Detection and characterization of fibropapilloma associated herpesvirus of marine turtles in Rio Grande do Sul, Brazil

Fibropapillomatosis (FP) is a benign tumoral disease that affects sea turtles, hampering movement, sight and feeding, ultimately leading to death. In Brazil, the disease was described for the first time in 1986. Research suggests the involvement of a herpesvirus in association with environmental and genetic factors as causal agents of FP. The objective of the present study was to detect and characterize this herpesvirus in sea turtles living in the coast of state Rio Grande do Sul (RS), Brazil. From October 2008 to July 2010, 14 turtles were observed between the beaches of Torres and Tavares, of which 11 were green turtles (Chelonia mydas) and 3 were loggerhead turtles (Caretta caretta). All turtles were young and mean curved carapace length was 37.71±7.82cm, and varied from 31 to 55cm. Only one green turtle presented a 1cm, papillary, pigmented fibropapilloma. Skin and fibropapilloma samples were analyzed by conventional and real time PCR assays to detect and quantify herpesvirus. All skin samples were negative, though the fibropapilloma specimen was positive in both tests. Viral load was 9,917.04 copies of viral genome per milligram of tissue. The DNA fragment amplified from the fibropapilloma sample was sequenced and allocated in the Atlantic phylogeographic group. This study reports the first molecular characterization of herpesvirus associated with fibropapilloma in turtles from the coast of RS.

Reserve substances and storage of Cyathea delgadii Sternb. spores

The spores used were collected from a population of trees of Cyathea delgadii Sternb. growing in a gallery forest at the Reserva Biológica e Estação Experimental de Moji Guaçu, São Paulo state, Brazil (22°18’S and 47°11’W). The germination of spores of Cyathea delgadii decreases with time when kept in closed bottles under storage at 4°C in darkness. Germination is still very high after storage for one year. Spores stored for three years do not germinate. The results also show a decrease in soluble proteins and an increase in starch after several months storage.

Purification and characterisation of a lectin from the red marine alga Pterocladiella capillacea (S.G. Gmel.) Santel. & Hommers.

A lectin present in the marine red alga Pterocladiella capillacea was purified and characterised by extraction of soluble proteins (crude extract) in 20 mM Tris-HCl buffer, pH 7.5. Among the analysed erythrocytes (human blood group A, B and O and the animals ox, goat, chicken and rabbit) the lectin agglutinated specifically rabbit erythrocytes. The hemagglutinating activity assay showed that the lectin was not dependent on divalent cations and was shown to be inhibited by the glycoproteins avidin and mucin. The purification procedure was conduced by precipitation of the crude extract with 80% saturation ammonium sulfate (F0/80) followed by affinity chromatography on guar-gum column. The lectin of P. capillacea was purified 14.5 fold and had a recovery of 27.4% of the original total specific activity present in the crude extract. The absence of carbohydrate suggested that the lectin is not a glycoprotein. The molecular mass of P. capillacea lectin, determined by gel filtration, was 5.8 kDa. SDS-PAGE in the presence of ß-mercaptoethanol gave one band, indicating that the native lectin is a monomeric protein. The activation energy of denaturation process (D G') was calculated to be 106.87 kJ . mol-1 at 70 ºC.

Lecythidaceae Poit. in the Tupé Sustainable Development Reserve, Manaus, Brazil

Lecythidaceae is the family of the Brazil nut, and comprises about 300 species belonging to 17 genera with pantropical distributions. One hundred and twenty-two species belonging to nine genera are distributed throughout Brazil, demonstrating its greatest diversity in the Amazon rainforest where Lecythidaceae is also one of the most abundant families. It is usually difficult to collect fertile material from these trees because of their canopy heights, and species determinations using sterile material can be complex because of their morphological similarities. There have been relatively few studies of this family even though it is one of the most important groups in the Amazon region, and a detailed taxonomic treatment of the species of Lecythidaceae in the Tupé Sustainable Development Reserve was therefore the goal of the present work. Ten species were found, Allantoma lineata (Mart. ex O.Berg) Miers, Bertholletia excelsa Bonpl., Couratari tenuicarpa A.C.Sm., Lecythis poiteaui O. Berg; and six species of Eschweilera, the richest genus. The descriptions and identification keys of the species used 56 characters. The main reproductive characters useful for distinguishing the species were the pubescence of the inflorescence rachis, pedicel length and trichomes presence, floral symmetry, hood type, filament shape, stigma shape, fruit shape and size, and aril type. The most diagnostic vegetative characters were the type and color of the outer bark, inner bark color, midrib prominence, and petiole shape and pubescence.

Isolation and characterization of a reserve protein from the seeds of Opuntia ficus-indica (Cactaceae)

We describe here the isolation and characterization of a major albumin from the seeds of Opuntia ficus-indica (Cactaceae). This protein has a molecular mass of 6.5 kDa and was isolated by a combination of gel filtration chromatography and reverse-phase HPLC. The amino acid composition of this protein was determined and it was shown to have similarities with the amino acid composition of several proteins from the 2S albumin storage protein family. The N-terminal amino acid sequence of this protein is Asp-Pro-Tyr-Trp-Glu-Gln-Arg.

Antinociceptive properties in mice of a lectin isolated from the marine alga Amansia multifida Lamouroux

The antinociceptive effects of a lectin (LEC) isolated from the marine alga Amansia multifida were determined in Swiss mice. The LEC (1, 5, and 10 mg/kg) inhibited acetic acid-induced abdominal writhings in a dose-dependent manner after intraperitoneal or oral administration. A partial but significant inhibition of writhings was observed after the combination of LEC (10 mg/kg) with avidin (1 mg/kg), a potent inhibitor of the hemmaglutinant activity of the lectin. However, total writhing inhibition was demonstrable in the group of mice treated with LEC plus mannose (1 mg/kg), as compared to LEC alone or to control groups. Furthermore, avidin and mainly mannose also play a role in antinociception, somehow facilitating the interaction of LEC with its active cell sites. In the formalin test, although both phases of the response were significantly inhibited, the effect of LEC was predominant during phase 2, causing inhibition of licking time that ranged from 48 to 88% after oral (5 and 10 mg/kg) and intraperitoneal (1 to 5 mg/kg) administration. As is the case with morphine, the effect of LEC (2 mg/kg) was reversed by naloxone (2 mg/kg), indicating the involvement of the opioid system. LEC was also effective in the hot-plate test, producing inhibitory responses to the thermal stimulus, and its effects were blocked by naloxone. In the pentobarbital-induced sleeping time, although LEC did not alter the onset of sleep significantly, it increased the time of sleep within the same dose range compared to control. These results show that LEC presents antinociceptive effects of both central and peripheral origin, possibly involving the participation of the opioid system.

Effect of hormone replacement on exercise cardiopulmonary reserve and recovery performance in subclinical hypothyroidism

Subclinical hypothyroidism (SH) patients present cardiopulmonary, vascular and muscle dysfunction, but there is no consensus about the benefits of levothyroxine (L-T4) intervention on cardiopulmonary performance during exercise. The aim of the present study was to investigate the effects of L-T4 on cardiopulmonary exercise reserve and recovery in SH patients. Twenty-three SH women, 44 (40-50) years old, were submitted to two ergospirometry tests, with an interval of 6 months of normalization of thyroid-stimulating hormone (TSH) levels (L-T4 replacement group) or simple observation (TSH = 6.90 μIU/mL; L-T4 = 1.02 ng/dL). Patients with TSH >10 μIU/mL were excluded from the study to assure that they would receive treatment in this later stage of SH. Twenty 30- to 57-year-old women with no thyroid dysfunction (TSH = 1.38 μIU/mL; L-T4 = 1.18 ng/dL) were also evaluated. At baseline, lower values of gas exchange ratio reserve (0.24 vs 0.30; P < 0.05) were found for SH patients. The treated group presented greater variation than the untreated group for pulmonary ventilation reserve (20.45 to 21.60 L/min; median variation = 5.2 vs 25.09 to 22.45 L/min; median variation = -4.75, respectively) and for gas exchange ratio reserve (0.19 to 0.27; median variation = 0.06 vs 0.28 to 0.18; median variation = -0.08, respectively). There were no relevant differences in cardiopulmonary recovery for either group at baseline or after follow-up. In the sample studied, L-T4 replacement improved exercise cardiopulmonary reserve, but no modification was found in recovery performance after exercise during this period of analysis.

Effects of filtration methods on the neutralization yield of crude marine fish oil

The evaluation of filtration methods on the yield and oleochemicals characteristics on the crude fish oil from the soapstock of marine fish for purposes of nutrition has been conducted in this work. The analytical properties of the crude and the neutralized oil with three excesses of sodium hydroxide (NaOH, 20%, 40% and 60%) were carried out on two different ways with organza and the “glass wool”. The neutralization of the oil brought about a notable improvement in the analytical properties of the oil. Thus, it leads to a high quality fish oil in terms of taste, colour, odours, shelf life and market value. Based on the improved characteristics of the oil, it could be suitable for applications in pharmaceutical and food industries.

Reserve mobilization during imbibition of stored Gliricidia sepium (Jacq.) Steud. (Leguminosae - Papilionoideae) seeds

Gliricidia sepium is a drought-tolerant species, easily multiplied by seeds, and has been exploited by farmers as a source of forage in the semi-arid region of northeast Brazil. The objective of the present study was to evaluate the effect of seed storage on the mobilization of reserves during imbibition of "Gliricidia" seeds. Freshly-harvested seeds were packed in kraft paper bags and stored for three and six months in the laboratory under ambient conditions (25 º C ± 3 T and 75% ± 3 RH). Cotyledons were isolated from imbibed seeds and macerated for the extraction and quantification of total soluble sugars, reducing sugars, sucrose and starch, as well as of proteins, amino acids and for amylase activity. Storage under these conditions resulted in an increase in seed water content although germination remained at relatively high levels (86%). Seed macromolecule levels showed significant variation with the storage period and imbibition and these variations were associated with a loss in seed viability due to inadequate storage conditions.