Nanoencapsulated Melaleuca alternifolia essential oil exerts anesthetic effects in the brachyuran crab using Neohelice granulate.

The aim of this study was to evaluate the efficacy and safety of several anesthetics in the brachyuran crab Neohelice granulata, an emergent experimental model. The essential oils (EOs) of Lippia alba, Aloysia tryphilla, and Melaleuca alternifolia (tea tree oil; TTO), the isolated compounds eugenol, menthol, terpinen-4-ol, and the nanoencapsulated form of TTO, were administered in one or more of the following ways: added to the water (immersion), through an arthrodial membrane (injected), or by oral gavage. Unexpectedly, most EOs did not produce an anesthetic effect after immersion. Only TTO and eugenol induced anesthesia by immersion, with very long induction and recovery times compared to anesthesia of other crustaceans. However, a good anesthetic effect was observed with the injection of terpinen-4-ol and nanoencapsulated TTO in N. granulata; both demonstrated ideal induction and recovery times. These substances appear to be promising anesthetic alternatives for crustaceans.

Melaleuca alternifolia anthelmintic activity in gerbils experimentally infected by Haemonchus contortus.

Gastrointestinal parasites are one of the biggest health problems faced in sheep, mainly due to their pathogenicity and resistance to drugs used to control these parasites. Thus, the following study aimed to assess the anthelmintic efficacy of Melaleuca alternifolia against Haemonchus contortus in gerbils (Meriones unguiculatus) experimentally infected. Three treatments were tested: M. alternifolia essential oil, popularly known as tea tree oil (TTO), a solid lipid nanocarrier made with essential oil of Melaleuca (nanoTTO), and terpinen-4-ol (terp-4-ol). In vivo studies were performed by determining the mean worm burden of H. contortus in gerbils TTO (0.75 mL/kg); nanoTTO (0.5 mL/kg) and terp-4-ol (0.5 mL l/kg) were able to reduce 46.36%; 48.64%, and 43.18% worm burden, respectively. H. contortus increased alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, as demonstrated by liver injury. It was found that the TTO, nanoTTO, and terp-4-ol were not toxic to liver and kidneys since hepatic and renal functions were not affected. Moreover, terp-4-ol was able to prevent increased levels of seric AST and ALT in infected animals, indicating a hepatoprotective effect. Thus, our results indicate that TTO, nanoTTO, and terp-4-ol are safe and efficient against H. contortus infection in gerbils, and possibly the terp-4-ol may be considered the compound present in the Melaleuca alternifolia responsible for parasitic action against H. contortus.

In vivo bactericidal effect of Melaleuca alternifolia essential oil against Aeromonas hydrophila: Silver catfish (Rhamdia quelen) as an experimental model.

Aeromonas hydrophila is one of the main causative agent of high mortality and significative economic losses in aquaculture and has become increasingly resistant to conventional antibiotics. One feasible alternative to control and treat it is the use of essential oils. This study aimed to evaluate A. hydrophila susceptibility to tea tree oil (TTO-Melaleuca alternifolia) in vivo, and the effect of this treatment. In vivo tests were performed using silver catfish (Rhamdia quelen) as the experimental model. Silver catfish were treated with TTO at 25 and 50 µL/L for seven days before infection. After seven days, the fish were inoculated with A. hydrophila via intramuscularly. Treatment with TTO at 50 µL/L was able to extend longevity of infected fish, and showed 88% of therapeutic success, even though it did not show curative efficacy. TTO treatment was not toxic under these tested concentrations, since biomarkers of hepatic and renal functions were not affected, and the concentration of 50 µL/L was able to prevent increased levels of aspartate aminotransferase. There was no significative differences regarding hematological parameters (p < 0.05). Treatment with TTO 50 µL/L was able to reduce histopathological alterations usually caused by this type of bacteria in the gills, but it was unable to reduce hepatic histopathological alterations. Our results showed, for the first time, that TTO has high activity against A. hydrophila and proved to be a natural alternative to prevent and control this pathogen.

Influence of Melaleuca alternifolia oil nanoparticles on aspects of Pseudomonas aeruginosa biofilm.

The Pseudomonas aeruginosa is a gram-negative bacillus and frequent cause of infection. This microorganism is resistant intrinsically to various drugs. The P. aeruginosa is associated with the biofilm formation, which causes worsen the prognosis and difficulty the treatment. The influence of Melaleuca alternifolia oil or "tree of tee" oil (TTO) and TTO nanoparticles on adhesion of P. aeruginosa in buccal epithelial cells was investigated. Also was determined the antimicrobial and antibiofilm activity against this microorganism. The TTO nanoparticles were produced by deposition of preformed polymer and the physic-chemical properties of nanoparticles were measured by electrophoresis and dynamic light scattering. The characterization of nanoparticle showed acceptable values for diameter and zeta potential. The evaluation of antimicrobial and antibiofilm activity against P. aeruginosa PAO1 was performed by microdilution indicating the minimal inhibitory concentration, and the potential antibiofilm. It was verified the action on virulence factors such the motility, besides the influence on adhesion in buccal epithelial cells. Both oil and nanoparticles showed a decrease in adhesion of microorganisms to buccal cells, decrease of biofilm and interfering on P. aeruginosa PAO1 motility. The nanostructuration of TTO, shows be a viable alternative against formed biofilm microorganisms.

Co-encapsulation of imiquimod and copaiba oil in novel nanostructured systems: promising formulations against skin carcinoma.

In this study, two types of cutaneous-directed nanoparticles are proposed for the co-encapsulation of imiquimod (a drug approved for the treatment of basal cell carcinoma) and copaiba oil (oil that exhibits anti-proliferative properties). Nanostructured copaiba capsules (NCCImq) were prepared using the interfacial deposition method, and nanostructured Brazilian lipids (NBLImq) were prepared by high-pressure homogenization. The formulations exhibited average diameter, zeta potential, pH and drug content of approximately 200nm, -12mV, 6 and 1mgmL(-1), respectively. In addition, the formulations exhibited homogeneity regarding particle size, high encapsulation efficiency and stability. Both nanocarriers controlled imiquimod release, and NBLImq exhibited slower drug release (p < 0.05), likely due to increased interaction of the drug with the solid lipid (cupuaçu seed butter). The in vitro evaluation of the imiquimod-loaded nanocarriers was performed using healthy skin cells (keratinocytes, HaCaT); no alteration was observed, suggesting the biocompatibility of the nanocarriers. In addition, in vitro skin permeation/penetration using pig skin was performed, and NCCImq led to increased drug retention in the skin layers and reduced amounts of drug found in the receiver solution. Thus, NCCImq is considered the most promising nanoformulation for the treatment of skin carcinoma.

Effect of tea tree oil (Melaleuca alternifolia) on the longevity and immune response of rats infected by Trypanosoma evansi.

This study aimed to evaluate the effect of tea tree oil (TTO - Melaleuca alternifolia) on hepatic and renal functions, and the immune response of rats infected by Trypanosoma evansi. A pilot study has shown that rats treated with TTO orally (1 ml kg(-1)) had increased survival rate without curative effect. In order to verify if increased longevity was related to a better immune response against T. evansi when using tea tree oil, a second experiment was conducted. Thus, twenty-four rats were divided into four groups. The groups A and B were composed of uninfected animals, and the groups C and D had rats experimentally infected by T. evansi. Animals from the groups B and D were treated orally with TTO (1 ml kg(-1)) for three days. Blood samples were collected to verify humoral response analysis for immunoglobulins (IgA, IgM, IgE, and IgG) and cytokines (TNF-α, INF-γ, IL-1, IL-6, IL-4, and IL-10) at days 0, 3, 5 and 15 post-infection (PI). TTO treatment caused changes in the immunoglobulins and cytokines profile, as well as the course of T. evansi infection in rats. It was found that the TTO was not toxic, i.e., hepatic and renal functions were not affected. Therefore, it is possible to conclude that TTO influences the levels of inflammatory mediators and has trypanocidal effect, increasing life expectancy of rats infected by T. evansi.

Trypanocidal action of tea tree oil (Melaleuca alternifolia) against Trypanosoma evansi in vitro and in vivo used mice as experimental model.

This study aimed to evaluate the Trypanosoma evansi susceptibility to tea tree oil (TTO - Melaleuca alternifolia) and tea tree oil nanocapsules (TTO nanocapsules) in vitro and in vivo tests. In vitro, we observed a mortality curve of trypomastigotes proportional to dose, i.e., the TTO and TTO nanocapsules have trypanocidal effect. Treatment with TTO in vivo was assessed in experiments (I and II). For Experiment I, T. evansi infected mice were treated with TTO and/or combinations of essential oil with chemotherapy (diminazene aceturate - D.A.). Treatment with TTO at a dose of 1mLkg(-1) was able to extend animal longevity, but had no curative efficacy. However, when TTO was combined with D.A. a disease curative efficacy of 100% for disease was observed, a much better result than the D.A. treatment (33.3%). In Experiment II, T. evansi infected mice were treated with TTO nanocapsules with doses of 0.3, 0.6 and 0.9mLkg(-1). Animals treated with 0.9mLkg(-1) showed higher longevity however without curative effect. Active compounds present in natural products, such as M. alternifolia, may potentiate the treatment of trypanosomosis when associated with other trypanocidal drugs.