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