Impacts of the insecticide thiamethoxam on the native stingless bee Plebeia catamarcensis (Hymenoptera, Apidae, Meliponini).

Agricultural production and the indiscriminate use of insecticides such as thiamethoxam have put at risk the biodiversity and ecosystem services provided by bees, including native stingless species. Since most of the native species do not present economic importance, they may suffer "silent extinction", due to lack of monitoring of their colonies. Therefore, this study aimed to determine the lethal and sublethal concentrations of the insecticide thiamethoxam, with evaluation of its sublethal effects on mobility, in the stingless bee Plebeia catamarcensis (Holmberg, 1903). Foraging bees were collected and exposed to thiamethoxam to determine lethal (LC50) and sublethal concentrations. The 24 h LC50 was 0.408 ng a.i./µL, a value demonstrating that this species may be as sensitive as other stingless bees already studied. Sublethal concentrations influenced the locomotion abilities of the bees, making them hyperactive when exposed to LC50/10 and lethargic when exposed to LC50/100. The effects of sublethal concentrations on individuals may have collective consequences, especially in colonies with few individuals, as is the case of P. catamarcensis. The findings reinforce the hypothesis that thiamethoxam may contribute to the decline of native stingless bees, which can be significantly impacted when chronically exposed to agricultural production systems that use this insecticide, consequently affecting the ecosystem services provided by these bees.

In vitro antiproliferative activity of alkaloids isolated from Tabernaemontana catharinensis A.DC (Apocynaceae).

In this work, four alkaloids from the stem bark of T. catharinensis were isolated, namely: voacangine (1); ethyl apovincaminate (2); affinisine (3) and voachalotine (4). The alkaloids were tested in vitro for antiproliferative capacity in eight tumor cell lines: U251 (glioma), MCF-7 (breast), NCI-ADR/RES (drug resistant ovary), 786-0 (kidney), NCI-H460 (lung), HT-29 (colon), K562 (leukemia) and PC-3 (prostate) and a non-tumor keratinocyte cell line (HaCat). Antiproliferative activity was observed after 48 hours and results expressed as the concentration needed to induce 50% growth inhibition (GI50) in µM. The chemotherapy drug Doxorubicin was used as a standard. The alkaloid affinisine (3) was the most promising, showing moderate inhibition rates in addition to the cytotoxic and cytocidal effect against all strains tested. It also proved to be a very promising compound, showing high selectivity rates when compared to the non-tumor keratinocyte cell line (HaCat).

Constituents of leaves from Bauhinia curvula Benth. exert gastroprotective activity in rodents: role of quercitrin and kaempferol.

The Bauhinia genus is known as "Pata-de-Vaca" and a wide variety of these species are used in Brazilian folk medicine due to their gastroprotective properties. This study aimed to investigate the antiulcer efficacy of the hydroalcoholic extract from B. curvula (HEBC) leaves, as well as its semi-purified fraction (SPFr) and the contribution of their phytochemicals constituents for this effect. For that, ethanol 60%/HCl 0.3 M- and indomethacin-induced gastric ulcer were performed in rodents. Gastric ulcerated tissues were processed for histological, histochemical and biochemical analysis. The oral treatment with HEBC and SPFr decreased the gastric ulcer induced by ethanol/HCl in mice and by indomethacin (only HEBC) in rats. The gastroprotective effect of HEBC was abolished in mice pretreated with Nω-Nitro-L-arginine methyl ester, N-Ethylmaleimide, glibenclamide or indomethacin. Both HEBC and SPFr reduced myeloperoxidase activity in parallel with a decrease of lipoperoxides content at the site of the lesion. On the other hand, HEBC did not alter volume, pH, total acidity or pepsin activity of acid gastric secretion in rats, and neither inhibited the in vitro H(+),K(+)-ATPase activity. Additionally, the compounds identified and isolated from the SPFr, the flavonoids quercitrin (65%) and kaempferol (35%), were able to diminish the extent of ulcerated area induced by both ethanol/HCl and indomethacin. Taking together, these findings show that B. curvula extracts present gastroprotective effect, mainly explained by the presence of flavonoids quercitrin and kaempferol, which may possibly improve the defensive factors of gastric mucosa.

Croton argenteus preparation inhibits initial growth, mitochondrial respiration and increase the oxidative stress from Senna occidentalis seedlings.

Senna ocidentalis is a weed, native to Brazil, considered to infest crops and plantations, and is responsible for yield losses of several crops, particularly soybean. The aim of this work was to evaluate if the Croton argenteus extract and fractions possess phytotoxic activity on S. ocidentalis. The crude ethanolic extract (CEE) and its hexanic (HF), chloroformic (CLF) and ethyl acetate (EAF) fractions were tested in germination, growth, oxidative stress increase, Adenosine triphosphate, L-malate and succinate synthesis. The crude extract and its fractions slowed down the germination of S. ocidentalis and decreased the final percentage of germination. Oxidative stress was also increased in the seedlings, by an increase of catalase, peroxidase, superoxide dismutase, glutathione reductase and lipid peroxidation; and it became clear that the ethyl acetate fraction was more phytotoxic. The results indicate that the crude extract and fractions of C. argenteus compromise the mitochondrial energy metabolism, by the inhibition of mitochondrial ATP production, with a decrease in the production of L-malate and succinate. The ethyl acetate fraction of C. argenteus showed high activity on germination and growth, and these effects take place by means of mitochondrial metabolism alterations and increase the oxidative stress, leading the seedling death.

Assessment of Volatile Chemical Composition of the Essential Oil of Jatropha ribifolia (Pohl) Baill by HS-SPME-GC-MS Using Different Fibers.

The chemical composition of essential oil and volatile obtained from the roots of Jatropha ribifolia (Pohl) Baill was performed in this work. The Clevenger extractor was utilized in hydrodistillation of oil and chemical composition determined by gas chromatography coupled with mass spectrometry detector (GC-MS). The identification of compounds was confirmed by retention index (Kovats index) obtained from a series of straight chain alkanes (C7-C30) and by comparison with NIST and ADAMS library. A total of 61 compounds were identified in essential oil by GC-MS. The extraction of volatile was performed also by the use of the solid phase microextraction (SPME) with four different fibers. The essential oil extraction was extremely rapid (15 s) to avoid saturation of the fiber and the MS detector. The majority of the composition of essential oil is the terpenes: ß-pinene (major compound 9.16%), ß-vatirene (8.34%), α-gurjunene (6.98%), α-pinene (6.35%), camphene (4.34%), tricyclene (3.79%) and dehydro aromadendrene (3.52%) it and aldehydes and alcohols. Through the SPME it was possible to determine the nine volatile compounds not identified in oil 2,3,4-trimethyl-2-cyclopenten-1-one, α-phellandrene, 3-carene, trans-p-mentha-2,8-dienol, pinocamphone, D-verbenon, 1,3,3-trimethyl-2-(2-methyl-cyclopropyl)-cyclohexene, 2,4-diisocyanato-1-methylbenzene, and (6-hydroxymethyl-2,3-dimethylehenyl) methanol.

Atividade fitotóxica de Croton doctoris S. Moore / Phytotoxic activity of Croton doctoris S. Moore

O objetivo deste trabalho foi caracterizar o efeito fitotóxico do extrato etanólico bruto e frações semipurificadas do caule e folhas de C. doctoris na germinação, crescimento inicial e divisão celular das células radiculares de alface (Lactuca sativa) e cebola (Allium cepa), em laboratório. O experimento foi realizado em delineamento inteiramente casualizado, sendo utilizadas quatro concentrações dos tratamentos (0, 250, 500, 1.000mg L-1), com quatro repetições. O extrato bruto e as frações do caule e folhas inibiram a germinação e crescimento de alface e cebola, sendo o maior efeito fitotóxico promovido pela fração acetato de etila (FAE) do caule e folhas que afetou significativamente o crescimento e índice mitótico das células radiculares de alface e cebola. Com os resultados obtidos, verifica-se que a espécie C. doctoris apresenta substâncias capazes de influenciar no crescimento e divisão celular das espécies avaliadas.

The aim of this study was to verify the phytotoxic effect of the crude ethanol extract and semipurified fractions from the stem and leaves of Croton doctoris on the germination, growth and cell division of roots cells of lettuce (Lactuca sativa) and onion (Allium cepa), in the laboratory. The bioassays was conducted in using completely randomized design, with four concentrations (0, 250, 500, 1.000mg L-1) and four replications. The crude extract and the fractions from the stem and leaves inhibited germination and seedling growth of lettuce and onion, with greater phytotoxic effect observed in the ethyl acetate fraction (EAF) from the stems and leaves that significantly affected the growth and mitotic index of root cells of lettuce and onions. With these results it appears that C. doctoris contains substances capable of influencing the germination, growth and cell division of the species studied.

Volatile compounds of leaves and fruits of Mangifera indica var. coquinho (Anacardiaceae) obtained using solid phase microextraction and hydrodistillation.

Volatile compounds present on fruits and leaves of Mangifera indica var. coquinho were investigated by headspace solid phase microextraction (HS-SPME) and hydrodistillation (HD). Conventional techniques, such as hydrodistillation, may impart chemical changes to the original oil composition being also time-consuming. On the other hand, HS-SPME provides solvent-less extractions, shorter extraction times and may supply complementary information about the composition of the compounds. The HS-SPME technique was previously evaluated by the comparative study among the fibres: commercial PDMS, NiTi-ZrO(2) and NiTi-ZrO(2)-PDMS. The fibre NiTi-ZrO(2)-PDMS showed better sensitivity and precision and was used on the extraction of components. The influence of several parameters like the time and temperature of extraction and desorption time were examined to obtain better efficiency. Fruits and leaves were analysed in mature and immature stages. The profiles corresponding to the volatile compounds detected by both techniques are discussed.

Essential oil from Zanthoxylum hyemale.

The essential oils from aerial parts of young (sample A) and mature leaves (sample B), fruit (sample C), and flowers (sample D) of Zanthoxylum hyemale were obtained by hydrodistillation and analyzed by GC, GC/MS, and chiral phase gas chromatography (CPGC). Thirty-four compounds were identified from the essential oils, representing approximately 90.71, 91.19, 87.33, and 89.08 % of the oils, respectively. The major constituent of the young leaf essential oil was the sesquiterpene trans-nerolidol (51 %), while the main constituent of mature leaf (31 %) and flower oils (22 %) was an as yet unknown humulane-type sesquiterpenoid, which was characterized by spectral techniques (EI-MS and 1D-, 2D-NMR) as 3,7,10,10-tetramethylcycloundeca-3,7-dien-1-ol ( 1) and given the trivial name "hyemalol". In the fruit essential oil, the most abundant components were the monoterpenes beta-pinene (25 %) and alpha-pinene (10 %). The antimicrobial activity of the essential oils and some isolated compounds is also reported.

Composition and antibacterial activity of the essential oils from Zanthoxylum rhoifolium.

The essential oils from the aerial parts of leaves, fruits and flowers of Zanthoxylum rhoifolium of Southern Brazil (Rio Grande do Sul), were analysed by GC, GC/MS, and chiral phase gas chromatography (CPGC). Forty-eight compounds were identified from the essential oils. The major constituents of the essential oil of the leaves were germacrene D (34 %) and bicyclogermacrene (23 %) and of the fruits, menth-2-en-1-ol (46.2 %), beta-myrcene (30.2 %), (-)-linalool (15 %) and (-)-alpha-terpineol (8.45 %). beta-Myrcene (65 %) and menth-2-en-1-ol (5.4 %) dominate the essential oil of the flowers. The oils of the leaves and fruits were bioactive with antibacterial activity against Staphylococcus aureus (Gram positive), and Klebsiella pneumoniae and Salmonella setubal bacteria (Gram negative) microorganisms, while the essential oil of the flowers was inactive.

Chemical analysis and antifungal activity of the essential oil of Calea clematidea.

The chemical composition of the essential oils of Calea clematidea Baker obtained by hydrodistillation of the leaves and flowers was analysed by GC and GC/MS and the oils were assayed for their antifungal activities. The essential oil of the leaves showed a high content of a new natural epoxy terpenoid, named clemateol (ca. 70 %), with minor amounts of o-vanillin (6.5 %), spathulenol (4.2 %), alpha-terpinene (4.0 %), germacrene B (2.9 %), yomogi alcohol (1.8 %), ( E)-caryophylene (1.7 %), m-cymenene (1.6 %), and alpha-gurjunene (1.5 %), while the essential oil of the flowers was characterized by a higher content of thymol methyl ether (ca. 80 %), with minor amounts of clemateol (4.8 %) and o-cymene (4.7 %). The antimicrobial activity of the oils was also evaluated against dermatophytes for their possible use in pharmaceutical preparations for topical applications. The oil of the leaves (MIC > 3.57 mg/ml), clemateol (MIC > 1.52 mg/ml), and the alcohol 2 (MIC > 2.82 mg/ml) showed a moderate antifungal activity against Trichophyton tonsurans, Trichophyton rubrum, Trichophyton menthagrophytes var. i nterdigitale, Epidermophyton floccosum, Microsporum gypseum, Microsporum canis and Microsporum nanum.

Quinoline alkaloids, coumarins and volatile constituents of Helietta longifoliata.

A new quinoline alkaloid named helietidine ( 1), and seven known compounds ( 2 - 8) have been isolated from the stem barks of Helietta longifoliata. The structures of the new and the known compounds were established on the basis of spectral evidence, especially by 2D NMR ( 1H- 1H COSY, NOESY, HMQC, HMBC). In addition, the volatile constituents of H. longifoliata leaves were analysed by gas chromatography-mass spectrometry (GC-MS). Twenty-five constituents were identified representing ca. of 96 % of the oil, and limonene (17.50 %), germacrene D (16.60 %), elemol (11.81 %), bicyclogermacrene (11.67 %), guaiol (11.53), and epi-alpha-bisabolol (7.24 %) were the most abundant components. The oil was bioactive against some Gram-positive and Gram-negative bacteria, as revealed by bioautography.