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The Essential Oil of Lippia Alba Mill (Lamiales:Verbenaceae) As Mosquitocidal and Repellent Agent Against Culex Quinquefasciatus

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The Essential Oil of Lippia Alba Mill (Lamiales:Verbenaceae) As Mosquitocidal and Repellent Agent Against Culex Quinquefasciatus Mahanta et al. The Journal of Basic and Applied Zoology (2019) 80:64 The Journal of Basic https://doi.org/10.1186/s41936-019-0132-0 and Applied Zoology RESEARCH Open Access The essential oil of Lippia alba Mill (Lamiales:Verbenaceae) as mosquitocidal and repellent agent against Culex quinquefasciatus Say (Diptera: Culicidae) and Aedes aegypti Linn (Diptera: Culicidae) Sudarshana Mahanta, Riju Sarma and Bulbuli Khanikor* Abstract Background: Being a low-risk insecticide, plant essential oils emerge as competent mosquitocidal and repellent candidates. However, essential oil may act differently in different mosquito species and different developmental stages of same mosquito species. In the current investigation, we evaluated the ovicidal, larvicidal, adulticidal and repellent activities of essential oil extracted from the leaves of Lippia alba against two medically important mosquito species, Aedes aegypti and Culex quinquefasciatus. The efficacy of the oil was assessed under laboratory conditions against different developmental stages of the selected species. Results: Fromthefindings,itcanbeinferredthatL. alba oil is more effective as a repellent against both the targeted mosquito species. Results also demonstrated that ovicidal, larvicidal and repellent properties of the essential oil were higher against Culex quinquefasciatus than Aedes aegypti. GC-MS analysis of the oil showed the presence of aromadendrene oxide, caryophyllene oxide, etc. as major constituent compounds. Conclusion: The outcomes of this study clearly indicated that the essential oil of L. alba has the potentiality to act more effectively as a repellent, followed by a larvicidal agent against mosquito and at the same time the results revealed differential vulnerability of different mosquito species and their life stages against a particular plant essential oil. Keywords: Lippia alba, Essential oil, Aedes aegypti, Culex quinquefasciatus Background value have been extensively studied in different countries Repeated use of synthetic insecticides for controlling mos- across the world against diverse pests including mosqui- quitoes has created severe trouble like resurgences of toes. Researchers are also observing the combined efficacy mosquitoes and undesirable effects on non-target organ- of the plant-based product and bacteria-based insecticide isms. It has also fostered environmental and human health with chemical insecticides to increase the effectiveness of concerns and led to the development of resistance in mos- a vector control strategy (Nassar and El-Sebaii, 1999;Nas- quito populations. Therefore, it is needful to generate sar, Ghramh, Al-Wazi, Mahyoub, Ahmed, 2017). How- some novel insecticides which have low mammalian tox- ever, detail studies on some of the commonly found icity, eco-friendliness and at the same time effective in kill- aromatic plants are still lacking which can be effective ing mosquitoes or to prevent biting of mosquitoes. A against mosquito vectors. number of natural products in the form of plant extracts, Lippia is one of the important genera of the family plant essential oils and compounds having insecticidal Verbenaceae which comprises around 220 species (Ngas- sapa, Runtoro, Harvala, and Chinou, 2003). Among these, * Correspondence: [email protected] Lippia alba (Verbenaceae) is an aromatic, perennial shrub, Department of Zoology, Gauhati University, Guwahati, Assam 781014, India © The Author(s). 2019 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. Mahanta et al. The Journal of Basic and Applied Zoology (2019) 80:64 Page 2 of 7 found in different countries including India. In Collection of plant material Assam, the plant is widely grown and locally known Leaves of Lippia alba were collected from Jalukbari, as “pikhachi bon”. Traditionally the plant is used as Guwahati, Assam, India. Plant materials were collected spice among different tribes in the state. Some of the in the period of March to June 2018. Collected speci- researchers have explored several major compounds mens were identified in the Department of Botany, Gau- from L. alba essential oils which were found to pos- hati University, Assam, India. sess anti-viral, anti-bacterial, anti-fungal activities (Rahmatillah et al. 2011). The efficacy of essential oil Essential oil extraction from leaves of L. alba has not been studied widely Fresh plant materials were taken for the extraction of es- against mosquitoes, although the repellent activity was sential oil and no solvent was used in it. The essential established against A. aegypti (Castillo, Stashenko, and oil was extracted from fresh the plant materials with the Duque, 2017). Besides this, the repellent property of help of Clevenger apparatus by the hydro-distillation the plant was also reported against stored grain pests method described by Kumar, Kumar, Prasal, Dubey, and (Peixoto et al. 2015). Keeping it on the mind, a de- Samant (2008) with little modification. Five hundred tailed study on the efficacy of L. alba (leaves) oil was grams of plant material was taken for the extraction. done against all the developmental stages of C. quin- The apparatus was run for 6 h after boiling of water. quefasciatus and A. aegypti which is not found to be The extracted essential oil was collected in glass vials, reported yet. few crystals of anhydrous sodium sulphate were added In addition, vector control measures will be successful to absorb excess water present in collected plant oil and if we are able to identify the selective toxicity of a plant the vial was stored in 4° centigrade. essential oil towards different mosquito species as well as their different developmental stages. This is because a Activity of essential oils variety of mosquito species with their different develop- Ovicidal activity mental stages may be present together in a particular lo- The ovicidal bioassay was performed according to the cation where mosquitocides are applied and or method described by Tennyson, Ravindran, and Arivoli dominance of a particular mosquito species may be (2011) and Pushpanathan, Jebanesan, and Govindarajan higher in a particular geographical locality. Insufficiency (2006) with little modifications. Initially, two concentra- in the knowledge of the toxicity of controlling agent and tions (1000 ppm and 500 ppm) were used to observe its its species specificity, the control measures would not effect and based on the findings; six different concentra- give expected results. The same essential oil may not tions (1000 ppm, 500 ppm, 250 ppm, 100 ppm, 50 ppm, have a similar activity towards different mosquito spe- 10 ppm) were applied to determine the sublethal con- cies. Even it does not affect equally against different de- centration (LC50). Fifty eggs of each of the mosquito velopmental stages of a single species. To broaden the species were exposed to each concentration of essential information of the mosquitocidal activity of L. alba oil, oil. Dimethyl sulfoxide (DMSO) was used as an emulsi- an attempt was made in the current investigation to ac- fying agent for the preparation of each concentration. cess its efficacy against different developmental stages of Each experiment was replicated thrice along with appro- Aedes aegypti and Culex quinquefasciatus. priate control (DMSO-treated water as positive control and water as the negative control). Materials and methods Rearing of A. aegypti and C. quinquefasciatus Percent ovicidal activity 1 The eggs of A. aegypti and C. quinquefasciatus were col- ¼ %of eggs hatched in control − %eggs hatched in treated=% A Â 100 lected from Regional Medical Research Laboratory of eggs hatched in control (RMRC), Dibrugarh, Assam, India. The collected eggs were reared in the insect culture room, Department of Zoology, Gauhati University, Assam, India, by following Larvicidal bioassay the rearing practices described by Arivoli and Tennyson The efficacy of essential oil of L. alba was studied by (2011). The colonies were maintained at the temperature performing bioassay against third instar larval stages of between 25 and 29 °C temperature and 80–90% relative A. aegypti and C. quinquefasciatus. The larvicidal activity humidity. Larvae were fed on larval food (powdered dog of the essential oil was assayed following the method de- biscuit and yeast in the ratio 3:1) and adult mosquitoes scribed by Tong and Bloomquist (2013). For larvicidal were fed on 10% glucose solution. Pupae were trans- bioassay, batches of 20 numbers of healthy larvae were ferred to a disposable cup and it was kept inside the transferred to the disposable cups (depth 10 cm) con- cage. On the fifth day after hatching, adult female mos- taining 100 ml of water. At first, two concentrations quitoes were blood-fed on albino rats. (1000 ppm and 500 ppm) were used to observe its Mahanta et al. The Journal of Basic and Applied Zoology (2019) 80:64 Page 3 of 7 efficacy after which six different concentrations (1000 served as control. Each bioassay was performed be- ppm, 500 ppm, 250 ppm, 100 ppm, 50 ppm, 10 ppm) tween 17:00 and 20:00 h for C. quinquefasciatus and were applied. Test solutions were prepared by mixing an for A. aegypti it was tested between 9:00 and 12:00 h. equal
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