Antileishmanial activity of diterpene acids in copaiba oil

Leishmaniasis is a neglected tropical disease. According to the World Health Organization, there are approximately 1.5-two million new cases of cutaneous leishmaniasis each year worldwide. Chemotherapy against leishmaniasis is based on pentavalent antimonials, which were developed more than a century ago. The goals of this study were to investigate the antileishmanial activity of diterpene acids in copaiba oil, as well as some possible targets of their action against Leishmania amazonensis. Methyl copalate and agathic, hydroxycopalic, kaurenoic, pinifolic and polyaltic acids isolated from Copaifera officinales oleoresins were utilised. Ultrastructural changes and the specific organelle targets of diterpenes were investigated with electron microscopy and flow cytometry, respectively. All compounds had some level of activity against L. amazonensis. Hydroxycopalic acid and methyl copalate demonstrated the most activity against promastigotes and had 50% inhibitory concentration (IC50) values of 2.5 and 6.0 µg/mL, respectively. However, pinifolic and kaurenoic acid demonstrated the most activity against axenic amastigote and had IC50 values of 3.5 and 4.0 µg/mL, respectively. Agathic, kaurenoic and pinifolic acid caused significant increases in plasma membrane permeability and mitochondrial membrane depolarisation of the protozoan. In conclusion, copaiba oil and its diterpene acids should be explored for the development of new antileishmanial drugs.

The use of halloysite clay and carboxyl-functionalised multi-walled carbon nanotubes for recombinant LipL32 antigen delivery enhanced the IgG response

We studied the feasibility of using halloysite clay nanotubes (HNTs) and carboxyl-functionalised multi-walled carbon nanotubes (COOH-MWCNTs) as antigen carriers to improve immune responses against a recombinant LipL32 protein (rLipL32). Immunisation using the HNTs or COOH-MWCNTs significantly increased the rLipL32-specific IgG antibody titres (p < 0.05) of Golden Syrian hamsters. None of the vaccines tested conferred protection against a challenge using a virulent Leptospira interrogans strain. These results demonstrated that nanotubes can be used as antigen carriers for delivery in hosts and the induction of a humoral immune response against purified leptospiral antigens used in subunit vaccine preparations.

Antileishmanial activity of the essential oil from Tetradenia riparia obtained in different seasons

The herbaceous shrub Tetradenia riparia has been traditionally used to treat inflammatory and infectious diseases. Recently, a study showed that T. riparia essential oil (TrEO) obtained in summer has antileishmanial effects, although these results could be influenced by seasonal variation. This study evaluated the activity of the TrEO obtained in different seasons against Leishmania (Leishmania) amazonensis, in vitro and in vivo. The compounds in the TrEO were analysed by gas chromatography-mass spectrometry; terpenoids were present and oxygenated sesquiterpenes were the majority compounds (55.28%). The cytotoxicity and nitric oxide (NO) production were also tested after TrEO treatment. The TrEO from all seasons showed a 50% growth inhibitory concentration for promastigotes of about 15 ng/mL; at 30 ng/mL and 3 ng/mL, the TrEO reduced intracellular amastigote infection, independently of season. The TrEO from plants harvested in summer had the highest 50% cytotoxic concentration, 1,476 ng/mL for J774.A1 macrophages, and in spring (90.94 ng/mL) for murine macrophages. NO production did not change in samples of the TrEO from different seasons. The antileishmanial effect in vivo consisted of a reduction of the parasite load in the spleen. These results suggest that the TrEO has potential effects on L. (L.) amazonensis, consonant with its traditional use to treat parasitic diseases.

The mc2-CMX vaccine induces an enhanced immune response against Mycobacterium tuberculosis compared to Bacillus Calmette-Guérin but with similar lung inflammatory effects

Although the attenuated Mycobacterium bovis Bacillus Calmette-Guérin (BCG) vaccine has been used since 1921, tuberculosis (TB) control still proceeds at a slow pace. The main reason is the variable efficacy of BCG protection against TB among adults, which ranges from 0-80%. Subsequently, the mc2-CMX vaccine was developed with promising results. Nonetheless, this recombinant vaccine needs to be compared to the standard BCG vaccine. The objective of this study was to evaluate the immune response induced by mc2-CMX and compare it to the response generated by BCG. BALB/c mice were immunised with both vaccines and challenged withMycobacterium tuberculosis (Mtb). The immune and inflammatory responses were evaluated by ELISA, flow cytometry, and histopathology. Mice vaccinated with mc2-CMX and challenged with Mtb induced an increase in the IgG1 and IgG2 levels against CMX as well as recalled specific CD4+ T-cells that produced T-helper 1 cytokines in the lungs and spleen compared with BCG vaccinated and challenged mice. Both vaccines reduced the lung inflammatory pathology induced by the Mtb infection. The mc2-CMX vaccine induces a humoral and cellular response that is superior to BCG and is efficiently recalled after challenge with Mtb, although both vaccines induced similar inflammatory reductions.

Antimicrobial activity of Eucalyptus globulus oil, xylitol and papain: a pilot study

OBJECTIVE To evaluate the in vitro antimicrobial activity of the Eucalyptus globulus essential oil, and of the xylitol and papain substances against the following microorganisms: Pseudomonas aeruginosa; Samonella sp.; Staphylococus aureus; Proteus vulgaris; Escherichia coli and Candida albicans. METHOD The in vitro antimicrobial evaluation was used by means of the agar diffusion test and evaluation of the inhibition zone diameter of the tested substances. Chlorhexidine 0.5% was used as control. RESULTS The Eucalyptus globulus oil showed higher inhibition than chlorhexidine when applied to Staphylococcus aureus, and equal inhibition when applied to the following microorganisms: Escherichia coli, Proteus vulgaris and Candida albicans. Papain 10% showed lower antimicrobial effect than chlorhexidine in relation to Candida albicans. Xylitol showed no inhibition of the tested microorganisms. CONCLUSION The Eucalyptus globulus oil has antimicrobial activity against different microorganisms and appears to be a viable alternative as germicidal agent hence, further investigation is recommended.

Females of Tapinotaspoides, a genus in the oil-collecting bee tribe Tapinotaspidini, collect secretions from non-floral trichomes (Hymenoptera, Apidae)

Harvesting of secretions from non-floral trichomes by females of Tapinotaspoides serraticornis is reported for the first time. The females exhibit a type of mopping behavior using the fringes of long, wavy setae along the posterior margins of their metasomal sterna. Our observations indicated a wide range of host plants used as sources for these secretions, including Waltheria (Sterculiaceae), Tibouchina (Melastomataceae), Sida (Malvaceae), Jacquemontia (Convolvulaceae), and unidentified species of Commelinaceae and Cyperaceae.

Notes on oil sources for the bee genus Caenonomada(Hymenoptera, Apidae, Tapinotaspidini)

It is reported for the first time oil collecting by bees of the genus Caenonomada on flowers of Plantaginaceae. Females of Caenonomada unicalcarata were observed collecting oil on flowers of Angelonia cornigera, and males and females of Caenonomada bruneri and C. aff. unicalcarata were observed on flowers of Angelonia and Monopera (Plantaginaceae). The record of Caenonomada on Plantaginaceae suggests the use of trichomatic oil glands as a primitive condition in the tribe Tapinotaspidini.

The effect of sub-lethal doses of Azadirachta indica (Meliaceae) oil on the midgut of Spodoptera frugiperda (Lepidoptera, Noctuidae)

The fall armyworm, Spodoptera frugiperda, is one of the major field pests for maize production. It is mainly controlled by means of synthetic, and more recently by resistant cultivar of maize expressing Bt toxins. The neem tree, Azadirachta indica, is a plant that can potentially control insects with the advantage of being food and environmental safe. The aim of this study was to assess the effect of neem oil on the development and survival of S. frugiperda caterpillars by assessing histological alterations caused on their midgut. Newly hatched caterpillars were submitted to three neem oil concentrations: 0.006; 0.05; 0.4%, which were added to their artificial diet. Ten 3rd instar caterpillars, taken from each treatment, were submitted to histological analysis. The alimentary canals from the specimens were fixed in Baker for 12 hours, desiccated and diaphanized in alcohol/xylol (1:1) and xylol. After placing the samples in paraffin, they were sliced in 8 µm sections and stained with hematoxylin-eosin stain. The neem oil added to the diet of S. frugiperda caused total mortality at dose of 0.4% whilst still in the first instars, prolonged the larval and pupal stages, and reduced the pupal weight. Histo-physiological alterations such as degeneration of the epithelial lining of the midgut and in the peritrophic matrix were found at all concentrations of neem oil.

Enhanced degradation of metalaxyl in Gley Humic and Dark Red Latosol

Enhanced degradation of the fungicide metalaxyl was investigated in two soils: a gley humic (GH) and a Dark Red Latosol (LE), collected at sites never exposed to the fungicide. The soil samples were treated with successive applications of metalaxyl as a commercial formulation and 14C-metalaxyl in laboratory. Metalaxyl biodegradation was analyzed during 63 days by means of radiometric techniques to verify biomineralization and degradation product formation from the applied 14C-metalaxyl. Although biomineralization (maximum of 14 and 8% in the GH and LE soils, respectively), and partial degradation (about 32 and 48%, respectively) were detected in both soils, enhanced degradation was verified only in the GH soil. Results proved that metalaxyl behaves differently in soils.

Molecular analysis of the bacterial diversity in a specialized consortium for diesel oil degradation

Diesel oil is a compound derived from petroleum, consisting primarily of hydrocarbons. Poor conditions in transportation and storage of this product can contribute significantly to accidental spills causing serious ecological problems in soil and water and affecting the diversity of the microbial environment. The cloning and sequencing of the 16S rRNA gene is one of the molecular techniques that allows estimation and comparison of the microbial diversity in different environmental samples. The aim of this work was to estimate the diversity of microorganisms from the Bacteria domain in a consortium specialized in diesel oil degradation through partial sequencing of the 16S rRNA gene. After the extraction of DNA metagenomics, the material was amplified by PCR reaction using specific oligonucleotide primers for the 16S rRNA gene. The PCR products were cloned into a pGEM-T-Easy vector (Promega), and Escherichia coli was used as the host cell for recombinant DNAs. The partial clone sequencing was obtained using universal oligonucleotide primers from the vector. The genetic library obtained generated 431 clones. All the sequenced clones presented similarity to phylum Proteobacteria, with Gammaproteobacteria the most present group (49.8 % of the clones), followed by Alphaproteobacteira (44.8 %) and Betaproteobacteria (5.4 %). The Pseudomonas genus was the most abundant in the metagenomic library, followed by the Parvibaculum and the Sphingobium genus, respectively. After partial sequencing of the 16S rRNA, the diversity of the bacterial consortium was estimated using DOTUR software. When comparing these sequences to the database from the National Center for Biotechnology Information (NCBI), a strong correlation was found between the data generated by the software used and the data deposited in NCBI.

Recovery of degraded pasture in Rondônia: macronutrients and productivity of brachiaria brizantha

Pasture is the main form of land use in Amazonia. Over time the pasture grass loses vigor and yields decrease, indicating a certain degree of degeneration. The main causes of degradation are lack of pasture maintenance and subsequent weed infestation, the choice of regionally unsuitable forage species and excessive grazing. The main purpose of this study was to evaluate the impact of different recovery managements on soil chemical properties and grass yield of a degraded pasture in Rondônia. For this purpose, an experiment was installed in October 2001, consisting of five treatments: C = control; HA = harrowing + NPK + micronutrients; HE = Herbicide + NK + micronutrients; R = No-tillage rice + NPK + micronutrients; and S = No-tillage soybean + PK + micronutrients. The following N, P and K sources were used: ammonium sulfate for N, calcined phosphate for P and potassium chloride for K. The experiment was arranged in a randomized block design with four replications. The shoot dry matter yield of the grass was analyzed as of the 35th month of experimentation, in a dry and a rainy period. Phosphorus fertilization resulted in significant increases in Ca2+ and Mg2+ and increasing trend of P in the topsoil in the initial months of the experiment in treatments HA and S and increases in Ca2+ and P (trend) in the treatment R. The cumulative production of Brachiaria brizantha, from Sep/2004 to Mar/2005, was 30,025, 28,267 and 27,735 kg ha-1 shoot dry matter in the treatments HA, R and S, respectively. These values differed significantly from treatments C and HE, with 17,040 and 17,057 kg ha-1, respectively. It was concluded that phosphorus fertilization associated to pasture reform was effective to raise the dry matter yield of Brachiaria brizantha. Rice or soybean under no-tillage is recommended as a practice of pasture recovery, due to the residual effect of fertilization.

Nitrogen fertilizer use efficiency, recovery and leaching of an alexandergrass pasture

Nitrogen usually determines the productive potential of forage crops, although it is highly unstable in the environment. Studies on recovery rates and use efficiency are important for more reliable fertilizer recommendations to reduce costs and avoid environmental pollution. The purpose of this study was to evaluate N use efficiency and recovery rate of Alexandergrass pasture (Brachiaria - Syn. Urochloa plantaginea) as well as N-NO3- and N-NH4+ soil concentrations using different levels of N fertilization under two grazing intensities. The experiment was arranged in a randomized block design in a factorial scheme with three replications. Treatments consisted of three N rates (0, 200 and 400 kg ha-1 N) and two grazing intensities termed low mass (LM; forage mass of 2,000 kg ha-1 of DM) and high mass (HM; forage mass of 3,600 kg ha-1 of DM) under continuous stocking and variable stocking rates. Results of N fertilization with 200 kg ha-1 were better than with 400 kg ha-1 N. There was a significant effect of N rates on soil N-NO3-concentration with higher levels in the first layer of the soil profile in the treatment with 400 kg ha-1 N. Grazing intensity also affected soil N-NO3- concentration, by increasing the levels under the higher stocking rate (lower forage mass).

Recovery of soil physical properties by green manure, liming, gypsum and pasture and spontaneous native species¹

Inadequate usage can degrade natural resources, particularly soils. More attention has been paid to practices aiming at the recovery of degraded soils in the last years, e.g, the use of organic fertilizers, liming and introduction of species adapted to adverse conditions. The purpose of this study was therefore to investigate the recovery of physical properties of a Red Latosol (Oxisol) degraded by the construction of a hydroelectric power station. In the study area, a soil layer about 8m thick had been withdrawn by heavy machines leading not only to soil compaction, but resulting in high-degree degradation. The experiment was arranged in a completely randomized design with nine treatments and four replications. The treatments consisted of: 1- soil mobilization by tilling (to ensure the effect of mechanical mobilization in all treatments) without planting, but growth of spontaneous vegetation; 2- Black velvet bean (Stizolobium aterrimum Piper & Tracy); 3- Pigeonpea (Cajanus cajan (L.) DC); 4- Liming + black velvet bean; 5-Liming + pigeonpea until 1994, when replaced by jack bean (Canavalia ensiformis); 6- Liming + gypsum + black velvet bean; 7- Liming + gypsum + pigeonpea until 1994, when replaced by jack bean; and two controls as reference: 8- Native Cerrado vegetation and 9- bare soil (no tilling and no planting), left under natural conditions and in this situation, without spontaneous vegetation. In treatments 1 through 7, the soil was tilled. Treatments were installed in 1992 and left unmanaged for seven years, until brachiaria (Brachiaria decumbens) was planted in all plots in 1999. Seventeen years after implantation, the properties soil macroporosity, microporosity, total porosity, bulk density and aggregate stability were assessed in the previously described treatments in the soil layers 0.00-0.10; 0.10-0.20 and 0.20-0.40 m, and soil Penetration Resistance and soil moisture in 0.00-0.15 and 0.15-0.30 m. The plants were evaluated for: brachiaria dry matter and spontaneous growth of native tree species in the plots as of 2006. Results were analyzed by variance analysis and Tukey´s test at 5 % for mean comparison. In all treatments, except for the bare soil (no recovery measures), ongoing recovery of the degraded soil physical properties was observed. Macroporosity, soil bulk density and total porosity were good soil quality indicators. The occurrence of spontaneous native species indicated the soil recovery process. The best adapted species was Machaerium acutifolium Vogel, with the largest number of plants and most advanced development; the dry matter production of B. decumbens in recovering soil was similar to normal conditions, evidencing soil recovery.

Diagnosis and recommendation integrated system (DRIS) of soybean seed oil content

The Diagnosis and Recommendation Integrated System (DRIS) can improve interpretations of leaf analysis to determine the nutrient status. Diagnoses by this method require DRIS norms, which are however not known for oil content of soybean seeds. The aims of this study were to establish and test the DRIS method for oil content of soybean seed (maturity group II cultivars). Soybean leaves (207 samples) in the full flowering stage were analyzed for macro and micro-nutrients, and the DRIS was applied to assess the relationship between nutrient ratios and the seed oil content. Samples from experimental and farm field sites of the southernmost Brazilian state Rio Grande do Sul (28° - 29° southern latitude; 52° -53° western longitude) were assessed in two growing seasons (2007/2008 and 2008/2009). The DRIS norms related to seed oil content differed between the studied years. A unique DRIS norm was established for seed oil content higher than 18.68 % based on data of the 2007/2008 growing season. Higher DRIS indices of B, Ca, Mg and S were associated with a higher oil content, while the opposite was found for K, N and P. The DRIS can be used to evaluate the leaf nutrient status of soybean to improve the seed oil content of the crop.