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Toxicity and repellency of Hoslundiaopposita Vahl (Lamiaceae) leaves' essential oil against rust-red flour beetle, Triboliumcastaneum Herbst (Coleoptera: Tenebrionidae) a
b
Samuel Adelani Babarinde , Adeyemi Oluseye Akinyemi , c
a
Lamidi Ajao Usman , Adeola Foluke Odewole , Abraham Opeola d
a
Sangodele , Oluwaseun Olasupo Iyiola & Oluwatoyin Deborah a
Olalere a
Department of Crop and Environmental Protection, Ladoke Akintola University of Technology, P.M.B. 4000, Ogbomoso, Nigeria b
Department of Agronomy, College of Agriculture, Osun State University, Ejigbo Campus, P.M.B. 4494, Osogbo, Nigeria c
Department of Chemistry, University of Ilorin, P.M.B. 1515, Ilorin, Nigeria d
Department of Crop Production and Soil Science, Ladoke Akintola University of Technology, P.M.B. 4000, Ogbomoso, Nigeria Published online: 14 Jan 2014.
To cite this article: Samuel Adelani Babarinde, Adeyemi Oluseye Akinyemi, Lamidi Ajao Usman, Adeola Foluke Odewole, Abraham Opeola Sangodele, Oluwaseun Olasupo Iyiola & Oluwatoyin Deborah Olalere (2014) Toxicity and repellency of Hoslundiaopposita Vahl (Lamiaceae) leaves' essential oil against rust-red flour beetle, Triboliumcastaneum Herbst (Coleoptera: Tenebrionidae), Natural Product Research: Formerly Natural Product Letters, 28:6, 365-371, DOI: 10.1080/14786419.2013.866115 To link to this article: http://dx.doi.org/10.1080/14786419.2013.866115
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Natural Product Research, 2014 Vol. 28, No. 6, 365–371, http://dx.doi.org/10.1080/14786419.2013.866115
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Toxicity and repellency of Hoslundia opposita Vahl (Lamiaceae) leaves’ essential oil against rust-red flour beetle, Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) Samuel Adelani Babarindea*, Adeyemi Oluseye Akinyemib, Lamidi Ajao Usmanc, Adeola Foluke Odewolea, Abraham Opeola Sangodeled, Oluwaseun Olasupo Iyiolaa and Oluwatoyin Deborah Olalerea a Department of Crop and Environmental Protection, Ladoke Akintola University of Technology, P.M.B. 4000, Ogbomoso, Nigeria; bDepartment of Agronomy, College of Agriculture, Osun State University, Ejigbo Campus, P.M.B. 4494, Osogbo, Nigeria; cDepartment of Chemistry, University of Ilorin, P.M.B. 1515, Ilorin, Nigeria; dDepartment of Crop Production and Soil Science, Ladoke Akintola University of Technology, P.M.B. 4000, Ogbomoso, Nigeria
(Received 20 September 2013; final version received 4 November 2013) The insecticidal properties of Hoslundia opposita Vahl (Lamiaceae) leaves’ essential oil (EO) against Tribolium castaneum were investigated using contact toxicity and repellency bioassays. Mortality progressed with exposure period and ranged from 61.13% observed at 24 h after treatment (HAT) to 88.86% at 168 HAT. The LT50 (lethal time for 50% of treated adults) of H. opposita EO against T. castaneum was 10.42 h. Application of EO at 20 – 30 mL/30 cm2 caused significantly (P , 0.05) higher percentage repellency than what was observed in control, 10 and 15 mL/30 cm2 at 1 and 3 HAT, with significant repellency at 24 HAT regardless of dosage. Repellency class increased with EO dosage, with class V observed at 30 mL/30 cm2, regardless of exposure duration. The RD50 (repellency dose for 50% of treated adults) of 15.88 and 13.37 mL/30 cm2 for 1 and 2 HAT, respectively, was significantly higher than 0.09 mL/30 cm2 at 24 HAT. Keywords: Rust-red flour beetle; insecticidal; repellency; essential oil; Hoslundia opposita
1. Introduction Tribolium castaneum Herbst, commonly called rust-red flour beetle, is an important world-wide pest of stored products that is observed among several commodities. It may cause considerable economic losses if not adequately controlled because it has a very high rate of population increase (Hill 1990) and can also attack grains embryo. It is particularly troublesome on cereal flour on which it imparts a brownish tinge and a pungent smell due to the secretion of benzoquinones by females from a pair of abdominal defence gland (Haines 1991). Pearl millet [Pennisetum glaucum (L.) R. Br.] belongs to Family Poaceae and is today the world’s sixth most important cereal grain. It is a traditionally major food crop grown in arid and semi-arid tropical environment of Indian subcontinent and Africa (Rai et al. 2007). The crop is a major source of energy (carbohydrate), protein, vitamin and minerals for millions of poor people in the region. It has high-quality nutrients for humans and livestock and is considered more efficient on utilisation of soil moisture than sorghum and maize. Despite its many uses, both the larvae and
*Corresponding author. Emails: [email protected]; [email protected] q 2014 Taylor & Francis
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the adult of the rust-red flour beetle (T. castaneum) have been reported to attack it (Turaki et al. 2007; Babarinde & Ogunkeyede 2008). Due to the intensive infestation of T. castaneum in millet and other stored commodities, there is need for the control of the pest to alleviate the food security problem in the third world. This has led to the continued search for humanly safe, host-specific, cost-effective and ecologically tolerable means of managing the pest. The acceptance of botanical insecticides for the control of storage insect pest by small-scale farmers is influenced by the following parameters: availability, safety, quality and cost (Adedire 2003). Hoslundia opposita Vahl (Lamiaceae) is a herbaceous perennial shrub widely distributed in Africa (Morton 1963) where its various parts are being used in traditional medicine for the treatment of herpes, conjunctivitis, epilepsy, chest pain, yellow fever and stomach troubles (Ayensu 1978; Gelfand et al. 1985). Infusions of its leaves have been used as a purgative, diuretic, febrifuge, antibiotic and antiseptic (Mujovo et al. 2008). Non-volatile compounds identified from its leaves were mainly flavonoids (Ngadjui et al. 1995). Its other uses include antibacterial activity (Ngadjui et al. 1995). Early work by Usman et al. (2010) revealed that the essential oil (EO) of H. opposita leaves is monoterpene-dominated, with 1,8-cineole being the predominant constituent. These terpenoids have been reported to be insecticidal against many stored product pests (Obeng-Ofori et al. 1997). Mujovo et al. (2008) reported that H. opposita inhibited the HIV-1 reverse transcriptase enzyme. Despite the various reported uses of H. opposita, there is a need to investigate and establish its new potentials. Therefore, the research was designed to investigate the insecticidal properties of H. opposita against T. castaneum, a cosmopolitan post-harvest pest of pearl millet. 2. Results and discussion 2.1. Contact toxicity of H. opposita EO against adult T. castaneum Several authors (Jibilou et al. 2008; Mondal & Khalequzzaman 2009; Babarinde et al. 2011) have reported the use of insecticidal plant products as protectants against T. castaneum, being aspect of integrated plant protection within the frame work of ecologically sound and economically reasonable schemes. Figure 1 shows that T. castaneum was susceptible to contact application of H. opposita. When insects were exposed for 24 h, 61.13% mortality was recorded which was significantly higher than 0% mortality observed in the control. Mortality progressed with exposure period with the highest percent mortality (88.86%) observed when insects were exposed for 168 h. The LT50 of the EO of H. opposita against adult T. castaneum was 10.42 h with 0.49 and 26.20 as lower and upper fiducial limits respectively. The result indicates that the contact toxicity of EO of H. opposita against T. castaneum adults was exposure period-dependent. This observation conforms to the findings of Babarinde et al. (2008) who reported that the toxicity of Tithonia diversifolia against Sitophilus zeamais was exposure period-dependent. Stamopolous et al. (2007) reported that the toxicity of 1,8cineole to 10-day-old adult male and female T. confusum was dose- and exposure period-dependent. The exposure period-dependence toxicity observed in this study is also in conformity with Athanassiou et al. (2012) who reported the mortality of S. zeamais and T. confusum exposed to the mixture of silica gel and Juniperus oxycedrus ssp. oxycedrus EO. The reason for the exposure period-dependent toxicity was that the EO penetrated the cuticle of the tergum and get into the body system to affect acetylcholinesterase. Terpenoids including 1,8-cineole showed a reversible competitive inhibition of acetylcholinesterase by apparently occupying the hydrophobic site of the enzyme’s active centre (Obeng-Ofori et al. 1998). 2.2. Effect of H. opposita on repellency of T. castaneum It was observed that the longer the T. castaneum was exposed to H. opposita oil, the higher they were repelled. Application of EO at 20 – 30 mL/30 cm2 caused significantly higher percentage
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96
72
48
24 0
10
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30
40
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Figure 1. Contact toxicity of EO of H. opposita against adult T. castaneum (EO applied at 0.1 mL per beetle, 10 beetles per replicate, 3 replicates for the bioassay).
repellency (PR) than what was observed in control, 10 and 15 mL/30 cm2 at 1 h (df ¼ 5, 18, F ¼ 6.561, P ¼ 0.001) and 3 h (df ¼ 5, 18, F ¼ 9.799, P , 0.001) after exposure, while at 24 h after exposure, application of EO regardless of the dose causes significant (df ¼ 5, 18, F ¼ 54.341, P , 0.000) repellency compared with the control (Table 1). Repellency class increased with EO dosage, with repellency class V observed at the application of 30 mL/30 cm2, regardless of the exposure duration. The RD50 decreased with exposure duration. It was 15.88 (14.70 – 17.67) and 13.37 (11.59 –14.97) for 1 and 3 h after treatment (HAT), respectively, which was significantly higher than 0.09 h (0.06 –1.00) obtained at 24 HAT (there was no overlap in the fiducial limit) (Table 1). The result of the repellency bioassay indicates that the response of the insect was dependent on the dosage and time of exposure. This is because the bioactive components of H. opposita did not volatilise because the repellency chamber was closed. Also, there was hyper excitability due to the olfaction of the EO at the early exposure period, but the insects attained stability in their responses to the impact of the EO with an increase in exposure duration. Hence, higher PR was observed at a later exposure period than early exposure period. This implies that the EO did not evaporate immediately after application; therefore, it can be absorbed by millet seeds and repel the beetle with persistence when applied for pest control. The repellent bioactivity implies that the EO can prevent the non-resident T. castaneum population. Babarinde and Adeyemo (2010) had earlier reported the repellency of Xylopia aethiopica extract against T. castaneum. Similarly, AbdelSattar et al. (2010) reported the insecticidal and repellent activity of Schinus molle L. on Trogoderma granarium and T. castaneum. The fact that the EO repelled and killed the insects elucidates its potentials for the control of T. castaneum in stored millet seed. The Nigerian grown H. opposita had been reported to be 1,8-cineole chemotype (Usman et al. 2010). The insecticidal properties of the EO were due to the presence of its chemical constituents, with 1,8-cineole being the predominant. Several authors (Obeng-Ofori
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Table 1. PR of adult T. castaneum exposed to H. opposita EO and RD50 at different exposure period. Duration (h)
Treatment (mL/30 cm2)
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Control (0) 10 15 20 25 30 ANOVA result RD50a
1
2
24
0.00a (0) 26.00ab (II) 45.00abc (III) 60.85bc (IV) 67.50bc (IV) 83.35c (V) df ¼ 5, 18, F ¼ 6.561, P ¼ 0.001 15.88 (14.70 –17.67)
0.00a (0) 35.20ab (II) 60.85bc (IV) 66.90bc (IV 74.15bc (IV) 90.00c (V) df ¼ 5, 18, F ¼ 9.799, P , 0.001 13.37 (11.59– 14.97)
0.00a (0) 80.20b (V) 90.00b (V) 83.38b (V) 90.00b (V) 90.00b (V) df ¼ 5, 18, F ¼ 54.341, P , 0.001 0.09 (0.062 –1.00)
Note: Data are means of four replicate. Means carrying the same letter along the column are not significantly different at 5% level of probability using Turkey H.S.D. Roman figures in parentheses are repellency classes. Repellency class 0: ,0.1%; Repellency class I: 0.1–20%. Repellency class II: 20.1–40%; Repellency class III: 40.1–60%. Repellency class IV: 60.1– 80%; Repellency class V: 80.1–100%. a For RD50, Arabic numerals in parentheses are lower and upper fiducial limits, respectively.
et al. 1997; Stamopolous et al. 2007; Mossi et al. 2011) have reported the insecticidal properties of botanicals rich in 1,8-cineole against insects causing damage to stored products. 3. Experimental 3.1. Experimental site The experiment was carried out at the Storage Entomology Unit of Crop and Environmental Protection Laboratory, Ladoke Akintola University of Technology (LAUTECH), Ogbomoso, Nigeria. 3.2. Collection and handling of millet seeds Pearl millet seeds harvested in January 2011 from Bida, Niger State, Nigeria, were obtained from Wazobia Market, Ogbomoso, Nigeria, in April 2011 and were disinfested by storage at 48C for 2 weeks before use. Thereafter, the millet seeds were placed on laboratory bench for 2 days to equilibrate with the ambient relative humidity before use for the experimental purpose. 3.3. Culturing of T. castaneum T. castaneum was obtained from a culture maintained in Storage Entomology Unit of Crop and Environmental Protection Laboratory, LAUTECH, Ogbomoso, Nigeria. Insect rearing was done in the laboratory under ambient temperature of 25– 358C and relative humidity of 68 –80%, according to the method of Babarinde and Ogunkeyede (2008), using a portion of the millet seeds obtained from Wazobia Market, Ogbomoso. 3.4. Collection and extraction of EO from H. opposita leaves The leaves of H. opposita used for this study were obtained from Akoda Village, via Oyo, Nigeria, in September 2011. Authentication of the plant was done in the Department of Pure and Applied Biology, LAUTECH, Ogbomoso, where voucher specimen was dropped and a voucher number, LH0 368, was allocated. The plant material was air dried under shade in the laboratory until crisp, and later milled into fine powder using a milling machine. Two hundred and fifty grams of the powder was pre-soaked in 1000 mL of distilled water for 24 h, after which EO was obtained via
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hydrodistillation for 3 h with the aid of Clevenger apparatus according to the method recommended by British Pharmacopoeia (1988). The oil yield was 0.54% v/w. The EO obtained was stored in an air-tight glass sample bottle and kept in refrigerator until use. 3.5. Contact toxicity test Ten T. castaneum adults (1- to 7-day old) were selected for this bioassay. The contact toxicity bioassay followed the earlier method described by Babarinde and Ewete (2008) with some modifications. H. opposita EO at the rate of 1.0 mL was applied on the tergum of each insect with the aid of Hamilton syringe (Model 705 N, Sigmaw, St. Louis, USA). After the application of EO, the 10 insects were placed in 9 cm-diameter Petri dish and covered. A control experiment which was a collection of 10 T. castaneum adults similar in all respects as those used for the EO treatment was included. They were placed in 9 cm-diameter Petri dish and covered, without any treatment. The experiment was replicated three times. Mortality data were taken at 1, 24, 48, 72, 96, 120, 148 and 168 HAT. Insects that did not respond to pin probe were adjudged dead. Where there was mortality in the control, mortality was corrected with Abbott’s formula (Abbott 1925). Percentage mortality (PM) was calculated using the formula: PM ¼
Number of dead insects £ 100: Total number of experimented insects
3.6. Repellency test The method used for testing the repellency of H. opposita EO extract against T. castaneum adults was based on the area-preferred test described by Babarinde and Adeyemo (2010) with the following modifications. EO was applied at 10, 15, 20, 25 and 30 mL per half disc of (30 cm2) Whatman No. 1 filter paper in 9 cm-diameter Petri dishes, which corresponded to 0.33, 0.50, 0.66, 0.83 and 1.0 mL/cm2. The other half was left untreated. Control with 0.2 mL ethanol was also set up, with the other half of the filter paper left untreated. After 15 min of treatment of filter papers, 10 T. castaneum adults were introduced at the centre of the repellency bioassay chamber. There were four replicates for each treatment. Data on the repellence of EO against T. castaneum were taken at 1, 3 and 24 HAT. PR was calculated using the formula: PR ¼
Nu 2 Nt £ 100; Nu þ Nt
where Nu is the number of insects on untreated filter paper disc and Nt is the number of insects on treated filter paper disc. 3.7. Experimental design and data analysis The experiment was set up in completely randomised design. Data were transformed using angular transformation and subsequently subjected to analysis of variance (ANOVA) using SAS Package (SAS 2000). Where there was significant treatment effect, Tukey’s HSD test at 5% probability level was used to separate the means. Probit analyses were done to determine LT50 and RD50 for contact toxicity and repellency bioassays, respectively. 4. Conclusion The bioactivity of H. opposita EO against T. castaneum reveals its potentials to control the insect. This discovery can provide leads for active biopesticides for the protection of pearl millets against T. castaneum.
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Acknowledgement The authors acknowledge Dr A. T. J. Ogunkunle of Department of Pure and Applied Biology, LAUTECH, Ogbomoso, Nigeria, for authentication of H. opposita.
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References Abbott WS. 1925. A method for computing the effectiveness of an insecticide. J Econ Entomol. 18:265–267. Abdel-Sattar E, Zaitoun AA, Farag MA, El Gayed SH, Harraz FMH. 2010. Chemical composition, insecticidal and insect repellent activity of Schinus molle L. leaf and fruit essential oils against Trogoderma granarium and Tribolium castaneum. Nat Prod Res. 24(3):226–235. Adedire CO. 2003. Use of nutmeg, Myristica fragrans (Houtt) powder and oil for the control of cowpea storage bruchid, Callosobruchus maculatus (Fabricius). J Plant Dis Prot. 109:193–199. Athanassiou CG, Kavallieratos NG, Evergetis E, Katsoula AM, Haroutounian SA. 2012. Insecticidal efficacy of silica gel with Juniperus oxycedrus ssp. oxycedrus (Pinales: Cupressaceae) essential oil against Sitophilus oryzae (Coleoptera: Curculionidae) and Tribolium confusum (Coleoptera: Tenebrionidae). J Econ Entomol. 106 (4):1902–1910. Ayensu ES. 1978. Medicinal plants of West Africa. Algonac, MI: Reference Publications, 282 p. Babarinde SA, Adeyemo YA. 2010. Toxic and repellent properties of Xylopia aethiopica (Dunal) A. Richard on Tribolium castaneum Herbst infesting stored millets, Pennisetum glaucum (L.) R. Br. Arch Phytopathol Plant Prot. 43(8):810–816. Babarinde SA, Ewete FK. 2008. Comparative bioactivity of three Khaya species (Meliaceae) on Callosobruchus maculatus Fabricus (Coleoptera: Bruchidae). J Entomol Res Soc. 10(1):27–35. Babarinde SA, Ogunkeyede AF. 2008. Fecundity suppression of two botanicals on rust red flour beetle, Tribolium castaneum infesting sorghum. J Ultra Scientist Phys Sci. 20(2):411–413. Babarinde SA, Olabode OS, Akanbi MO, Adeniran OA. 2008. Potential of Tithonia diversifolia with Pirimiphos-methyl in control of Sitophilus zeamais (Coleoptera: Curculionidae). Afr J Plant Sci Biotechnol. 2(2):77–80. Babarinde SA, Oyegoke OO, Adekunle AE. 2011. Larvicidal and insecticidal properties of Ricinus communis seed extracts obtained by different methods against Tribolium castaneum Herbst (Coleoptera: Tenebrionidae). Arch Phytopathol Plant Prot. 44(5):451–459. British Pharmacopoeia. 1988. British Pharmacopoeia. Vol. 2. London: Her Majesty’s Stationery Office (HMSO); p. 137– 138. Gelfand M, Mavis D, Drummond RB, Ndemera N. 1985. The traditional medical practictioners in Zaimbawe, the principles, practice and pharmacopoeia. Gweru: Mambo Press. Haines CP. 1991. Insects and arachnids of tropical stored product their biology and identification. 2nd ed. Chatham, Kent: Natural Resources Institute. Hill DS. 1990. Pest of products and their control. London: Belhaven Press. Jibilou R, Amri H, Bouayad N, Ghailani N, Ennabili A, Sayah F. 2008. Insecticidal effects of extracts of seven plant species on larval development, a-amylase activity and offspring production of Tribolium castanuem (Herbst) (Insecta: Coleoptera: Tenebrionidae). Bioresour Technol. 99:959– 964. Mondal M, Khalequzzaman M. 2009. Ovicidal activity of essential oils against red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae). J Biosci. 17:57–62. Morton JK. 1963. In: Hutchinson J, Dalziel JM, editors. Flora of West Tropical Africa. 2nd ed. London, 450 p. Crown Agents. Mossi AJ, Astolfi V, Kubiak G, Lerin L, Zanella C, Toniazzo G, de Oliveira D, Treichel H, Devilla IA, Cansian R, Restello R. 2011. Insecticidal and repellency activity of essential oil of Eucalyptus species against Sitophilus zeamais Motschulsky (Coleoptera, Curculionidae). J Sci Food Agric. 91:273–277. Mujovo SF, Husseina AA, Marion Meyer JJ, Fourie B, Muthivh T, Lall N. 2008. Bioactive compounds from Lippia javanica and Hoslundia opposita. Nat Prod Res. 22(12):1047– 1054. Ngadjui BT, Tsopmo A, Ayafor JF, Connolly JD, Tamboue HH. 1995. New Pyrone substituted flavonoid from Hoslundia opposita. J Nat Prod. 58:109–111. Obeng-Ofori D, Reichmuth CH, Bekele J, Hassanali A. 1997. Biological activity of 1,8-cineole, a major component of essential oil of Ocimum kenyense (Ayobangira) against stored product beetles. J Appl Entomol. 121:237– 243. Obeng-Ofori D, Reichmuth CH, Bekele AJ, Hassanali A. 1998. Toxicity and protectant potential of camphor, a major component of essential oil of Ocimum kilmanscharicum, against four-stored product beetles. Int J Pest Manage. 44:203–209. Rai KN, Alur AS, Reddy CR, Kalapande HV, Birajdar SN, Reddy KMV, Reddy AR. 2007. Pearl millet production technologies for Andhra Pradesh and Maharastra. Global theme on crop improvement. Andhra Pradesh: International Crops Research Institute for the Semi-Arid Tropics, 16 p. SAS Institute. 2000. Statistical analytical systems SAS/STAT user’s guide version 8(2). Cary, NC: SAS Institute.
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Stamopolous DC, Damos D, Karagianidou G. 2007. Bioactivity of five monoterpenoid vapours to Triboluim confusum (du Val) (Coleoptera: Tenebrionidae). J Stored Prod Res. 43:571–577. Turaki JM, Sastawa BM, Kabir BGJ, Lale NES. 2007. Susceptibility of flours derived from various cereal grains to infestation by the rust-red flour beetle (Tribolium castaneum Herbst) (Coleoptera: Tenebrionidae) in different seasons. J Plant Prot Res. 47(3):279–288. Usman LA, Zubair MF, Adebayo SA, Oladosu IA, Muhammad NO, Akolade JO. 2010. Chemical composition of leaf and fruit essential oils of Hoslundia opposita Vahl grown in Nigeria. Am Eurasian J Agric Environ Sci. 8(1):40–43.
Natural Product Research: Formerly Natural Product Letters Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/gnpl20
Toxicity and repellency of Hoslundiaopposita Vahl (Lamiaceae) leaves' essential oil against rust-red flour beetle, Triboliumcastaneum Herbst (Coleoptera: Tenebrionidae) a
b
Samuel Adelani Babarinde , Adeyemi Oluseye Akinyemi , c
a
Lamidi Ajao Usman , Adeola Foluke Odewole , Abraham Opeola d
a
Sangodele , Oluwaseun Olasupo Iyiola & Oluwatoyin Deborah a
Olalere a
Department of Crop and Environmental Protection, Ladoke Akintola University of Technology, P.M.B. 4000, Ogbomoso, Nigeria b
Department of Agronomy, College of Agriculture, Osun State University, Ejigbo Campus, P.M.B. 4494, Osogbo, Nigeria c
Department of Chemistry, University of Ilorin, P.M.B. 1515, Ilorin, Nigeria d
Department of Crop Production and Soil Science, Ladoke Akintola University of Technology, P.M.B. 4000, Ogbomoso, Nigeria Published online: 14 Jan 2014.
To cite this article: Samuel Adelani Babarinde, Adeyemi Oluseye Akinyemi, Lamidi Ajao Usman, Adeola Foluke Odewole, Abraham Opeola Sangodele, Oluwaseun Olasupo Iyiola & Oluwatoyin Deborah Olalere (2014) Toxicity and repellency of Hoslundiaopposita Vahl (Lamiaceae) leaves' essential oil against rust-red flour beetle, Triboliumcastaneum Herbst (Coleoptera: Tenebrionidae), Natural Product Research: Formerly Natural Product Letters, 28:6, 365-371, DOI: 10.1080/14786419.2013.866115 To link to this article: http://dx.doi.org/10.1080/14786419.2013.866115
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to
the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content.
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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. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/termsand-conditions
Natural Product Research, 2014 Vol. 28, No. 6, 365–371, http://dx.doi.org/10.1080/14786419.2013.866115
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Toxicity and repellency of Hoslundia opposita Vahl (Lamiaceae) leaves’ essential oil against rust-red flour beetle, Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) Samuel Adelani Babarindea*, Adeyemi Oluseye Akinyemib, Lamidi Ajao Usmanc, Adeola Foluke Odewolea, Abraham Opeola Sangodeled, Oluwaseun Olasupo Iyiolaa and Oluwatoyin Deborah Olalerea a Department of Crop and Environmental Protection, Ladoke Akintola University of Technology, P.M.B. 4000, Ogbomoso, Nigeria; bDepartment of Agronomy, College of Agriculture, Osun State University, Ejigbo Campus, P.M.B. 4494, Osogbo, Nigeria; cDepartment of Chemistry, University of Ilorin, P.M.B. 1515, Ilorin, Nigeria; dDepartment of Crop Production and Soil Science, Ladoke Akintola University of Technology, P.M.B. 4000, Ogbomoso, Nigeria
(Received 20 September 2013; final version received 4 November 2013) The insecticidal properties of Hoslundia opposita Vahl (Lamiaceae) leaves’ essential oil (EO) against Tribolium castaneum were investigated using contact toxicity and repellency bioassays. Mortality progressed with exposure period and ranged from 61.13% observed at 24 h after treatment (HAT) to 88.86% at 168 HAT. The LT50 (lethal time for 50% of treated adults) of H. opposita EO against T. castaneum was 10.42 h. Application of EO at 20 – 30 mL/30 cm2 caused significantly (P , 0.05) higher percentage repellency than what was observed in control, 10 and 15 mL/30 cm2 at 1 and 3 HAT, with significant repellency at 24 HAT regardless of dosage. Repellency class increased with EO dosage, with class V observed at 30 mL/30 cm2, regardless of exposure duration. The RD50 (repellency dose for 50% of treated adults) of 15.88 and 13.37 mL/30 cm2 for 1 and 2 HAT, respectively, was significantly higher than 0.09 mL/30 cm2 at 24 HAT. Keywords: Rust-red flour beetle; insecticidal; repellency; essential oil; Hoslundia opposita
1. Introduction Tribolium castaneum Herbst, commonly called rust-red flour beetle, is an important world-wide pest of stored products that is observed among several commodities. It may cause considerable economic losses if not adequately controlled because it has a very high rate of population increase (Hill 1990) and can also attack grains embryo. It is particularly troublesome on cereal flour on which it imparts a brownish tinge and a pungent smell due to the secretion of benzoquinones by females from a pair of abdominal defence gland (Haines 1991). Pearl millet [Pennisetum glaucum (L.) R. Br.] belongs to Family Poaceae and is today the world’s sixth most important cereal grain. It is a traditionally major food crop grown in arid and semi-arid tropical environment of Indian subcontinent and Africa (Rai et al. 2007). The crop is a major source of energy (carbohydrate), protein, vitamin and minerals for millions of poor people in the region. It has high-quality nutrients for humans and livestock and is considered more efficient on utilisation of soil moisture than sorghum and maize. Despite its many uses, both the larvae and
*Corresponding author. Emails: [email protected]; [email protected] q 2014 Taylor & Francis
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the adult of the rust-red flour beetle (T. castaneum) have been reported to attack it (Turaki et al. 2007; Babarinde & Ogunkeyede 2008). Due to the intensive infestation of T. castaneum in millet and other stored commodities, there is need for the control of the pest to alleviate the food security problem in the third world. This has led to the continued search for humanly safe, host-specific, cost-effective and ecologically tolerable means of managing the pest. The acceptance of botanical insecticides for the control of storage insect pest by small-scale farmers is influenced by the following parameters: availability, safety, quality and cost (Adedire 2003). Hoslundia opposita Vahl (Lamiaceae) is a herbaceous perennial shrub widely distributed in Africa (Morton 1963) where its various parts are being used in traditional medicine for the treatment of herpes, conjunctivitis, epilepsy, chest pain, yellow fever and stomach troubles (Ayensu 1978; Gelfand et al. 1985). Infusions of its leaves have been used as a purgative, diuretic, febrifuge, antibiotic and antiseptic (Mujovo et al. 2008). Non-volatile compounds identified from its leaves were mainly flavonoids (Ngadjui et al. 1995). Its other uses include antibacterial activity (Ngadjui et al. 1995). Early work by Usman et al. (2010) revealed that the essential oil (EO) of H. opposita leaves is monoterpene-dominated, with 1,8-cineole being the predominant constituent. These terpenoids have been reported to be insecticidal against many stored product pests (Obeng-Ofori et al. 1997). Mujovo et al. (2008) reported that H. opposita inhibited the HIV-1 reverse transcriptase enzyme. Despite the various reported uses of H. opposita, there is a need to investigate and establish its new potentials. Therefore, the research was designed to investigate the insecticidal properties of H. opposita against T. castaneum, a cosmopolitan post-harvest pest of pearl millet. 2. Results and discussion 2.1. Contact toxicity of H. opposita EO against adult T. castaneum Several authors (Jibilou et al. 2008; Mondal & Khalequzzaman 2009; Babarinde et al. 2011) have reported the use of insecticidal plant products as protectants against T. castaneum, being aspect of integrated plant protection within the frame work of ecologically sound and economically reasonable schemes. Figure 1 shows that T. castaneum was susceptible to contact application of H. opposita. When insects were exposed for 24 h, 61.13% mortality was recorded which was significantly higher than 0% mortality observed in the control. Mortality progressed with exposure period with the highest percent mortality (88.86%) observed when insects were exposed for 168 h. The LT50 of the EO of H. opposita against adult T. castaneum was 10.42 h with 0.49 and 26.20 as lower and upper fiducial limits respectively. The result indicates that the contact toxicity of EO of H. opposita against T. castaneum adults was exposure period-dependent. This observation conforms to the findings of Babarinde et al. (2008) who reported that the toxicity of Tithonia diversifolia against Sitophilus zeamais was exposure period-dependent. Stamopolous et al. (2007) reported that the toxicity of 1,8cineole to 10-day-old adult male and female T. confusum was dose- and exposure period-dependent. The exposure period-dependence toxicity observed in this study is also in conformity with Athanassiou et al. (2012) who reported the mortality of S. zeamais and T. confusum exposed to the mixture of silica gel and Juniperus oxycedrus ssp. oxycedrus EO. The reason for the exposure period-dependent toxicity was that the EO penetrated the cuticle of the tergum and get into the body system to affect acetylcholinesterase. Terpenoids including 1,8-cineole showed a reversible competitive inhibition of acetylcholinesterase by apparently occupying the hydrophobic site of the enzyme’s active centre (Obeng-Ofori et al. 1998). 2.2. Effect of H. opposita on repellency of T. castaneum It was observed that the longer the T. castaneum was exposed to H. opposita oil, the higher they were repelled. Application of EO at 20 – 30 mL/30 cm2 caused significantly higher percentage
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120
96
72
48
24 0
10
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30
40
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% mortality
Figure 1. Contact toxicity of EO of H. opposita against adult T. castaneum (EO applied at 0.1 mL per beetle, 10 beetles per replicate, 3 replicates for the bioassay).
repellency (PR) than what was observed in control, 10 and 15 mL/30 cm2 at 1 h (df ¼ 5, 18, F ¼ 6.561, P ¼ 0.001) and 3 h (df ¼ 5, 18, F ¼ 9.799, P , 0.001) after exposure, while at 24 h after exposure, application of EO regardless of the dose causes significant (df ¼ 5, 18, F ¼ 54.341, P , 0.000) repellency compared with the control (Table 1). Repellency class increased with EO dosage, with repellency class V observed at the application of 30 mL/30 cm2, regardless of the exposure duration. The RD50 decreased with exposure duration. It was 15.88 (14.70 – 17.67) and 13.37 (11.59 –14.97) for 1 and 3 h after treatment (HAT), respectively, which was significantly higher than 0.09 h (0.06 –1.00) obtained at 24 HAT (there was no overlap in the fiducial limit) (Table 1). The result of the repellency bioassay indicates that the response of the insect was dependent on the dosage and time of exposure. This is because the bioactive components of H. opposita did not volatilise because the repellency chamber was closed. Also, there was hyper excitability due to the olfaction of the EO at the early exposure period, but the insects attained stability in their responses to the impact of the EO with an increase in exposure duration. Hence, higher PR was observed at a later exposure period than early exposure period. This implies that the EO did not evaporate immediately after application; therefore, it can be absorbed by millet seeds and repel the beetle with persistence when applied for pest control. The repellent bioactivity implies that the EO can prevent the non-resident T. castaneum population. Babarinde and Adeyemo (2010) had earlier reported the repellency of Xylopia aethiopica extract against T. castaneum. Similarly, AbdelSattar et al. (2010) reported the insecticidal and repellent activity of Schinus molle L. on Trogoderma granarium and T. castaneum. The fact that the EO repelled and killed the insects elucidates its potentials for the control of T. castaneum in stored millet seed. The Nigerian grown H. opposita had been reported to be 1,8-cineole chemotype (Usman et al. 2010). The insecticidal properties of the EO were due to the presence of its chemical constituents, with 1,8-cineole being the predominant. Several authors (Obeng-Ofori
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Table 1. PR of adult T. castaneum exposed to H. opposita EO and RD50 at different exposure period. Duration (h)
Treatment (mL/30 cm2)
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Control (0) 10 15 20 25 30 ANOVA result RD50a
1
2
24
0.00a (0) 26.00ab (II) 45.00abc (III) 60.85bc (IV) 67.50bc (IV) 83.35c (V) df ¼ 5, 18, F ¼ 6.561, P ¼ 0.001 15.88 (14.70 –17.67)
0.00a (0) 35.20ab (II) 60.85bc (IV) 66.90bc (IV 74.15bc (IV) 90.00c (V) df ¼ 5, 18, F ¼ 9.799, P , 0.001 13.37 (11.59– 14.97)
0.00a (0) 80.20b (V) 90.00b (V) 83.38b (V) 90.00b (V) 90.00b (V) df ¼ 5, 18, F ¼ 54.341, P , 0.001 0.09 (0.062 –1.00)
Note: Data are means of four replicate. Means carrying the same letter along the column are not significantly different at 5% level of probability using Turkey H.S.D. Roman figures in parentheses are repellency classes. Repellency class 0: ,0.1%; Repellency class I: 0.1–20%. Repellency class II: 20.1–40%; Repellency class III: 40.1–60%. Repellency class IV: 60.1– 80%; Repellency class V: 80.1–100%. a For RD50, Arabic numerals in parentheses are lower and upper fiducial limits, respectively.
et al. 1997; Stamopolous et al. 2007; Mossi et al. 2011) have reported the insecticidal properties of botanicals rich in 1,8-cineole against insects causing damage to stored products. 3. Experimental 3.1. Experimental site The experiment was carried out at the Storage Entomology Unit of Crop and Environmental Protection Laboratory, Ladoke Akintola University of Technology (LAUTECH), Ogbomoso, Nigeria. 3.2. Collection and handling of millet seeds Pearl millet seeds harvested in January 2011 from Bida, Niger State, Nigeria, were obtained from Wazobia Market, Ogbomoso, Nigeria, in April 2011 and were disinfested by storage at 48C for 2 weeks before use. Thereafter, the millet seeds were placed on laboratory bench for 2 days to equilibrate with the ambient relative humidity before use for the experimental purpose. 3.3. Culturing of T. castaneum T. castaneum was obtained from a culture maintained in Storage Entomology Unit of Crop and Environmental Protection Laboratory, LAUTECH, Ogbomoso, Nigeria. Insect rearing was done in the laboratory under ambient temperature of 25– 358C and relative humidity of 68 –80%, according to the method of Babarinde and Ogunkeyede (2008), using a portion of the millet seeds obtained from Wazobia Market, Ogbomoso. 3.4. Collection and extraction of EO from H. opposita leaves The leaves of H. opposita used for this study were obtained from Akoda Village, via Oyo, Nigeria, in September 2011. Authentication of the plant was done in the Department of Pure and Applied Biology, LAUTECH, Ogbomoso, where voucher specimen was dropped and a voucher number, LH0 368, was allocated. The plant material was air dried under shade in the laboratory until crisp, and later milled into fine powder using a milling machine. Two hundred and fifty grams of the powder was pre-soaked in 1000 mL of distilled water for 24 h, after which EO was obtained via
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hydrodistillation for 3 h with the aid of Clevenger apparatus according to the method recommended by British Pharmacopoeia (1988). The oil yield was 0.54% v/w. The EO obtained was stored in an air-tight glass sample bottle and kept in refrigerator until use. 3.5. Contact toxicity test Ten T. castaneum adults (1- to 7-day old) were selected for this bioassay. The contact toxicity bioassay followed the earlier method described by Babarinde and Ewete (2008) with some modifications. H. opposita EO at the rate of 1.0 mL was applied on the tergum of each insect with the aid of Hamilton syringe (Model 705 N, Sigmaw, St. Louis, USA). After the application of EO, the 10 insects were placed in 9 cm-diameter Petri dish and covered. A control experiment which was a collection of 10 T. castaneum adults similar in all respects as those used for the EO treatment was included. They were placed in 9 cm-diameter Petri dish and covered, without any treatment. The experiment was replicated three times. Mortality data were taken at 1, 24, 48, 72, 96, 120, 148 and 168 HAT. Insects that did not respond to pin probe were adjudged dead. Where there was mortality in the control, mortality was corrected with Abbott’s formula (Abbott 1925). Percentage mortality (PM) was calculated using the formula: PM ¼
Number of dead insects £ 100: Total number of experimented insects
3.6. Repellency test The method used for testing the repellency of H. opposita EO extract against T. castaneum adults was based on the area-preferred test described by Babarinde and Adeyemo (2010) with the following modifications. EO was applied at 10, 15, 20, 25 and 30 mL per half disc of (30 cm2) Whatman No. 1 filter paper in 9 cm-diameter Petri dishes, which corresponded to 0.33, 0.50, 0.66, 0.83 and 1.0 mL/cm2. The other half was left untreated. Control with 0.2 mL ethanol was also set up, with the other half of the filter paper left untreated. After 15 min of treatment of filter papers, 10 T. castaneum adults were introduced at the centre of the repellency bioassay chamber. There were four replicates for each treatment. Data on the repellence of EO against T. castaneum were taken at 1, 3 and 24 HAT. PR was calculated using the formula: PR ¼
Nu 2 Nt £ 100; Nu þ Nt
where Nu is the number of insects on untreated filter paper disc and Nt is the number of insects on treated filter paper disc. 3.7. Experimental design and data analysis The experiment was set up in completely randomised design. Data were transformed using angular transformation and subsequently subjected to analysis of variance (ANOVA) using SAS Package (SAS 2000). Where there was significant treatment effect, Tukey’s HSD test at 5% probability level was used to separate the means. Probit analyses were done to determine LT50 and RD50 for contact toxicity and repellency bioassays, respectively. 4. Conclusion The bioactivity of H. opposita EO against T. castaneum reveals its potentials to control the insect. This discovery can provide leads for active biopesticides for the protection of pearl millets against T. castaneum.
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Acknowledgement The authors acknowledge Dr A. T. J. Ogunkunle of Department of Pure and Applied Biology, LAUTECH, Ogbomoso, Nigeria, for authentication of H. opposita.
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References Abbott WS. 1925. A method for computing the effectiveness of an insecticide. J Econ Entomol. 18:265–267. Abdel-Sattar E, Zaitoun AA, Farag MA, El Gayed SH, Harraz FMH. 2010. Chemical composition, insecticidal and insect repellent activity of Schinus molle L. leaf and fruit essential oils against Trogoderma granarium and Tribolium castaneum. Nat Prod Res. 24(3):226–235. Adedire CO. 2003. Use of nutmeg, Myristica fragrans (Houtt) powder and oil for the control of cowpea storage bruchid, Callosobruchus maculatus (Fabricius). J Plant Dis Prot. 109:193–199. Athanassiou CG, Kavallieratos NG, Evergetis E, Katsoula AM, Haroutounian SA. 2012. Insecticidal efficacy of silica gel with Juniperus oxycedrus ssp. oxycedrus (Pinales: Cupressaceae) essential oil against Sitophilus oryzae (Coleoptera: Curculionidae) and Tribolium confusum (Coleoptera: Tenebrionidae). J Econ Entomol. 106 (4):1902–1910. Ayensu ES. 1978. Medicinal plants of West Africa. Algonac, MI: Reference Publications, 282 p. Babarinde SA, Adeyemo YA. 2010. Toxic and repellent properties of Xylopia aethiopica (Dunal) A. Richard on Tribolium castaneum Herbst infesting stored millets, Pennisetum glaucum (L.) R. Br. Arch Phytopathol Plant Prot. 43(8):810–816. Babarinde SA, Ewete FK. 2008. Comparative bioactivity of three Khaya species (Meliaceae) on Callosobruchus maculatus Fabricus (Coleoptera: Bruchidae). J Entomol Res Soc. 10(1):27–35. Babarinde SA, Ogunkeyede AF. 2008. Fecundity suppression of two botanicals on rust red flour beetle, Tribolium castaneum infesting sorghum. J Ultra Scientist Phys Sci. 20(2):411–413. Babarinde SA, Olabode OS, Akanbi MO, Adeniran OA. 2008. Potential of Tithonia diversifolia with Pirimiphos-methyl in control of Sitophilus zeamais (Coleoptera: Curculionidae). Afr J Plant Sci Biotechnol. 2(2):77–80. Babarinde SA, Oyegoke OO, Adekunle AE. 2011. Larvicidal and insecticidal properties of Ricinus communis seed extracts obtained by different methods against Tribolium castaneum Herbst (Coleoptera: Tenebrionidae). Arch Phytopathol Plant Prot. 44(5):451–459. British Pharmacopoeia. 1988. British Pharmacopoeia. Vol. 2. London: Her Majesty’s Stationery Office (HMSO); p. 137– 138. Gelfand M, Mavis D, Drummond RB, Ndemera N. 1985. The traditional medical practictioners in Zaimbawe, the principles, practice and pharmacopoeia. Gweru: Mambo Press. Haines CP. 1991. Insects and arachnids of tropical stored product their biology and identification. 2nd ed. Chatham, Kent: Natural Resources Institute. Hill DS. 1990. Pest of products and their control. London: Belhaven Press. Jibilou R, Amri H, Bouayad N, Ghailani N, Ennabili A, Sayah F. 2008. Insecticidal effects of extracts of seven plant species on larval development, a-amylase activity and offspring production of Tribolium castanuem (Herbst) (Insecta: Coleoptera: Tenebrionidae). Bioresour Technol. 99:959– 964. Mondal M, Khalequzzaman M. 2009. Ovicidal activity of essential oils against red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae). J Biosci. 17:57–62. Morton JK. 1963. In: Hutchinson J, Dalziel JM, editors. Flora of West Tropical Africa. 2nd ed. London, 450 p. Crown Agents. Mossi AJ, Astolfi V, Kubiak G, Lerin L, Zanella C, Toniazzo G, de Oliveira D, Treichel H, Devilla IA, Cansian R, Restello R. 2011. Insecticidal and repellency activity of essential oil of Eucalyptus species against Sitophilus zeamais Motschulsky (Coleoptera, Curculionidae). J Sci Food Agric. 91:273–277. Mujovo SF, Husseina AA, Marion Meyer JJ, Fourie B, Muthivh T, Lall N. 2008. Bioactive compounds from Lippia javanica and Hoslundia opposita. Nat Prod Res. 22(12):1047– 1054. Ngadjui BT, Tsopmo A, Ayafor JF, Connolly JD, Tamboue HH. 1995. New Pyrone substituted flavonoid from Hoslundia opposita. J Nat Prod. 58:109–111. Obeng-Ofori D, Reichmuth CH, Bekele J, Hassanali A. 1997. Biological activity of 1,8-cineole, a major component of essential oil of Ocimum kenyense (Ayobangira) against stored product beetles. J Appl Entomol. 121:237– 243. Obeng-Ofori D, Reichmuth CH, Bekele AJ, Hassanali A. 1998. Toxicity and protectant potential of camphor, a major component of essential oil of Ocimum kilmanscharicum, against four-stored product beetles. Int J Pest Manage. 44:203–209. Rai KN, Alur AS, Reddy CR, Kalapande HV, Birajdar SN, Reddy KMV, Reddy AR. 2007. Pearl millet production technologies for Andhra Pradesh and Maharastra. Global theme on crop improvement. Andhra Pradesh: International Crops Research Institute for the Semi-Arid Tropics, 16 p. SAS Institute. 2000. Statistical analytical systems SAS/STAT user’s guide version 8(2). Cary, NC: SAS Institute.
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Downloaded by [University of Southern Queensland] at 21:08 09 October 2014
Stamopolous DC, Damos D, Karagianidou G. 2007. Bioactivity of five monoterpenoid vapours to Triboluim confusum (du Val) (Coleoptera: Tenebrionidae). J Stored Prod Res. 43:571–577. Turaki JM, Sastawa BM, Kabir BGJ, Lale NES. 2007. Susceptibility of flours derived from various cereal grains to infestation by the rust-red flour beetle (Tribolium castaneum Herbst) (Coleoptera: Tenebrionidae) in different seasons. J Plant Prot Res. 47(3):279–288. Usman LA, Zubair MF, Adebayo SA, Oladosu IA, Muhammad NO, Akolade JO. 2010. Chemical composition of leaf and fruit essential oils of Hoslundia opposita Vahl grown in Nigeria. Am Eurasian J Agric Environ Sci. 8(1):40–43.
