Combined effects of Limonene and Ivermectin on P-glycoprotein-9 gene expression of lambs Infected with Haemonchus contortus.

Although ivermectin (IVM) has a wide spectrum and long half-life, its frequent use as an anthelmintic for the last 42 years led to its worldwide tolerance by Haemonchus contortus. We evaluated the combination of limonene (LIM), a P-glycoprotein (Pgp) modulator, with IVM in lambs infected with a multidrug-resistant H. contortus. Twenty-four male Dorper lambs were artificially infected with two doses (seven days apart) of 8000 infective larvae of a multidrug-resistant isolate of H. contortus. The infection was patent 25 days later. Fifteen days before treatment with IVM (DAY -15), animals were divided into 4 groups: Infected-untreated control (CTL), IVM, LIM, and LIM+IVM. From DAY -15 to DAY + 14, groups LIM and LIM+IVM received 200 mg/kg of body weight/day of LIM via oral. On DAY 0, a single dose of IVM at 200 µg/kg of body weight was administered orally to groups IVM and LIM+IVM. On DAY + 7 and DAY + 14, fecal egg counts (FEC) were performed and on DAY + 14 animals were euthanized for total worm count (TWC), worm length, fecundity of females, and Pgp-9 gene expression. On DAY + 7, group LIM+IVM had 96.29% efficacy based on Fecal Egg Count Reduction TEST (FECRT) and a highly significant reduction in FEC (P = 0.0005) when compared to CTL. On DAY + 14, the efficacy of LIM+IVM was 82.87% on FECRT, although no differences were found among groups for FEC, TWC, worm length, or Pgp-9 gene expression. Female worms from the CTL group had higher egg counts in their uterus when compared to LIM. No differences were found for hematological or biochemical parameters, body weight, or weight gain among groups. Thus, LIM given daily at 200 mg/kg was safe for animals and, when combined with IVM, decreased egg shedding and could reduce pasture contamination, although it was unable to kill multidrug-resistant H. contortus.

Evaluation of encapsulated anethole and carvone in lambs artificially- and naturally-infected with Haemonchus contortus.

Molecules from natural sources, such as essential oils, have shown activity against parasites in vitro, but have not yet been explored extensively in vivo. Anethole and carvone (10% each), encapsulated with 80% of a solid matrix, referred to as EO (encapsulated oils), were tested in vivo in 2 experiments. In Experiment 1: Lambs were artificially infected with multidrug resistant Haemonchus contortus, or left uninfected, and treated (or not) with 50 mg/kg bw (body weight) of EO in a controlled environment. Thirty-two male lambs were kept in individual cages for a period of 45 days, after which animals were evaluated for parasitological, hematological, toxicological, and nutritional parameters. After 45 days of treatment, EO at 50 mg/kg bw provided a significant (P ≤ 0.05) reduction in fecal egg count (FEC). Although FEC was reduced, animals from both treatments had similar counts of total adult worms. The low FEC was caused probably by a significant reduction (P ≤ 0.05) in both male worm size and female fecundity. Dry matter intake of uninfected controls was significantly (P ≤ 0.05) reduced, although no toxicity was observed in treated animals. Thus, in Experiment 2, conducted for five months we used an EO dose of 20 mg/kg bw. Thirty-four weaned lambs, free of parasites, were divided in two groups and kept in collective pens. One group received EO at 20 mg/kg bw mixed with concentrate for 5 months and the other was kept as a control group (CTL). Parasitological and hematological parameters as well as body weight were evaluated. In the first 2.5 months, CTL and EO groups were confined, and both presented similar clinical parameters. Then, animals were allotted to graze on contaminated pastures to acquire natural infection for the next 2.5 months. The infection was patent after 25 days and both groups had similar decreases in weight gain, increases in FEC, and decreases in blood parameters. Coprocultures from CTL and EO groups established that parasite population was 90% Haemonchus sp. We concluded that the technology of encapsulation is safe and practical to deliver to lambs at the farm level and anethole and carvone at 50 mg/kg bw caused a significant decrease in FEC and, consequently, in pasture contamination by free living stages of H. contortus. However, EO at 20 mg/kg bw was not effective to prevent or treat sheep naturally-infected with gastrointestinal nematodes.

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)

Terminalia catappa: Chemical composition, in vitro and in vivo effects on Haemonchus contortus.

Haemonchus contortus is the most important nematode in small ruminant systems, and has developed tolerance to all commercial anthelmintics in several countries. In vitro (egg hatch assay) and in vivo tests were performed with a multidrug strain of Haemonchus contortus using Terminalia catappa leaf, fruit pulp, and seed extracts (in vitro), or pulp and seed powder in lambs experimentally infected with H. contortus. Crude extracts from leaves, fruit pulp and seeds obtained with 70% acetone were lyophilized until used. In vitro, the extracts had LC50=2.48µg/mL (seeds), LC50=4.62µg/mL (pulp), and LC50=20µg/mL (leaves). In vitro, seed and pulp extracts had LC50 similar to Thiabendazole (LC50=1.31µg/mL). Condensed tannins were more concentrated in pulp extract (183.92g of leucocyanidin/kg dry matter) than in either leaf (4.6g) or seed (35.13g) extracts. Phytochemical tests established that all extracts contained alkaloids, flavonoids, saponins, phenols, and terpenoids. Based on these results, in vivo tests were performed to evaluate the anthelmintic activity of T. catappa whole fruit (pulp+seed) powder. Male Santa Ines lambs were artificially infected with multidrug-resistant H. contortus and divided, according to similar fecal egg count (FEC) and weight, into two groups: Control (infected/untreated) and treated (infected/treated with whole fruit powder). Whole fruit powder was mixed with concentrate and provided at 2g/kg of body weight (BW) for five days. After treatment, parasitological analysis (FEC and egg hatch assay), renal profile (urea and creatinine), liver profile (aspartate aminotransferase) and BW were determined. In vitro (based on LC50), seed/pulp extracts had ovicidal effect similar to Thiabendazole but whole fruit powder had no anthelmintic effect on adult nematodes in the abomasum. We discuss the plausible causes of the lack of in vivo activity.

Synergistic interaction of ten essential oils against Haemonchus contortus in vitro.

Anthelmintic resistance in sheep gastrointestinal nematodes is a worldwide problem. Multi-drug resistant haemonchosis is the most serious impediment for small ruminant systems, and there are no new drug candidates currently under development. Molecules from natural sources have demonstrated anthelmintic activity against parasites. In this work, the monoterpenoids carvacrol, carvone, cineole, linalool, limonene, and thymol and the phenylpropanoids cinnamaldehyde, anethole, vanillin, and eugenol were assessed individually or in mixtures of ten binary, three ternary, and three quaternary combinations using the in vitro egg hatch assay with eggs of a multi-drug resistant strain of Haemonchus contortus. The main objective of this study was to identify the most effective interaction among essential oils with the greatest individual anthelmintic efficacy and to determine the most powerful combinations. The essential oils were ranked by their 50% lethal concentration (LC50) as follows (mg/mL): cinamaldehyde (0.018), anethole (0.070), carvone (0.085), carvacrol (0.11), thymol (0.13), linalool (0.29), vanillin (0.57), eugenol (0.57), cineole (4.74), and limonene (207.5). Quantification of synergism, additive effect, and antagonism were calculated for binary, ternary, and quaternary combinations. The best anthelmintic effect resulting from synergistic activity among 16 different combinations was for cinnamaldehyde:carvacrol (CL50 0.012mg/mL) and anethole:carvone (CL50 0.013mg/mL). These results indicate that these binary combinations would be promising to be tested in sheep infected with H. contortus.

Recommendations for Vaccination in Adult Patients with Systemic Inflammatory Rheumatic Diseases from the Portuguese Society of Rheumatology.

BACKGROUND: Serious infections are a major cause of morbidity and mortality in systemic inflammatory rheumatic disease (SIRD) patients. Although vaccination may prevent numerous infections, vaccination uptake rates are low in this group of patients. OBJECTIVES: To develop evidence-based recommendations for vaccination in SIRD patients. METHODS: We searched MEDLINE (until 31 October 2014) and EMBASE (until 14 December 2014) databases, as well as the ACR and EULAR congress abstracts (2011-2014). Patients with any systemic inflammatory rheumatic disease were included and all vaccines were considered. Any safety and efficacy outcomes were admitted. Search results were submitted to title and abstract selection, followed by detailed review of suitable studies. Data were subsequently pooled according to the type of vaccine and the SIRD considered. Results were presented and discussed by a multidisciplinary panel and systematic literature review (SLR)-derived recommendations were voted according to the Delphi method. The level of agreement among rheumatologists was assessed using an online survey. RESULTS: Eight general and seven vaccine-specific recommendations were formulated. Briefly, immunization status should routinely be assessed in all SIRD patients. The National Vaccination Program should be followed and some additional vaccines are recommended. To maximize the efficacy of vaccination, vaccines should preferably be administered 4 weeks before starting immunosuppression or, if possible when disease activity is controlled. Non-live vaccines are safe in SIRD, including immunosuppressed patients. The safety of live attenuated vaccines in immunosuppressed patients deserves further ascertainment, but might be considered in particular situations. DISCUSSION: The present recommendations combine scientific evidence with the multidisciplinary expertise of our taskforce panel and attained desirable agreement among Portuguese rheumatologists. Vaccination recommendations need to be updated on a regular basis, as more scientific data regarding vaccination efficacy and safety, emergent infectious threats, new vaccines as well as new immunomodulatory therapies become available.

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.

Rumen fermentation and production effects of Origanum vulgare L. leaves in lactating dairy cows.

A lactating cow trial was conducted to study the effects of dietary addition of oregano leaf material (Origanum vulgare L.; OV; 0, control vs. 500 g/d) on ruminal fermentation, methane production, total tract digestibility, manure gas emissions, N metabolism, organoleptic characteristics of milk, and dairy cow performance. Eight primiparous and multiparous Holstein cows (6 of which were ruminally cannulated) were used in a crossover design trial with two 21-d periods. Cows were fed once daily. The OV material was top-dressed and mixed with a portion of the total mixed ration. Cows averaged 80 ± 12.5 d in milk at the beginning of the trial. Rumen pH, concentration of total and individual volatile fatty acids, microbial protein outflow, and microbial profiles were not affected by treatment. Ruminal ammonia-N concentration was increased by OV compared with the control (5.3 vs. 4.3mM). Rumen methane production, which was measured only within 8h after feeding, was decreased by OV. Intake of dry matter (average of 26.6 ± 0.83 kg/d) and apparent total tract digestibly of nutrients did not differ between treatments. Average milk yield, milk protein, lactose, and milk urea nitrogen concentrations were unaffected by treatment. Milk fat content was increased and 3.5% fat-corrected milk yield tended to be increased by OV, compared with the control (3.29 vs. 3.12% and 42.4 vs. 41.0 kg/d, respectively). Fat-corrected (3.5%) milk feed efficiency and milk net energy for lactation (NE(L)) efficiency (milk NE(L) ÷ NE(L) intake) were increased by OV compared with the control (1.64 vs. 1.54 kg/kg and 68.0 vs. 64.4%, respectively). Milk sensory parameters were not affected by treatment. Urinary and fecal N losses, and manure ammonia and methane emissions were unaffected by treatment. Under the current experimental conditions, supplementation of dairy cow diets with 500 g/d of OV increased milk fat concentration, feed and milk NE(L) efficiencies, and tended to increase 3.5% fat-corrected milk yield. The sizable decrease in rumen methane production with the OV supplementation occurred within 8h after feeding and has to be interpreted with caution due to the large within- and between-animal variability in methane emission estimates. The OV was introduced into the rumen as a pulse dose at the time of feeding, thus most likely having larger effect on methane production during the period when methane data were collected. It is unlikely that methane production will be affected to the same extent throughout the entire feeding cycle.

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.

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.

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.