Larvicidal and Adulticidal Activity of Essential Oils from Four Cuban Plants against Three Mosquito Vector Species

Mosquito-borne diseases contribute significantly to disease burden, death, poverty, and social debility all over the world. Among these diseases, dengue, malaria and filariasis are most important diseases spread by mosquitoes. In the present work, the methanolic extract of the unripe fruit peel of Persea americana was tested for the larvicidal activity against the third instar larvae of the three mosquito vectors- Aedes aegypti, Culex quinquefasciatus and Anopheles stephensi for 24 h. The methanol extract of peels showed good larvicidal activity against Anopheles stephensi with LC50 and LC90 values of 6.65 and 71.62 ppm, Aedes aegypti with 7.12 and 86.59 ppm followed by Culex quinquefasciatus with 10.78 and 69.39 ppm respectively. The present study revealed that the methanolic extract of unripe fruit peel of Persea americana showed mosquito larvicidal activity in the order Anopheles stephensi followed by Aedes aegypti and Culex quinquefasciatus in the third instar larval stage.

Swift fabrication of Ag nanostructures using a colloidal solution of Holostemma ada-kodien (Apocynaceae) – Antibiofilm potential, insecticidal activity against mosquitoes and non-target impact on water bugs

Thirty-three species of mosquitoes have been reported from the Kingdom of Saudi Arabia. Several of these mosquitoes, Anopheles gambiae Giles s.l., Anopheles stephensi Liston, Culex pipiens Linnaeus, Culex quinquefasciatus Say, Culex tritaeniorhynchus Giles, Stegomyia aegypti (Linnaeus) and Aedimorphus vexans arabiensis (Patton) are known vectors of human and animal diseases. In this study, the cuticular hydrocarbon profiles of eight mosquito species using gas chromatography-mass spectrometry were analyzed. Wild collected fourth-instar larvae were reared, and single, newly emerged, unfed adult females were used for the analysis. A total of 146-160 peaks were detected from the cuticular extracts by gas chromatography. Repeated analysis of variance (ANOVA) and Tukey HSD Post Hoc test was used to test for quantitative differences in relative hydrocarbon quantity. In addition, a linear regression model was applied using Enter method to determine the diagnostic peaks for the eight mosquito specimens. The ANOVA test indicated that relative peaks were significant (P < 0.05) when selected pairs of peaks were compared. Also, seven compounds showed qualitative differences among the five mosquito vectors tested. The classes of constituents present were n-alkanes, monomethylalkanes, dimethylalkanes, trimethylalkanes, alkenes, branched aromatic hydrocarbons, aldehydes and esters. These compounds have a carbon chain length ranging from 8 to 18 carbons. The most abundant compound in all adult mosquito specimens was n-hexylacrylate [retention time (RT) 6.73 min], which was not detected in Cx. pipiens. In Cx. pipiens, the most abundant peak was benzaldehyde (RT 2.98 min). Gas chromatography-mass spectrometry is a suitable method to identify adult mosquitoes, especially from focal areas of public health concern such as Jazan Province, Saudi Arabia. This method allows a wide range of adult collected material to be identified with high accuracy.

The eco-friendly control of mosquito vectors is a crucial challenge of public health importance. Here we evaluated the larvicidal potential of Artemisia absinthium essential oil (EO) and its three major chemical constituents against six mosquito vectors: Anopheles stephensi, Anopheles subpictus, Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and Culex tritaeniorhynchus. The EO was obtained by leaf hydro-distillation. Its chemical composition was analyzed using gas chromatography-mass spectrometry. Major components were (E)-β-farnesene (31.6%), (Z)-en-yn-dicycloether (11.12%), and (Z)-β-ocimene (27.8%). The EO was toxic effect against larval populations of An. stephensi, An. subpictus, Ae. aegypti, Ae. albopictus, Cx. quinquefasciatus, and Cx. tritaeniorhynchus, with LC50 values of 41.85, 52.02, 46.33, 57.57, 50.57, and 62.16μg/ml. (E)-β-farnesene, (Z)-en-yn-dicycloether, and (Z)-β-ocimene were highly effective on An. stephensi (LC50 = 8.13, 16.24 and 25.84μg/ml) followed by An. subpictus (LC50 = 10.18, 20.99, and 30.86μg/ml), Ae. aegypti (LC50 = 8.83,17.66, and 28.35μg/ml), Ae. albopictus (LC50 = 11.38,23.47, and 33.72μg/ml), Cx. quinquefasciatus (LC50 = 9.66, 19.76, and 31.52μg/ml), and Cx. tritaeniorhynchus (LC50 = 12.51,25.88, and 37.13μg/ml). Notably, the EO and its major compounds were safer to the non-target organisms Chironomous circumdatus, Anisops bouvieri and Gambusia affinis, with LC50 values ranging from 207.22 to 4385μg/ml. Overall, our results highlight that (E)-β-farnesene, (Z)-en-yn-dicycloether, and (Z)-β-ocimene from the A. absinthium EO represent promising eco-friendly larvicides against six key mosquito vectors with moderate toxicity against non-target organisms.

Vector-borne diseases caused by mosquitoes are one of the major economic and health problems in many countries. The Aedes aegypti mosquito is a vector of several diseases in humans like yellow fever and dengue. Vector control methods involving the use of chemical insecticides are becoming less effective due to development of insecticides resistance, biological magnification of toxic substances through the food chain, and adverse effects on environmental quality and non-target organisms including human health. Application of active toxic agents from plant extracts as an alternative mosquito control strategy was available from ancient times. These are nontoxic, easily available at affordable prices, biodegradable, and show broad-spectrum target-specific activities against different species of vector mosquitoes. Today, nanotechnology is a promising research domain which has wide-ranging application vector control programs. The present study investigates the larvicidal potential of solvent leaf extracts of Leucas aspera and synthesized silver nanoparticles using aqueous leaf extract against fourth instar larvae of Aedes aegypti. Larvae were exposed to varying concentrations of plant extracts and synthesized AgNPs for 24 h. The results were recorded from UV-Vis spectra, x-ray diffraction (XRD), Fourier transform infrared (FT-IR), and scanning electron microscopy (SEM), and were used to characterize and support the biosynthesis of silver nanoparticles. The formation of the AgNPs synthesized from the XRD spectrum compared with Bragg reflections can be indexed to the (111) orientations, respectively, confirmed the presence of AgNPs. The FT-IR spectra of AgNPs exhibited prominent peaks at 3,447.77; 2,923.30; and 1,618.66 cm(-1). The spectra showed sharp and strong absorption band at 1,618.66 cm(-1) assigned to the stretching vibration of (NH) C═O group. The band 1,383 developed for C═C and C═N stretching, respectively, and was commonly found in the proteins. SEM analysis of the synthesized AgNPs clearly showed the clustered and irregular shapes, mostly aggregated, and having the size of 25-80 nm. Energy-dispersive x-ray spectroscopy showed the complete chemical composition of the synthesized AgNPs. In larvicidal activity, the results showed that the maximum efficacy was observed in synthesized AgNPs leaf extracts against the fourth instar larvae of A. aegypti (LC50 values of 8.5632, 10.0361, 14.4689, 13.4579, 17.4108, and 27.4936 mg/l) and (LC90 values of 21.5685, 93.03928, 39.6485, 42.2029, 31.3009, and 53.2576 mg/l), respectively. These results suggest that the synthesized AgNPs leaf extracts have a higher larvicidal potential as compared to crude solvent extracts thus making them an effective combination for controlling A. aegypti.

Mosquito-borne infectious diseases are a persistent problem in tropical regions of the world, including Southeast Asia. Vector control has relied principally on synthetic insecticides, but these have detrimental environmental effects and there is an increasing demand for plant-based agents to control insect pests. Invasive weedy plant species may be able to serve as readily available sources of essential oils, some of which may be useful as larvicidal agents for control of mosquito populations. We hypothesize that members of the genus Conyza (Asteraceae) may produce essential oils that may have mosquito larvicidal properties. The essential oils from the aerial parts of Conyza bonariensis, C. canadensis, and C. sumatrensis were obtained by hydrodistillation, analyzed by gas chromatography–mass spectrometry, and screened for mosquito larvicidal activity against Aedes aegypti, Ae. albopictus and Culex quinquefasciatus. The essential oils of C. canadensis and C. sumatrensis, both rich in limonene (41.5% and 25.5%, respectively), showed notable larvicidal activities against Ae. aegypti (24-h LC50 = 9.80 and 21.7 μg/mL, respectively) and Ae. albopictus (24-h LC50 = 18.0 and 19.1 μg/mL, respectively). These two Conyza species may, therefore, serve as sources for alternative, environmentally-benign larvicidal control agents.

Mosquitoes are significant public health pests and act as a vector of several diseases viz ., malaria, filariasis, Japanese encephalitis, dengue fever and chikungunya, which are transmitted by the three genera of mosquitoes, viz ., Anopheles, Culex and Aedes . Mosquito control strategies have depended primarily on the use of synthetic chemical insecticides but resulted in rebounding vectorial capacity, environmental and human health concerns. Plants may be a source of alternative agents to replace the synthetic insecticides for mosquito control. In the present study, the isolated fractions of Murraya koenigii hexane leaf extracts were evaluated for larvicidal activity against vector mosquitoes viz., Aedes aegypti , Anopheles stephensi and Culex quinquefasciatus . Six fractions viz., A, B, C, D, E and F were obtained from the residue of hexane extract by column chromatography. Standard WHO protocol with minor modifications was adopted for the larvicidal bioassay. Larvicidal activity was evaluated against the isolated fractions at concentrations of 25, 50, 75 and 100 ppm. Larval mortality was observed 24 hours post exposure. Amongst the isolated fractions tested, fraction ‘D’ showed 100.0, 97.6 and 99.2% mortality against third instar larvae of Aedes aegypti , Culex quinquefasciatus and Anopheles stephensi at 100 ppm, respectively and LC50 values were 35.06, 27.20 and 42.51 ppm respectively. Further investigations are needed to explore the larvicidal activity of the isolated fraction ‘D’ of hexane leaf extract of this plant and also the active principle(s) responsible for larvicidal activity.

Background: Sukumara Ghrita an Ayurvedic ghee-based formulation is renowned for its efficacy in addressing various ailments. Objective: This study employs advanced analytical techniques, specifically High-Performance Thin-Layer Chromatography (HPTLC) and Gas Chromatography-Mass Spectrometry (GC-MS), to comprehensively characterize Sukumara Ghrita and to explore bioactive principles present in Sukumara Ghrita and understand their probable mode of action in treating various ailments. Materials and Methods: The medicine was procured from GMP certified Sitaram Ayurveda Pharma and was subjected to HPTLC and GC–MS analysis after due processing. The methanolic extracts were used for HPTLC and GCMS studies. HPTLC fingerprints for methanolic extracts of Sukumara Ghrita was established with good separation and resolution in solvent system Toluene: Ethyl acetate: Hexane (6:3:1). Results: GC-MS analysis of Sukumara Ghrita revealed the presence of 11 phytochemical compounds. HPTLC analysis detected the presence of 9 and 14 active biological constituents at 254 nm and 366 nm respectively, which remains to be identified. Conclusion: The probable mode of action of Sukumara ghrita can be understood through the action of phytoconstituents screened through HPTLC and GC-MS. The phytoconstituents screened are said to have Anti-oxidant, Anti-hypercholesterolemic, Antihypertriglyceridemic, Anti-inflammatory etc actions.

Medicinal plants were used to treat diseases traditionally since ancient times. The present work aims to investigate the bioactive constituents through GC MS analysis of ethanolic leaf extracts of A.marina. Phytochemical screening confirmed the presence of Alkaloids, Flavonoids, Phenols, Reducing sugars, Saponins, Tannin, Glycoside, Triterpenoids and Carbohydrate in A.marina. The characterization of the compounds by Gas Chromatography – Mass Spectrometry (GC-MS) technique has reported the presence of thirty compounds in A.marina leaves. These compounds possess different pharmacological properties like anti-microbial, antioxidant, anti-inflammatory and hepatoprotective properties. Keywords: Avicennia marina, GC-MS, Bioactive compounds, Antibacterial activity.

Three organophosphorus compounds- malathion, folithion and temephos- and two synthetic pyrethroids- alphamethrin and deltamethrin- were used for monitoring the susceptibility status of larvae and adults of six vector mosquito species: Culex quinquefasciatus (Filariasis) and Aedes albopictus (Dengue) (both laboratory and field strains); laboratory strains of Aedes aegypti (Dengue), Anopheles stephensi and Anopheles culicifacies (Malaria), and Culex tritaeniorhynchus (Japanese encephalitis) in India. From the LC50 values obtained for these insecticides, it was found that all mosquito species including the field strains of Cx. quinquefasciatus and Ae. albopictus were highly susceptible Except for Cx. quinquefasciatus (field strain) against malathion, 100% mortality was observed at the discriminating dosages recommended by World Health Organization. The residual effect of alphamethrin, deltamethrin, malathion and folithion at 25 mg (ai)/m2 on different surfaces against six species of vector mosquitoes showed that alphamethrin was the most effective on all four treated surfaces (mud, plywood, cement and thatch). Nevertheless, residual efficacy lasted longer on thatch than on the other surfaces. Therefore, synthetic pyrethroids such as alphamethrin can be effectively employed in integrated vector control operations.

Alternaria porri causing purple blotch disease of onion is a destructive phytopathogen which causes severe loss in productivity. The present study aimed to unravel the antagonistic potential and efficacy of volatile organic compounds produced by various Trichoderma spp. against A. porri causing purple blotch disease of onion through Gas chromatography-mass spectrometry (GCMS) analysis. Ten isolates of Trichoderma species were isolated from the rhizospheric soil of healthy onion plants. Upon paired plate technique, the in vitro efficacy of ten Trichoderma isolates were tested against virulent isolate of Alternaria porri isolated from purple blotch disease infected onion plants. The Trichoderma isolate TIM2 showed 76.29 per cent inhibition on mycelial growth of pathogen. The effective Trichoderma isolate was identified as Trichoderma hamatum through the analysis of the rDNA of internal transcribed spacers (ITS) region and it was subjected to GC-MS analysis. The result of GCMS analysis indicated the highest peak area and retention time with major antimicrobial bioactive compounds like Tetradecane, 2,6,10-trimethy (20.327), (1.22) and Dodecane, 2-cyclohexyl (20.079), (2.14), Heptadecane (21.222), (9.50), Octadecane (22.379), (3.58), Eicosane, 9-cyclohexyl (22.578), (1.84), 2-Propenoic acid, pentadecyl ester (23.400), (10.37), 2,6,10,14-tetramethyl (23.567), (10.37), Eicosane (27.311), (2.34), Hexadecanoic acid, methyl ester (27.918), (4.43), n-Hexadecanoic acid (29.156), (3.59) and Tetrapentacontane, 1,54-dibromo (31.906), (3.33). These bioactive compounds identified through GCMS analysis from the crude extracts of Trichoderma hamatum exhibited a stronger antifungal activity against A. porri. Hence the application of T. hamatum for the management of purple blotch disease highly supress growth of the pathogen and reduce the disease incidence.

Extensive use of chemical larvicides to control larvae, has led to resistance in vectors. More efforts have been conducted the use of natural products such as plant essential oils and their new formulations against disease vectors. Nanoformulation techniques are expected to reduce volatility and increase larvicidal efficacy of essential oils. In this study for the first time, a larvicide nanoemulsion from the essential oil of Acroptilon repens was developed and evaluated against Anopheles stephensi larvae under laboratory conditions. Fresh samples of A. repens plant were collected from Urmia, West Azarbaijan Province, Iran. A clevenger type apparatus was used for extracting oil. Components of A. repens essential oil (AEO) were identified by gas chromatography-mass spectrometry (GC-MS). All larvicidal bioassay tests were performed according to the method recommended by the World Health Organization under laboratory condition. Particle size and the morphologies of all prepared nanoformulations determined by DLS and TEM analysis. A total of 111 compounds were identified in plant. The LC50 and LC90 values of AEO calculated as 7 ppm and 35 ppm respectively. AEO was able to kill 100% of the larvae in 4 days. The nanoemulsion of AEO showed a weak effect on the larvar mortality. It may therefore be suggested that this kind of nanoemulsion is not appropriate for the formulation as a larvicide. It is important to screen native plant natural products, search for new materials and prepare new formulations to develop alternative interventions with a long-lasting impact.

Oxycodone is a semisynthetic opioid analgesic largely prescribed for post-operative and chronic pain management. The introduction of a slow release formulation of oxycodone has led to its frequent abuse and to an increase in emergency cases related to oxycodone overdose. Until recently, oxycodone testing has been confined to gas chromatography-mass spectrometry (GC-MS) analysis because the widely used automated opiate immunoassays poorly react to this compound. We investigated the utility of a new oxycodone immunoassay as a screening procedure to eliminate inappropriate GC-MS testing of negative urine specimens. We analyzed 96 urine specimens using GC-MS and two immunoassays, CEDIA((R)) opiates and DRI((R)) oxycodone assays from Microgenics, on a Hitachi 917 analyzer. The GC-MS allowed us to detect codeine, hydrocodone, hydromorphone, morphine, oxycodone, and oxymorphone following enzymatic hydrolysis and derivation by acetylation. The combination of the two immunoassays gave the best performance (98% sensitivity and specificity) when considering a positive result from GC-MS for any of the opiates. Considering positive GC-MS results for oxycodone or oxymorphone only, the oxycodone immunoassay resulted in two false-positives and one false-negative (50 ng/mL cutoff). Using these immunoassays for screening before GC-MS analysis provides a reduced opiate GC-MS workload without compromising quality.

Larvicidal bioassay was carried out in the laboratory to assess the potential of ethanolic extracts of dried fruits of three species of peppercorns: Long pepper, Piper longum L., Black pepper, Piper nigrum, and White pepper, Piper nigrum against the different instars of field-collected Indian strain of dengue fever mosquito (Aedes aegypti L.). The investigations established the larvicidal potential of all the varieties of pepper fruits against Ae. aegypti. Against early fourth instar, the ethanolic extracts of Black and White P. nigrum proved to be 30-40% less toxic than the extracts of P. longum, whereas against third instars, white pepper extracts exhibited 7% more efficacy than that of black pepper and 47% more toxicity than that of long pepper. The results also revealed that the extracts of all the three pepper species were 11-25 times more toxic against the third instar larvae as compared to the early fourth instars. The lethal concentration, 50% (LC(50)) values obtained with ethanolic extracts of P. longum, White P. nigrum and Black P. nigrum against early fourth instar larvae were 0.248, 0.356, and 0.405 ppm, respectively, and the lethal concentration, 90% (LC(90)) values were 0.605, 0.758, and 0.801 ppm, respectively. Whereas against third instar larvae, the LC(50) values recorded with three extracts were 0.022, 0.015, and 0.016 ppm and the LC(90) values recorded were 0.054, 0.034, and 0.046 ppm, respectively. The larvae treated with all the pepper species showed initial abnormal behavior in their motion followed by excitation, convulsions, and paralysis, leading to 100% kill indicating delayed larval toxicity and effects of the extracts on the neuromuscular system. Observations of morphological alterations on treated larvae under light microscopy revealed that most organs, except anal papillae, had a normal structural appearance as that of controls. The structural deformation in the form of shrinkage in the internal membrane exhibited by anal papillae suggests the anal papillae as the probable action sites of the pepper extracts. The potential of peppercorns as new types of larvicides for the control of mosquitoes are explored.

Independent evaluation of a commercial deconvolution reporting software for gas chromatography mass spectrometry analysis of pesticide residues in fruits and vegetables