Sarita Kumar

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.

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.

Microplate assay of elevated esterase activity in individual pyrethroid-resistant mosquitoes

Involvement of esterase-mediated hydrolysis as a mechanism of pyrethroidresistance in three species of mosquitoes,viz., Aedes aegypti, Culex quinquefasciatus andAnopheles Stephensi was investigated by microplate assay of Β-esterases in individual larva and adult female and male mosquitoes. Assuming an absorbance value of 0.4 and above at 555 nm as the threshold level of elevated esterase activity which confers resistance, frequency distributions of such individual test mosquitoes were constructed in resistant and susceptible populations. The results indicate the involvement of ester hydrolysis of Pyrethroids as a predominant mechanism of pyrethroid-resistance in the larvae ofCulex quinquefasciatus but not inAedes aegypti. However, a marginal role of esterases is indicated in the larvae ofAnopheles stephensi.

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 LC50 value of 30.71 ppm while the Phyllanthus emblica fruit extract was found to be least effective with an LC50 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 LC50 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.

Evaluation of the larvicidal efficiency of stem, roots and leaves of the weed, Parthenium hysterophorus (Family: Asteraceae) against Aedes aegypti L.

Abstract Objective To assess the larvicidal potential of various extracts prepared from the stem, roots and leaves of Parthenium hysterophorus ( P. hysterophorus ) against 3rd and 4th instars of Aedes aegypti (Ae. aegypti) . Methods The extracts from each part were prepared with four solvents; petroleum ether, hexane, acetone and diethyl ether. Each part was dried, powdered and soaked in different solvents, separately, for five days. The crude extracts thus formed were concentrated using rotary evaporator and stored as stock solution of 1 000 mg/L. Results All the extracts prepared from the leaves were found ineffective against both the instars causing only 10%-40% mortality. Against 3rd instars, the hexane and petroleum ether extracts prepared from the stem of P. hysterophorus were found effective exhibiting LC 50 values of 379.76 and 438.57 mg/L, respectively. Likewise the hexane and petroleum ether extracts from the Parthenium roots resulted in LC 50 values of 432.38 and 562.50 mg/L, respectively, against 4th instars of Ae. aegypti revealing their larvicidal potential. It was further found that the hexane extracts, whether from roots or stem, were 13-28% more effective than the petroleum ether extracts. The qualitative phytochemical study of the effective extracts from the stems and roots showed the presence of alkaloids, saponins, terpenoids and flavonoids in different combinations. Conclusions Our investigations demonstrated the potential of P. hysterophorus roots and stems against Ae. aegypti larvae and their benefits as new types of mosquito larvicides. Variety of types and levels of active constituents in each kind of extract may be responsible for the variability in their potential against Ae. aegypti . Further research is needed to identify these components.

Multiple insecticide resistance/susceptibility status of Culex quinquefasciatus, principal vector of bancroftian filariasis from filaria endemic areas of northern India.

OBJECTIVE To understand the insecticide resistance status of Culex quinquefasciatus Say (Diptera: Culicidae) (Cx. Quinquefasciatus) to deltamethrin, cyfluthrin, permethrin, lambdacyhalothrin, DDT and malathion in filarial endemic areas of Uttar Pradesh, India. METHODS Insecticide susceptibility assays were performed on wild-caught adult female Cx. quinquefasciatus mosquitoes to deltamethrin (0.05%), cyfluthrin (0.15%), permethrin (0.75%), lambdacyhalothrin (0.05%), malathion (5.0%) and DDT (4.0%), the discriminating doses recommended by the World Health Organisation (WHO). RESULTS The data showed that Cx. quinquefasciatus is highly resistant to DDT and malathion; the mortality was 28.33% and 27.5%, respectively and incipient resistance to synthetic pyrethroids (deltamethrin, cyfluthrin, permethrin, and lambdacyhalothrin), where mortality ranged from 95.83% in permethrin to 98.33% in cyfluthrin and lambdacyhalothrin. Knockdown times (KDT(50)) in response to synthetic pyrethroids varied significantly between different insecticides (P<0.01) from 31.480 min for permethrin to 21.650 for cyfluthrin. CONCLUSIONS The results presents here provide the status report of the insecticide resistance/susceptibility of Cx. quinquefasciatus in major filaria endemic areas of northern India.

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.

Deltamethrin: Promising mosquito control agent against adult stage of Aedes aegypti L.

OBJECTIVE To evaluate the effects of deltamethrin against field-collected adults of Aedes aegypti L. (Ae. aegypti). METHODS The adults were selected with 0.025% deltamethrin for 40 successive generations. The selected adults were tested with 4% DDT and the emerging larvae were tested with various insecticides to study the cross-resistance spectrum. The knockdown and irritability studies were carried out in adult mosquitoes to investigate their behavioural response to deltamethrin. RESULTS Forty generations of selection with deltamethrin resulted in only 3.8-fold resistance in the adults of Ae. aegypti. The adults of parent (PS) and selected strains (DAS) exhibited only 0.8-fold cross resistance to 4% DDT. The larvae emerging from the PS and DAS strains did not develop appreciable levels of resistance to various insecticides tested. The knockdown studies revealed KT50 of 14.4 min in PS adults with no signs of recovery even after 24 h. The DAS strains could develop only 1.2 to 1.3-fold knockdown resistance (KDR). The knockdown response of DDT was though 5-6 times slower than deltamethrin but the continued response in deltamethrin-selected adults caused only 1.2-fold KDR. The PS and DAS strains exhibited significant irritability response towards deltamethrin and DDT. The DAS strains showed 5-6 fold increased irritability to deltamethrin as compared to the PS strain. CONCLUSIONS The above results suggest the prolonged effective use of deltamethrin against Ae. aegypti as an adulticide.

Evaluation of the Larvicidal Efficacy of Five Indigenous Weeds against an Indian Strain of Dengue Vector, Aedes aegypti L. (Diptera: Culicidae)

Background and Objectives. Aedes aegypti, dengue fever mosquito, is primarily associated with the transmission of dengue and chikungunya in tropical and subtropical regions of the world. The present investigations were carried out to assess the larvicidal efficiency of five indigenous weeds against Ae. aegypti. Methods. The 1,000 ppm hexane and ethanol extracts prepared from the leaves and stem of five plants (Achyranthes aspera, Cassia occidentalis, Catharanthus roseus, Lantana camara, and Xanthium strumarium) were screened for their larvicidal activity against early fourth instars of dengue vector. The extracts which could cause 80–100% mortality were further investigated for their efficacy. Results. The preliminary screening established the efficacy of hexane extracts as compared to the ethanol extracts. Further investigations revealed the highest larvicidal potential of A. aspera extracts exhibiting LC50 value of 82.555 ppm and 68.133 ppm, respectively. Further, their leaf extracts showed 5–85.9% higher larvicidal activity and stem extracts exhibited 0.23- to 0.85-fold more efficiency than the other four extracts. Conclusion. The present investigations suggest the possible use of A. aspera as an ideal ecofriendly, larvicidal agent for the control of dengue vector, Ae. aegypti. Future studies are, however, required to explore and identify the bioactive component involved and its mode of action.

Impact of the Stem Extract of Thevetia neriifolia on the Feeding Potential and Histological Architecture of the Midgut Epithelial Tissue of Early Fourth Instars of Helicoverpa armigera Hübner

Helicoverpa armigera HÜbner is one of the most important agricultural crop pests in the world causing heavy crop yield losses. The continued and indiscriminate use of synthetic insecticides in agriculture for their control has received wide public apprehension because of multifarious problems, including insecticide resistance, resurgence of pest species, environmental pollution, and toxic hazards to humans and nontarget organisms. These problems have necessitated the need to explore and develop alternative strategies using eco-friendly and biodegradable plant products. In view of this, the efficacy of Thevetia neriifolia methanol stem extract was evaluated against the early fourth instars of H. armigera as an antifeedant and stomach poison agent. Feeding of larvae with the diet containing 0.005%–5.0% extract resulted in 2.06%–37.35% antifeedant index; the diet with 5.0% extract caused 54.3% reduced consumption. The negative impact of extract on larval feeding resulted in 37.5%–77.7% starvation, causing adverse effects on the larval weight. Choice between control and experimental diet resulted in feeding preference of larvae for the control diet, leading to 7.3%–42.9% reduced consumption of extract-containing diet. The only exception was the diet with 0.005% extract, which could not cause any deterrence. The midgut histological architecture of H. armigera larvae fed with 0.005%–0.05% extract-containing diet with negligible antifeedant potential showed significant damage, shrinkage, and distortion and vacuolization of gut tissues and peritrophic membrane, causing the disintegration of epithelial, goblet, and regenerative cells; the damage increased with the increase in concentration. These changes in the gut caused negative impact on the digestion and absorption of food and thus nutritional deficiency in the larvae, which could probably affect their growth and development. This study reveal the appreciable stomach poison potential of T. neriifolia stem methanol extract against H. armigera larvae, which can be explored as an eco-friendly pest control strategy.