Insecticidal Activity of Four Plant Essential Oils against Two Stored Product Beetles

Karim Saeidi; Hossein Pezhman

doi:10.4172/2161-0983.1000213

Research Article - (2018) Volume 7, Issue 3

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Insecticidal Activity of Four Plant Essential Oils against Two Stored Product Beetles

Karim Saeidi^* and Hossein Pezhman: Department of Plant Protection Research, Fars Agricultural and Natural Resources Research and Education Center, AREEO, Shiraz, Iran, E-mail: hossein.pezhman@yahoo.com

^*Corresponding Author: Karim Saeidi, Department of Plant Protection Research, Fars Agricultural and Natural Resources Research and Education Center, AREEO, Shiraz, Iran, Tel: +987413334821 Email:

Abstract

The insecticidal properties of essential oils prepared using water distillation method from aerial parts of Mentha piperita, Mentha pulegium, Zataria multiflora and Thymus daenensis were investigated on adult of Bruchus lentis and Callosobruchus maculatus. Essential oils were used in five concentrations and five replications. Mortality rate of insects were recorded after 3, 6, 24, 48 and 72 hours. Results indicated that the longer exposure of insect to essential oil and the higher concentration of essential oil, increased mortality of the two species in all treatments. There was significant difference among mortality effect of the essential oils. The LC₅₀ value of M. piperita after 24 hours for B. lentis and C. maculatus was 14.62 and 13.70 μl/l air respectively, while the values of LC₅₀ were 92.32 and 95.80 for M. pulegium, 58.43 and 99.94 for Z. multiflora and 63.97 and 65.55 μl/l air for T. daenensis, respectively. The least LT₅₀ recorded was in T. daenensis essential oil with 26.21 and 24.15 hours against B. lentis and C. maculatus, respectively.

Keywords: Fumigation toxicity; The seed lentil beetle; The cowpea weevil; Essential oil; Yasouj

Introduction

In some rural areas of Iran that use traditional storages, damage caused by stored product insects can be as high as 80% [1]. Fumigation plays a major role in insect pest elimination in stored products [2]. Chemical control of stored products ‘pests with current chemical pesticides may cause special problems on stored products [3].

These problems have highlighted the need for the development of new types of selective insect-control alternatives with fumigant action [4]. It is believed that essential oils have the advantage over conventional fumigants in terms of low mammalian toxicity, rapid degradation and local availability [5]. They do not leave residues toxic to the environment and have medicinal properties for humans [6]. These are the by-products of plant metabolism and are commonly referred to as volatile plant secondary metabolites [7]. Because of the intensity of plant-insect interactions, the plants have well developed defense mechanisms against pests and are excellent sources of new insecticidal substances. Their components and quality vary with geographical distribution, time of harvest, growing conditions and method of extraction [8]. Among others effects of essential oils on stored-product insect pests have been reported on extensively [9-14].

Nowadays, many studies have been done for evaluation of susceptibility of stored product insect pests specially Callosobruchus maculatus to plant essential oils. For example, the antifeedant activities of Citrus reticulate Blanco, Citrus limon L. and Citrus aurantium L. essential oils against eggs, larvae and adults of C. maculatus were studied [15]. The results showed that the effect of different concentrations of the essential oil vapors on egg hatchability as well as larval and adult mortality was found to be significant. C. reticulata and C. aurantium oils were more toxic on egg hatchability than C. limon extract and caused higher mortality on larvae as well. In the study of [15], the essential oils of Eucalyptus globulus and Eucalyptus camaldulensis against C. maculatus was evaluated. The results revealed that E. globulus oils were more effective than E. camaldulensis oils, by significantly decreasing the RGR, RCR and ECI. Both of plant essential oils, with the same activity, increased FDI as the oil concentration was increased, showing high feeding deterrence activity against C. maculatus.

In the present study, also for importance of insecticidal activity in the integrated pest management, this effect was investigated by essential oils of M. piperita, M. pulegium, Z. multiflora and T. daenensis grown in Iran against adults of B. lentis and C. maculatus two important storage pests.

Materials and Methods

Plant materials

The aerial parts (stem and leaves) M. piperita, M. pulegium, Z. multiflora and T. daenensis were collected at the ripening stage in the Dena protected area, Sisakht city, of the province Kohgiluyeh va Boyerahmad Ahmad (Iran), in July 2015. Plant taxonomists in the Department of Biology at Urmia University (Iran) confirmed the taxonomic identification of the plant species. The voucher specimens have been deposited at the herbarium of the Department of Natural Resources in, Kohgiluyeh va Boyerahmad Agricultural and Natural Resources Research and Education Center, Yasouj, Iran.

Insect rearing

Rearing of Bruchus lentis: The culture medium was the whole lentil grains sterilized at 60°C for 60‐90 minutes. Ten jars of 300 mg were used. Each jar was filled with 250 g lentil grains and 30 beetles were added to each jar. The jars were then covered with muslin cloth, tied with rubber bands to avoid the escape of beetles. Beetles were left in the culture medium for 3 days for egg laying and then were removed with the help of sieves and fine camel hair brushes. The lentil grains containing eggs were placed again in the same jars and put in the incubator for incubation at 30 ± 2°C and 65% RH to get the homogenous (same age) population. Relative humidity was maintained inside the incubator by placing an open tray filled with saturated solution of NaNO₂.

Rearing of Callosobruchus maculatus: A culture of the C. maculatus was established on the seeds of unshelled brown cowpea, Vigna ungiculata L. in one litter wide-mouthed glass jars under laboratory conditions. Parent adults were obtained from laboratory stock culture maintained at the Entomology Department, University of Urmia, Iran. The culture was maintained in the dark (similar to storage conditions) in a growth chamber set at 27 ± 1°C and 65 ± 5 % relative humidity.

All experiments were carried out under the same environmental conditions. The 1-2 days old adults of C. maculatus were used in bioassay tests.

Essential oil supply

Plant materials of M. piperita, M. pulegium, Z. multiflora and T. daenensis were air dried in the shade at room temperature (26-28°C) for 20 days and stored in darkness until distillation. The essential oils were isolated from dried plant samples by hydro-distillation using a Clevenger apparatus. Conditions of extraction were: 50 g of air-dried sample, 1:10 plant material/water volume ratio, 3 hours distillation. The essential oils were dried over anhydrous sodium sulfate and stored in glass tubes at +4°C in refrigerator.

Bioassay

Fumigant toxicity of essential oils: In fumigant toxicity assays, filter papers (What man No. 1, cut into 4 × 5 cm paper strip) were impregnated with different concentrations using a micro sampler. Twenty adults (3-5 days old) of B. lentis and C. maculatus were placed in small plastic tubes (3.5 cm diameter and 5 cm height) with open ends covered with cloth mesh. The tubes were hung at the geometrical center of glass bottles, which were then sealed with air–tight lids. The experimental concentrations of essential oils on B. lentis and C. maculatus adults were determined by preliminary tests. Experimental concentrations for M. piperita were 10, 11, 12, 13 and 14 μl/l air of oil and for Z. multiflora and T. daenensis were 40, 50, 60, 70 and 80 μl/l air of oils and for M. pulegium were 60, 70, 80, 90 and 100 μl/l air of oil. In the control bottles, only acetone was applied on the filter papers. Each experiment was replicated five times for each concentration. In all cases, the exposure times were 3, 6, 24, 48 and 72 hours. Treated insects were incubated at 27 ± 1°C. After this time, the number of dead adults was counted. Those adults that did not move when lightly probed or shaken in light and mild heat were considered dead. Mortality in the control was not observed in any experiment.

Statistical analysis

The mortality data were corrected using Abbott’s formula [16] for the mortalities in the controls, and then subjected to probit analyses using SPSS (version 16.0) software to estimate LC₅₀ value. The percentage mortality value for different exposure times were subjected to analysis of variance (one-way ANOVA) using the SPSS (version 16.0) software statistical program. Data were transformed using arcsine √x to meet normality, before ANOVA. Comparison of means was done through Tukey’s (HSD) test at 0.05.

Results

Among investigated essential oils, M. piperita essential oil has more fumigant toxicity than others on both species. With increasing concentrations of M. piperita essential oil, the mortality of mentioned species was increased so that the highest concentration (14 μl/l air) caused 100% mortality on both adult insects after 72 hours (F=3.01 P < 0.01) (Table 1), but T. daenensis essential oil in 80 μl/l air concentration caused 100% mortality on both adult insects (F=2.23 P<0.01), (Table 2). The essential oil of Z. multiflora in 80 μl/l air concentration caused 100 and 83% mortality respectively on both adult insects after 72 hours (F=42.51 P< 0.01), (Table 3). M. pulegium essential oil in 100 μl/l air concentration caused 85 and 89% mortality respectively on both adult insects after 72 hours and had the lowest insecticidal effect in comparison to other essential oils (F = 6.92 P < 0.01), (Table 4). Calculated LC₅₀ indicated that the LC₅₀ of M. piperita essential oil on B. lentis and C. maculatus was 14.62 μl/l and 13.70 μl/l air concentrations respectively while the LC₅₀ of T. daenensis essential oil for these insects was 63.97 and 65.55 μl/l air concentration respectively. The LC₅₀ of oil was 58.43 and 99.94 μl/l air concentration respectively.

Means of mortality% ± S. E
Time (Hour)
Insect	Con(μl/lair)	3	6	24	48	72
B. lenis	10	0.00 ± 0.00ⁿ	0.00 ± 0.00ⁿ	27 ± 2.53^ml	37 ± 2.32^jkl	59 ± 3.00^efg
	11	0.00 ± 0.00ⁿ	0.00 ± 0.00ⁿ	27 ± 2.51^ml	43 ± 3.20^hijk	75 ± 4.23^bcd
	12	0.00 ± 0.00ⁿ	0.00 ± 0.00ⁿ	31 ± 3.69^jklm	43 ± 6.04^hijk	89 ± 6.20^ab
	13	0.00 ± 0.00ⁿ	0.00 ± 0.00ⁿ	41 ± 11.25^hijkl	45 ± 7.00^jhig	89 ± 5.00^ab
	14	0.00 ± 0.00ⁿ	0.00 ± 0.00ⁿ	55 ± 3.00^fgh	71 ± 4.45^cde	100 ± 0.00^a
C. maculatus	10	0.00 ± 0.00ⁿ	0.00 ± 0.00ⁿ	21 ± 5.10^m	31 ± 5.42^jklm	45 ± 4.22^ghij
	11	0.00 ± 0.00ⁿ	0.00 ± 0.00ⁿ	29 ± 3.20^klm	39 ± 4.25^ijkl	65 ± 2.17^def
	12	0.00 ± 0.00ⁿ	0.00 ± 0.00ⁿ	35 ± 6.13^jklm	53 ± 6.56^fghi	85 ± 3.00^bc
	13	0.00 ± 0.00ⁿ	0.00 ± 0.00ⁿ	53 ± 4.21^fghi	67 ± 3.00^def	89 ± 2.27^ab
	14	0.00 ± 0.00ⁿ	0.00 ± 0.00ⁿ	53 ± 5.21^fghi	77 ± 3.14^bcd	100 ± 0.00^a

*Mean in the same column followed by the same letters are not significantly different as determined by the Tukey’s-test.

Table 1: Average mortality caused by Mentha piperita essential oil on B. lentis and C. maculatus at concentrations and different times.

Means of mortality% ± S. E
Time (Hour)
Insect	Con(μl/l air)	3	6	24	48	72
B. lenis	40	0.00 ± 0.001	0.00 ± 0.001	35 ± 2.20^hijk	39 ± 4.16^ghij	45 ± 7.00^fghi
	50	0.00 ± 0.001	0.00 ± 0.001	35 ± 2.20^hijk	41 ± 7.21^ghij	55 ± 3.25^defg
	60	0.00 ± 0.001	0.00 ± 0.001	35 ± 2.12^hijk	53 ± 3.25^efgh	73 ± 4.18^bcd
	70	0.00 ± 0.001	0.00 ± 0.001	55 ± 11.32^defg	83 ± 4.20^bc	87 ± 5.00^abc
	80	0.00 ± 0.001	0.00 ± 0.001	63 ± 4.53^def	85 ± 5.52^abc	100 ± 0.00^a
C. maculatus	40	0.00 ± 0.001	0.00 ± 0.001	21 ± 3.60^m	39 ± 3.00^ghij	47 ± 6.20^fghi
	50	0.00 ± 0.001	0.00 ± 0.001	29 ± 3.12^klm	45 ± 4.12^fghi	55 ± 2.41^defg
	60	0.00 ± 0.001	0.00 ± 0.001	35 ± 4.41^jklm	47 ± 6.23^fghi	69 ± 3.15^cde
	70	0.00 ± 0.001	0.00 ± 0.001	53 ± 2.24^fghi	81 ± 5.34^bc	87 ± 4.20^abc
	80	0.00 ± 0.001	0.00 ± 0.001	53 ± 3.28^fghi	91 ± 6.25^ab	100 ± 0.00^a

*Mean in the same column followed by the same letters are not significantly different as determined by the Tukey’s-test.

Table 2: Average mortality caused by Thymus daenensis essential oil on B. lentis and C. maculatus at concentrations and different times.

Means of mortality% ± S. E
Time (Hour)
Insect	Con (μl/l air)	3	6	24	48	72
B. lenis	40	0.00 ± 0.00^k	0.00 ± 0.00^k	27 ± 9.27^ij	41 ± 9.27^fghi	59 ± 4.00^cdef
	50	0.00 ± 0.00^k	0.00 ± 0.00^k	41 ± 3.74^fghi	53 ± 6.32^defg	61 ± 2.15^cde
	60	0.00 ± 0.00^k	0.00 ± 0.00^k	47 ± 8.72^efg	69 ± 5.10^bcd	81 ± 4.45^ab
	70	0.00 ± 0.00^k	0.00 ± 0.00^k	57 ± 8.12^cdef	69 ± 8.72^bcd	89 ± 3.12^a
	80	0.00 ± 0.00^k	0.00 ± 0.00^k	63 ± 5.48^defg	71 ± 5.83^bc	100 ± 5.07^a
C. maculatus	40	0.00 ± 0.00^k	0.00 ± 0.00^k	21 ± 3.74^jk	35 ± 3.74^ghij	51 ± 3.00^fg
	50	0.00 ± 0.00^k	0.00 ± 0.00^k	23 ± 4.47^j	41 ± 4.90^fghi	53 ± 2.34^cdefg
	60	0.00 ± 0.00^k	0.00 ± 0.00^k	29 ± 2.45^hij	49 ± 4.00^efg	69 ± 3.23^bcd
	70	0.00 ± 0.00^k	0.00 ± 0.00^k	35 ± 5.83^ghij	51 ± 3.74^defg	71 ± 4.22^bc
	80	0.00 ± 0.00^k	0.00 ± 0.00^k	45 ± 3.74^efgh	57 ± 5.10^cdef	83 ± 5.11^ab

*Mean in the same column followed by the same letters are not significantly different as determined by the Tukey’s-test.

Table 3: Average mortality caused by Zataria multiflora essential oil on B. lentis and C. maculatus at concentrations and different times.

Means of mortality% ± S. E
Time (Hour)
insect	Con(μl/lir)	3	6	24	48	72
B. lenis	60	0.00 ± 0.00^j	0.00 ± 0.00^j	33 ± 0.00^hi	39 ± 3.32^ghi	47 ± 3.00^fgh
	70	0.00 ± 0.00^j	0.00 ± 0.00^j	39 ± 4.20^ghi	45 ± 2.14^fgh	59 ± 3.13^cdef
	80	0.00 ± 0.00^j	0.00 ± 0.00^j	45 ± 2.54^fgh	55 ± 4.17^fgh	67 ± 3.00^cde
	90	0.00 ± 0.00^j	0.00 ± 0.00^j	53 ± 6.21^efg	71 ± 4.12^abcde	75 ± 3.10^abc
	100	0.00 ± 0.00^j	0.00 ± 0.00^j	57 ± 3.12^cdefg	71 ± 6.23^abcde	85 ± 2.15^ab
C. aculatus	60	0.00 ± 0.00^j	0.00 ± 0.00^j	21 ± 2.34^ij	33 ± 4.15^hi	41 ± 5.17^fgh
	70	0.00 ± 0.00^j	0.00 ± 0.00^j	29 ± 5.14^hi	41 ± 5.11^fgh	45 ± 3.25^fgh
	80	0.00 ± 0.00^j	0.00 ± 0.00^j	29 ± 3.20^hi	43 ± 11.17^fgh	57 ± 7.12^cdefg
	90	0.00 ± 0.00^j	0.00 ± 0.00^j	53 ± 6.04^efg	69 ± 4.23^bcde	75 ± 6.15^abc
	100	0.00 ± 0.00^j	0.00 ± 0.00^j	59 ± 4.00^cdef	73 ± 7.31^abcd	89 ± 3.23^a

*Mean in the same column followed by the same letters are not significantly different as determined by the Tukey’s-test.

Table 4: Average mortality caused by Mentha pulegium essential oil on B. lentis and C. maculatus at concentrations and different times.

The highest calculated LC₅₀ belonged to M. pulegium essential oil with 92.32 and 95.80 μl/l air on B. lentis and C. maculatus, respectively (Table 5). Based on LC₅₀ and lower and upper confidence interval 95% the LC₅₀ of M. piperita essential oil on B. lentis and C. maculatus with 14.62 and 13.70 μl/l air had significantly different with other essential oils. The findings indicated that there wasn’t any significantly different between the effects of T. daenensis and M. pulegium essential oils based on LC₅₀ of the adults of B. lentis. But the essential oil of Z. multiflora was significantly different with T. daenensis and M. pulegium essential oils. Also, there wasn’t any significantly different between effects of M. pulegium, Z. multiflora and T. daenensis essential oils on C. maculatus and they had overlap based on LC₅₀ and confidence interval 95%. There wasn’t any significantly difference between the effects of all studied essential oils on mortality of both B. lentis and C. maculatus adults (Tables 1-4).

Essenial oil	Insect	N	X²(df = 3)	Slope ± S. E	LC₅₀ (μl/lair)	95% (Confidence limit)
Essenial oil	Insect	N	X²(df = 3)	Slope ± S. E	LC₅₀ (μl/lair)	Lower	Upper
Mentha piperita	B. lentis	300	2.12	6.92 ± 1.54	14.62	12.25	19.75
Mentha piperita	C. maculatus	300	1.05	8.09 ± 1.56	13.70	11.82	14.82
Mentha pulegium	B. lentis	300	0.01	5.09 ± 0.96	92.32	76.75	161.16
Mentha pulegium	C. maculatus	300	1.30	6.69 ± 0.99	95.80	84.81	116.30
Zataria multiflora	B. lentis	300	0.16	5.58 ± 0.96	58.43	49.99	71.54
Zataria multiflora	C. maculatus	300	0.78	4.98 ± 0.99	99.94	73.75	442.75
Thymus daenensis	B. lentis	300	2.40	5.65 ± 0.97	63.97	57.50	82.96
Thymus daenensis	C. maculatus	300	4.06	6.77 ± 0.99	65.55	59.83	78.23

Table 5: Calculated values LC₅₀ of essential oils from M. piperita, M. pulegium, Z. multiflora and T. daenensis on B. lentis and C. maculatus after 24 h.

The results showed that in 3 and 6 hours after applying of all concentrations of essential oils the mortality of both adult insects was zero and mortality was started for 24 hours after applying essential oils and reached after 72 hours to maximum after applying essential oils. In all studied essential oils, there was significantly difference between mortalities occurred for each concentration in 24, 48 and 72 hours after applying essential oils. Calculated LT₅₀ for essential oils revealed that T. daenensis essential oil had the least LT₅₀ for both species so that it was 26.21 and 24.15 hours for B. lentis and C. maculatus respectively. The LT₅₀ of M. piperita essential oil on B. lentis and C. maculatus was 29.66 and 29.43 hours respectively. The LT₅₀ of Z. multiflora essential oil on B. lentis and C. maculatus was 28.45 and 39.62 hours respectively and the LT₅₀ of M. pulegium essential oil on B. lentis and C. maculatus was 33.27 and 30.32 hours was calculated respectively (Table 6).

Essenial oil	Insect	N	X²(df = 3)	Slope ± S. E	LT₅₀ (Hour)	95% (Confidence limit)
Essenial oil	Insect	N	X²(df = 3)	Slope ± S. E	LT₅₀ (Hour)	Lower	Upper
Mentha piperita	B. lentis	200	12.02	5.67 ± 0.65	29.66	8.60	50.17
Mentha piperita	C. maculatus	200	7.65	5.94 ± 0.55	29.43	15.42	42.63
Mentha pulegium	B. lentis	200	7.48	4.62 ± 0.35	33.27	19.66	54.98
Mentha pulegium	C. maculatus	200	6.11	4.88 ± 0.23	30.32	26.92	36.84
Zataria multiflora	B. lentis	200	9.99	5.13 ± 0.26	28.45	13.43	47.53
Zataria multiflora	C. maculatus	200	5.24	4.71 ± 0.14	39.62	34.49	47.45
Thymus daenensis	B. lentis	200	6.07	5.99 ± 0.23	26.21	22.21	29.89
Thymus daenensis	C. maculatus	200	4.41	6.24 ± 0.33	24.15	21.36	28.62

Table 6: Calculated values LT₅₀ of essential oils from M. piperita, M. pulegium, Z. multiflora and T. daenensis on B. lentis and C. maculatus.

Discussion

According to the results with increasing in essential oil concentration, the adult mortality rate of both species was increased, which are in agreement with reports of other researcher [7,4,12,17], but in a similar experiment conducted by [18], the B. lentis sensitivity to Achillea wilhelmsii and Allium sativum essential oil is less than T. daenensis essential oil, which is evaluated in present study, so that, the LC₅₀ of A. wilhelmsii essential oil on this insect 122.1 μl/l air was determined while in this study, LC₅₀ of essential oil on mentioned insect 63.97 μl/l air was evaluated.

According to several surveys, the main active ingredients of M. piperita essential oil are menthone and menthol [19] and thymol and 1, 8-ciniole were reported as the main active ingredients of T. daenensis [20].

Based on the present results with increasing the concentrations of essential oils, the mortality of adults of both species was increased, that these results are in agreement with other similar reports [21]. Studies carried out by [22] showed that the sensitivity of Tribolium castaneum to Artemisia aucheri essential oil less than Artemisia hausskenechtii essential oil. The LC₅₀ of A. aucheri essential oil was reported 122.1 μl/l air by [22], while in present study; the LC₅₀ of T. daenensis essential oil on this mentioned species was 63.97 μl/l airs. More researchers stated that the major insecticidal properties of T. daenensis essential oil related to Thymol ingredient and it seems that the more toxicity of M. piperita on B. lentis in the present study is related to menthone. In another study A. wilhelmsii essential oil at a concentration of 1 μl/l caused 80% mortality of adult females of Tribolium confusum 48 hours after application [17], but in the present work, the concentration of 80 μl/l of Z. multiflora caused such a mortality for mentioned insect. This different result may be due to differences in plant active ingredient so that there are reports that show the active ingredient of a plant species is varied in terms of quantitative and qualitative in different geographical conditions [23].

Reference [17] showed that the LC₅₀ of M. piperita essential oil on T. castaneum was 573.94 μl/l air that in comparison to T. daenesis essential oil that was evaluated in this study had lower fumigant toxicity on B. lentis. Also, [17] Calmasur et al. reported the LC₅₀ of Mentha longifolia essential oil on T. castaneum 0.56 μl/l air, that according to this results the toxicity of essential oils studied are more than our results in the present study [24-26].

This difference may be related to more thymol in Thymus than other species of this genus thymol was demonstrated as an effective insecticide by several researchers. The results of present study revealed that among four essential oils that were surveyed on two species, M. piperita essential oil had high fumigant toxicity for both species and due to the low risk of essential ingredients of this plant for human health and the environment can be used in pest management programs.

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Citation: Karim S, Hossein P (2018) Insecticidal Activity of Four Plant Essential Oils against Two Stored Product Beetles. Entomol Ornithol Herpetol 7: 213.

Copyright: Â© 2018 Karim S, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Entomology, Ornithology & Herpetology: Current ResearchOpen Access

Insecticidal Activity of Four Plant Essential Oils against Two Stored Product Beetles

Abstract

Introduction

Materials and Methods

Results

Discussion

References

Entomology, Ornithology & Herpetology: Current Research
Open Access