jear2012 [Journal of Entomological and Acarological Research 2016; 48:4938] [page 345] Abstract The Ocimum plant was traditionally used for mosquitoes repellent and control in India especially in Tamil Nadu. In this research, deals with the larvicidal, pupicidal and adulticidal potential of three different solvent extracts of O. canum against Aedes aegypti. The overall result highlights that the chloroform extracts of O. canum were shown sig- nificant larvicidal (15.027 mg/L) activity at 24 h of exposure. The pupi- cidal and adulticidal activity of this plant exhibits highest mortality against A. aegypti within 24 h at the dose ranges of 89.773 mg/mL, 41.912 mg/mL respectively. The chloroform extracts contain major phyto-constituents like phenol, alkaloids, protein and tannins. Thin layer chromatography profiles also provide a database for the presence of active components. GC-MS analysis of bioactive chloroform extract revealed that a total of seventeen compounds, six were considered as major and the remaining as minor compounds. The spectral studies of FT-IR denoted the functional groups of bioactive components like alkenes, ketone, hydroxyl and others. Based on the outcome of results show that Ocimum canum have found to potent ability for controlling the mosquitoes, it can be used as an ideal eco-friendly agent for arrest- ing dengue fever in future. Introduction Mosquitoes are the most important single group of insects in terms of public health importance, which transmit variety of diseases like malaria, filariasis, dengue and Japanese encephalitis. It causing millions of deaths every year (Brown, 1986). Mosquito-borne diseases contribute to a larger proportion of health problems in developing countries. Repetition of synthetic insecticides for mosquito control has disrupted natural biological control systems and led to resurgences in mosquito populations. It also resulted in the development of resist- ance, undesirable effects on non-target organisms, and fostered envi- ronmental and human health concern (Thomas et al., 2004). The recent research focusing for herbal preparations that do not produce any adverse side effects in the non-target organisms and easily biodegradable (Kant et al., 1996). In general, plant derived compounds (phytopesticides) have been recognized as an important natural resources of insecticides (Gbolade et al., 2000). Several phytochemi- cals have been reported to exhibit harmful effect against mosquito lar- vae and insecticides, reproduction of inhibitors, repellent potential, ovicidal and oviposition deterrent (Prajapati et al., 2005; Pushpanathan et al., 2006) properties. One of the method available for the control of mosquito population is over and injudicious applications of persistent synthetic insecticides, resulting undesirable effect of synthetic insecticides. Previous research has been proved the effec- tiveness of plant derived secondary compounds, such as saphonine (Chowdhury et al., 2008), steroids (Ghosh et al., 2008), isoflavonoids (Josep et al., 2004), essential oil (Cavalcanti et al., 2004), alkaloids and tannins (Khanna et al., 2007) reported as mosquitocidal agents. Dengue fever is considered as a serious public health problem in the world. In tropical countries, where the favorable environmental conditions are responsible for the proliferation of vector Aedes aegyp- ti. Among the arbovirus in India, distribution of all the dengue virus type is continuously expanding. Remarkably the reemergence of Chikungunya virus (CHIK) since 2005 is posing an additional concur- rent diseases burden in the country. Aedes aegypti (L) (Diptera: Culicidae) is a fresh water breeding mosquito it is very difficult to control during rainy season.Approximately 2500 million people, two fifths of the world’s population, are now at risk from Dengue fever (Fulmali et al., 2008; Kumar et al., 2008). The WHO currently esti- mates there may be 50 million cases of Dengue fever worldwide every year (WHO, 2011). Lamiaceae have been traditionally used in developing countries for their insecticidal and repellent property against several insect Correspondence: Devarajan Natarajan, Natural Drug Research Laboratory, Department of Biotechnology, School of Biosciences, Periyar University, Salem, Tamilnadu, India. Fax: +91.427.2345124 (office). E-mail: mdnataraj@rediffmail.com ; natarajpu@gmail.com Key words: Mosquitoes repellent activity; Ocimum canum; GC-MS analysis; Aedes aegypti. Acknowledgements: the author’s express their sincere thanks to TNSCST, Tamilnadu for the finance assistance under the Student project scheme (TNSCST/SPS/TD/2013-2014). Acknowledged to VIT University (SIF) and St. Joseph college, for provide spectral analysis of GC-MS for FT-IR and HPLC. We also acknowledge the Department of Biotechnology, Periyar University, Salem 636 011 for providing laboratory facilities for baseline work. Received for publication: 6 January 2015. Revision received: 7 July 2016. Accepted for publication: 7 July 2016. ©Copyright O. Prabhavathi et al., 2016 Licensee PAGEPress, Italy Journal of Entomological and Acarological Research 2016; 48:4938 doi:10.4081/jear.2016.4938 This article is distributed under the terms of the Creative Commons Attribution Noncommercial License (by-nc 4.0) which permits any noncom- mercial use, distribution, and reproduction in any medium, provided the orig- inal author(s) and source are credited. Journal of Entomological and Acarological Research 2012; volume 44:e Journal of Entomological and Acarological Research 2016; volume 48:4938 Mosquitocidal properties of Ocimum canum Sims (Lamiaceae) leaf extracts against dengue vector Aedes aegypti L. (Diptera: Culicidae) O. Prabhavathi, R. Yuvarajan, D. Natarajan Natural Drug Research Laboratory, Department of Biotechnology, School of Biosciences, Periyar University, Salem, Tamilnadu, India No n- co mm er cia l u se on ly [page 346] [Journal of Entomological and Acarological Research 2016; 48:4938] species (Ngamo et al., 2007). For example, Rosmarinus officinalis and Lavandulaan gustifolia showed moderate larvicidal activity (Conti et al., 2010). Prajapati et al. (2005) reported the essential oils from select- ed plants as noticeable repellent and ovicidal properties, Hyptissu ave- olens has useful insecticidal (Amusan et al., 2005; Jaenson et al., 2006) and control many stored product pests (Peerzada, 1997; Othira et al., 2009; Conti et al., 2011). Moreover, its chemical composition and bio- logical activity might be changed due to the function origin and collec- tion time of plants (Noudjou et al., 2007). Ocimum canum (Lamiaceae) is a pubescent erect much branched herb, 15-60 cm high with sub-quadrangular striate branches. Leaves are elliptic-lanceolate, glabrous and gland dotted strongly aromatic herb, widely distributed in throughout India (especially in fields of waste lands, plains and lower hills). It contain several volatile oils include methyl cinnamate, methylheptenone, methylnonylketone, d- camphor, citral, ocimin, methylchavicol, linalool, nevadensin, salvi- genin, beta-sitosterol, betulinic, ursolic, oleanolic acids, flavanoids, pectolinarigenin-7-methylether and nevadensin, respectively. Polysaccharides composed of xylose, arabinose, rhamnose and galac- turonic acids (Rastogi and Mehrotra, 1993). The main biological prop- erties of O. canum reported as antimicrobial, antioxidant, anthelmintic and anti-diabetic agents (Bhattacharjee, 2001; Chopra, 1956). The present study was focused on to perform the mosquitocidal activity and identifying the potential bioactive compounds from O. canum against Dengue vector Aedes aegypti. Materials and methods Collection of plant materials The fresh leaves of O. canum were collected (during the month of November and December 2013) from Pottaneri village, Mettur (Tk), Salem District, Tamil Nadu. The plant specimen was authenticated by Dr. D. Natarajan, Assistant Professor, Department of Biotechnology, Periyar University, Salem and also cross-checked with available books and herbarium records. The voucher specimen was deposited in the NDRL for further reference. The collected plants leaves were washed with tap water to remove unwanted solid dust particles and shade - dried at room temperature for 10 days. The dried plant material was powdered separately using commercial electrical blender. Preparation of extracts The processed plant materials (500 g) were sequentially extracted by hot extraction method in a soxhlet apparatus using three organic solvents (acetone, chloroform and hexane) for 48 to 72 h until the influx solvent changed into colorless. The plant extracts were filtered through Whatman filter paper No. 1. Extracts were concentrated under reduced pressure at 40°C using rotary vacuum evaporator. The dried crude extracts were weighed for calculating their extractive value and stored in an air tight container at 4°C for further bioassays. Mosquito source Aedes aegypti (larva, pupa and adult) mosquitos were collected from NCDC, Connoor, Tamil Nadu, India. It was maintained at Natural Drug Research Laboratory, Department of Biotechnology, Periyar University, Salem. The larvae were kept in plastic trays containing tap water, and maintained at 27±2°C with 75-85% relative humidity under 14:10 h light and dark. Larvae were fed with diet of yeast, dog biscuits and sugar solution for adult mosquitos. Bioassay tests Larvicidal bioassay The larvicidal activities of selected plant crude extracts were assessed as per WHO protocol (1981). Briefly, in a container 25 fourth instar larvae were kept in 249 mL of distilled water with 1mL of differ- ent concentrations (100, 200, 300, 400 and 500 mg/L) of plant extracts. The chamber containing the control larvae received 1 mL of DMSO served as negative control. After 24 h exposure, the dead larvae were counted and corrected using Abbott’s formula and the percentage mor- tality was recorded from the average of three replicates. The average mortality percentages of three replicates were used to carry out lethal concentrations (LC50 and LC90) by Probit analysis. Pupicidal bioassay Pupicidal activities of crude extracts was evaluated as per the mod- ified method of Modify as Krishnappa et al. (2012). For the bioassay in a container, 25 pupae were kept in 249 mL of distilled water with 1 mL of extract at different concentrations (100, 200, 300, 400 and 500 mg/L) in DMSO. The container received 1 mL of DMSO served as negative control. All containers were maintained at room temperature (28±2) with naturally prevailing photoperiod (12:12 h/L:D) in the laboratory. Any pupa was considered to be dead if did not move when prodded repeatedly with a soft brush. After exposure period, the dead larvae were counted and mortality was corrected by Abbott’s (1925) formula. The percentage mortality was recorded from the average of three repli- cates (Finney, 1971). Adulticidal bioassay A total of 15 adult female mosquitoes (3-5 days old) were treated with different concentrations (100, 200, 300, 400 and 500 mg/L) of plant crude extracts impregnated with filter papers (WHOm 1981). The mos- quitoes were allowed to acclimatize in the holding tube for 1 h and then exposed to test paper for 1 h. Mortality was recorded every 10 min throughout the exposure period. Mortality of mosquitoes was deter- mined at the end of 24 h recovery period. Percentage of mortality was corrected by Abbott’s formula. LC50 and LC90 with 95% confidence limits were determined using Probit analysis (Finney, 1971). Statistical analysis The average larval (adult) mortality data were subjected to probit analysis for calculating LC50 and LC90 statistics at 95 % confidence lim- its of upper confidence limit (UCL) and lower confidence limit (LCL) values and chi square test were calculated using the SPSS 14.0. Phytochemical analysis The qualitative phytochemicals (phenol, alkaloids, quinones, glyco- sides, flavanoids, amino acids, tannins, proteins, cabhohdrate and saponin) analysis of three different extracts (acetone, chloroform, and hexane) were performed as per the methods of Behera et al. (2012). Thin layer chromatography Thin layer chromatography was performed as per the method of Bishnu et al. (2011). Silica powder was added to distilled water and mixed with magnetic stirring continuously. The slurry was poured into a clean and dried slide scattered all over the slide to make a thin film. The silica plates were activated by heating them in hot air oven at 120°C for 3 h. After 3 h, the silica plates were allowed to cool at room temperature and marked about 1cm from the bottom. The extracts were loaded at the bottom center of the slide.The beaker was saturated with suitable solvent system [methanol: chloroform (7:3)].The final solvent front was marked and the plate was dried. The developed TLC plates Article No n- co mm er cia l u se on ly were dried and visually observed for various bands. The Rf (Retension frequency) value was calculated as follows: Rf = Distance travelled by compound Distance travelled by solvent Spot visualization Few pieces of iodine crystals were kept in the iodine vapor contain- er. The plates were kept in iodine vapor and left for few hours. Brown colored bands were visualized. The bands were photographed under UV trans-illuminator (low beam and high beam UV light). High performance liquid chromatography The HPLC system was used to quantify the bioactive compounds from chloroform extracts of O. canum. The chromatographic separation was performed on a C18 column (5 mm, 250×4.6 mm i.d.) with the col- umn temperature of 35°C. Linear gradient elution was collected (with methanol and aqueous) and the procedure as follows (v/v):5 to 25% (0 to 8 min), 25 to 55% (8 to 20 min) and 55 to 100% (20 to 30 min). The UV detector is used as 280 nm (Deo et al., 2011). GC-MS analysis The GC-MS analyses of chloroform extract of plant was done by Perkin Elmer Q-700 equipment. Column temperature was programmed at 35°C-180°C for 2 min and simultaneously, increased about 4°C- 20°C/min. Helium was used as the carrier gas at 0.9 mL/min. The mass spectrum was obtained at 70 eV ionization voltage. The identification of individual compound was done using mass spectral database (the NIST (version 3.0) database). Furthermore, the Retention Time (RT) and Kovats Index (KI) values of reference compounds were compared with isolated compounds for identification Cheng et al. (2009). Fourier transform infrared spectroscopy An arid Zone FT-IR equipped with DTGS detector was used to meas- ure the spectrum developed in chloroform extracts of O. canum. FT-IR spectrum was analyzed a thin transparent oil films were made by press- ing two NaCl discs (25×5 mm) of the liquid derivatives. Absorption spec- tra were acquired at 4 cm-1 resolution and signal-averaged over 32 scans. Interferegrams were Fourier transformed using cosine apodization for optimum linear response. Spectra were baseline corrected, scaled for mass differences and normalized to the methylene peak at 2927 cm-1. Results and discussion Larvicidal activity Mortality percentage was calculated for its toxicity effect of plant crude extracts (acetone, hexane and chloroform) against fourth instar larvae of A. aegypti. The mortality percentage of larvae was calculated 24hrs of exposure. The highest larval mortality found in choloroform extract of O. canum plant against IVth instar larvae of Ades agypti with the LC50 and LC90 values of 15.027 and 353.49 mg/L. Whereas the ace- tone extract exhibits moderate activity with LC50 and LC90 values of 58.873 and 134.47 mg/L respectively (Table 1). Recently, the researcher reported that Ocimum plant extract shows better larvicidal activity against larval and adults of mosquito species (Pratheeba et al., 2015; Murugan et al., 2016). Pupicidal activity The pupal mortality of Ades agypti (after treatment of O. canum plant extract) was observed at various concentrations (Table 1). The [Journal of Entomological and Acarological Research 2016; 48:4938] [page 347] Article T ab le 1 . M o sq u it o ci d al a ct iv it y o f O . ca n u m le af e xt ra ct s ag ai n st A . ag yp ti . O . c an u m L ar vi ci da l Pu pi ci da l A du lt ic id al E xt ra ct s C on . % L C 50 LC 90 S lo pe C on . % L C 50 LC 90 S lo pe C on . % L C 50 LC 90 S lo pe M g/ L M or ta li ty M g/ L M or ta li ty M g/ L M or ta li ty Ac et on e 1 00 4 2 5 8. 87 3 1 34 .4 7 -6 .3 23 1 00 4 4 4 1. 91 2 2 04 .2 5 -3 .0 23 1 00 4 2 1 60 .7 1 4 19 .2 7 -6 .7 89 2 00 4 8 2 00 4 8 2 00 4 3 3 00 5 2 3 00 4 8 3 00 4 8 4 00 5 8 4 00 5 2 4 00 5 3 5 00 6 6 5 00 8 8 5 00 5 9 Ch lo ro fo rm 1 00 5 6 1 5. 02 7 3 53 .4 9 -1 .1 1 00 5 5 7 2. 26 5 1 19 .3 8 -1 0. 93 1 00 7 0 2 2. 66 2 1 56 .4 3 -2 .0 70 2 00 6 8 2 00 7 2 2 00 7 6 3 00 7 8 3 00 7 6 3 00 8 8 4 00 9 1 4 00 8 0 4 00 1 00 5 00 9 7 5 00 8 0 5 00 1 00 H ex an e 1 00 1 2 1 90 .5 0 3 41 .4 7 -1 1. 58 1 00 5 3 9 2. 26 5 1 59 .2 8 - 7. 93 10 0 1 2 1 08 .3 8 8 42 .3 2 -1 .3 79 2 00 2 7 2 00 5 6 2 00 3 2 3 00 4 2 3 00 7 3 3 00 5 6 4 00 5 5 4 00 7 3 4 00 6 8 5 00 5 7 5 00 7 7 5 00 6 1 Co n. , c on ce nt ra tio n; L C 5 0, le th al c on ce nt ra tio n 50 ; L C 9 0, le th al c on ce nt ra tio n 90 . No n- co mm er cia l u se on ly [page 348] [Journal of Entomological and Acarological Research 2016; 48:4938] mortality rate was increased on the basis of concentration/dose (low to high) of the extracts. The highest pupal mortality was observed in ace- tone extract with low LC50 value 41.912 mg/L respectively. Similarly, the above result was supported by various scientists like Selvakumar et al. (2015) reported that the better pupicidal activity of different solvents extracts of Annona reticulata and Gokulakrishnan et al., (2013) were identified essential oils from Pogostemon cablin and evaluated the pupicidal activity against various mosquitoes including A. aegypti. Adulticidal activity The result of adulticidal activity of chloroform leaves extract of O. canum show maximum adulticidal property with very low LC50 values (22.662 mg/L) compared with other solvents like acetone and hexane (Table 1). The results highlights the leaf and flower of O. sanctum were tested against fourth instar larvae of Aedes aegypti and they determined the LC50 values at various concentrations of extract like 425.94, 150.40, 350.78, 575.26 and 175.67 mg/L (Mohamed et al., 2008). Similarly, Govindarajan et al. (2014) reported the larvicidal, and adulticidal potential of the chloroform extract from the Erythrina indica tested against A. aegypti. The highest larval mortality and adulticidal activity were noticed in methanol extract of E. indica. Hexane leaves extracts of Citrus sinesis against the early fourth instars and female adult of Aedes aegypti. The hexane extract from C. sinensis leaves are proved to be reasonably larvicidal but remarkably irritant against dengue vector (LC50 and LC90 values of 446.84 and 1370.96 mg/L respectively) was noticed after 24 h exposure (Radhika et al., 2011). Methanolic leaf extract of S. campanulata have the potential to be used as an ideal eco- friendly approach the control of mosquitoes especially A. aegypti (Karthika Devi et al., 2013). Ethanolic and petroleum ether extracts from various parts of R. nasutus, D.elliptica, T. reidioides, H. aromatica, S. tuberose and A. calamusi were tested for their larvicidal activity potential against A. aegypti mosquitoes (Naruman Komalamisra et al., 2005). The larval toxicity and smoke repellent potential of Albizzia amara and O. basilicum at different concentrations against Aedes aegypti, resultedthat the A. amara was more effective against A. aegypti than O. basilicum (Murugan et al., 2007). The adulticidal and repellent activities of crude hexane, chloroform, benzene, acetone and methanol extracts of the leaf of Cassia tora leaves against A. aegypti. Among them, methanol extract was showed significant activity (Duraisamy Amerasan et al., 2012). Kamaraj et al. (2008) reported high larval mor- tality in methanol extracts of Cryptocoryne auriculata and Solanum torvum against the larvae of An. subpictus (LC50 44.21, 44.69, 53.16, 41.07, 35.32, 28.90 and 44.40 ppm; LC90 187.31, 188.29, 233.18, 142.66, 151.60, 121.05, 192.11 ppm, respectively) and Cx. tritaeniorhynchus (LC50 69.83, 51.29, 81.24, 71.79, 44.42, 84.47 and 65.35 ppm; LC50 335.26, 245.63, 300.45, 361.83, 185.09, 351.41 and 302.42 ppm, respectively). Bioefficacy of plant extracts differ from species to species of plants. The changes in adulticidal activity of these extracts is probably due to variation in the types and levels of active ingredients that depend not only on the genetic characteristics of the plant species but also the con- ditions under which they were grown and harvested (Tawatsin et al., 2006). Phytochemical analysis Phytochemical analysis of O. canum reveals that the acetone extracts indicate the presence of phenol, saponins, tannins and pro- teins (Table 2). Chloroform extract show the presence of phenol, flavonoids, saponins, protein and carbohydrates and the hexane extracts showed the presence of flavonoids, saponins, glycosedes and proteins. Saponins and proteins are presented in all the tested extracts. Similarly, the results were reported that the presence of volatile oils, flavonoids, carbohydrates, phytosterols, tannins and fixed oils from the leaves extracts of O. americanum (Sarma et al., 2011). The results revealed that the presence of phytochemicals viz., alkaloids, saponins, tannins, steroids, phlobatannin, terpenoids, flavinoids and cardiac gly- cosides in the Ocimumspeceis (Muhammad Neem Abbas et al., 2013). The phytochemical in the peels of R. sativuscontain most important phyto-constituents like tannins, saponins, flavonoids, amino acids, ter- penoids, cardiac glycosides and chalcones (Safia Janjua et al., 2014) and also the results were suggested in O. sanctum (Himal Paudel Chetri et al., 2008). Thin layer chromatography profile for chloroform extracts of O. canum The results of thin layer chromatography profile show different band formations (Figure 1) based on the solvent systems (methanol and chloroform) percentage of 5%, 10%, 20% and 25%, which shows dif- ferent Rf value. The Rf values are 0.038, 0.192, 0.615, 0.803 and 1cm. Based on the Rf value suggested and assumed some bioactive compo- nents using confirmation test. The qualitative and quantitative analy- sis of major constituents from O. sanctum includes eugenol, qurcetine and some others by TLC (Rawat et al., 2011) and ursolic acid (Kedar Kumar Rout et al.,2012). O. gratissimum, O. sanctum and O. canum leaves extracts were showed the better band formation with the Rf val- ues of O. canum (0.513, 0.40, 0.58, 0.38 and 0.82) referred to Lutein Pheophytin Xanthophyll Oil Chlorophyll b and �-carotene, O. sanctum (0.445, 0.59, 0.74, 0.934) and O. gratissimum (0.431, 0.573, 0.78) (Quereshi et al., 2011). Eugenol and Methyl eugenol from the petrole- um ether extracts of O. sanctum (Nasare, 2013). High performance liquid chromatography analysis of O. canum The HPLC analysis of O. canum leaf crude chloroform extracts results show major peaks (more concentration of components) at the retention times (min.) of 3.482, 5.459, 9.960 and12.688 (at wavelength of 254 nm) (Table 3 and Figure 2). The previous studies of HPLC analy- sis were reported that the presence ofeugenol (Joshi et al., 2011) and phenolic compounds and terpenes (Shanmuga Sundaram et al., 2011) identified from the leaves of O. sanctum. Similarly, the total content of bioactive compounds from the species of lamiaceae family plants, flavonoids derivatives were identified from the Thymus species (Kulevanova et al., 2001). Flavonoid content from the methanolic and aqueous extracts of O. sanctum and O. kilimandsacharicum (Deo et al., 2011), the flavonoid and total phenolic contents of methanolic extract was higher than aqueous extract of Stachys inflata (Sayyed Mehdy et al., 2011), The content of rosmarinic acid was quantified from the methanol crude extract of Perilla frutescens (Jing Liu, 2013). Article Table 2. Phytochemical analysis of crude extract of O. canum. Phytochemical testAcetone Hexane Chloroform Phenol + - + Quinons - - - Flavonoids - + + Alkaloids - - - Amino acids - - - Saponins + + + Glycosides - + - Proteins + + + Tannins + - - Carbohydrates - - + -, absence;+, presence. No n- co mm er cia l u se on ly Fourier transform infrared spectroscopy analysis of chloroform extracts of O. canum FT-IR analysis of chloroform extracts show the presence of some functional groups with corresponding intensity peaks O-H stretching or H bending for alcohols or phenols groups of active compounds,C-H stretching which was correspond to alkenes, N-H bend for 1º amines, C- C stretching was denoted the aromatic ring (Table 4). C-H(-CH2X) which is assumed alkyl halides groups of bioactive compounds then C- N stretching, which may indicates aliphatic amines. C-Cl stretching which may denoted for alkyl halides groups of component, C-H oop for aromatics groups of bioactive component, C-Cl stretching for corre- sponds to alkyl halides, C-Br stretching for alkyl halides groups and - C=C-H or CH bending denoted for alkynes (Figure 3). The results of FT- [Journal of Entomological and Acarological Research 2016; 48:4938] [page 349] Article Figure 1. Thin layer chromatography profile for chloroform extracts of O. canum. Figure 2. High-performance liquid chromatogram of chloroform extracts of O. canum. Table 3. High-performance liquid chromatography analysis of crude extracts of chloroform leaf extract of O. canum. Peak Ret. Tim Area Height Area% Height % 1. 3.482 19975 365 8.170 8.284 2. 5.459 4502 144 1.841 3.255 3. 9.960 133262 2438 54.504 55.264 4. 12.688 86759 1465 35.485 33.198 No n- co mm er cia l u se on ly [page 350] [Journal of Entomological and Acarological Research 2016; 48:4938] IR highlights the bioactive phenyl propanoid from O. sanctum (Upadhyaya et al., 2014) and crude extracts of O. bacillicum (Gabi Baba et al., 2012) which showed active functional groups. GC-MS analysis of chloroform extracts of O. canum GC-MS analysis of crude chloroform extracts of O. canum were identified as contain seventeen bioactive compounds (Figure 4 and 5). Among them, six (Bicyclo[2.2.1]heptan-2-one,1,7,7-trimethyl-,(IS), Azulene,1,2,3,3A,4,5,6,7-Octahydro-1,4-dimethyl-7-ci-methylethenyz)- ,[Ir-(1.alpha.,3A.beta,4 alp., methyl8,11,14-heptadeca-trienoate,2,6, 10,14,8, 22- Tetracosahexaene,2,6,10,15,19,23-hexamethyl(ALL-E)-, eicosane, hentriacontane)were considered as major and remaining minor compounds (caryophyllene,Z,Z-6,28-heptatriactontadien-2-one, Z,Z-6,28-Heptatriactontadien-2-ONE, Phytol, CIS-1-chloro-9-octadecene, 3,7,11,15-tetramethyl-2-hexadecen-1-OL, Hentriacontane, 5-Acetoxy methyl-2,6,10-trimrthyl-2,9-undecadien-6-OL, 2H-1-Benzopyran-6-OL, 3,4-Dihydro-2,5,7,8-Tetramethyl-2-(4,8,12-Trimethyltridecyl)- ,Acetate,[2R-[Gamma.-sitosterol) based on the retention time, peaks, molecular weight and percentage of area.Similar kind of work was done by several researchers with different lamiaceae plant species i.e., Stachys oblique (Harmandar et al., 1997), Origanum dictamnus, Teucrium poli- um and Lavandula vera (Proestos et al., 2006), Teucrium marum subsp. marum (Ricci et al., 2005), Nepeta argolica (Skaltsa et al., 2000) and Thymus comosus (Pavel et al., 2009) and supports the findings of the present study. Article Table 4. Fourier transform infrared spectroscopy analysis of chlorofom leaf extract of O. canum. S. No Peak range Types and functional group Bonding pattern 1 3377.12 Alcohols, phenols O-H str (s),H band (b) 2 2930.77 Alkanes C-H stretching 3 1572.21 1° Amines N-H (bend) 4 1403.82 Aromatics C-C (str) (in ring) 5 1260.71 Alkyl halides C-H (-CH2X) 6 1122.19 Aliphatic amines C-N (str) 7 1031.51 Aliphatic amines C-N (str) 8 923.13 Carboxylic acid O-H (bend) 9 831.98 Alkyl halides C-Cl (str) 10 757.23 Aromatics C-H ‘OOP’ 11 696.11 Alkyl halides C-Cl str 12 651.77 Alkyl halides C-Br str 13 618.90 Alkynes -C=C-H;CH bend Figure 3. Fourier transform infrared spectroscopy analysis of chloroform extract of O. canum. No n- co mm er cia l u se on ly [Journal of Entomological and Acarological Research 2016; 48:4938] [page 351] Article Figure 4. GC-MS analysis of chloroform extracts of O. canum. No n- co mm er cia l u se on ly [page 352] [Journal of Entomological and Acarological Research 2016; 48:4938] Conclusions The chloroform extracts of Ocimum canum showed highest mortal- ity against A. aegypti vector in all stages. Presence of potential phyto- constituents was identified using preliminary screening test and the functional groups of phytoconstituents were characterized by FT-IR and HPLC. The structural derivation of the identified compounds was car- ried out by GC-MS analysis. The overall results concluded that the chlo- roform extracts contain potential bioactive compounds for the mosqui- toes repellent, which can be useful for development of green based mosquitocidal agents in future for the control of vector borne diseases especially dengue fever. References ABBAS M.N., RANA S.A., UL-HASSAN M.M., RAN N., IQBAL M., 2013 - Phytochemical constituents of weeds: baseline study in mixed crop zone agro ecosystem. - Pak. J. Weed Sci. Res. 19: 231-238. 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