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Chemical composition and larvicidal activity of Ocimum species essential oils from the south of the Paranaense forest (Argentine) against Aedes aegypti (Diptera: Culicidae) larvae

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Abstract

Aedes aegypti plays a predominant role in the transmission of a variety of diseases, such as yellow fever, dengue fever, malaria, Zika and Chikungunya disease. The reduction of chemicals available for vector control due to the increase in mosquito resistance to insecticides and their negative impact on the human health and the environment have driven the focus of the problem towards the identification and evaluation of sustainable alternatives to synthetic insecticides. The essential oils from the leaves of Ocimum campechianum, Ocimum ovatum, and Ocimum selloi were analyzed by GC/MS and GC/FID and investigated for its larvicidal activity against A. aegypti larvae. In the chromatographic analyses showed that the main compounds were elemicin (22.8%), methyl eugenol (33.4%), and methyl eugenol (46.5%), respectively. The susceptibility tested on essential oils was performed according to the World Health Organization protocol. The highest larvicidal potential was displayed by O. ovatum, followed by O. campechianum and O. selloi, with LC50 values of 47.5, 95.5 and 141.2 ppm, respectively. The rapid and efficient larvicidal activity of these three essential oils led us to consider these results as promising for the search of a potentially effective earth-friendly larvicide against the A. aegypti vector.

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Interest in, and research on botanical insecticides has surged since 2000 according to a recent bibliographic analysis of scientific literature. Thousands of papers have now been published documenting the bioactivities of botanicals such as neem (from Azadirachta indica), various plant essential oils, and innumerable plant extracts to insects and related pests, although the majority of these simply report screening studies and other observations made within the confines of laboratories. In contrast, with few exceptions, little of this knowledge has been translated to practice, i.e., utilized directly by farmers for pest management. What, then, are the prerequisites to realizing the benefits of botanical insecticides, particularly for smallholder farmers in sub-Saharan Africa? I have previously argued that there are a number of plant species widely occurring in Africa − both endemic and introduced − that are suitable for the production of botanical insecticides, and that less attention should be paid to further discovery of bioactive plant species and more attention placed on the development of botanicals from those plants we already know. To put those plant species to work for pest management, we need the following: (i) methods for local propagation and cultivation to ensure a sustainable supply of biomass; (ii) simple methods: for extraction utilizing available resources at minimal cost; (iii) simple methods: (e.g., bioassay) to validate bioactivity and ensure some measure of efficacy; and (iv) field trials and demonstrations to learn timing and application strategies that can optimize efficacy. Appropriate technologies for the utilization of botanical preparations for insect control will differ depending on the scale, from the “do-it-yourself” smallholder farmer, to a cottage- or village-level collective, to private-sector industry.
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Mosquitoes (Diptera: Culicidae) are a key threat for millions of people and animals worldwide, since they act as vectors for devastating pathogens and parasites, including malaria, dengue, Japanese encephalitis, filiariasis and Zika virus. Mosquito young instars are usually targeted using organophosphates, insect growth regulators and microbial agents. Indoor residual spraying and insecticide-treated bed nets are also employed. However, these chemicals have negative effects on human health and the environment and induce resistance in a number of vectors. In this scenario, newer and safer tools have been recently implemented to enhance mosquito control. The concrete potential of screening plant species as sources of metabolites for entomological and parasitological purposes is worthy of attention, as recently elucidated by the Y. Tu's example. Here we investigated the toxicity of Heracleum sprengelianum (Apiaceae) leaf essential oil and its major compounds toward third instar larvae of the malaria vector Anopheles subpictus, the arbovirus vector Aedes albopictus and the Japanese encephalitis vector Culex tritaeniorhynchus. GC-MS analysis showed that EO major components were lavandulyl acetate (17.8%) and bicyclogermacrene (12.9%). The EO was toxic to A. subpictus, A. albopictus, and C. tritaeniorhynchus, with LC50 of 33.4, 37.5 and 40.9µg/ml, respectively. Lavandulyl acetate was more toxic to mosquito larvae if compared to bicyclogermacrene. Their LC50 were 4.17 and 10.3µg/ml for A. subpictus, 4.60 and 11.1µg/ml for A. albopictus, 5.11 and 12.5µg/ml for C. tritaeniorhynchus. Notably, the EO and its major compounds were safer to three non-target mosquito predators, Anisops bouvieri, Diplonychus indicus and Gambusia affinis, with LC50 ranging from 206 to 4219µg/ml. Overall, this study highlights that H. sprengelianum EO is a promising source of eco-friendly larvicides against three important mosquito vectors with moderate toxicity against non-target aquatic organisms.
Article
Chemical composition of essential oils obtained from four species of genus Salvia were analyzed by gas chromatography (GC) and gas chromatography mass spectrometry (GC-MS). Main compounds identified from Salvia species essential oils were as follows: 1,8-cineole (71.7%), -pinene (5.1%), camphor (4.4%) and -pinene (3.8%) in S. apiana; borneol (17.4%), -eudesmol (10.4%), bornyl acetate (5%) and guaiol (4.8%) in S. elegans; bornyl acetate (11.4%), -caryophyllene (6.5%), caryophyllene oxide (13.5%) and spathulenol (7.0%) in S. leucantha; -thujene (25.8%), viridiflorol (20.4%), -thujene (5.7%) and camphor (6.4%) in S. officinalis. In biting deterrent bioassays, essential oil of S. leucantha and S. elegans at 10 µg/cm2 showed activity similar to DEET (97%, N, N-diethyl-meta-toluamide) in both the species of mosquitoes whereas activity of S. officinalis and S. apiana was lower than the other oils or DEET. Pure compounds β-eudesmol and guaiol showed biting deterrent activity similar to DEET at 25 nmol/cm2 whereas activity of 13-epi-manool, caryophyllene oxide, borneol, bornyl acetate and β-caryophyllene was significantly lower than β-eudesmol, guaiol or DEET. All essential oils showed larvicidal activity except S. apiana which was inactive at the highest dose of 125 ppm against both mosquito species. Based on 95% CIs, all the essential oils showed higher toxicity in Anopheles quadrimaculatus than Aedes aegypti. Essential oil of S. leucantha with LC50 value of 6.2 ppm showed highest toxicity in An. quadrimaculatus.
Article
Chemical composition analysis of the essential oil from the leaves and flowers of two accessions of Ocimum selloi Benth, cultivated at the Federal University of Viçosa was carried out. For accession A the major component was identified as estragole and represented 94,95% and 92,54% of the oil from the leaves and flowers respectively. For accession B, the oil from the leaves and flowers was constituted by 65,49% and 66,18% of methyleugenol, respectively. For both accessions several minor constituents were also identified. The phenotypic and chemical differences observed between these two accessions suggest the existence of two chemicaly distinct varieties for Ocimum selloi Benth.
Article
Essential oils were extracted from four West African plant species [Tagetes minuta (Family Compositae), Hyptis suaveolens (Family Labiatae), white basil Ocimum canum (Family Labiatae), and sweet basil O. basilicum (Family Labiatae)] by steam distillation. The oil of the pepper Piper guineense (Family Piperaceae), was extracted from the fruits by hydro distillation and ethanol extraction. Mixed essential oil and total ethanol extract was used. Kaolin powder (clay) was mixed (aromatized) with these different oils. Cowpea weevils were reared on chickpeas and newly emerged males and females were deposited on uninfested seeds. Bioassays, i.e. fumigation with pure essential oils and aromatized kaolin powders, were carried out on adults and eggs. Twenty four hours after fumigation, 99 and 0% adult mortality were observed, respectively, as the result of treatments with Ocimum basilicum and the control. The application of powders aromatized with the same oils to weevil pairs resulted in a complete lack of oviposition, whereas 31, 56 and 76 eggs were laid in the controls after 24, 48 and 72 h, respectively. Application of aromatized powders did not have a significant effect on egg hatching (50 out of 110 with O. canum, 100 out of 115 with O. basilicum and 100 out of 130 in the control sample) but did have a significant impact on adult emergence: 0% for the two treatments compared with 100% in the controls. Our results suggest that plants of the genus Ocimum can be used as an alternative to synthetic insecticides.
Article
We investigated the toxicity of (-)-asarinin, α-asarone, methyleugenol, pellitorine, and pentadecane identified in Asarum heterotropoides root to third instar larvae from insecticide-susceptible Culex pipiens pallens (KS-CP strain), Aedes aegypti, and Ochlerotatus togoi as well as field-collected C. p. pallens (DJ-CP colony), identified by polymerase chain reaction. Results were compared with those of two conventional mosquito larvicides: fenthion and temephos. Pellitorine (LC50, 2.08, 2.33, and 2.38 ppm) was 5.5, 10.8, and 25.6 times, 4.5, 11.6, and 24.7 times, and 6.9, 11.1, and 24.6 times more toxic than (-)-asarinin, α-asarone, and methyleugenol against susceptible C. p. pallens, A. aegypti, and O. togoi larvae, respectively. Pentadecane was least toxic. Overall, all the compounds were less toxic than either fenthion or temephos. However, these compounds did not differ in toxicity against larvae from the two Culex strains, even though the DJ-CP larvae exhibited high levels of resistance to fenthion (resistance ratio (RR), 1179), chlorpyrifos (RR, 1174), fenitrothion (RR, 428), deltamethrin (RR, 316), chlorfenapyr (RR, 225), and α-cypermethrin (RR, 94). This finding indicates that the isolated compounds and the pyrethroid, organophosphorus, and pyrrole insecticides do not share a common mode of action or elicit cross-resistance. A. heterotropoides root-derived materials, particularly (-)-asarinin and pellitorine, merit further study as potential mosquito larvicides for the control of insecticide-resistant mosquito populations in light of global efforts to reduce the level of highly toxic synthetic insecticides in the aquatic environment.
Article
The study deals with the efficacy of Ocimum sanctum essential oil (EO) and its major component, eugenol against the fungi causing biodeterioration of food stuffs during storage. O. sanctum EO and eugenol were found efficacious in checking growth of Aspergillus flavus NKDHV8; and, their minimum inhibitory concentrations (MICs) were recorded as 0.3 and 0.2 microl ml(-1), respectively. The O. sanctum EO and eugenol also inhibited the aflatoxin B1 production completely at 0.2 and 0.1 microl ml(-1), respectively. Both of these were found superior over some prevalent synthetic antifungals and exhibited broad fungitoxic spectrum against 12 commonly occurring fungi. The LD50 value of O. sanctum EO on mice was found to be 4571.43 microl kg(-1) suggesting its non-mammalian toxic nature. The findings of present study reveals the possible exploitation of O. sanctum EO and eugenol as plant based safe preservatives against fungal spoilage of food stuffs during storage.
Article
Mosquito larvicidal activity of Piper longum fruit-derived materials against the fourth-instar larvae of Aedes aegypti was examined. A crude methanol extract of P. longum fruits was found to be active against the larvae, and the hexane fraction of the methanol extract showed a strong larvicidal activity of 100% mortality. The biologically active component of P. longum fruits was characterized as pipernonaline by spectroscopic analyses. The LC(50) value of pipernonaline was 0.25 mg/L. The toxicity of pipernonaline is comparable to that of pirimiphos-methyl as a mosquito larvicide. In tests with available components derived from P. longum, no activity was observed with piperettine, piperine, or piperlongumine.
Article
Ocimum spp. (Lamiaceae) and their essential oils have been traditionally used to kill or repel insects, and also to flavor foods and oral products, in fragrances, in folk medicine and as condiments. In Brazil, Ocimum selloi has been used to treat stomachaches and as an anti-inflammatory remedy. This study was performed to provide data on the chemical composition, acute toxicity, mutagenicity, skin irritant potential and mosquito repellency of Ocimum selloi oil. GC/MS analysis of Ocimum selloi oil revealed that its major constituents were methyl-chavicol or estragole (55.3%), trans-anethole (34.2%), cis-anethole (3.9%) and caryophyllene (2.1%). Ocimum selloi oil given by gavage to adult Swiss Webster mice produced no adverse effects at doses as high as 1250 mg/kg body weight. Deaths and symptoms (e.g. hypoactivity, ataxia and lethargy) were observed at doses > or =1500 mg/kg body weight, being females apparently more susceptible than males. Genotoxicity of Ocimum selloi oil was evaluated in the Salmonella/microsome assay without and with S9 mixture. The oil, tested up to the toxicity limit (500-700 microg/plate), was not mutagenic to tester strains TA97a, TA98 and TA100. None of 30 volunteers of either sex exposed to undiluted Ocimum selloi oil (4-h patch test) showed a positive skin irritant reaction. A field test (six volunteers, each individual his/her own control) was carried out to evaluate mosquito (Anopheles braziliensis) repellency of Ocimum selloi oil diluted in ethanol (10% v/v). The median number of mosquito bites on volunteers' skin-recorded for 30 min after application of Ocimum selloi oil (2, range 0-3) was much lower than that noted after application of the solvent alone (19.5, 3-25) (Wilcoxon test, P<0.01). In conclusion, results showed that Ocimum selloi oil is an effective mosquito repellent that presents a low acute toxicity, poses no mutagenic risk and seems not to be irritating to human skin.
Article
The different ways of describing peak positions on gas chromatograms are reviewed. The retention index is preferred to the theoretical nonane system and the relative retention.The equation given by Kováts for the calculation of the retention index in case of isothermal operation is transformed to a more general form to include also the case of linear temperature programmed operation. This generalized equation gives the same retention index for both ways of operation.
Article
Since the middle ages, essential oils have been widely used for bactericidal, virucidal, fungicidal, antiparasitical, insecticidal, medicinal and cosmetic applications, especially nowadays in pharmaceutical, sanitary, cosmetic, agricultural and food industries. Because of the mode of extraction, mostly by distillation from aromatic plants, they contain a variety of volatile molecules such as terpenes and terpenoids, phenol-derived aromatic components and aliphatic components. In vitro physicochemical assays characterise most of them as antioxidants. However, recent work shows that in eukaryotic cells, essential oils can act as prooxidants affecting inner cell membranes and organelles such as mitochondria. Depending on type and concentration, they exhibit cytotoxic effects on living cells but are usually non-genotoxic. In some cases, changes in intracellular redox potential and mitochondrial dysfunction induced by essential oils can be associated with their capacity to exert antigenotoxic effects. These findings suggest that, at least in part, the encountered beneficial effects of essential oils are due to prooxidant effects on the cellular level.
Morfo-Anatomical and chemical characterization in Ocimum ovatum Benth (LAMIACEAE)
  • F Sanchez Gonzales
  • E O Vargas
  • N Kolb Koslobsky
F. Sanchez Gonzales, E.O. Vargas and N. Kolb Koslobsky, Morfo-Anatomical and chemical characterization in Ocimum ovatum Benth (LAMIACEAE). Boletin de la Sociedad Argentina de Botanica, 44, 163 (2009).