Seasonal variation in the chemical composition and larvicidal activity against Aedes aegypti L. of essential oils from Brazilian Amazon.

Arthropod-borne viral diseases are important public health threats in tropical countries. In the Americas, diseases such as Dengue, Zika, Chikungunya, Mayaro, and Yellow fever, which are mainly transmitted by Aedes aegypti L. are responsible for high rates of morbidity and mortality in the populations. The incidence of those viruses has increased in the last few years, due to the decrease of vector control programs, as well as, the resistance of this vector in front of many products that have been used in this proposal. Plant-based products such as essential oils are a promising alternative for Ae. aegypti control. In this context, the present work aims to investigate the seasonality effect on the larvicidal potential and safety of essential oils from 4 endemic plants of the Brazilian Amazon, as well as to verify their larvicidal activity and safety. The species were collected in the rainy and dry periods (Eugenia uniflora L., Lantana camara L., Ocimum basilicum L., Plectranthus neochilus Schlrt.). The essential oils were extracted by hydrodistillation and chemical composition was determined by gas chromatography coupled to mass spectrometry. The larvicidal activity was performed according to the methodology recommended by the World Health Organization. The safety of the oil use was evaluated against two non-target organisms: Artemia salina Leach and Danio rerio Hamilton Buchanan. Essential oils of E. uniflora, L. camara, O. basilicum, P. neochilus, presented as major compounds, in both collection periods, the substances Curzerene, Germacrene D, Methyl chavicol and, Caryophyllene, respectively. However, showed a qualitative and quantitative variation of this composition. All oils showed promising results against Ae. aegypti larvae with LC50 between 41.7 and 275.8 µg/mL. They were efficient against Ae. aegypti but showed high toxicity to non-target organisms, requiring additional safety studies. Despite these results showing that those essential oils are not ideal larvicides because they presented toxicity to non-target organisms, bioprospection is a promising tool to help reduce the dissemination of arboviruses, and they can use in artificial breeders of Ae. aegypti larvae, where water will not be reused for human consumption or have other cohabiting organisms.

Molluscicidal and cercaricidal activities of the essential oil of Dysphania ambrosioides (L.) Mosyakin & Clemants: Implications for the control of schistosomiasis.

Schistosomiasis is one of the most important tropical diseases. A fundamental strategy to control its spread is the use of natural products against its vectors, which are snails of the genus Biomphalaria. The present study evaluated the chemical composition, the molluscicidal and cercaricidal effects, and the ecotoxicity of the essential oil from the aerial parts of Dysphania ambrosioides (L.) Mosyakin & Clemants (DAEO). The essential oil was obtained by hydrodistillation and analyzed by gas chromatography-mass spectrometry (GC-MS). Molluscicidal and cercaricidal activities were determined by the immersion method. Environmental toxicity was assessed from bioassays using Artemia salina larvae and Danio rerio fish. DAEO presented a 0.8% yield. The GC-MS analysis revealed the predominance of hydrocarbon monoterpenes in the oil. A total of 32 constituents was identified, with α-terpinene (50.69%) being the major compound, followed by p-cymene (13.27%) and ascaridole (10.26%). DAEO was active against adult Biomphalaria glabrata snails and demonstrated lethal effect against Schistosoma mansoni cercariae, with LC50 values of 25.2 (22.7-27.8) and 62.4 (61.8-62.9) µg/mL, respectively. Regarding toxicity to non-target aquatic organisms, the oil showed LC50 values of 86.9 (84.7-87.6) and 18.6 µg/mL (15.5-22.8) for A. salina and D. rerio, respectively. DAEO proved to be a promising natural product for the control of schistosomiasis, acting on both the vectors and the etiological agent of the disease. However, the use of the oil is safer in transmission sites where there are no non-target organisms, as it has showed toxicity to D. rerio fish.

Molluscicidal activity of monoterpenes and their effects on inhibition of acetylcholinesterase activity on Biomphalaria glabrata, an intermediate host of Schistosoma mansoni.

The molluscicidal action of essential oils have been attributed to the most prevalent terpene compounds. However, molluscicidal properties, mode of action, and toxicity to non-target organisms remain unclear. In this study, the molluscicidal potential of four monoterpenes (camphor, thymol, α-pinene, and 1,8-cineole) against the snail Biomphalaria glabrata, an intermediate host of Schistosoma mansoni, was analyzed. The molluscicide activity of each monoterpene was assessed by the standardized test of the World Health Organization (WHO) and the monoterpenes considered active against B. glabrata were analyzed as inhibitors of the enzymatic activity of acetylcholinesterase (AChE) extracted from snails. In addition, acute toxicity to non-target organisms was assessed against Danio rerio fish. The results show that camphor and 1,8-cineole monoterpenes did not induce snail mortality. Thymol and α-pinene were active against B. glabrata, inducing mortality in concentration-dependent patterns and showing a lethal effect in concentrations compatible with that recommended by the WHO (LC90 of 7.11 and LC90 10.34 µg ∙ mL-1, respectively). The toxic action of thymol and α-pinene on snails indicates that these monoterpenes may account for or largely contribute to the molluscicidal activity of essential oils that contain them as major compounds. Thymol and α-pinene inhibit the AChE of B. glabrata at concentrations higher than those used in the molluscicide test. These monoterpenes show low toxicity to non-target organisms compared to the commercial molluscicide niclosamide. Knowledge about monoterpene toxicity against B. glabrata contributes to its potential use in molluscicidal formulations and in alternatives to the control of snails that host intermediate S. mansoni, a crucial action in the prevention and transmission of schistosomiasis, a neglected tropical disease.

Essential oils as molluscicidal agents against schistosomiasis transmitting snails - a review.

This review aims to describe essential oils with bioactivity on adult snails of the genera Bulinus and Biomphalaria, which are intermediate hosts of schistosomes, and brings together information relating to the importance of molluscicides and the chemical composition and toxicity of such oils for other aquatic species. Analysis of the data of original articles revealed that 50 essential oils obtained from 46 plant species were evaluated for molluscicidal activity against the genera cited. More than 80% of the volatile oils studied were active, according to the criteria of the World Health Organization (LC90 or LC100 ≤100 µg/mL or LC50 <40 µg/mL), and most of the oils came from plants belonging to the Rutaceae, Lamiaceae and Pinaceae. Around 37% of the surveyed plant species were obtained in Brazil and 88% of these plants were collected in the northeast of the country, a region with a high prevalence of schistosomiasis mansoni. The essential oils with the highest toxicity against host snails had high levels of hydrocarbon monoterpenes and oxygenated monoterpenes, which may be responsible for the molluscicidal activity. Some volatile components were subjected to molluscicidal evaluation, and the monoterpene compounds exhibited a significant molluscicide effect. This review confirmed the importance of essential oils as a promising alternative for the development of natural molluscicide products. However, in order to be safe for use at sites where schistosome intermediate hosts are found, information on ecotoxicity is required and, to date, few oils have been tested against non-target aquatic species.

Anti-Ehrlichia properties of the essential oil of Ageratum conyzoides L. and its interaction with doxycycline.

Canine Monocytic Ehrlichiosis (CME) is an infectious disease caused by the rickettsia organism Ehrlichia canis which is transmitted mainly the ixodid brown dog tick Rhipicephalus sanguineus. The prevalence of E. canis infection has been increasing in recent years. The World Health Organization has been warned about antibiotics resistance and one of the way to prevent this situation is found new compound with this property. Doxycycline is the treatment of choice for this tick-borne disease. Adverse effects are noted in dogs that are sensitive to this drug. Antibiotic resistance may also occur. The present study aimed to evaluate the anti-Ehrlichia properties of the essential oil of the aerial parts of Ageratum conyzoides L. in infected DH82 cells, as well as its anti-Ehrlichia activity associated with doxycycline using the checkerboard assay. A. conyzoides is a native plant from northeast Brazil with many reports of ethnopharmacological applications. The essential oil of A. conyzoides was extracted from the aerial parts of the plant using the hydrodistillation method. E. canis-infected DH82 cells were cultured in DMEM (Dulbecco's Modified Eagle Medium), maintained at 37 °C and 5% CO2, and standardized at a 70% infection rate for the initiation of treatment protocols. The tests were first carried out with the aim of defining the IC50. The combined effect of doxycycline and A. conyzoides essential oil was then determined using the checkerboard dilution technique (checkerboard method) in which the IC50 was 200 µg/mL. The doxycycline reduction index from the combined effect was 4.90 times resulting in a synergistic effect. To the authors' knowledge, this is the first alternative treatment (alternative therapy) based on bioactive molecules that have antibacterial activity against E. canis.