids from sheep naturally infected (95% Haemonchus contortus and 5% Trichostrogylus spp.) through the egg hatch assay (EHA), larval development assay (LDA), larval feeding inhibition assay (LFIA), and the larval exsheathment assay (LEA). The major constituent of the essential oils, quantified by gas chromatography for M. piperita oil was menthol (42.5%), while for C. martinii and C. schoenanthus the main component was geraniol (81.4% and 62.5%, respectively). In all in vitro tests C. schoenanthus essential oil had the best activity against ovine trichostrongylids followed by C. martini, while M. piperita presented the least activity. Cymbopogon schoenanthus essential oil had LC(50) value of 0.045 mg/ml in EHA, 0.063 mg/ml in LDA, 0.009 mg/ml in LFIA, and 24.66 mg/ml in LEA. The anthelmintic activity of essential oils followed the same pattern in all in vitro tests, suggesting C. schoenanthus essential oil could be an interesting candidate for nematode control, although in vivo studies are necessary to validate the anthelmintic properties of this oil.

Larval settlement and spat recovery rates of the oyster Crassostrea brasiliana (Lamarck, 1819) using different systems to induce metamorphosis.

This study aimed at the assessment, in the laboratory, of the larval settlement and spat recovery rates of oysters of the species Crassostrea brasiliana using plastic collectors, epinephrine (C9H13NO3 C4H6O6) and shell powder in settlement tanks. Polypropylene was used attached to bamboo frames. The material was chosen due to its pliability--that favours the spat detachment. Two experiments were carried out; the first between February and April 2008, and the second between November and December 2008 at the Marine Mussel Laboratory of Santa Catarina Federal University (Laboratório de Moluscos Marinhos da Universidade Federal de Santa Catarina). In the first experiment, the scratched plastic collectors were tested consorting them with shell powder; on the second, the plastic collectors were tested consorted with shell powder, only shell powder and epinephrine as the metamorphosis stimulator. The quantification was carried out of the larvae settled in the plastic collectors, and of the recovery and integrity of the spats after their detachment. The first experiment has shown a recovery rate of 48.83% of the spats in comparison with the D larvae used. From this percentage, 4.9% settled in the plastic collectors and 43.93% in shell powder. The second experiment revealed 55.78% regarding the settled spats in comparison with the total of larvae used (using epinephrine), 78.62% in the treatment with the collector plus shell powder and 58.33% in the treatment only with shell powder. Thus, the use of the collector plus shell powder resulted in a greater spat recovery when compared to the other treatments.

Larval settlement and spat recovery rates of the oyster Crassostrea brasiliana (Lamarck, 1819) using different systems to induce metamorphosis / Taxas de assentamento larval e recuperação de sementes da ostra Crassostrea brasiliana (Lamarck, 1819) com uso de diferentes sistemas de indução à metamorfose

This study aimed at the assessment, in the laboratory, of the larval settlement and spat recovery rates of oysters of the species Crassostrea brasiliana using plastic collectors, epinephrine (C9H13NO3 C4H6O6) and shell powder in settlement tanks. Polypropylene was used attached to bamboo frames. The material was chosen due to its pliability - that favours the spat detachment. Two experiments were carried out; the first between February and April 2008, and the second between November and December 2008 at the Marine Mussel Laboratory of Santa Catarina Federal University (Laboratório de Moluscos Marinhos da Universidade Federal de Santa Catarina). In the first experiment, the scratched plastic collectors were tested consorting them with shell powder; on the second, the plastic collectors were tested consorted with shell powder, only shell powder and epinephrine as the metamorphosis stimulator. The quantification was carried out of the larvae settled in the plastic collectors, and of the recovery and integrity of the spats after their detachment. The first experiment has shown a recovery rate of 48.83 percent of the spats in comparison with the D larvae used. From this percentage, 4.9 percent settled in the plastic collectors and 43.93 percent in shell powder. The second experiment revealed 55.78 percent regarding the settled spats in comparison with the total of larvae used (using epinephrine), 78.62 percent in the treatment with the collector plus shell powder and 58.33 percent in the treatment only with shell powder. Thus, the use of the collector plus shell powder resulted in a greater spat recovery when compared to the other treatments.(AU)

O presente estudo teve como objetivo avaliar a taxa de assentamento larval e recuperação de sementes de ostras da espécie Crassostrea brasiliana, em laboratório, através do uso de coletores plásticos, epinefrina (C9H13NO3 C4H6O6) e pó de concha em tanques de assentamento. Foram utilizados coletores plásticos de polipropileno, presos a armações de bambu. O material foi escolhido devido à boa maleabilidade, o que facilita o destacamento das sementes. Foram realizados dois experimentos, o primeiro entre fevereiro e abril de 2008, e o segundo entre novembro e dezembro de 2008 no Laboratório de Moluscos Marinhos da Universidade Federal de Santa Catarina. No primeiro experimento, testaram-se coletores de plástico arranhado consorciado com pó de concha em um tanque de assentamento, enquanto que no segundo foram utilizados dois tanques de assentamento, um contendo os coletores de plástico consorciado com pó de concha e apenas pó de concha, e, no outro tanque, utilizou-se epinefrina como estimulador da metamorfose. Foi realizada a quantificação das larvas assentadas nos coletores plásticos e a taxa de recuperação e integridade das sementes após o destacamento. No primeiro experimento, recuperaram-se 48,83 por cento de sementes em relação às larvas D utilizadas. Deste percentual, 4,9 por cento assentaram em coletores plásticos e 43,93 por cento em pó de concha. No segundo experimento, a porcentagem de sementes assentadas em relação ao total de larvas utilizadas foi de 55,78 por cento com o uso de epinefrina, 78,62 por cento no tratamento com coletor mais pó de concha e de 58,33 por cento no tratamento só com pó de concha. Assim, verifica-se que o uso de coletor mais pó de concha resulta em maior recuperação de sementes se comparado com os demais tratamentos.(AU)

Produção programada e rastreabilidade de larvas e sementes de moluscos em Santa Catarina / Programmed production and traceability for larvae and seeds of mollusks in Santa Catarina

A produção de moluscos marinhos é diretamente dependente da oferta de sementes. Em alguns casos, como a produção de ostras, em especial a espécie Crassostrea gigas no Brasil, devido a algumas dificuldades como a reprodução e a sobrevivência de larvas no ambiente natural, a obtenção de sementes de qualidade só é possível, até o momento, a partir da produção em laboratório. Esta produção depende da manutenção dos reprodutores em sistemas que permitam o condicionamento e a maturação em laboratório, ao longo do ano. Isso limita a produção de larvas e sementes para as estações do ano em que se pode obter reprodutores maduros. Este trabalho mostra as estratégias de maturação e o controle de larvicultura utilizados no Laboratório de Moluscos Marinhos - Universidade Federal de Santa Catarina (LMM-UFSC), os quais permitem a produção de sementes durante todo o ano, de forma programada, relacionada com a demanda do setor produtivo. Desta forma, foi implementado no LMM um setor de condicionado com sistema de refrigeração de larga escala, o que pode diminuir a temperatura de grandes volumes de água do mar até 14oC, em tanques de 2.000 L, com fluxo contínuo de água e comida durante todo o ano. Neste sistema, é possível manter-se até 2.000 ostras para o estoque de animais já condicionados, e até 1.000 ostras para o condicionamento integral (a partir de ostras previamente induzidas para a desova completa).Com essa estratégia, o processo de maturação integral leva aproximadamente 60 dias, com mais 30 dias para a maturação final dos gametas (com o aumento gradual da temperatura até 21 ou 23oC). Pode-se, assim, manter reprodutores maturados em laboratório, em boas condições de alimentação e temperatura, com estoque de gametas para períodos de até seis meses...

The production of marine mollusks is directly dependent of the seeds offer. In some cases, like oysters production, in particular the specie Crassostrea gigas in Brazil, due to some difficulties as reproduction and larvae survival in natural environment, the obtainment of quality seeds is possible until the moment, only from laboratory production. This production depends on the reproducers maintenance and of the maturation condition of the broodstock, along the year. That limits the larvae and seeds production to seasons of the year in which it can obtain mature reproducers...

Produção programada e rastreabilidade de larvas e sementes de moluscos em Santa Catarina / Programmed production and traceability for larvae and seeds of mollusks in Santa Catarina

A produção de moluscos marinhos é diretamente dependente da oferta de sementes. Em alguns casos, como a produção de ostras, em especial a espécie Crassostrea gigas no Brasil, devido a algumas dificuldades como a reprodução e a sobrevivência de larvas no ambiente natural, a obtenção de sementes de qualidade só é possível, até o momento, a partir da produção em laboratório. Esta produção depende da manutenção dos reprodutores em sistemas que permitam o condicionamento e a maturação em laboratório, ao longo do ano. Isso limita a produção de larvas e sementes para as estações do ano em que se pode obter reprodutores maduros. Este trabalho mostra as estratégias de maturação e o controle de larvicultura utilizados no Laboratório de Moluscos Marinhos - Universidade Federal de Santa Catarina (LMM-UFSC), os quais permitem a produção de sementes durante todo o ano, de forma programada, relacionada com a demanda do setor produtivo. Desta forma, foi implementado no LMM um setor de condicionado com sistema de refrigeração de larga escala, o que pode diminuir a temperatura de grandes volumes de água do mar até 14oC, em tanques de 2.000 L, com fluxo contínuo de água e comida durante todo o ano. Neste sistema, é possível manter-se até 2.000 ostras para o estoque de animais já condicionados, e até 1.000 ostras para o condicionamento integral (a partir de ostras previamente induzidas para a desova completa).Com essa estratégia, o processo de maturação integral leva aproximadamente 60 dias, com mais 30 dias para a maturação final dos gametas (com o aumento gradual da temperatura até 21 ou 23oC). Pode-se, assim, manter reprodutores maturados em laboratório, em boas condições de alimentação e temperatura, com estoque de gametas para períodos de até seis meses...(AU)

The production of marine mollusks is directly dependent of the seeds offer. In some cases, like oysters production, in particular the specie Crassostrea gigas in Brazil, due to some difficulties as reproduction and larvae survival in natural environment, the obtainment of quality seeds is possible until the moment, only from laboratory production. This production depends on the reproducers maintenance and of the maturation condition of the broodstock, along the year. That limits the larvae and seeds production to seasons of the year in which it can obtain mature reproducers...(AU)

Sexual stages of the female portion in the scallop Nodipecten nodosus (Linné, 1758) and astaxanthin quantity in each stage.

This work describes the gametogenic cycle of the scallop Nodipecten nodosus kept in a culture system. To this end, during one year, samples were taken from the broodstocks every 30 days to be submitted to macroscopic and microscopic analyses and to measure the amount of astaxanthin. To perform the microscopic evaluation, 5 micro slices from the median portion of the female part of the gonad were submitted to the pattern methodology for histological analyses with paraffin and HE coloration. The remaining portion of the female gonad was lyophilised to extract and quantify the levels of astaxanthin using HPLC. The microscopic analyses revealed four well defined stages for the reproductive cycle. Analyses of data taken throughout the year indicated preferential spawning periods from December to January and from July to September. The astaxanthin analyses showed higher amounts of this carotenoid during the advanced pre-spawning and the initial spawning periods than during gametogenesis, initial pre-spawning, advanced spawning, and the spent stages. According to these results, it was possible to establish a descriptive table of the sexual stages of the female portion of the gonad and the amount of astaxanthin in the sexual stage of the scallop Nodipecten nodosus.

Sexual stages of the female portion in the scallop Nodipecten nodosus (Linné, 1758) and astaxanthin quantity in each stage / Estágios sexuais da porção feminina da gônada da vieira Nodipecten nodosus (Linné, 1758) e a quantidade de astaxantina em cada estágio

This work describes the gametogenic cycle of the scallop Nodipecten nodosus kept in a culture system. To this end, during one year, samples were taken from the broodstocks every 30 days to be submitted to macroscopic and microscopic analyses and to measure the amount of astaxanthin. To perform the microscopic evaluation, 5 μ slices from the median portion of the female part of the gonad were submitted to the pattern methodology for histological analyses with paraffin and HE coloration. The remaining portion of the female gonad was lyophilised to extract and quantify the levels of astaxanthin using HPLC. The microscopic analyses revealed four well defined stages for the reproductive cycle. Analyses of data taken throughout the year indicated preferential spawning periods from December to January and from July to September. The astaxanthin analyses showed higher amounts of this carotenoid during the advanced pre-spawning and the initial spawning periods than during gametogenesis, initial pre-spawning, advanced spawning, and the spent stages. According to these results, it was possible to establish a descriptive table of the sexual stages of the female portion of the gonad and the amount of astaxanthin in the sexual stage of the scallop Nodipecten nodosus.

Este trabalho descreve o ciclo gametogênico da vieira Nodipecten nodosus mantida em ambiente de cultivo. Para isto, durante um ano, amostras de indivíduos reprodutores foram coletadas a cada 30 dias e submetidas à avaliação macroscópica e microscópica e à quantificação de astaxantina. Para a avaliação microscópica, secções de 5 μ da porção mediana feminina da gônada foram submetidas à metodologia de análise histológica padrão em parafina e coloração HE. O restante da porção feminina da gônada foi liofilizado para extração e quantificação de astaxantina em HPLC. A avaliação microscópica permitiu a descrição de quatro estágios bem definidos para o ciclo reprodutivo. Na análise ao longo do ano, foram observados períodos preferenciais de desova em dezembro e janeiro e de julho a setembro. A análise da quantidade de astaxantina, mostrou, nos estádios de pré-desova avançada e de desova inicial, uma maior quantidade desse carotenoide em comparação aos estádios de gametogênese, pré-desova inicial, desova avançada e repouso. Em função desses resultados, foi possível estabelecer um quadro descritivo dos estágios sexuais da porção feminina da gônada e quantidade de astaxantina em cada estágio sexual da vieira Nodipecten nodosus.

Sexual stages of the female portion in the scallop Nodipecten nodosus (Linné, 1758) and astaxanthin quantity in each stage / Estágios sexuais da porção feminina da gônada da vieira Nodipecten nodosus (Linné, 1758) e a quantidade de astaxantina em cada estágio

This work describes the gametogenic cycle of the scallop Nodipecten nodosus kept in a culture system. To this end, during one year, samples were taken from the broodstocks every 30 days to be submitted to macroscopic and microscopic analyses and to measure the amount of astaxanthin. To perform the microscopic evaluation, 5 μ slices from the median portion of the female part of the gonad were submitted to the pattern methodology for histological analyses with paraffin and HE coloration. The remaining portion of the female gonad was lyophilised to extract and quantify the levels of astaxanthin using HPLC. The microscopic analyses revealed four well defined stages for the reproductive cycle. Analyses of data taken throughout the year indicated preferential spawning periods from December to January and from July to September. The astaxanthin analyses showed higher amounts of this carotenoid during the advanced pre-spawning and the initial spawning periods than during gametogenesis, initial pre-spawning, advanced spawning, and the spent stages. According to these results, it was possible to establish a descriptive table of the sexual stages of the female portion of the gonad and the amount of astaxanthin in the sexual stage of the scallop Nodipecten nodosus.(AU)

Este trabalho descreve o ciclo gametogênico da vieira Nodipecten nodosus mantida em ambiente de cultivo. Para isto, durante um ano, amostras de indivíduos reprodutores foram coletadas a cada 30 dias e submetidas à avaliação macroscópica e microscópica e à quantificação de astaxantina. Para a avaliação microscópica, secções de 5 μ da porção mediana feminina da gônada foram submetidas à metodologia de análise histológica padrão em parafina e coloração HE. O restante da porção feminina da gônada foi liofilizado para extração e quantificação de astaxantina em HPLC. A avaliação microscópica permitiu a descrição de quatro estágios bem definidos para o ciclo reprodutivo. Na análise ao longo do ano, foram observados períodos preferenciais de desova em dezembro e janeiro e de julho a setembro. A análise da quantidade de astaxantina, mostrou, nos estádios de pré-desova avançada e de desova inicial, uma maior quantidade desse carotenoide em comparação aos estádios de gametogênese, pré-desova inicial, desova avançada e repouso. Em função desses resultados, foi possível estabelecer um quadro descritivo dos estágios sexuais da porção feminina da gônada e quantidade de astaxantina em cada estágio sexual da vieira Nodipecten nodosus.(AU)

Carotenoid extraction from the gonad of the scallop Nodipecten nodosus (Linnaeus, 1758) (Bivalvia: Pectinidae).

In marine bivalve mollusks, unsaturated molecules called carotenoids are present in the natural diet and play an important role in different biological process, especially in reproduction. In order to gain more insights into these compounds in Nodipecten nodosus it was necessary to develop a suitable protocol for extraction of carotenoids from the gonads. Female gonads of cultured scallops (75 mm length) were lyophilized and macerated in liquid N2. To verify the effect of composition in organosolvents on the extracting solutions, two organic solvents were tested: acetone and hexane (Ac = O:Hex) at four ratios, 1:1, 1:3, 1:5, and 2:3, in four static extraction times: 0, 5, 10, and 15 minutes. Total carotenoids and astaxanthin contents were determined in the crude extracts by UV-visible spectrophotometry and high performance liquid chromatography (HPLC), respectively. Triplicate aliquots of 50 mg were used for each treatment. The results indicated that the best single extraction (0.312 +/- 0.016 microg carotenoids/mg) was attained with Ac = O: Hex 1:3, for 15 minutes. Through exhaustive extraction methodology (10x), a superior yield (0.41 +/- 0.001 microg carotenoids/mg) was obtained from a gonad sample in comparison to the highest value found for a single extraction. Astaxanthin content was reduced by 8.6% in carotenoid extract preservation assay, i.e., -18 degrees C, 26 days incubation, under N2 atmosphere.

Carotenoid extraction from the gonad of the scallop Nodipecten nodosus (Linnaeus, 1758) (Bivalvia: Pectinidae) / Extração de carotenóides da gônada da vieira Nodipecten nodosus (Bivalvia: Pectinidae)

In marine bivalve mollusks, unsaturated molecules called carotenoids are present in the natural diet and play an important role in different biological process, especially in reproduction. In order to gain more insights into these compounds in Nodipecten nodosus it was necessary to develop a suitable protocol for extraction of carotenoids from the gonads. Female gonads of cultured scallops (75 mm length) were lyophilized and macerated in liquid N2. To verify the effect of composition in organosolvents on the extracting solutions, two organic solvents were tested: acetone and hexane (Ac = O:Hex) at four ratios, 1:1, 1:3, 1:5, and 2:3, in four static extraction times: 0, 5, 10, and 15 minutes. Total carotenoids and astaxanthin contents were determined in the crude extracts by UV-visible spectrophotometry and high performance liquid chromatography (HPLC), respectively. Triplicate aliquots of 50 mg were used for each treatment. The results indicated that the best single extraction (0.312 ± 0.016 µg carotenoids/mg) was attained with Ac = O: Hex 1:3, for 15 minutes. Through exhaustive extraction methodology (10x), a superior yield (0.41 ± 0.001 µg carotenoids/mg) was obtained from a gonad sample in comparison to the highest value found for a single extraction. Astaxanthin content was reduced by 8.6 percent in carotenoid extract preservation assay, i.e., -18 °C, 26 days incubation, under N2 atmosphere.

Em moluscos bivalves marinhos, carotenóides insaturados estão presentes na dieta natural, com um importante papel em diversos processos biológicos, em especial na reprodução. A elucidação dos efeitos destes compostos em Nodipecten nodosus requer o desenvolvimento de um protocolo adequado para a extração de carotenóides das gônadas desses animais. Para isso, gônadas de vieiras cultivadas (75 mm de comprimento) foram liofilizadas e maceradas em N2 líquido. Amostras em triplicata com 50 mg foram coletadas para a utilização em cada tratamento. Os conteúdos de carotenóides totais e astaxantina foram determinados via espectrofotometria de luz UV-visível e cromatografia líquida de alta eficiência (CLAE), respectivamente. O efeito da composição em organosolventes das soluções de extração foi testado utilizando-se acetona (Ac = O) e hexano (Hex) em quatro proporções (Ac = O:Hex): 1:1, 1:3, 1:5, e 2:3; em quatro tempos de extração: 0, 5, 10, e 15 minutos. Os resultados mostraram que o melhor rendimento de extração (0,312 ± 0,016 µg carotenóides/mg) foi obtido com Ac = O:Hex, 1:3, por 15 minutos. Com a utilização de protocolo de extração exaustiva (10x), uma quantidade superior (0,41 ± 0,001 µg de carotenóides/mg) foi obtida de amostras de gônada, comparativamente aos valores obtidos em extrações únicas. O conteúdo de astaxantina foi reduzido em 8,6 por cento em testes de preservação deste metabólito em extratos crus (-18 °C, 26 dias de incubação em atmosfera de N2).

Micronucleus induction in mussels exposed to okadaic acid.

Some toxins present in the marine environment are capable of inducing mutagenicity and/or carcinogenicity. Among these toxins, okadaic acid (OA) is gaining considerable interest since it induces DNA based modifications at low concentrations and accumulates in filter-feeding marine animals, including those used for human consumption. This study aims to evaluate the genotoxicity of OA in the haemocytes of the mussel Perna perna, using the micronucleus assay. Fifty-four mussels were separated into three groups of 18 animals. One group received 0.3 microg of OA diluted in 10 microl of ethanol and ultrapure water while the other groups were considered as controls and were exposed to a solvent plus seawater mixture. A significantly higher frequency of micronuclei was observed in haemocytes from the OA-exposed group. There were no statistical differences between the two control groups.