LARVICIDAL ACTIVITY OF EPALTES PYGMAEA (DC) WHOLE PLANT EXTRACTS AGAINST CULEX QUINQUEFASCIATUS SAY, AEDES AEGYPTI L. AND ANOPHELES STEPHENSI LISTON LARVAEHTML Full Text
LARVICIDAL ACTIVITY OF EPALTES PYGMAEA (DC) WHOLE PLANT EXTRACTS AGAINST CULEX QUINQUEFASCIATUS SAY, AEDES AEGYPTI L. AND ANOPHELES STEPHENSI LISTON LARVAE
S. Murugammal 1, M. Rajiv Gandhi 2, P. Pandikumar 2, R. Shakila 3 and R. Ilavarasan * 1
Captain Srinivasa Murti Reseach Institute for Ayurveda Drug Development 1, Anna Hospital Campus, Arumbakkam, Chennai - 600106, Tamil Nadu, India.
Division of Vector Control 2, Entomology Research Institute, Loyolo College, Chennai - 600034, Tamil Tamil Nadu, India.
Siddha Central Research Institute (Central Council for Research in Siddha) 3, Anna Hospital Campus, Arumbakkam, Chennai - 600106, Tamil Nadu, India.
ABSTRACT: In the present study, the larvicidal activity of successive n-hexane, chloroform, ethyl acetate and ethanol extracts of Epaltes pygmaea DC (Family: Asteraceae) whole plant was carried out against three mosquito species namely Culex quinquefasciatus, Aedes aegypti L. and Anopheles stephensi. The crude whole plant was successively extracted with n-hexane, chloroform, ethyl acetate, and ethanol. The larvicidal activity was studied at 62.5 ppm, 125 ppm, 250 ppm, and 500 ppm concentrations against the late third instar larvae of C. quinquefasciatus, A. aegypti and A. stephensi. Results showed that the ethyl acetate extract of E. pygmaea was the most effective against tested mosquito larvae. The median lethal concentration (LC50) values of ethyl acetate extract were calculated as 35.79 ppm, 62.37 ppm and 29.94 ppm for C. quinquefasciatus, A. aegypti and A. stephensi, respectively. The results suggested that ethyl acetate extract of E. pygmaea could be used to control C. quinquefasciatus, A. aegypti and A. stephensi.
Epaltes pygmaea, Larvicidal, Bioassay, Jaundice
INTRODUCTION: Epaltes pygmaea DC is a small annual herb, 8 to 20 cm high, minutely winged branched stem with aromatic roots, leaves are alternate, linear, lanceolate to oblong, flower color is pink, solitary, terminal, heterogamous. It is found in Sri Lanka, India, Java and China and a limited extent in South India, especially towards the coast, gregarious in low lying ground by river banks and paddy field after harvesting in clayey soil 1-3.
Epaltes is used in traditional Ayurvedic medicine in Srilanka to alleviate jaundice. A literature survey reveals that the plant of the genus has the therapeutic action of diaphoretics, diuretics, stimulant, and expectorant are used in urethral discharges and acute dyspepsia 4. Alcohol and aqueous extract of E. pygmaea possesses hepatoprotective activity against paracetamol-induced liver damage in rats and also have potent diuretic activity 5.
Recent studies show that the extract of E. pygmaea has good inhibitory activity against the organism Bacillus cereus, Klebsiella pneumonia and Staphylococcus aureus at microgram level 6. Microscopic, thin layer chromatographic studies 7 and quality control parameters 8 of E. pygmaea have been reported.
The present study was conducted to evaluate the larvicidal activity of the plant Epaltes pygmaea DC. whole plant extracts against the larvae of C. quinquefasciatus, A. aegypti and A. stephensi.
MATERIALS AND METHODS:
Plant Collection: Fresh whole plant of Epaltes pygmaea DC. was collected from Tirunelveli District in September 2016 was identified and authenticated by Prof. P. Jeyaraman, Director, Institute of Herb Botany, Plant Anatomy Research Centre, Tambaram, Chennai, India, where a voucher specimen (PARC/2014/2071) was deposited. The plant was dried in the shade and powdered in a pulverizer.
Extraction: The plant material was extracted with n-hexane, chloroform, ethyl acetate, and ethanol solvents in a successive manner using a Soxhlet apparatus. All the obtained extracts were filtered using sterile Whatman filter paper no. 2, dried using Rotavapor R-300 and stored separately for further use. The percentage yield of hexane, chloroform, ethyl acetate, and ethanol were 5.3, 3.7, 4.5 and 6.5 respectively.
Mosquitoes: Laboratory reared C. quinquefasciatus, A. aegypti and A. stephensi mosquito larvae were used in the bioassay experiments. The rearing conditions were: 27 ± 1 ºC, 75-85% RH and 14:10 h photoperiod.
Mosquito Larvicidal Bioassay: Larvicidal activity of the extracts was evaluated using the method recommended by the World Health Organization 9 with slight modifications. A range of concentrations viz., 62.5, 125, 250 and 500 ppm of each solvent extract were prepared with an emulsifier and taken in 100 ml plastic cups separately. Third instar larvae (20 larvae) of each A. aegypti, C. quinquefasciatus and A. stephensi were introduced into each cup that contained test solutions. Emulsifier control and water control were also maintained separately. Five replicates were maintained for each treatment and control. Larval mortality was recorded after 24 h of treatment. Larvae were considered dead when they did not move or rise to the surface of the solution.
Statistical Analysis: The lethal concentration values (LC50 and LC90) were calculated by EPA probit analysis software (1.5 versions) 10.
RESULTS AND DISCUSSION: Larvicidal effect of E. pygmaea whole plant extracts on the third instar larvae was recorded and the results are presented in Table 1. The ethyl acetate extract of E. pygmaea was very effective against all three mosquito larvae. The ethyl acetate extract of E. pygmaea was most effective against the larvae of A. stephensi with LC50 and LC90 values of 29.94, 141.0 ppm. The LC50 and LC90 values of ethyl acetate extract of E. pygmaea were 35.79, 160.47 ppm and 62.37, 313.81 ppm against the larvae of C. quinquefasciatus and A. aegypti, respectively.
The LC50 and LC90 values of E. pygmaea hexane extract were 247.72 and 1599.42 ppm against C. quinquefasciatus and 495.28 and 4288.45 ppm against A. aegypti larvae and 168.50 and 1640.68 ppm against A. stephensi larvae, respectively Table 1. The LC50 and LC90 values of E. pygmaea chloroform extract were 40.99 and 197.81 ppm against C. quinquefasciatus, 69.12 and 433.221 ppm against A. aegypti and 32.623 and 174.110 ppm against Anopheles stephensi larvae, respectively Table 1. The LC50 and LC90 values of E. pygmaea ethanol extract were 350.25 and 2912.24 ppm against C. quinquefasciatus, 485.12 and 4937.64 ppm against A. aegypti larvae and 244.55 and 1994.81 ppm against A. stephensi larvae, respectively Table 1.
The active ethyl acetate extract of E. pygamea extract contained terpenoids, flavonoids, phenol, tannin, steroid group of phytochemicals; the mortality may be due to these phytochemicals. Culex quinquefasciatus control was mainly conducted through the use of neurotoxic insecticides belonging to the organochlorines (OC), the organophosphates (OP) and the pyrethroids 11.
Aedes aegypti is the main vector of Dengue and Dengue hemorrhagic fevers 12, 13. Anopheles stephensi larvae is a major malaria vector 14. Hence researchers continuously study to control these larvae with herbal extracts and found Tridax procumbens L. 15 and Leucas aspera L. 16 with positive results. Results from the present study also suggested E. pygmaea was found effective in controlling of C. quinquefasciatus, A. aegypti, and A. stephensi and successive ethyl acetate extract as most active than other extracts.
TABLE 1: LETHAL CONCENTRATION (IN PPM) OF VARIOUS EXTRACTS OF E. PYGMAEA AGAINST THREE MOSQUITO LARVAE
|95% confidence limit||95% confidence limit|
|Mosquito species||Treatment||LC50 (ppm)||LL||UL||LC90 (ppm)||LL||UL||Slope ± SE||Intercept ± SE||χ2|
|Hexane||247.72||189.47||350.91||1599.42||869.086||5538.65||1.58 ± 0.28||1.21 ± 0.66||0.1*|
|Chloroform||40.99||17.43||61.04||197.81||147.60||327.77||1.87 ± 0.40||1.97 ± 0.84||1.9*|
|Ethyl acetate||35.79||13.27||54.86||160.47||120.57||254.70||1.96 ± 0.44||1.94 ± 0.92||1.9*|
|Ethanol||350.25||253.32||611.18||2912.24||1276.08||19685.18||1.39 ± 0.29||1.45 ± 0.67||0.1*|
|Hexane||495.28||336.59||1093.27||4288.45||1647.23||45487.46||1.36± 0.30||1.31 ± 0.70||0.2*|
|Chloroform||69.12||38.68||95.52||433.221||294.112||926.839||1.60± 0.31||2.04 ± 0.68||3.0*|
|Ethyl acetate||62.37||36.05||84.97||313.81||227.77||560.07||1.82± 0.34||1.72 ± 0.73||5.6*|
|Ethanol||485.12||323.76||1152.39||4937.64||1743.16||74921.77||1.27 ± 0.29||1.58 ± 0.68||0.1*|
|Hexane||168.50||117.37||237.61||1640.68||802.33||8806.38||1.29 ± 0.27||2.11 ± 0.63||0.1*|
|Chloroform||32.62||9.942||52.950||174.110||127.582||293.164||1.76 ± 0.41||2.33± 0.87||1.8*|
|Ethyl acetate||29.94||8.18||49.19||141.00||103.65||223.53||1.90± 0.47||2.18 ± 0.97||1.5*|
|Ethanol||244.55||181.52||364.69||1994.81||972.12||9872.57||1.40 ± 0.28||1.64± 0.65||0.6*|
LC50: Lethal concentration to kill 50% of the exposed larvae; LC90: Lethal concentration to kill 90% of the exposed larvae; P=≤0.05-chi-square values were significant, LL: Lower Limit, UL: Upper Limit.
CONCLUSION: In conclusion, the ethyl acetate extract of E. pygmaea was the most potent treatment against the three tested mosquito vectors. Based on these results, the ethyl acetate extract of E. pgymaea could be used in vector mosquito control and maybe further investigated to isolate the active constituent responsible for the larvicide.
ACKNOWLEDGEMENT: The authors are thankful to Entomology Research Institute, Loyola College, Chennai, providing necessary facilities.
CONFLICTS OF INTEREST: There is no conflict of interest.
- Mathew KM: An exclusive flora of Central Tamil Nadu, The Ranipat Herbarium, St. John’s College, Tiruchirapalli. India, Published by CRC press 1995: 259.
- Mayuranathan PV: The flowering plants of Madras city and its immediate neibourhoodk Revised by Livingstone C and Henry AN. New series-Natural History Section 1994; 10: 153.
- Pullaiah T, Ramamurthy KS and Karuppusamy S: Flora of Eastern Ghats, Hill Ranges of South East India 2001; 3: 264.
- Mukerjee PK: Quality control of Herbal Drugs – An approach to evaluation of Botanicals, Business Horizons, Pharmaceutical publishers, New Delhi 2008: 2-11.
- Amala K, Saraswathy A and Amerjothy S: Hepatoprotective and diuretic activities of alcoholic and aqueous extracts of Epalte spygmaea DC. (Asteraceae). Int J Pharmacy and Pharmaceutical Sci 2013; 5(2): 502-06.
- Murugammal S, Sri PM, Shakila R and Ilavarasan R: Exploration of antibacterial activity of various extracts of Epaltes pygmaea Against few bacterial organisms. Int J Res Ayurveda Pharm. 2018; 9(2): 52-56.
- Amala K, Ilavarasan R and Amerjothy S: Standardization of plant Epaltes pygmaea (Asteracea). Journal of Pharmacy Research 2016; 10(10): 680-82.
- Amala K, Ilavarasan R and Amerjothy S: Development of Quality control standards for two Epaltes species. International J of Green Pharmacy 2017; 11(1): S94-S99.
- Anonymous: Guidelines for laboratory and field testing of mosquito larvicides. WHO, Geneva, WHO/CDS/ WHOPES/GCDPP/13, 2005.
- Finney DJ: Probit analysis, Cambridge University Press, London 1971: 256.
- Bhattacharya S and Basu P: The Southern House Mosquito, Culex quinquefasciatus: profile of a smart vector J of Entomol and Zool Studies 2016; 4(2): 73-81.
- Getachew D, Tekie H, Michael TG, Balkew M and Mesfin A: Breeding Sites of Aedes aegypti: Potential Dengue Vectors in Dire Dawa, East Ethiopia. Interdisciplinary Perspectives on Infectious Diseases 2015; 1-8.
- Lenhart AE, Walle M, Cedillo H and Kroeger A: Building better ovitraps for detecting Aedes aegypti oviposition. Acta Tropica 2005; 96: 5659.
- Surendran SN, Sivabalakrishnan K, Gajapathy K, Arthiyan S, Jayadas TTP, Karvannan K, Raveendran S, Karunaratne SH PP and Ramasamy R: Genotype and biotype of invasive Anopheles stephensi in Mannar Island of Sri Lanka. Parasites & Vectors 2018; 11(3): 1-7.
- Elumalai D, Kaleena PK, Fathima M and Nareshkumar C: Phytochemical screening and larvicidal activity of procumbens (L) against A. stephensi (Liston), A. aegypti (L) and C. quinquefasciatus (say). IJBR 2013; 2: 1-14.
- Elumalai D, Hemalatha P and Kaleena PK: Larvicidal activity and GC–MS analysis of Leucas aspera against Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus. J Saudi Soc Agric Sci 2017; 16: 306-13.
How to cite this article:
Murugammal S, Gandhi MR, Pandikumar P, Shakila R and Ilavarasan R: Larvicidal activity of Epaltes pygmaea (DC) whole plant extracts against Culex quinquefasciatus say, Aedes aegypti L. and Anopheles stephensi Liston larvae. Int J Pharm Sci & Res 2020; 11(3): 1296-98. doi: 10.13040/IJPSR.0975-8232.11(3).1296-98.
All © 2013 are reserved by the International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.
S. Murugammal, M. R. Gandhi, P. Pandikumar, R. Shakila and R. Ilavarasan *
Department of Life Sciences, CHRIST (Deemed to be University), Bengaluru, Karnataka, India.
09 May 2019
25 October 2019
13 November 2019
01 March 2020