Brazilian Journal of Biological Sciences, 2016, v. 3, no. 6, p. 367-374. ISSN 2358-2731 http://dx.doi.org/10.21472/bjbs.030612

Mothcidal activity of three botanical extracts against Sitotroga cerealella (Olivier, 1789) (Lepidoptera: Gelechiidae) infesting rice grains Olajire Ayodele Gbaye, Emmanuel Ayobami Oyeniyi* and Janet Titilayo Eweniyi

Department of Biology. Federal University of Technology. P. M. B. 704. Akure. Nigeria. Email: [email protected]; [email protected].

Abstract. The renewed awareness on the potential adverse effects associated with several synthetic insecticides has continually served Received as impetus to search for safer and more sustainable pesticides August 26, 2016 derived from plant origin. In this study, the mothcidal activity of Alstonia boonei (de wild), Garcinia kola (Heckel) and Accepted December 22, 2016 Crassocephalum crepidoides (Benth) against Sitotroga cerealella (Olivier, 1789) was investigated at ambient temperature (28 ± 2 °C) Released and relative humidity (75% ± 5%). Adult moths were exposed to December 31, 2016 1%, 2% and 3% concentration of each oil extract and mortality was recorded at day 1, 2, 3, 4 and 5 post-treatment, respectively. The Open Acess Full Text Article control experiments were also set-up. All the three botanicals exhibit moth mortality irrespective of the concentration and exposure time. However, only C. crepidoides elicited complete moth mortality (100%) irrespective of the concentration and exposure time while A. boonei oil extract applied at 1%, 2% and 3% showed the least moth mortality regardless of the exposure time. Similarly, 2% and 3% concentration of each botanical showed complete moth mortality at 4 and 5 days post-treatment, respectively. Least (6.60%) and highest (23.30%) adult emergence was also observed in grain exposed to 1% of C. crepidoides and A. boonei, respectively, while G. kola and C. crepidoides applied at 3% completely prevented ORCID moth emergence and loss in weight in rice grains. This study showed 0000-0002-2988-5223 that oil extract of G. kola and C. crepidoides would provide a much Olajire Ayodele Gbaye desired wherewithal to synthetic insecticides in the management of 0000-0003-2319-8091 S. cerealella infesting rice grains in Nigeria. Emmanuel Ayobami Oyeniyi 0000-0001-8623-4438 Keywords: Alstonia boonei; Garcinia kola; Crassocephalum Janet Titilayo Eweniyi crepidoides; Sitotroga cerealella; mothcidal.

Introduction infestation by insect pests such as weevils, beetles and moths. The inability of the In Nigeria, one of the most critical country to achieve self-sufficiency in food expectations of the populace from the production is further aggravated by her government is the need to increase the over-reliance on oil proceeds which has led nation’s domestic food production; as the to the negligence of other sectors of the little food being produced are not enough, nation’s economy for several years. The and still suffers from poor processing recent dwindling in the nation’s income due techniques which pre-dispose them to to incessant drop in global oil price has

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further necessitated the need to diversify the as a substitute to most synthetic chemicals. nation’s economy away from oil This has led to the usage of several plant dependency. Agriculture has been materials in the control of S. cereallela advocated as one of the alternative (Zaidi et al., 2004; Iqbal, 2010; Ashamo wherewithal that could serve as the and Akinnawonu, 2012). However, high mainstay of the nation’s economy (Adubi, abundance of prospective insecticidal floras 2004). This has led to the establishment of in Nigeria has given several researchers the various Agricultural Research Institute all impetus to constantly investigate more over the country saddled with the floras for their efficacy against this responsibility of developing production and infamous pest of rice. This is needed for the utilization technologies for various crops in actualisation of the nation’s much desired Nigeria (Olatunde, 2014). Notwithstanding, self-sufficiency in rice production. Thus, most Nigeria farmers and crop merchants the mothcidal efficacy of Alstonia boonei still record huge post-harvest losses due to (de wild), Garcinia kola (Heckel) and low level of literacy on modern storage Crassocephalum crepidoides (Benth) techniques that could guarantee food against S. cerealella infesting rice grains security in Nigeria. was investigated in this study. Rice is one of the cereal crops that could contribute to the realisation of food Materials and methods security in sub-Sahara Africa. Nigeria remained the largest producer, consumer Insect culture and importer of rice in West-Africa with a The infested paddy stock culture of local production of about 4.2 million tonnes S. cerealella and clean paddy rice variety (FAO, 2013). The crop plays a major (FARO-52) used in this study were nutritional role in the diet of most Nigerians obtained from Agricultural Development and also forms an important part of their Project (ADP), Akure, Ondo State, Nigeria. daily nourishment. Nonetheless, the country Clean paddy rice variety (FARO-52) was still struggles to achieve self-sufficiency in disinfested in the freezer at -18 oC for two rice production due to various limitations of weeks and thereafter allowed to equilibrate which insect pest infestation during storage in the laboratory for three days. 200 g of served as one of the most important disinfested clean paddy rice variety were constraints. Sitotroga cerealella (Olivier) introduced into 1.5 L plastic containers and has been implicated as one of the moths from starter culture were later reared destructive storage pest militating against on the paddy rice variety. The containers rice production in Nigeria (Ashamo and were covered with perforated lid and muslin Akinnawonu, 2012). Synthetic chemicals cloth to prevent the escape of insects and however remained the most effective allow air into them. The moths were reared method of managing stored product pest in for two generations on the paddy rice the country (Chedi and Aliyu, 2010). variety to eliminate the effect of parent food However, recent ban by the European before being used for bioassay test. The Union on some agricultural food export plastic containers used for insect rearing from Nigeria due to high level of pesticides were kept inside cages at ambient has necessitated the need for a better and temperature of 28 ± 2 ºC and 75% ± 5% sustainable means of protecting farm relative humidity. produce during storage. Likewise, the various adverse environmental, biological Procurement and processing of and health consequences associated with plant materials most synthetic chemicals have further The three plant materials used in prompted the urgent need for better control this study were obtained from three measures. different sources. Leaves of G. kola and The search for effective and eco- C. crepidoides were sourced from Aba Oyo friendly means of pest control on rice grains Village and Odopetu Market, Akure, Ondo has led to the touting of several botanicals State, respectively. Stem bark of A. boonei

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Mothcidal activity of three botanical extracts against Sitotroga cerealella 369

was however obtained from Fayemi Twenty grams of paddy rice Market, Ado-Ekiti, Ekiti State. Fresh leaves (FARO-52) seeds were weighed into and stem bark of various plants were dried 170 mL plastic containers using Metler in the laboratory using a dry cabinet at a beam PB 3002 weighing balance and 1 mL temperature of 40 °C and later pulverized of each plant extract concentrations of 1%, separately using NAKAI NJ-1731 electric 2% and 3% was separately mixed with the blender. Powders of each plant material paddy rice. Each container was left for 1hr were later sieved with a mesh size of 1 mm² to allow the methanol solvent to vapourise before being separately stored in plastic leaving only the oil extracts of each containers with airtight lids for subsequent botanical on the seeds. Two controls were use. set up for each plant material, one with To prepare the extracts, Metler solvent alone (Control A) and the other beam weighing balance was used to weigh with neither solvent nor extract (Control B). twenty grams of each pulverized plant Air space was created by cutting the centre material into a muslin cloth. The muslin portion of the lid of each container and cloth containing each botanical was later replacing with muslin cloth. Five pairs of transferred into the thimble and extracted adult moths (0-24 h old) were introduced with methanol in a soxhlet apparatus. The into each plastic container using aspirator. extraction was carried out for 3-4 h Each treatment was replicated three (3) depending on the plant material. The times and adult mortality was recorded extraction was terminated when the solvent daily for 5 days. Both dead and live insects in the thimble became clear. The thimble were removed on the sixth day and the was removed from the unit and the solvent experiments were left for 30 days to allow recovered by distilling in the soxhlet for adult emergence and the number of extractor. The resulting extracts contained emerged adults was counted. Inhibition rate both the solvent and the oil. The solvent (%IR) in adult emergence was calculated was separated from the oil using rotary using the method described by Tapondju et evaporator, after which the oil was exposed al. (2002). to air so that traces of the volatile solvents evaporate, leaving the oil extract. This is important for the avoidance of false concentrations. From this main stock solution, Where Cn is the number of insects different concentrations of 1%, 2% and 3% that emerged in the control treatment and Tn oil concentrations were made. A is the number of adult insects that emerged concentration of 1% was made by diluting in the treated grains. The weight loss of the 0.1 mL of plant extract in 9.9 mL of stored grains after 30 days was calculated methanol (solvent). Two percent (2%) using the formula below: concentration was made by diluting 0.2 mL of plant extract in 9.8 mL of methanol while 3% concentration was made by diluting 0.3 mL of each plant extract with 9.7 mL of methanol. The various Statistical analysis concentrations were made using small glass All data on adult mortality were bottles and graduated syringes. After each checked for normality based on Shapiro- dilution, the syringe was rinsed with the Wilk Test before being subjected to one- solvent while different syringes were used way analysis of variance (ANOVA) for different plant material extracts. (p < 0.05) and treatment means were separated using Tukey’s Test. Analyses Effect of oil extracts on mortality, were carried out using SPSS 17.0 software adult emergence, inhibition rate (%IR) package. and grain weight loss

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370 Gbaye et al.

Results Significantly higher (P > 0.05) moth mortality was also observed in moth treated Effect of oil extract on the adult with botanicals when compared to their mortality of S. cereallella counterpart in control. Solvent treated Table 1 to 3 show the effect of 1%, control also showed 10% moth mortality 2% and 3% oil extract of A. boonei, G. kola after 5 days post-treatment (Tables 1 to 3). and C. crepidoides on adult mortality of Least moth mortality was also observed in S. cereallella exposed for 5 days. Of all the A. boonei oil extract applied at 1%, 2% and botanicals, C. crepidoides elicited complete 3%, respectively, regardless of the exposure moth mortality (100%) irrespective of the time (Tables 1 to 3). Significant differences concentration and exposure time. Similarly, however existed in the mortality of moths 2% and 3% concentration of each botanical exposed to 1% and 2% concentration of showed complete moth mortality at 4 and 5 each botanical after day 1 (24 h post- days post-treatment, respectively. treatment) (Table 1 and 2).

Table 1. Effect of 1% oil extracts of the three botanicals on the percentage mortality of adult S. cerealella.

Duration Botanical 1 2 3 4 5 A. boonei 36.70 ± 0.33b 46.70 ± 0.88b 60.00 ± 0.58b 83.30 ± 0.88b 96.70 ± 0.33c G. kola 56.70 ± 0.96c 86.70 ± 0.67c 100 ± 0.00c 100 ± 0.00c 100 ± 0.00c C. crepidoides 100 ± 0.00d 100 ± 0.00 100 ± 0.00c 100 ± 0.00c 100 ± 0.00c Control A 0.00 ± 0.00a 0.00 ± 0.00a 0.00 ± 0.00a 0.00 ± 0.00a 10.00 ± 0.58b Control B 0.00 ± 0.00a 0.00 ± 0.00a 0.00 ± 0.00a 0.00 ± 0.00a 0.00 ± 0.00a Means followed by the same letter along the column are not significantly different (P > 0.05) using Tukey Test.

Table 2. Effect of 2% oil extracts of the three botanicals on the percentage mortality of adult S. cerealella.

Duration Botanical 1 2 3 4 5 A. boonei 46.70 ± 0.33b 66.70 ± 0.88b 83.30 ± 0.67b 100 ± 0.00b 100 ± 0.00c G. kola 83.30 ± 0.88c 96.70 ± 0.33c 100 ± 0.00c 100 ± 0.00b 100 ± 0.00c C. crepidoides 100 ± 0.00d 100 ± 0.00c 100 ± 0.00c 100 ± 0.00b 100 ± 0.00c Control A 0.00 ± 0.00a 0.00 ± 0.00a 0.00 ± 0.00a 0.00 ± 0.00a 10.00 ± 0.58b Control B 0.00 ± 0.00a 0.00 ± 0.00a 0.00 ± 0.00a 0.00 ± 0.00a 0.00 ± 0.00a Means followed by the same letter along the column are not significantly different (P > 0.05) using Tukey Test.

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Mothcidal activity of three botanical extracts against Sitotroga cerealella 371

Table 3. Effect of 3% oil extracts of the three botanicals on the percentage mortality of adult S. cerealella. Duration Botanical 1 2 3 4 5

A. boonei 60.00 ± 0.00b 76.70 ± 0.33b 100 ± 0.00b 100 ± 0.00b 100 ± 0.00c G. Kola 96.70 ± 0.33c 100 ± 0.00c 100 ± 0.00b 100 ± 0.00b 100 ± 0.00c C. crepidoides 100 ± 0.00c 100 ± 0.00c 100 ± 0.00b 100 ± 0.00b 100 ± 0.00c Control A 0.00 ± 0.00a 0.00 ± 0.00a 0.00 ± 0.00a 0.00 ± 0.00a 10.00 ± 0.58b Control B 0.00 ± 0.00a 0.00 ± 0.00a 0.00 ± 0.00a 0.00 ± 0.00a 0.00 ± 0.00a Means followed by the same letter along the column are not significantly different (P > 0.05) using Tukey Test.

Effect of botanical extracts on the rate (Table 5). Of all the three botanicals, emergence and inhibition in adult A. boonei showed the least emergence emergence of S. cerealella (59.29-95.46%) while the highest inhibition Table 4 and 5 shows the effect of rate was observed in grains treated with oil extract of the three botanicals on the C. crepidoides oil extracts (91.04%-100%) adult emergence and inhibition in adult (Table 5). Median inhibition in adult emergence of S. cerealella exposed to emergence (72.86%-100%) was however concentrations of 1%, 2% and 3%, observed in grains treated with G. kola oil respectively. Significantly higher (P < 0.05) extracts (Table 5). adult emergence was observed in grains exposed to solvent treated control (Control A) (70.00) and ordinary control (Control B) (73.30) when compared to their counterpart Effect of oil extracts on weight exposed to botanical oils (Table 4). Among loss in rice grains the treated grains, least (6.60%) and highest The weight loss in rice grains (23.30%) adult emergence was observed in treated with different concentrations of oil grain exposed to 1% of C. crepidoides and extracts from the three botanicals is A. Boonei, respectively (Table 4). Also, prevented in table 6. Significantly higher G. kola and C. crepidoides applied at 2% (P < 0.05) weight loss was observed in and 3% completely prevented moth grains exposed to both solvent control emergence with the exception of G. kola (15.78) and ordinary control (18.63) when applied at 2% where emergence of 3.33% compared to weight loss in grains treated was observed (Table 4). with botanical extracts. At the highest Significantly lower (p < 0.05) experimental concentration (3%), C. percentage inhibition rate was observed in crepidoides and G. kola however adult emergence of grains exposed to both completely (0.00) prevented loss in weight control when compared to their counterpart in rice grains and this was significantly treated with botanical oil extracts (Table 5). different (P<0.05) from weight loss However, complete inhibition (100%) was observed in both control. Of all the observed in adult emergence of grains botanicals, least reduction (0.61-8.70%) in exposed to 2% and 3% oil extracts of weight loss was observed in grains exposed C. crepidoides and G. kola, respectively to A. boonei while the highest reduction (Table 5). The only exception was observed (0.00-2.10%) in weight loss was observed in grains exposed to 2% oil extract of in grains treated with C. crepidoides. G. kola which showed 95.46% inhibition

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Table 4. Effect of oil extracts of three botanicals on the adult emergence of S. cerealella. Concentration (%) Botanical 1 2 3 A. boonei 30.00 ± 0.00c 23.30 ± 0.33b 3.33 ± 0.33a G. kola 20.00 ± 0.37b 3.33 ± 0.33a 0.00 ± 0.00a C. crepidoides 6.60 ± 0.33a 0.00 ± 0.00a 0.00 ± 0.00a Control A 70.00 ± 0.57d 70.00 ± 0.57c 70.00 ± 0.57b Control B 73.70 ± 0.88d 73.33 ± 0.88c 73.30 ± 0.88b Means followed by the same letter along the column are not significantly different (P > 0.05) using Tukey Test.

Table 5. Effect of oil extracts of three botanicals on inhibition rate (%IR) in emergence of S. cerealella.

Concentration (%) Botanical 1 2 3 A. boonei 59.29 ± 0.00b 68.25 ± 0.21b 95.46 ± 0.12b G. kola 72.86 ± 0.13c 95.46 ± 0.12c 100 ± 0.00b C. crepidoides 91.04 ± 0.21d 100 ± 0.00c 100 ± 0.00b Control A 5.02 ± 0.03a 4.54 ± 0.00a 4.54 ± 0.00a Control B 0.00 ± 0.00a 0.00 ± 0.00a 0.00 ± 0.00a Means followed by the same letter along the column are not significantly different (P > 0.05) using Tukey Test.

Table 6. Effect of oil extracts of three botanicals on weight loss in rice grains.

Concentration (%) Botanical 1 2 3 A. boonei 8.70 ± 0.12c 4.91 ± 0.79b 0.61 ± 0.01a G. Kola 5.86 ± 0.37b 0.71 ± 0.71a 0.00 ± 0.00a C. crepidoides 2.10 ± 0.18a 0.00 ± 0.00a 0.00 ± 0.00a Control A 15.78 ± 0.04d 15.78 ± 0.04c 15.78 ± 0.04b Control B 18.63 ± 0.02e 18.63 ± 0.02d 18.63 ± 0.02c Means followed by the same letter along the column are not significantly different (P > 0.05) using Tukey Test.

Discussion synthetic chemicals. Alstonia boonei, Garcinia kola and Crassocephalum The renewed awareness on the crepidoides are some of the botanicals in potential adverse effects associated with Nigeria that have been established to several synthetic insecticides coupled with possess medicinal properties (Burkil, 2004; high demand for safer and more sustainable Odugbemi, 2006). agricultural practices has continuously This present study therefore serves as impetus for the use of pesticides investigates the mothcidal activity of oil derived from plant origin. Consequently, extracts of A. boonei, G. kola and C. most farmers in developing countries have crepidoides against S. cerealella infesting resorted to the use of botanicals for pest rice grains in Nigeria. Various results management to circumvent the high cost obtained showed that all the botanical oil and potential risks associated with most extracts resulted in moth mortality, reduced adult emergence and weight loss in grains

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Mothcidal activity of three botanical extracts against Sitotroga cerealella 373

irrespective of the exposure days and and efficiency in the exposed insect concentration when compared to control. (Willmer et al., 2000; Lattanzio et al., This shows that they all possessed 2005). This might have resulted in mothcidal activity against S. cerealella, starvation of S. cerealella larvae leading to thus corroborating the findings of Iqbal death and subsequent reduction in adult (2010), Ashamo and Akinnawonu (2012) emergence in grains exposed to oil extract and Akinneye and Oyeniyi (2016) where of C. crepidoides. Yang et al. (2006) has the entomotoxic effect of several botanical also reported that alkaloids and flavonoids pesticides against S. cerealella have been disturb and reduced larval growth and reported. The high mortality associated with survival which ultimately affect the life the oils of these botanicals may be ascribed cycle of adult insects. to disruption in the normal respiratory Oil extracts of the three botanicals activities of adult moths leading to the also significantly reduced weight loss in asphyxiation and subsequent death treated grains when compared to control. (Ashamo et al., 2013; Ogungbite and This shows that all the three botanicals Oyeniyi, 2014). could help in preserving rice grains in Moth exposed to grains treated with Nigeria. However, grains exposed to C. oil extract of C. crepidoides however crepidoides oil extract show the least showed the highest mortality when weight loss while their counterpart exposed compared to other botanicals. The ability of to A. boonei showed the highest weight the oil extract from C. crepidoides to elicit loss. The ability of C. crepidoides oil highest mortality may be accredited to the extract to prevent weight loss especially at pungent smell of this plant which may have the highest experimental concentration may increased its toxicity against adult moths. be attributed to the inability of the larvae of Of all the three botanicals, C. crepidoides S. cerealella to feed on the treated rice showed the highest pungent smell. Moths grains. Similar observation has been are known to be active fliers which may reported by Akinneye and Oyeniyi (2016) have increased the rate of diffusion of C. on rice grains treated with Cleistopholis crepidoides vapour through the insect patens (Benth). spiracles. This might have led to the blockage of moth spiracles, suffocation and Conclusion higher mortality observed in insects exposed to oil extract of C. crepidoides Various results obtained in this when compared to others. study showed that oil extract from C. The higher efficacy of the three crepidoides showed the highest mothcidal botanicals, particularly C. crepidoides oil activity of the three botanicals due to its extract in preventing adult emergence when ability to exhibit complete moth mortality, compared to control may also be ascribed to inhibit adult emergence and prevent weight some toxic phytochemical compounds that loss in treated grains at the highest have been reported in these plants. For experimental concentration while A. boonei instance, Oduse et al. (2012) reported the showed the least mothcidal activity. Due to presence of tannins, flavonoids and the non-toxic nature of C. crepidoides on alkaloids in C. crepidoides. Most of these humans as it is normally cooked and taken compounds had been reported for their as vegetable soup in Nigeria (Oduse, 2012); considerable toxicity and antifeedant effect further studies should be carried out on this towards insects (Yang et al., 2006). These botanical to establish the mothcidal active compounds might have dissolved in oils compounds. Crop merchants and peasant and form a protective coating on the paddy farmers should also be encourage to use oil rice. Tannins for example are known to extract from this plant in protecting their induce indigestion in insects by binding paddy rice against S. cerealella attacking with molecules such as protein and paddy rice in Nigeria. This could help in polysaccharides to form complexes which reducing high loss usually associated with could consistently reduce the assimilation

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