J Arthropod-Borne Dis, December 2020, 14(4): 391–399 S Sayono et al.: Larvicidal Activity of … 391 http://jad.tums.ac.ir Published Online: December 31, 2020 Original Article Larvicidal Activity of Ethyl Acetate Extract of Derris elliptica Root against the Third-Instar Larvae of Cypermethrin-Resistant Aedes aegypti Offspring *Sayono Sayono1; Risyandi Anwar2; Didik Sumanto3 1Faculty of Public Health, Universitas Muhammadiyah Semarang, Semarang, Indonesia 2Herbal medicine research of Dentistry Faculty, Universitas Muhammadiyah Semarang, Semarang, Indonesia 3Laboratory of Epidemiology and Tropical Diseases, Faculty of Public Health, Universitas Muhammadiyah Semarang, Semarang, Indonesia *Corresponding author: Dr Sayono Sayono, E-mail: say.epid@gmail.com (Received 30 Nov 2019; accepted 15 Dec 2020) Abstract Background: Derris elliptica extracts have a high larvicidal potential against the laboratory strain of Aedes aegypti larvae, but the effect on offspring larvae of pyrethroid-resistant strains of the species is lack understood. This study aimed to determine the larvicidal activity of the ethyl acetate extract of tuba root against the third-instar larvae of the Cypermethrin-resistant Ae. aegypti offspring. Methods: The experimental study occupied four levels of ethyl acetate extract of D. elliptica namely 10, 25, 50, and 100 ppm, and each level was four times replicated. As many as twenty of healthy third-instar larvae, offspring of Cy- permethrin-resistant Ae. aegypti were subjected to each experiment group. Larval mortality rate and lethal concentration 50% subject (LC50) were calculated after 24 and 48 hours of exposure time. Results: Mortality of larvae increased directly proportional to the increase of extract concentration. Larval mortality rates after 24 and 48 hours of exposure were 40–67.5% and 62.5–97.5%, and LC50 were 34.945 and 6.461ppm, respectively. Conclusion: The ethyl acetate extract of D. elliptica has the high effectiveness larvicidal potential against the third- instar larvae, offspring of the Cypermethrin-resistant Ae. aegypti. Isolation of the specific compound is necessarily done to obtain the active ingredient for larvicide formulation. Keywords: Larvicidal activity; Ethyl acetate extract; Derris elliptica root; Cypermethrin resistant; Aedes aegypti Introduction The resistance of Ae. aegypti to several py- rethroid and organophosphate insecticide com- pounds such as deltamethrin, lambda cyhalo- thrin, cypermethrin, malathion, and temephos (1, 2) inhibits the public health action in erad- icating the dengue vector, and intrigues research- ers to find the other active ingredients as the al- ternatives. Natural chemical compounds (3), in- cluding D. elliptica roots (4), are interesting to study for several reasons including but not lim- ited to readily degraded and there is no bioac- cumulation in the environment (5). Research- ers have proven that D. elliptica extracts have high larvicidal potential against the laboratory strain larvae of Ae. aegypti (3, 4, 6, 7). Howev- er, when methanol extract of D. elliptica was exposed to the filed-caught larvae of Ae. ae- gypti showed a lower larvicidal potential (8). This fact showed that the different extract types of the tubal root have different effects against the different strains of Ae. aegypti larvae where the field-caught larvae were more resistant to the phytochemical compound. The results of monitoring of dengue vector susceptibility in Central Java, Indonesia showed a wide spread of resistance to cypermethrin 0.05 % (1), as occurs in various Dengue endemic ar- eas in other countries (4, 9, 10). Cypermethrin is one of the active ingredients of pyrethroid class insecticide which has caused knockdown resistance (kdr) (11). This resistance mechanism was indicated with the target site insensitivity Copyright © 2020 Iranian Scientific Society of Biology & Control of Diseases Vectors, and Tehran University of Medical Sciences. Published by Tehran University of Medical Sciences. This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International license (https://creativecommons.org/licenses/by-nc/4.0/). Non-commercial uses of the work are permitted, provided the original work is properly cited. http://jad.tums.ac.ir/ https://creativecommons.org/licenses/by-nc/4.0/ J Arthropod-Borne Dis, December 2020, 14(4): 391–399 S Sayono et al.: Larvicidal Activity of … 392 http://jad.tums.ac.ir Published Online: December 31, 2020 in the voltage-gated sodium channel (VGSC) gene (1, 12). The action mechanism of cyper- methrin is different from the temephos. This compound is an active ingredient of organo- phosphate insecticide class which inhibited the acetylcholinesterase enzyme (13). This differ- ent mechanism of action is interesting to be studied in understanding the larvicidal activity spectrum of D. elliptica root extracts. The main biochemical compounds of D. el- liptica are alkaloids, flavonoids, sterols, tannins, and triterpenoids (14, 15), and rotenone is the most important of a specific compound of fla- vonoid (16). These compounds have a toxic ef- fect that kills insect larvae through disrupting mechanisms of the endocrine and hormonal sys- tems (14) and reducing the esterase and monoox- ygenase enzymes (16). Initial studies showed that ethyl acetate, methanol, and n-hexane ex- tracts of D. elliptica that have different polar- ity effectively killed Ae. aegypti larvae which were susceptible to temephos 0.02ppm (17), but on the other hand, the ethanol extract type has a lower effect against the temephos-re- sistant strains (18). The lethal effects of dif- ferent specific phytochemicals contained in D. elliptica root extracts against the offspring lar- vae of the cypermethrin-resistant strain Ae. aegypti is still lack understood and is interest- ing to be studied. It is important to evaluate the larvicidal effect of the semi polar extract, ethyl acetate against this strain. This study aimed to determine the larvicidal activity of ethyl ace- tate extract of D. elliptica root against the third- instar larvae of the cypermethrin-resistant Ae. aegypti offspring. Materials and Methods This experiment is the early part of an on- going study ‘Isolation of specific compounds of Derris elliptica as the larvicidal ingredients against Aedes aegypti mosquito in the dengue control’. Ethyl acetate extract is the last step of the sequential extraction process (19, 20). In the summary modification, the extraction pro- cess started by maceration of the tuba root pow- der in methanol solvent for 3 x 24 hours, and then filtered. The clean part of the liquid is evaporated and produced the methanol extract (the crude extract). Furthermore, the crude ex- tract was partitioned liquid-liquid with n-hex- ane solvent to bind the nonpolar lead com- pounds and resulted in water fraction and n- hexane fraction. The water fraction obtained was partitioned with ethyl acetate to bind the semi-polar lead compounds and produced the water fraction and ethyl acetate fraction. All fractions produced were evaporated by using a rotary evaporator to produce four types of ex- tracts, including the ethyl acetate extract which was first completed. The subjects of this study were the offspring filial 2 (F2) larvae of the cypermethrin 0.05% resistant strain of Ae. aegypti. The parental mos- quitoes were the F1 larvae of the Ae. aegypti that is reared from F0 larvae obtaining from a household survey in the Dengue endemic ar- eas in the Community Health Center of Kedung Mundu, Tembalang District, Semarang City, and subjected to bioassay test using the Cy- permetrhin-0.05% compound. The result of the bioassay test showed a mortality rate of 85%, which indicated that the mosquito population was resistant to the pyrethroid compound (21). Larvae were maintained in the Epidemiology and Tropical Diseases laboratory of Public Health Faculty of Universitas Muhammadiyah Semarang, Indonesia. The larvae were placed on a plastic tray containing tap water. Condi- tions of temperature and humidity were main- tained in the range of 28±2 °C and 75±10%. The larvae were fed dog food. Bioassay tests apply the WHO standard procedures for larvi- cide testing (21). There are three important parts at this stage, namely preparation of extract con- centration, selection of research subjects, and exposure of research subjects with various D. elliptica extracts. The concentration of the bi- oassay test used a range of 10, 25, 50, and 100 ppm, the effective concentration in another study using Ae. aegypti larvae of laboratory http://jad.tums.ac.ir/ J Arthropod-Borne Dis, December 2020, 14(4): 391–399 S Sayono et al.: Larvicidal Activity of … 393 http://jad.tums.ac.ir Published Online: December 31, 2020 strains (17). The subject of the research was the third instar larvae offspring of the Cy- permethrin-resistant Ae. aegypti, in intact con- dition, and actively moving. As many as twenty larvae were subjected to each experiment group. Two control groups, negative (tap water) and positive (temephos 0.02ppm) control were fol- lowed. The effectiveness of the larvicidal ac- tivity of the ethyl acetate extract of D. ellip- tica root was determined by the LC50 that was obtained from probit analysis. This LC50 will be compared with the LC50 of previous experiment results of the same extract type against the la- boratory (susceptible) strain of Ae. aegypti lar- vae (17). Analysis data was performed descrip- tively and analytically by using SPSS statisti- cal software version 15.0. The research protocol obtained ethical approval from the Ethics Com- mittee of Health Research of Public Health Fac- ulty of Universitas Muhammadiyah Semarang with registration number 231/KEPK-FKM/ UNIMUS/2019. Results The ethyl acetic extracts of D. elliptica root showed the larvicidal activity against the Ae. aegypti larvae of offspring from the resistant parental to cypermethrin adulticide. There was an increase in the larval mortality rate of Ae. aegypti larvae after 24 and 48 hours of expo- sure to the ethyl acetic extract from 40–67.5% to 62.5–97.5% (Table 1), with LC50 of 34.945 and 6.461ppm, respectively (Table 2). The lar- val mortality rate increased directly with the ex- tract concentration. There were no larvae died in the negative control, and 100% of larvae died in the positive control. The trend of the knock- down larvae showed that the slow larvicidal ac- tivity of the ethyl acetate extract of D. elliptica root to the third-instar larvae, offspring F2 of cypermethrin-resistant Ae. aegypti (Fig. 1). Sta- tistical analysis showed the differences in lar- val mortality rate based on the dosage and ex- posure time (Fig. 2). Fig. 1. The trend of larval knockdown rate in each extracts concentration based on the exposure time. The colored-line represents the concentrations of the extract http://jad.tums.ac.ir/ J Arthropod-Borne Dis, December 2020, 14(4): 391–399 S Sayono et al.: Larvicidal Activity of … 394 http://jad.tums.ac.ir Published Online: December 31, 2020 Table 1. Larvicidal activity of the ethyl acetate extract type of Derris elliptica against the third-instar larvae of cypermethrin-resistant Aedes aegypti offspring Concentration (ppm) 24-hours mortality rate (%) 48-hours mortality rate (%) Min Max Mean Min Max Mean 0 (dw) 0 0 0 0 0 0 10 30 40 35 60 65 62.5 25 40 40 40 80 90 85 50 40 70 55 90 100 95 100 65 70 67.5 95 100 97.5 0.02 (tem) 100 100 100 100 100 100 dw= distilled water; tem= temephos Table 2. The LC50 of larvicidal activity of the Derris elliptica ethyl acetate extract against offspring larvae of the cypermethrin-resistant Aedes aegypti Exposure time (hours) Regression equation LC50 (95% confidence limits) 24 Y= -1.331+0.862X 34.945 (18.179–70.780) 48 Y= -1.419+1.751X 6.461 (2.206–10.359) 35 40 55 67.5 62.5 85 95 97.5 0 20 40 60 80 100 120 10 25 50 100 M o r ta li ty ( % ) Dosage (ppm) 24h 48h Fig. 2. Mortality rate comparison of Aedes aegypti larvae after 24h and 48h exposure of ethyl acetate extract of Derris elliptica root. The differences of larval mortality rate are indicated by the letters a, b and c Discussion Results of the experiment showed that the ethyl acetic extract of D. ellitpica has a high larvicidal activity against the third-instar lar- vae of the cypermethrin-resistant Ae. aegypti F2 offspring, although at the concentration of 100ppm for 24 hours, the larvicidal effect of the ethyl acetate extract type is still lower than the larvicidal activity of temephos 0.02ppm. Alt- hough the ethyl acetate extract type of tuba root shows lower larvicidal activity than temephos, this extract still indicates high larvicidal poten- tial because its LC50 is lower than 50ppm. A previous study reported that the effective lar- vicidal activity of plant extracts was categorized into three levels, namely high (LC50< 50ppm), moderate (LC50< 100ppm), and low LC50< 750 ppm) (4). The results of this experiment indi- cate that the potency of the ethyl acetate extract http://jad.tums.ac.ir/ J Arthropod-Borne Dis, December 2020, 14(4): 391–399 S Sayono et al.: Larvicidal Activity of … 395 http://jad.tums.ac.ir Published Online: December 31, 2020 of D. elliptica root against the third-instar lar- vae of cypermethrin-resistant Ae. aegypti F2 offspring is 84.4% lower than the same extract potency against the susceptible (laboratory) strain (17) indicating by the LC50 bioassay test for 24 hours, respectively 34,945ppm and 21,063 ppm. This extract also had a higher larvicidal potential against F2 offspring larvae of Cyper- methrin-resistant Ae. aegypti rather than the temephos-resistant offspring (18). The larvicidal activity of this extract was also better than the methanolic extract of M. glaziovii peel (22), A. pinata (23), T. patula (24), H. forskaolii (25), O. campechianum and O. quixos (26), and A. oc- cidentale (27) against the third-instar larvae of Ae. aegypti, although lower than the specific isolate compound of F. vulgare (28) and P. aduncum (26) essential oils, and P. foetida ethyl acetate extract (29). These preliminary data and information have given new hope that this ex- tract can be an alternative ingredient of larvi- cide to inhibit the growth of Ae. aegypti larvae, even to the strains that are already resistant to cypermethrin adulticide. These results also indicate that temephos is still effective in killing the third-instar larvae offspring F2 of the cypermethrin-resistant Ae. aegypti. This condition showed that there is still a way to eradicate the Dengue vector, even from strains that have been resistant to other insecti- cides because each insecticide compound has a different mode of action (30, 31). Cyperme- thrin is a compound of the pyrethroid class, an insecticide group that disrupts the function of sodium channels in insect nerves (32). Under normal conditions, the voltage-gated sodium channel (VGSC) gene works ‘open’ and ‘close’ to regulate electrical impulses into the cell. The linkage of the pyrethroid insecticide molecule to the gene disrupts the nerve regulation and impulse of the nerve flowing continuously so that the insects become convulsion and died (33). However, if the point mutations occur in this gene, the linkage of the pyrethroid insec- ticide molecule does not affect the life of the insect, and this condition caused the kdr (34). The high larvicidal activity of the ethyl ace- tic extract of D. elliptica root against the third instar larvae, the offspring of the cypermethrin- resistant Ae. aegypti indicates that this extract has a different mechanism of action than adul- ticide cypermethrin. The D. elliptica extract contains several lead compounds such as tan- nins, phlobatannins, terpenoids, cardiac gly- cosides, and flavonoids (35) mainly rotenone and rotenoids (36). Mode of action of rotenone is inhibition the cellular respiration, while pyre- thrins the active compound of the pyrethroid insecticide has a mode of action in disruption of the sodium and potassium ions exchange (37). It means that the exploration of specific isolates of D. elliptica extract has the oppor- tunity to be developed into larvicidal bioactive compounds with different modes of action. On the other hand, the effectiveness of temephos larvicide in killing the offspring larvae of the cypermethrin resistant strain of Ae. aegypti proved that rotational insecticide with differ- ent modes of action and target sites is neces- sary done. Temephos is a compound that plays a role in protein carbonylation so that it causes the general oxidative damage in larval cellular of insects (38). The maximum effect of the ethyl acetate ex- tract of D. elliptica was achieved at 48 hours of exposure time. This condition indicated that the mode of action of this extract is slower than temephos. It can be understood that temephos is a pure chemical compound, while the plant extract still contains many chemical compounds, which may have antagonistic effects (39). Ex- traction with ethyl acetate solvent has selected chemical compounds that are semi-polar, ac- cording to the nature of the solvent. However, the extraction results still allow the dissolu- tion of many plant chemical compounds with various modes of action although flavonoid was the dominant compound (40). Therefore, the pure compounds from these extracts that have the best larvicidal activity are necessari- ly understood. http://jad.tums.ac.ir/ J Arthropod-Borne Dis, December 2020, 14(4): 391–399 S Sayono et al.: Larvicidal Activity of … 396 http://jad.tums.ac.ir Published Online: December 31, 2020 Conclusion The ethyl acetate crude extract of D. ellip- tica root has a high larvicidal activity against the third-instar larvae, offspring of the cyper- methrin-resistant strain of Ae. aegypti, although the effect is lower and slower than temephos. Isolation of the pure compounds of the extract is needed to find the specific active compounds for larvicide formulation. Acknowledgements The authors wish to thank President of Uni- versitas Muhammadiyah Semarang for the re- search permission for experimenting in labor- atory study at the Epidemiology and Tropical Diseases laboratory; Dean of Mathematical and Natural Sciences Faculty of Universitas Garut, West Java, Indonesia; Directorate General of Research and Development Strengthening, Min- istry of Research, Technology and Higher Ed- ucation for the funding of the study. References 1. Sayono S, Hidayati APN, Fahri S, Suman- to D, Dharmana E, Hadisaputro S, Asih PBS, Syafruddin D (2016) Distribution of Voltage-Gated Sodium Channel (Nav) alleles among the Aedes aegypti popula- tions in Central Java Province and its as- sociation with resistance to pyrethroid in- secticides. PLoS One. 11(3): e0150577. 2. Araújo AP, Paiva MHS, Cabral AM, Cav- alcanti AEHD, Pessoa LFF, Diniz DFA, Helvecio E, Silva EVG, Silva NM, Anastácio DB, Pontes C, Nunes V, Souza MFM, Magalhães FJR, Santos MAVM, Ayres CFJ (2019) Screening Aedes aegypti (Diptera: Culicidae) pop- ulations from Pernambuco, Brazil for resistance to Temephos, Diflubenzuron, and Cypermethrin and characterization of potential resistance mechanisms. J In- sect Sci. 19(3): 16. 3. Dohutia C, Bhattacharyya DR, Sharma SK, Mohapatra PK, Bhattacharjee K, Gogoi K, Gogoi P, Mahanta J, Prakash A (2015) Larvicidal activity of few select indige- nous plants of North East India against disease vector mosquitoes (Diptera: Cu- licidae). Trop Biomed. 32(1): 17–23. 4. Komalamisra N, Trongtokit Y, Rongsriyam R, Apiwathnasorn C (2005) Screening for larvacidal activity in some Thai plants against four mosquito vector species. Southeast Asian J Trop Med Public Health. 36(6): 1412–1422. 5. Arnason JT, Sims SR, Scott IM (2012) Natu- ral products from plants as insecticides. Encyclopedia of Life Support System (EoLSS). Phytochemistry and pharma- cognosy. Available at: http://www.eolss.net/sample- chapters/c06/e6-151-13.pdf 6. Zubairi SI, Sarmidi MR, Aziz RA (2015) A preliminary study on mosquito larvi- cidal efficacy of rotenone extracted from Malaysia Derris sp. J Teknologi. 76(1): 275–279. 7. Komansilan A, Suriani NW, Lawalata H (2017) Test toxic tuba root extract as a natural insecticide on larvae of Aedes aegypti mosquito vector of dengue fe- ver. Int J Chemtech Res. 10(4): 522–528. 8. Sayono S, Nurullita U, Suryani M (2010) Pengaruh konsentrasi flavonoid dalam ekstrak akar tuba (Derris elliptica) ter- hadap kematian larva Aedes aegypti. [In Indonesian]. J Kesehat Masy Indones. 6 (1): 38–47. 9. Sayono S, Nurullita U (2016) Situasi terkini vektor dengue (Aedes aegypti) di Jawa Tengah, Indonesian. Kemas. Jurnal Kesehatan Masyarakat. 11(2): 96–105. 10. Moyes C, Vontas J, Martins AJ, Ng LC, Koou SY, Dusfour I, Raghavendra K, Pinto J, Corbel V, David JP, Weetman D (2017) Contemporary status of insec- ticides resistance in the major Aedes vectors of arboviruses infecting humans. http://jad.tums.ac.ir/ http://www.eolss.net/sample-chapters/c06/e6-151-13.pdf http://www.eolss.net/sample-chapters/c06/e6-151-13.pdf J Arthropod-Borne Dis, December 2020, 14(4): 391–399 S Sayono et al.: Larvicidal Activity of … 397 http://jad.tums.ac.ir Published Online: December 31, 2020 PLoS Negl Trop Dis. 11(7): e0005625. 11. Singh AK, Tiwari MN, Prakash O, Sing MP (2012) A current review of Cyper- methrin-induced neurotoxicity and nigro- striatal dopaminergic neurodegeneration. Curr Neuropharmacol. 10(1): 64–71. 12. Kuswah RBS, Kaur T, Dykes CL, Kumar HR, Kapoor N, Sing OP (2020) A new knockdown resistance (kdr) mutation, F 1534L, in the Voltage-gated Sodium Channer of Aedes aegypti, co-occurring with F1534C, S989P, and V1016G. Par- asit Vectors. 13: 327. 13. Insecticide Resistance Action Committee (IRAC) (2019) IRAC mode of action classification scheme. IRAC Internation- al Working Group. pp. 5–7. 14. Ge Y, Liu P, Yang R, Zhang L, Chen H, Camara I, Liu Y, Shi W (2015) Insecti- cidal constituents and activity of alkaloids from Cynanchum mongolicum. Molecules. 20: 17483–17492. 15. Khan MR, Omoloso AD, Barewai Y (2006) Antimicrobial activity of the Derris elliptica, Derris indica and Derris trifo- liata extractives. Fitoterapia. 77(4): 327– 330. 16. Visetson S, Milne M (2001) Effect of root extract from Derris (Derris elliptica Benth) on mortality and detoxification enzyme levels in the Demondback Moth larvae (Plutella xylostella Linn.). Kaset- sart J (Nat. Sci.). 35: 157–163. 17. Sayono S, Anwar R, Sumanto D (2020) Evaluation of toxicity in four extract types of Tuba root against Dengue vec- tor, Aedes aegypti (Diptera: Cullicidae) larvae. Pak J Biol Sci. 23(12): 1530– 1538. 18. Sayono S, Permatasari A, Sumanto D (2019) The effectiveness of Derris el- liptica (Wall.) Benth root extract against Temephos-resistant Aedes aegypti larvae. IOP Conference Series: Earth and Envi- ronmental Science. 292. The 1st Interna- tional Conference on Food Science and Technology 28–29 November 2018, Uni- versitas Muhammadiyah Semarang, Se- marang, Indonesia, p. 102052. 19. Liu Z (2008) Preparation of botanical sam- ples for biomedical research. Endocr Metab Immune Disord Drug Targets. 8 (2): 112–121. 20. Li XJ, Hareyama T, Tezuka Y, Zhang Y, Miyahara T, Kadota S (2005) Five new oleanolic acid glycosides from Achy- ranthes bidentata with inhibitory activ- ity on osteoclast formation. Planta Med. 71(7): 673–679. 21. World Health Organization (2016) Monitoring and managing insecticide resistance in Aedes mosquito popula- tions. Interim guidance for entomolo- gists. Available at: file:///C:/Users/032137~1/AppData/Lo cal/Temp/WHO_ZIKV_VC_16.1_eng .pdf 22. Sayono S, Safira FA, Anwar R (2019) In-vitro study on the larvicidal activity of Manihot glaziovii peel extract against Aedes aegypti larvae. Ann Parasitol. 65 (4): 403–410. 23. Ravi R, Zulkrnin NSH, Rozhan NN, Nik Yusoff NR, Mat Rasat MS, Ahmad MI, Ishak IH, Mohd Amin MF (2018) Chem- ical composition and larvicidal activities of Azolla pinnata extracts against Aedes (Diptera: Culicidae). PLoS One. 13(11): e0206982. 24. Krzyzaniak LM, Antonelli-Ushirobira TM, Panizzon G, Luiza Sereia AL, Souza JRP, Zequi JAC, Novello CR, Lopes GC, Medeiros DC, Silva DB, Leite-Mello EVS, Mello JCP (2017) Larvicidal ac- tivity against Aedes aegypti and chemi- cal characterization of the inflorescences of Tagetes patula. Evid Based Comple- ment Alternat Med. 2017: 9602368. 25. Sillo AJ, Makirita WE, Swai H, Chacha M (2019) Larvicidal activity of Hypoestes forskaolii (Vahl) R.Br root extracts against Anopheles gambiae Giless.s, Aedes ae- http://jad.tums.ac.ir/ file:///C:/Users/0321378490/AppData/Local/Temp/WHO_ZIKV_VC_16.1_eng.pdf file:///C:/Users/0321378490/AppData/Local/Temp/WHO_ZIKV_VC_16.1_eng.pdf file:///C:/Users/0321378490/AppData/Local/Temp/WHO_ZIKV_VC_16.1_eng.pdf https://www.ncbi.nlm.nih.gov/pubmed/?term=Amin%20MF%5BAuthor%5D&cauthor=true&cauthor_uid=30399167 J Arthropod-Borne Dis, December 2020, 14(4): 391–399 S Sayono et al.: Larvicidal Activity of … 398 http://jad.tums.ac.ir Published Online: December 31, 2020 gypti L, and Culex quinquefasciatus Say. J Exp Pharmacol. 11: 23–27. 26. Scalvenzi L, Radice M, Toma L, Severini F, Boccolini D, Bella A, Guerrini A, Tac- chini M, Sacchetti G, Chiurato M, Romi R, Luca MD (2019) Larvicidal activity of Ocimum campechianum, Ocotea quixos and Piper aduncum essential oils against Aedes aegypti. Parasite. 26: 23. 27. Amado CJRR, Souto CRNP, Magalhães MS, Arranz CJCE, Carvalho CJCT (2017) Chemical composition and larvicidal ac- tivity of cashew nutshell ethanolic ex- tract against mosquito larvae. Rev Cub Quim. 29(3): 330–340. 28. Rocha DK, Matos O, Novo MT, Figueire- do AC, Delgado M, Moiteiro C (2015) Larvicidal activity against Aedes aegypti of Foeniculum vulgare essential oils from Portugal and Cape Verde. Nat Prod Com- mun. 10 (4): 677–682. 29. Maywan H (2016) Aktivitas larvasida ekstrak etanol, fraksi n-heksan, etil asetat, dan metanol daun sembukan terhadap lar- va nyamuk Aedes aegypti dan Anopheles instar III. Trad Med J. 21(3): 137–142. 30. Das SK (2013) Mode of action of pesti- cides and the novel trends-A critical review. Int Res J Agric Sci Soil. 3(11): 393–401. 31. Spark TC, Nauen R (2015) IRAC: Mode of action classification and insecticide resistance management. Pestic Biochem Physiol. 121: 122–128. 32. Soderlund DM (2012) Molecular mecha- nisms of pyrethroid insecticide neuro- toxicity: recent advances. Arch Toxicol. 86(2): 165–181. 33. Silver KS, Du Y, Nomura Y, Olivera EE, Salgado VL, Zhoro BS, Dong K (2014) Voltage-Gated Sodium Channels as in- secticide targets. Adv In Insect Phys. 46: 389–433. 34. Dong K, Du Y, Rinkevich F, Nomura Y, Xu P, Wang L, Silver K, Zhorov BS (2014) Molecular biology of insect so- dium channels and pyrethroid resistance. Insect Biochem Mol Biol. 50: 1–17. 35. Lagunay RAE, Uy MM (2015) Evalua- tion of the phytochemical constituents of the leaves of Ficus minahassae Tesym and De Vr., Casuarina equisetifolia Linn., Leucosyke capitellata (Pior) Wedd., Cas- sia sophera Linn., Derris elliptica Benth., Cyperus brevifolius (Rottb.) Hassk., Piper abbreviatum Opiz., Ixora chinensis Lam., Leea aculeata Blume, and Drymoglos- sum piloselloides Linn. AAB Bioflux. 7 (1): 51–58. 36. Zubairi SI, Ramli KA, Majid FAA, Sarmidi MR, Aziz RA (2005) Biological screen- ing on the extract of Derris elliptica. Pro- ceeding of Kustem 4th Annual Seminar on Sustainability Science and Manage- ment, 2005 May 2–3, Primula Beach Re- sort, Kuala Terengganu. Available at: http://eprints.utm.my/id/eprint/5263/ 37. Ghosh A, Chowdhury N, Chandra G (2012) Plant extracts as potential mosquito lar- vicides. Indian J Med Res. 135: 581–598. 38. Rodríguez-Cavallo E, Guarnizo-Méndez J, Yépez-Terrill A, Cárdenas-Rivero A, Díaz-Castillo F, Méndez-Cuadro D (2018) Protein carbonylation is a mediator in larvicidal mechanisms of Tabernaemon- tana cymosa ethanolic extract. J King Saud Univ Sci. 31(4): 464–471. 39. Millugo TK, Osoma LK, Ochanda JO, Owuor BO, Wamunyokoli FA, Oyugi JO, Ochieng JW (2013) Antagonistic ef- fect of alkaloids and saponins on bioac- tivity in the quinine tree (Rauvolfia caffra- sond.): further evidence to support bio- technology in traditional medicinal plants. BMC Complement Altern Med. 13: 285. 40. Thavamoney N, Sivanadian L, Tee LH, Khoo HE, Prasad KN, Kong KW (2018) Extraction and recovery of phytochemi- cal components and antioxidative prop- erties in fruit parts of Dacryodes ros- trata influenced by different solvents. J Food Sci Technol. 55(7): 2523–2532. http://jad.tums.ac.ir/