Insecticidal Efficacy of Wild Medicinal Plants, Dhatura alba and Calotropis procera, against Trogoderma granarium (Everts) in Wheat Store Grains.
Trogoderma granarium (Everts) is one of the world's most destructive pests of cereals, oilseed, malting, and other foodstuffs. Current studies were aimed to investigate the effect of essential oils of two different medicinal plants (Dhatura alba and Calotropis procera) against two different strains of T. granarium. Plant extracts were obtained by Rotary Shaker apparatus by using acetone as solvent. Four different concentrations 5%, 10%, 15% and 20% were prepared by diluting in acetone. The data regarding mortality, growth regulation and repellency of T. granarium was observed.
Repellency of plant extracts was tested using area preference method while for the growth regulatory potential of plant extracts, the adults insects were released in wheat grains treated with different concentrations of plant extracts. All the experiments were carried out in an incubator at 30+-2AdegC and 65+-5% R.H using Completely Randomized Design (CRD). The results for repellency remained more significant for C. procera extract as compared to D. alba. While for growth the C. procera showed more significant differences. In case of mortality both D. alba and C. procera were found equally effective against T. granarium.
Medicinal plants, Biopesticides, Khapra beetle, Store grains.
About 90% grains formed for regular use of human and animals come from cereals like rice, wheat and maize (McFarlane, 1989), whereas insect pests are of an excessive worth for highest causes of loss of grains throughout the storage (Scotti, 1978). Khapra beetle (Trogoderma granarium) is one of the most destructive insect pests of stored grains. It is economically important pest causing both qualitative and quantitative losses due to its huge population, in addition to changing the grain into frass and damages may be too much (up to 70%). Thus it is necessary to control this pest by using suitable methods like suing biotic and a-biotic factors, but the most usable and famous/common method for controlling is use of insecticides (Haq et al., 2014). But the problem comes up in way that this pest creates resistance against insecticides and damage remains same instead of control.
The notable reasons of resistance are survival of this pest long time without food in minimal moisture contents and its cryptic nature of the larvae that live in cervices and cracks (Eliopoulos, 2013). Larvae of T. granarium cause severe damage after come out from diapause when it gets suitable conditions. The damaged seeds become unhealthy and found unfit for human use (Arain et al., 2006). Trogoderma granarium infested grains lost their market value, because larvae during feeding consume quite a lot of nutrients (Jood and Kapoor, 1993). The series of losses take place due to infestation of T. granarium over a period of 1 to 10 months in the range from 0.2 to 2.9% (Irshad et al., 1988).
Many practices have been evaluated to manage this pest including chemicals as most common and cheap method but the chemicals are not environment friendly and have bad impacts on grain quality and leave many poisonous ingredients in grains (Champ and Dyte, 1977; Georghiou and Tejeda, 1991). Beside their harmful effects toward human health, these chemicals have been noticed to be resistance against stored grain insect pests so, required to be replaced with some alternate sources of control. The numerous plants are neem, eucalyptus, dhatura, chrysanthemum, moringa, ginger, capsicum and many other weeds like Aak weed and many other naturally occurring plants are included. It is need of the day that, the reduced use of hard chemistry insecticides and their integration with botanicals and other softer chemistries may be included in IPM modules (Khan et al., 2014; Khaliq et al., 2014).
The representative samples of T. granarium from different geographical region are found to be sensitive to esfenvelarate (synthetic pyrethroid). However it is dependent on enzymatic activities and doses of applied chemicals (Shakoori et al., 2018). Currently a wide range of botanicals have been discovered which are eco-friendly and have no side effects on grain's life and human health (Regnault-Roger et al., 1993; Park et al., 2003). Plant source are getting better attention as prophylactic measures for the control of pests in storage, due to safety to other than target pests (Jood et al., 1996). Insecticidal effects of many plants beside pests of stored grains have been established. Many plants show their physiological and behavioral activities like; contact, repellent and antifeedant, against pests (Grainge and Ahmed, 1998).
Botanical insecticides are one of the most imperative groups of harmless and effective insecticides which are minus toxic to mammalian and are eco-friendly. Naturally occurring botanicals and medicinal plants are found to be cheaper, easily available and quite incorporable in the integrated-pest management packages. Moreover, the industrialized nations of the world are also stressing upon the adoption of organic farming for the preservation of the world health as well as for the development of sustainable agriculture. Therefore, it was important to exploit some botanicals for the organic control of stored-food insect pests. The current study was aimed to investigate two strains (Bhakkar and Faisalabad) of Trogoderma granarium by using Dhatura alba and Calotropis procera to observe their repellency, growth regulation and mortality of T. granarium.
MATERIALS AND METHODS
The present study was carried out in the Grain Research, Training and Storage Management Cell (GRTSMC) of the Department of Entomology, University of Agriculture Faisalabad during 2015-2016.
Collection of the insects
The adults and larvae of T. granarium were collected from grain market of Faisalabad district and were reared in laboratory to get homogenous population for further use in experiments.
Rearing and colony maintenance of insects
The insect culture was maintained in earthen pots (2.5 kg capacity) and plastic containers (1.5 kg). The pots were covered with organdy cloth, tightened with rubber bands to avoid the escape of insects. The insects were allowed to multiply in natural environment as in traditional structure and observed at regular intervals. The food medium was sterilized in an oven for 28 h at 60AdegC and thoroughly washed thereafter with water to remove any contamination or other insects present in the materials and carefully dried under sunlight, ensuring 13-14 % moisture content. The sterilized food was then preserved in air tight glass jars (1000 ml) in order to impede further infestation. Newly emerged 3rd instar larvae of T. granarium were used in the experiments.
Collection and preparation of botanical extract
The leaves of Dhatora alba, and Calotropis procera were collected from range land areas of district Bhakkar (Punjab). The leaves of tested plants were washed out using tape water. After shade drying the leaves were grinded in electrical grinder to bring these in the form of powder. The leaf extraction was made using Soxhlet's Extraction Apparatus by dipping 50 g of powder in 250 ml acetone, following procedure as described by Valladares et al. (1997) and Ahmad et al. (2006). Chemical extracts obtained were saved in clean and airtight lid glass flasks. The samples were stored in the refrigerator at 4AdegC for further use.
From the stock solution, various concentrations (5, 10, 15 and 20%) of each extract were prepared by using acetone as a diluting solvent. The prepared concentrations of these extracts were applied on sterilized wheat. After the evaporation of the solvent the air dried wheat (200g) was transferred into the treatment jars. The bioassays for each treatment were replicated thrice.
For repellency bioassay, same concentrations were applied on filter paper and twenty larvae of T. granarium were used for each treatment. The data was observed with the interval of 24 h and percent repellency was calculated. For insecticidal bioactivity and growth regulatory studies 30 grubs were released in sterilized glass jars containing wheat grains. Data for percent mortality was recorded with the interval of 24, 48 and 72 h and data regarding larval duration, percent pupation and percent adult emergence were also recorded with same pattern.
Before the analysis of data, normality was assessed using Kolmogrorov-Smirnov normality tests. Parametric tests were performed on normal data. The effect of different concentrations and plant extracts were analyzed with ANOVA using GLM and means were compared with Tuckey's HSD test at 5%. All statistical analyses were performed using computer software Stastix 8.1A(r).
The mean mortality for D. alba in Bhakkar strains was maximum (56.96) at 20% concentration and the minimum mortality (14.71) at 5%. Which were statistically different from other two concentrations 10% and 15% having mortalities (24.27) and (38.83), respectively. While C. procera showed maximum mortality (55.96) at 20% concentration and the minimum mean mortality (12.71) at 5%. Which were statistically at par with concentrations 10% having mean mortality (24.27) and 10% concentration was statistically at par 15% concentration having mortality (42.38). The mean mortality of Faisalabad strains according to different concentrations of D. alba the maximum mean mortality (57.44) was found at 20% concentration and the minimum mean mortality (15.44).
Which were statistically different from other two concentrations 10% and 15% having mortalities (33.16) and (42.87) respectively but these two concentrations was not significantly different. While the data regarding mean mortality according to different concentrations of C. procera in Faisalabad strains the maximum mean mortality (57.44) was found at 20% concentration and the minimum mean mortality (15.84). Which were statistically at par with concentrations 10% having mean mortality (33.16) and 10% concentration was statistically at par 15% concentration having mortality (42.87) (Fig. 1).
The mean repellency of different concentrations of D. alba in Faisalabad strains was significantly differed from each other. The highest repellency showed by the 5% concentration which's mean was 41.49 followed by 15% concentration with mean value of 25.76 and at 10% concentration mean repellency was 40.97and at 20% concentration the mean repellency was 16.02. While the mean repellency of different concentrations of C. procera was significantly differed from each other. The highest repellency showed by the 5% concentration with mean of 51.49 followed by 15% concentration with mean value of 35.76 and at 10% concentration mean repellency was 50.97 and at 20% concentration the mean repellency was 26.02. The mean repellency of different concentrations of D. alba in Bhakkar strains was significantly differed from each other.
The highest repellency showed at 20% concentration (mean 51) followed by 15% concentration (mean 50), at 10%, 5% concentration the mean repellency was 35 and 26 respectively. However, the mean repellency of different concentrations of C. procera was significantly differed from each other. The highest repellency showed at 20% concentration (mean 50) followed by 15% concentration (mean 34), at 10%, 5% concentration the mean repellency was 48.33 and 25, respectively (Fig. 1).
The mean mortality by D. alba in Bhakkar strains was observed for different time intervals. Maximum mean mortality (36.93) was found in 72 h while exposure time 24 h gave minimum mean mortality (25.2). Which have no significantly differed to other time interval that was 48 hours having mean mortality (32.5); while for C. procera the maximum mean mortality (32.53) was found at 72 h of observation, while exposure time 24 h gave minimum mean mortality (25.28). Which have significant difference with other time interval that was 48 h having mean mortality (36.93). Data regarding the mean mortality by D. alba in Faisalabad strains observed for different time intervals. Maximum mean mortality (44.48) was found in 72 h while exposure time 24 h showed minimum mean mortality (37.33). Which have significantly no difference with other time interval i.e. 48 h having mean mortality (37.72).
For C. procera the mean mortality was maximum (48.28) was found at 72 h while exposure time 24 h provided minimum mean mortality (25.54). Which have significant difference with 48 h observations having mean mortality (38.18) (Fig. 2).
Table I.- Percentage of pupation and adult eclosion of T. granarium by different concentrations of D. alba and C. procera extracts.
Treatments/###Bhakkar strain (grains)###Faisalabad strain (grains)
Stage/###D. alba###C. procera###D. alba###C. procera
###(%)###eclosion (%)###(%)###eclosion (%)###(%)###eclosion (%)###(%)###eclosion (%)
The mean percent pupation by D. alba in Bhakkar strains of different concentrations was significantly differed from each other. The highest percent pupation showed @ 5% concentration (mean 77.78%) followed by 10% concentration (mean 65.56%) and @ 15% , 20% concentration the percent pupation was 57.78% and 51.11%, respectively; while the mean percent pupation by C. procera of different concentrations was significantly differed from each other. The highest percent pupation showed @ 5% concentration (mean 83.33%) followed by 10% concentration (mean 75.56%) and @ 15%, 10 % concentration the percent pupation was 66.67% and 56.67%, respectively. Same trend was also found for D. alba and C. procera in Faisalabad strains. For both botanicals i.e. D. alba and C. procera and in two different strains of T. granarium at different concentrations were significantly differed from each other as shown in Table I.
The maximum repellency showed in Bhakkar strains for extracts comparisons of D. alba and lesser was observed when treated with different concentrations of C. procera. For time intervals check mortality the maximum repellency showed by D. alba. The current observations are showing that with increased concentrations, the repellency also increased and vice versa. The assorted behavioral and physiological effects such as repellency shown in this study, and toxicity, oviposition deterrence and reproduction inhibition are in agreement with Bodnaryk et al. (1999) who suggested the active principles may constitute a complex of chemicals while applications are done with different concentrations and observed at different time intervals. Delobel et al. (1999) have isolated a 37 amino acid protein from pea seed that are toxic to Sitophilus spp.
However, they have not determined if it is also repellent to stored-product insects. For growth regulation of T. granarium a 70 days process was done for both strains. The results for Bhakkar strain were maximum for C. procera and also in Faisalabad strain but different in counts, which are concede with those of El-Nadi et al. (2001) and Al-Moajel (2004).
It is concluded that the tested biochemicals may be utilized as alternative to hard chemistries in insect control modules of stored grains. In addition to that, their less persistency may lead to safe protection of stored products.
Statement of conflict of interest
Authors have declared no conflict of interest.
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|Publication:||Pakistan Journal of Zoology|
|Date:||Feb 28, 2019|
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