BIHAREAN BIOLOGIST 8 (2): 106-108 ©Biharean Biologist, Oradea, Romania, 2014 Article No.: 141119 http://biozoojournals.ro/bihbiol/index.html

Contact toxicity of the essential oils from Salvia leriifolia Benth (Lamiaceae) against serricorne (F.)

Jafar GERANMAYEH1 and Seyed Mehdi HASHEMI2,*

1. Department of Plant Protection, Faculty of Agriculture, Urmia University, Urmia, West Azerbaijan, Iran. 2. Young Researchers and Elite Club, Ardabil Branch, Islamic Azad University, Ardabil, Iran. * Corresponding author, S.M. Hashemi, E-mail: [email protected]

Received: 10. August 2013 / Accepted: 28. April 2014 / Available online: 18. July 2014 / Printed: December 2014

Abstract. Salvia leriifolia Benth (Lamiaceae) was collected at tow developmental stages. Dry ground leaves were subjected to hydrodistillation using a Clevenger-type apparatus and the chemical composition of the volatile oils was studied by gas chromatography-mass spectrometry (GC-MS). Contact toxicity of oils was investigated against adults of cigarette Lasioderma serricorne (F.). Both oils in the same concentrations were tested for their toxicity on pest. The mortality of 3-10 days old adults increased with rates of oils and exposure time. Results showed that May oil were more toxic than March oil against cigarette beetle. Contact effects of these essential oils were considered to warrant further research into their potential for commercial use.

Keywords: Salvia leriifolia, essential oil, contact toxicity, Lasioderma serricorne.

Introduction Materials and methods

During few recent decades and mostly because of their di- Plant material Fresh leaves of S. leriifolia were collected at two different times of the versity, versatility and safety in comparison with the syn- year. The first collection was done during March 2011, when growth thetic materials, natural products of plant, and mi- rates were maximal. Later collection was done during May 2011, in croorganism origins have attained special interest among full bloom. Plant materials were air dried for 7 days at room tem- academic and industrial scientific communities. Plant king- perature. Plants were harvested from the same location of Estaj (35° dom with an estimated population of about 500,000 species 55' latitude; 57° 37' longitude; 1700 altitude), a village located in the of which only half is described so far is among the most at- Razavi Khorasan Province, Northeast of Iran, 250 km of Mashhad. tractive pools of bioactive natural products (Colegate & Mo- Plant taxonomists in the Department of Biology at Urmia University, confirmed the taxonomic identification of plant species. Voucher lyneux 2008). specimens were deposited at the Herbarium (Voucher No. 1511) of Biological activities are correlated to the presence of the Department of Horticulture, Urmia University, Urmia, Iran. chemical compounds, particularly secondary metabolites. The presence of these compounds may assist in predicting Extraction and analysis of essential oil some traditional uses of medicinal plants (Rasoanaivo & Essential oil was extracted from the plant samples using a Ratsimamanga-Urverg 1993). Clevenger-type apparatus where the plant material is subjected to Salvia leriifolia (Benth) is a perennial herbaceous plant hydro-distillation. Conditions of extraction were: 100 g of air-dried sample; 1:10 plant material/water volume ratio, 2.5 h distillation. that grows exclusively in south and tropical regions of Anhydrous sodium sulphate was used to remove water after extrac- Khorasan and Semnan provinces, Iran. This plant was intro- tion. Extracted oil was stored in a refrigerator at 4 °C. duced in Florica Iranica in 1982 and has different vernacular The constituents of the essential oil were analyzed by gas chro- names such as Nuruozak and Jobleh (Rechinger 1982). In the matography–mass spectrometry (GC–MS) using a Shimadzu-9A gas aspect of medicinal properties, aerial parts of S. leriifolia have chromatograph equipped with a flame ionization detector (FID) and been used as relive pain, decrease blood sugar and treat in- with PH-5 capillary column (30 m × 0.1 mm; 0.25 μm film thickness). flammation and insomnia (Andalib et al. 2011). Researchers The oven temperature was held at 60 °C for 3 min, programmed at 20 °C /min to 240 °C and then held at this temperature for 8.5 min. have reported analysis of essential oil from S. leriifolia The carrier gas was Helium (99.999%) in a flow rate of 1 ml/min. (Rustaiyan 2000, 2007, Loizze et al. 2010, Monfared & Mass spectra was taken at 70 ev. The injector temperature was 280 Ghorbanli 2010, Mohammadhosseini 2012, Hosseini et al. °C. Identification of the constituents of the oils was made by com- 2013). Essential oils of S. leriifolia have been shown to possess parison of their mass spectra and Retention Indices (RI) with those biological activities, including in vitro treatment of Alz- given in the literature and those authentic samples (Adams, 1995). heimer's disease (Loizze et al. 2009), antiproliferative activity against human tumor cell in vitro models (Loizze et al. 2010) Insect culture Lasioderma serricorne was reared on wheat flour. Adult , 3–10 and antibacterial effect of essential oil obtained in different days old, were used for toxicity tests. They were reared in glass jars phenologic stages against cariogenic bacteria Hoshyar Sar- 2.5 L in volume, which were covered by a fine mesh cloth for ventila- jami et al. 2011). The insecticidal activity of essential oil from tion. The cultures were maintained in dark in a growth chamber set S. leriifolia has been evaluated against granary weevil, Sito- at 27 ± 2 C and 60 ± 5% RH. Parent adults were obtained from labo- philus granarius (L.), and lesser grain borer Rhyzopertha do- ratory stock cultures maintained at the Entomology Department, minica (F.) (Hosseini et al. 2013). University of Urmia, Iran. All experiments were carried out under According to our best knowledge, there are not reports the same environmental conditions. In bioassays, insects were con- sidered dead when no leg or antennal movements were observed on the contact insecticidal activity of S. leriifolia essential oil when gently prodded with a brush. against cigarette beetle Lasioderma serricorne (F.) so far. Thus, the present research reports the contact activity of essential Contact activity oil of S. leriifolia at two different times of the year against L. The contact activities were determined in impregnated-paper assays. serricorne. Appropriate amounts of both oils in same concentrations were dis- Salvia leriifolia on Lasioderma serricorne 107 solved in 200 μl acetone and applied to (Whatman No. 2, 4.25 cm di- Table 1. Chemical composition of the essential oil ameter) which were dried in air for 5 min. Controls received 200 μl of S. leriifolia in March and in May. acetone. Each filter paper was placed in the bottom of a vial (5 cm Composition Composition diameter and 3.5 cm height), and then 10 adults of insects (3-10 days Compound (%) in March (%) in May old) were placed in each vial and covered with a lid. Five replicates α-Pinene 15.89 12.01 were prepared for each treatment, and were kept under the same Camphene 2.56 3.65 environmental conditions described for the rearing. Mortality was β-Pinene 16.94 21.09 recorded after 3, 6, 9, 12 and 24 h of exposure to the essential oils. Δ-3-Carene 1.18 1.90

ρ-Cymene 0.97 1.06 Data analysis The mortality data were corrected using Abbott’s formula (Abbott 1,8-Cineole 20.36 34.76 1925) for the mortalities in the controls. Percentage mortality values Borneol 7.97 6.48 for different exposure times were subjected to analysis of variance Terpinen-4-ol 1.72 2.24 according to the general linear model (GLM) using the SPSS (1994) α-Terpineol 3.29 2.64 statistical program analysis. Data were transformed using arcsine √x α-Muurolene 1.69 1.51 to meet normality, before ANOVA. Means (±SE) of untransformed y -Cadinene 6.89 1.59 data are reported. Comparison of means was done through Tukey Δ -Cadinene 2.54 1.03 (HSD) test (α = 0.05). 10-epi-gamma-eudesmol 1.40 0.35 α-Cadinol 4.42 0.59

Results was investigated (Hosseini et al. 2013). In that experiment,

essential oil of leaves was evaluated at two different phono- Yield and essential oils composition logic stages (vegetative and flowering). In flowering time, The extracted oils was yellow in color, had an aromatic odor. the plant showed more fumigant activity than other The essential oils yield varied with the respective seasons phenologic stage. We observed that toxicities of the same (Table 1). The yield in March (0.48%, w/w) is higher than in plant collected at two different periods of the year showed May (0.29%, w/w). The compositions of sage essential oil significant variation, which could be attributed to changes in collected at different times of the year are shown in Table 1. the amounts of the active compounds present in each plant The different components of the oils are listed according to at different seasons. Plant material collected at different the increasing retention times. The components that varied times of the year may contain other possible novel com- seasonally included 1, 8-cineole and β-pinene (highest in pounds with other bioactivities (Eloff 1999). May: 34.76 and 21.09%, respectively), y-cadinene and α- The toxic effect of essential oils, apart from the variabil- cadinol (highest in March: 6.89 and 4.42%, respectively). ity of phytochemical patterns, involves several other factors. These components represent up to 58.04% of the total oil in The point of entry of the toxin is one of them. Commonly, May and 48.63% of the total oil in March (Table 1). essential oils can be skin absorbed (contact effect), inhaled Contact activity (fumigant effect), or digestive system (ingestion effect) by in- The toxicity of plant essential oils used against insects ex- sects (Regnault-Roger 1997). Insects have been shown to be posed to direct contact are recorded in Tables 2 and 3. Re- very sensitive for contact application of essential oils; here sponses varied according to plant stages, different rates, and experimental insect are not an exception (Su 1991, Huang et exposure times. May oil was relatively more toxic to the in- al. 1997). It is not surprising that strong activity was ob- sect species. At the lowest rate (4.66 mg/cm2) of the March served in the oils extracted from S. leriifolia, when applied oil yielded 100% mortality after a 12 h exposure. By contrast, topically to insects, if we assume 1, 8- cineole is the major the lowest rate (4.66 mg/cm2), of the May oil yielded 100% compound involved. 1, 8-cineole is also the main compound mortality after a 9 h exposure. At the highest rate (4.80 (58.49%) present in the essential oil of Callistemon viminalis mg/cm2) with the March oil, kills reached 86% with a 3 h (Myrtaceae), which showed high insecticidal activity when exposure. By contrast, at this rate and exposure with May applied topically on Acanthoscelides obtectus and Calloso- oil, 96% mortality were achieved. bruchus maculatus adults (Ndomo 2010), S. granarius, Sitophi- lus zeamais, Tribolium castaneum, and Prostephanus truncatus adults (Obeng-Ofori et al. 1997), and C. maculatus, Rhyzoper- Discussion tha dominica (F.), and Sitophilus oryzae adults (Acgarwal et al. 2001). The yields and chemical compositions of the essential oils 1, 8-cineole characterized as the main component (20.36 from S. leriifolia varied in March and in May. Researchers and 34.76%) from S. leriifolia essential oil in March and in have reported seasonal variation in essential oil yields and May, respectively. In contact with insects, 1, 8-cineole act by chemical constituents from the genus salvia (Pitarevic et al. blocking the synthesis of juvenile hormones. It inhibits ace- 1984, Farhat et al. 2001, Kamatou et al. 2008, Hosseini et al. tylcholinesterase by occupying the hydrophobic site of en- 2013). zyme’s active centre (Obeng-Ofori et al. 1997). This Many studies have reported differences in the biological monoterpenoid might have broad insecticidal activity activities from the genus Salvia collected in different seasons against stored product insects (Shaaya et al. 1991, Acgarwal (Farhat et al. 2001, Kamatou et al. 2008, Hoshyar Sarjami et et al. 2001, Lee et al. 2004, Rozman et al. 2007). So the toxic al. 2011, Hosseini et al. 2013). Insecticidal activity of S. leriifo- effect of S. leriifolia oil could be attributed to this component. lia essential oil against S. granarius and R. dominica and then In conclusion, our study demonstrated that the essential relation between insecticidal property and plant fenology oil from S. leriifolia have contact toxicity against L. serricorne.

108 J Geranmayeh & S.M. Hashemi

Table 2. Contact toxicity of essential oil from Salvia leriifolia in March (vegetative stage) against Lasioderma serricorne.

Mean mortality (%) Treatments Dose Exposure time (h) Insect (mg/cm2) 3 6 9 12 24 L. serricorne 4.66 40±3.5b 62±3.7b 74±3.2b 100±0b 100±0b 4.67 60±3.1ab 84±2.0ab 98±0.4ab 100±0ab 100±0ab 4.69 66±3.0a 88±1.7a 100±0a 100±0a 100±0a 4.72 74±3.2a 90±1.7a 100±0a 100±0a 100±0a 4.80 86±1.6a 100±0a 100±0a 100±0a 100±0a

1. Each datum represents the mean of five replicates, each set up with 10 adults (n=50). 2. Means within a column followed by the same letter are not significantly different at P = 0.05.

Table 3. Contact toxicity of essential oil from Salvia leriifolia in May (flowering stage) against Lasioderma serricorne.

Mean mortality (%) Treatments Dose Exposure time (h) Insect (mg/cm2) 3 6 9 12 24 L. serricorne 4.66 68±3.1b 86±2.0b 100±0b 100±0b 100±0b 4.67 70±3.9a 92±0.8a 100±0a 100±0a 100±0a 4.69 82±2.4a 98±0.4a 100±0a 100±0a 100±0a 4.72 90±1.2a 100±0a 100±0a 100±0a 100±0a 4.80 96±0.9a 100±0a 100±0a 100±0a 100±0a

1. Each datum represents the mean of five replicates, each set up with 10 adults (n=50). 2. Means within a column followed by the same letter are not significantly different at P = 0.05.

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