Screening for Larvicidal Activity of Aqueous and Ethanolic Extracts of Fourteen Selected Plants and Formulation of a Larvicide against Aedes aegypti (Linn.) and Aedes albopictus (Skuse) Larvae
This study aims to: a) obtain ethanolic (95% EtOH) and aqueous extracts of Selaginella elmeri, Christella dentata, Elatostema sinnatum, Curculigo capitulata, Euphorbia hirta, Murraya koenigii, Alpinia speciosa, Cymbopogon citratus, Eucalyptus globulus, Jatropha curcas, Psidium guajava, Gliricidia sepium, Ixora coccinea and Capsicum frutescens and screen them for larvicidal activities against Aedes aegypti (Linn.) and Aedes albopictus (Skuse) larvae; b) to fractionate the most active extract and determine the most active fraction; c) to determine the larvicidal properties of the most active extract and fraction against by computing their percentage mortality, LC50, and LC90 after 24 and 48 hours of exposure; and d) to determine the nature of the components of the active extracts and fractions using phytochemical screening. Ethanolic (95% EtOH) and aqueous extracts of the selected plants will be screened for potential larvicidal activity against Ae. aegypti and Ae. albopictus using standard procedures and 1% malathion and a Piper nigrum based ovicide-larvicide by the Department of Science and Technology as positive controls. The results were analyzed using One-Way ANOVA with Tukey’s and Dunnett’s test. The most active extract will be subjected to partial fractionation using normal-phase column chromatography, and the fractions subsequently screened to determine the most active fraction. The most active extract and fraction were subjected to dose-response assay and probit analysis to determine the LC50 and LC90 after 24 and 48 hours of exposure. The active extracts and fractions will be screened for phytochemical content. The ethanolic extracts of C. citratus, E. hirta, I. coccinea, G. sepium, M. koenigii, E globulus, J. curcas and C. frutescens exhibited significant larvicidal activity, with C. frutescens being the most active. After fractionation, the ethyl acetate fraction was found to be the most active. Phytochemical screening of the extracts revealed the presence of alkaloids, tannins, indoles and steroids. A formulation using talcum powder–300 mg fraction per 1 g talcum powder–was made and again tested for larvicidal activity. At 2 g/L, the formulation proved effective in killing all of the test larvae after 24 hours.
[1] Tayag, E. A., Roque, Jr., V. G., and Elfa, D. C. Disease surveillance report. Retrieved from: www.doh.gov.ph/sites/default/files/leptoup21.pdf, 201.
[2] World Health Organization. Dengue: Guidelines for diagnosis, treatment, prevention and control. Retrieved from: www.who.int/tdr/publications/documents/dengue-diagnosis.pdf, 2009.
[3] Negrelle, R. R. B., and Gomes, E. C. Cymbopogon citratus (DC.) Stapf: Chemical composition and biological activities. Rev. Bras. Pt. Med., 9(1), 80-92. Retrieved from: http://www.sbpmed.org.br/download/issn_07_1/revisao_v9_n1.pd,2007.
[4] Karunamoorthi, K., and Ilango, K. Larvicidal activity of Cymbopogon citratus (DC Stapf. and Croton macrostachyus Del. against Anopheles arabiensis Patton, a potent malarial vector. European Review for Medical and Pharmacological Sciences, 14, 57-62. Retrieved from: http://www.ncbi.nlm.nih.gov/pubmed/20184090, 2007.
[5] Cavalcanti, E., de Morais, S., Lima, M. A., & Santana, E. Larvicidal Activity of Essential Oils from Brazilian Plants against Aedes aegypti L. Mem. Inst. Oswaldo Cruz, 99(5), 541-544. Retrieved from: http://www.ncbi.nlm.nih.gov/pubmed/15543421, 2004.
[6] Quisumbing, E. Medicinal Plants of the Philippines. Quezon City, Philippines: JMC Press, Inc., 1978.
[7] Madhumathy, A., Aivazi, A. A., and Vijayan, V. Larvicidal efficacy of Capsicum annum against Anopheles stephensi and Culex quinquefasciatus. Journal of Vecotr Borne Diseases, 44, 223-226. Retrieved from: http://www.ncbi.nlm.nih.gov/pubmed/17896626, 2007.
[8] Vinayaka, K. S., Prakish Kekuda, T. R. and Nandini, K.C. Potent insecticidal activity of fruits and leaves of Capsicum frutescens (L.) var. longa (Solanaceae). Der Pharmacia Lettre, 2(4), 172-176. Retrieved from: http://scholarsresearchlibrary.com/DPL-vol2-iss4/DPL-2010-2-4-172-176.pdf, 2010.
[9] Batish, D. R., Singh, H. P., Kohli, R. K., & Kaur, S. Eucalyptus essential oil as a natu ral pesticide. Forest Ecology and Management, 256, 2166-2174. doi: 10.1016/j.foreco.2008.08.008, 2008.
[10] Kumar, S., Malhotra, R., and Kumar, D. Euphorbia hirta: Its chemistry, traditional and medicinal uses, and pharmacological activities. Pharmacgon. Rev., 4(7), 58-61. doi: 10.4103/0973-7847.65327, 2010.
[11] Patil, S., Patil, C., Salunkhe, R., & Salunke, B. Larvicidal activities of six plants extracts against two mosquito species, Aedes aegypti and Anopheles stephensi. Tropical Biomedicine, 27(3), 360-365. Retrieved from: http://www.ncbi.nlm.nih.gov/pubmed/21399575, 2010.
[12] Uche, F., and Aprioku, J. The Phytochemical Constituents, Analgesic and Anti-inflammatory effects of methanol extract of Jatropha curcas leaves in Mice and Wister albino rats. J. Appl. Sci. Environ. Manage., 12(4), 99-102. Retrieved from: http://www.ajol.info/index.php/jasem/article/view/55247, 2008.
[13] Kovendan, K., Murugan, K., Vincent, S., & Kamalakannan, S. Larvicidal efficacy of Jatropha curcas and bacterial insecticide, Bacillus thuringiensis, against lymphatic filarial vector, Culex quinquefasciatus Say (Diptera:
[14] Culicidae). Parasitology Research, 109, 1251-1257. doi: 10.1007/s00436-011-2368-6, 2011.
[15] Handral, H. K., Pandith, A., and Sd, S. A review on Murraya koenigii multipotential medicinal plant. Asian Journal of Pharmaceutical and Clinical Research, 5(4), 5-14. Retrieved from: http://www.ajpcr.com/Vol5Suppl4/1356.pdf, 2012.
[16] Patil, S., Patil, C., Salunkhe, R., & Salunke, B. Larvicidal activities of six plants extracts against two mosquito species, Aedes aegypti and Anopheles stephensi. Tropical Biomedicine, 27(3), 360-365. Retrieved from: http://www.ncbi.nlm.nih.gov/pubmed/21399575, 2010.
[17] Harve, G., & Kamath, V. Larvicidal activity of plant extracts used alone and in combination with known synthetic larvicidal agents against Aedes aegypti. Indian Journal of Experimental Biology, 42, 1216-1219. Retrieved from: http://www.ncbi.nlm.nih.gov/pubmed/15623234, 2004.
[18] Kaushik, R., and Saini, P. Screening of some semi-arid region plants for larvicidal activity against Aedes aegypti mosquitoes. Journal of Vector-Borne Diseases, 46, 244-246. Retrieved from: http://www.ncbi.nlm.nih.gov/pubmed/19724091, 2009.
[19] Suryawanshi, R., Patil, C., Borase, H., Narkhede, C., and Patil, S. Screening of Rubiaceae and Apocynaceae extracts for mosquito larvicidal potential. Natural Products Reseach, 29 (4) 1-8. doi: 10.1080/14786419.2014.941362, 2014.
[20] Herath, H., & de Silva, S. New Constituents from Gliricidia sepium. Fitoterpia, 71 (6), 722-724. Retrieved. from: http://www.ncbi.nlm.nih.gov/pubmed/11077187, 2000.
[21] World Health Organization. Guidelines for Laboratory and Field Testing of Mosquito Larvicides. World Health Organization Press. Retrived from: http://whqlibdoc.who.int/hq/2005/who_cds_whopes_gcdpp_2005.13.pd,2005.
[22] Kaushik, R., and Saini, P. Screening of some semi-arid region plants for larvicidal activity against Aedes aegypti mosquitoes. Journal of Vector-Borne Diseases, 46, 244-246. Retrieved from: http://www.ncbi.nlm.nih.gov/pubmed/19724091, 2009.
[23] Liu, Z. L., Liuo, Q. Z., Du. S. S., and Deng. Z. W. Mosquito larvicidal activity of alkaloids and limonoids derived from Evodia rutaecarpa unripe fruits against Aedes albopictus (Diptera: Culicidae). Parasitology Research, 111(3), 991-996. doi: 10.1007/s00436-012-2923-9, 2012.
[24] da Silva, H. H. G., da Silva, I. G., dos Santos, R. M. G., Rodrigues, E. R., and Elias, C. N. Larvicidal activity of tannins isolated of Magonia pubescens St. Hil. (Sapindaceae) against Aedes aegypti (Diptera, Culicidae). Revista da Sociedade Brasileira de Medicina Tropical, 396-399. Retrieved from: http://www.scielo.br/pdf/rsbmt/v37n5/21338.pdf, 2004.
[25] Ranganatha, V.L., Begum, A.B., Prashanth, T., Gurupadaswamy, H.D., Madhu, S. K., Shivakumar, S., & Khanum, S. A. Synthesis and larvicidal properties of benzophenone comprise indole analogues against Culex quinquefasciatus. Drug Intervention Today, 5(4), 1-6. doi: 10.1016/j.dit.2013.10.001, 2013.
[26] National Pesticide Center. Capsaicin technical fact sheet. Retrieved rom: http://npic.orst.edu/factsheets/Capsaicintech.pdf.
[1] Tayag, E. A., Roque, Jr., V. G., and Elfa, D. C. Disease surveillance report. Retrieved from: www.doh.gov.ph/sites/default/files/leptoup21.pdf, 201.
[2] World Health Organization. Dengue: Guidelines for diagnosis, treatment, prevention and control. Retrieved from: www.who.int/tdr/publications/documents/dengue-diagnosis.pdf, 2009.
[3] Negrelle, R. R. B., and Gomes, E. C. Cymbopogon citratus (DC.) Stapf: Chemical composition and biological activities. Rev. Bras. Pt. Med., 9(1), 80-92. Retrieved from: http://www.sbpmed.org.br/download/issn_07_1/revisao_v9_n1.pd,2007.
[4] Karunamoorthi, K., and Ilango, K. Larvicidal activity of Cymbopogon citratus (DC Stapf. and Croton macrostachyus Del. against Anopheles arabiensis Patton, a potent malarial vector. European Review for Medical and Pharmacological Sciences, 14, 57-62. Retrieved from: http://www.ncbi.nlm.nih.gov/pubmed/20184090, 2007.
[5] Cavalcanti, E., de Morais, S., Lima, M. A., & Santana, E. Larvicidal Activity of Essential Oils from Brazilian Plants against Aedes aegypti L. Mem. Inst. Oswaldo Cruz, 99(5), 541-544. Retrieved from: http://www.ncbi.nlm.nih.gov/pubmed/15543421, 2004.
[6] Quisumbing, E. Medicinal Plants of the Philippines. Quezon City, Philippines: JMC Press, Inc., 1978.
[7] Madhumathy, A., Aivazi, A. A., and Vijayan, V. Larvicidal efficacy of Capsicum annum against Anopheles stephensi and Culex quinquefasciatus. Journal of Vecotr Borne Diseases, 44, 223-226. Retrieved from: http://www.ncbi.nlm.nih.gov/pubmed/17896626, 2007.
[8] Vinayaka, K. S., Prakish Kekuda, T. R. and Nandini, K.C. Potent insecticidal activity of fruits and leaves of Capsicum frutescens (L.) var. longa (Solanaceae). Der Pharmacia Lettre, 2(4), 172-176. Retrieved from: http://scholarsresearchlibrary.com/DPL-vol2-iss4/DPL-2010-2-4-172-176.pdf, 2010.
[9] Batish, D. R., Singh, H. P., Kohli, R. K., & Kaur, S. Eucalyptus essential oil as a natu ral pesticide. Forest Ecology and Management, 256, 2166-2174. doi: 10.1016/j.foreco.2008.08.008, 2008.
[10] Kumar, S., Malhotra, R., and Kumar, D. Euphorbia hirta: Its chemistry, traditional and medicinal uses, and pharmacological activities. Pharmacgon. Rev., 4(7), 58-61. doi: 10.4103/0973-7847.65327, 2010.
[11] Patil, S., Patil, C., Salunkhe, R., & Salunke, B. Larvicidal activities of six plants extracts against two mosquito species, Aedes aegypti and Anopheles stephensi. Tropical Biomedicine, 27(3), 360-365. Retrieved from: http://www.ncbi.nlm.nih.gov/pubmed/21399575, 2010.
[12] Uche, F., and Aprioku, J. The Phytochemical Constituents, Analgesic and Anti-inflammatory effects of methanol extract of Jatropha curcas leaves in Mice and Wister albino rats. J. Appl. Sci. Environ. Manage., 12(4), 99-102. Retrieved from: http://www.ajol.info/index.php/jasem/article/view/55247, 2008.
[13] Kovendan, K., Murugan, K., Vincent, S., & Kamalakannan, S. Larvicidal efficacy of Jatropha curcas and bacterial insecticide, Bacillus thuringiensis, against lymphatic filarial vector, Culex quinquefasciatus Say (Diptera:
[14] Culicidae). Parasitology Research, 109, 1251-1257. doi: 10.1007/s00436-011-2368-6, 2011.
[15] Handral, H. K., Pandith, A., and Sd, S. A review on Murraya koenigii multipotential medicinal plant. Asian Journal of Pharmaceutical and Clinical Research, 5(4), 5-14. Retrieved from: http://www.ajpcr.com/Vol5Suppl4/1356.pdf, 2012.
[16] Patil, S., Patil, C., Salunkhe, R., & Salunke, B. Larvicidal activities of six plants extracts against two mosquito species, Aedes aegypti and Anopheles stephensi. Tropical Biomedicine, 27(3), 360-365. Retrieved from: http://www.ncbi.nlm.nih.gov/pubmed/21399575, 2010.
[17] Harve, G., & Kamath, V. Larvicidal activity of plant extracts used alone and in combination with known synthetic larvicidal agents against Aedes aegypti. Indian Journal of Experimental Biology, 42, 1216-1219. Retrieved from: http://www.ncbi.nlm.nih.gov/pubmed/15623234, 2004.
[18] Kaushik, R., and Saini, P. Screening of some semi-arid region plants for larvicidal activity against Aedes aegypti mosquitoes. Journal of Vector-Borne Diseases, 46, 244-246. Retrieved from: http://www.ncbi.nlm.nih.gov/pubmed/19724091, 2009.
[19] Suryawanshi, R., Patil, C., Borase, H., Narkhede, C., and Patil, S. Screening of Rubiaceae and Apocynaceae extracts for mosquito larvicidal potential. Natural Products Reseach, 29 (4) 1-8. doi: 10.1080/14786419.2014.941362, 2014.
[20] Herath, H., & de Silva, S. New Constituents from Gliricidia sepium. Fitoterpia, 71 (6), 722-724. Retrieved. from: http://www.ncbi.nlm.nih.gov/pubmed/11077187, 2000.
[21] World Health Organization. Guidelines for Laboratory and Field Testing of Mosquito Larvicides. World Health Organization Press. Retrived from: http://whqlibdoc.who.int/hq/2005/who_cds_whopes_gcdpp_2005.13.pd,2005.
[22] Kaushik, R., and Saini, P. Screening of some semi-arid region plants for larvicidal activity against Aedes aegypti mosquitoes. Journal of Vector-Borne Diseases, 46, 244-246. Retrieved from: http://www.ncbi.nlm.nih.gov/pubmed/19724091, 2009.
[23] Liu, Z. L., Liuo, Q. Z., Du. S. S., and Deng. Z. W. Mosquito larvicidal activity of alkaloids and limonoids derived from Evodia rutaecarpa unripe fruits against Aedes albopictus (Diptera: Culicidae). Parasitology Research, 111(3), 991-996. doi: 10.1007/s00436-012-2923-9, 2012.
[24] da Silva, H. H. G., da Silva, I. G., dos Santos, R. M. G., Rodrigues, E. R., and Elias, C. N. Larvicidal activity of tannins isolated of Magonia pubescens St. Hil. (Sapindaceae) against Aedes aegypti (Diptera, Culicidae). Revista da Sociedade Brasileira de Medicina Tropical, 396-399. Retrieved from: http://www.scielo.br/pdf/rsbmt/v37n5/21338.pdf, 2004.
[25] Ranganatha, V.L., Begum, A.B., Prashanth, T., Gurupadaswamy, H.D., Madhu, S. K., Shivakumar, S., & Khanum, S. A. Synthesis and larvicidal properties of benzophenone comprise indole analogues against Culex quinquefasciatus. Drug Intervention Today, 5(4), 1-6. doi: 10.1016/j.dit.2013.10.001, 2013.
[26] National Pesticide Center. Capsaicin technical fact sheet. Retrieved rom: http://npic.orst.edu/factsheets/Capsaicintech.pdf.
@article{"International Journal of Biological, Life and Agricultural Sciences:75242", author = "Michael Russelle S. Alvarez and Noel S. Quiming and Francisco M. Heralde", title = "Screening for Larvicidal Activity of Aqueous and Ethanolic Extracts of Fourteen Selected Plants and Formulation of a Larvicide against Aedes aegypti (Linn.) and Aedes albopictus (Skuse) Larvae", abstract = "This study aims to: a) obtain ethanolic (95% EtOH) and aqueous extracts of Selaginella elmeri, Christella dentata, Elatostema sinnatum, Curculigo capitulata, Euphorbia hirta, Murraya koenigii, Alpinia speciosa, Cymbopogon citratus, Eucalyptus globulus, Jatropha curcas, Psidium guajava, Gliricidia sepium, Ixora coccinea and Capsicum frutescens and screen them for larvicidal activities against Aedes aegypti (Linn.) and Aedes albopictus (Skuse) larvae; b) to fractionate the most active extract and determine the most active fraction; c) to determine the larvicidal properties of the most active extract and fraction against by computing their percentage mortality, LC50, and LC90 after 24 and 48 hours of exposure; and d) to determine the nature of the components of the active extracts and fractions using phytochemical screening. Ethanolic (95% EtOH) and aqueous extracts of the selected plants will be screened for potential larvicidal activity against Ae. aegypti and Ae. albopictus using standard procedures and 1% malathion and a Piper nigrum based ovicide-larvicide by the Department of Science and Technology as positive controls. The results were analyzed using One-Way ANOVA with Tukey’s and Dunnett’s test. The most active extract will be subjected to partial fractionation using normal-phase column chromatography, and the fractions subsequently screened to determine the most active fraction. The most active extract and fraction were subjected to dose-response assay and probit analysis to determine the LC50 and LC90 after 24 and 48 hours of exposure. The active extracts and fractions will be screened for phytochemical content. The ethanolic extracts of C. citratus, E. hirta, I. coccinea, G. sepium, M. koenigii, E globulus, J. curcas and C. frutescens exhibited significant larvicidal activity, with C. frutescens being the most active. After fractionation, the ethyl acetate fraction was found to be the most active. Phytochemical screening of the extracts revealed the presence of alkaloids, tannins, indoles and steroids. A formulation using talcum powder–300 mg fraction per 1 g talcum powder–was made and again tested for larvicidal activity. At 2 g/L, the formulation proved effective in killing all of the test larvae after 24 hours.
", keywords = "Larvicidal activity screening, partial purification, dose-response assay, Capsicum frutescens.", volume = "9", number = "11", pages = "1183-7", }
