Impact of Argemone mexicana extracts on the cidal, morphological, and behavioral response of dengue vector, Aedes aegypti L. (Diptera: Culicidae).

The larvicidal, behavioral, and morphological response of dengue vector, Aedes aegypti treated with deleterious weed, Argemone mexicana, was explored. The 1,000 ppm extracts of A. mexicana leaf, stem, and roots prepared in five different solvents (petroleum ether, hexane, benzene, acetone, and ethanol) were screened for their larvicidal activity against dengue vector establishing the efficacy of petroleum ether and hexane extracts. Other extracts, unable to give 100% mortality, were considered ineffective and discarded from further study. Larvicidal bioassay conducted with selected extracts confirmed the higher efficacy of hexane extracts exhibiting 1.1- to 1.8-fold more potential than the petroleum ether extracts. The results further revealed 1.6- to 2.4-fold higher efficacy of root extracts than those prepared from the leaves and stem of A. mexicana. The hexane root extract of A. mexicana was found to be the most effective larvicide with LC50 value of 91.331 ppm after 24 h of exposure causing 1.8 and 2.4 fold more toxicity as compared to the hexane leaf and stem extracts, respectively. Prolonged exposure of the larvae to the extracts resulted in increased toxicity potential of the extracts. Observations of the treated larvae revealed excitation, violent vertical, and horizontal movements with aggressive anal biting behavior suggesting effect of extracts on their neuromuscular system. Morphological studies of the treated larvae revealed the demelanization of cuticle and shrinkage of internal cuticle of anal papillae indicating the anal papillae as the probable action sites of the A. mexicana extracts. The potential of A. mexicana as new larvicides against dengue vector are being explored.

Larvicidal efficacy of the Citrus limetta peel extracts against Indian strains of Anopheles stephensi Liston and Aedes aegypti L.

The extracts from the peels of Citrus limetta were prepared using hexane and petroleum ether as the solvents. The larvicidal potential of each extract was assessed against dengue fever vector, Aedes aegypti, and malarial vector, Anopheles stephensi, by evaluating the toxicity effects on early fourth instars. Both the extracts were found effective against both the species. The bioassay with hexane extracts resulted in LC(50) values of 132.45 and 96.15 ppm against A. stephensi and A. aegypti, respectively; while the petroleum ether extracts from the C. limetta peels showed LC(50) values of 244.59 and 145.50 ppm, respectively. It revealed that the hexane extracts possessed 1.9-fold more larvicidal potential against A. stephensi and 1.5-fold more efficacy against A. aegypti as compared to the extracts obtained using petroleum ether as solvent. The data further revealed that the extracts were 1.4-1.7 times more effective against A. aegypti as compared to A. stephensi. The qualitative phytochemical study of the extracts showed the presence of terpenoids and flavonoids as the common phytochemical constituents in the extracts suggesting their possible role in the toxicity. Other constituents tested were not detected except alkaloids which were found to be present only in the petroleum ether extract. Further studies are needed to isolate and identify the active principles involved, their mode of action, formulated preparations for enhancing potency and stability, toxicity, and effects on non-target organisms and the environment. This could help in formulating efficient strategies for mosquito control.

Oviposition-altering and ovicidal potentials of five essential oils against female adults of the dengue vector, Aedes aegypti L.

The oviposition deterrence and ovicidal potential of five different essential oils, peppermint oil (Mentha piperita), basil oil (Ocimum basilicum), rosemary oil (Rosemarinus officinalis), citronella oil (Cymbopogon nardus), and celery seed oil (Apium graveolens), were assessed against female adults of the dengue vector, Aedes aegypti L. Multiple concentration tests were carried out where cups containing 1 mL of different concentrations (100%, 10%, 1%, 0.1%) of the oils and 199 mL of water were used for oviposition. The number of eggs laid and the larvae hatched in each cup were scored to evaluate the oviposition deterrent and ovicidal potentials of the oils. Our investigations revealed that the addition of 100% oil (pure oil) caused complete oviposition deterrence except in A. graveolens which resulted in 75% effective repellency. The use of 10% oil resulted in the maximum deterrence of 97.5% as shown by the M. piperita oil while other oils caused 36-97% oviposition deterrence as against the control. The oviposition medium with 1% oil showed decreased deterrent potential with 30-64% effective repellency, the M. piperita oil being exceptional. However, as the concentrations of the oil were reduced further to 0.1%, the least effective oil observed was A. graveolens (25% ER). Also, the M. piperita oil showed much reduced activity (40%) as compared to the control, while the other oils exhibited 51-58% repellency to oviposition. The studies on the ovicidal effects of these oils revealed that the eggs laid in the water with 100% essential oils did not hatch at all, whereas when 10% oils were used, only the R. officinalis oil resulted in 28% egg hatch. At lower concentrations (1%), the oils of M. piperita, O. basilicum, and C. nardus showed complete egg mortality while those of A. graveolens and R. officinalis resulted in 71% and 34% egg hatches, respectively. When used at 0.1%, the O. basilicum oil was found to be the only effective oil with 100% egg mortality, whereas other oils resulted in 16-76% egg mortality, the least mortality caused by the A. graveolens oil. These results suggest that these essential oils can be employed in a resistance-management program against A. aegypti. Further detailed research is needed to identify the active ingredient in the extracts and implement the effective mosquito management program.

Impact of Parthenium hysterophorus leaf extracts on the fecundity, fertility and behavioural response of Aedes aegypti L.

Different extracts of 1,000 ppm were prepared from the leaves of Parthenium hysterophorus using acetone, benzene, petroleum ether, diethyl ether and hexane as the solvents. The efficacy of each extract was assessed against dengue fever vector, Aedes aegypti by evaluating the variations in fecundity, fertility and behavioural response of the female adults. The leaf extracts could cause 70-100% repellency in the oviposition behaviour of the adults. The diethyl ether extract was found to be the most effective extract resulting in maximum effective repellency (99.7%) leading to the highest levels of reduced fecundity and 100% egg mortality followed by benzene extracts causing 93.8% reduced oviposition and 100% ovicidal effect. Hexane and acetone extracts with the least oviposition deterrence of 70-74% and negligible egg mortality (8-9%) proved to be the least effective extracts. The petroleum ether extract had a moderate impact resulting in 93.2% diminished fecundity and 41% ovicidal effect. The behavioural response of female adults of A. aegypti was evaluated by performing spatial repellency and contact irritancy assays. The most significant spatial repellency behaviour was elicited by acetone extracts leading to escape of 80% mosquitoes. Hexane and diethyl ether extracts could cause moderate response with 50-60% escape, while a slight and no reaction was observed on exposure to petroleum ether and benzene extracts, respectively. An interesting observation was the knocked-down activity caused by the hexane extracts with no recovery even after 24 h. A significant contact irritancy response was noticed in the mosquitoes on exposure to acetone leaf extracts resulting in first flight only after 4 s and a total of 12 flights during exposure. No irritancy behaviour was observed on exposure to diethyl ether and benzene leaf extracts. However, as against controls, a slight irritability response was noticed on exposure to hexane leaf extracts resulting in relative irritability of 1.2. Our results suggest the selective efficiency of Parthenium leaf extracts against A. aegypti, as the most effective oviposition deterrent and ovicidal agent was least effective as irritant extract and vice-versa. Further detailed research is needed to identify the active ingredient in the extracts and implement the effective mosquito management programme.

Larvicidal potential of ethanolic extracts of dried fruits of three species of peppercorns against different instars of an indian strain of dengue fever mosquito, Aedes aegypti L. (Diptera: Culicidae).

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.

Impact of the Argemone mexicana Stem Extracts on the Reproductive Fitness and Behavior of Adult Dengue Vector, Aedes aegypti L. (Diptera: Culicidae).

Present investigations evaluated the impact of Argemone mexicana stem extracts on the reproductive fitness of dengue vector, Aedes aegypti, assessed in terms of oviposition deterrent and ovicidal potential. The oviposition deterrent studies of the extracts, prepared using petroleum ether, hexane, benzene, acetone, and ethanol as the solvents, revealed the maximum deterrence potency of the petroleum ether extracts with a significant 15.6% ED at 60 ppm rising by 83.8% at 1000 ppm to 99.4% effective deterrence (ED). Other stem extracts were found to be ineffective at 60 ppm, though resulted in 85.3-96.2% ED and diminished fecundity in A. aegypti at 1000 ppm. Further, A. mexicana stem extracts exhibited moderate ovicidal potential against A. aegypti eggs causing only 42.65-67.85% egg mortality at 1000 ppm, the lowest hatch of 32.15% caused by the benzene extract. Other stem extracts also failed to express effective ovicidal potency with the percent egg hatch ranging between 96.6 and 99.0 at 400 ppm, and 78.8 and 99.0 at 600 ppm. Our results suggest the significant but variable efficacy of A. mexicana stem extracts causing reproductive disadvantage in A. aegypti.

Larvicidal, Repellent, and Irritant Potential of the Seed-Derived Essential oil of Apium graveolens Against Dengue Vector, Aedes aegypti L. (Diptera: Culicidae).

Aedes aegypti L. is one of the primary disease vectors spreading various dreadful diseases throughout the world, specifically over tropics and subtropics. Keeping in view the adverse effects of chemical insecticides-based intervention measures, the eco-friendly and bio-degradable essential oil extracted from the seeds of celery, Apium graveolens were investigated for its efficacy against Ae. aegypti. Larvicidal bioassay carried out with the seed oil against early fourth instars of Ae. aegypti caused an LC50 and LC90 values of 16.10 and 29.08 ppm, respectively, after an exposure to 24 h. The cidal effect of the celery seed oil augmented by 1.2-fold; after an exposure to 48 h; revealing an LC50 value of 13.22 ppm. Interestingly, the seed oil did not cause immediate larval mortality, suggesting a delayed toxicity against the larval stage. Present investigations also revealed remarkable effective repellency of the oil leading to 100% protection till 165 min as compared to control that did not result in any repellency against adult Ae. aegypti. Interestingly, only one bite was recorded in the 165th-min after which only two bites were scored until 180 min of exposure of the adult mosquitoes to the oil. An exciting observation was that the knocked-down effect in adults exposed to 10% oil-impregnated papers. The contact irritancy assays with paper impregnated with 1% celery seed oil caused first flight only after 4 s resulting in an average of 63.66 flights during 15 min of exposure revealing the relative irritability of 26.97. The qualitative phytochemical analysis of the seed oil showed the presence of flavonoids, lactones, and terpenoids as the major constituents suggesting their probable role in the toxicity. Our results confirmed that celery seed essential oil can be used as an efficient larvicide and repellent against Ae. aegypti. The identification of the bioactive components, their mode of action, and studying effects on non-target organisms and the environment would help in devising mosquito-management strategies.

Evaluation of 15 Local Plant Species as Larvicidal Agents Against an Indian Strain of Dengue Fever Mosquito, Aedes aegypti L. (Diptera: Culicidae).

The adverse effects of chemical insecticides-based intervention measures for the control of mosquito vectors have received wide public apprehension because of several problems like insecticide resistance, resurgence of pest species, environmental pollution, toxic hazards to humans, and non-target organisms. These problems have necessitated the need to explore and develop alternative strategies using eco-friendly, environmentally safe, bio-degradable plant products which are non-toxic to non-target organisms too. In view of this, 15 plant species were collected from local areas in New Delhi, India. Different parts of these plants were separated, dried, mechanically grinded, and sieved to get fine powder. The 200 g of each part was soaked in 1000 mL of different solvents separately and the crude extracts, thus formed, were concentrated using a vacuum evaporator at 45°C under low pressure. Each extract was screened to explore its potential as a mosquito larvicidal agent against early fourth instars of dengue vector, Aedes aegypti using WHO protocol. The preliminary screening showed that only 10 plants possessed larvicidal potential as they could result in 100% mortality at 1000 ppm. Further evaluation of the potential larvicidal extracts established the hexane leaf extract of Lantana camara to be most effective extract exhibiting a significant LC(50) value of 30.71 ppm while the Phyllanthus emblica fruit extract was found to be least effective with an LC(50) value of 298.93 ppm. The extracts made from different parts of other five plants; Achyranthes aspera, Zingiber officinalis, Ricinus communis, Trachyspermum ammi, and Cassia occidentalis also possessed significant larvicidal potential with LC(50) values ranging from 55.0 to 74.67 ppm. Other three extracts showed moderate toxicity against A. aegypti larvae. Further investigations would be needed to isolate and identify the primary component responsible for the larvicidal efficiency of the effective plants.

Larvicidal and irritant activities of hexane leaf extracts of Citrus sinensis against dengue vector Aedes aegypti L.

OBJECTIVE: To assess the larvicidal and irritant activities of the hexane extracts of leaves of Citrus sinensis (C. sinensis) against the early fourth instars and female adults of Aedes aegypti (Ae. aegypti). METHODS: The larvicidal potential of the prepared leaf extract was evaluated against early fourth instar larvae of Ae. aegypti using WHO protocol. The mortality counts were made after 24 h and LC50 and LC90 values were calculated. The efficacy of extract as mosquito irritant was assessed by contact irritancy assays. Extract-impregnated paper was placed on a glass plate over which a perspex funnel with a hole on the top was kept inverted. Single female adult, 3-day old unfed/blood-fed, was released inside the funnel. After 3 min of acclimatization time, the time taken for the first take-off and total number of flights undertaken during 15 min were scored. RESULTS: The citrus leaf extracts from hexane possessed moderate larvicidal efficiency against dengue vector. The bioassays resulted in an LC50 and LC90 value of 446.84 and 1 370.96 ppm, respectively after 24 h of exposure. However, the extracts were proved to be remarkable irritant against adults Ae. aegypti, more pronounced effects being observed on blood-fed females than unfed females. The extract-impregnated paper was thus proved to be 7-11 times more irritable as compared with the control paper. CONCLUSIONS: The hexane extracts from C. sinensis leaves are proved to be reasonably larvicidal but remarkably irritant against dengue vector. Further studies are needed to identify the possible role of extract as adulticide, oviposition deterrent and ovicidal agent. The isolation of active ingredient from the extract could help in formulating strategies for mosquito control.

Bioefficacy of Mentha piperita essential oil against dengue fever mosquito Aedes aegypti L.

OBJECTIVE: To assess the larvicidal and repellent potential of the essential oil extracted from the leaves of peppermint plant, Mentha piperita (M. piperita) against the larval and adult stages of Aedes aegypti (Ae. Aegypti). METHODS: The larvicidal potential of peppermint oil was evaluated against early fourth instar larvae of Ae. aegypti using WHO protocol. The mortality counts were made after 24 and 48 h, and LC50 and LC90 values were calculated. The efficacy of peppermint oil as mosquito repellent was assessed using the human-bait technique. The measured area of one arm of a human volunteer was applied with the oil and the other arm was applied with ethanol. The mosquito bites on both the arms were recorded for 3 min after every 15 min. The experiment continued for 3 h and the percent protection was calculated. RESULTS: The essential oil extracted from M. piperita possessed excellent larvicidal efficiency against dengue vector. The bioassays showed an LC50 and LC90 value of 111.9 and 295.18 ppm, respectively after 24 h of exposure. The toxicity of the oil increased 11.8% when the larvae were exposed to the oil for 48 h. The remarkable repellent properties of M. piperita essential oil were established against adults Ae. aegypti. The application of oil resulted in 100% protection till 150 min. After next 30 min, only 1-2 bites were recorded as compared with 8-9 bites on the control arm. CONCLUSIONS: The peppermint essential oil is proved to be efficient larvicide and repellent against dengue vector. Further studies are needed to identify the possible role of oil as adulticide, oviposition deterrent and ovicidal agent. The isolation of active ingredient from the oil could help in formulating strategies for mosquito control.