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Insecticidal Activity of Essential Oils of Four Medicinal Plants Against Different Stored Grain Insect Pests.

Byline: Shahzad Saleem Mansoor ul Hasan Muhammad Sagheer and Shahbaz Talib Sahi

Abstract

Studies were carried out to investigate the insecticidal activity of essential oils of four locally grown plants such as Datura stramonium Eucalyptus camaldulensis Moringa oleifera and Nigella sativa against three major insect pests viz. Tribolium castaneum Trogoderma granarium and Cryptolestes ferrugineus responsible for economic loss to stored commodities. Test insects were fumigated with concentrations of 5 10 15 and 20 l/L under laboratory conditions at 302C and 655% relative humidity. Essential oils fumigation significantly affected the mortality at all levels of concentration and exposure periods. Among essential oils D. stramonium was found to be the most toxic against T. granarium (14.46%) and C. ferrugineus (28.49%) while N. sativa showed the highest fumigant mortality (20.06%) against T. castaneum. Overall results show that C. ferrugineus was found to be the most susceptible test insect with 23.79 % mean mortality followed by T. castaneum (17.11%) and T. granarium (12.27%).

Higher mortality was found to be concentration and exposure time dependent. The results demonstrate that the essential oils of investigated plants can be used as fumigant to control insect pests of stored products.

Keywords: Plant essential oil fumigant toxicity Tribolium castaneum Trogoderma granarium Cryptolestes ferrugineus

INTRODUCTION

After harvesting approximately 1660 insect species attack the agricultural produce during different phases like transportation processing marketing and storage (Hagstrum and Subramanyam 2009). Losses caused by these insect pests may reach up to 30% during storage (Haubruge et al. 1997). Among these insect pests Tribolium castaneum (Mondal 1994; Danahaye et al. 2007) Trogoderma granarium (Burges 2008; Mark et al. 2010) and Cryptolestes ferrugineus (Suresh et al. 2001; Mason 2003) are documented to be the most damaging and destructive pests of stored products throughout the world. About 2-6% food grain production of Pakistan is lost every year during storage by stored grain insect pests (Avesi 1983)

Synthetic insecticides (pyrethroids and organophosphates) and fumigants (methyl bromide and phosphine) are commonly used to control these insect pests throughout the world. Out of these control strategies fumigants (because of their broad spectrum action and rapid penetration without residues) are known to be convenient and economical control measure (Mueller 1990; Varma and Dubey 2001; Ogendo et al. 2008). Methyl bromide is completely phased out as it was found one of main causes of ozone depletion (Butler and Rodriguez 1996; Shaaya and Kostyukovsky 2006; Tayoub et al. 2012). The control of stored product insect pests mainly depends on application of phosphine (Varma and Dubey 2001). There are also many problems associated with its application such as adverse effects on non target organisms and environment human health safety concerns and pest resistance and resurgence (Ogendo et al. 2008).

Almost all major pests of stored products have developed resistance against phosphine (Pimentel et al. 2007; Lorini et al. 2007; Nayak et al. 2012).

This situation demands a serious effort to find out some safe viable biodegradable environment friendly and effective substitute to these conventional fumigants. Botanicals extracted from higher plants have been found suitable after investigating their fumigant insecticidal properties by many scientists (Isman 2006 2008; Rajendran and Sriranjini 2008; Sagheer et al. 2013). A wide number of plant essential oils and their constituents have been proved for their fumigant insecticidal action against stored product pests (Kim and Ahn 2001 Singh et al. 2005; Opolot et al. 2006; Tripathi et al. 2009; Lopez and Pascual-Villalbos 2010).

In many studies Eucalyptus camaldulensis have shown its fumigant potential against T. castaneum (Tunc et al. 2000; Channoo et al. 2002; Negahban and Moharramipour 2007). Similarly Ogendo et al. (2008) performed space fumigation tests to evaluate bioactivities of O. gratissimum against five major pests of stored grain. T. castaneum and S. oryzae were found to be the most tolerant among the insects as they showed 23 and 94% mortality at 10 l/air for 168 hr of exposure respectively while R. dominica O. surinamensis and C. chinesis exhibited 98 99 and 100% mortality respectively even at 1l/air concentration with 24 h of exposure time.

Though effectiveness of volatile oils as fumigant has been well investigated but their activity at different concentration levels and exposure time against local strains of stored grain insect pests in Punjab Pakistan is yet to be evaluated.

MATERIALS AND METHODS

Plant materials

Leaves from the locally grown medicinal plants Datura stramonium (Dhatora) Eucalyptus camaldulensis (Sofaida) Moringa oleifera (Sohanjana) and seeds from Nigella sativa (Kalwanji) were collected from University of Agriculture Faisalabad Punjab Pakistan (Longitude 7374 East; Latitude 3031.5 North; Altitude: 184 m). These fresh plant parts were brought to laboratory (Grain Research Training and Storage Management Cell Department of Entomology University of Agriculture Faisalabad and dried in shade. Dried plant parts were grinded to powder using stone electric grinder (Machine No. 20069 Pascall Engineering Co. Ltd.). This grinded material was then sieved through 40 mesh sieve to obtain fine powder.

Extraction of essential oils

Soxhelt extraction apparatus (Model WHM12295 Daihan Scientific Co. Ltd.) was used to prepare essential oils. Soxhelt thimble was filled with 50 g of fine botanical powder and placed in flask. Acetone was used as solvent in bottom flask. This process of extraction oil from all plant powders was repeated many times to achieve enough quantity of essential oil. Extracted essential oil was then purified by evaporating solvent by using electric rotary evaporator. These pure extracted essential oils were preserved in glass vials at 4C and then used to prepare appropriate concentrations of 5 10 15 and 20 l/L by mixing acetone as solvent. These prepared concentrations were used for subsequent experiments.

Test insects

To collect three strains of Cryptolestes ferrugineus Tribolium castaneum and Trogoderma granarium Infested grain samples from three districts Faisalabad (FSD strain) (Longitude 7374 East; Latitude 3031.5 North; Altitude: 184 m) Sahiwal (SWL strain) (Longitude 73-06; Latitude 30-40 north; Altitude 150 m) and Toba Tek Singh (TTS strain) (Longitude 7208' to 7248; Latitude 3033' to 312 north; Altitude 162 m) of central Punjab Pakistan were collected and brought to laboratory. Populations of each test insect were sorted out as a strain from the collected samples of each district.

Collected insects were kept in the jars of 9.5cm diameter having commodity (wheat grains for T. granarium and wheat flour for C. ferrugineus and T. castaneum) sterilized for 30 min at 70C using oven (Lab Line Instruments Inc. Model no 3512-1) and covered with the muslin cloths. After three days adults were sieved out from the commodity. The rearing commodity along with eggs of the insects was put into jars which were kept at 302C and 655% relative humidity in incubator (MIR-254 Sanyo) to obtain F1 population which was considered as a homogenous culture (Mujeeb and Shakoori 2012). This homogenous culture was used for different bioassays using plant essential oils.

Bioassays

Space fumigation method (Shaaya et al. 1991) was used for the evaluation of fumigant toxicities of essential oils. 0.5-L flat-bottom air tight plastic space fumigation chambers with a small amount of food were used for experiment. Twenty adults of each test insect species were introduced in each test chamber. Essential oils were applied separately on small pieces of Whatman No. 1 filter paper to provide dosages of 5 10 15 and 20 l/L air. Control was treated with acetone alone. Treated filter papers were kept in air and allowed to evaporate the solvent (acetone) for 10 min. Fumigation chambers were sealed to make them air tight after suspending the treated filter papers in the middle of chamber below the lid to avoid physical contact and to assure uniform distribution of fumes. Data for adult mortality was recorded after 72 (3 days) 120 (5 days) and 168h (7 days) of treatment.

Statistical analysis

The data were arranged in tabulated format. The mortality (%) was corrected by Abbot's (1925) formula: Equation

The collected data were subjected to Analysis of Variance using Statistica software (8.0 Stat Soft Inc. 1984-2008). Means of significant treatments were separated using Tuckey-HSD test at a =5 %.

RESULTS

Three major insect pests of stored grains viz. T. granarium C. ferrugineus and T. castaneum were fumigated with essential oils of D. stramonium E. camaldulensis M. oleifera and N. sativa to evaluate fumigation potential of essential oils. Insecticidal efficacy of essential oils was found significant against all test insects (T. castaneum F(3436) = 96.761 p = 0.000; T. granarium F(3436) = 69.247 p = 0.000; C. ferrugineus F(3436) = 67.291 p = 0.000).Results revealed that maximum fumigation toxicity was depicted by D. stramonium against T. granarium (14.43 %) and C. ferrugineus (28.49 %) whereas against T. castaneum N. sativa was observed the most toxic (20.06 %) essential oil. There was not considerable difference found in fumigant toxicities of D. stramonium (14.34%) and E. camaldulensis (14.25%) against T. granarium.

While M. oleifera was found least toxic with mean mortality of 12.03 9.38 and 20.44% against T. castaneum T. granarium and C. ferrugineus respectively. According to susceptibility following order was observed in the test insects C. ferrugineus (23.79% mortality) greater than T. castaneum (17.11% mortality) greater than T. granarium (12.25% mortality).

Effect of concentrations of each essential oil on mortality of test insects was found significant (T. castaneum F(3436) = 346.40 p = 0.000; T. granarium F(3436) = 226.03 p = 0.000; C. ferrugineus F(3436) = 481.51 p = 0.000). Higher mortality was achieved with higher concentration that is maximum mortality against T. castaneum T. granarium and C. ferrugineus recorded at 20l/L concentration was 25.69 17.80 and 35.48 % respectively. Overall mean mortality of essential oils against all test insects at 5 l/L concentration was recorded 10.5% which was significantly increased to 26.72% with 20 l/L concentration. It is obvious from Table III regarding plant and concentration interaction that the highest fumigant toxicity (29.55 %) was depicted by T8 against T. castaneum and T4 (45.14%) against C. ferrugineus.

Fumigation with essential oils was performed for three different exposure times which significantly affected the mortality against T. castaneum (F(2436) = 814.41 p = 0.000) T. granarium (F(2436) = 572.21 p = 0.000) and C. ferrugineus (F(2436) = 728.55 p = 0.000). Mean mortality after 72hr (lowest exposure time) of treatment was 9.12 6.19 and 14.19% which significantly increased to 26.56 18.53 and 33.21% after 168hr (highest exposure time) against T. castaneum T. granarium and C. ferrugineus respectively. Essential oils and exposure time interaction significantly increased mortality of all test insects (Table I).

When the effects of exposure time for all the oils was checked the maximum mortality was observed in D. stramonium after 168hrs of treatment against C. ferrugineus (41.512.81) T. castaneum (33.831.72) and T. granarium (24.981.17) (T3 Table I).

All the results showed that the mean fumigant toxicity was concentration and exposure time dependent (Table II). Fumigation with higher concentration of essential oil for prolonged period of time (T12) resulted in 40.84 27.26 and 51.90 % mortality against T. castaneum T. granarium and C. ferrugineus respectively.

Table I.- Percent mortality of T. castaneum T. granarium and C. ferrugineus at different exposure periods against plant essential oils.

###Mortality (%)

Treatments

###T.###T.###C.

Exposed for (h)

###castaneum###granarium###ferrugineus

D. stramonium

###72###07.70.5 g###06.00.5 ge###14.60.8 f

###120###14.80.8 e###12.00.6 ef###29.01.6 c

###168###33.81.7 a###25.01.2 a###41.52.8 a

N. sativa

###72###10.30.7 fg###06.10.4 ge###10.00.5 g

###120###19.11.0 d###10.50.6 ef###23.61.5 de

###168###30.41.7 b###16.11.0 c###34.92.2 b

E. camaldulensis

###72###10.60.6 fg###07.50.5 g###16.10.7 f

###120###16.71.0 de###14.80.8 cd###21.81.0 de

###168###24.91.8 c###20.21.1 b###31.81.6 bc

M. oleifera

###72###07.80.5 g###05.00.5 e###16.00.8 f

###120###11.00.7 f###10.20.6 f###20.60.9 e

###168###17.01.1 de###12.80.9 de###24.61.2 d

Fumigant toxicity against T. castaneum

The essential oils significantly exhibited fumigant toxicity against T. castaneum. N. sativa was found the most lethal fumigant against T. castaneum with mean mortality of 20.06% followed by D. stramonium (18.90%) E. camaldulensis (17.43%) and M. oleifera (12.03%). Fumigation for longer time with higher concentration (T12 Table II) exhibited maximum mortality (40.84%). Three strains of T. castaneum investigated in this experiment significantly affected mortality (F(2436) = 16.233 p = 0.000). Susceptibility of TTS strain was found high (18.43% mortality) whereas moderate in SWL (16.93% mortality) and low susceptibility to essential oils was observed in FSD strain (15.96% mortality). Table IV regarding exposure time and strain interaction also indicated the same trend with maximum mortality in T9.

Table II.- Percent mortality of T. castaneum T. granarium and C. ferrugineus against different concentrations of all plant essential oils at three exposure times.

Exposed for###Mortality (%)

dose (l/L)###T. castaneum###T. granarium###C. ferrugineus

72 h

###5###5.1 0.37 i###3.1 0.4 h###8.8 0.5 h

###10###7.5 0.4 hi###4.9 0.4 gh###12.6 0.5 g

###15###10.2 0.5 gh###7.3 0.4 f###16.1 0.5 ef

###20###13.5 0.5 ef###9.4 0.4 ef###19.3 0.7 de

120 h

###5###9.3 0.5 h###7.2 0.4 fg###14.0 0.5 fg

###10###12.6 0.5 fg###10.5 0.5 e###19.8 0.5 d

###15###17.2 0.7 d###13.2 0.5 d###26.1 0.7 c

###20###22.7 0.8 c###16.7 0.6 c###35.3 1.1 b

168 h

###5###15.6 0.9 de###11.0 0.8 de###20.3 0.5 d

###10###22.2 1.0 c###16.5 0.8 c###27.3 0.7 c

###15###27.5 1.6 b###19.3 1.1 b###33.3 1.8 b

###20###40.8 1.5 a###27.3 0.9 a###51.9 2.1 a

Table III.- Percentage mortality of T. castaneum and C. ferrugineus against four concentration of plant essential oils.

Fumigant###Dose###Mortality (%)

###(l/L)###T. castaneum###C. ferrugineus

D. stramonium###5###12.7 1.4 fgh###16.11 1.2 jk

###10###15.7 1.7 def###22.41 1.3 gh

###15###19.7 2.1 bc###30.23 2.1 cd

###20###27.4 2.9 a###45.13 3.6 a

D. stramonium###5###11.8 1.0 ghi###12.52 1.0 k

###10###16.5 1.3 cde###18.58 1.5 hij

###15###22.3 1.9 b###25.01 2.1 ef

###20###29.5 2.2 a###35.62 3.0 b

E. camaldulensis###5###9.0 0.6 ij###15.32 0.9 jk

###10###14.3 1.0 efg###20.39 1.2 hi

###15###18.8 1.4 cd###24.85 1.4 ef

###20###27.6 1.9 a###32.63 1.9 bc

M. oleifera###5###6.6 0.5 j###13.49 0.7 k

###10###9.9 0.7 hij###18.12 0.7 ij

###15###13.4 1.0 efg###21.56 1.0 ghi

###20###18.2 1.2 cd###28.52 1.1 de

Though fumigant effect of essential oils was significant but T. granarium was found less susceptible with overall 12.27% mean mortality as compared to other test insects in this study. The higher mortality was exhibited by D. stramonium (14.46%) and E. camaldulensis (14.27%) followed by N. sativa (10.94%) and M. oleifera (9.41%). Effect of different strains on mortality was found significant (F(2436) = 17.269 p = 0.000). According to fumigant mortality in different strains following order was observed FSD (11.01%) less than SWL (12.62%) less than (13.12%) TTS. Essential oil and exposure time interaction was found significant which indicated maximum mortality (24.98%) in T3 against T. granarium (Table I).

Table IV.- Percentage mortality of three strains of T. castaneum and C. ferrugineus against essential oils at different exposure times.

Treatments time###Mortality (%)

###(h)###T. castaneum###C. ferrugineus

72 h

###FSD###8.4 0.5 d###13.5 0.7 d

###SWL###9.3 0.5 d###14.9 0.7 d

###TTS###9.7 0.5 d###14.1 0.7 d

120 h

###FSD###14.4 0.8 c###22.4 1.2 c

###SWL###15.5 0.8 c###24.2 1.1 c

###TTS###16.4 0.9 c###24.8 1.2 c

168 h

###FSD###25.1 1.5 b###31.6 1.7 b

###SWL###25.5 1.6 b###32.2 2.1 b

###TTS###29.2 1.7 a###35.8 1.8 a

Fumigant toxicity against C. ferrugineus

Strong fumigant insecticidal bioactivities of essential oils were recorded against C. ferrugineus as compared to T. granarium and T. castaneum. Essential oils depicted 23.79% overall mortality against C. ferrugineus while D. stramonium was found the most active fumigant (28.49%) followed by E. camaldulensis (23.31%) N. sativa (22.95%) and M. oleifera (20.44%). Effect of the strains on mortality was also found significant (F(2436) = 11.224 p = 0.00002) that is mean mortality of 22.52 23.97 and 24.89% was recorded in FSD SWL and TTS strain respectively. Significant interaction of strain and exposure time is given in Table IV which indicated maximum mortality (35.78%) in T9 against C. ferrugineus. Highest individual mean mortality of essential oils against C. ferrugineus was observed at 20% concentration after 168hr of treatment which was 70.46 55.93 45.76 and 35.44% against D. stramonium N. sativa E. camaldulensis and M. oleifera respectively.

DISCUSSION

This experiment was designed to evaluate fumigant potential of some indigenous medicinal plants. Effect of essential oils from D. stramonium E. camaldulensis M. oleifera and N. sativa was found significant against T. granarium C. ferrugineus and T. castaneum. Insecticidal activities of D. stramonium (Mahfuz and Khanam 2007) E. camaldulensis (Negahban and Moharramipour 2007) M. oleifera (Anita et al. 2012) and N. sativa (Chaubey 2007) are extensively investigated against stored grain insect pests.

In present studies D. stramonium (20.62%) was found the most effective fumigant followed by E. camaldulensis (18.34%) N. sativa (17.98%) and M. oleifera (13.96%). Whereas the highest mortality in T. castaneum (20.057 %) was recorded against N. sativa. These results are also in agreement with that of Chaubey (2007) who evaluated fumigant toxicity of Anethum graveolens Nigella sativa and Trachyspermum ammi against T. castaneum adults found N. sativa the most toxic among these essential oils. Among essential oils M. oleifera exhibited least mortality against T. castaneum (12.03%) T. granarium (9.38 %) and C. ferrugineus (20.44%).

T. granarium (12.25%) was found tolerant insect followed by T. castaneum (17.11 %) and C. ferrugineus (23.79%). These findings are similar to those of Ali et al. (2012) who evaluated insecticidal efficiencies of Datura alba and found T. granarium more tolerant than S. oryzae with 33.5 and 45% mortality respectively. In another study Abdel- Sattar et al. (2010) investigated insecticidal activities of Schinus molle against T. granarium and T. castaneum and found LC50s 915.1 and 779.1l/L which indicated higher tolerance in T. granarium than T. castaneum.

Fumigation with higher concentration of essential oils enhanced the mortality in insects. It is evident from Table III that treatment with higher concentrations T8 (29.55%) and T4 (45.13%) resulted in maximum mortality against T. castaneum and C. ferrugineus respectively. These findings are in accordance with earlier reports documented by Rozman et al. (2006) Negahban et al. (2006) Liska et al. (2010) and Theou et al. (2013).

Insects were fumigated with essential oils for 72 120 and 168 h and exposure time was found directly related to mortality. All investigated essential oils exhibited strongest fumigation action at 168 h of exposure to all tested insects. It is easily understandable from Table I that maximum mortality was observed in T3 against T. granarium C. ferrugineus and T. castaneum. These results are confirmatory to those of Ebadollahi et al. (2010) who evaluated fumigant toxicity of essential oil extracted from Lavandula stoechas against T. castaneum and observed decrease in LC50 by increasing exposure time i.e. LC50 was recorded 39.68 29.41 and 26.77l/L after 24 48 and 72 h respectively. Similar trend was observed by Liska et al. (2010) who recorded 5.0 % fumigant toxicity of camphor at 2 h and 13.2% after 4 h of treatment against T. castaneum.

These findings were also in accordance with previous studies carried out by Shukla et al. (2002) Lee et al. (2004) Rozman et al. (2006) and Xie et al. (2010).

Three strains of T. granarium C. ferrugineus and T. castaneum investigated in this experiment were found significantly different in susceptibility to essential oils. FSD strain was found tolerant as it showed less mortality (16.5%) against all essential oils followed by SWL (17.84%) and TTS (18.81%) strain. Results are quite in line to those of Khalequzzaman and Sultana (2006) who also reported the significant difference in sensitivity of different strains of T. castaneum against essential oils. Similar conclusions have also been drawn by Sagheer et al. (2013) who investigated Vehari and Faisalabad strains of T. granarium against some essential oils which resulted higher mortality against Vehari strain and proved Faisalabad (FSD) strain tolerant.

In summary the botanicals have a potential to control the stored grain pests at higher concentrations and for long periods of exposures. This study along with some other earlier studies opens a way for grain-storage protectionists to use natural oils in the huge storage structures instead of the synthetic fumigants to overcome the problem of insect resistance.

ACKNOWLEDGEMENTS

This study is supported by Higher Education Commission (HEC) Islamabad Pakistan (Batch-VI 106-2004-Av6-112) under the Indigenous Ph.D. Fellowship Program.

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Author:Saleem, Shahzad; Hasan, Mansoor ul; Sagheer, Muhammad; Sahi, Shahbaz Talib
Publication:Pakistan Journal of Zoology
Article Type:Report
Geographic Code:9PAKI
Date:Oct 31, 2014
Words:5240
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