Ophionereis reticulata (Echinodermata, Ophiuroidea)

This study compared the arm regeneration frequencies in two different populations of Ophionereis reticulata (Say, 1825) in São Sebastião, Southeast Brazil and observed arm regeneration between age classes (juvenile and adults) and sexes (male and female). From the 1,170 individuals sampled, 1,089 (92.2%) showed signs of arm regeneration. The relative frequencies of regenerating arms in the two areas were not different (Baleeiro Isthmus: 91.3% and Grande Beach: 99.5%). Both areas also presented similar values for the number of arms regenerating/individual and in the frequency of regenerating individuals. The major part of the regenerating scars was concentrated in the distal portion of the arm. Sub-lethal predation is most likely the cause to the high rates of arm regeneration in O. reticulata. There was no significant differences in the regeneration rates between females (3.57 ± 1.36 arms regenerating/individual) and males (3.47 ± 1.42).

Mendelian inheritance, linkage and linkage disequilibrium in microsatellite loci of Copaifera langsdorffii Desf.

Copaifera langsdorffii is a Neotropical tree with wide distribution in the Brazilian Atlantic rain forest and savanna. Although eight microsatellite loci (SSR) were developed in 2000 and have been widely used since then, there is yet no information about their inheritance, linkage and linkage disequilibrium (LD). Through the analysis of 28 open-pollinated (OP) progenies, the SSR loci revealed Mendelian inheritance and independent assortment. Using these progenies, young and adult trees LD was mainly detected in OP progenies. Our results show clear evidence that the eight SSR loci can be used without restriction in genetic diversity, mating system and parentage analysis.

Antimicrobial Activity of Terpenoids from Copaifera langsdorffii Desf. Against Cariogenic Bacteria

In the present work, the anticariogenic activities of nine labdane type-diterpenes and four sesquiterpenes were investigated. Among these metabolites, (-)-copalic acid (CA) was the most active compound displaying MIC values very promising (ranging from 2.0 to 6.0 mu g/mL) against the main microorganisms responsible for dental caries: Streptococcus salivarius, S. sobrinus, S. mutans, S. mitis, S. sanguinis and Lactobacillus casei. Time kill assays performed with CA against the primary causative agent (S. mutans) revealed that, in the first 12 h, this compound only inhibits the growth of the inoculum (bacteriostatic effect). However, its bactericidal effect is clearly noted thereafter (between 12 and 24 h). Also, CA did not show a synergistic effect when combined with the anticariogenic gold standard (chlorhexidine, CHD) in the checkerboard assays against S. mutans. In conclusion, the results points out CA as an important metabolite in the search for new effective anticariogenic agents. Copyright (C) 2010 John Wiley & Sons, Ltd.

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

The ecological and economic importance of oleoresin produced by Copaifera langsdorffii is well established. This study aims to investigate the ontogeny, anatomy and ultrastructure of the internal glands of C. langsdorffii during plant development. Samples were processed for light and electron microscopy and a specific technique was applied to impregnate endomembranes. Internal secretory glands were observed in the hypocotyl, epicotyl and eophylls of seedlings, and in the primary stem, pulvinus, petiole, rachis and leaf blade of adult plants. Canals and cavities show differential distribution. They arise from ground meristem cells, and the lumen is first formed by schizogenesis followed by later schizolysigenous development. The dense cytoplasm of epithelial cells shows mitochondria, plastids without thylakoids, polyribosomes and endoplasmic reticulum. A periplastidial reticulum was also observed. Secretion is released by eccrine, granulocrine and holocrine processes. Lipophilic and hydrophilic compounds were histochemically detected in both canals and cavities, whereas resin was detected only in canals. The presence of these substances has been associated with plants` defences against dehydration, as well as against attacks from herbivores and pathogens, from seedling stage onwards. (C) 2011 Elsevier GmbH. All rights reserved.

ELUCIDATING ENIGMATIC FLORAL ISSUES IN COPAIFERA LANGSDORFFII DESF. (LEGUMINOSAE, CAESALPINIOIDEAE)

This study focuses on the floral development of Copaifera langsdorffii to elucidate uncertain features in its floral morphology, such as the tetramerous calyx, lack of petals, blackened anthers and their supposed sterility, as well as polyembryony. Buds and flowers were dissected and prepared for examination under scanning electron and light microscopes. The floral apex initiates two bracteoles, five sepals, five petals, five outer stamens, five inner stamens, and one carpel. Order is helical for sepals, reversed unidirectional for the petals, and unidirectional for two whorls of stamens. The tetramerous calyx results from the union of two adaxial sepal primordia, which forms one large sepal and three other smaller sepals. Although the flower lacks petals, the petal primordia are initiated but do not elongate like the other floral organs, remaining as petal rudiments. Ten stamens are formed in two distinct whorls. Formation within each whorl is almost simultaneous, and the inner whorl is formed shortly after the outer. During organ elongation, the inner stamen primordia bases are reoriented outward, resulting in a single whorl of stamens. The darkened anthers have viable pollen grains. Thus, there is no relation between sterility and the dark coloration of the anthers. No signs of extranumerary embryos are observed; therefore, polyembryony is not confirmed. Although studies on floral development of Detarieae have been reported, few Neotropical genera of the tribe (such as Copaifera) have been studied.

Laboratory evaluation of the physicochemical properties of a new root canal sealer based on Copaifera multijuga oil-resin

Aim To compare a new root canal sealer based on Copaifera multijuga oil-resin (Biosealer) using three other established sealers (Sealer 26 (R), Endofill (R) and AH plus (R)) in terms of their physicochemical properties. Methodology The study was carried out according to the requirements of Specification Number 57 of the American Dental Association (ADA) and consisted of the following tests: setting time, flow, film thickness, dimensional stability, radiopacity and solubility/disintegration. Data were analysed statistically using anova and Tukey`s test for multiple comparisons. The significance level was set at 5% for all analyses. Results Sealer 26 (R) and AH Plus (R) had the longest setting time (P < 0.05). All materials presented flow in with the ADA`s guidelines. Regarding film thick-ness, Sealer 26 (R) did not have a satisfactory performance, as it had a higher mean value than the maximum allowed by the ADA (0.05 mm), being significantly different from the other materials (P < 0.05), which had mean values for film thickness in accordance with the ADA`s recommendations. Regarding the solubility and disintegration, only Endofill (R) did not meet the ADA`s specifications and presented the worst results of all materials (P < 0.05). Sealer 26 (R) presented the greatest dimensional changes and differed significantly from all other sealers (P < 0.05). Biosealer had the lowest radiopacity values and was significantly different from the other sealers (P < 0.05). Conclusion The experimental sealer based on Copaifera multijuga oil-resin presented satisfactory results in the physicochemical tests required by the ADA.

Reproductive cycle of Ophionereis reticulata (Ophiuroidea, Echinodermata) on the southeast coast of Brazil

The reproductive cycle of Ophionereis reticulata, a common sediment-rocky shore-interface ophiuroid, was examined monthly from January 2002 to January 2003 at Praia Grande beach (Sao Sesbatiao, state of Sao Paulo, Brazil). Mature individuals were found from January 2002 to April 2002 and from November 2002 to January 2003. Spawning was regcorded from January 2002 to March 2002 and from November 2002 to January 2003. Mature sperm is still present in April which could be involved in the fertilization of the last oocytes spawned in March and April. November and December marked the final point in the maturation process, with a high concentration of yolk and lipid nutrients in the oocytes. Ophionereis reticulata showed, for the period analyzed, a single spawning period, during spring and summer.

Antineoplasic activity of Copaifera multijuga oil and fractions against ascitic and solid Ehrlich tumor

The aim of this study was to investigate the effect of chronic treatment with C. multijuga oil on Ehrlich tumor evolution. C multijuga was fractionated in a KOH impregnated silica gel column chromatography to give three distinct fractions, i.e., hexanic, chloroformic, and methanolic, mainly composed by hydrocarbon sesquiterpenes, oxygenated sesquiterpenes and acidic diterpenes, respectively. Results demonstrated that the C multijuga oil, the hexanic, and chloroformic fractions did not develop toxic effects. The oil, hexanic and chloroformic fractions (doses varying between 100 and 200 mg/kg) showed antineoplasic properties against Ehrlich ascitic tumor (EAT) and solid tumor during 10 consecutive days of treatment inhibiting ascitic tumor cell number, reverting medulla and blood cell counts to values similar to control group, and inhibiting the increase on several inflammatory mediators (total protein, PGE(2), nitric oxide, and TNF) on ascitic fluid. The treatment also inhibited the increase in paw volume on tumor-inoculated mice. In conclusion, C. multijugo as well as its fractions demonstrated antineoplasic effect even after oral administration confirming its use by traditional medicine. (C) 2008 Elsevier Ireland Ltd. All rights reserved.

Isolation, chemical composition and anti-inflammatory activity of Copaifera langsdorffii desf. fruit peels essential oil according to successive hydrodistillations

In this study, the isolation and characterization of chemical composition of C. langsdorffii (copaiba) fruit peel volatile oil from Alfenas - MG was carried out according to successive hydrodistillations, likewise the anti-inflammatory activity was evaluated by rat paw edema model. The chemical composition of the oil was determined by GC-MS analysis. The major components were (E)-caryophyllene, germacrene B, 1,5-epoxysalvial-4(14)-ene, caryophyllene oxide and isospathulenol. The inflammatory process was inhibited in 39 +/- 6 % (p<0.05, Tukey-Kramer) by one sample of essential oil. The C. langsdorffii fruit peel is a rich source of essential oil (3.8% w/w), and due to potential industrial interest of its components this ecotype presents feasible crop seeking productivity and it could constitutes in a good alternative to the sustainable use of this specie.

Tree performance and fruit yield and quality of `Okitsu` Satsuma mandarin grafted on 12 rootstocks

The citriculture in Brazil, as well as in other important regions in the world, is based on very few mandarin cultivars. This fact leads to a short harvest period and higher prices for off-season fruit. The `Okitsu` Satsuma (Citrus unshiu Marc.) is among the earliest ripening mandarin cultivars and it is considered to be tolerant to, citrus canker (Xanthomonas citri subsp. citri Schaad et al.) and to citrus variegated chlorosis (Xylella fastidiosa Wells et al.). Despite having regular fruit quality under hot climate conditions, the early fruit maturation and absence of seeds of `Okitsu` fruits are well suited for the local market in the summer(December through March), when the availability of citrus fruits for fresh consumption is limited. Yet, only a few studies have been conducted in Brazil on rootstocks for `Okitsu`. Consequently, a field trial was carried out in Bebeclouro, Sao Paulo State, to evaluate the horticultural performance of `Okitsu` Satsuma mandarin budded onto 12 rootstocks: the citrandarin `Changsha` mandarin (Citrus reticulata Blanco) x Poncirus trifoliata `English Small`: the hybrid Rangpur lime (Citrus limonia Osbeck) x `Swingle` citrumelo (P. trifoliata (L.) Raf. x Citrus paradisi Macfad.); the trifoliates (P. trifoliata (L) Raf)`Rubidoux`,`FCAV` and `Flying Dragon`(P. trifoliata var. monstrosa); the mandarins `Sun Chu Sha Kat`(C. reticulata Blanco) and `Sunki`(Citrus sunki (Hayata) Hort. ex. Tanaka); the Rangpur limes (C. limonia Osbeck) `Cravo Limeira` and `Cravo FCAV`;`Carrizo` citrange (Citrus sinensis x P. trifoliata), `Swingle` citrumelo (P. trifoliata x C. paradisi), and `Orlando` tangelo (C. paradisi x Citrus tangerina cv. `Dancy`). The experimental grove was planted in 2001, using a 6 m x 3 m spacing, in a randomized block design. No supplementary irrigation was applied. Fruit yield, canopy volume, and fruit quality were assessed for each rootstock. A cluster multivariate analysis identified three different rootstock pairs with similar effects on plant growth, yield and fruit quality of `Okitsu` mandarin. The `Flying Dragon `trifoliate had a unique effect over the `Okitsu` trees performance, inducing lower canopy volume and higher yield efficiency and fruit quality, and might be suitable for high-density plantings. The `Cravo Limeira` and `Cravo FCAV` Rangpur limes induced early-ripening of fruits, with low fruit quality. `Sun Chu Sha Kat` and `Sunki` mandarins and the `Orlando` tangelo conferred lower yield efficiency and less content of soluble solids for the latter rootstock. (C) 2009 Elsevier B.V. All rights reserved.

Horticultural performance of `Folha Murcha` sweet orange onto twelve rootstocks

Despite its outstanding position, the Brazilian citriculture is established on a very limited pool of varieties that limits its expansion and restricts the fruit availability throughout the year. This situation determines the urgent necessity of developing alternative scion and rootstock cultivars, with good performance under local conditions. `Folha Murcha` sweet orange (Citrus sinensis (L.) Osbeck) is a late-harvest cultivar, suitable both for the juice processing industry and the fresh fruit market, being described as tolerant to citrus canker (Xanthomonas citri subsp. citri Schaad et al.), and less affected by citrus variegated chlorosis (Xylella fastidiosa Wells et al.). A study was conducted in Bebedouro, Sao Paulo State, Brazil, to evaluate the horticultural performance of `Folha Murcha` sweet orange budded onto 12 rootstocks: the citrandarin `Changsha` mandarin (Citrus reticulata Blanco) x Poncirus trifoliata `English Small`: the hybrid `Rangpur` lime (Citrus limonia Osbeck) x `Swingle` citrumelo (P. trifoliata (L.) Raf x Citrus paradisi Macfad.); the trifoliates (P. trifoliata (L.) Raf.)`Rubidoux`, `FCAV`, and `Flying Dragon` (P. trifoliata var. monstrosa); the `Sun Chu Sha Kat` mandarin (C. reticulata Blanco); the `Sunki` mandarin (Citrus sunki (Hayata) Hart. ex. Tanaka); the `Rangpur` limes (C. limonia Osbeck) `Cravo Limeira` and `Cravo FCAV`; `Carrizo` citrange (C. sinensis x P. trifoliata), `Swingle` citrumelo (P. trifoliata x C. paradisi), and `Orlando` tangelo (C. paradisi x Citrus tangerina cv. `Dancy`). The experimental grove was planted in 2001, using a 7 m x 4 m spacing, in a randomized block design, with five replications and two plants per plot. No supplementary irrigation was applied. Fruit yield, canopy volume, tree tolerance to drought and to citrus variegated chlorosis, and fruit quality were assessed for each rootstock. Trees grafted onto the `Flying Dragon` trifoliate were smaller in size, but had largest yield efficiency when compared to those grafted onto other rootstocks. Lower alternate bearing index was observed on trees budded onto `Cravo FCAV` `Rangpur` lime. Both `Rangpur` lime rootstocks and the `Sunki` mandarin induced higher tree tolerance to drought. The `Flying Dragon` trifoliate induced better fruit quality and higher tolerance to citrus variegated chlorosis (CVC) to `Folha Murcha` trees. A cluster multivariate analysis identified three groups of rootstocks with similar effects on `Folha Murcha` tree performance. Among the 12 evaluated rootstocks, the `Flying Dragon` trifoliate has a unique effect on plant growth, tolerance to drought and CVC, fruit yield and fruit quality of `Folha Murcha` trees, and may be better suited for high-density plantings. (C) 2011 Elsevier B.V. All rights reserved.

Endophytic population of Pantoea agglomerans in citrus plants and development of a cloning vector for endophytes

Harmless bacteria inhabiting inner plant tissues are termed endophytes. Population fluctuations in the endophytic bacterium Pantoea agglomerans associated with two species of field cultured citrus plants were monitored over a two-year period. The results demonstrated that populations of P. agglomerans fluctuated in Citrus reticulata but not C. sinensis. A cryptic plasmid pPA3.0 (2.9 kb) was identified in 35 out of 44 endophytic isolates of P. agglomerans and was subsequently sequenced. The origins of replication were identified and nine out of 18 open reading frames (ORFs) revealed homology with described proteins. Notably, two ORFs were related to cellular transport systems and plasmid maintenance. Plasmid pPA3.0 was cloned and the gfp gene inserted to generate the pPAGFP vector. The vector was introduced into P. agglomerans isolates and revealed stability was dependent on the isolate genotype, ninety-percent stability values were reached after 60 hours of bacterial cultivation in most evaluated isolates. In order to definitively establish P. agglomerans as an endophyte, the non-transformed bacterium was reintroduced into in vitro cultivated seedlings and the density of inner tissue colonization in inoculated plants was estimated by bacterium re-isolation, while the tissue niches preferred by the bacterium were investigated by scanning electronic microscopy (SEM). Cells from P. agglomerans (strain ARB18) at similar densities were re-isolated from roots, stems and leaves and colonization of parenchyma and xylem tissues were observed. Data suggested that P. agglomerans is a ubiquitous citrus endophyte harboring cryptic plasmids. These characteristics suggest the potential to use the bacterium as a vehicle to introduce new genes in host plants via endophytic bacterial transformation.

Validation of a gas chromatographic method to quantify sesquiterpenes in copaiba oils

Copaifera species (Leguminoseae) are popularly known as ""copaiba"" or ""copaiva"". The oleoresins obtained from the trunk of these species have been extensively used in folk medicine and are commercialized in Brazil as crude oil and in several pharmaceutical and cosmetic products. This work reports a complete validated method for the quantification of beta-caryophyllene, alpha-copaene, and alpha-humulene in distinct copaiba oleoresins available commercially. Thus, essential oil samples (100 mu L) were dissolved in 20 mL of hexanes containing internal standard (1,2,4,5-tetramethylbenzene, 3.0 mM) in a 25 mL glass flask. A 1 mu L aliquot was injected into the GC-FID system. A fused-silica capillary column HP-5, coated with 5% phenylmethylsiloxane was used for this study. The developed method gave a good detection response with linearity in the range of 0.10-18.74 mM. Limits of detection and quantitation variety ranged between 0.003 and 0.091 mM. beta-Caryophyllene, alpha-copaene, and alpha-humulene were recovered in a range from 74.71% to 88.31%, displaying RSD lower than 10% and relative errors between -11.69% and -25.30%. Therefore, this method could be considered as an analytical tool for the quality control of different Copaifera oil samples and its products in both cosmetic and pharmaceutical companies. (C) 2010 Elsevier B.V. All rights reserved.

Taxonomic revision of Gripopteryx (Pictet, 1841) (Plecoptera: Gripopterygidae)

The genus Gripopteryx is revised and two new species are described, G. japi n. sp. from southeastern Brazil, and G. clemira n. sp. from northeastern Brazil. The male of G. brasiliensis Samal, 1921 and the female of G. flinti Froehlich, 1993 are described. New figures for G. maculosa Jewett, 1960, for the male of G. reticulata Brauer, 1866, and for the female of G. brasiliensis are provided. Keys to adult males and to nymphs are provided.

Role of olfaction and vision cues in feeding behavior and alarm reaction in the catfish pintado, Pseudoplatystoma corruscans

Experiments were performed to investigate senses that are essential for mediating fright reaction and food behavior in Pseudoplatystoma corruscans, pintado. The dilemma ""to feed or to flee"" was also analyzed in fishes with intact and sectioned olfactory tracts, stimulated by alarm substance extracts and food. Fishes were arranged into five groups: fish with intact lateral olfactory tracts (LOT), fish with intact medial olfactory tract (MOT), fish with tracts totally sectioned (TOTAL, both LOT and MOT), sham operated, and nonoperated fish. The five groups were submitted to either alarm substance extract and food stimulus or to distilled water (control) and food stimulus. Fish reacted to food independently of which tract (LOT, MOT or TOTAL) was sectioned; vision seems necessary and elemental to detect and deflagrate food response. Latency of the responses to each reaction was different between groups. None of the fish with sectioned tracts reacted to alarm substance extract, while sham- and nonoperated fish showed the typical alarm behavior response, leading to the conclusion that olfaction is essential for mediating alarm response. These results indicate that others sense systems (e.g., vision) are sufficient to trigger and elicit feeding behavior and that olfaction is not necessary to fully maintain food detection to qualitative and quantitative extent. However, olfactory tract integrity seems to be required for mediation of alarm reaction in P. corruscans.