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.

Assessment of the biological potential of diaryltriazene-derived triazene compounds.

In the present study, novel, 1,3-diaryltriazene-derived triazene compounds were synthesized and tested. Triazenes are versatile and belong to a group of alkylating agents with interesting physicochemical properties and proven biological activities. This study describes the synthesis, molecular and crystalline structure, biological activity evaluation, and antifungal and antimicrobial potentials of 1,3-bis(X-methoxy-Y-nitrophenyl)triazenes [X = 2 and 5; Y = 4 and 5]. The antimicrobial and antifungal activities of the compounds were tested by evaluating the sensitivity of bacteria (American Type Culture Collection, ATCC) and clinical isolates to their solutions using standardized microbiological assays, cytotoxicity evaluation, and ecotoxicity tests. The antimicrobial potentials of triazenes were determined according to their minimum inhibitory concentrations (MICs); these compounds were active against gram-positive and gram-negative bacteria, with low MIC values. The most surprising result was obtained for T3 having the effective MIC of 9.937 µg/mL and antifungal activity against Candida albicans ATCC 90028, C. parapsilosis ATCC 22019, and C. tropicallis IC. To the best of our knowledge, this study is the first to report promising activities of triazene compounds against yeast and filamentous fungi. The results showed the potential utility of triazenes as agents affecting selected resistant bacterial and fungal strains.

Anti-Ehrlichia properties of the dichloromethane fraction of Ageratum conyzoides associated with doxycycline: In vitro study / Propriedades anti-Ehrlichia da fração diclorometânica de Ageratum conyzoides associada com doxiciclina: estudo in vitro

ABSTRACT: The increasing number of cases of canine ehrlichiosis caused by Ehrlichia canis in hospitals and veterinary clinics has demonstrated the need for a new drug protocol for this disease. Doxycycline is used to treat ehrlichiosis, but the resistance of the microorganism to this treatment protocol, as well as the various side effects to the animals, has become a concern. Several studies have shown a positive interaction with extracts of plants and drugs, which allow for the reduction of the concentration necessary to produce the desired effect, minimizing adverse effects. This study determined the efficiency of the combination of the dichloromethane (DCM) fraction of Ageratum conyzoides L. with anti-Ehrlichia activity and doxycycline by using the checkerboard assay. Plant material was collected in São Luís, northeastern Brazil, followed by extraction in MeOH: H2O (8:2) and partitioning of the DCM fraction. After determining the Minimum Inhibitory Concentration (MIC) of the fraction under study against DH82 cells infected with Ehrlichia canis, it was combined with doxycycline to derive the Fractional Inhibitory Concentration Index (CIF Index). A reduction of 5.83 times the doxycycline minimum inhibitory concentration was observed, showing that this fraction of A. conyzoides composed predominantly by the class of lignans, identified by mass spectrometry notably intensified the activity of doxycycline against E. canis, resulting in a synergistic effect.

RESUMO: O crescente número de casos de erliquiose canina por Ehrlichia canis em hospitais e clínicas veterinárias tem demonstrado a necessidade de um novo protocolo de medicamentos para essa doença. A doxiciclina é usada para tratar a erliquiose, mas a resistência do microrganismo a esse protocolo de tratamento, bem como os diversos efeitos colaterais para os animais, tornou-se uma preocupação. Vários estudos têm demonstrado interação positiva com extratos de plantas e fármacos, que permitem a redução da concentração necessária para produzir o efeito desejado, minimizando os efeitos adversos. Este estudo determinou a eficiência da combinação da fração diclorometânica (DCM) de Ageratum conyzoides L. com atividade anti-Ehrlichia canis associada com doxiciclina por meio do ensaio de Checkerboard. O material vegetal foi coletado em São Luís, Maranhão, nordeste do Brasil, seguido pela extração em MeOH:H2O (8:2) e partição da fração diclorometânica. Após a determinação da Concentração Inibitória Mínima (CIM) da fração em estudo frente às células DH82 infectadas com Ehrlichia canis, a mesma foi combinada com a doxiciclina para derivação do Índice de Concentração Inibitória da Fração (Índice CIF). Observou-se uma redução de 5,83 vezes a concentração inibitória mínima da doxiciclina mostrando que esta fração de A. conyzoides, composta predominantemente por lignanas identificadas por espectrometria de massas, notavelmente intensificou a atividade desse fármaco contra E. canis, resultando em um efeito sinérgico.

Essential oils and their chemical constituents against Aedes aegypti L. (Diptera: Culicidae) larvae.

This review focused on the toxicity of essential oils and their constituents against Aedes aegypti L. (Diptera, Culicidae) larvae, a key vector of important arboviral diseases, such as dengue, chikungunya, zika, and yellow fever. This review is based on original articles obtained by searching major databases in the last six years. Our literature review shows that 337 essential oils from 225 plant species have been tested for larvicidal bioactivity. More than 60% of these essential oils were considered active (LC50<100 µg/mL). Most species belong to the families Lamiaceae (19.3%), Lauraceae (9.9%), and Myrtaceae (9.4%). The plants studied for their larvicidal activity against A. aegypti were mainly collected in India and Brazil (30 and 20%, respectively) and the parts of the plants most used were the leaves. Less than 10% of essential oils were evaluated for toxicity against non-target organisms and with the aim to demonstrate safe use. The most used plant parts are leaves and the main compounds of essential oils were described. The most active essential oils are rich in sesquiterpene hydrocarbons, oxygenated sesquiterpenes, and monoterpene hydrocarbons. Here, factors affecting bioactivity (chemical composition, plant parts, and harvesting site) of essential oils and their constituents, as well as safety to non-target organisms are discussed. Essential oils have considerable potential against A. aegypti. This review shows that essential oils might be used to control arboviruses, and further studies on safety and formulations for application in the field should be performed.

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.

Molluscicidal effect of Euphorbia umbellata (Pax) Bruyns latex on Biomphalaria glabrata, Schistosoma mansoni host snail.

Euphorbia umbellata (Pax) Bruyns is an easily cultivated shrub, with occurrence in the tropical regions of the American and African continents. Chemical studies have revealed that the latex of this plant is rich in terpene compounds, which are highly toxic to snails Biomphalaria glabrata (Basommatophora: Planorbidae). The aim of this study was to evaluate the chemical composition and molluscicidal activity of the latex produced by E. umbellata, as well as the safety of its application in aquatic environments. The concentration of latex that killed 90% of the exposed snails after 24 h exposure (LC90) was 3.69 mg/L. Toxicity bioassays using Danio rerio (zebrafish) revealed that these animals were less susceptible to latex than planorbids. However, it is important to perform other toxicity tests to ensure the feasibility of using latex to control populations of mollusks that contribute to schistosomiasis transmission. A phytochemical screening performed with the E. umbellata latex identified the triterpenoid and coumarin class. Further studies are warranted to isolate, identify, and test the active compounds of E. umbellata latex in B. glabrata.