• No results found

Evaluation of Origanum vulgare L. essential oil as a source of toxicant and an inhibitor of physiological parameters in diamondback moth, Plutella xylustella L. (Lepidoptera: Pyralidae)


Academic year: 2021

Share "Evaluation of Origanum vulgare L. essential oil as a source of toxicant and an inhibitor of physiological parameters in diamondback moth, Plutella xylustella L. (Lepidoptera: Pyralidae)"


Loading.... (view fulltext now)

Full text



Evaluation of Origanum vulgare L. essential oil as a

source of toxicant and an inhibitor of physiological

parameters in diamondback moth, Plutella

xylustella L. (Lepidoptera: Pyralidae)

Mahsa Nasr


, Jalal Jalali Sendi


, Saeid Moharramipour


, Arash Zibaee



Department of Plant Protection, Faculty of Agricultural Sciences, University of Guilan, Rasht, Iran


Department of Entomology, Tarbiat Modares University, Tehran, Iran

Received 24 December 2013; revised 17 April 2015; accepted 30 June 2015

KEYWORDS Diamond back moth; Origanum vulgare; Toxicity; Repellency; Feeding efficiency; Biochemical effects

Abstract The effect of essential oil of Origanum vulgare L. a medicinal plant was studied on toxicity, physiology and biochemical characteristics of diamondback moth Plutella xylustella L. in controlled condition. The LC10, LC30 and LC50values were estimated 0.234, 0.710 and

1.528 percent (v/v) respectively for 3rd instar larvae. Repellency of essential oil under LC10 and LC30 concentrations was 22.8 ± 6.64 and 49.8 ± 6.95 percent respectively. Effect of plant essential oil on feeding efficiency was also evaluated under LC10and LC30concentrations. Approximately

Digestibility (AD), Efficacy of Conversion of Ingested Food (ECI), Efficacy of Conversion of Digested Food (ECD), Relative Consumption Rate (RCR) and Relative Growth Rate (RGR) of the treated larvae showed a significant difference compared with the control. The effectiveness of plant essential oil on digestive enzymes under LC50 concentration for the survivors after 24 and 48 h after treatment was analyzed. Total protein and triglycerides were decreased significantly com-pared with the control. Activity of alkaline phosphatase and protease in treated larvae decreased compared with the controls. The lipase was increased 24 h after treatment compared with the con-trol. Significant differences in detoxifying enzymes such as glutathione S-transferase and esterase were noted compared to controls. It is concluded that the essential oil used in the present experi-ment shows toxicity in higher doses. It is also concluded that the essential oil used did show

* Corresponding author. Fax: +98 1316690281. E-mail address:jjalali@guilan.ac.ir(J. Jalali Sendi). Peer review under responsibility of King Saud University.

Production and hosting by Elsevier

King Saud University

Journal of the Saudi Society of Agricultural Sciences



considerable changes in repellency, feeding efficiency, reduced larval weight and changes in bio-chemical properties which may nominate it for further investigation in key insect pests.

ª 2015 The Authors. Production and hosting by Elsevier B.V. on behalf of King Saud University. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

1. Introduction

The diamondback moth (DBM), Plutella xylostella, is a major and cosmopolitan pest of crucifer crops (Talekar and Shelton, 1993). This pest is present wherever its host plants exist and is considered to be the most widely distributed of all Lepidoptera (Shelton, 2004). Outbreaks of DBM in Southeast Asia some-times cause crop losses of more than 90% (Verkerk and Wright, 1996). Second, third and fourth instar larvae are sur-face feeders and they consume leaves, buds, flowers, the green outer layer of stems and also developing seeds (Anonymous, 1996). The DBM was the first crop insect to be reported resis-tant to DDT in 1953 in Java, Indonesia (Ankersmit, 1953), P. xylostellahas a high tendency to develop resistance to insec-ticides because of its high reproductive capability (Shelton et al., 1983) and now in many crucifer producing regions it has shown significant resistance to almost every insecticide applied in the field including new chemicals such as spinosyns, avermectins, neonicotinoids, pyrazoles and oxadiazines (Sarfraz and Keddie, 2005; Sarfraz et al., 2005).

In the past, control of P. xylostella relied heavily on the use of synthetic chemical insecticides, which has resulted in the development of resistance to all modern synthetic insecticides that have been used intensively for any prolonged length of time (Talekar and Shelton, 1993). However, in other parts of the world P. xylostella has already developed resistance to Bacillus thuringiensis Berliner products (Tabashnik, 1994). Therefore the development of alternative control methods is essential if this pest is to be managed successfully.

Plant essential oils have been suggested as alternative sources for insect control products because some are selective, biodegrade to nontoxic products, and have lower effect on non target organisms and the environment (Singh and Upadhyay, 1993; Isman, 2000, 2001). Many plant essential oils show acute toxicity, developmental disruption, repellency, and feeding deterrence against many insect species due to their complex mixtures of monoterpenoids and related phenols (Singh and Upadhyay, 1993; Wink, 1993; Isman, 2000; Shekari et al., 2008; Lingathurai et al., 2011; Neto Bandeira et al., 2013; Yazdani et al., 2013) This study aimed at assessing the potential of Origanum essential oil for its possible effect on P. xylostella that may serve for future control of this insect based on environment friendly substances.

2. Materials and methods 2.1. Essential oil preparation

The plant O. vulgare was collected from Maranta flower grow-ers, Ramsar north of Iran. The collected aerial parts were washed with water and then dried in the shade. The dried sam-ples were first made into powder and were then subjected to hydro distillation for 2 h using a Clevenger type apparatus.

After distillation, the oil obtained was isolated from water and dried over anhydrous Na2SO4.

2.2. Insect rearing

For the study, larvae of different ages were collected from the field and reared under laboratory condition (24 ±C, 75 ± 5% RH and 16:8 h LD photoperiods) in rearing jars (5d· 10 h) cm. The larvae were provided with fresh canola leaves (Opera var.) for feeding. After raising a generation the third instar larvae of next generation of cohort age were used for bioassays. The highest and lowest effective concentration was determined using logarithmic distance, and the intermediate concentrations were selected. The ultimate tests to determine the LC50, 4 replicates of 10 larvae were used after 24 h. The essential

oil was dissolved in methanol and rapeseed leaf disks, each with a diameter of 6 cm were immersed in the solution for 10 s, air dried and then placed in Petri dishes. In all experiments, the con-trol group was treated with methanol alone.

2.3. The effect of essential oil on Nutrition

To evaluate the effect of different oils on feeding, sublethal concentrations of LC30and LC10were used. All weights were

measured using a monopan balance accurate to 0.1 mg. In this experiment 4 replicates of 10 newly hatched larvae in each replicate were used. The larvae were starved for 4 h in order to empty their stomach contents. The leaf disks with a diame-ter of 8 cm was weighed and then immersed for 20 s in desired concentration, air dried and then provided to the larvae. The larvae were weighed and then they were allowed to feed on the treated or control leaves. After 24 h the remaining leaves were removed and replaced with fresh treated leaves. The feces generated at the end of each day were collected, oven dried and then weighed. Larval weight was recorded at the end of the experiment. At the end of the experiment, some larvae were oven dried and weighed again to determine the dry weight of larvae. This experiment lasted for 3 days and observations were recorded at the end of each day.

To determine nutritional indices the following formulae by (Waldbauer, 1968) were used:

Approximate digestibility (AD) = 100 (E F)/F,

Efficiency of conversion of ingested food (ECI) = 100P/E,

Efficiency of digested food (ECD) = 100 P/(E F),

Consumption index (CI) = E/TA and Relative growth rate (RGR) = P/TA.


A= average of dry weight of larvae during the experiment. E= dry weight of consumed food.

F= dry weight of produced feces. P= dry weight of the biomass of larvae. T= duration of the experiment (3 days).


2.4. Repellent activity

To examine the effect of repellency of essential oils, the method of (Smith et al., 1994), was used. For this purpose, a hole was provisioned on each side of a cubic shaped plastic container (65 mL capacity). A 2 cm long· 3 mm diameter plastic tube connected each side of the container to two similar containers. The larvae could freely move through tube. To determine repellence activity of the essential oil, two sublethal concentra-tions of LC10and LC30based on previous toxicity tests were

selected. A control leaf disk of canola (6 cm) was placed in the left container impregnated with methanol and dried at room temperature. A treated leaf with either LC10 or LC30

and dried at room temperature was placed in the left container. This experiment was replicated 5 times with 10 third instar larvae of diamond back moth in each replication that were released in the center of middle container. After 24 h the number of larvae in each container was counted. The following formula ofLiu et al. (2006)was used for calculation: Repellencyð%Þ ¼ ðC  EÞ=Tð Þ  100

where C is the number of insects in control leaf, E is the number of insects in essential oil treated leaf and T is the total number of insects.

2.5. Biochemical tests

The remaining larvae after LC50 treatment were used after

24 and 48 h for various biochemical tests. 2.5.1. Measurement of total protein

Protein concentrations were determined by the method of (Bradford, 1976). First, each larva (whole body) was homoge-nized in 350 lL of distilled water and samples were centrifuged at 10,000g for 5 min at 4C. Then 10 lL of supernatant was mixed with 90 lL of distilled water and 2500 lL dye (10 mg powder of Coomassie Brilliant Blue (Bio-Rad, Munchen, Germany) in 5 ml 96% ethanol (96%) and 10 mL phosphoric acid 85% (w/w) then solution brought to 100 mL with distilled water). The absorbance was read at 630 nm.

2.5.2. Triglyceride measurements

A diagnostic kit from PARS-AZMOON Co. was used to measure the amount of triacylglyceride in the fourth instar lar-vae. Reagent solution contained phosphate buffer (50 mM, pH

7.2), 4-chlorophenol (4 mM), Adenosine Triphosphate

(2 mM), Mg2+(15 mM), glycerokinase 0.4 kU/L), peroxidase (2 kU/L), lipoprotein lipase (2 kU/L), 4-aminoantipyrine (0.5 mM) and glycerol-3-phosphate-oxidase (0.5 kU/L). Samples (10 ll) were incubated with 10 ll of distilled water and 70 ll of reagent for 20 min at 25C (Fossati and Prencipe, 1982). ODs of samples and reagent as standard were read at 546 nm.

2.5.3. Measurement of amylase activity

The a-amylase activity was measured by the procedure of (Bernfeld, 1955), using 1% soluble starch as substrate. The reaction was performed by sodium phosphate buffer at 35C with 10 lL of the enzyme, 40 lL substrate and 40 lL sodium phosphate buffer (pH = 9) for 30 min. To stop the reaction,

100 lL dinitrosalicylic acid (DNS) was added and heated in boiling water for 10 min. Absorbance was read at 540 nm after cooling. One unit of a-amylase activity was defined as the amount of enzyme required to produce 1 mg maltose in 30 min at 35C. In this test, maltose was used to produce stan-dard curve.

2.5.4. Measurement of protease activity

Protease activity was determined using azocasein as substrate (Garcia-Carreno and Haard, 1993). Each larva was cen-trifuged in 10 lL distilled water, then 10 lL of supernatant and 15 lL buffer (pH 7) with 50 lL of 1% azocasein were reacted for 3 h at 37C. Proteolysis was stopped by the addition of 150 lL of 10% trichloroacetic acid (TCA). The solution was transferred to 4C in a refrigerator for 30 min, and the reaction mixture was centrifuged at 12,000g for 10 min. Hundred lL of supernatant was mixed with 100 lL 1 N NaOH and the absorbance was read at 440 nm.

2.5.5. Measurement of lipase activity

The activity of lipase was determined using the method of (Tsujita et al., 1989). Ten lL of homogenate was mixed with 18 lL P-nitrophenyl Butyrate as substrate and mixed with 172 lL buffer (pH 7). The absorbance was read at 405 nm. 2.5.6. Measurement of alkaline phosphatase activity

Alkaline phosphatase activity was measured by the method of (Bessey et al., 1946). Fifty lL phosphate buffer (pH 8) was mixed with 30 lL of the enzyme substrate p-nitrophenolate and 15 lL of sample was added to the mixture and then activ-ity was read at 405 nm.

2.5.7. Esterase activity

The activity of general esterases was determined according to the method of (Van Asperen, 1962). a-naphtlyacetate (a-NA) and b-naphtylacetate (b-NA) (10 mM) were used as substrates. Initially one insect was homogenized with 1000 ml 0.1 M phos-phate (pH 7) containing Triton X-100 in the ratio of 0.01%, and centrifuged at 10,000g for 10 min at 48C. The super-natant was transferred to new micro tube and was diluted with phosphate buffer. Fast Blue RRsalt (1 mM) was added and the absorbance was read at 630 nm.

2.5.8. Glutathione S-transferase activity

For determining glutathione S-transferase (GST) activity, the method of (Habing et al., 1974) was used. 1-chloro-2, 4-dinitrobenzene (CDNB) (20 mM) was used as the substrate. Each larva was homogenized in 20 lL distilled water and centrifuged at 12,500g for 10 min at 48C. Fifteen lL of super-natant was mixed with 135 lL of phosphate buffer (pH = 7), 50 lL of CDNB and 100 lL of GST. The absorbance was read at 340 nm.

2.5.9. Statistical analysis

For determination of mortality and lethal concentration, POLO-PC software (LeOra, 1987) was used. The data from other experiments were subjected to analysis of variance (ANOVA) using SAS software. The least significant among treatments was compared using Tukey’s multiple range test


(SAS Institute, 1997). Differences between the various treat-ments were determined at 5% by Tukey’s multiple.

3. Results

Acute toxicity of Origanum essential oil on third instar larvae of diamond back moth is shown in (Table 1). In this experi-ment, increasing concentrations of herbal essential oil increased the mortality of larvae.

Repellent activity for essential oil after 24 h is shown in (Table 2). With increasing essential oil concentrations, the per-centage of repellency showed an increase and in highest and lowest concentrations it was 49.8 ± 6.95% in LC30(0.710%)

and 22.8 ± 6.64% in LC10 (0.234%) respectively (F = 5.42,

df= 2, p < 0.0484).

The results of the effect of sublethal concentrations of LC10

(0.234%) and LC30 (0.710) essential oil on feeding index are

shown in (Table 3). ECI (F = 8.46, df = 2, p < 0.0086), ECD (F = 7.55, df= 2, p< 0.0119), RCR (F = 2.18, df= 2, p < 0.169) and RGR (F = 6.96, df = 2, p < 0.0149) have been reduced while AD (F = 1.61, df = 2, p < 0.2534) in treated larvae showed increase compared with the control.

The effectiveness of plant essential oil on digestive enzymes under LC50 (1.528%) concentration after 24 and 48 h of treat-ment was analyzed (Tables 4 and 5). The amount of protein was decreased significantly compared with the control. It was lowest in the larvae treated with Origanum 24 h after treatment (38.72 ± 5.10 OD/min). The amount of triglycerides decreased significantly compared with the control 48 h after treatment (0.918 ± 0.03 mg/mL). Activity of a-amylase enzyme was decreased significantly compared with the control 48 h after

treatment (0.422 ± 0.039 lmol/min/mg protein). The activity of alkaline phosphatase and protease in treated larvae decreased compared with the controls. Lipase was increased 24 h after treatment (0.297 ± 0.011 nmol/min/mg protein) compared with the control (0.239 ± 0.002 nmol/min/mg pro-tein). Significant differences in detoxifying enzymes such as glutathione S-transferase and esterase were noted compared to controls.

4. Discussion

Secondary organic compounds synthesized by plants have an important role in protecting plants against insect attacks. These compounds affect insects by way of being toxic, causing a delay in larval growth or acting as antifeedants (Isman, 2006). The present results clearly indicated insecticidal activity of Origanum essential oil on third instar larvae of diamond back moth in a dose dependent rhythm. Essential oils of many plants have been reported to act as toxicants in several insect species and by several means; as fumigants, topical and contact (see review byRegnault-Roger et al. (2004)).

Deterrence is another characteristic present in essential oils (EOs) used for controlling insects. The main reason for deter-rence comes from the effects on olfactory senses of insects set by essential oils. The individual chemical/s preset in EOs may act as deterrent or their combined effects may have increasing effect on deterrence (Gillij et al., 2008; Hummelbrunner and Isman, 2001). In the present study we observed higher deter-rence by the used EO in a dose dependent manner.

Sublethal concentrations (LC10 and LC30) of the essential

oil on feeding indices such as ECI, ECD, RCR and RGR showed reduction compared with the controls. However, a slight increase in AD in treated larvae was observed, although this increase was not significant compared with the control. Nutritional indices are used for determination of growth and/or feeding inhibition. This finding indicates that during treatment, food remains longer in insect gut. Similar results are reported by Mordue and Blackwell (1993), Senthil Nathan and Sehoon (2006). ECI is considered as a tool to esti-mate the efficiency of food being ingested to be converted for

Table 1 Estimated concentration LC50, LC30, LC10of Origanum essential oil.

Essential oil %LC10 %LC30 %LC50

Confidence limit 95% Confidence limit 95% Confidence limit 95%

Origanum vulgare 0.234 (0.1089–0.3716) 0.710 (0.2076–0.5466) 1.528 (1.1543–2.0732)

Table 2 The mean percentage repellency ± standard error of essential oil.

Concentration Origanum vulgare P-Value

%LC10 22.8 ± 6.64b 0.048

%LC30 49.8 ± 6.95a

Table 3 The effect of sublethal concentrations (LC10and LC30) O. vulgare on feeding efficiency.

Essential oil AD% ECD% ECI% CI RGR

mg/mg/insect mg/mg/insect

Control 51.74 ± 4.34a 17.36 ± 2.95a 8.51 ± 0.93a 32.73 ± 1.39a 2.83 ± 0.40a

%LC10 55.83 ± 4.06a 11.89 ± 1.06ab 6.55 ± 0.16ab 32.66 ± 1.78a 2.14 ± 0.15ab

%LC30 61.66 ± 3.52a 9.22 ± 0.26b 5.68 ± 0.43b 29.52 ± 0.51a 1.67 ± 0.13b

Within columns, means followed by the same letter do not differ significantly (p > 0.05).

AD – approximate digestibility; ECD – efficiency of conversion of digested food; ECI – efficiency of conversion of ingested food; CI – consumption index; RGR – relative growth rate.


growth. A reduction in this index shows that most part of the ingested food is being metabolized for energy rather than being converted to body mass. Reduction in ECD indicates the inability of the insect to convert digested food for growth. In fact the energy of digested material might have been diverted into detoxification process (Silveira Ramos et al., 2009). The RGR and RCR reduction may be an indication of damages caused by allelochemicals present in the essential oil to per-itrophic membrane or cell surfaces in the mid gut (Marie et al., 2009). In physiological studies, determination of total protein and many of chemical macromolecules such as lipids and carbohydrates are very important. Many of insecticides have antifeedant properties and reduce feeding efficiency in insects, which in turn reduces some of the vital components such as proteins in the body (Etebari et al., 2006). In the pre-sent investigation, the amount of total protein decreased after treatment with LC50 dose of the essential oil however, this

amount increased after 48 h indicating more consumption of food material after an initial lapse of time. The initial decrease in protein could be due to the breakdown of proteins into amino acids and their entry into the TCA cycle as keto acid for the compensation for lower energy caused by chemical stress (Schoonhoven, 1982).

Alpha-amylase is an enzyme hydrolyzing starch to maltose and glycogen to glucose. This enzyme was reduced in treated larvae. Senthil Nathan and Kalaivani (2005) reported its reduction in Cnaphalocrocis medinalis (Gnenee) under the effect of neem extract, leaf extracts of Vitex negundo and the B. thuringiensis.Shekari et al. (2008) also reported its reduc-tion in the larvae of Elm leaf beetle Xanthogaleruca luteola

Mu¨ller (Col: Chrysomelidae) with sweet wormwood

Artemisia annuaL. (Asterales: Asteraceae) methanolic extract after 24 h and its increase after 48 h. Similarly Zibaee and Bandani (2009) reported its reduction in shield bug Eurygaster integriceps Puton under the effect of A. annua extract.

Hydrolysis of peptide bonds in insects is depended on a group of enzymes called proteases. In this study protease

activity was reduced in treated larva although not significant. This result is consistent with other reports where reduction in protease is mostly reported (Liu et al., 2008; Zhang and Chiu, 1992).

Lipases play a major role in storage and lipid mobilization. These enzymes are also the basic components in many physio-logical processes such as reproduction, growth, and defense against pathogens. In our study, lipase activity was enhanced 24 h after treatment compared with the control but it was decreased 48 h after treatment.Senthil Nathan (2006)showed that larvae of Cnaphalocrosis medinalis Guenee treated with neem had low lipase activity in the midgut. Zibaee and Bandani (2009) also reported low lipase activity in Eurygaster integriceps treated by A. annua extract. The increase in lipase could be attributed to its esterase activity needed for detoxification. However, the later reduction is due to the inhibitory activity inserted by the chemicals present in the essential oil of Origanum.

ALP is hydrolytic enzymes that hydrolyzes phosphomo-noesters at alkaline pH. Toxic chemicals, on the other hand, decrease the nutrition efficiency and ALP activity (Yoshitake et al., 1966; Eguchi and Iwamoto, 1975). Senthil Nathan (2006) showed that treatment of rice plants with Melia azaderachJuss (Meliaceae) extracts decreased the activity level of ALP in Cnaphalcrocis medinalis. These authors reported that feeding Spodoptera litura Fabricius, on Ricinus communis L. treated with azadirachtin decreased the amount of this enzyme in the midgut (Senthil Nathan and Kalaivani, 2005). Our results show that the activity level of this enzyme decreases at 24 h after treatments but sharply increases at 48 h. Increasing ALP activity level may indicate perhaps that the enzyme is involved in detoxification processes.

Glutathione S-transferases (GST) are mainly cytosolic enzymes that catalyze the conjugation of electrophile mole-cules with reduced glutathione (GSH), potentially toxic sub-stances become more water soluble and generally less toxic (Grant and Matsumura, 1989). GSTs play an important role in insecticide resistance and are involved in the metabolism

Table 4 Effects of O. vulgare on some biochemical compounds of P. xylostella 3rd instar larvae (24 h after treatment).

Essential oil Protein (od/min) Triglyceride (mg/ml) a-Amylase (lmol/min/ mg protein) Lipase (nmol/min/mg protein) Protase (od/min) Alkaline phosphatase (lmol/min/mg protein) GST (lmol/min/mg protein) Esterase (lmol/min/mg protein) Control 49.12 ± 1.63a 1.29 ± 0.15a 2.15 ± 0.03a 0.239 ± 0.002b 35.32 ± 3.01a 2.24 ± 0.36a 4.44 ± 0.021a 57.08 ± 0.76a O. vulgare 38.72 ± 5.10b 1.01 ± 0.06a 1.6 ± 0.21a 0.297 ± 0.011a 14.03 ± 0.21b 1.62 ± 0.35a 3.21 ± 0.53a 6.62 ± 0.051b Within columns, means followed by the same letter do not differ significantly (p > 0.05).

Table 5 Effects of O. vulgare on some biochemical compounds of P. xylostella 3rd instar larvae (48 h after treatment).

Essential oil Protein (od/min) Triglyceride (mg/ml) a-Amylase (lmol/min/mg protein) Lipase (nmol/min/mg protein) Protase (od/min) Alkaline phosphatase (lmol/min/ mg protein) GST (lmol/min/mg protein) Esterase (lmol/min/ mg protein) Control 62.59 ± 2.81a 1.28 ± 0.028a 2.91 ± 0.15a 0.148 ± 0.012a 27.32 ± 2.86a 3.97 ± 0.21a 15.53 ± 0.106a 41.87 ± 0.51a O. vulgare 47.48 ± 1.07b 0.918 ± 0.03b 0.422 ± 0.039b 0.126 ± 0.011a 17.35 ± 1.35a 2.25 ± 0.007b 8.15 ± 0.42b 9.79 ± 0.76b Within columns, means followed by the same letter do not differ significantly (p > 0.05).


of organophosphorus and organochlorine compounds. Other xenobiotics such as plant defense allelochemicals against phy-tophagous insects induce GST activity (Yu, 1982; Vanhaelen et al., 2001). In this study, activity level of GST at 24 and 48 h after treatment decreased significantly compared with control. This result indicates its inhibition by essential oil more significantly after 24 h but the increase after 48 h which is still remains lower than the control may show that the essential oil loses its activity to some extend after 48 h of treatment. Esterase (EST) is an important detoxifying enzyme which hydrolyzes the esteric bond in synthetic chemicals. Also, ester-ase is one of the enzymes showing the strongest reaction to environmental stimulation (Hemingway and Karunatne, 1998). In this study EST significantly decreased compared to control 48 h after treatment. Similar results were reported by Liu et al. (1990).

5. Conclusions

In conclusion, the present results indicates that O. vulgare essential oil possess larvicidal effects on P. xylutella and in sub-lethal concentrations acts as antifeedant. The irreversible effects on metabolism and enzymes may nominate it as a

potent candidate for search in environment friendly

insecticide. Conflict of interest

The authors have no conflict of interest, involvement, financial or otherwise, that might potentially bias our work.


Financial assistance from the research department, University of Guilan, during this research program is gratefully acknowledged.


Ankersmit, G.W., 1953. DDT resistance in Plutella maculipennis(Curt.) (Lepidoptera) in Java. B. Entomol. Res. 44, 421–425.

Anonymous, 1996. Diamondback moth. Canada’s Green Plan. <http://www.agr.gov.sk.ca/docs/cropsl>.

Bernfeld, P., 1955. Amylases, a and b. Method Enzymol. 1, 149–158.

Bessey, O.A., Lowry, O.H., Brock, M.J., 1946. A method for the rapid determination of alkaline phosphatase with five cubic millimeters of serum. J. Biol. Chem. 164, 321–329.

Bradford, M., 1976. A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254.

Eguchi, M., Iwamoto, A., 1975. Changes in protease, esterase and phosphatase in the alimentary canal of the silkworm during metamorphosis. Insect Biochem. 5, 495–507.

Etebari, K., Bizhannia, A.R., Sorati, R., Matindoost, L., 2006. Biochemical changes in haemolymph of silkworm larvae due to pyriproxyphen residue. Pestic. Biochem. Physiol. 88, 14–19.

Fossati, P., Prencipe, L., 1982. Serum triglycerides determined colorimetrically with an enzyme that produces hydrogen peroxide. Clin. Chem. 28, 2077–2080.

Garcia-Carreno, F.L., Haard, L., 1993. Characterization of protease classes in langostilla Pleuroncodes planipes and crayfish Pacifastacus astacusextracts. J. Food Biochem. 17, 97–113.

Gillij, Y., Gleiser, R., Zygadlo, J., 2008. Mosquito repellent activity of essential oils of aromatic plants growing in Argentina. Bioresource Technol. 99, 2507–2515.

Grant, D.F., Matsumura, F., 1989. Glutathione S-transferase 1 and 2 in susceptible and insecticide resistant Aedes aegypti. Pestic. Biochem. Physiol. 33, 132–138.

Habing, W.H., Pabst, M.J., Jakoby, W.B., 1974. Glutathione S-transferases. The first step in mercapturic acid formation. J. Biol. Chem. 249, 7130–7139.

Hemingway, J., Karunatne, S.H.P.P., 1998. Mosquito carboxylester-ases: a review of the molecular biology and biochemistry of a major insecticide resistance mechanism. Med. Veter. Entomol. 12, 1–12.

Hummelbrunner, A.L., Isman, M.B., 2001. Acute, sublethal, anti-feedant and synergistic effects of monoterpenoid essential oil compounds on the tobacco cut worm (Lepidoptera: Noctuidae). J. Agr. Food Chem. 49, 715–720.

Isman, M.B., 2000. Plant essential oils for pest and disease manage-ment. Crop Prot. 19, 603–608.

Isman, M.B., 2001. Pesticides based on plant essential oils for management of plant pests and diseases. In: International Symposium on Development of Natural Pesti-Cides from Forest Resources. Korea Forest Research Insti-tute, Seoul, Republic of Korea, pp. 1–9.

Isman, M.B., 2006. Botanical insecticides, deterrents, and repellents in modern agriculture and an increasingly regulated world. Ann. Rev. Entomol. 51, 45–66.

LeOra Software, 1987. Polo-Pc a User’s Guide to Probit or Logit Analysis. LeOra Software, Berkeley, California, p. 22.

Lingathurai, S., Ezhil Vendan, S., Gabriel Paulraj, M., Ignacimuthu, S., 2011. Antifeedant and larvicidal activities of Acalypha fruticosa Forssk. (Euphorbiaceae) against Plutella xylostella L. (Lepidoptera: Yponomeutidae) larvae. J. K. S. U. (Science) 23, 11–16.

Liu, Y., Alford, A.R., Rajab, M.S., Bentley, M.D., 1990. Effects and modes of action of citrus limonoids against frcma ci Aiit fcemlin-eata. Physiol. Entomol. 15, 45–37.

Liu, C.H., Mishra, A.K., Tan, R.X., Tang, C., Yang, H., Shen, Y.F., 2006. Repellent and insecticidal activities of essential oils from Artemisia princepsandand Cinnamomum camphoraand their effect on seed germination of wheat and broad bean. Bioresource Technol. 97, 1969–1973.

Liu, Z.L., Ho, S. Hung, Goh, S. Hock, 2008. Effect of fraxinellone on growth and digestive physiology of Asian corn boree, Ostrinia furnacalisGuenee. Pestic. Biochem. Physiol. 91, 122–127.

Marie, S.S., Amr, E.M., Salem, N.Y., 2009. Effect of some plant oils on biological, physiological and biochemical aspects of Spodoptera littoralis(Boisd.). J. Agri. Biol. Sci. 5, 103–107.

Mordue, A.L., Blackwell, A., 1993. Azadirachtin: an update. J. Insect Physiol. 39, 903–924.

Neto Bandeira, G., Augusto Gomes da Camara, C., Martins de Moraes, M., Barros, R., Muhammad, S., Akhtar, Y., 2013. Insecticidal activity of Muntingia calabura extracts against larvae and pupae of diamondback, Plutella xylostella (Lepidoptera, Plutellidae). J K. S. U. (Science) 25, 83–89.

Regnault-Roger, C., Ribodeau, M., Hamraoui, A., Bareau, I., Blanchard, P., Gil-Munoz, M.I., Barberan, F.T., 2004. Polyphenolic compounds of Mediterranean Lamiaceae and inves-tigation of orientional effects on Acanthoscelides obtectus(Say). J. Stored Prod. Res. 40, 395–408.

Sarfraz, M., Keddie, B.A., 2005. Conserving the efficacy of insecticides against Plutella xylostella (L.) (Lep., Plutellidae). J. Appl. Entomol. 129, 149–157.

Sarfraz, M., Dosdall, L.M., Keddie, B.A., 2005. Spinosad: a promising tool for integrated pest management. Outlooks Pest Man. 16, 78– 84.

SAS Institute, 1997. SAS/STAT User’s Guide for Personal Computers. SAS Institute, Cary, Nc.


Schoonhoven, L.M., 1982. Biological aspects of antifeedants. Entomol. Exp. Appl. 31, 57–89.

Senthil Nathan, S., 2006. Effects of Melia azedarachon nutritional physiology and enzyme activities of the rice leaffolder Cnaphalocrocis medinalis (Gnenee) (Lepidoptera: Pyralidae). Pestic. Biochem. Physiol. 84, 98–108.

Senthil Nathan, S., Kalaivani, K., 2005. Efficacy of nucleopolyhe-drovirus (NPV) and azadirachtin on Spodoptera litura Fabricius (Lepidoptera: Noctuidae). Biol. Control 34, 93–98.

Senthil Nathan, S., Sehoon, K., 2006. Effect of Melia azaderach L. extract on the teak defoliator Hyblaea puera Cramar (Lep: Hyblaeidae). Crop Prot. 25, 287–291.

Shekari, M., Jalali Sendi, J., Etebari, K., Zibaee, A., Shadparvar, A., 2008. Effects of Artemisia annua L. (Asteracea) on nutri-tional physiology and enzyme activities of elm leaf beetle, Xanthogaleruca luteola Mull. (Coleoptera: Chrysomellidae). Pestic. Biochem. Physiol. 91, 66–74.

Shelton, A.M., 2004. Management of the diamondback moth: de´ja ‘vu all over again? In: Endersby, N.M., Ridland, P.M. (Eds.), The Management of Diamondback Moth and Other Crucifer Pests. Proceedings of the Fourth International Workshop, 26/29 November 2001, Melbourne. Department of Natural Resources and Environment, Melbourne, Australia, p. 3/8.

Shelton, A.M., Sears, M.K., Wyman, J.A., Quick, T.C., 1983. Comparison of action thresholds for lepidopterous larvae on fresh market cabbage. J. Econ. Entomol. 76, 196–199.

Silveira Ramos, V., Gracas Machado Freire, M., Roberto Postali Parra, J., Ligia Rodrigues Macedo, M., 2009. Regulatory effects of an inhibitor from Plathymenia foliolosaseeds on the larval devel-opment of Anagasta kuehniella (Lepidoptera). Comp. Biochem. Physiol. 152, 255–261.

Singh, G., Upadhyay, R.K., 1993. Essential oils: a potent source of natural pesticides. J. Sci. Ind. Res. 52, 676–683.

Smith, C.M., Khan, Z.R., Pathak, M.D., 1994. Techniques for Evaluating Insect Resistance in Crop Plants. CRC Press, Florida, USA, p. 319.

Tabashnik, B.E., 1994. Evolution of resistance to Bacillus thuringiensis. Annu. Rev. Entomol. 39, 47–79.

Talekar, N.S., Shelton, A.M., 1993. Biology, ecology, and management of the diamondback moth. Annu. Rev. Entomol. 38, 275–301.

Tsujita, T., Ninomiya, H., Okuda, H., 1989. P-Nitrophenyl butyrate hydrolyzing activity of hormone-sensitive lipase from bovine adipose tissue. J. Lipid Res. 30, 997–1004.

Van Asperen, K., 1962. A study of housefly esterases by mean of sensitive colorimetric method. J. Insect Physiol. 8, 401–416.

Vanhaelen, N., Haubruge, E., Lognay, G., Francis, F., 2001. Hoverfly glutathione S-transferases and effect of Brassicaceae secondary metabolites. Pestic. Bioch. Physiol. 171, 170–177.

Verkerk, R.H.J., Wright, D.J., 1996. Multitrophic interactions and management of the diamondback moth: a review. B. Entomol. Res. 86, 205–216.

Waldbauer, G.P., 1968. The consumption and utilization of foods by insects. Adv. Insect Physiol. 5, 229–288.

Wink, M., 1993. Production and application of phytochemicals from an agricultural perspective. In: van Beek, T.A., Breteler, H. (Eds.), Phytochem. Agri., vol. 34, pp. 171–213.

Yazdani, E., Jalali Sendi, J., Aliakbar, A., 2013. Chemical composi-tion, toxicityand physiological effects of essential oil of Rosemarinus officinalison lesser mulberry pyralid, Glyphodes pyloalisWalker (Lepidoptera: Pyralidae). Crop Prot. 2, 461–476.

Yoshitake, N., Eguchi, M., Akiyama, A., 1966. Genetic control on the alkaline phosphatase of the midgut in the silkworm. J. Ser. Sci. Jpn. 35, 1–6.

Yu, S.J., 1982. Host plant induction of glutathione S-transferase in the fall armyworm. Pestic. Biochem. Physiol. 17, 52–60.

Zhang, X., Chiu, S.F., 1992. Effects of toosendanin on several enzyme systems of the cabbageworm Pieris rapae L. Acta Entomol. Sin. 35, 171–177.

Zibaee, A., Bandani, A.R., 2009. Effects of Artemisia annua L. (Asteracea) on the digestive enzymatic profiles and the cellular immune reactions of the sun pest, Eurygaster integriceps (Heteroptera: Scutellaridae), against Beauvaria bassiana. B. Entomol. Res. 12, 1–11.


Related documents

The hybrid phage, helper phage and hyper phage 21 enable engineered phage to display large folded therapeutic proteins along with phage coat proteins as fusion pro- teins with

Centers for Disease Control and Prevention; EHR: Electronic health record; EMR: Electronic medical record; GHC: Group health centre; ITS: [Niagara Region] information

This study demonstrates that salivary uric acid is also elevated in patients with metabolic syndrome and correlates with several cardiometabolic risk factors including blood pres-

(5) The fly ash residues (including unreacted or undis- solved particles, particles under alkaline attack and reaction products) were collected after different soaking times and

Il est possible que d’autres pompes a efflux, comme les transporteurs ABCG5 et G8 (Iqbal et Hussain, 2009), initialement impliqu es dans le transport inverses du cholest erol et

Illustration 8 shows a small hair comb from Maison Auguste Bonaz which exemplifies this new style.. In its simple asymmetric shape it reflects the influence of popular art

Secondly, as a woman who has used self -tracking apps in the past myself, I felt in a better position to orient myself within the research and to speak on a more personal and