European Review for Medical and Pharmacological Sciences 2011; 15: 1010-1019 Ovicidal and repellent properties of Coccinia indica Wight and Arn. (Family:) against three important vector mosquitoes

M. GOVINDARAJAN

Division of Vector Biology and Phytochemistry, Department of Zoology, Annamalai University, Annamalainagar (India)

Abstract. – Objective: To determine the malaria, filariasis, yellow fever, dengue and larvicidal and repellent activities of Coccinia (C.) in- Japanese encephalitis, contribute significantly to dica Wight and Arnold (Family:Cucurbitaceae) ex- poverty and social debility in tropical countries1. tract against Culex (Cx.) quinquefasciatus, Aedes (Ae.) aegypti and Anopheles (An.) stephensi. Anopheles (An.) stephensi are major malaria Materials and Methods: The ovicidal activity vectors in India. With an annual incidence of was determined against three mosquito species to 300-500 million clinically manifested cases and various concentrations ranging from 50-300 ppm a death toll of 1.1-2.7 million, malaria is still under the laboratory conditions. The hatch rates one of the most important communicable dis- were assessed 48 h post treatment. The repellent eases. Currently, about 40% of the world’s popu- efficacy was determined against three mosquito lation lives in areas where malaria is endemic2. species at three concentrations viz., 1.0, 2.5 and 5.0 mg/cm2 under the laboratory conditions. Aedes (Ae.) aegypti is generally known as a vec- Results: Among five solvent extracts tested, tor for an arbovirus responsible for dengue fever, the methanol extract have most promising ovici- which is endemic to Southeast Asia, the Pacific dal activity. The methanol extract exerted zero island area, , and the Americas. This mos- hatchability (100% mortality) at 150 ppm for Cx. quito is also the vector of yellow fever in Central quinquefasciatus, at 200 ppm for Ae. aegypti and and South America and West Africa. Dengue An. stephensi. The methanol extract of C. indica found to be more repellency than the other ex- fever has become an important public health tracts. A higher concentration of 5.0 mg/cm2 pro- problem as the number of reported cases contin- vided 100% protection up to 270 min against Cx. ues to increase, especially with more severe quinquefasciatus and 210 min against Ae. aegypti forms of the disease, dengue haemorrhagic fever and An. stephensi, respectively. The results clearly and dengue shock syndrome, or with unusual show that repellent activity was dose dependent. manifestations such as central nervous system Conclusions: From the results it can be con- involvement. About two-fifths of the world’s cluded the crude extract of C. indica was an excel- lent potential for controlling Cx. quinquefasciatus, populations are now at risk of catching dengue 3 Ae. aegypti and An. stephensi mosquitoes. according to the World Health Organization . Culex (Cx.) quinquefasciatus acts as a vector for Key Words: filariasis in India. Human filariasis is a major Coccinia indica, Ovicidal activity, Repellent activity, public health hazard and remains a challenging Culex quinquefasciatus, Aedes aegypti, Anopheles socioeconomic problem in many of the tropical stephensi. countries4. Lymphatic filariasis caused by Wuchereria(W.) bancrofti and transmitted by mosquito Cx. quinquefasciatus is found to be more endemic in the Indian subcontinent. It is Introduction reported that Cx. quinquefasciatus infects more than 100 million individuals worldwide Mosquitoes not only cause nuisance by their annually5. W. bancrofti is the most predominant bites but also transmit deadly diseases like filarial nematode, which is usually characterized

Corresponding Author: Marimuthu Govindarajan, M.Sc., M.Phil., Ph.D; 1010 e-mail: [email protected] Ovicidal and repellent properties of Coccinic indica Wight and Arn by progressive debilitating swelling at the ex- tifeedant, repellent, chemosterilant, attractant, ju- tremities, scrotum, or breast (elephantiasis) in an venile and anti-juvenile hormone, moulting and infected individual6. antimoulting hormone, nematicide, rodenticide, Chemical insecticide has been used indiscrimi- fungicide and bactericide12-14. Sivagnaname and nately during the past few decades to control the Kalyanasundaram15 studied the methanolic ex- insects. This has produced serious repercussions tracts of the of Atlanta monophylla (Ru- such as insect resistance, mammalian toxicity, taceae) that were evaluated for mosquitocidal ac- bio accumulation and environment damage7. tivity against immature stages of three mosquito Toxic chemicals are responsible for contamina- species, Cx. quinquefasciatus, An. stephensi and tion of food chain and pollution of the environ- Ae. aegypti in the laboratory. Vasudevan et al16 ment. In larval mosquito control application of noted that ovicidal activity of castor oil extracted insecticide in ponds, well and other water bodies from castor seeds against mosquitoes An. may cause health hazards to human and lar- stephensi, Cx. fatigans and Ae. aegypti. Prakash17 vivorus fishes. Constant application of stated that ovicidal action of certain chitin syn- organophosphates such as temephos and fen- thesis inhibitors diflubenzuron, penfluron and thion and insect growth regulator such as di- bay SIR 8514 against mosquitoes, Cx. quinque- flubenzuron and methoprene are generally used fasciatus, Ae. aegypti, An. stephensi and An. culi- for the control of mosquito larvae8. Mosquito re- cifacies. Ovicidal effects of the seed extract of pellent using people complained of ill health ef- Atriplex canescens was reported against Cx. fect and some time required medical treatment9. quinquefasciatus18. Su and Mulla19 reported that These problems have highlighted the need for the ovicidal activity of neem products the development of new strategies for selective Azadirachtin against mosquitoes Cx. tarsalis and mosquito control. Phytochemicals are advanta- Cx. quinquefasciatus. Rajkumar and Jebanesan20 geous due to their eco-safety, target-specificity, studied that ovicidal activity of Solanum triloba- not development of resistance, reduced number tum extract against Cx. quinquefasciatus and of applications, higher acceptability and suitabil- Cx. tritaeniorhynchus. ity for rural areas. Botanicals can be used as al- Yit et al21 reported benzene and methanol ex- ternative to synthetic insecticides or along with tracts of Artemisia vulgaris has been repellent other insecticides under integrated vector control activity against Ae. aegypti. The aerial parts of programmes. The product of phytochemi- the plant Kleina pendula was used in Somalia cal, which is used as insecticides for killing lar- for the repellent and insecticidal properties; the vae or adult mosquitoes or as repellents for pro- Zanthoxylum armatus, Z.alatum, Azadirachta tection against mosquito bites. Phytochemicals indica and Curcuma aromatica were possess re- obtained from the whole plant or specific part of pellent properties against mosquitoes22;the re- the plant by the extraction with different types of pellency effect of three viz., fever tea solvent such as aqueous, methanol, chloroform, (Lippia javanica), rose geranium (Pelargonium benzene, ethyl acetate and acetone, etc., depend- reniforme) and lemongrass (Cymbopogan exca- ing on the polarity of the phytochemical. Some vatus) against laboratory reared An. arabiensis phytochemicals act as toxicant (insecticide) both mosquitoes23. Coccinia (C.) indica Wight and against adult as well as larval stages of mosqui- Arnold (Cucurbitaceae) commonly known as toes, While other interfere with growth and “Ivy gourd” and “Kovai” in Tamil is a perennial growth inhibitor or with reproduction or produce tendril climber, available in wild and cultivated an olfactory stimuli thus acting as repellent or form. It is the native of Central Africa, India attractant10. and Asia and distributed naturally in China, More than 1005 plant species are found to Tropical Asia, India, Australia and Africa. possess insecticidal properties, 384 contain an- Every part of this plant is valuable in medicine tifeedants, 297 contain repellents, and 27 contain for ring worm, psoriasis, small pox, scabies24 attractants and possess growth inhibitors11. All and other itchy skin eruptions and ulcers25. C. these indicate that the plant kingdom is a vast indica has antidiabetic26, hypoglycemic27, anti- storehouse of potentially useful chemicals for inflammatory28, analgesic29, hepatoprotective29, pest control. It is believed that insect resistance antioxidant30, antilitihic31, and antimutagenic32 likely to occur less because many botanicals con- activities. The aim of the present study was to tain multiactive principles. The pest control prin- determine the effect of ovicidal and repellent ciples include properties of insecticide, an- activities of the plant leaf extract of C. indica

1011 M. Govindarajan against the malaria (An. stephensi), dengue (Ae. Laboratory, Annamalai University, India. The dif- aegypti) and filariasis (Cx. quinquefasciatus) ferent leaf extract diluted in the appropriate sol- vector mosquitoes. vent to achieve various concentrations ranging from 50 to 300 ppm. Eggs of these mosquito species (100 nos.) were exposed to each concen- trations of leaf extract. After treatment the eggs Materials and Methods from each concentration were individually trans- ferred to distilled water cups for hatching assess- Collection of Plants ment after counting the eggs under microscope. Fully developed leaves of the C. indica were Each experiment was replicated six times along collected from different regions of Cuddalore with appropriate control. The hatch rates were as- District, Tamilnadu, India. It was authenticated sessed 48 h post treatment by following formula. by a plant taxonomist from the Department of Botany, Annamalai University. A voucher speci- Number of hatched larvae men is deposited at the Herbarium of Plant Phy- % of egg mortality = ––––––––––––––––––––––––––––––––––––––––– × 1 0 0 tochemistry Division, Department of Zoology, Total no. of eggs/egg raft Annamalai University, India. Repellent Activity Extraction The repellent study was following the method The leaves were washed with tap water, shade of WHO33. Three-day-old blood-starved female dried and finely ground. The finely ground plant Cx. quinquefasciatus, Ae. aegypti and An. material (3.0 kg/solvent) was loaded in Soxhlet stephensi mosquitoes (100) were kept in a net apparatus and was extracted with five different cage (45 cm × 30 cm × 45 cm). The volunteer solvents namely benzene, hexane, ethyl acetate, had no contact with lotions, perfumes or per- methanol and chloroform individually. The sol- fumed soaps on the day of the assay. The arms of vent from the extract was removed using a rotary volunteer, only 25 cm2 dorsal side of the skin on vacuum evaporator to collect the crude extract. each arms was exposed and the remaining area The crude residue of this plant varies with the covered by rubber gloves. The crude extract was solvents used. The C. indica with five different applied at 1.0, 2.5 and 5.0 mg/cm2 separately in solvents yielded 102.5, 97.3, 115.7, 113.5 and the exposed area of the fore arm. Only ethanol 129.3 gm of crude residue respectively. Standard served as control. The time of the test dependent stock solutions were prepared at 1% by dissolv- on whether the target mosquitoes day-or night ing the residues in acetone. From this stock solu- biters. Ae. aegypti was tested during the day time tion, different concentrations were prepared and from 07.00 to 17.00h, while Cx. quinquefasciatus these solutions were used for ovicidal and repel- and An. stephensi were tested during the night lent activity. from 19.00 to 05.00h. The control and treated arm were introduced simultaneously in to the Test Organisms mosquito cage, and gently tapping the sides on The mosquitoes, Cx. quinquefasciatus, the experimental cages, the mosquitoes were ac- An.stephensi and Ae. aegypti were reared in the tivated. Each test concentration was repeated six vector control laboratory, Department of Zoolo- times. The volunteer conducted their test of each gy, Annamalai University, India. The larvae were concentration by inserting the treated and control fed on dog biscuits and yeast powder in the 3:1 arm in to the same cage for one full minute for ratio. Adults were provided with 10% sucrose so- every five minutes. The mosquitoes that landed lution and one week old chick for blood meal. on the hand were recorded and then shaken off Mosquitoes were held at 28 ± 20, 70-85% relative before imbibing any blood; making out a 5 min- humidity (RH), with a photo period of 14 h light, utes protection. The percentage of repellency was 10 h dark. calculated by the following formula. × Ovicidal Activity % Repellency= [(Ta – Tb)/Ta] 100 Ovicidal activity was assessed by the slightly 19 modified method of Su and Mulla . The egg Where Ta is the number of mosquitoes in the raft/eggs of Cx.quinquefasciatus, An. stephensi control group and Tb is the number of mosquitoes and Ae.aegypti were collected from Vector Control in the treated group.

1012 Ovicidal and repellent properties of Coccinic indica Wight and Arn

Statistical Analysis Ae.aegypti and An.stephensi. The results from the The average larval mortality data were sub- skin repellent activity of C. indica leaf extract jected to Probit Analysis for calculating LC50, against Cx. quinquefasciatus, Ae. aegypti and An. LC90 and other statistics at 95% confidence limits stephensi are given in Table II-IV. The repellent ac- of upper confidence limit and lower confidence tivity was very high at the initial stage of exposure. limit, and chi-square values were calculated us- Increase in the exposure period showed reduction ing the SPSS12.0 (Statistical Package of Social in repellent activity and it depends upon the con- Sciences) software (SPSS Inc., Chicago, IL, centration of the extract and density of mosquito. USA). Results with p<0.05 were considered to The methanol extract of C. indica found to be be statistically significant. more effective than the other extracts. A higher concentration of 5.0 mg/cm2 provided 100% pro- tection up to 270 min against Cx. quinquefasciatus and 210 min against Ae. aegypti and An. stephensi, Results respectively. The C. indica gave the maximum pro- tection time against Cx. quinquefasciatus than Ae. The leaf extract of C. indica have been studied aegypti and An. stephensi. The results clearly show for use as natural insecticides instead of organic that repellent activity was dose dependent. phosphorous materials or other synthetic agents. Results on the ovicidal and repellent effects of leaf extract was reported in the present study, confirm their potential for control of the mosqui- Discussion to populations. Table I shows the mean per cent hatchability of Cx. quinquefasciatus, Ae. aegypti Today, the environmental safety of an insecti- and An. stephensi. All the five solvent extracts cide is considered to be of paramount importance. tested the methanol extract have most promising An insecticide does not have to cause high mortali- ovicidal activity. The methanol extract exerted ty on target organisms in order to be acceptable34. zero hatchability (100% mortality) at 150 ppm Phytochemicals may serve as suitable alternatives for Cx. quinquefasciatus, at 200 ppm for Ae. ae- to synthetic insecticides in future as they are rela- gypti and An. stephensi. tively safe, inexpensive, and are readily available in Cx. quinquefasciatus eggs were more suscepti- many areas of the world. According to Bowers et ble to the C. indica leaf extract than those of al35 the screening of locally available medicinal

Table I. Ovicidal activity of Coccinia indica plant extracts against Culex quinquefasciatus, Anopheles stephensi and Aedes aegypti.

Percentage of egg hatchability, Concentration (ppm) Name of Mosquito the solvent Control 50 100 150 200 250 300

Culex quinquefasciatus Hexane 100 ± 0.0 47.2 ± 1.4 41.6 ± 1.2 29.4 ± 0.6 15.8 ± 1.6 NH NH Ethyl acetate 99.6 ± 1.2 42.6 ± 1.8 35.8 ± 0.8 26.6 ± 1.2 NH NH NH Benzene 100 ± 0.0 38.9 ± 1.2 32.4 ± 1.2 21.2 ± 0.8 NH NH NH Chloroform 100 ± 0.0 36.6 ± 0.8 29.2 ± 1.8 17.4 ± 1.4 NH NH NH Methanol 98.3 ± 1.6 27.3 ± 0.6 18.6 ± 1.4 NH NH NH NH Anopheles stephensi Hexane 100 ± 0.0 59.2 ± 1.2 48.6 ± 1.6 39.2 ± 1.2 28.2 ± 1.4 17.6 ± 1.8 NH Ethyl acetate 100 ± 0.0 50.6 ± 1.8 42.9 ± 0.8 36.4 ± 1.6 22.6 ± 1.8 NH NH Benzene 100 ± 0.0 44.2 ± 1.6 37.4 ± 1.2 27.6 ± 0.6 16.8 ± 1.6 NH NH Chloroform 99.3 ± 0.8 39.4 ± 0.8 32.6 ± 1.6 21.9 ± 1.2 13.4 ± 1.6 NH NH Methanol 100 ± 0.0 34.4 ± 1.4 25.9 ± 1.8 14.2 ± 1.2 NH NH NH Aedes aegypti Hexane 98.9 ± 1.8 68.2 ± 1.4 57.8 ± 1.6 45.6 ± 1.6 38.4 ± 0.8 22.6 ± 1.2 NH Ethyl acetate 100 ± 0.0 59.8 ± 1.2 48.4 ± 1.4 38.2 ± 1.2 29.8 ± 1.4 NH NH Benzene 100 ± 0.0 52.8 ± 1.6 40.9 ± 0.8 31.2 ± 1.4 23.4 ± 1.2 NH NH Chloroform 97.6 ± 1.2 48.4 ± 0.8 35.4 ± 1.2 26.6 ± 1.4 18.2 ± 1.6 NH NH Methanol 100 ± 0.0 41.8 ± 2.2 29.2 ± 1.2 19.6 ± 1.2 NH NH NH

NH: No hatch ability.

1013 M. Govindarajan . Repellency % ± SD, Time of post application (minutes) Repellency % ± SD, Time Culex quinquefasciatus Culex against Coccinia indica ) 60 90 120 150 180 210 240 270 2 2.55.02.55.0 100 ± 0.02.5 100 ± 0.05.0 100 ± 0.0 100 ± 0.0 100 ± 0.02.5 100 ± 0.05.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.02.5 100 ± 0.0 100 ± 0.05.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 86.4 ± 1.2 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 64.8 ± 2.2 100 ± 0.0 100 ± 0.0 88.4 ± 1.8 81.3 ± 2.6 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 57.3 ± 1.5 100 ± 0.0 100 ± 0.0 72.6 ± 1.6 100 ± 0.0 67.8 ± 1.2 91.4 ± 1.4 84.3 ± 1.9 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 63.7 ± 1.4 79.5 ± 1.6 100 ±0.0 72.3 ± 1.2 91.2 ± 1.2 89.7 ± 1.2 100 ± 0.0 100 ± 0.0 100 ± 0.0 68.4 ± 1.2 86.9 ± 1.8 77.1 ± 1.8 100 ± 0.0 93.8 ± 1.4 100 ± 0.0 72.6 ± 1.6 100 ± 0.0 88.4 ± 1.9 100 ± 0.0 92.2 ± 1.7 100 ± 0.0 Repellency of different solvent extracts of extracts solvent of different Repellency Name of Concentration the solvent (mg/cm HexaneEthyl acetateBenzene 1.0 1.0ChloroformMethanol 100 ± 0.0 1.0 100 ± 0.0 1.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 1.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 74.6 ± 1.6 100 ± 0.0 100 ± 0.0 79.8 ± 2.2 100 ± 0.0 61.4 ± 1.7 100 ± 0.0 100 ± 0.0 68.7 ± 1.8 100 ± 0.0 51.6 ± 2.2 100 ± 0.0 82.6 ± 0.8 54.2 ± 1.6 100 ± 0.0 83.4 ± 1.9 71.8 ± 1.4 100 ± 0.0 74.6 ± 2.2 59.4 ± 0.8 100 ± 0.0 67.9 ± 1.2 100 ± 0.0 80.2 ± 1.2 Table II. Table

1014 Ovicidal and repellent properties of Coccinic indica Wight and Arn Repellency % ± SD, Time of post application (minutes) Repellency % ± SD, Time Aedes aegypti. against Coccinia indica ) 60 90 120 150 180 210 240 270 2 2.55.02.55.0 100 ± 0.02.5 100 ± 0.05.0 100 ± 0.0 100 ± 0.0 100 ± 0.02.5 100 ± 0.05.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.02.5 100 ± 0.0 100 ± 0.05.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 82.2 ± 1.8 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 71.2 ± 1.9 100 ± 0.0 86.8 ± 1.5 100 ± 0.0 100 ± 0.0 86.4 ± 1.4 100 ± 0.0 100 ± 0.0 100 ± 0.0 59.4 ± 2.4 100 ± 0.0 100 ± 0.0 100 ± 0.0 77.6 ± 2.2 78.3 ± 1.9 89.6 ± 1.2 100 ± 0.0 89.6 ± 1.8 100 ± 0.0 100 ± 0.0 51.2 ± 1.6 100 ± 0.0 68.4 ± 1.4 100 ± 0.0 62.1 ± 2.4 100 ± 0.0 81.4 ± 0.8 78.8 ± 1.2 92.6 ± 1.8 91.6 ± 1.2 100 ± 0.0 100 ± 0.0 58.6 ± 1.7 73.2 ± 2.4 100 ± 0.0 69.4 ± 1.6 87.4 ± 1.9 85.4 ± 1.8 100 ± 0.0 93.6 ± 1.2 100 ± 0.0 66.8 ± 2.2 77.4 ± 1.7 74.4 ± 1.9 93.4 ± 1.3 88.2 ± 1.3 100 ± 0.0 69.8 ± 1.3 85.6 ± 1.5 81.4 ± 1.6 91.4 ± 1.8 78.8 ± 1.6 86.2 ± 1.8 Repellency of different solvent extracts of extracts solvent of different Repellency Name of Concentration the solvent (mg/cm HexaneEthyl acetateBenzene 1.0 1.0ChloroformMethanol 100 ± 0.0 1.0 100 ± 0.0 1.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 1.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 74.4 ± 1.2 100 ± 0.0 100 ± 0.0 79.3 ± 0.8 100 ± 0.0 67.8 ± 1.6 100 ± 0.0 100 ± 0.0 71.2 ± 1.6 100 ± 0.0 58.6 ± 2.2 100 ± 0.0 82.4 ± 1.7 63.3 ± 1.8 100 ± 0.0 47.2 ± 1.8 86.2 ± 1.3 76.2 ± 1.4 49.5 ± 1.2 100 ± 0.0 79.6 ± 1.5 69.6 ± 2.2 100 ± 0.0 71.6 ± 1.5 57.4 ± 1.4 87.6 ± 1.5 64.2 ± 1.7 79.4 ± 1.2 71.6 ± 2.2 Table III. Table

1015 M. Govindarajan . Repellency % ± SD, Time of post application (minutes) Repellency % ± SD, Time Anopheles stephensi against Coccinia indica ) 60 90 120 150 180 210 240 270 2 2.55.02.55.0 100 ± 0.02.5 100 ± 0.05.0 100 ± 0.0 100 ± 0.0 100 ± 0.02.5 100 ± 0.05.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.02.5 100 ± 0.0 100 ± 0.05.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 74.4 ± 1.7 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 57.4 ± 1.8 100 ± 0.0 78.4 ± 1.2 100 ± 0.0 100 ± 0.0 72.8 ± 1.9 100 ± 0.0 100 ± 0.0 100 ± 0.0 45.6 ± 1.9 100 ± 0.0 100 ± 0.0 100 ± 0.0 68.2 ± 2.4 61.4 ± 1.5 81.4 ± 1.2 100 ± 0.0 77.8 ± 1.6 100 ± 0.0 100 ± 0.0 26.4 ± 1.6 100 ± 0.0 56.6 ± 2.8 100 ± 0.0 42.6 ± 1.2 100 ± 0.0 76.2 ± 1.4 69.4 ± 1.4 86.4 ± 1.4 81.5 ± 1.8 100 ± 0.0 100 ± 0.0 37.2 ± 1.3 64.4 ± 1.7 100 ± 0.0 57.4 ± 1.8 79.6 ± 1.9 75.2 ± 1.4 100 ± 0.0 85.4 ± 1.5 100 ± 0.0 48.6 ± 1.2 71.2 ± 1.2 66.4 ± 1.8 86.6 ± 1.8 82.6 ± 1.6 100 ± 0.0 59.4 ± 2.2 79.4 ± 1.7 71.2 ± 1.7 88.2 ± 1.4 68.2 ± 1.7 79.6 ± 1.2 Repellency of different solvent extracts of extracts solvent of different Repellency Name of Concentration the solvent (mg/cm HexaneEthyl acetateBenzene 1.0 1.0ChloroformMethanol 100 ± 0.0 1.0 100 ± 0.0 1.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 1.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 100 ± 0.0 69.6 ± 1.3 100 ± 0.0 100 ± 0.0 71.4 ± 1.4 100 ± 0.0 51.6 ± 1.5 100 ± 0.0 100 ± 0.0 59.5 ± 2.2 100 ± 0.0 39.8 ± 2.2 100 ± 0.0 76.6 ± 0.8 47.6 ± 2.2 100 ± 0.0 21.3 ± 1.8 79.3 ± 1.9 67.4 ± 1.6 28.3 ± 1.7 100 ± 0.0 71.4 ± 1.2 58.6 ± 1.3 100 ± 0.0 66.6 ± 1.9 43.8 ± 1.6 77.4 ± 1.3 52.2 ± 1.2 68.4 ± 1.2 59.4 ± 1.6 Table IV. Table

1016 Ovicidal and repellent properties of Coccinic indica Wight and Arn plants for mosquito control would generate local 1957. DEET-based products include a plasticizer, employment, reduce dependence on expensive im- capable of dissolving watch crystals, the frames ported products and stimulate local efforts to en- of glasses, and certain synthetic fabrics. Continu- hance public health. The mosquitocidal activities ous application of DEET causes infolding of the of the crude leaf extract results were also compara- epidermis with fewer hairs and a thickened der- ble with earlier reports. Su and Mulla19 reported mis with more vascularity40. Venkatachalam and that the egg rafts aged for 0, 4, 8, 12 and 24 hr Jebanesan41 have also reported that the repellent were exposed to 10 ppm neem suspensions for 36 activity of methanol extract of Ferronia elephan- hrs and the ovicidal activity was only attained in tum leaves against Ae. aegypti activity at 1.0 and the egg rafts deposited directly (0 hr old) in neem 2.5 mg/cm2 concentrations gave 100% protection suspensions, not in those with ages of 4-24 hr. In up to 2.14±0.16 h and 4.00±0.24 h, respectively, this study, the exposure period also played a crucial and the total percentage protection was 45.8% at role in causing toxicity. The ovicidal activity of 1.0 mg/cm2 and 59.0% at 2.5 mg/cm2 for 10 h. Moschosma polystachyum leaf extract against the The essential oil of Zingiber officinalis showed egg rafts of Cx. quinquefasciatus showed 100% repellent activity at 4.0 mg/cm2, which provided mortality at 0-3 h and 3-6 h with concentrations of 100% protection up to 120 min against Cx. quin- 125, 150, 175 and 200 mg/l12. quefasciatus42. In my study the leaf extract did This ovicidal and repellent activity is compa- not cause any such of discomfort or skin irrita- rable to previously screened plants in our labora- tion to the volunteers. The finding of the present tory using different species of mosquitoes. The investigation revealed that the leaf extract of C. leaf extract of Cassia fistula with different sol- indica possess ovicidal and skin repellent activity vens viz, methanol, benzene and acetone were against Cx. quinquefasciatus, Ae. aegypti and An. studied for the larvicidal and repellent activity stephensi. against Ae. aegypti. The 24 h LC50 values of the The biological activity of the plant extract extract against Ae. aegypti were observed at might be due to a variety of compounds in this 10.69, 18.27 and 23.95 mg/l respectively. The plant including phenolics, terpenoids, and alka- crude extract of C. fistula shows significant re- loids. These compounds may jointly or indepen- pellency against Aedes aegypti36; the crude leaf dently contribute to cause ovicidal and skin re- extracts of Pemphis acidula were evaluated for pellent activity against Cx. quinquefasciatus, Ae. larvicidal, ovicidal and repellent activities against aegypti and An. stephensi. These findings suggest

Cx. quinquefasciatus and Ae. aegypti. The LC50 that the leaf methanol extract of C. indica have values of methanol, benzene, acetone were the potential for use to control mosquitoes. Fur- 10.81, 41.07, 53.22 ppm and 22.10, 43.99, 57.66 ther studies are in progress to evaluate the effect ppm, respectively. Skin repellent test at 1.0, 2.5 of purified extract on mosquitocidal activity. The and 5.0 mg/cm2 concentration of P.acidula gave purified plant metabolite of the leaf methanol ex- 100% protection up to 2.30, 4.00 and 6.45 hrs tracts of C. indica may be used as environment and 2.45, 4.30 and 7.0 hrs respectively37. The leaf friendly and sustainable insecticides to combat extract of Acalypha indica with different solvents mosquitoes. In our view, biopesticides from plant viz, benzene, chloroform, ethyl acetate and origin may contribute to an effective vector con- methanol were tested for larvicidal and ovicidal trol tools. These new agents should preferentially activity against An. stephensi. The LC50 values to be applied in integrated control strategies to are 19.25, 27.76, 23.26 and 15.03 ppm, respec- gain maximum impact on adult mosquito popula- tively. The percent hatchability was inversely tions. proportional to the concentration of extract and directly proportional to the eggs38; the larvicidal –––––––––––––––––––– and repellent activities of Sida acuta extract Acknowledgements against Culex quinquefasciatus, Aedes aegypti The author is thankful to the Department of Science and Anopheles stephensi, extract had strong re- and Technology (DST) (SERC-Fast Track Young Sci- pellent action against three species of mosquitoes entist Project), New Delhi, India for providing finan- as it provided 100% protection against An. cial assistance for the present investigation. The Au- stephensi for 180 min followed by Ae. Aegypti thor is grateful to the Dr. (Mrs) Selvi Sabhanayakam, 39 Professor and Head, Department of Zoology, Anna- (150 min) and Cx. quinquefasciatus (120 min) . malai University for the laboratory facilities provided. The insect repellent that is widely available is I acknowledge the staff members of the VCRC DEET, which has been used worldwide since (ICMR), Pondicherry for their cooperation.

1017 M. Govindarajan

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