R. K. Harwansh et al. / Journal of Pharmacy Research 2009, 2(5),978-982 Research Article ISSN: 0974-6943 Available online through www.jpronline.info The medicinal as a potential source for agricultural crop protection against aphides (Brown Planthopper, Nilapurvata lugens; (Stal) (Hemiptera : Delphacidae) as pesticides. R. K. Harwansh*1, N. Shrivastava1 J. S. Dangi2 *1Department of pharmacy, Surguja University, Ambikapur-497001,Chhattisgarh, India. 2SLT Institute of Pharmaceutical Sciences; Guru Ghasidas University, Bilaspur-495001Chhattisgarh, India. Received on: 24-09-2008; Accepted on: 07-03-2009 ABSTRACT Methanol extract of c. swietenia of medicinal plant was tested for antifeedent, activity against third instars larva of cotton worm (spodoptera littoralis). And the methanol extract was also studied for the adulticidal activity and repellency test in red four beetles (tribolium castenium). The % mortality for adulticidal activity at different concentration (mg/ml) 1,2,3,4, 5 was recorded during the time interval 0 to 10 hour; 0%, 10%, 70%, 80%, 90%, 100%, respectively. LD50 and LD90 were 2mg/ml and 4mg/ml respectively. Statistical analysis of data the P- value is 0.0005, considered significant. Dunnet multiple comparision test the P-value is P<0.01, comparision test between ANOVA and Dunnet, the P-value is 0.0012, considered very significant. The antifeedent activity as % AI was noted according to the concentration (mg/ ml) 0,2,4,5, the LC50 was noted 0.414, 0.104, 0.081, and 0.016, respectively. The % repellency was noted according to the concentration (mg/ml) 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, between the time interval 0 to 10 hour; 0%, 0%, 10%, 20%, 30%, 40%, 60%, 70%, 80%, and 90%. The

LD50 and LD90 was 3.5mg/ml, 5mg/ml respectively. And repulsion test for stored product pest control management in the field of agriculture. The volatile oil from the leaves of c. swietenia was obtained via hydro distillation and was tested for pesticides activity against aphides brown plant hopper (Nilapurvata lugens) (stal) (Hemiptera: Delphacidae). And methanol extract of leaves of c. swietenia tested as adulticides against beetles act as natural insecticides.

Keywords: Chloroxylon swietenia; antifeedent; medicinal plant; red flour beetle; aphides; repellency test, pesticides, natural insecticides. INTRODUCTION According to the use of traditionally use of medicinal brown plant hopper (Nilapurvata lugens). Took hydrodistilled prod- lead to the development of new pesticides for crop protection against uct as essential oil through Clevenger apparatus. aphides (green/brown leaf hopper). Nowadays synthetic insecticides The essential oil applied into the eggs of aphides directly by or pesticides having a nature of chemical origin such as organo- spray in the crop of rice during the season of October-February be- phosphorus, carberyl, mercurial etc. compounds are mostly used for cause this season was life period of growth of plant hoppers depend- agricultural pest control and management so the insects developed ing upon the climatic conditions. In north India however, the popula- resistance against this chemical compounds so it was harmful for tion of this plant hopper becomes high in September–October. The environmental and human beings and ultimately leads to the toxicity adults remain most active from 10 to 320C. The females start laying and disease. So this why we have to choose a natural insecticides i.e. eggs within 3-10 days of their emergence and they deposit eggs in pyrethrines, azardirectines, nicotine’s, etc. having the potential source masses; by lacerating the parenchymal tissues. The number of eggs of insects killing and growth control. So one of this was a new agent per mass varies from 2-11 and female lays, on an average, 124 eggs (chloroxylon swietenia) having a potential activity against eggs of masses1. insects, larva, beetles insect and pests. And aphides as same as syn- The brown plant hopper is the most destructive pest of rice thetic insecticides. in south and south East Asia, China, Japan and Korea. In India, it has For this purposes we have to took out extract from plant as become very serious on the high yielding verities of paddy in many methanolic and hydrodistilled for different activity tested adulticidal, states including U.P, M.P., C.G., West Bengal; A.P. And Tamil Nadu. larvicidal, pesticidal and repellency activity respectively against red Both adults and nymph feed on paddy, sugarcane and grasses by flour beetles (tribolium castaneum) and cotton worm (spodoptera sucking the cell sap. The brownish adults with brown eyes are 3.5- ltitoralis). For crop protection and management against aphides 4.5mm in length. The nymph is brownish blacked in colour and have grayish-blue eyes1. *Corresponding author. Most of the white backed planthopper, green hopper, Rice Tel.: + 91-9754806921 leaf hopper, zig-zag leafhopper having sensitive for the product of c. Telefax: +91- swietenia as previously studied on essential oil composition of c. E-mail: [email protected] swietenia that was pregejerene, geijerene, germacrene-D, having the potential source for adulticidal, larvicidal and insecticidal activity2,3.

Journal of Pharmacy Research Vol.2.Issue 5.May 2009 978 R. K. Harwansh et al. / Journal of Pharmacy Research 2009, 2(5),978-982 According to this we have to point out the control of pest in 3.2 Antifeedent activity against spodoptera littoralis the area of agricultural field chooses this plant for control of pest in a Antifeedent activity was carried out using the filter paper crop of cereals viz, jawar, bajra, ragi, wheat and mainly rice in the disc choice method. The fresh green leaves of vegetables were used agriculture area of Chhattisgarh and M.P. as pasted form. The crude extract of compounds at different concen- trations was applied on to one side of disc the nicotine was used as 2. MATERIALS AND METHODS reference compound. Filter paper disc treated with methanol alone 2.1 Insects used as control. A red flour beetles (tribolium castaneum) was naturally cul- Freshly moulted 3rd instar larvae were taken and starved for tivated in flour of wheat in climatic conditions during the period of 25 4hr prior to testing. One larvae was placed in each Petri plate contain- days. Kept this in nutritive media until the used4. ing filter paper disc with small pieces of wet control to prevent desic- 2.2Larvae cation4. The procedures were performed in triplicate. The consump- Experiments were carried out on 3rd instar larvae (10-12mg) of tion of filter paper (green leaves pasted) was noted after 10 hr in both spodoptera littoralis (Biosdual); (Lepidoptera : Noctuide), kept on control and test disc. An antifeedent activity index (AI) was calcu- synthetic diet5. The larvae were selected under light, dark photoperi- lated from the formula8. odic form at room temperature. Obtained from sericulture department Formula: as natural resources. AI (%) = C-T/C+T×100 Where,C = weight of control filter paper with green leaf 2.3 Aphides paste consumed.T = weight of treated filter paper consumed. A brown plant hopper (nilapurvata lugens), (stal), (hemiptera : delphacidae)was obtained from the natural sources of agricultures. 3.3 Repellency test on red flour beetles (tribolium castaneum) or The eggs and nymph were chooses under climatic condition. Arena ring test 2.4 Plants Chloroxylon swietenia belonging to family is a It may be carried out on red flour beetles as test insects. A medicinal plant having synonyms in; English known as Indian satin filter paper disc was cut in two halves (test and control). The test disc wood, Yellow wood, East Indian satin wood, Baruta and Ceylon satin was treated with methanolic extract of 0.5-5mg/ml concentration and wood but in, Hindi known as Bhirra, Girya, Karwar and Bharhul. Plant dried. The control disc was treated with methanol, dried and both leaves of c. swietenia obtained from forest of Kataravan District – discs (halves) were joined together. About 10 adult insects were re- Janjgir Champa, Chhattisgarh. leased per petridishes at centre of disc and covered with glass ring. A specimen of plant was identified by Pankaj Oudhia, Bota- The distribution of insects in two halves was monitored for 10 hr4. nist; Surguja campus Chhattisgarh and we have also studied a num- ber of books and site of www.enwikipedia.org.com. 4. RESULTS 2.5 Extraction Represented in table and graph 1-3 respectively. Extraction of leaves of medicinal plant c. swietenia in the powdered form by soxhlet assembly. Took 50g of powdered crude 5. DISCUSSION drug macerated with solvent. In the next step took 250ml of methanol In the present study, the plant c. swietenia was chooses in round bottom flask of soxhlet apparatus. It was processing for primarily on the basis of ethnobotanical survey in the region of about 24hr at 550C. Finally take out the concentrated form of extract by Kataravan Distt. Janjgir-Champa; Chhattisgarh according to its uses evaporation on water bath to dryness and obtained crude dried ex- as pesticides in traditional medicine. As previously studies on the tract. composition of hydrodistilled product of volatile oil of c. swietenia 2.6 Hydrodistillation evaluated by topical application on the insects or larvae of cotton The leaves were dried in cool place and hydrodistilled in worm; it provides the acute toxicity effect against insect3. It is well distillation unit for 4hr. The distilled product was dried over anhy- known that various volatile oil and their constituents such as geijerene drous sodium sulphate and stored in refrigerator until further used3. and pegeijerene were found in considerably large quantities in the volatile oil of c. swietenia3 and no activity concerned to these sub- 3. BIOASSAY stances has so for been present on the rice or cereal crop of agricul- 3.1 Adulticidal activity on red flour beetles (tribolium castaneum) tural insects for protection, control and management. The aphides A film of methanolic extract of c. swietenia at different con- brown planthopper destroy the crop of rice, wheat etc. And its paddy centration (2, 4, and 5mg/ml) was prepared in the petridishes (10cm by sucking the cell sap from the leaves which turn yellow and ulti- dm). Then film was dried by evaporation of solvent in warm room mately destroyed the whole plants of rice this is infestation for crop in temperature. Adults of red flour beetles (tribolium castaneum) were the field of agriculture1. So we have to choose a natural pesticides put in to the petridishes having two treatments first one was control over their harmful effect or adverse effect on agroecological system treatment and second one was extract treatment. Then covered and are well known. maintained at 270C, and 60% RH for 10hr. Finally mortality was noted4. The huydrodistilled and methanolic compound of volatile oil of c. This method was performed in triplicate. swietenia provides topical effect on insects. The provides typical neurotoxic symptoms found in insects, including tremors, lack of Journal of Pharmacy Research Vol.2.Issue 5.May 2009 979 R. K. Harwansh et al. / Journal of Pharmacy Research 2009, 2(5),978-982

Graph1. Adulticidal activity Graph2. Antifeedent activity

Triboleum Adulticidal activity on Antifeedent activity against Spodoptera casteneum litoralis 150 % 0.5 m 100 0.4 or Series1 0.3 tal 50 Series1 0.2 ity 0 0.1

0 1 2 3 4 5 6 % Antifeedent activity 0 Concentration 0 2 4 6 (mg/ml) Conc.(mg/ml)

Graph3. Repellency test

Repellency test against Triboleum casteneum 100

80

60

40 Series1 20

% Repellency 0 0 1 2 3 4 5 6 -20 Conc.(mg/ml)

Table1:- Adulticidal activity

S. No. Time (hr) Conc.(mg/ml) No. of Beetles used Mortality mean ± SD % Mortality

1 0-1 0 10 0±0.0 0 2 1-2 1 10 1±0.08 10 3 2-4 2 10 7±0.1 70 4 4-6 3 10 8±0.3 80 5 6-8 4 10 9±0.3 90 6 8-10 5 10 10±0.4 100 Table2:- Antifeedent activity S. Time Conc Wt. of each Dm of each T (mg) C(mg) % of anti-feedent No. (hr) .(mg/ml) filter paper(mg) filter paper (cm) (mg) 1 0-4 0 500 10.5 605 600 0.414 2 4-6 2 500 10.5 610 608 0.164 3 6-8 4 500 10.5 618 617 0.081 4 8-10 5 500 10.5 620 618 0.016

Journal of Pharmacy Research Vol.2.Issue 5.May 2009 980 R. K. Harwansh et al. / Journal of Pharmacy Research 2009, 2(5),978-982 Table3:- Repellency test

S. No. Time (hr) Conc.(mg/ml) No. of Insects / No. of insects % Repellency Petridish repelled mean ± SD 1 0-1 0.5 10 0±0.00 0 2 1-2 1.0 10 0±0.00 0 3 2-3 1.5 10 2±0.05 10 4 3-4 2.0 10 3±0.09 20 5 4-5 2.5 10 4±0.12 30 6 5-6 3.0 10 5±0.15 40 7 6-7 3.5 10 6±0.2 60 8 7-8 4.0 10 7±0.25 70 9 8-9 4.5 10 8±0.26 80 10 9-10 5.0 10 9±0.29 90 coordination, finally causes paralysis death are assume due to the 6. CONCLUSION blockage of neurotransmitters by the volatile oil and related com- 3 pounds. According to the previously studies involving toxicity of The goal of my present work indicates that the volatile oils conventional insecticides, geijerene is seven time less toxic then the and crude product of methanolic extract of c. swietenia provides pes- standard synthetic organophosphate, monocrotophos, 35 times less toxic than the organochloride, endosulphan and 81 times less toxic ticidal activities comparing with commercial pesticides compared to the carbonates methomyl3. monochrotophos and nicotine. Almost, these results may lead to the Topical application to the oils and the methanolic crude ex- development of newer and more selective natural compounds. tract delayed larval development via decreased rate of growth.Further the volatile oil and crude extract were also experimented for their effi- 7. ACKNOWLEDGEMENT ciency as bioassay on insects as adulticidal, antifeedent against red flour beetles (tribolium castaneum) and spodoptera littoralis respec- I grateful to Dr. Pankaj Oudhia, Botanist of Surguja, tively. And also performed repellency test on insect’s tribolium Chhattisgarh for identification of plant leaves of Chloroxylon castaneum. And finally experimented on crop insect’s brown plant swietenia. My thanks extend to my family especially my father and 1 hopper (Nilapurvata lugens) (stal), (Hemiptera : Delphacidae) . Both mother for their motivation in every stage of my studies. And my compound oils and crude extract exhibited significant activity such as thanks also extends to my supervisor; Prof. (Dr) J. S. Dangi, teachers; adulticidal and feeding inhibition, repellency and pesticidals. The % mortality for adulticidal activity at different concentration (mg/ Dr Alpana Ram, Mr. S. H. Bodake SLT Institute of Pharmaceutical ml) of MECS (or methanol extract of c. swietenia) 1,2,3,4, 5 was re- Sciences, Guru Ghasidas University, Bilaspur; Chhattisgarh. I would corded during the time interval 0 to 10 hour; 0%, 10%, 70%, 80%, 90%, like to express my regards to my class fellow Mr. Rohitas Deshmukh for their kind suggestion cooperation and appreciation in initial stage 100%, respectively. LD50 and LD90 were 2mg/ml and 4mg/ml respec- tively. Statistical analysis of data (According the software of statis- of my studies. tics Graphpad-Instat) the P-value is 0.0005, considered significant, variation among column means is significantly greater than by chance. REFERENCES Dunnet multiple comparision test the P-value is P<0.01, comparision test between ANOVA and Dunnet, the P-value is 0.0012, considered 1. Alwal A.S. & Dhaliwal G.S. “Agriculture pest of south Asia & very significant. Effective matching results in significant variation their management” Kalyani Publisher, Ludhiyana, New Delhi, among means, with these data the matching appears to be effective. Noida (U.P), Hyderbad, Chennai, kakkata, Cuttak, Edition The antifeedent activity as % AI was noted according to the (2002), p 179. 2. Kiran S.R., Devi P.S. “Evaluation of mosquitocidal activity of concentration (mg/ml) 0,2,4,5, the LC50 was noted 0.414, 0.104, 0.081, and 0.016, respectively. The statistical treatment of data Paired t-test essential oil & sesquiterpenes from leaves of Chloroxylon swieteria DC.” Parasitol Res (2004), 101; p 413-418. (or one-tail P-value), the one-tail P-value is 0.0298, considered signifi- 3. Kiran S.R. , Devi P.S. & Reddy K.J. “Bioactivity of essential cant and T = 2.060 with 3 degree of freedom. oils & sesquiterpenes of Chloroxylon swieteria DC, against The % repellency was noted according to the concentration Helicoverpa armigera” current science vol.- 93 No. 4, 23 (mg/ml) 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, between the time August 2007. interval 0 to 10 hour; 0%, 0%, 10%, 20%, 30%, 40%, 60%, 70%, 80%, 4. Kokate, C.K., Purohit A. P. and Gokhle S.B. “Pharmacognosy” st and 90%. The LD50 and LD90 was 3.5mg/ml, 5mg/ml respectively. The Nirali Prkshan, 21 edition pune, p 129-130. statistical analysis of data Paired t-test (or one-tail P-value), one-tail 5. Pavela R. “Insecticidal activity of certain medicinal plants” P-value is 0.0001, considered extremely significant; t = 6.000 with 9 Fitoterapia 75 (2004) p 745-749. degree of freedom. Effective pairing results in a significant correlation 6. Hooker J.D. “Systema of Botany” Descriptive of Analytical between the columns, with these data, the pairing (or matching) ap- in two parts Part I- Outline of organographiy, “anatomy & physiology” Part II- Descriptive & illustration of the order. pears to be effective. Reprint 1986, printed & published by –“International books and periodical supply service 240/5 Prakashan, Gupta road, kawal bagh, New Deldi-110005, p 335. Journal of Pharmacy Research Vol.2.Issue 5.May 2009 981 R. K. Harwansh et al. / Journal of Pharmacy Research 2009, 2(5),978-982 7. Kirtikar K.R. & Basu E.D. “Indian Medicinal Plant with Illus- 8. Han M.K., Kim S-II, Ahn Y.J.” Insecticidal & Antifeedent tration”, oriented ennterprises, Published & Printed by Mr. activities of Medicinal plant extract against Attagenus Devendera singh for oriental enterprises, Rajpur, Dehradun - unicolor japonious (Coleopatera : Dermestidae)” Journal of 24801,Uttranchal, India, Vol-I (2001)Plot No.-231 stored products Research 42 (2006) p 15,22 Source of support: Nil, Conflict of interest: None Declared

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