Insecticidal activity of Poppy (Papaver somniferum L.) seed oil against cowpea weevil (Callosobruchus maculatus F.

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1 Payam Nour University From the SelectedWorks of Adel Khashaveh Spring May 9, 2013 Insecticidal activity of Poppy (Papaver somniferum L.) seed oil against cowpea weevil (Callosobruchus maculatus F.) in stored cowpea Adel Khashaveh Available at:

2 This article was downloaded by: [Adel Khashaveh] On: 09 May 2013, At: 08:40 Publisher: Taylor & Francis Informa Ltd Registered in England and Wales Registered Number: Registered office: Mortimer House, Mortimer Street, London W1T 3JH, UK Archives Of Phytopathology And Plant Protection Publication details, including instructions for authors and subscription information: Insecticidal activity of Poppy (Papaver somniferum L.) seed oil against cowpea weevil (Callosobruchus maculatus F.) in stored cowpea Hamid Sakenin Chelav a & Adel Khashaveh b a Department of Plant Protection, Qaemshahr Branch, Islamic Azad University, Qaemshahr, Iran b Young Researchers Club, Qaemshahr Branch, Islamic Azad University, Qaemshahr, Iran Published online: 09 May To cite this article: Hamid Sakenin Chelav & Adel Khashaveh (2013): Insecticidal activity of Poppy (Papaver somniferum L.) seed oil against cowpea weevil (Callosobruchus maculatus F.) in stored cowpea, Archives Of Phytopathology And Plant Protection, DOI: / To link to this article: PLEASE SCROLL DOWN FOR ARTICLE Full terms and conditions of use: This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. The publisher does not give any warranty express or implied or make any representation that the contents will be complete or accurate or up to date. The accuracy of any instructions, formulae, and drug doses should be independently verified with primary sources. The publisher shall not be liable for any loss, actions, claims, proceedings, demand, or costs or damages whatsoever or howsoever caused arising directly or indirectly in connection with or arising out of the use of this material.

3 Archives of Phytopathology and Plant Protection, Insecticidal activity of Poppy (Papaver somniferum L.) seed oil against cowpea weevil (Callosobruchus maculatus F.) in stored cowpea Hamid Sakenin Chelav a and Adel Khashaveh b * a Department of Plant Protection, Qaemshahr Branch, Islamic Azad University, Qaemshahr, Iran; b Young Researchers Club, Qaemshahr Branch, Islamic Azad University, Qaemshahr, Iran (Received 23 March 2013; final version received 30 March 2013) In this study, the insecticidal effect of Poppy (Papaver somniferum) seed oil was investigated against Callosobruchus maculatus (Bruchidae) in cowpea, at different concentrations and exposure time. The Poppy seed oil at 2, 4, 6, 8 and 10 ml/kg was tested against adults of C. maculatus, and the mortality was counted after 24, 48 and 72 h of exposure. All tests were conducted at C and 65 ± 5% r.h. The experiments were carried out based on the factorial experiment by randomized complete design with four replications. Twenty-five insects with 0 24 h old were used for each replication. The effect of Poppy seed oil on the reduction of emergence of insects in next generation (F 1 ) was also assayed. The results of experiments indicate the significant differences between concentrations and exposure time (p < ), and the increase of concentrations and exposure time increased mortality. The amount of mortality at high concentrations was remarkable, and highest mortality rate (96.91%) was recorded at 10 ml/kg, after 72 h of exposure. The application of oil significantly reduced F 1 progeny production and even at lowest concentration (2 ml/kg), the amount of F 1 production was reduced more than 70%, compared with control treatment. Complete (100%) reduction in progeny production was recorded at the rates of 8 and 10 ml/kg. The results of seed germination assay showed no significant differences between control and treated seed, and no harmful effect was observed on the seed germination. These results proved that Poppy seed oil can be used as a controlling agent of storage pests, especially C. maculatus, although more detailed studies are necessary. Keywords: Poppy seed oil; Callosobruchus maculatus; progeny production Introduction Pulses (family Leguminosae) are made up of many species, which are cultivated all over the world (Ofuya & Akhidue 2005). These crops play a major role in the daily diet of low-income groups of people (Rohitha Prasantha et al. 2003). Cowpea, Vigna unguiculata (L.) Walp, is one of the five most important legumes in the tropics, and provides protein for most people and nitrogen to the soils (Umeozor 2005). The production of cowpea is restricted by a number of biotic and abiotic factors, both in the field and the seed in storage. Among the constraining biotic factors are insect pests (Swella & Mushobozy 2007). The cowpea bruchid, Callosobruchus maculatus (F.) (Coleoptera: Bruchidae), is a cosmopolitan field-to-store pest, ranked as the principal post-harvest *Corresponding author. adel.khashaveh@gmail.com Ó 2013 Taylor & Francis

4 2 H. Sakenin Chelav and A. Khashaveh pest of cowpea. It causes substantial quantitative and qualitative losses, manifested by seed perforation, and reductions in weight, market value and germinability of seeds. Under traditional storage conditions, 100% infestation of cowpea, occurring within 3 5 months of storage, is common (Lale & Mustapha 2000). Residual insecticides have been used routinely for many years to control insect pests in stored grain. These insecticides are primarily organophosphorous and pyrethroid compounds (Arthur 1996). Several methods have been used over the years to protect cowpea grains from pest damages, especially C. maculatus, in storage; but, the use of residual insecticides has been very dominant (Abdullahi et al. 2011). However, use of residual insecticides is becoming less desirable because of the resistance in major insects, regulatory restrictions on use of insecticides, awareness of environmental pollution, the increasing of insecticides cost, erratic supplies, worker safety and consumer desire for a pesticide-free product, which has led to pest management specialists reappraising natural products (Arthur 1996; Haghtalab et al. 2009). The use of plant oils in the control of storage insect pests is an ancient practice and is particularly appropriate in small-scale storage systems. In recent years, increasing attention has been given to control of storage pests in grain depots by the use of oils, including vegetable oils, essential oils and mineral oils (Obeng-Ofori 1995; Obeng- Ofori & Reichmuth 1999; Nikpay 2007; Boateng & Kusi 2008; Haghtalab et al. 2009; Tabu et al. 2012; Lorestani et al. 2013). Poppy seed oil is an edible oil from opium Poppy (Papaver somniferum L.: Papaveraceae) seeds. The oil has culinary and pharmaceutical uses, as well as long established uses in the making of paints, varnishes and soaps. Poppy seeds yield 45 50% oil; it is highly palatable, high in vitamin E and has no narcotic properties (Wikipedia 2013). The object of our study was to evaluate the efficacy of Poppy seed oil as a natural insecticide against adults of C. maculatus in cowpea. According to our best knowledge, this is the first report about the use of plant materials from family Papaveraceae against insect pests. Materials and methods Insect rearing The initial stock culture of C. maculatus was obtained from Iranian Institute of Plant Protection that was maintained for at least two years, with no history of exposure to insecticides. C. maculatus was reared on cowpea seeds. Cowpea grains were disinfested by storing at 24 C for 72 h. Fifty pairs of C. maculatus adults were reared in 2-L wide-mouth glass jars containing 300 g of cowpea seeds. Insect cultures were incubated at C and 65 ± 5% relative humidity (r.h.) in continuous darkness. All adults used in the experiments were 0 24 h old and of mixed sex. For getting 0 24 h old adults, seeds with pupa window were separated and after one day, adults that emerged were collected with a hand-made aspirator. Poppy seed oil Poppy seed oil (extracted by cold-pressing) was purchased from Eghlyma Company, Tehran, Iran.

5 Archives of Phytopathology and Plant Protection 3 Adults mortality Poppy seed oil was applied at dose rates of 2, 4, 6, 8 and 10 ml/kg. Five lots of 1 kg of cowpea seed were prepared and placed in separate cylindrical jars (3-L capacity with screwed lids) and treated with appropriate dose rates, and one untreated lot served as a control. Subsequently, the jars were shaken manually for approximately 2 min to obtain an even distribution of oil on the entire seed sample. After a day, four samples of 50 g each, were taken from each jar (lot) as replication and then placed in glass vials (10.5 cm height and 6.5 cm diameter). After this, 25 adults were introduced into each glass vial that was covered with muslin cloth to provide sufficient aeration. The vials were then placed in incubators, set at C and 65 ± 5% r.h., in darkness. The number of dead adults was counted after 24, 48 and 72 h of exposure. Insects were considered dead when no response of leg or antennal movements to hot needle was observed. Progeny reduction percentage After the mortality count, all adults (dead and alive) were removed from the vials and the vials were left in the incubators at the same conditions for further days to assess progeny production. The number of emerged adults was then counted in controls and treated vials, and they were introduced to the below formula for the calculation of percentage of reduction in progeny production (Aldryhim 1990): Progeny reduction % ¼ No. progeny in control No. progeny in treatment 100 No. progeny in control Seed germination For determination of probable effect of Poppy seed oil on germination rate of cowpea seed, an assay was done with mentioned concentrations. For this purpose, 50 g of cowpea seed was treated with appropriate dose rates and kept in incubator at C and 65 ± 5% r.h. After one day, and for each dose rate, 50 seeds were taken and placed in 5 Petri dishes (10 seeds per each Petri dish) with moist filter paper as replications. The Petri dishes were kept in incubator for seven days, after which the number of germinated seeds was recorded. Statistical analysis The experiments were carried out, based on factorial experiment by randomized complete design with four replications. Mortality counts were corrected by using Abbott s formula (Abbott 1925). To equalise variances, mortality percentage p ffiffi of adults and number of progeny production were transformed using arcsine x. In contrast, the germination percentage of cowpea seed was not transformed. The data were analysed using analysis of variance (SAS 2000), with insect mortality as the response variable and dose rate and exposure time as main effects. The same procedure was carried out for progeny production counts. The means were separated by Duncan s multiple range test p = The concentration required to kill 50% of the insects (LC50) was estimated using probit analysis (SPSS 1999).

6 4 H. Sakenin Chelav and A. Khashaveh Table 1. ANOVA parameters for main effects and associated interaction for adults mortality in cowpea treated with Poppy seed oil. Source df F value P Dose rate < Exposure time < Dose rate Exposure time Error 45 Corrected total 59 Results Adults mortality For adults mortality, main effects were significant at the p<0.01 level, but associated interaction (dose rate exposure time) was not significant (Table 1). Mortality percentage increased with increase of dose rates and exposure time, which is quite rational. Lowest and highest mortality were observed after 24 h of exposure and 2 ml/kg and 72 h of exposure and 10 ml/kg of Poppy seed oil, respectively. These amounts were recorded as 4.12 ± 1.97 and ± 1.97%, respectively (Figure 1). The linear relationship between the exposure times and adults mortality was attained using probit analysis. The test for the goodness of fit indicates no significant heterogeneity in the linear relationships, for all the exposure times tested (small χ 2 values, p > 0.05). Based on the estimates of LC 50, and associated 95% confidence limits and no observed overlap, there was significant difference between the three exposure times (Table 2). The lowest amount of LC 50 value was recorded as 4.53 ml/kg after 72 h exposure. Progeny reduction percentage For progeny reduction, significant difference was noted between dose rates at the p <0.01 level, whereas exposure time had no significant effect on progeny reduction percentage. Also, associated interaction (dose rate exposure time) was not significant Figure 1. Mean mortality (% + SE) of C. maculatus adult treated with Poppy seed oil after 24, 48 and 72 h of exposure. Note: Means followed by the same letter are not significantly different (Duncan s Multiple Range Test at p = 0.05).

7 Archives of Phytopathology and Plant Protection 5 Table 2. The lethal concentration for the 50% (LC 50 ) in cowpea treated with Poppy seed oil. Time (h) LC 50 (ml/kg) 95% confidence Limits LC 95 (ml/kg) 95% confidence limits χ 2 P Slop (b) Intercept (a) ( ) ( ) ( ) ( ) ( ) ( ) Table 3. ANOVA parameters for main effects and associated interaction for progeny reduction percentage in Cowpea treated with Poppy seed oil. Source df F value P Dose rate < Exposure time Dose rate Exposure time Error 45 Corrected total 59 Figure 2. Mean percentage of reduction (% + SE) in progeny production (F 1 )ofc. maculatus treated with Poppy seed oil after 24, 48 and 72 h of exposure. Note: Means followed by the same letter are not significantly different (Duncan s Multiple Range Test at p = 0.05). (Table 3). The mean number of emerged adults (±SE) per vial in the control group was ± The application of oil significantly reduced F 1 progeny production, and even at lowest concentration (2 ml/kg) the amount of emerged adults was reduced more Table 4. Effect of Poppy seed oil on germination percentage of cowpea at different dose rate. Dose rate (ml/kg) Seed germination ±(SE) % ± 0.00 a ± 2.5 a ± 2.5 a ± 2.9 a ± 4.8 a Control group 97.5 ± 2.5 a Note: Means followed by the same letter are not significantly different (Duncan's Multiple Range Test at p = 0.05).

8 6 H. Sakenin Chelav and A. Khashaveh than 70%, compared with control treatment. Complete (100%) suppression in progeny production was recorded at the rates of 8 and 10 ml/kg (Figure 2). Seed germination The results of seed germination assay demonstrated no significant differences between control and treated seed, and no harmful effect was observed on the seed germination (Table 4). Discussion In this study, vegetable oils extracted from Poppy seed, were applied at different dose rates against cowpea bruchid. The main, and most important, issue in the management of storage pests is the use of safe and low-cost materials and methods, as most of the damage to stored products occurs in poor and developing countries. As mentioned in the introduction, Poppy seed oil is an edible oil that is obtained from opium poppy. Poppy is cultivated mainly for the legal pharmaceutical use of opium latex in India, USA, Egypt, Poland, Germany, Netherlands, China, Japan, Iran, Argentina, Spain, Bulgaria, Hungary and Portugal (Peter 2001). In these countries, the use of Poppy seed and its products are of secondary importance. In some cases, seeds are not used. So, extraction of oil from Poppy seed, as a secondary product of poppy culture, can be used to control pests. On the other hand, this is an edible oil that can be extracted by low-cost methods, such as cold-pressing, and had no harmful effects on human. In our study, Poppy seed oil has created considerable mortality in adults of cowpea bruchid, and highest mortality was achieved at the dose rate of 10 ml/kg, after 72 h of exposure. Reducing insect population in the next generation is an important aspect for the management of storage pest. The percentage of reduction in progeny production of cowpea bruchid in Poppy seed oil-treated cowpea, in comparison with untreated ones, was remarkable, and complete suppression of progeny production (F 1 ) was achieved at two high dose rates (8 and 10 ml/kg). At low doses (2 and 4 ml/kg), a large number of eggs were observed in the oil-treated cowpea, but very few eggs were hatched and emerged as F 1 progeny. On the other hand, a few eggs were found in the seeds that were treated with high doses (8 and 10 ml/kg), but none of them were hatched. Fewer eggs can be attributed to the high mortality of parents. Therefore, from these results, it can be concluded that the oil is prevented from hatching of eggs and finally, the number of offspring is reduced in the next generation. The mode of action of oils is yet to be confirmed, but most appear to cause death in the insect egg, larvae or adult by suffocation. The action of vegetable oils is by suffocation, but in other cases, oils can act as antifeedants, or even as insect growth regulators, by affecting metamorphosis (Don-Pedro 1989; Weaver & Subramanyam 2000). Oils caused high mortality of eggs and larvae on the seed surface, but had no effect on individual larvae that successfully entered the seed (Swella & Mushobozy 2007). Several published dates are available on the insecticidal effect of vegetable oils against stored grain insect pests, especially C. maculatus (Pacheco et al. 1995; Adedire & Ajayi 2003; Bamaiyi et al. 2006; Law-Ogbomo & Egharevba 2006; Haghtalab et al. 2009; Ilesanmi & Gungula 2010; Tabu et al. 2012). However, there are not any references related to plant materials, belonging to family Papaveraceae, to be compared with the results obtained in the present study; but since most of plant oils have same mode of action, they can be compared with each other. Pacheco et al. (1995) applied refined

9 Archives of Phytopathology and Plant Protection 7 soybean and crude castor oils at the rate of 0, 5 and 10 ml/kg of seeds, for the control of C. maculatus and C. phaseoli in stored chick-pea, and reported that both oils inhibited population growth of the two insect species, as compared to untreated seeds. They also noted that these oils had no harmful effect on the germination of chick-pea seeds. In another study, carried out by Haghtalab et al. (2009), the effect of different concentrations of castor and hazelnut oil were tested on C. maculatus in stored cowpea for evaluation of adults mortality and progeny production. These researchers reported that adults mortality received to 80.83% on hazelnut oil at the rate of 9.2 ml/kg and 86.66% on castor oil at the rate of 9 ml/kg, after 72 h of exposure. Also, in all treatment, oils reduced up to 90% of progeny production, in comparison with untreated cowpea. Law-Ogbomo and Egharevba (2006) evaluated vegetable oils from rubber seed oil, palm oil and palm kernel oil at the rates of 0, 2.5, 5, 7.5 and 10 ml/kg of cowpea seeds against C. maculatus in three cowpea varieties. The results obtained by these investigators indicated that all the plant oils tested have toxic effect on adults of C. maculatus in all cowpea varieties, and high mortality (72 100%) was achieved after 10 days of treatment. Khaya senegalensis (Desr.) seed oil was evaluated for control of C. maculatus on stored cowpea by Bamaiyi et al. (2006). They compared this plant oil with Pirimiphos methyl (E.C.) as a standard. They recorded complete adult mortality of C. maculatus within 24 h after treatment with seed oil at 1, 2 and 3 ml/100 g of cowpea. The seed oil significantly reduced the emergence of F 1 and F 2 progeny. They also demonstrated that for all parameters examined, no significant difference was observed between K. senegalensis seed oil and Pirimiphos methyl, in the control of C. maculatus. All of these finding confirm our results and indicate that vegetable oils are highly effective in the control of C. maculatus. Results obtained by Adedire and Ajayi (2003) show that Hura crepitans L. seed oil significantly reduced oviposition and adult emergence of C. maculatus. They recorded 100% ovicidal activity at all concentrations (0.02, 0.1, 0.2, 0.3 and 0.4 ml per 20 g of cowpea seed), while 100% larvicidal effect was observed in the last three concentrations. In our experiments, we observed that the concentration of 10 ml/kg is an appropriate rate that provides uniform coverage, while higher concentrations soaked the beans thoroughly. This may cause problems during storage and marketing. So, we need to use oils that have higher performance in lower concentrations. In conclusion, according to the results obtained in this experiment, Poppy seed oil can be recommended for control of C. maculatus, although further experiments are needed to assess the persistence and negative impacts of Poppy seed oil on storage materials. Acknowledgements This research was fully funded by the Young Researchers Club, Qaemshahr Branch, Islamic Azad University, Iran (Project Serial Number: 91194). The authors thank Dr Hamid Salehian (the head of Young Researchers Club, Qaemshahr Branch) for complete support of this research work. The resources made available by Islamic Azad University, Qaemshahr Branch, are also gratefully acknowledged. References Abbott W A method of computing the effectiveness of an insecticide. J Econ Entomol. 18: Abdullahi N, Majeed Q, Oyeyi TI Studies on the efficacy of Vittallaria paradoxa seed oil on the oviposition, hatchability of eggs and emergence of Callosobruchus maculatus (F.) (Coleoptera: Bruchidae) on treated cowpea seed. J Entomol. 8:

10 8 H. Sakenin Chelav and A. Khashaveh Adedire CO, Ajayi OE Potential of sandbox, Hura crepitans L. seed oil for protection of cowpea seeds from Callosobruchus maculatus Fabricius (Coleoptera: Bruchidae) infestation. J Plant Dis Prot. 110: Aldryhim Y Efficacy of the amorphous silica dust, Dryacide, against Tribolium confusum Duv. and Sitophilus granarius (L.) (Coleoptera: Tenebrionidae and Curculionidae). J Stored Prod Res. 26: Arthur F Grain protectants: current status and prospects for the future. J Stored Prod Res. 32: Bamaiyi LJ, Ndams IS, Toro WA, Odekina S Effect of mahogany Khaya senegalensis seed oil in the control of Callosobruchus maculatus on stored cowpea. Plant Prot Sci. 42: Boateng SA, Kusi F Toxicity of Jatropha seed oil to Callosobruchus maculatus (Coleoptera: Bruchidae) and its Parasitoid, Dinarmus basalis (Hymenoptera: Pteromalidae). J Appl Sci Res. 4: Don-Pedro KN Mechanism of action of some vegetable oils against Sitophilus zeamais Motsch. (Coleoptera: Curculionidae) on wheat. J Stored Prod Res. 25: Haghtalab N, Shayesteh N, Aramideh S Insecticidal efficacy of Castor and Hazelnut oils in stored Cowpea against Callosobruchus maculatus (F.) (Coleoptera: Bruchidae). J Biol Sci. 9: Ilesanmi JO, Gungula DT Preservation of Cowpea (Vigna unguiculata (L.) Walp.) grains against Cowpea Bruchids (Callosobruchus maculatus) using neem and moringa seed oils. Int J Agron. Article ID , 8 pages. Lale NES, Mustapha A Potential of combining neem (Azadirachta indica A. Juss.) seed oil with varietals resistance for the management of the cowpea bruchid, Callosobruchus maculatus (F.). J Stored Prod Res. 36: Law-Ogbomo KE, Egharevba RKA The use of vegetable oils in the control of Callosobruchus maculatus (F.) (Coleoptera: Bruchidae) in three cowpea varieties. Asian J Plant Sci. 5: Lorestani FA, Khashaveh A, Lorestani RA Fumigant toxicity of essential oil from Tanacetum balsamita L. (Compositae) against adults and eggs of Callosobruchus maculatus F. (Coleoptera: Bruchidae). Arch. Phytopathol. Plant Prot. (Accepted manuscript). Nikpay A Insecticidal efficacy of three vegetable oils as post-harvest grain protectants of stored wheat against Rhizopertha dominica (F.) (Coleoptera: Bostrychidae). Insect Sci. 14: Obeng-Ofori D Plant oils as grain protectants against infestations of Cryptolestes pusillus and Rhizopertha dominica in stored grain. Entomol Exp Appl. 77: Obeng-Ofori D, Reichmuth CH Plant oils as potentiation agents of monoterpenes for protection of stored grains against damage by stored product beetle pests. Int J Pest Manage. 45: Ofuya Z, Akhidue V The role of pulses in human nutrition: A review. J Appl Sci Environ Mgt. 9: Pacheco LA, De Castro MFP, De Paula DC, Lourencao AL, Bolonhezi S, Barbieri MK Efficacy of soybean and castor oils in control of Callosobruchus maculatus (F.) and Callosobruchus phaseoli (Gyllenhal) in stored chickpeas (Cicer arietinum L.). J Stored Prod Res. 31: Peter KV Handbook of herbs and spices. Cambridge: Woodhead. Rohitha Prasantha B, Reichmuth C, Buttner C Effect of temperature and relative humidity on diatomaceous earth treated Callosobruchus maculatus (F.) and Acanthoscelides obtectus (Say) (Coleoptera: Bruchidae). In: Proceedings of 8th International Working Conference on Stored product Protection; 2002 Jul 22 26; York, England. Wallingford, Oxon: CAB International; p. 1075, SAS Institute The SAS System version 8 for Windows, Cary, NC, USA. SPSS SPSS for Windows User s Guide Release 10. Chicago (IL): SPSS Inc. Swella GB, Mushobozy DMK Evaluation of the efficacy of protectants against Cowpea bruchids (Callosobruchus maculatus (F.)) on Cowpea seeds (Vigna unguiculata (L.) Walp.). Plant Prot Sci. 43:68 72.

11 Archives of Phytopathology and Plant Protection 9 Tabu D, Selvaraj T, Singh SK, Mulugeta N Management of Adzuki bean beetle (Callosobruchus chinensis L.) using some botanicals, inert materials and edible oils in stored chickpea. J Agric Technol. 8: Umeozor OC Effect of the infection of Callosobruchus maculatus (Fab.) on the weight loss of stored Cowpea (Vigna unguiculata (L.) Walp.). J Appl Sci Environ Manage. 9: Weaver DK, Subramanyam B Botanicals. In: Subramanyam BH, Hagstrum DW, editors. Alternatives to pesticides in stored-product IPM. Dordrecht: Kluwer Academic; p Wikipedia Poppy seed oil. Available from: