197-206 - International Journal of Farming and Allied Sciences
Transcription
197-206 - International Journal of Farming and Allied Sciences
International Journal of Farming and Allied Sciences Available online at www.ijfas.com ©2015 IJFAS Journal-2015-4-3/197-206/ 31 March, 2015 ISSN 2322-4134 ©2015 IJFAS Study of chemical composition and antibacterial effects of essential oils of Stachys lavandulifolia Vahl., Salvia verticillata L., and Tanacetum polycephalum Schultz-Bip. on some microbial lineages Mohammad Mahdavi1, Mohammad Hassan Jouri*, Samanehsadat Mahzooni-Kachapi2 and Solyman Halimi’Jelodar3 1.Assistant Prof., Department of Natural Resources, Islamic Azad University, Nour Branch, Nour, Mazandaran, Iran 2.MS in Rangeland Management and membership of Young Researchers Club, Islamic Azad University, Noor Branch 3.Cultural deputy, Islamic Azad University, Nour Branch, Nour, Mazandaran, Iran Corresponding author: Mohammad Hassan Jouri ABSTRACT: Acceleration of antibiotics resistances, their side effects, and overusing of chemical preservers of foods, which some of them are suspicious to carcinogenic and teratogenic effects or remain toxic elements in body, and are fairly caused to be noted the extraction, herbal medicines, and natural antibacterial materials. This paper gets around to role of the essential oils of Stachys lavandulifolia Vahl., Salvia verticillata L., and Tanacetum polycephalum Schultz-Bip., as are greatly applied at traditional and modern medicines, on some bacteria such as Staphylococcus aureus and Escherichia coli. Hence, aerial organs of these species were phonologically collected at flowering period and after shrivelling in vitro setting, extracting of essence was done by Hydrodistilation. Compositions of the essential oils were identified and analysed using GC and GC/MS and by measuring the Retention Index and Mass spectrums. The antibacterial effects of essences were surveyed on two bacteria using tubed dilution method to determine the MIC and MBC. Acquired analysis showed that efficiency of the essences in Stachys lavandulifolia, Salvia verticillata, and Tanacetum polycephalum was 0.69, 0.92, and 1.09%, respectively. The essence of Stachys lavandulifolia and Salvia verticillata has the most antibacterial effects upon Escherichia coli and the least one on Staphylococcus aureus. The essential oil of Tanacetum polycephalum has more effects on Staphylococcus aureus than Escherichia coli. As a result, the essences of these herbs have considerable antibacterial effects and can be desirably replaced in lieu of synthetic antibiotics, which bacteria become more resistance apropos of them. Moreover, it is suggested to use the essences in the food industries as natural additive and healthy mat. Keywords: essential oils, Stachys lavandulifolia, Salvia verticillata, Tanacetum polycephalum, Escherichia coli, Staphylococcus aureus, antibacterial effects Intl J Farm & Alli Sci. Vol., 4 (3): 197-206, 2015 INTRODUCTION Nearly all cultures, from ancient times, have used plants as a source of medicine. In many developing countries, traditional medicine is still the mainstay of health care, and most of the drugs and cures used come from plants (Ndawonde, 2006). In fact from the start of life to the last breath, almost every aspect of human life is deeply associated with plants (Kumar, 2009). In recent years, great notation has been focused on herbal medicines and natural products (Albuquerque , 2007) as are therapeutically used for traditional communities in the world (Moshafi , 2009). It is because of the side effects of chemical drugs (Reynolds, 1996) and many people therefore are turning to herbal remedies, especially for minor ailments, and modem scientific medicine still depends on plants, and the knowledge gained from plants, for some essential drugs (Ndawonde, 2006; Ozkan , 2010). Therefore, the use of herbal remedies requires sufficient knowledge about the efficacy, safety and proper use of such products (Barbosa , 2010; Nori-Sharagh , 1999). Hence, it is necessary to have baseline data regarding the use of herbal remedies and to educate future health professionals about various aspects of herbal remedies (Reynolds, 1996). On one side, many diseases refer to food poisonings, especially bacterial toxicities (Mortazavi, 2009) and as food health, turning to remove the pathogenic factors from packed foods is important and needs to some modern approaches which have less side-effect on human body and health (Burt, 2004; Holley and Patel, 2005). Because overusing of chemical preservers in food industries are suspicious to carcinogenic and teratogenic effects or remain toxic elements in body (Akhondzadeh , 2007). On the other hand, accelerative demanding to use organic foods without any chemical preservers (Gandomi’Nasrabadi , 2008; Gurib-Fakim, 2006; Ghahraman, 1994) is caused to apply the natural products such as essences and herbal extractions as food protectors and antibacterial elements (Gandomi’Nasrabadi , 2008; ITC, 2012). Since these products are completely natural, they can create the least harm to the body and natural environments as well (Beuchat, 2001). Species of Stachys lavandulifolia is a species of Stachys genus from Labiatae family (Babakhanlo , 1998). The extraction of aerial part of the species is used in the traditional medicine of Iran such as treatment of infection, asthma, and, and agonal illness, especial rheumatism (Marriout , 2001). The herb beefs up the stomach (Babakhanlo , 1998) and is effective to reduce the discomposure (Rasooli, 2012; Rechinger , 1982), the digestive disorders (Mukherjee, 2009), and the genital tumours and cancer ulcers (Javidnia , 2006), as well. Salvia lerticillata species also belongs to Labiatae family and Salvia genus which is aromatic and perennial plant (Rezvanpanah , 2011). The herb is used to numerous subjects including empowering of body and anti-hysteria in pertussis disease, reinforcer of heart, facilitator of digestive operation, antipyretic and antiseptic matter, cutter of chronic cough, reductor of blood sugar, and consumption of its products can be activated circulator function (Ghahraman, 1988). The herb of Tanacetum polycephalum, as aromatic and perennial species, belongs to Tanacetum genus (Asteraceae) and it has antiseptic, analgesic, anesthetic, disinfective, expectorant, anti-cancer, anti-allergic, and anti-irritant properties, and can also reduce blood pressure (Barazandeh, 2003; Pauwels , 2011). In recent years, plentiful researches have been done on antimicrobial effects of diverse essences, extractions, and condiments that show highly abilities of these sorts of components to prevent the growth of pathogenic microorganisms and decomposing of food matters (Ababutain, 2011). Regarding this, the study on species of Stachys genus showed that sesquiterpene formed the most parts of all species essence (Chalchat , 2000; Flamini , 2005; Tepe , 2004). (Maleki , 2004) clarified the main and whole components of Stachys lavandulifolia and some its antibacterial effects. The antibacterial surveying of some species of Stachys genus showed that their essences have fair anti-inhibiting effects on gram- positive and negative bacteria (Dulger , 2005; Tavassoli , 2011). Camphor, as the main elements in the essence of different species of Salvia genus, has been reported by many researchers such as (Chalchat , 1998; Pharmacopoeia, 1983; Weyerstahl , 1999). 1-8 cineole as 50 % of essence composition in Salvia fruticosa and S. aramiensis was also reported by (Bayrak and Akgul, 1987) and (Demirci , 2002). The α-thujone and 1-8 cineole were the main elements of the essential oils in planted Salvia officinalis and the α-thujone, 1-8 cineole, camphor, and β-caryophyllene were the most composition of the essence in S. triloba (Longaray Delamare , 2007). Although (Strijack , 2004) proved the anti-inhibiting effects on Escherichia coli bacterium by the α-thujone and 1-8 cineole in the essence of S. fruticosa, other researchers studied different antibacterial effects of the Salvia genus, as well (Bouaziz , 2009; Vagionas , 2007; Vjera Bilusic Vundac , 2006). Many researches have also focused on different species of the Tanacetum genus so that monoterpene hydrocarbon as the basic elements of essence in Tanacetum fruticulosum has been reported by (Mos’hafi , 2009). (Askin Akpulat , 2005) have pointed out that the main formative essences in Tanacetum argyrophyllum were cisthujene, trans-thujene, and 1-8 cineole while in the Tanacetum parthenium were camphor, camphene, and p-cimene. Components of 1-8 cineole, α-Pinene, trans-pinocarveol, and Spathulenol in Tanacetum nitens and ingredients of 18 cineole, santolina triene, and cryzanthenone in Tanacetum argenteum were found as the main elements in their essences (Bagci and Kocak, 2010). (Skaltsa , 1999) have reported that the essence of Tanacetum parthenium has 198 Intl J Farm & Alli Sci. Vol., 4 (3): 197-206, 2015 robustly the antibacterial effects on gram bacterium as (Saha , 2013) have also reported same capability against gram- positive and negative bacteria from the Tanacetum polycephalum species. Furthermore, other researchers have been reported that different herbal essences had considerable inhibition effects on microorganisms which were contamination agents in food matters (Burt, 2004; Mehdizadeh and Razavi’Rouhani, 2009). Using of antimicrobial combinations from plants essences, as natural products which have anti-inhibiting effects, has increasingly paid attention regarding the day-by-day resistance of bacteria apropos of antibiotics deriving from microorganisms. Study around the medicinal plants help to enable future strategies for knowledge and new compounds, or even drugs, acquisition (Mos’hafi , 2009). Hence, the paper has concentrated upon quality and quantity of the essences in three species, such as Stachys lavandulifolia, Salvia verticillata, and Tanacetum polycephalum, in different climatic circumstances and their antibacterial effects as well. MATERIALS AND METHODS A. Collecting of herbs and extraction of essence In this study, the aerial parts of the species were incidentally collected at a time of flowering from natural areas located in the central Alborz (Iran) at altitude of 2400 m, and the freshly extracted herb was dried in the laboratory setting. A sample (100 g) of the aerial parts of the Stachys lavandulifolia, 80 g of the Salvia verticillata, and 65 g of Tanacetum polycephalum were extracted using a Clevenger apparatus through water distillation for 3 hours (Goncalves , 2010). In order for the essential oil not to be mixed with water, 1 mili-liter of pentane was poured into the storage inlet of the essential oil. Considering the moisture percentage, the essential output was measured in dry weight (W/W). The essential oil, when extracted, was collected and distilled using sodium sulphate, and kept at 4 °C until it was injected into GC (Rabbani , 2005; Weyerstahl , 1999). B. Essential analysis The extracted essential oil was first injected into the GC; then, the most suitable programing of thermal column was obtained for complete separation of the essential oil (Marilena , 2001; Monteiro , 2014). In addition, the relative percentage of each component was measured with respect to the peak level in the GC chromatogram (Chamorro , 2012). Then, the essential oil was analysed using GC/MS in order to identify its composition (Mukherjee , 2011). The components were identified using deterrence indices and mass spectrometry, and were compared with the standard compositions and the data in the mass database Wiley275.L (Adams, 2007). C. Examination of anti-bacterial effects of the herbs In the present study, the gram-positive bacterium of Staphylococcus aureus (PTCC1431) and the gram-negative bacterium of Escherichia coli (PTCC1399), which have been prepared from the Centre for Scientific and Industrial Research of Iran, were used to test and examine the anti-bacterial effects of the extracted essential oils (Mothana , 2013; Saharkhiz , 2008). Preparation of McFarland 0.5 standard The standard was prepared by mixing 99.5 ml of 1% BaCl2 (Barium Chloride) and 0.05 ml of 1% H2SO4 (Sulphuric Acid) (Sivropolulov , 1995). To compare the turbidity, which was 1×10 8 Colony Forming Unit (CFU) per ml, the standard was put into screw cap test tube (Tucker , 1996). Determination of MIC and MBC The Minimum Inhibitory Concentration (MIC) was defined as the lowest concentration of the compound to inhibit the growth of microorganisms (Sivropolulov , 1995); hence in order to determine the MIC, 1 milliliter Muller-Hinton broth culture was added to sterile test tube (9 tubes). Prepared suspension of bacteria with 10 8 CFU/ml consistency and 1 ml amount was added to each test tube (Tepe , 2005). The essential oils and extracts of the herbs were dissolved in DMSO (dimethyl-sulfoxide) regarding broth dilution method (proportional ½) and sightly densities were obtained by adding of 1 ml of each prepared dilution to test tubes covering bacterial culture substance and suspension. Ringer solution was used in the test tubes as positive (instead of the essence) and negative (in lieu of bacterial suspension) standard to determine the sensitivity of each bacterial species tested (Sivropolulov , 1995). The MIC values of the extracts against bacterial strains were determined based on the micro-well dilution method by means of incubating of the inoculated tubes at 37°C for 24 hours. In fact, turbidity of the substance inside of tubes demonstrates the bacterial growth and the MIC was determined when the first tube was entirely cleared without any turbidity. Minimum Bactericide Concentration (MBC) was determined in quite sterile condition of the test tubes without 199 Intl J Farm & Alli Sci. Vol., 4 (3): 197-206, 2015 any turbidity so that 500 µl of it was taken and put it to plates where nutrient agar culture substance had and then incubated at the appropriate temperature 37°C for 24 hours (Çetin , 2011; Fatouma , 2010). Ultimately, growth and non-growth of bacteria were followed and the first density, which did not have any bacterium growth, was considered as the MBC (Baron and Finegold, 1990). Primary concentrations of Stachys lavandulifolia (388 mg/ml), Salvia verticillata (350 mg/ml), and Tanacetum polycephalum (5×105 mg/ml) were prepared to 0.1 to 25 mg/ml of the essences concentration using dilution series method so that the highest and lowest concentrations were found in the first and ninth test tubes. Each assay in this study was repeated at least twice (Sivropolulov , 1995). RESULTS AND DISCUSSION A) Percentage and formed composition sort in the essences Essences of the herbs were yellow colour which was obtained in terms of dried weight (w/w) in this research so that the component of essential oils ranged according to the species (Table 1). Analysis and evaluation of the GC and GC/MS system showed that 58 components were identified, comprising 96.99% of the total components in the oil of Stachys lavandulifolia. The major components of S. lavandulifolia were Hexadecanoic acid (13.96%), alphapinene (13.74%), Germacrene D (8.95%), beta-pinene (7.01%), and beta-myrcene (4.48%). Oil of Salvia verticillata contained mainly trans-caryophyllene (18.82%), Germacrene D (9.49%), Spathulenol (7.53%), Sabinene (6.52%), and Bicyclo [3.1.1] heptane,6,6-dime (6%) from the 65 identified components composed of 96.05% of the total elements in the oil of the herb. On the other hand, 48 compounds were recognised representing 79.29% of the oil of the Tanacetum polycephalum. This oil is majored by 4,5-epoxy-1-isoprophyl-4-methyl-1 (18.42%), 1,8-cineole (14.55%), Borneol (16.66%), cryzanthenone (4.83%), and camphene (2.86%) as the important components in the essential oil of the plant. The essence efficiency averages to Stachys lavandulifolia, Salvia verticillata, and Tanacetum polycephalum were 0.69, 0.92, and 1.09%, respectively. Table 1. chemical compounds in the essences of Stachys lavandulifolia, Salvia verticillata, and Tanacetum polycephalum on central Alborz Mt Row Compound 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 Alpha-Amorphene Alpha-Cadinol Alpha.-Caryophyllene Alpha-Copaene Alpha.-Cubebene Alpha.-Gurjunene Alpha-Phellandrene Alpha-Pinene Alpha-Thujenal Alpha-Thujene Alpha-Terpinene Alpha-Terpineol Alpha- Terpinolene Anymol (3S,4R,5S,6R,7S)-Aristol-9-en-3… Aromadendrene Azulene,1,2,3,3a,4,5,6,7-octah Beta-Bourbonene Beta-Cubebene Beta-Elemene Beta-Myrcene Beta-Phellandrene Beta-Pinene Beta-Ocimene Y Beta-Bisabolene Beta-Costol Benzenemethanol, 4-(1-methylethyl) Bicyclogermacrene Bicyclo[4.4.0]dec-1-ene, 2-isoprop yl-5-methyl-9methylene Bicyclo[3.1.1] heptane, 6,6-dime.. Bicyclo[7.2.0]undec-4-ene-trimethyl-8-methylene Borneol Bornyl acetate 30 31 32 33 Stachys lavandulifolia 0.21 2.62 2.23 0.44 13.74 1.02 0.24 0.21 0.16 0.56 0.80 0.24 0.78 4.48 5.68 7.01 0.38 0.65 2.14 Salvia verticillata 0.21 1.78 5.81 0.35 0.21 0.12 1.77 2.20 0.23 0.80 0.40 1.24 0.13 2.66 Tanacetum polycephalum 1.65 0.10 0.10 0.38 0.74 0.38 0.08 - 0.37 0.88 - 0.11 6 0.70 - 0.29 0.12 16.66 - Retention Index 1514 1647 1691 1597 1375 1637 963 924 1184 907 1002 1190 1080 1690 1939 1916 1789 1383 1428 1390 932 1018 985 1039 1508 1862 1302 1500 1518 1142 1578 1168 1284 200 Intl J Farm & Alli Sci. Vol., 4 (3): 197-206, 2015 Row Compound 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 4-Bromo-1-naphthalenamine Cadina-1-4-diene Camphene Carvol Caryophyllene Caryophylla-4(12),8(13)-dien-5.beta.-ol Caryophyllenol-II Chrysanthenone Chrysanthenyl acetate 1,8-Cineole Cis-Ocimene Cis-Alpha-Bisabolene Cis- Davanone Cis-Jasmone Cis-Sabinene hydrate Compound 889 Cuminyl aldehyde 3-Cyclohexen-1-carboxaldehyde 3-Cyclohexen-1-ol, 4-methyl-1 Cyercene 4 Davana ether Delta-Cadinene Delta 3-Carene 4,4-Dimethl-3-(3-methylbut-3-e… 10,10Dimethyl-2,6-dimethylenebicy clo[7.2.0]undecan5.beta.-ol Docosane Eicosane 5-Epi-Neointermedeol (E)-1-(P-methoxyphenyl)-2-metho 4,5-Epoxy-1-isopropyl-4-methyl-1 Filifolone 2(3) - Furanone, 5-(2,5-dimethyl Gamma.-Cadinene Gamma.-Gurjunene Gamma.-Selinene Gamma-Terpinene Germacrene D 1H-Benzocyclohepten-7-oL , 2,3,4 2H-1-Benzopyran,3,4,4a,5,6,8 a 1H-Cycloprop[e]azulene, decahydro-1,1,7-trimethyl-4methylene Heptacosane 1,5-Heptadien-4-ol, 3,3,6-trimethyl Hexadecanoic acid 4-Hexadecen-6-yne,(z) Heptadecanoic acid 1(2H)-Naphthalenone, octahydro 7R ,8R-8-Hydroxy-4-isopropyliden 1-Hydroxy-1-methyl-7(methylethe… Iso-Bornyl acetate IsoCaryophyllene Isoledene Iso spathulenol Ledol Linalool L Linoleic acid Menthofuran 1-4 Methano-1H-indene, octahydro-1,7a-dimethyl-4(1-methylethenyl 4-Methyl-2-(3-methyl-2-butenyl)-furan Mintsulfide Myrtenol Naphthalene, 1,2,3,5,6,7,8,8a-octahydro-1,8adimethyl-7-(1-methylethenyl) Naphthalene,1 ,2,3,4-tetrahydro Nonadecane (CAS) 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 Stachys lavandulifolia 0.03 0.32 1.26 0.34 3.36 0.65 0.12 0.20 2.08 0.12 - Salvia verticillata 0.31 0.14 0.60 0.93 0.23 1.97 0.23 0.63 Tanacetum polycephalum 2.86 0.06 0.11 0.09 4.83 0.16 14.55 1.04 0.20 0.85 0.12 0.22 0.72 - - - 0.33 0.09 0.51 0.94 8.95 - 0.84 0.43 1.02 0.69 0.80 0.19 9.49 0.82 0.16 0.68 18.42 0.92 0.61 - 0.24 3.82 - 1.32 13.96 0.20 2.19 0.21 2.55 - 1.86 0.12 1.12 0.21 2.47 0.16 1.11 0.67 - 0.16 0.26 1.60 0.10 0.29 0.72 - - - 0.14 - 0.06 1.26 0.08 - - - 0.38 0.11 - Retention Index 1723 1771 1397 1249 1420 1640 1678 1129 1263 1021 1032 1542 1588 1402 1060 2545 1241 1857 1377 1603 1516 1523 990 1863 1635 1963 2293 1600 1818 1156 1098 1743 1494 1838 1897 1049 1487 1841 1502 1630 2695 1080 1995 1913 2074 1928 1971 1903 1286 1699 1705 1889 1851 1096 2149 1398 1609 1483 2001 1197 1621 1959 1490 201 Intl J Farm & Alli Sci. Vol., 4 (3): 197-206, 2015 Row Stachys lavandulifolia 1.29 0.32 0.41 1.37 1.16 0.46 3.44 0.22 0.72 0.55 1.14 0.54 0.33 0.51 96.99 Compound 97 Nonanal 98 Octadecane 99 1,3,6-Octatriene ,3,7-dimethyl 100 9,12,15-Octadecatrien-1-ol 101 1-Octen-3-OL 102 O-cymene 103 2-Oxibicyclo[4,4.0] dec-9-ene 104 PalatinolIC 105 Pentacosane 106 2-Pentadecanone, 6,10,14-trimethyl 107 1-Phellandrene 108 Phytol 109 Pinocarvone 110 Sabinene 111 Salvial-4(14)-en-1-one 112 Santolina triene 113 Spathulenol 114 Tau.-Muurolol 115 Terpinene-4-ol 116 Terpineol Z-beta 117 Tetracosane 118 Tetradecane ,4-methyl 119 Tetradecanoic acid 120 1,1,4,4-Tetramethyl-2-tetralone 121 2,3,4,5-Tetramethylthiophene 122 Trans-beta-Farnesene 123 Trans Carveol 124 Trans-Caryophyllene 125 Trans-Sabinene hydrate 126 Tricyclene 127 Tricyclo[3.3.2.0(3,7)] decane 128 1,5,5- Trimethyl-6-methylene-cyc 129 2,6,9,9-Trimethyltricyclo 130 Thymol 131 Valencene 132 Veridiflorol 133 Vulgarol 134 Yomogi alcohol Total compounds percentages Salvia verticillata 0.31 0.55 0.30 0.23 0.15 0.57 0.53 0.29 6.52 7.53 0.48 0.14 0.37 18.82 0.21 0.22 0.76 0.50 0.43 96.05 Tanacetum polycephalum 0.57 0.76 0.68 2.36 0.27 1.07 0.66 0.42 0.16 0.39 0.22 0.50 0.23 79.29 Retention Index 1305 2897 1222 2433 1162 1014 1501 1869 2493 1845 1178 2114 1164 1214 1595 873 1586 1894 1175 1106 3094 1541 1775 1679 1528 1455 1232 1655 1254 895 1880 1555 1797 1300 1656 1589 1943 1035 - B) The MIC and MBC outcome The effects of inhibitory and bactericidity of the essences on microorganisms were different (Table 2) so that the maximum demandable concentration to control the bacteria growth (MIC) and bactericidal rate (MBC) referred to the Staphylococcus aureus as for Stachys lavandulifolia species, and the minimum one was to Escherichia coli referring to Salvia verticillata species. On the other hand, the species of S. verticillata had the most MBC and MIC on S. aureus bacteria while S. lavandulifolia had reversely had the least effects on this bacterium. The herb of S. verticillata had also the most MBC and MIC effects on Escherichia coli bacteria and Tanacetum polycephalum had the least effects on the bacteria, as well. For the most part, the species of S. verticillata had the robustest essences against the bacteria. Table 2. The MIC and MBC rates (mg/ml) of the herbs essences on the bacteria Bacteria Gram S. aureus E. coli Positive Negative S. lavandulifolia MIC MBC 4.3 8 2.15 4.3 S. verticillata MIC MBC 0.30 0.61 1.23 2.47 T. polycephalum MIC MBC 1 4 8 16 202 Intl J Farm & Alli Sci. Vol., 4 (3): 197-206, 2015 C) The effects of essences on bacteria using diffusion method in agar substrate The most antibacterial effects on E. coli bacteria were found from the essential oils of S. verticillata species while the essences of S. verticillata and T. polycephalum had the most antibacterial effects on S. aureus in comparison with Streptomycin antibiotic (Fig. 1). a) Staphylococcus aureus b) Escherichia coli Figure 1. The antibacterial effects of the essential oils (EO) of S. lavandulifolia (S.L), S. verticillata (S.V), and T. polycephalum (T.P) on lineage in contrast with Streptomycin (ST) antibiotic In light of the bacteria resisting to antibiotics, it is necessary to find new natural products which have antibacterial effects (Lee , 2002) and it is stated that the products from plants are economic and harmless (Lis-Balchin and Deans, 1997; Rostagno and Prado, 2013). As it is reported, Camphor was the main elements of the species of Salvia genus (Chalchat Michet and Pasquier, 1998; Pharmacopoeia, 1983; Weyerstahl , 1999) while this element was not found in the S. verticillata species. (Bayrak and Akgul, 1987) and (Demirci , 2002) had reported that 50% of the essential oil in Salvia fruticosa and S. aramiensis had been formed by 1,8 cineole while Trans-Caryophyllene (18.82%) was the most elements in the S. verticillata which had the effective antibacterial trait on E. coli. The α-thujone and 1-8 cineole were the main elements of the essential oils in planted Salvia officinalis and the α-thujone, 1-8 cineole, camphor, and β-caryophyllenewere the most composition of the essence in S. triloba (Longaray Delamare , 2007). (Strijack , 2004) also reported that the thujone and 1,8 cineole were the most elements in the essential oils of S. fruticosa and the essence had considerably effects on E. coli. The achieved results have precisely showed that monoterpene compounds were the most elements in the essential oils of the studied herbs which have copious applications for the food and medicinal industries. (Tucker, , 1996) have reported that the most elements in the essential oils of T. fruticulosum were the monoterpene hydrocarbons, as well. (Askin Akpulat , 2005) and (Bagci and Kocak, 2010) have also claimed different components in the Tanacetum genus. The species of S. verticillata had the most antibacterial effects on Staphylococcus aureus and S. lavandulifolia had the least one on same bacteria. The gram-positive bacteria of S. aureus has rigid peptidoglycan walls which can resist against many essential effects while the gram-negative bacteria of E. coli has frail phospholipid walls which can be fragile against the essence effects, as well. Hence, the S. aureus was the most resistible bacterial in this study. The essential oils of T. polycephalum also had the most antibacterial effects on S. aureus in the second ranking. The most antibacterial effects of the essences and their components refer to hydrophobia traits in the essential oils which are caused to infiltrate the lipidic walls of bacterial cells and can be changed its structures (Skaltsa , 2003) and consequently many ions and cellular substrates percolate toward to outside of cells and be ultimately dying of bacteria (Carson, Mee and Riley, 2002). It is reported that the resistance of microbial cells depend on solvation rate of antibacterial matters in the lipidic portion of cellular membrane (Lanciotti , 2004). Therefore, differences of antibacterial effects of various species’ essential oils can be justified as the cellular structures. The research of (Skaltsa , 1999) has been showed that Tanacetum parthenium had the robust MIC and MBC on some bacteria. (Saha , 2013) have also reported same capability against gram- positive and negative bacteria from the Tanacetum polycephalum species. Furthermore, other researchers have been reported that different herbal essences had considerable inhibition effects on microorganisms which were contamination agents in food matters (Burt, 2004; Mehdizadeh and Razavi’Rouhani, 2009). Other researchers have also reported the antibacterial effects of different species of Salvia genus (Bouaziz , 2009, Vagionas 2007, Vjera Bilusic Vundac , 2006). Plentiful 203 Intl J Farm & Alli Sci. Vol., 4 (3): 197-206, 2015 researches have been done on antimicrobial effects of diverse essences, extractions, and condiments that show highly abilities of these sorts of components to prevent the growth of pathogenic microorganisms and decomposing of food matters (Ababutain 2011). Regarding this, the study on species of Stachys genus showed that sesquiterpene formed the most parts of all species essence (Chalchat , 2000; Flamini , 2005; Tepe , 2005; Weyerstahl, 1999). (Maleki , 2001) clarified the main and whole components of Stachys lavandulifolia and some its antibacterial effects. Some studies also showed the antibacterial effects of Stachys genus regarding its essential oils’ compounds (Dulger , 2005; Tavassoli, 2011). Conclusion Regarding the results, it can be stated that using of essential oils of the plants species are more reliable and economic approaches to control and repel the bacterial activities. The herb of S. verticillata has the most antibacterial effects on the different, gram- positive or negative, bacteria although its essential rate, as S. lavandulifolia, is so less than the T. polycephalum. On the other hand, sometimes the least amount of essence has the most affection as it is precisely showed in this study. Hence, in order to protect the food products against many bacteria and their resistance to many synthetic antibiotics, it is suggested that the natural products of plants, e.g. their essences, can be used to force back the microbial effects. REFERENCES Ababutain IM. 2011. Antimicrobial activity of ethanolic extracts from some medicinal plant, Australian Journal of Basic and Applied Sciences, 5(11): 678-683. Adams RP. 2007. Identification of Essential Oil Components by Gas Chromatography/Mass Spectrometry, 4th Edition. Allured Publishing Corporation. USA, 804 p. Akhondzadeh’Basti A, Misaghi A, Moosavi M, Zahraei’Salehi M and Gittii K. 2007. The effects of essential oils of Zataria multiflora on growth rate of Staphylococcus aureus in commercial soups. Iranian Research journal of Aromatic and Medicinal Plants, (22): 91-98. (In Persian) Albuquerque UP, Monteiro JM, Ramos MA and Amorim ELC. 2007. Medicinal and magic plants from a public market in Northeastern Brazil. J. Ethnopharmacol, 110:76-91. Askin Akpulat H, Tepe B, Sokmen A, Daferera D and Polissiou M. 2005. Composition of the essential oils of Tanacetum argyrophyllum (C. Koch) Tvzel. var. argyrophyllum and Tanacetum parthenium (L.) Schultz Bip. (Asteraceae) from Turkey Original Research Article. Biochemical Systematics and Ecology, 33(5):511-516. Babakhanlo P, Mirzai M, Sefidkon F, Ahmadi L, Barazandeh MM and Asgari F. 1998. (Flor of Iran.Medical and aromatic plant research instate of forests and rangelands.1st ed.Tehran. Ministry of Jahan-e-Agriculture, 64-82. (In Persian) Bagci E and Kocak A. 2010. Essential oil composition of two endemic Tanacetum (T. nitens (Boiss.&Noe) Grierson and T. argenteum (Lam.) Willd. subsp. argenteum) (Asteraceae) taxa, growing wild in Turkey, Industrial Crops and Products, 31(3):542-545. Barazandeh M. 2003. Quantitative and qualitative investigation of essential oil of Tanacetum polycefalum [J]. Iran J Med Aromat Plants, 19(2):111116. Barbosa WLR, do Nascimento MS, do Nascimento Pinto L, Maia FLC, Sousa AJA, Júnior JOCS, Monteiro MM and de Oliveira DR. 2012. Selecting Medicinal Plants for Development of Phytomedicine and Use in Primary Health Care, Bioactive Compounds in Phytomedicine, Prof. Iraj Rasooli (Ed.), ISBN: 978-953-307-805-2, InTech, Available from:http://www.intechopen.com/books/bioactive-compounds-inphytomedicine/selecting medicinal-plants-for-development-of phytomedicine -and-use-in-primary-health-care. Baron E and Finegold S. 1990. Diagnostic Microbiology. Mosby Co., USA, 861p. Bayrak A and Akgul A. 1987. Composition of essential oils from Turkish Salvia species Phytochemistry, 26: 846-847. Beuchat LR. 2001. Control of Foodborne Pathogens and Spoilage Microoganisms by Naturally Occurring Antimicrobials, Microbiaial Food Contamination, CRC Press, London. Bouaziz M, Yangui T, Sayadi S and Dhouib A. 2009. Disinfectant properties of essential oils from Salvia officinalis L. cultivated in Tunisia. Food and Chemical Toxicology, 47: 2755-2760. Burt S. 2004. Essential oils: their antibacterial properties and potential applications in foods- a review. InternationalJournal of Food Microbiology, 94: 223-253. Carson CF, Mee BJ and Riley TV. 2002. Mechanism of action of Melaleuca alternifolia (tea tree) oil on Staphylococcus aureus determined by time-kill, lysis, leakage and salt tolerance assays and electron microscopy. Antimicrobial Agents and Chemotherapy, 46: 1914-1920. Çetin B, Çakmakçi S and Çakmakçi R. 2011. The investigation of antimicrobial activity of thyme and oregano essential oils. Turk J Agric For., 35:145-154. Chalchat JC, Michet A and Pasquier B. 1998. Study of clones of Salvia officinalis L. yields and chemical composition of essential oil. Flavour and Fragrance Journal, 13:68-70. Chalchat JC, Petrovoic SD, Maksimovic ZA and Gorunovic MS. 2000. Composition of Essential Oils of Salvia nemorosa L. and Salvia glutinosa L., Arh. farm., 3-4: 310-311. Chamorro ER, Zambón SN, Morales WG, Sequeira AF and Velasco GA. 2012. Study of the Chemical Composition of Essential Oils by Gas Chromatography, Gas Chromatography in Plant Science, Wine Technology, Toxicology and Some Specific Applications, Dr. Bekir Salih (Ed.), ISBN: 978-953-51-0127-7, InTech, Available from: http://www.intechopen.com/books/gas-chromatography-in-plant-science-winetechnology-toxicology-and-some-specific-applications/study-of-the-chemical-composition-of-essential-oils-by-gas-chromatography. Demirci B, Baser KHC and Tumen G. 2002. Composition of the essential oil of Salvia Composition of the essential oil of Salvia aramiensis Rech. Fil. growing in Turkey, Flavour and Fragrance Journal, 17: 23-25. Dulger B, Ugurlu E, Aki C, Suerdem TB and Camdevrin A. 2005. Evaluation of antimicrobial activity of some endemic Scrophulariaceae members from Turkey. Pharmaceutical Biology, 43(3):275-279. 204 Intl J Farm & Alli Sci. Vol., 4 (3): 197-206, 2015 Fatouma AL, Edou P, Eba F, Mohamed N, Ali A, Djama S, Obame LC, Bassolé I and Dicko M. 2010. Antimicrobial and antioxidant activities of essential oil and methanol extract of Jasminum sambac from Djibouti. African Journal of Plant Science, 4(3): 038-043. Flamini G, Pier LC, Ivano M, Sezgin CR, Suleyman G and Orhan U. 2005. Essential oil of Stachys aleurites from Turkey. Biochemical Systematics and Ecology, 33(1): 61–66. Floros JD, Newsome R and Fisher W. 2010. Feeding the world today and tomorrow: the importance of food science and technology. Comprehensive Reviews in Food Science and Food Safety, 0: 1-28. Doi: 10.1111/j.1541-4337.2010.00127.x Gandomi’Nasrabadi H, Misaghi A, Akhondzadeh’Basti A, Khosravi A, Bokaei S and Abasifar A. 2008. The effects of essential oils of Zataria multiflora on growth rate of Aspergilus flavus. Journal of Medicinal Plants, (27): 45-51. (in Persian) Ghahraman A. 1994. Cormophytes of Iran. Iran University Press, Tehran. (In Persian) Ghahraman A. 1988. Colour Flora of Iran. Press in Research Institute of Forest and Rangelands, V.11, 1352p. (In Persian) Goncalves MJ, Cruz MT, Cavaleiro C, Lopes MC and Salgueiro L. 2010. Chemical, antifungal and cytotoxic evaluation of the essential oil of Thymus zygis subsp. Sylvestris, Industrial Crops and Products, 32: 70–75. Gurib-Fakim A. 2006. Medicinal plants: Traditions of yesterday and drugs of tomorrow. Molecular Aspects of Medicine, 27:1–93. Holley RA and Patel D. 2005. Improvement in Shelf-Life and Safety of Perishable Foods by Plant Essential Oils and Smoke Antimicrobials. Food Microbilogy, 22: 273-292. ITC. 2012. Packaging for organic foods. International Trade Centre (ITC), Switzerland, 83p. Javidnia, K., R. Miri, M.R. Moein, M. Kamalinejad and H. Sarkarzadeh, 2006. Constituents of essential oil of Stachys pilifera benth from Iran. J Essential Oil Res (JEOR), 18: 275-277. Kumar P. 2009. Green healers: a review. Paper including in ‘Medicinal plants Utilisation and Conservation, 2nd Revised and Enlarged Edition edited by P.Ch. Trivedi, Jaipur-302 004, India, 542p’. Lanciotti R, Gianatti A, patrignani F, belletti N, Guerzoni ME and Gardini F. 2004. Use of natural aroma compounds to improve shelf life and safety of minimally processed fruits. J. food Science & Technology, 15: 201-208. Lee YL, Cesario T, Yang W, Edward S, Hanbrom M and Lauri T. 2002. Antibacterial activity of vegetables and juices Plant. Journal of, Cell and Environment, 25: 1031-1037. Lis-Balchin M and Deans SG. 1997. Bioactivity of selected plant essential oils against Listeria monocytogenes. J. Appl. Bacteriol, 82:759–762. Longaray Delamare AP, Moschen-Pistorello IT, Artico L, Atti-Serafini L and Echeverrigaray S. 2007. Antibacterial activity of the essential oils of Salvia officinalis L. and Salvia triloba L. cultivated in South Brazil. Chemistry, 100: 603-608. Maleki N, Garjani A, Nazemiyeh H, Nilfouroshan Eftekhar AS and Allameh H. 2001. Potent anti-inflamatory activities of hydroalcholic extract from aerial parts of Stachys inflate on rat. J. Ethnopharmacol, 75: 13-218. Marilena C, Bersani C and Comi G. 2001. Impedance measurements to study the antimicrobial activity of essential oils from Lamiaceae and Compositae. International Journal of Food Microbiology, 67: 187–195. Marriout PJ, Shellie R and Cornewell C. 2001. Gas chromatographic technologies for the analysis of essential oils. Journal of Chromatography A, 935: 1-22. Mehdizadeh T and Razavi’Rouhani SM. 2009. The antibacterial effects of the essential oils of three onions on staphylococcus aureus bacteria. Journal of Agricultural and Natural Resources, 15(2):122-134. (In Persian) Monteiro JM, de Souza JSN, Lins Neto EMF, Scopel K and Trindade EF. 2014. Does total tannin content explain the use value of spontaneous medicinal plants from the Brazilian semi-arid region? Rev Bras Farmacogn, 24: 116-123. Mortazavi A. 2009. Applied Microbiology of food and in vitro. Ferdowsi University press, 594p. Mos’hafi MH, Mofidi A, Mehrbani M and Mehrbani M. 2009. Investigation of factures and anti-bacterial effects of essential oil in Stachys acerosa Boiss. Medicinal plants journal, 9(33): 108-115. Mothana RA, Al-Said MS, Al-Yahya MA, Al-Rehaily AJ and Khaled JM. 2013. GC and GC/MS Analysis of Essential Oil Composition of the Endemic Soqotraen Leucas virgata Balf.f. and Its Antimicrobial and Antioxidant Activities. Int J Mol Sci., 14(11): 23129-23139. Mukherjee T. 2009. Medicinal Plants: Need for Protection. Paper including in ‘Medicinal plants Utilisation and Conservation, 2nd Revised and Enlarged Edition edited by P.Ch. Trivedi, Jaipur-302 004, India, 542p’. Mukherjee PK, Maity N, Nema NK and Sarkar BK. 2011. Bioactive compounds from natural resources against skin aging. Phytomedicine, 19:6473. Ndawonde BG. 2006. Medicinal Plant Sales: A Case Study in Northern Zululand, Ms. Thesis, University of Zulnland, 127p. Nori-Shargh D, Norouzi D, Mirza M, Jaimand K and Mohammadi S. 1999. Chemical composition of the essential oil of Tanacetum polycephalum (SchulTz Bip. SSP Heterophyllum). Flavour and Fragrance Journal, 14: 105-106. Ozkan G, Sagdic O, Gokturk S, Unal O and Albayrak S. 2010. Study on chemical composition and biological activities of essential oil and extract from Salvia pisidica. LWT - Food Science and Technology, 43(1): 186-190. Pauwels E, Stoven V and Yamanishi Y. 2011. Predicting drug side-effect profiles: a chemical fragment-based approach. BMC Bioinformatics, 12:169-182. Pharmacopoeia E. 1983. Vol 1. Maissoneuve, SA: Sainte Ruffine, 4392p. Rabbani M, Saggadi SE and Zarei HR. 2003. Anxiolytic effects of Stachys lavandulifolia vahl on the elevated plus–maze model of anxiety in mice. J. Ethnopharmacol, 89: 6 -271 Rabbani M, Saggadi SE and Jalali A. 2005. Hydroalcohol extractandfractions of Stachys lavandulifolia vahl: effect on spontaneous motor activity and elevated plus–maze behaviour. Phytother Res, 19: 8-854. Rasooli I. 2012. Bioactive Compounds in Phytomedicine. InTech publication, Croatia, 228p. Rechinger KH and Hedge IC. 1982. Flora Iranica Akademiche Druck verlag sanstalt, Gras, Austria, PP, 150-159: 360 – 365. Reynolds J. 1996. Martindale–the Extra Pharmacopoeia, London. Royal Pharmaceutical Society of Great Britain, 31:120-129. Rezvanpanah S, Rezaei K, Golmakani MT and Razavi SH. 2011. Antibacterial properties and chemical characterization of the essential oils from summer savoury extracted by microwave-assisted hydrodistillation. Brazilian Journal of Microbiology, 42: 1453-1462. Rostagno MA and Prado JM. 2013. Natural Product Extraction, Principles and Applications. Published by The Royal Society of Chemistry, UK, 516p. Saha S, Nath Dhar T, Sengupta C and Ghosh P. 2013. Biological Activities of Essential Oils and Methanol Extracts of Five Ocimum Species against Pathogenic Bacteria. Czech J. Food Sci., 31(2):194-202. Saharkhiz M, Sattari M, Goodarzi GH and Omidbeygi R. 2008. Determining the effect of anti-bacterial essential of Tanacetum parthenium, J. Sci. Res. Iran Aromat. Herbs, 24(1): 47-55. (In Persian) 205 Intl J Farm & Alli Sci. Vol., 4 (3): 197-206, 2015 Sikkema J, De Bont JAM and Poolman B. 1994. Intractions of cyclic hydrocarbons with biological membranes. Journal of Biological Chemistry, 269: 8022-8028. Sivropolulov A, Skokini T and Arsenakas M. 1995. Antimicrobial activity of mint essential oil, j. Agric food chem, 43: 2334-2388. Skaltsa HD, Lazari DM, Chinou IB and Loukis AE. 1999. Composition and antibacterial activity of the essential oils of Stachys candida and S. chrysantha from southern Greece. Planta Med, 65: 255-256. Skaltsa HD, Demetzos C, Lazari D and Sokovic M. 2003. Essential oil analysis and antimicrobial activity of eight Stachys species from Greece. Phytochemistry, 64 (3):743-752. Strijack C, Harding GKM, Ariano RE and Zelenitsky S. 2004. Peritoneal Fluid Titer Test for Peritoneal Dialysis-Related Peritonitis. Antimicrobial Agents Chemotherapy, 48(5): 1719-1726. Tavassoli S, Mousavi SM, Emam-Djomeh Z and Razavi SH. 2011. Comparative Study of the Antimicrobial Activity of Rosmarinus officinalis L. Essential Oil and Methanolic Extract. Middle-East Journal of Scientific Research, 9(4): 467-471. Tepe B, Donmez E, Unlu M, Candan F, Daferera D, Vardar-Unlu G, Polissiou M and Sokmen A. 2004. Antimicrobial and antioxidative activities of essential oils and methanol extracts of Salvia cryptantha (Montbret et Aucher ex Benth.) and Salvia multicaulis (Vahl.). Food Chemistry, 84:519-525. Tepe B, Daferera D, Sokmen A, Sokmen M and Polissiou M. 2005. Antimicrobial and antioxidant activities of the essential oil and various extracts of Salvia tomentosa Miller (Lamiaceae). Food Chemistry, 3(90):333-340. Tucker AO, Maciarello MJ and Clebsch BB. 1996. Volatile leaf oil of Salvia clevelandii (Gray) Greene Gilman. Journal of Essential Oil Research, 8: 669-670. Vagionas K, Graikou K, Ngassapa O, Runyoro D and Chinou I. 2007. Composition and antimicrobial activity of the essential oils of three Satureja species growing in Tanzania. Food Chemistry, 103: 319–324. Vjera Bilusic Vundac HW, Pfeifhofer AH, Brantner ZM and Misko P. 2006. Essential oils of seven Stachys taxa from Croatia. Biochemical Systematics and Ecology, 34(12): 875-881. Weyerstahl P, Marschall H, Thefeld K and Rustaiyan A. 1999. Constituents of the essential oil of Tanacetum (syn Chrysanthemum) fruticulosum Ledeb, from Iran. Flavour and Fragrance Journal, 14:112-120. 206