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Differential larvicidal efficacy of four species of Vitex against Culex quinquefasciatus larvae

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Differential larvicidal efficacy of four species of Vitex against Culex quinquefasciatus larvae
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  SHORT COMMUNICATION Differential larvicidal efficacy of four speciesof   Vitex   against  Culex quinquefasciatus  larvae Krishnan Kannathasan  &  Annadurai Senthilkumar  & Manivachagam Chandrasekaran  & Venugopalan Venkatesalu Received: 23 July 2007 /Accepted: 1 August 2007 / Published online: 15 August 2007 # Springer-Verlag 2007 Abstract  The early fourth instar larvae of   Culex quinque- fasciatus , reared in the laboratory were used for larvicidalassay with leaf extracts of   Vitex negundo, Vitex trifolia, Vitex peduncularis  and  Vitex altissima . The methanol extracts of the four species possessed varying levels of larvicidal nature.The highest larvicidal activity was found with the extract of  V. trifolia  (LC 50 =41.41 ppm) followed by  V. peduncularis (LC 50 =76.28 ppm),  V. altissima  (LC 50 =128.04 ppm) and  V.negundo  (LC 50 =212.57 ppm). Introduction The vector-borne diseases caused by mosquitoes are one of the major health problems in many countries. Dengue,malaria, yellow fever, filariasis and chickungunya are someof the deadly diseases spread by mosquitoes. Lymphaticfilariasis is caused by  Wucherria bancrofti , and this pathogenis transmitted by  Culex quinquefaciatus . The control of themosquitoes has become complicated because of their resistance to synthetic insecticides. In recent years, inves-tigations on botanicals are highly targeted for the identifica-tion of plant chemicals for the control of mosquitoes (Ansariet al. 2005; Singh et al. 2006; de Omena et al. 2007). The genus  Vitex  belongs to the family Verbenaceae, andits species are shrubs or trees, present in the tropical andtemperate regions of the world. India, 1 of the 11 mega- biodiversity countries, has 13 species of   Vitex  (Rajendranand Daniel 2002). The  Vitex  species are used in the Indiansystem of medicine for the treatment of a range of diseases. Vitex negundo  and  Vitex trifolia  are prescribed to be major herbal plants among the  Vitex  species. V. negundo  is a shrub or small tree (4  –  8 m) locally knownas  “  Nochi ”  by the people of Tamilnadu. The leaves of this plant are aromatic and effective in dispelling inflammatoryswelling of joints in acute inflammation and also in splenicenlargement. In Ayurvedic medicine, a reputed drug namely, “  Nirgundi ”  is prepared from  V. negundo , and the drug isreportedly astringent, bitter and cold. It improves receptiveand retentive power of mind and complexion of growth of hair (Sivarajan and Balachandran 2002). The decoction of the leaves is used in bath during puerperal state of women inIndia. The juice made from fresh leaves is poured into thenostrils in stupor and coma (Kirtikar and Basu 1991). Invillages of Tamilnadu, people use the dried aerial parts of  V. negundo  during the storage of paddy after harvesting.They keep the plant over paddy or rice bags to protect frominsects. V. trifolia  is a shrub 3  –  6 m high. Because this plant can be commonly seen on the banks of water bodies likechannels, rivers and ponds, it is locally called as  “  Neer  Nochi ”  (In Tamil language  “  Neer  ”  means water). Theleaves of   V. trifolia  improve memory, useful in pain,remove bad taste in mouth and cure fever (Kirtikar andBasu 1991). The leaf extract is anti-cancerous. The flowersare useful in fevers and fruits are good for amenorrhoea.The leaves are used to treat hair loss (Varier  2003). Theflowers of   V. trifolia  mixed with honey are useful in feversaccompanied by vomiting and severe thirst (Bhattacharjeeand De 2005). The aerial parts of this plant are useful in thetreatment of diabetes (Pullaiah and Naidu 2003).The  Vitex peduncularis  is a tree about 18 m tall. The plant is known as  “ Mayiladi ”  or   “ Malainochi ”  by the local people. The bark of this plant is used to cure pains in the Parasitol Res (2007) 101:1721  –  1723DOI 10.1007/s00436-007-0714-5K. Kannathasan : A. Senthilkumar  :  M. Chandrasekaran : V. Venkatesalu ( * )Department of Botany, Annamalai University,Annamalai Nagar 608 002 Tamil Nadu, Indiae-mail: venkatesalu@yahoo.com  chest. Leaves, root bark or young stem bark are useful inmalarial fever (Kirtikar and Basu 1991). In traditionalmedicine of Mizoram, India, 10  –  15 ml of leaf, root or bark decoction is taken orally in the treatment of malaria(Sharma et al. 2001). The  Vitex altissima  is called as “ Mayla ”  in Tamilnadu, and this plant is reported to possessanti-inflammatory activity (Sridhar et al. 2005).In this study, the larvicidal efficacy of four species of  Vitex , namely,  V. negundo ,  V. trifolia, V. peduncularis  and V. altissima  was examined against the fourth instar larvae of  Culex quinquefasciatus . Materials and methods Plant materialTheleavesof  V.negundo Linn., V.trifolia  Linn., V.peduncularis Wall. ex Schauer and  V. altissima  Linn. f. were collected fromdifferent regions of Tamilnadu, India. The voucher specimensare deposited at the herbarium, Department of Botany,Annamalai University. The place of collection, altitude andits geographical location are given in Table 1.The leaf samples were shade dried followed by ovendrying (60°C). The dried leaves were milled in an electrical blender, and 30 g powder was extracted with methanol(100 ml×3) by Soxhlet apparatus for 72 h. The crudeextracts were dried under vacuum in a rotary evaporator at 40°C. The extract was stored below 4°C until further assay.Larvicidal assayThe egg rafts of   C. quinquefasciatus  were collected fromdrainage of local residential area of Annamalainagar (11°23 ′ 17N, 79°42 ′ 57E) and reared in laboratory (29±3°C,75 to 85% RH). The larvae were fed with Brewer  ’ s yeast:dog biscuit (1:3). The larvae at early fourth instar stage wereused for larvicidal assay.The larvicidal activity was analysed as per the standard procedures recommended by World Health Organisation(1981). The plant extracts were dissolved in 2 ml of dimethyl sulfoxide and prepared into different concentra-tions like 50, 100, 200, 300, 400 and 500 ppm with distilledwater. Twenty larvae were taken in a class beaker (250 ml)containing 199 ml of tap water and 1 ml of respectiveconcentration of plant extract. Five replicates were main-tained for each concentration, and the dead larvae werecounted after 24 h. The lethal concentration, LC 50  at whichconcentration (ppm) 50% of larvae showed mortality, 95%confidence limit of upper and lower confidence levels werecalculated by probit analysis (SPSS, version 11.5). Results and discussion The larvicidal efficacy of four species of   Vitex  is presented inTable 2. Among the four extracts, the extract of   V. trifolia  wasfound highly active against the larvae of   C. quinquefasciatus (LC 50 =41.41 ppm). The leaf extract of   V. negundo  showedthe lowest LC 50  (212.57 ppm) value.The essential oil of   V. negundo  leaves was found to havelarvicidal activity against   Aedes aegypti  mosquito larvae(Hebbalkar et al. 1992). However, in this investigation, theleaf extract of   V. trifolia  possessed higher activity than V. negundo .Generally, the  Vitex  species are proved for their richmedicinal uses.  V. trifolia, V. polygama  and  V. mollis  showedinsecticidal ability against fall army worm  Spodoptera frugiperda  (Hernandez et al. 1999; Gallo et al. 2006; Rodríguez-López et al. 2007). The alcoholic-based seedextract of chaste tree ( V. agnus castus ) showed mosquitorepellent efficacy against three genera of mosquitoes viz.,  Aedes, Anopheles  and  Culex  (Mehlhorn et al. 2005). The V. trifolia , which showed more potent activity is called  “ threeleaved chaste tree ”  in India (Varier  2003). Table 1  Geographical details of place of sample collection Name of species Place of collection Geographical location Altitude (m) Vitex negundo  Kollidam −  Nagapattinam district 11°18 ′ 47N 079°42 ′ 04E 13 Vitex trifolia  Sivapuri − Cuddalore district 11°24 ′ 24N 079°42 ′ 59E 18 Vitex peduncularis  Jamanamarathur, Javadhu Hills  –   Tiruvannamalai district 12°36 ′ 10N 078°53 ′ 07E 705 Vitex altissima  Chitheri, Chitheri Hills  –  Dharmapuri district 11°53 ′ 28N 078°30 ′ 26E 959 Table 2  Larvicidal efficacy of four species of   Vitex  against   C.quinquefasciatus  Name of species LC 50  95% Confidence limit UCL LCL Vitex negundo  212.57 251.57 171.70 Vitex trifolia  41.41 63.50 9.25 Vitex peduncularis  76.28 100.37 45.21 Vitex altissima  128.04 187.67 37.17 UCL  upper confidence level,  LCL  lower confidence level1722 Parasitol Res (2007) 101:1721  –  1723  In the present study,  V. peduncularis  exhibited amoderate larvidical activity (LC 50 =76.28 ppm). The polar fractions (butanol) of this plant contained an iridoid,namely, pedunculariside with anti-inflammatory activity(Suksamrarn et al. 2002).This study showed a clear comparison in the larvicidalnatureoffourspeciesof  Vitex , namely,  V. negundo, V. trifolia,V. peduncularis  and  V. altissima . Further investigation on thefield applications of these extracts and also the isolation of active principle compound involved in the killing of larvaewill be useful for the control of   C. quinquefasciatus  bornefilariasis. Acknowledgement  The authors grateful to Dr. R. Panneerselvam,Professor and Head, Department of Botany for having provided thelaboratory facility for this study. References Ansari MA, Mittal PK, Razdan RK, Sreehari U (2005) Larvicidal andmosquito repellent activities of pine (  Pinus longifolia,  Family:Pinaceae) oil. J Vect Borne Dis 42:95  –  99Bhattacharjee SK, De LC (2005) Medicinal herbs and flowers.Avishkar, Jaipur, India, p 306de Omena MC, Navarro DMAF, de Paula JE, Luna JS, de Lima FMR,Sant  ’ Ana AEG (2007) Larvicidal activities against   Aedes aegypti of some Brazilian medicinal plants. Biores Technol 98:2549  –  2556Gallo MBC, Rocha WC, Da Cunha US, Diogo FA, DA Silva FC, VieiraPC, Vendramim JD, Fernandes JB, Da Silva MFGF, Batista-PereiraLG(2006)Bioactivityofextractsandisolatedcompoundsfrom Vitex polygama  (Verbenaceae) and  Siphoneugena densiflora  (Myrtaceae)against   Spodoptera frugiperda  (Lepidoptera: Noctuidae). Pest Manag Sci 62:1072  –  1081Hebbalkar DS, Hebbalkar GD, Sharma RN, Joshi VS, Bhat VS (1992)Mosquito repellent activity of oils from  Vitex negundo  Linn.leaves. Indian J Med Res 95:200  –  203Hernandez MM, Heraso C, Villareal ML, Vargas-Arispuro I, Aranda E(1999) Biological activities of crude plant extracts from  Vitextrifolia  L. (Verbenaceae). J Ethnopharmacol 67:37  –  44Kirtikar KR, Basu BD (1991) Indian medicinal plants, Lalit MohanBasu, Allahabad, India, pp 1935  –  1944Mehlhorn H, Schmahl G, Schmidt J (2005) Extract of the seeds of the plant   Vitex agnus castus  proven to be highly efficacious as a re- pellent against ticks, fleas, mosquitoes and biting flies. ParasitolRes 95:363  –  365Pullaiah T, Naidu KC (2003) Antidiabetic plants in India and herbal bases antibiotic research. Recency, New Delhi, pp 314  –  315Rajendran A, Daniel P (2002) The Indian Verbenaceae-A taxonomicrevision, Bishen Singh Mahendra Pal Singh, Dehradun, pp 341  –  386Rodríguez-López V, Figueroa-Suàrez MZ, Rodríguez T, Aranda E(2007) Insecticidal activity of   Vitex mollis.  Fitoterapia 78:37  –  39Sharma HK, Chhangte L, Dolui AK (2001) Traditional medicinal plants in Mizoram, India. Fitoterapia 72:146  –  161Singh RK, Dhiman RC, Mittal PK (2006) Mosquito larvicidal propertiesof   Momordica charantia  Linn. (Family: Cucurbitaceae). J Vect Borne Dis 43:88  –  91Sivarajan VV, Balachandran I (2002) Ayurvedic drugs and their plant sources. Oxford and IHB, New Delhi, pp 329  –  332Sridhar C, Rao KV, Subbaraju GV (2005) Flavonoids, triterpenoidsand a lignan from  Vitex altissima.  Phytochemistry 66:1707  –  1712Suksamrarn A, Kumpun S, Kiritikara K, Yingyongnarongkul B,Suksamrarn S (2002) Iridioids with anti-inflammatory activityfrom  Vitex peduncularis . Planta Med 68:72  –  73Varier PS (2003) Indian medicinal plants. Orient Longman, Hyderabad,India, pp 387  –  395World Health Organisation (1981) Instruction for determining thesusceptibility or resistance of mosquito larvae to insecticides.WHO-VBC 81.807, pp 1  –  6Parasitol Res (2007) 101:1721  –  1723 1723
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