Molecular studies with Aedes (Stegomyia) aegypti (Linnaeus, 1762), mosquito transmitting the dengue virus.

Dengue is an infectious viral disease, which can present a wide clinical picture, ranging from oligo or asymptomatic forms, to bleeding and shock, and can progress to death. The disease problem has increased in recent years, especially in urban and suburban areas of tropical and subtropical regions. There are five dengue viruses, called serotypes (DEN-1, DEN-2, DEN-3, DEN-4, and DEN-5), which belong to the Flaviviridae family and are transmitted to humans through infected mosquito bites, with the main vector the Aedes aegypti mosquito (Linnaeus, 1762). Studies performed with Ae. aegypti, aimed at their identification and analysis of their population structure, are fundamental to improve understanding of the epidemiology of dengue, as well for the definition of strategic actions that reduce the transmission of this disease. Therefore, considering the importance of such research to the development of programs to combat dengue, the present review considers the techniques used for the molecular identification, and evaluation of the genetic variability of Ae. aegypti.

Chemical Composition and Larvicidal Activity of Essential Oils Extracted from Brazilian Legal Amazon Plants against Aedes aegypti L. (Diptera: Culicidae).

The mosquito Aedes aegypti L. (Diptera: Culicidae) is the major vector of dengue and chikungunya fever. The lack of effective therapies and vaccines for these diseases highlights the need for alternative strategies to control the spread of virus. Therefore, this study investigated the larvicidal potential of essential oils from common plant species obtained from the Chapada das Mesas National Park, Brazil, against third instar A. aegypti larvae. The chemical composition of these oils was determined by gas chromatography coupled to mass spectrometry. The essential oils of Eugenia piauhiensis Vellaff., Myrcia erythroxylon O. Berg, Psidium myrsinites DC., and Siparuna camporum (Tul.) A. DC. were observed to be mainly composed of sesquiterpene hydrocarbons. The essential oil of Lippia gracilis Schauer was composed of oxygenated monoterpenes. Four of the five tested oils were effective against the A. aegypti larvae, with the lethal concentration (LC50) ranging from 230 to 292 mg/L after 24 h of exposure. Overall, this work demonstrated the possibility of developing larvicidal products against A. aegypti by using essential oils from the flora of the Brazilian Legal Amazon. This in turn demonstrates the potential of using natural resources for the control of disease vectors.

Syzygium cumini (L.) Skeels essential oil and its major constituent α-pinene exhibit anti-Leishmania activity through immunomodulation in vitro.

ETHNOPHARMACOLOGICAL RELEVANCE: Syzygium cumini (L.) Skeels (Myrtaceae), commonly known as "jambolão" in Brazil is widely used in folk medicine against leishmaniasis, inflammation, chronic diarrhea, and ulcers. It is one of the most commonly used plants for the treatment of diabetes worldwide. In previous studies, Syzygium cumini was shown to possess antihyperlipidemic and anti-allergic properties, and to exhibit good performance as an antimicrobial agent against bacteria, fungi, and protozoa parasites of the genus Leishmania and Trypanosoma. This study was aimed at evaluating the effects of S. cumini essential oil (ScEO) and its major component α-pinene on Leishmania (Leishmania) amazonensis, as well as their cytotoxicity and possible mechanisms of action. MATERIALS AND METHODS: To evaluate the anti-proliferative effect on Leishmania, effects on promastigote and axenic amastigote forms were assessed using tetrazolium salt (MTT) assay. The intramacrophagic amastigotes were exposed to ScEO and α-pinene to determine the survival index. To gain insight into the mechanism of action involved in the effect on the samples, we evaluated the modulation of macrophage activation state by observing structural (phagocytic and lysosomal activities) and cellular (nitric oxide increase) changes. To assess the safety profile of ScEO and α-pinene, murine macrophages and human red blood cells were treated with ScEO and α-pinene and the selectivity index was calculated for each treatment. RESULTS: α-Pinene was effective against Leishmania amazonensis promastigote forms, with a half-maximal inhibitory concentration (IC50) value of 19.7µg/mL. α-Pinene was more active (IC50 values of 16.1 and 15.6µg/mL against axenic and intracellular amastigotes, respectively) than ScEO (IC50 values of 43.9 and 38.1µg/mL against axenic and intracellular amastigotes, respectively). Our results showed that the anti-Leishmania effects were mediated by immunomodulatory activity, as evidenced by the observed increases in both phagocytic and lysosomal activity, and the elevated NO levels. ScEO and α-pinene exhibited low cytotoxicity against murine macrophages and human erythrocytes. The 50% cytotoxicity concentration (CC50) values for the macrophages in the MTT assay were 614.1 and 425.2µg/mL for ScEO and α-pinene, respectively, while the corresponding half-maximal hemolytic concentration (HC50) values were 874.3 and 233.3µg/mL. CONCLUSIONS: Taken together, the results demonstrate that ScEO and its major constituent α-pinene have significant anti-Leishmania activity, modulated by macrophage activation, with acceptable levels of cytotoxicity in murine macrophages and human erythrocytes. Further work is warranted, involving more in-depth mechanistic studies and in vivo investigations.

Essential oils and their compounds as Aedes aegypti L. (Diptera: Culicidae) larvicides: review.

This review aims to describe essential oils and their constituent compounds that exhibit bioactivity against Aedes aegypti L. (Diptera: Culicidae) larvae, the immature stage of the primary vector of dengue. This review is based on original articles obtained by searching on major databases. Our literature review revealed that 361 essential oils from 269 plant species have been tested for their larvicidal activity. More than 60 % of these essential oils were considered active (LC50<100 mg/L), and the majority of these active oils were derived from species belonging to Myrtaceae, Lamiaceae, and Rutaceae. The most active essential oils exhibited effective concentrations comparable with the dosage recommended for the use of temephos in container breeding. Approximately 27 % of the plants studied for their larvicidal activity against A. aegypti were collected in Brazil. Essential oils rich in phenylpropanoids, oxygenated sesquiterpenes, and monoterpene hydrocarbons were found to be the most active. When the isolates were tested, phenylpropanoids and monoterpene hydrocarbons were the most active compound classes. We describe the plant parts used and the major constituents of the essential oils. In addition, we discuss factors affecting the activity (such as plant parts, age of the plant, chemotypes, larval source, and methods used), structure-activity relationships, and mechanisms of action of the essential oils and their compounds. Essential oils have been widely investigated and show high larvicidal activity against A. aegypti. This review reveals that the essential oils are effective alternatives for the production of larvicides, which can be used in vector-borne disease control programmes.

Eugenia uniflora L. Essential Oil as a Potential Anti-Leishmania Agent: Effects on Leishmania amazonensis and Possible Mechanisms of Action.

Eugenia uniflora L. is a member of the Myrtaceae family and is commonly known as Brazilian cherry tree. In this study, we evaluated the chemical composition of Eugenia uniflora L. essential oil (EuEO) by using gas chromatography-mass spectrometry (GC-MS) and assessed its anti-Leishmania activity. We also explored the potential mechanisms of action and cytotoxicity of EuEO. Thirty-two compounds were identified, which constituted 92.65% of the total oil composition. The most abundant components were sesquiterpenes (91.92%), with curzerene (47.3%), γ -elemene (14.25%), and trans- ß -elemenone (10.4%) being the major constituents. The bioactivity shown by EuEO against promastigotes (IC50, 3.04 µ g·mL(-1)) and amastigotes (IC50, 1.92 µ g·mL(-1)) suggested significant anti-Leishmania activity. In the cytotoxicity determination, EuEO was 20 times more toxic to amastigotes than to macrophages. Hemolytic activity was 63.22% at the highest concentration tested (400 µ g·mL(-1)); however, there appeared to be no toxicity at 50 µ g·mL(-1). While the data show that EuEO activity is not mediated by nitric oxide production, they do suggest that macrophage activation may be involved in EuEO anti-Leishmania activity, as evidenced by increases in both the phagocytic capacity and the lysosomal activity. More studies are needed to determine in vivo activity as well as additional mechanisms of the anti-Leishmania activity.