Pak. Entomol. Vol. 31, No.

1, 2009 EFFECT OF NEEM DERIVATIVES ON INFESTATION, SETTLING AND OVIPOSITION OF MELON FRUIT FLY (BACTROCERA CUCURBITAE COQ.) (TEPHRITIDAE: DIPETRA) Masood Khan Khattak, Mohammad Mamoon-ur Rashid and Khalid Abdullah* Department of Entomology, Faculty of Agriculture, Gomal University, Dera Ismail Khan. * Agriculture Research Institute, Dera Ismail Khan. ABSTRACT In order to evaluate the effect of neem oil and neem seed water extract on the infestation, settling and oviposition of melon fruit fly, (Bactrocera cucurbitae Coq.), field as well as laboratory trials were conducted on Bukhara variety of melon at Hathala, D.I.Khan. Neem oil and neem seed extract were prepared from fresh neem seeds collected from the area and different concentrations of 1, 1.5 and 2% and 1, 2 and 3% were made in water to spray in the fields, respectively. The concentrations viz., 1000, 5000 and 10,000 ppm, of both neem oil and neem seed extract were used in laboratory tests. In the field trials, neem oil and neem seed water extract at all tested concentrations reduced the fruit fly infestation. Significantly less number of pupae were recovered from the randomly selected fruits in the treated plots as compared to control. The effect of both neem derivatives was dose dependent. However, adult emergence of the flies was not affected at any level of concentration. In laboratory test, both neem oil and neem seed water extract at 10,000 ppm adversely affected the settling of melonfruit fly; as 1.7 flies and 1.7 flies on fruits treated with 10,000 ppm of neem oil and 10,000 ppm of neem seed water extract, respectively, were significantly lower than 6.3 and 7.3 flies in their respective control. Similarly pupal recovery at all tested concentrations of neem oil and neem seed water extract were statistically lower than that in their respective control. Key Words: neem derivatives, melon fruit fly, deterrent, repellent, anti-ovipositional. INTRODUCTION Muskmelon (Cucumis melo L.) is an important cash crop of the poor farmer, especially in the Barani (rainfed) areas in Pakistan. Insect pests cause considerable losses to this crop. Among these insect pests fruit flies (Diptera: Tephritidae) are of vital importance. About 4000 species of fruit flies has been identified in the world, out of which 450 cause considerable damage to fruits and vegetables (Knight, 2003). Fruit fly can live up to three months as adult and has a capability of laying up to 1000 eggs during its life time (Irshad and Jilani, 2003). In Pakistan, there are 11 fruit flies species. Bactrocera zonata Saund and B. dorsalis Hend. attack mango, citrus, guava, melon, ber and date palm. Myiopardalis pardaliva Bigot attacks melon (Abdullah and Latif, 2001; Abdullah et al., 2002). Several management techniques are being applied to overcome this notorious insect pest because three of its life stages are hidden and the only adult stage is the usual target of the pest control activities. The local farmers because of their limited approach and lack of knowledge use conventional insecticides for its control without knowing the ill effects of these chemicals on humanity. Alternative botanical insecticides can play important role as a component of IPM strategies against fruit flies. Among botanicals, neem has great potential for commercial exploitation. Neem has an advantage over synthetic insecticides in that it has no or less toxicity to man, other animals and useful insects (Schumutterrer, 1985). Neem products affect insect vigor, longevity and fecundity (Arora and Dhaliwal, 1997). The triterpenoids, meliantriol, salannin and azadirachtin mainly occurring in the neem seed act as antifeedant, ovipositional, deterrence, disturb insect growth and development (Prakash and Rao, 1994). Extracts from neem has proved good repellents against many insect pests. Neem extracts showed strong repellent activities against mosquitoes when mixed at 0.01% and 1.0% with kerosene oil and burnt in lamps (Sharma and Ansari, 1994). Khattak et al. (2001)

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Pak. Entomol. Vol. 31, No.1, 2009 observed that neem oil, at 1000 ppm and 10,000 ppm, was good repellent against maize weevil (Sitophillus zeamais Motsch.) and has no effect on its parasitoid Anisopteromalus calandrae Howard. Maize weevil settled significantly in lower number and also laid fewer eggs on corn kernels as compared to the untreated ones. The neem based insecticides in artificial diet resulted decrease in pupal formation and subsequent adult emergence of the western cherry fruit fly (Randen et al. 1998). Neem extracts at 0.5% significantly reduced 81.8%, 75.11% and 82.2% infestation of fruit flies in mango, muskmelon and guava, respectively as compared to that in the control (Annonymous, 2003). In this paper, Effect of neem derivatives on the infestation, settling and oviposition of melon fruit fly, (Bactrocera cucurbitae Coq.) has been examined. MATERIALS AND METHODS Preparation of neem oil concentrations 1 year old neem seed was collected from the local farmers and the oil from these seeds was extracted with oil extractor. Different concentrations of the crude neem oil were prepared in water to be used in these trials. Preparation of neem water extracts 2 Kg dried neem seed collected from the local farmers was ground into a fine powder. This powder was tied in a cotton cloth in the form of a bag and dipped in five liter of water at 80C for 16 hours. In this way concentrated solution of 20 % was obtained which was diluted to 1, 2, and 3% for spray in the trials. Field experiment An experiment was conducted at farmers field in Hathala, D.I.Khan, where heavy attack of melon fruit fly has been reported, to determine the effect of neem oil and neem seed water extract at different level of concentrations on the infestation of melon fruit fly. The seeds of melon variety (Bukhara) used in this experiment was obtained from the local market. The experimental layout was Randomized Complete Block Design (RCBD), with seven treatments including a control; each treatment was replicated three times on a plot size of 24 m2 with 45 cm (p x p distance) and 2 m as R x R distance. All other standard agronomic practices were given at proper time. Laboratory trials This experiment was conducted at Fruit Flies Experimental Laboratory, Agriculture Research Institute, D. I. Khan, to investigate the effect of neem oil and neem seed water extract at different concentrations on settling and oviposition of melon fruit fly (B. cucurbitae). The number of pupae recovered in the experiment was considered as the effect of neem derivatives. Three concentrations of each neem derivative, i.e., 1000, 5000 and 10000 in acetone were prepared. Two melon fruits almost similar in size and color were collected from the field where no pesticides were applied during the season and the fruits were protected from the infestation of flies through net after thoroughly washing and air drying. The fruits were dipped in each cocnetraion for 1 minute. They were kept in open air for 10 minutes to completely evaporate the solvent. The fruits were marked for the treatment. Control fruits were dipped in acetone only and then air dried. The treated and untreated fruits were placed in alternating fashion and at equal distance to each other in cages (measuring 90 x 90 x 90 cm3) with glass on four sides and fine mesh on the top for aeration. Ten female flies 15 days old were collected from the lab-reared population and released in each cage. After each 30 minutes, the settling of flies on each fruit was recorded. Those flies were excluded from the data which did not respond i.e. they settled neither on treated fruits nor on untreated ones. After 24 hours, fruits were taken out of the cages and placed separately in plastic jars (1000 ml The neem derivatives were sprayed with 10 liters knap sack hand operated sprayer. The control plot was only sprayed with water only. The plants were sprayed after fruit setting. The spray was repeated after 15 days. Ten days after second spray, five (5) fruits were randomly selected from each treatment and placed separately in rearing cages with saw dust or soil at the bottom. The difference in pupal recovery and adult emergence were kept a parameter for the effect of neem derivatives on the infestation of melon fruit fly. The soil in each cage was sieved and pupae were counted after 12 days and placed separately for adult emergence. The data for adult emergence were recorded after 10 days of the pupal formation. Thus the final data were analyzed with Analysis of Variance (ANOVA) and means were separated using Least Significant Different (LSD) test.

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Pak. Entomol. Vol. 31, No.1, 2009 capacity) containing sawdust at the bottom for pupal recovery. After eleven days, saw dust was sieved and pupae were collected and counted. The pupae were again placed in 10cm diameter petri-dish for adult emergence. After 10 days the number of adult flies emerged were recorded. Each treatment in the experiment was replicated three times and the data obtained was analyzed using t Test software and means were separated by DMRT for their comparison. RESULTS Field trials As the fruit flies lay eggs beneath the skin of the fruit and counting of eggs is not possible, therefore, the oviposition response of the test insect would either be noted by counting the larvae in the infested fruit by dissecting them or by counting the number of pupae recovered in each treatment. In field experiment, significantly low number of pupae was recovered from the fruits treated with neem derivatives as compared to that in the control. Maximum (54.67) number of pupae was recovered from the untreated fruits (Table 1). Table 1. Effect of neem derivatives on the infestation of melon fruit fly Treatments Infestation data No. of pupae % adults emerged Neem oil recovered 1% 37.67c 88.48 ns 1.5% 22.67e 89.86 2% 8.33f 83.79 Neem seed water extract 1% 48.33b 91.13 2% 32.33d 90.84 3% 18.33e 83.77 Control 54.67a 91.56 LSD 5.33 7.92
Each value is a mean of three replications. Means sharing the same letters in a column are not significantly different from each other at = 0.05

neem oil, respectively. Similarly significantly lower (18.33) number of pupae were recovered from fruits treated with 3% neem seed water extract than 48.33 and 32.33 pupae recovered from fruits treated with 1% and 2% neem seed water extract, respectively. None of the neem derivatives affected adult emergence of the fruit fly. Statistically similar percentage of adults emerged from the pupae recovered from all treatments as compared to that in the control (Table 1). Laboratory trials When fruit flies were given a chance of distribution on musk melon fruits treated with neem derivatives and untreated in a closed arena, significantly lower number of flies (1.7) settled on fruits treated with 10000 ppm of neem oil and neem seed water extract as compared to their respective controls (Table 2). Other concentrations of neem oil and neem seed water extract did not repel the fruit flies; as 3.0, 2.3 flies and 3.7 and 3.3 flies on fruits treated with 1000 ppm and 5000 ppm of neem oil and neem seed water extract, respectively, were statistically similar to the number of flies in their respective controls. In the present trials, all concentrations either of the neem oil or neem seed water extract adversely affected the oviposition of the test insect as 11.0%, 7.0% and 5.0% pupae recovered in the neem oil at 1000 ppm, 5000 ppm and 10000 ppm, respectively which were significantly lower than the pupae in their respective controls (Table 2). Similarly the pupae recovered in the neem seed water extract concentrations were also statistically lower than the pupae in their respective controls. DISCUSSION Neem oil and neem seed water extract at 1%, 2% and 3% reduced the fruit fly percent infestation; as the pupae recovered in each treatment was significantly fewer as compared to that in the control; however, percent adult emergence was not affected by any concentration. In laboratory test, settling of the flies was only affected with 10,000 ppm neem oil and neem seed water extract but significantly lower pupae were recovered in all tested concentrations than in their respective controls. Other research workers also obtained identical results with the neem compounds and other botanical insecticides against fruit flies as well as other insect pests. Valencia- Botin et al.

The effect of neem derivatives on the test insect was dose dependant. The higher concentration of neem oil and neem seed water extract resulted into fewer number of pupae than the pupae in the lower concentrations; as 8.33 pupae recovered from the fruits treated with 2% neem oil which were significantly lower than 37.67 and 22.67 pupae recovered from the fruits treated with 1% and 1.5%

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Pak. Entomol. Vol. 31, No.1, 2009 (2004) suggested that neem could be incorporated within strategies of integrated pest management in citrus; as they observed significant repellency in the oviposition of the Mexican fruit fly at 3 and 5% aqueous neem extract and 4.5% neem oil treatment. Akhtar et al. (2004) observed that lower number (0.94) Bactrocera zonata adults settled on frtuits treated with sweetflag pertroleum ether extract followed by (1.00) flies on neem acetone extract and (1.00) flies on sweetflag extract. The number of pupae and adults obtained from fruits decreased with increase in the extract concentration. In a series of experiments, Singh (2003) demonstrated that neem extracts can be effectively used as an excellent alternative to synthetic insecticides. Table 2. Effect of neem derivatives on settling and oviposition of melon fruit fly Treatment Settling response Pupal recovery Neem oil 1000 ppm 3.0a 11.0a Control 5.7a 28.3b t value -4.00 -8.54 5000 ppm 2.3a 7.09 Control 6.0a 30.0b T value -4.15 -6.07 10000 ppm 1.7b 5.0a Control 6.3a 33.7b t value -14.00 -17.2 Neem seed water extract 1000 ppm 3.7a 17.7a Control 5.3a 27.7b t value -2.50 -8.66 5000 ppm 3.3a 12.3a Control 5.3a 24.3b t value -1.73 -5.19 10000 ppm 1.7b 4.0a Control 7.3a 36.3 t value -4.71 -11.12
Paired values within column followed by the same letters are not significantly different at = 0.05. Each pair represents the choice offered in an arena.

Jilani et al. (2006) found that petroleum ether extract of Valariana officianalis at 250 ppm, 500 ppm and 1000 ppm in diet significantly prohibited the pupal formation; however, only 1000 ppm and 500 ppm extracts in diet reduced adult emergence significantly.

Khattak et al. (2006) noted that significantly lower number of peach fruit flies settled on guava fruits treated with 1% extract of Peganum harmala, Saussuria lappa and Valariana officianalis. Pupal formation and progeny adult were totally prohibited in 1% petroleum ether extract of S. lappa. In the field experiment. In comparative studies, Gyi et al. (2003) observed that although, nimbecidine 5ml L-1 remained with highest (37.3%) infestation of Carpimoya vesuviana infesting ber as compared to 13.7% infestation with lambada-cyhalothrin and 15% infestation with beta-cyfluthrin, but was significantly lower than the control. Vincenzo et al. (1999) determined than commercial neem formulation significantly reduced the fecundity of Ceratitis capitata (Diptera: Tephritidae). The number of eggs laid decreased with increasing concentration of azadirachtin in diet. Singh et al. (2000) compared econeem with many other synthetic insecticides at their recommended doses against ber fruit fly. They found that all the treatments reduced the infestation of the test insect compared to the control. Jaferry (2006) observed significantly improved activity of the bait (protein hydrolysate) application when mixed with neem (Azadirachta indica) extract as compared to synthetic insecticide (Diptrex 80SP). The pupal recovery from the harvested fruits was low in neem extract treatments followed by synthetic insecticide and untreated check. The trend of pupal recovery in the neem extract treatments decreased with the increase in neem concentrations. All the neem oil treatments had significantly fewer flies settled on the fruits. Neem oil repelled the flies and thus inhibited the oviposition. The trend of flies settlement decreased with each increment in the neem oil concentration. As the test insect is very much odor conscious, the low pupal recovery in all neem oil treatments may be because of it high repellent, antovipositional and deterrent quality. From the results, it is evident that both neem and protein hydrolysate have broad scope in organic agriculture, match with many countries export demands and thus are best fitted in the WTO regulations. Neem oil treatments of fruits have significantly lower infestation then untreated fruits. This lower infestation in the dropped fruit clearly showed an ovipositional deterrent effect of the neem oil as reported by many researchers for different pest species in the world (Nadia and Schmidt, 1992; Nadia et al., 1995).

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Pak. Entomol. Vol. 31, No.1, 2009 ACKNOWLEDGEMENTS The financial support of Higher Education Commission Islamabad for the studies is highly appreciated and acknowledged. REFERENCES Abdullah, K. and A. Latif, 2001. Studies on baits and dust formulations of insecticides against fruit fly (Diptera: Tephritidae) on melon (Cucumis melo) under semi arid conditions of Dera Ismail Khan. Pak. J. Biol. Sci., 4 (4): 334-335. Abdullah, K., M. Akram and A. A. Alizai, 2002. Non-traditional control of fruit flies in guava orchards in D. I. Khan. Pak. J. Agri. Res., 17(2): 71-74. Akhtar, N., G. Jilani, R. Mahmood, M. Ashfaq and J. Iqbal, 2004. Effect of plant derivatives on settling response and fecundity of peach fruit fly (Bactrocera zonata) (Saund.). Sarhad J. Agric., 20 (2): 269-274. Anonymous, 2003. Integrated Management of Fruit Flies. pp. 1-51. Arora, R. and G. S. Dhaliwal, 1994. Botanical pesticides in insect pest management. In : Dhaliwal, G. S. and B. D. Kansal (eds.). Management of Agricultural pollution in India. Commonwealth Publ., New Delhi, India, pp. 213-245. Gyi, M. M., O. P. Lal, A. K. Dikshit and V. P. Sharma, 2003. Efficacy of insecticides for controlling Ber fruit fly. Ann. Pl. Prot. Sci., India, 11 (1): 152-153. Irshad, M. and G. Jilani, 2003. Management of fruit fly in Pakistan. Pest Management. Res. Prog. NARC, Islamabad, Pakistan. 46 pp. Jaffery, A. A., 2006. Bionomics and management studies of fruit fly, Bactrocera spp. (Diptera: Tephritidae) infesting Guava. M. Sc. (Hons.) Thesis, Fac. Agri. Gomal University, 32 pp. Jilani, G., M. K. Khattak and M. F. Shahzad, 2006. Toxic and growth regulating effect of ethanol extract and petroleum ether extract of Valariana officianalis L. against Bactrocera zonata (Saund.). Pak. Entomol., 28 (1): 11-14. Khattak, M. K., A. B. Borce and B. Dover, 2001. Comparative effects of neem or mineral oil on maize weevil, Sitophilus zeamais Motsch. and its parasitoid, Anisopteromalus calandrae (Howard). Online J. Biol. Sci., 1(5): 378-381. Khattak, M. K., M. F. Shahzad and G. Jilani, 2006. Effect of different extracts of Harmal (Peganum harmala L.), Rhizome of kuth (Saussurea lappa C. B. Clarke) and Balcher (Valariana officianalis L.) on the settling and growth of Peach fruit fly (Bactrocera zonata Saunders). Pak. Entomol., 28 (1): 15-18. Knight, S. A., 2003. Invasive species and pest management (ISPM): fruit fly program
information.http://www.aphis.usda.gov/ppq/ispm/ff.

Nadia Z. D., A. A. Barakat, E. F. Abdalla, H. E. ElMetwally and A. M. E. Abd El Salam, 1995. Evaluation of two neem seed kernel extracts against Liriomyza trifolii (Burg.) (Dipt. Agromyzidae). J. Pest Sci., 68 (2): 39-41. Nadia, Z. D. and G. H. Schmidt, 1992. Efficacy of Neem-Azal S and Margosan-O against the bean aphid, Aphis fabae Scop. J. Pest Sci., 65 (4): 75-79. Prakash, A. and J. Rao, 1997. Botanical pesticides in agriculture. CRC Press.USA. Randen, E. J., B. D. Roiberg and E. J. Randen, 1998. Effect of a neem (Azadirachta indica) based insecticide on oviposition deterrence, survival, behavior and reproduction of adult western cherry fruit fly (Diptera: Tephritidae), J. econ. Ento., 91 (1): 123-31. Schmutterer, H. 1985. Which insect pests can be controlled by application of neem seed kernel extracts under field conditions. Z. Ang. Ent., 100: 468-475. Sharma, V. P. and M. A. Ansari, 1994. Personnel protection from mosquitoes (Diptera: Culicidae) by burning neem oil in kerosene. J. med. Entomol., 31: 505-507. Singh, S., 2003. Effect of aqueous extract of neem seed kernel and azadirachtin on fecundity, fertility and post embryonic development of melon fruit fly, Bactrocera cucurbitae and B. dorsalis (Diptera: Tephritidae). J. Appl. Entomol., 27 (9/10): 540-547. Singh, S., R. N. Gupta, B. K. Awasthi and R. A. Verma, 2000. Effective control of Ber fruit fly , Carpomiya vesuviana by insecticidal scheduale. Indian J. Entomol., 62 (2): 171-174. Valencia-Botin, A. J., N. Bautista-Martinez and J. A. Lopez-Buenfil, 2004. Use of neem (Azadirachta indica A Juss) aqueous extract on the oviposition of Mexican fruit fly, (Anastrepha ludens), Loew) (Diptera: Tephritidae) in Valencia orange. Sanidad vegetal, Cludad, de La Habana, Cuba. 8 (4): 57-59. Vincenzo, D. I., C. Massino, N. Paola and D. Romano, 1999. Effect of neem compounds on the fecundity and longevity of Ceratitis capitata (Diptera: Tephritidae). J. Econ. Entomol., 92(1): 76-82.

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