a b s t r a c tNanotechnology has emerged as a promising area for innovative products, including insecticides. Dengue is a tropical disease which is considered a critical health problem in developing countries, due to negative impacts to the environment caused by synthetic chemicals used for vector control (Aedes aegypti). Thus, developing of natural products based insecticidal are considered very promising. On this context, the aim of the present study was to obtain an O/W nanoemulsion containing Rosmarinus officinalis L., Lamiaceae, essential oil and evaluate its larvicidal activity against A. aegypti. Low energy method was employed, allowing achievement of small droplets. The nanoemulsion also presented low polydispersity and mean droplet below 200 nm, even after 30 days of storage. Potential mortality levels were observed after 24 h (80 ± 10%) and 48 h (90 ± 10%) in A. aegypti larvae at final concentration of 250 ppm, related to R. officinalis essential oil. This study contributes to nanobiotechnology of natural products, presenting a potential larvicidal nanoemulsion prepared with R. officinalis essential oil. Moreover, nanoemulsion production involved a non-heating procedure, describing easy technique which may be useful for integrative control programs.

CopaibaLarvicidal Nanoemulsion Oleoresin A B S T R A C T 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.

Pterodon emarginatus Vogel is a Brazilian species that belongs to the family Fabaceae, popularly known as sucupira. Its oil has several biological activities, including potent larvicidal property against Aedes aegypti. This insect is the vector of dengue, a tropical disease that has been considered a critical health problem in developing countries, such as Brazil. Most of dengue control methods involve larvicidal agents suspended or diluted in water and making active lipophilic natural products available is therefore considered a technological challenge. In this context, nanoemulsions appear as viable alternatives to solve this major problem. The present study describes the development of a novel nanoemulsion with larvicidal activity against A. aegypti along with the required Hydrophile Lipophile Balance determination of this oil. It was suggested that the mechanism of action might involve reversible inhibition of acetylcholinesterase and our results also suggest that the P. emarginatus nanoemulsion is not toxic for mammals. Thus, it contributes significantly to alternative integrative practices of dengue control, as well as to develop sucupira based nanoproducts for application in aqueous media.

Baccharis reticularia DC. is a plant species from the Asteraceae family that is endemic to Brazil. Despite the great importance of Baccharis genus, no study has been carried out regarding either the phytochemical composition of B. reticularia or the evaluation of its larvicidal potential. Considering the intrinsic immiscibility of essential oils, this study shows larvicidal nanoemulsions containing the B. reticularia phytochemically characterized essential oil and its main constituent against Aedes aegypti. The major compound found was d-limonene (25.7%). The essential oil inhibited the acetylcholinesterase, one of the main targets of insecticides. The required hydrophile-lipophile balance of both nanoemulsions was 15.0. The mean droplet sizes were around 90.0 nm, and no major alterations were observed after 24 h of preparation for both formulations. After 48 h of treatment, the estimated LC50 values were 118.94 μg mL−1 and 81.19 μg mL−1 for B. reticularia essential oil and d-limonene nanoemulsions, respectively. Morphological alterations evidenced by scanning electron micrography were observed on the larvae treated with the d-limonene nanoemulsion. This paper demonstrated a simple and ecofriendly method for obtaining B. reticularia essential oil and d-limonene aqueous nanoemulsions by a non-heating and solvent-free method, as promising alternatives for Aedes aegypti control.

BackgroundPreparation of nanoformulations using natural products as bioactive substances is considered very promising for innovative larvicidal agents. On this context, oil in water nanoemulsions develop a main role, since they satisfactorily disperse poor-water soluble substances, such as herbal oils, in aqueous media. Pterodon emarginatus, popularly known as sucupira, has a promising bioactive oleoresin. However, to our knowledge, no previous studies were carried out to evaluate its potential against Culex quinquefasciatus, the main vector of the tropical neglected disease called lymphatic filariasis or elephantiasis. Thus, we aimed to investigate influence of different pairs of surfactants in nanoemulsion formation and investigate if a sucupira oleoresin-based nanoemulsion has promising larvicidal activity against this C. quinquefasciatus. We also evaluated morphological alteration, possible mechanism of insecticidal action and ecotoxicity of the nanoemulsion against a non-target organism.ResultsAmong the different pairs of surfactants that were tested, nanoemulsions obtained with polysorbate 80/sorbitan monooleate and polysorbate 80/sorbitan trioleate presented smallest mean droplet size just afterwards preparation, respectively 151.0 ± 2.252 and 160.7 ± 1.493 nm. They presented high negative zeta potential values, low polydispersity index (<0.300) and did not present great alteration in mean droplet size and polydispersity index after 1 day of preparation. Overall, nanoemulsion prepared with polysorbate 80/sorbitan monooleate was considered more stable and was chosen for biological assays. It presented low LC50 value against larvae (34.75; 7.31–51.86 mg/L) after 48 h of treatment and some morphological alteration was observed. The nanoemulsion did not inhibit acetylcholinesterase of C. quinquefasciatus larvae. It was not toxic to green algae Chlorella vulgaris at low concentration (25 mg/L).ConclusionsOur results suggest that optimal nanoemulsions may be prepared with different surfactants using a low cost and low energy simple method. Moreover, this prototype proved to be effective against C. quinquefasciatus, being considered an ecofriendly novel nanoproduct that can be useful in integrated control programs of vector control.

Outbreaks of diseases pose a major threat to sustainable aquaculture development worldwide. Application of herbal products to combat parasitic diseases provides an alternative approach for sustainable aquaculture. This study investigated the in vitro antiparasitic effects of an oil-in-water nanoemulsion prepared using the essential oil from Pterodon emarginatus, against monogeneans infesting Colossoma macropomum. Gill arches from C. macropomum (47.6 ± 14.5 g and 13.5 ± 1.4 cm) that were naturally parasitized by Anacanthorus spathulatus, Notozothecium janauachensis and Mymarothecium boegeri were immersed in different dispersions of the P. emarginatus nanoemulsions (0, 50, 100, 200, 400 and 600 mg/L). The major compounds presented in the essential oil of P. emarginatus were β-elemene, β-caryophyllene and α-humulene. Characterization of these nanoemulsions showed that they have a small mean droplet size and low polydispersity index, which is concordant with stable systems. In this in vitro trial, the P. emarginatus nanoemulsion concentrations of 100, 200, 400 and 600 mg/L presented 100% helminthic efficacy against monogeneans of the gills of C. macropomum. The highest two concentrations used (400 and 600 mg/L) were seen to immobilize the parasites after only 15 min. Therefore, it would be worthwhile testing these concentrations in therapeutic baths against monogeneans of C. macropomum.

Our results suggest that the acute administration of the HELSm in the scototaxis and NTDT tests in a zebrafish model (Danio rerio) produced anxiolytic and antidepressant effects, devoid of hypnotic and sedative actions by immersion, and this action was improved when administered by oral via. Possibly, the presence of isoquercitrin in the leaves of Spondias mombin participates in the anxiolytic and antidepressant effects.

Monogeneans are ectoparasites that may cause losses in production and productivity in the aquaculture of Colossoma macropomum. Chemotherapeutics used in aquaculture usually have major adverse effects on fish; hence, the use of essential oils has been considered advantageous, but these are not soluble in water. Thus, the use of nanostructures to enhance water solubility of compounds and improve bioactivity may be very promising. This study investigated the antiparasitic activity of nanoemulsion prepared with Copaifera officinalis oleoresin (50, 100, 150, 200 and 300 mg/L), against monogenean parasites from the gills of C. macropomum. The particle size distribution and zeta potential suggested that a potentially kinetic stable system was generated. The nanoemulsion from C. officinalis oleoresin achieved high efficacy (100%) at low concentrations (200 and 300 mg/L) after 15 min of exposure. This was the first time that a nanoemulsion was generated from C. officinalis oleoresin using a solvent-free, non-heating and low-energy method. Moreover, this was the first time that an antiparasitic against monogeneans on fish gills, based on nanoemulsion of C. officinalis oleoresin, was tested.