Repellency effect of Pilocarpus spicatus A. St.-Hil essential oil and nanoemulsion against Rhipicephalus microplus larvae.

Rhipicephalus microplus, the cattle tick, is a major cause of economic losses in bovine production. Due to the widespread acaricidal resistance to commercially available products, as well as their toxicity and environmental impact, alternative control methods are required. Nanoformulations produced from plant extracts as bioactive substances are very promising as innovative acaricidal agents. Thus, the aim of this study was to evaluate the in vitro repellent activity of Pilocarpus spicatus essential oil and its nanoemulsion against R. microplus, using larval repellent test (RT). The essential oil was extracted by hydrodistillation, using a Clevenger-type apparatus. The nanoemulsion was prepared with 5% essential oil, 5% tween 80, and 90% water, using the phase inversion method (50 mg/mL). Limonene was the major component (46.8%) of the essential oil, as determined by gas chromatography-mass spectrometry (GC/MS) and confirmed by flame ionization detection (GC/FID). According to the RT results, the essential oil had a repellent activity greater than 69%, from concentrations of 3.12 mg/mL (69.81 ± 10%) to 50 mg/mL (98.10 ± 0.6%), whereas the nanoemulsion at 50 mg/mL presented repellent activities of 97.14 ± 1.37% and 97.89 ± 0.52% 6 and 10 h after treatment, respectively. These values regarding to total repellency were very close to those calculated for mortality corrected by Abbott's formula. The phase inversion method preserved the chemical and physical characteristics of the essential oil since both reached an equal repellent effect at the same concentration. Therefore, P. spicatus essential oil and nanoemulsion had excellent repellent activities against R. microplus larvae, demonstrating its potential for future use as an alternative for tick control.

Preparation of a Nanoemulsion with Carapa guianensis Aublet (Meliaceae) Oil by a Low-Energy/Solvent-Free Method and Evaluation of Its Preliminary Residual Larvicidal Activity.

Andiroba (Carapa guianensis) seeds are the source of an oil with a wide range of biological activities and ethnopharmacological uses. However, few studies have devoted attention to innovative formulations, including nanoemulsions. The present study aimed to obtain a colloidal system with the andiroba oil using a low-energy and organic-solvent-free method. Moreover, the preliminary residual larvicidal activity of the nanoemulsion against Aedes aegypti was evaluated. Oleic and palmitic acids were the major fatty acids, in addition to the phytosterol ß-sitosterol and limonoids (tetranortriterpenoids). The required hydrophile-lipophile was around 11.0 and the optimal nanoemulsion was obtained using polysorbate 85. The particle size distribution suggested the presence of small droplets (mean diameter around 150 nm) and low polydispersity index (around 0.150). The effect of temperature on particle size distribution revealed that no major droplet size increase occurred. The preliminary residual larvicidal assay suggested that the mortality increased as a function of time. The present study allowed achievement of a potential bioactive oil in water nanoemulsion that may be a promising controlled release system. Moreover, the ecofriendly approach involved in the preparation associated with the great bioactive potential of C. guianensis makes this nanoemulsion very promising for valorization of this Amazon raw material.

Development of a larvicidal nanoemulsion with Copaiba (Copaifera duckei) oleoresin

Copaiba (Copaifera duckei Dwyer, Fabaceae) oleoresin is an important Amazonian raw material. Despite its insecticidal potential, poor water solubility remains a challenge for the development of effective and viable products. Nanotechnology has emerged as a promising area to solve this problem, especially oil-in-water nanoemulsions. On this context, the aim of the present study was to develop oil-in-water nanoemulsions using copaiba oleoresin dispersed through a high internal phase; and evaluate its potential insecticidal action against Aedes aegypti larvae. Overall, 31 formulations were prepared, ranging from 11.5 ± 0.2 to 257.3 ± 4.1 nm after one day of manipulation. Some of them reached small mean droplet sizes (< 200 nm) and allowed achievement of a nanoemulsion region. The formulation consisted of 5% (w/w) of copaiba oil, 5% (w/w) of surfactant and 90% (w/w) of water, which presented mean droplet size of 145.2 ±0.9 nm and polidispersity of 0.378 ± 0.009 after one day of manipulation, and these were evaluated for larvicidal potential. According to mortality level (250 ppm - 93.3 after 48 h), this nanoemulsion was classified as a promising insecticidal agent against Aedes aegypti larvae. The present study allowed the development of low-cost ecofriendly green natural-based nanoformulations with potential larvicidal activity, using a nanobiotechnology approach.

Triterpene esters and biological activities from edible fruits of Manilkara subsericea (Mart.) Dubard, Sapotaceae.

Manilkara subsericea (Mart.) Dubard (Sapotaceae) is popularly known in Brazil as "guracica." Studies with Manilkara spp indicated the presence of triterpenes, saponins, and flavonoids. Several activities have been attributed to Manilkara spp such as antimicrobial, antiparasitic and antitumoral, which indicates the great biological potential of this genus. In all, 87.19% of the hexanic extract from fruits relative composition were evaluated, in which 72.81% were beta- and alpha-amyrin esters, suggesting that they may be chemical markers for M. subsericea. Hexadecanoic acid, hexadecanoic acid ethyl ester, (E)-9-octadecenoic acid ethyl ester, and octadecanoic acid ethyl ester were also identified. Ethanolic crude extracts from leaves, stems, and hexanic extract from fruits exhibited antimicrobial activity against Staphylococcus aureus ATCC25923. These extracts had high IC50 values against Vero cells, demonstrating weak cytotoxicity. This is the first time, to our knowledge, that beta- and alpha-amyrin caproates and caprylates are described for Manilkara subsericea.

Antiproliferative activity of Eremanthus crotonoides extracts and centratherin demonstrated in brain tumor cell lines

The genus Eremanthus is recognized by the predominance of sesquiterpene lactones from the furanoheliangolide type, a class of substances extensively tested against cancer cell lines. Thus, the species E. crotonoides (DC.) Sch. Bip., Asteraceae, obtained on "restinga" vegetation was evaluated against U251 and U87-MG glioma cell lines using the MTT colorimetric assay. Dichloromethane fraction was cytotoxic to both glioblastoma multiforme cell lines. We then conducted UPLC-PDA-ESI-MS/MS analysis of the dichloromethane fraction, which allowed the identification of the sesquiterpene lactones centratherin and goyazensolide. The isolation of centratherin was performed using chromatographic techniques and the identification of this substance was confirmed according to NMR data. Cytotoxic activity of centratherin alone was also evaluated against both U251 and U87-MG cells, which showed IC50 values comparable with those obtained for the commercial anticancer drug doxorubicin. All the tested samples showed cytotoxic activity against glioblastoma multiforme cells which suggests that E. crotonoides extracts may be important sources of antiproliferative substances and that the centratherin may serve as prototype for developing new antiglioblastoma drugs.

Atypical polycystic echinococcosis without liver involvement in Brazilian patients.

Echinococcus vogeli is known to occur in the neotropics, causing polycystic echinococcosis (PE) in humans. The liver and lungs are the most commonly affected organs. In this paper, we report six cases of human PE in the mesenteries without primary liver involvement. All patients, four males and two females (median age 34 years; range 20-48 years), lived in a forested area (Acre, Brazil). Physical examination revealed abdominal masses. The patients were evaluated by abdominal CT scan, exploratory laparotomy and serology by immunoblot assay. CT scans revealed multiple and cystic lesions with calcified edges in the mesenteric segments. During exploratory laparotomy, polycystic lesions were excised from omentum, mesentery, pancreas and inferior vena cava. Positive serology identified 21 and 31 kDa subunits. We recommend that PE should be investigated in patients from tropical forests in northern Brazil exhibiting abdominal masses with no clearly defined diagnosis.

Intraspecific variation of Echinococcus granulosus in livestock from Peru.

Echinococcus granulosus infection is a problem of public health in livestock-rearing regions of Latin America, like in Peru. This species is characterized by the morphometry of the large and small rostellar hooks in both metacestodes and adult worms. The aim of this work was to study biometrical characteristics of rostellar hooks in hydatid cysts (lung and liver) from some Peruvian provinces (Arequipa, Cuzco, Huancavelica, Junin and Puno). The biometrical analysis was obtained in a digital system for image analysis. The following parameters were studied: area, perimeter, length and width total of the handle, the blade and the distance between the blade and the guard of large and small hooks. For statistical analysis Student's t-test and Analysis of Variance (ANOVA) with Pos-test of Tukey were used, considering the values of p< or =0.05 as of statistic significance. All morphometric characters of large and small hooks were highly significant (p<0.000), when data were compared among the samples from the different provinces. Rostellar hooks in specimens from Huancavelica presented smallest measurements, whereas hooks in specimens from Junin were bigger in relation to the total and blade measurements. Generally, samples from Arequipa presented the biggest large hooks in relation to the handle parameters. The rostellar hooks of cattle and sheep hosts varied depending on the geographic localization, host and organ. Hooks recovered from the lungs were larger than those collected in the liver. It was concluded that morphometry using computer image analysis allows the characterization of E. granulosus. The rostellar hooks presented morphometric polymorphism.