Untitled Document

CrossMark

Original Article

Bull Emerg Trauma 2018;6(4):355-362.

Antimicrobial Survey of Local Herbal Drugs against Acinetobacter baumannii Isolated from Patients Admitted to a Level-I Trauma Center

Hossein Abdolrahimzadeh1, Shahram Bolandparvaz1, Hamid Reza Abbasi1, Maryam Dehghankhalili2, Shahram Paydar1*, AmirReza Dehghanian1, Salar Hafez Ghoran3,4, Mojtaba Asadollahi3, Mehdi Zare3

1Trauma Research Center, Shahid Rajaee (Emtiaz) Trauma Hospital, Shiraz University of Medical Sciences, Shiraz, Iran
2Student Research Committee, Department of General Surgery, Shiraz University of Medical Sciences, Shiraz, Iran
3Medicinal and Natural Products Chemistry Research Center (MNCRC), Shiraz University of Medical Sciences, Shiraz, Iran
4Faculty of Basic Sciences, Department of Chemistry, Golestan University, Gorgan, Iran

.........................

*Corresponding Author: Shahram Paydar
Address: Trauma Research Center, Shahid Rajaee (Emtiaz) Trauma Hospital, Shiraz University of Medical Sciences, Shiraz, Iran. Tel/Fax: +98-71-36254206 ,e-mail: paydarsh@gmail.com

Received: May 13, 2018
Revised: July 22, 2018
Accepted: July 29, 2018

........................

Please cite this paper as: 
Abdolrahimzadeh H, Bolandparvaz S, Abbasi HR, Dehghankhalili M, Paydar S, Dehghanian AR, Hafez Ghoran S, Asadollahi M, Zare M. Antimicrobial Survey of Local Herbal Drugs against Acinetobacter baumannii Isolated from Patients Admitted to a Level-I Trauma Center. Bull Emerg Trauma. 2018;6(4):355-362. doi: 10.29252/beat-060414.

.........................

Abstract

Objective: To determine the antimicrobial activity and entity of several local herbal plants against Acintobacters isolated from trauma patients admitted to a Level-I trauma center.
Methods: The antibacterial activities of the Satureja bachtiarica oil and someselected Iranian medicinal plants (Artemisia sieberi and Tanacetum dumosum belonging to the Asteraceae/Compositae; Salvia mirzayanii and Mentha mozaffarianii belonging to the Lamiaceae/Labiatae) were assayed on A. baumannii by microdilution and agar disc diffusion methods. Having obtained the acceptable antibacterial data, the shade-dried aerial parts of the plants were extracted by hydrodistillation method using Clevenger apparatus according to European pharmacopeia for 3 h. The analysis of S. bachtiarica essential oil accompanied by other herbal drug oils were performed by using GC/FID and GC/MS methods.
Results: Outcomes revealed that the S. bachtiarica essential oil exhibited the potent antibacterial capability against Acinetobacter strains in comparison with Colistin, as a positive control. For S. bachtiarica,the growth inhibition zone and minimum inhibitory concentration (MIC) values were 21 mm and 0.5 mg/ml, while, for Colistin, the data were in order: 8 mm and 0.016 mg/ml. Consequently, GC/MS outcomes demonstrated that the major components of the essence were carvacrol (48.6%), followed by p-Cymene (16.6%), γ-terpinene (6.9%) and linalool (5.3%).
Conclusion: Based on the considerable inhibitory activity against nosocomial infections by essential oil of S. bachtiarica, it could be considered as the suitable candidate in the food industry and pharmaceutical uses.

Keywords: Trauma; Acintobacter baumannii; Antibacterial activity;Satureja bachtiarica; Carvacrol.

.........................

Introduction
Trauma is a cellular disruption caused by an environmental energy that is beyond the body's resilience. Trauma remains the most common cause of death for all individuals between the ages of 1 and 44 years and is the third most common cause of death regardless of age. It is also the number one cause of years of productive life lost. Death after trauma has a trimodal distribution. Acute deaths (less than 24 hours) usually result from uncontrolled conditions, but infections and multiple organ dysfunction syndrome, which often arise from infection, are responsible for a significant proportion of late deaths. Indeed, infection is responsible for most deaths in patients who survive longer than 48 hours after trauma. Trauma infections can be divided into the injury and nosocomial infections. Most post traumatic infections are polymicrobial, involving a mixture of aerobic and anaerobic organisms. In one series, 37-45% of all trauma patients experienced infectious complications during their initial hospitalization. In that same study, 80% of all trauma patients who were in ICU at least 7 days met the criteria for SIRS [1]. Humans have evolved mechanisms to avoid infection despite presence of bacteria in our environment. Under normal circumstances there is a balance between our microbial, intact environmental barriers, and host defenses. Traumatic injury disrupts this balance and significantly increases the probability of developing an infection [1].
Acinetobacter baumannii is an aerobic Gram-negative bacillus which is non-fermentative, catalase-positive and oxidase-negative [2]. Considering the resistance to multiple antimicrobial agents along with the wide spreading nosocomial infections, A. baumannii has been evolved from being an underestimated microorganism to one of the most important factors causing the various infections at hospital environments, mainly in immunocompromised patients over the last 30 years [3-6]. Surprisingly, the intrinsic resistance of Acinetobacters could be related to not only the genetic materials including plasmids, transposons and integrons but also the ability to survive on anhydrous surfaces for the long interval periods [3,7-9]. Multidrug-resistant A. baumannii strains are frequently found to be responsible for epidemics of nosocomial infections, such as respiratory, bloodstream, urinary tract, skin, and soft tissue infections [10].
According to ethnopharmacological survey in Iranian folk medicine, we found some medical plants that used to prevent post-operative infection rate [11,12]. Therefore, we evaluated the antibacterial activity of some of these medical plants against Acinetobacters infection. The main purpose of this in vitro study was to evaluate the antibacterial activity of volatile oils obtained from some medicinal plants belonging to the Asteraceae/Compositae (Artemisia sieberi and Tanacetum dumosum) and Lamiaceae (Salvia mirzayanii, Mentha mozaffarianii and Satureja bachtiarica) on Acintobacters isolated from Trauma patients in Shahid Rajaee Hospital of Shiraz.

Materials and Methods
Plant material
The aerial parts of the plants were collected from South and Southwest of Iran in March until July 2017 in the flowering stage of the plants. The plants were identified by regional floras and authors with floristic and taxonomic references [13], and voucher specimens were deposited at the herbarium of Medicinal and Natural Products Chemistry Research Center, Shiraz University of Medical Sciences, Shiraz, Iran. The shade-dried aerial parts of the plants were extracted by hydrodistillation for 3 h, giving relatively high yield oils in all collections (Table 1). The oils were dried over anhydrous sodium sulfate and kept at -4ºC.

Table 1. Chemical constituents (%) of the essential oil of S. bakhtiarica analyzed by GC-FID.

No.

Compounds

Structures

S. bakhtiarica (%)

RI

1

α-Thujene

1.3 ± 0.04

932

2

α-Pinene

1.14 ± 0.08

938

3

Camphene

0.46 ± 0.05

953

4

β-Myrcene

1.71 ± 0.07

993

5

p-Cymene

16.55 ± 1.4

1025

6

Sylvestrene

0.78 ± 00

1031

7

γ-Terpinene

6.86 ± 1

1064

8

cis-Sabinene hydrate

0.47 ± 00

1072

9

m-Cymene

0.3 ± 0.00

1087

10

Linalool

5.25 ± 0.08

1099

11

Borneol

1.85 ± 1

1168

12

Terpinen-4-ol

1.98 ± 0.09

1180

13

Thymol

0.77 ± 00

1291

14

Carvacrol

48.57 ± 2.5

1301

15

Carvacrol acetate

1.96 ± 02

1372

16

β-Caryophyllene

2.08 ± 0.4

1422

17

Spathulenol

0.56 ± 0.02

1550

18

Caryophyllene oxide

1.96 ± 0.4

1589

*The values are the means of three different FID area percentage ± SE; **The major compounds with more than 1.0% area concentration were formatted in bold font.

Gas Chromatography Analysis
Gas Chromatography was performed on a Varian CP-3800 chromatograph, with a FID and a HP-5 column (30 m × 0.25 mm i.d., 0.25 μm film thickness). The oven temperature was programmed from 60°C to 240°C at 5°C/min and finally kept 10 min at this temperature. The carrier gas was N2 with a flow rate of 0.9 mL/min. Injector and detector temperature were set at 250°C. The injection volume was 0.4 μl for the volatile oils and also n-alkanes for calculation of the retention indices was separately injected.

Gas Chromatography-Mass Spectroscopy Analyses
The GC-MS was carried out on an Agilent 7890N chromatograph, coupled to an Agilent 5975C mass spectrometer (Agilent Technologies, USA), operating at 70 eV ionization energy, 0.5 s/scan and the mass range: 35-400, equipped with a HP-5 MS capillary column (30 m × 0.25 mm, film thickness 0.25 μm). Helium was used as the carrier gas at the constant flow of 0.9 ml/min and split ratio of 1:50. The oven temperature was 60°C rising to 250°C at a rate of 5°C /min, then held at 250°C for 10 min; transfer line temperature was set at 250°C.

Identification of compounds
Retention indices (RI) for all constituents were calculated according to Van den Dool approach [14], using n-alkanes (C6 – C22) as standards and the essential oils on a HP-5 column under the same chromatographic conditions. The identification of the components was made based on comparison of their mass spectra with those of the internal computer reference mass spectra libraries (Wiley 7.0 and Nist), as well as by comparison of their retention indices with data mentioned in the previous literatures [15].

Antibacterial Activity
The antimicrobial activity of the obtained essential oils was tested by using the disc-diffusion method on Muller Hinton Agar (MHA) [16] using determination of inhibition zones, and determining the minimal inhibitory concentration (MIC) using the macro dilution broth susceptibility assay as recommended by NCCLS [17].

Study Population and Sampling
All trauma patients admitted in Shahid Rajaee Hospital, the unique center of trauma in Shiraz, affiliated to Shiraz University of Medical Sciences (SUMS) hospital with single of sepsis and positive blood culture for Acintobacter baumannii. The hospital is a referral center of trauma for Fars Province situated in Southern part of Iran.

Microorganisms and Media
Disc diffusion assay:
Muller Hinton agar (MHA) was autoclaved at 121°C for 20 min. The plates (8-cm diameter) were prepared with 10 ml agar inoculated with 1 ml of each bacterial suspension. Microorganisms were cultured for 16 to 24 h at 37°C and prepared to turbidity equivalent to McFarland Standard No. 0.5. Sterile paper discs (6 mm in diameter) were impregnated with 20 μl of dilutions of known essential oil concentrations (5 μg/disc) and incubated at 37°C for 24 h. The essential oils were dissolved in dimethyl sulfoxid (DMSO, 15 μl) before the test for antimicrobial activity. Discs (6 mm diameter) of Colistin (10 μg) was used as positive controls. Bacterial growth inhibition was determined as the diameter of the inhibition zones around the discs (mm). The growth inhibition diameter was an average of three measurements, taken at three different directions. All tests were performed in triplicate.

Minimum Inhibitory Concentration (MIC) Using Muller Hinton Broth Micro-Dilution (MHB)
The minimal inhibitory concentration (MIC) values were evaluated using the broth serial dilution method according to standard methods [18]. The MIC was defined as the lowest concentration of the compound to inhibit the growth 50% of microorganisms. Bacterial strains were cultured overnight at 37 °C in Muller Hinton Broth (MHB, Oxoid). Stock solution of the essential oil was prepared in 33.3% (v/v) dimethyl sulfoxide (DMSO). Dilution series, using MHB, were prepared from 0.062-8 mg/ml. After incubation at 37 °C for 24 h, the microorganism growth inhibition was evaluated by measuring absorbance at 600 nm, and prepared to turbidity equivalent to McFarland Standard No. 0.5 using a spectrophotometer. An aliquot of the samples (5 μl) was added to 95 μl of fresh media followed by 100 μl of the bacterial suspension (OD=0.1 at 600 nm) in a 96-well plate. The plates were incubated at 37ºC for 24 h in a shaking incubator. Colistin was used as positive control in each assay. DMSO solution was used as a negative control. Control tubes were incubated under the same condition. Antibacterial activity was detected using a colorimetric method by adding 10 μl of 0.5% INT solution in water in each well at the end of the incubation period for further 30 min. Experiments were performed in triplicate but at three different times.

Results
In terms of measuring the antibacterial activity using disc diffusion method, the significant zones of inhibition around the discs were noted in Tables 2 and 3. Our examinations investigated the antimicrobial activities of medicinal plants belonging to the Asteraceae (Artemisia sieberi and Tanacetum dumosum) and Lamiaceae (Salvia mirzayanii, Mentha mozaffarianii and Satureja bachtiarica) families. Careful results on examined bacterial strains revealed that the essential oil of Satureja bachtiarica Bunge assayed by both Disk Diffusion and Minimum Inhibitory Concentration (MIC) methods was the most potent one against all of Acintobacters (Figures 1 and 2). Our findings showed that the aerial part of S. bachtiarica essential oil had high ability against Gram-negative Acintobacter strains. The essential oil seems to be active against all bacterial strains used (MIC values of 0.5 mg/ml). Hence, the most active concentration was 0.5 mg/ml inhibiting completely the growth of all the clinically isolated Acintobacters.  
The hydrodistillation of S. bachtiarica aerial parts led to a yellowish oil in 0.6% (w/w) yield. Then, the obtainedvolatile oil was analyzed by GC-FID and GC-MS. Therefore, the main constituents were identified as the oxygenated monoterpenes: carvacrol (48.57%) and linalool (5.25%); monoterpene hydrocarbons; p-cymene (16.55%) and γ-terpinene (6.86%) (Table 1) corresponding to 94.55% of the essential oil.

Table 2. Antimicrobial potential (MICa) of essential oil from S. bachtiarica, against 6 Acintobacters isolated from individual Trauma patients in Shahid rajaei Hospital of Shiraz, Iran as determined by nutrient-broth micro-dilution bioassay.

 

Acintobacter

Plant oil

From Ttauma patient 1

From Ttauma patient 2

From Ttauma patient 3

From Ttauma patient 4

From Ttauma patient 5

From Ttauma patient 6

Satureja bachtiarica

0.5

0.5

0.5

0.5

0.5

0.5

Colistin

0.016

0.016

0.016

0.016

0.016

0.016

 

 

 

*Minimum inhibitory concentration (MIC) the plant extracts in the bacterial suspension in the nutrient broth media (mg/ ml) determined in three replicates

Table 3. Disk diffusion results of five medical plants against 6 Acintobacters isolated from Trauma patients in Shahid rajaei Hospital of Shiraz, Iran.

 

Acintobacter

Plant oil

From Ttauma patient 1

From Ttauma patient 2

From Ttauma patient 3

From Ttauma patient 4

From Ttauma patient 5

From Ttauma patient 6

Mentha mozaffarianii

NAa

NAa

NAa

NAa

NAa

NAa

Artemisia sieberi

NAa

NAa

NAa

NAa

NAa

NAa

Salvia mirzayanii

NAa

NAa

NAa

NAa

NAa

NAa

Tanacetum dumosum

NAa

NAa

NAa

NAa

NAa

NAa

Satureja bachtiarica

21 mm

20 mm

21 mm

19 mm

21 mm

21 mm

Colistin

8 mm

8 mm

8 mm

8 mm

8 mm

8 mm

 

 

 

*Concentration of essential oil all medical plants is 2.5 mg/ml; aNA= not active

 

Fig. 1. Antimicrobial potential (MICa) of essential oil from S. bachtiarica, against Acintobacters. isolated from Trauma patients in Shahid rajaee Hospital of Shiraz, Iran as determined by nutrient-broth micro-dilution bioassay.

Fig. 2. Disk diffusion bioassay of five medical plants against Acintobacter mentioned in below.

 

Discussion
Although the antibacterial activity and chemical constituents of the essential oil of S. bachtiarica have been previously reported [12,19,20], in agreement with the literature, the major constituents of the oil were characterized as carvacrol, p-cymene and thymol. Interestingly, our data revealed that the other major components after carvacrol and p-cymene were γ-terpinene and linalool. The Satureja species is commonly used as a spice and traditionally as a muscle pain reliever, tonic, and carminative in treating stomach and intestinal disorders such as cramps, nausea, indigestion, and diarrhea [21].
According to the phenyl backbone, the remarkable antimicrobial activity of S. bachtiarica may be because of the presence of mentioned compounds. It is suggested that these components could synergistically inhibited the various bacterial strains. Therefore, the good antimicrobial activity of carvacrol is certainly justified by the hypothesis proposed by A. Ben Arfa et al., [22] with related to its hydrophobic behavior in the membrane along with detoxifying the unwanted free radicals. The importance of the hydroxyl group in the phenolic structure was confirmed in terms of activity when carvacrol was compared to its methyl ether. Furthermore, the relative position of the hydroxyl group exerted an influence upon the components effectiveness as seen in the difference in activity between carvacrol and thymol against Gram-negative and Gram-positive bacteria. However, the significance of the phenolic ring was demonstrated by the lack of activity of the monoterpene cyclic hydrocarbon p-cymene. The high activity of the phenolic components may be further explained in terms of the alkyl substitution into the phenol nucleus, which is known to enhance the antimicrobial activity of phenols [23]. These compounds were strongly active despite their relatively low capacity to dissolve in water, which is in agreement with published data [24-35]. In another research, the antimicrobial activity of carvacrol was examined on various Gram-positive and Gram-negative bacterial strains. Results revealed that carvacrol was significantly inhibited all selected bacteria at different concentrations (400 and 800 μg/ml). The obtained data showed maximum inhibition against Serratia spp. (28 mm), followed by E. coli (26 mm) and Enterobacter spp. (25 mm), Klebsiella pneumonia (23 mm), Proteus mirabilis (22 mm), S. aureus (20 mm), S. epidermidis (22 mm) and St. pneumonia (16 mm), surprisingly, there are no effect on Pseudomonas aeroginosa [36]. It seems to be that the function of carvacrol as a phenolic essential oil could be due to the following manners including to disintegrate the outer membrane of bacterial cells, to be the disturbance of the cytoplasmic membrane, disrupting the proton motive force (PMF), electron flow, active transport and coagulation of cell contents [37,38]. Likewise, the potency of carvacrol in disintegrating the cell membrane of gram negative bacteria were confirmed by means of enhancing lipopolysaccharides (LPS) accompanied by the increasing permeability of the cytoplasmic membrane to ATP [39]. On the basis of a comprehensive survey on essential oils isolated from various species of Satureja, some biological properties such as anti-bacterial, anti-fungal, anti-viral [40-42], anti-oxidant [43], antispasmodic, anti-diarrheal, anti-nociceptive, anti-inflammatory [44,45], anti-HIV-1 [40] and immunomodulatory effects [45] have been previously mentioned. S. bachtiarica known as Marzeh-e-Koohi is an Iranian endemic specie and aromatic plant that commonly observed among mountain in the southwestern part of Iran [46]. In the folk medicine, the plant is used as analgesic and antiseptic by Bakhtiari and Chaharmahali tribes of Iran [47]. The major phenyl backbone components of S. bachtiarica were carvacrol, thymol and p-cymene [48].
According to the considerable antibacterial activity of essential oils noted in the literatures and some problematic features existed by some bacterial strains, paying attention to explore the natural drugs from nature in order to repair various human defeats is worthwhile.
In conclusion, the S. bachtiarica (known as “Marzeh” in Persian)essential oil clearly demonstrates antibacterial properties, although the mechanistic processes are poorly understood. These activities suggest potential use as chemotherapeutic agents, food preserving agents and disinfectants.

Acknowledgments: The authors are grateful to Shahid Rajaee (Emtiaz) Trauma Hospital, Medicinal and Natural Products Chemistry Research Center (MNCRC) and Shiraz University of Medical Sciences for financial and kind support.
Conflict of interest: All authors declare there is no conflict of interest.

..........................

References

  1. Morales CH, Villegas MI, Villavicencio R, Gonzalez G, Perez LF, Pena AM, et al. Intra-abdominal infection in patients with abdominal trauma. Arch Surg. 2004;139(12):1278-85; discussion 85.
  2. Vila J, Pachon J. Therapeutic options for Acinetobacter baumannii infections. Expert Opin Pharmacother. 2008;9(4):587-99.
  3. Bergogne-Bérézin E, Friedman H, Bendinelli M. Acinetobacter: Biology and pathogenesis. Springer Science & Business Media; 2008.
  4. Gordon NC, Wareham DW. Multidrug-resistant Acinetobacter baumannii: mechanisms of virulence and resistance. Int J Antimicrob Agents. 2010;35(3):219-26.
  5. Falagas ME, Karageorgopoulos DE. Pandrug resistance (PDR), extensive drug resistance (XDR), and multidrug resistance (MDR) among Gram-negative bacilli: need for international harmonization in terminology. Clin Infect Dis. 2008;46(7):1121-2; author reply 2.
  6. Pachon J, Vila J. Treatment of multiresistant Acinetobacter baumannii infections. Curr Opin Investig Drugs. 2009;10(2):150-6.
  7. Higgins PG, Dammhayn C, Hackel M, Seifert H. Global spread of carbapenem-resistant Acinetobacter baumannii. J Antimicrob Chemother. 2010;65(2):233-8.
  8. Mugnier PD, Poirel L, Naas T, Nordmann P. Worldwide dissemination of the blaOXA-23 carbapenemase gene of Acinetobacter baumannii. Emerg Infect Dis. 2010;16(1):35-40.
  9. Vila J, Marti S, Sanchez-Cespedes J. Porins, efflux pumps and multidrug resistance in Acinetobacter baumannii. J Antimicrob Chemother. 2007;59(6):1210-5.
  10. Seifert H, Dijkshoorn L, Gerner-Smidt P, Pelzer N, Tjernberg I, Vaneechoutte M. Distribution of Acinetobacter species on human skin: comparison of phenotypic and genotypic identification methods. J Clin Microbiol. 1997;35(11):2819-25. 
  11. Khaledi M, Asadi-Samani M, Mahmoodi-Kouhi A, Gholipour A. Antibacterial Effect of The Hydroalcoholic Extracts of Four Iranian Medicinal Plants on Staphylococcus aureus and Acinetobacter baumanii. International Journal of Pharmaceutical And Phytopharmacological Research. 2017;7(2):10-4.
  12. Pirbalouti AG, Malekpoor F, Enteshari S, Yousefi M, Momtaz H, Hamedi B. Antibacterial activity of some folklore medicinal plants used by Bakhtiari tribal in Southwest Iran. International Journal of Biology. 2010;2(2):55.
  13. Cantino P, Harley R, Wagstaff S. Genera of Labiatae status and classification. Royal Botanica Gardens Kew; 1992.
  14. Vandendool H, Kratz PD. A generalization of the retention index system including linear temperature programmed gas-liquid partition chromatography. J Chromatogr. 1963;11:463-71.
  15. Adams RP. Identification of essential oil components by gas chromatography/quadrupole mass spectroscopy. USA: Allured Publishing Corporation; 2001.
  16. Wikins TD, Holdeman LV, Abramson IJ, Moore WE. Standardized single-disc method for antibiotic susceptibility testing of anaerobic bacteria. Antimicrob Agents Chemother. 1972;1(6):451-9.
  17. Waitz JA. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically. National Committee for Clinical Laboratory Standards; 1990.
  18. Slavkovska V, Jancic R, Bojovic S, Milosavljevic S, Djokovic D. Variability of essential oils of Satureja montana L. and Satureja kitaibelii wierzb. ex Heuff. from the central part of the balkan peninsula. Phytochemistry. 2001;57(1):71-6.
  19. Sefidkon F, Sadeghzadeh L, Teimouri M, Asgari F, Ahmadi Sh. Antimicrobial effects of the essential oils of two Satureja species (S. Khuzistanica Jamzad and S. bachtiarica Bunge) in two harvesting time. Iran J Medic Arom Plants. 2007;23:174-82.
  20. Teimori M. Essential oil analysis and antibacterial activity of Satureja bachtiarica Bunge in Ardebile province. J Plant Sci Res. 2009;14:19-26.
  21. Zargari A. Medicinal Plants. 4th ed. Tehran: University of Tehran Publication; 1990. p. 42-45.
  22. Ben Arfa A, Combes S, Preziosi-Belloy L, Gontard N, Chalier P. Antimicrobial activity of carvacrol related to its chemical structure. Lett Appl Microbiol. 2006;43(2):149-54.
  23. Pelczar M, Chan E, Krieg N. Control of microorganisms, the control of microorganisms by physical agents. Microbiology. 1988;469:509.
  24. Nadal NM, Montalvo A, Seda M. Antimicrobial properties of bay and other phenolic essential oils. Cosmetics and Perfumery. 1973;88:37-8.
  25. Suresh P, Ingle V, Vijayalakshmi V. Antibacterial activity of eugenol in comparison with other antibiotics. Journal of food science and technology. 1992;29(4):254-6.
  26. Lattaoui N, Tantaoui-Elaraki A. Individual and combined antibacterial activity of the main components of three thyme essential oils. Rivista Italiana EPPOS. 1994;13:13-9.
  27. Mahmoud AL. Antifungal action and antiaflatoxigenic properties of some essential oil constituents. Letters in Applied Microbiology. 1994;19(2):110-3.
  28. Meena MR, Sethi V. Antimicrobial activity of the essential oils from spices. J Food Sci Tech Mysore. 1994;31:68-70.
  29. Shapiro S, Meier A, Guggenheim B. The antimicrobial activity of essential oils and essential oil components towards oral bacteria. Oral Microbiol Immunol. 1994;9(4):202-8.
  30. Belaiche T, Tantaoui-Eleraki A, IbrahimY A. Application of a two levels factorial design to the study of the antimicrobial activity of three terpenes. Sciences des Aliments. 1995;15(6):571-8.
  31. Hsouna AB, Trigui M, Mansour RB, Jarraya RM, Damak M, Jaoua S. Chemical composition, cytotoxicity effect and antimicrobial activity of Ceratonia siliqua essential oil with preservative effects against Listeria inoculated in minced beef meat. Int J Food Microbiol. 2011;148(1):66-72.
  32. Charai M, Mosaddak M, Faid M. Chemical composition and antimicrobial activities of two aromatic plants: Origanum majorana L. and O. compactum Benth. Journal of Essential Oil Research. 1996;8(6):657-64.
  33. Sivropoulou A, Papanikolaou E, Nikolaou C, Kokkini S, Lanaras T, Arsenakis M. Antimicrobial and cytotoxic activities of Origanum essential oils. Journal of agricultural and Food Chemistry. 1996;44(5):1202-5.
  34. Hili P, Evans CS, Veness RG. Antimicrobial action of essential oils: the effect of dimethylsulphoxide on the activity of cinnamon oil. Lett Appl Microbiol. 1997;24(4):269-75.
  35. Lis-Balchin M, Deans SG. Bioactivity of selected plant essential oils against Listeria monocytogenes. J Appl Microbiol. 1997;82(6):759-62.
  36. Bnyan I, Abid A, Obied H. Antibacterial activity of carvacrol against different types of bacteria. Journal of Natural Sciences Research. 2014;4(9):13-7.
  37. Davidson PM, Taylor TM, Schmidt SE. Chemical preservatives and natural antimicrobial compounds.  Food microbiology: American Society of Microbiology; 2013. p. 765-801.
  38. Lambert RJ, Skandamis PN, Coote PJ, Nychas GJ. A study of the minimum inhibitory concentration and mode of action of oregano essential oil, thymol and carvacrol. J Appl Microbiol. 2001;91(3):453-62.
  39. Helander IM, Alakomi H-L, Latva-Kala K, Mattila-Sandholm T, Pol I, Smid EJ, et al. Characterization of the action of selected essential oil components on Gram-negative bacteria. Journal of agricultural and food chemistry. 1998;46(9):3590-5.
  40. Yamasaki K, Nakano M, Kawahata T, Mori H, Otake T, Ueba N, et al. Anti-HIV-1 activity of herbs in Labiatae. Biol Pharm Bull. 1998;21(8):829-33.
  41. Sokovic M, Tzakou O, Pitarokili D, Couladis M. Antifungal activities of selected aromatic plants growing wild in Greece. Nahrung. 2002;46(5):317-20.
  42. Abad MJ, Bermejo P, Gonzales E, Iglesias I, Irurzun A, Carrasco L. Antiviral activity of Bolivian plant extracts. Gen Pharmacol. 1999;32(4):499-503.
  43. Radonic A, Milos M. Chemical composition and in vitro evaluation of antioxidant effect of free volatile compounds from Satureja montana L. Free Radic Res. 2003;37(6):673-9.
  44. Hajhashemi V, Sadraei H, Ghannadi AR, Mohseni M. Antispasmodic and anti-diarrhoeal effect of Satureja hortensis L. essential oil. J Ethnopharmacol. 2000;71(1-2):187-92.
  45. Ghasemi Pirbalouti A, Pirali E, Pishkar G, Jalali SM, Reyesi M, Jafarian Dehkordi M, et al. The essential oils of some medicinal plants on the immune system and growth of rainbow trout (Oncorhynchus mykiss). Journal of Herbal Drugs (An International Journal on Medicinal Herbs). 2011;2(2):149-55.
  46. Jamzad Z. A new species of the genus Satureja (Labiatae) from Iran. Iran J Bot. 1994;6(2):215-8. (in Persian).
  47. Pirbaloutl A. Medicinal plants used in Chaharmahal and Bakhtyari districts of Iran. Herba Polonica. 2009;55(2):69-77.
  48. Sefidkon F, Jamzad Z. Essential oil of Satureja bachtiarica Bunge. Journal of Essential Oil Research. 2000;12(5):545-6.

PDF     XML   FIGS

Journal compilation © 2018 Trauma Research Center, Shiraz University of Medical Sciences

Open Access License

All articles published by Bulletin of Emergency And Trauma are fully open access: immediately freely available to read, download and share. Bulletin of Emergency And Trauma articles are published under a Creative Commons license. Mandated authors will be offered CC-BY; all other authors will choose between CC-BY, CC-BY-NC and CC-BY-NC-ND.