MICROBIAL DRUG RESISTANCE

Volume 18, Number 2, 2012

ª Mary Ann Liebert, Inc.
DOI: 10.1089/mdr.2011.0080

The Antimicrobial Activity of Thyme Essential Oil Against

Multidrug Resistant Clinical Bacterial Strains

Monika Sienkiewicz,1 Monika Łysakowska,1 Paweł Denys,2 and Edward Kowalczyk 3

Aim: The aim of this work was to investigate the antimicrobial activity of thyme essential oil against clinical
multidrug resistant strains of Staphylococcus, Enterococcus, Escherichia, and Pseudomonas genus. Materials:
The antibacterial activity of oil was tested against standard strains of bacteria and 120 clinical strains iso-
lated from patients with infections of the oral cavity, abdominal cavity, respiratory and genitourinary tracts,
skin, and from the hospital environment. Methods: Agar diffusion was used to determine the microbial
growth inhibition of bacterial growth at various concentrations of oil from Thymus vulgaris. Susceptibility
testing to antibiotics was carried out using disk diffusion. Results: Thyme essential oil strongly inhibited the
growth of the clinical strains of bacteria tested. Conclusions: The use of phytopharmaceuticals based on an
investigated essential oil from thyme in the prevention and treatment of various human infections may
be reasonable.

Introduction The aim of the study

O pportunistic infections caused by Gram-negative

bacteria, mainly of the Klebsiella sp., Enterobacter sp.,
Serratia sp., Proteus sp., Escherichia sp., and Pseudomonas sp.
present rising problems.55 There has also been a significant
increase in the number of infections caused by Gram-positive
cocci belonging to Staphylococcus sp. and Enterococcus
sp.11,20,30 Thus, the search for effective and safe medicines
that could be used to treat particularly persistent bacterial
infections is on. Oils, a diverse group of plant metabolites,
seem to be an interesting solution; in past years, a large
number of essential oils and their constituents have been
investigated for their antimicrobial properties against certain
bacteria, fungi, viruses, and protozoa.
The essential oils of thyme, oregano, mint, cinnamon, cu-
min, salvia, clove, and eucalyptus have been found to pos-
sess the strongest antimicrobial properties.33 Many of them
appear to have a wide spectrum of antibiotic activity against
microflora that provokes hospital-acquired infections. The
broad and complex activity of essential oils, as well as their
synergy of action, can make them a valued weapon against
multidrug resistant bacterial strains. What is more, there is
no evidence of emergence of resistant bacteria after their
usage, and this is highly promising in the treatment of hu-
man diseases.25,34,43,49
The aim of this work was to investigate the antimicrobial
properties of thyme essential oil obtained from thyme (Thy-
mus vulgaris L.) against standard and clinical strains of bac-
teria isolated from patients, clinical staff, and the hospital
environment.
Materials and Methods
Bacterial strains
The standard strains, Staphylococcus aureus ATCC 433000,
Enterococcus faecalis Van B ATCC 51299, En. faecalis, vanco-
mycin VA-sensitive ATCC 29212, Enterococcus faecium, VA-
sensitive ATCC 35667, Enterococcus durans, VA-sensitive
ATCC 6656, Escherichia coli ATCC 25922, and Pseudomonas
aeruginosa ATCC 27853 came from the collection of the
Medical and Sanitary Microbiology Department, Medical
University of Lodz.
The clinical strains of Staphylococcus, Enterococcus, Escher-
ichia, and Pseudomonas genera were collected from different
materials originating from patients and medical staff, as well
as the environment of various wards from two hospitals in
Lodz: internal diseases, surgery, urology, laryngology, and
intensive care unit. Bacterial strains were isolated from ab-
dominal cavity exudates (n = 4), bronchial secretions (n = 5),

1
Medical and Sanitary Microbiology Department, Medical University of Lodz, Medical University of Lodz, Lodz, Poland.
2
Orthopedic Clinic, Medical University of Lodz, Orthopedic Clinic, Lodz, Poland.
3
Pharmacology and Toxicology Department, Medical University of Lodz, Medical University of Lodz, Lodz, Poland.

137
138
nose (n = 9), ear (n = 3), pharynx (n = 6), hands (n = 2), ulcers
(n = 14), wounds (n = 22), bedsores (n = 12), abscesses (n = 1),
groin (n = 5), bile (n = 1), toe (n = 1), anus (n = 6), sputum
(n = 1), blood (n = 2), urine (n = 14), drains (n = 2), hospital
staff (n = 2), disinfecting dispensers (n = 2), hospital beds
(n = 2), and cupboard swabs (n = 4).
Bacterial strain identification
S. aureus strains were identified to the genus according to
standard microbiological methods: culturing on Columbia
Agar (bioMerieux) with 5% blood, culturing on Mannitol Salt
Agar (bioMerieux), using Slidex-Staph Kit (bioMerieux), and
determining the ability of bacteria to produce catalase and
coagulase (bioMerieux). Microorganisms were identified to
the species by using API 20 Staph tests (bioMerieux). Bacteria
were incubated at 37C for 24 hours. S. aureus ATCC 29213
strain was used as a control.
Enterococci were identified to the genus according to
standard microbiological methods: culturing on Columbia
Agar (bioMerieux) with 5% sheep blood, culturing on En-
terococcosel Agar (Emapol), determining the presence of D
antigen according to Lancefield (Slidex-Strep Kit, bioMer-
ieux), and determining the ability of bacteria to produce
catalase, pyrrolidonyloarylamidase (Lachema). The micro-
organisms were identified to the species by using API 20
Strep tests (bioMerieux). The bacteria were incubated at 37C
for 24 hours. As a control, En. faecalis Van B ATCC 51299, En.
faecalis ATCC 29212, En. faecium ATCC 35667, and En. durans
ATCC 6656 strains were used.
Es. coli strains were identified according to standard mi-
crobiological methods such as culturing on Columbia Agar
(bioMerieux) with 5% blood and culturing on Mac Conkey
Agar (bioMerieux) and were identified to the species by us-
ing API 20 E tests (bioMerieux). Bacteria were incubated in
37C for 24 hours. As a control, Es. coli ATCC 25922 strain
was used.
P. aeruginosa strains were identified according to standard
microbiological methods including culturing on Agar and
Columbia Agar (bioMerieux) with 5% blood, culturing on
Mac Conkey Agar (bioMerieux), and determining the ability
of the bacteria to produce oxidase (bioMerieux). The micro-
organisms were identified in the species by using API 20 NE
tests (bioMerieux). The bacteria were incubated in 37C for 24
hours. As a control, P. aeruginosa ATCC 27853 strain was used.
Essential oil analysis
Commercial essential oil from thyme—T. vulgaris L. (La-
miaceae) was purchased from the manufacturer (POLLENA-
AROMA) and analyzed by GC-FID-MS in the Institute of
General Food Chemistry, Technical University of Lodz, us-
ing a Trace GC Ultra apparatus (Thermo Electron Corpora-
tion) MS DSQ II detectors and FID-MS splitter (SGE).
Operating conditions: apolar capillary column Rtx-1 ms
(Restek), 60 m · 0.25 mm i.d., film thickness 0.25 mm; tem-
perature program, 50C–300C at 4C/minute; SSL injector
temperature 280C; FID temperature 300C; split ratio 1:20;
carrier gas helium at a regular pressure 200 kPa.; FID tem-
perature 260C; carrier gas, helium; 0.5 ml/min; split ratio
1:20. Mass spectra were acquired over the mass range of 30–
400 Da, ionization voltage of 70 eV, and ion source temper-
ature of 200C.
SIENKIEWICZ ET AL.
Identification of components was based on the compari-
son of their MS spectra with those of the laboratory-made
MS library, commercial libraries (NIST 98.1, Wiley Registry
of Mass Spectral Data, 8th Ed. and MassFinder 3.1) and with
literature data1,31 along with the retention indices on apolar
column (Rtx-1, MassFinder 3.1) associated with a series of
alkanes with linear interpolation (C8-C26). A quantitative
analysis (expressed as percentages of each component) was
carried out by peak area normalization measurements
without correction factors.
The standard and clinical strains were cultivated in Co-
lumbia agar medium and incubated at 37C for 48 hours in
aerobic conditions. Bacterial suspensions with an optical
density of 0.5 on the Mc Farland scale were prepared and
analyzed with a Bio Merieux densitometer.
The antibacterial properties of the tested oil were investi-
gated by agar dilution. The essential oil was diluted in eth-
anol. This solution was mixed with a nutrient broth to obtain
concentrations from 0.03125 to 2.5 ml/ml and poured into
petri dishes. Inoculum containing 1.5$108 CFU (0.1 ml) per
spot was seeded on the surface of the agar with various oil
concentrations, as well as on agar with no oil added (strains
growth control). Minimal inhibitory concentration (MIC)
was determined after 24 hours of incubation at 37C in aer-
obic conditions. The analysis of the antibacterial activity of
the oil was performed thrice independently.
Susceptibility testing
The following antibiotics (Becton Dickinson) were used for
susceptibility testing of S. aureus strains: cefoxitin (FOX;
30 mg) (R £ 14, 15 £ I £ 17, S ‡ 18), erythromycin (E; 15 mg)
(R £ 13, 14 £ I £ 22, S ‡ 23), clindamycin (CC; 2 mg) (R £ 14,
15 £ I £ 20, S £ 21), nitrofurantoin (F/M; 300 mg) (R £ 14,
15 £ I £ 16, S ‡ 17) (for isolates from urine), VA (30 mg) (S ‡ 15),
teicoplanin (TEC; 30 mg) (R £ 10, 11 £ I £ 13, S ‡ 14), tetracy-
cline (TE; 30 mg) (R £ 14, 15 £ I £ 18, S ‡ 19), chloramphenicol
(C; 30 mg) (R £ 12, 13 £ I £ 17, S ‡ 18), ciprofloxacin (CIP; 5 mg)
(R £ 15, 16 £ I £ 20, S ‡ 21), trimethoprim/sulfamethoxazole
(SXT; 1.25 mg/23.75 mg) (R £ 10, 11 £ I £ 15, S £ 16), fusidic acid
(FA; 10 mg) (S ‡ 22), linezolid (LZD; 30 mg) (S ‡ 21); of En-
terococcus genus:
Ampicillin (AM; 10 mg) (R £ 16 S ‡ 17), C (30 mg) (R £ 12,
13 £ I £ 17, S ‡ 18), CIP (5 mg) (R £ 15, 16 £ I £ 20, S ‡ 21), E
(15 mg) (R £ 13, 14 £ I £ 22, S £ I £ 23), fosfomycin (FOS; 200 mg)
(R £ 12, 13 £ I £ 15, S ‡ 16) (only for En. faecalis, the isolates
from urine), F/M (300 mg) (R £ 14, 15 £ I £ 16, S ‡ 17) (for iso-
lates from urine), gentamicin (GM; 120 mg) (R £ 6, 7 £ I £ 9,
S ‡ 10), LZD (30 mg) (R £ 20, 21 £ I £ 22, S ‡ 23), imipenem
(IPM; 10 mg) (R £ 13, 14 £ I £ 15, S ‡ 16), penicillin (P; 10 mg)
(R £ 14, S ‡ 15), streptomycin (S; 300 mg) (R £ 6, 7 £ I £ 9, S ‡ 10),
synercid (SYN; 4.5 mg/10.5 mg) (R £ 15, 16 £ I £ 18, S ‡ 19) (only
for En. faecium), TE (30 mg) (R £ 14, 15 £ I £ 18, S ‡ 19), TEC
(30 mg) (R £ 10, 11 £ I £ 13, S ‡ 14), VA (30 mg) (R £ 14,
15 £ I £ 16, S ‡ 17), of Es. coli strains: amoxicillin/clavulanic
acid (AMC; 20 mg/10 mg) (R £ 13, 14 £ I £ 17, S ‡ 18), cefalotin
(CF; 30 mg) (R £ 14, 15 £ I £ 17, S ‡ 18), cefazolin (CZ; 30 mg)
(R £ 14, 15 £ I £ 17, S ‡ 18), cefuroxime (CXM; 30 mg) (R £ 14,
15 £ I £ 17, S ‡ 18), GM (10 mg) (R £ 12, 13 £ I £ 14, S ‡ 15), AM
(10 mg) (R £ 13, 14 £ I £ 16, S ‡ 17) (only for the isolates from
urine), norfloxacin (NOR; 10 mg) (R £ 12, 13 £ I £ 16, S ‡ 17) (as
above), F/M (300 mg) (R £ 14, 15 £ I £ 16, S ‡ 17) (as above),
THE ANTIMICROBIAL ACTIVITY OF THYME ESSENTIAL OIL 139

FOX (30 mg) (R £ 14, 15 £ I £ 17, S ‡ 18), cefotaxim (CTX; 30 mg) Table 1. Components of the Essential Oil Obtained
(R £ 14, 15 £ I £ 22, S ‡ 23), ceftazidime (CAZ; 30 mg) (R £ 14, from Thyme—Thymus vulgaris L. (Lamiaceae)
15 £ I £ 17, S ‡ 18), aztreonam (ATM; 30 mg) (R £ 15, 16 £ I £ 21,
Total Retention
S ‡ 22), IPM (10 mg) (R £ 13, 14 £ I £ 15, S ‡ 16), CIP (5 mg)
No. Compound oil % indices
(R £ 15, 16 £ I £ 20, S ‡ 21), netilmicin (NET; 30 mg) (R £ 12,
13 £ I £ 14, S ‡ 15), tobramycin (NN; 10 mg) (R £ 12, 13 £ I £ 14, 1 a-Thujene 0.6 932
S ‡ 15), C (30 mg) (R £ 12, 13 £ I £ 17, S ‡ 18), TE (30 mg) (R £ 14, 2 a-Pinene 1.9 936
15 £ I £ 18, S ‡ 19), SXT (1.25 mg/23.75 mg) (R £ 10, 11 £ I £ 15, 3 Camphene 1.2 950
S ‡ 16) and of P. aeruginosa strains: mezlocillin (MZ; 75 mg) 4 Oct-1-en-3-ol 1.0 962
(R £ 17, 18 £ I £ 20, S ‡ 21), piperacillin (PIP; 100 mg) (R £ 17, 5 b-Pinene 0.3 978
18 £ I £ 20, S ‡ 21), CAZ (30 mg) (R £ 14, 15 £ I £ 17, S ‡ 18), GM 6 Myrecene 1.1 987
(10 mg) (R £ 12, 13 £ I £ 14, S ‡ 15), NN (10 mg) (R £ 12, 7 p-Cymene 29.1 1,015
8 1.8-Cineole 2.1 1,024
13 £ I £ 14, S ‡ 15), AMC (20 mg/10 mg) (R £ 13, 14 £ I £ 17,
9 Limonene 0.2 1,025
S ‡ 18), piperacillin/tazobactam (TZP; 100 mg/10 mg) (R £ 17, 10 g-Terpinene 5.2 1,051
18 £ I £ 20, S ‡ 21), CTX (30 mg) (R £ 14, 15 £ I £ 22, S ‡ 23), ATM 11 p-Cymenene 0.1 1,075
(30 mg) (R £ 15, 16 £ I £ 21, S ‡ 22), IPM (10 mg) (R £ 13, 12 Terpinolene 0.1 1,082
14 £ I £ 15, S ‡ 16), meropenem (MEM; 10 mg) (R £ 13, 13 Linalool 3.7 1,086
14 £ I £ 15, S ‡ 16), NET (30 mg) (R £ 12, 13 £ I £ 14, S ‡ 15), CIP 14 Camphor 0.5 1,123
(5 mg) (R £ 15, 16 £ I £ 20, S ‡ 21), SXT (1.25 mg/23.75 mg) 15 Borneol 1.9 1,150
(R £ 10, 11 £ I £ 15, S ‡ 16), C (30 mg) (R £ 12, 13 £ I £ 17, S ‡ 18), 16 Terpinen-4-ol 1.3 1,164
and colistin (CL; 50 mg) (R < 15, S > 15). Susceptibility testing 17 a-Terpineol 0.3 1,176
was carried out using the disk-diffusion method, on Mueller- 18 Thymol methyl ether 1.3 1,215
19 Carvacrol methyl ether 1.0 1,226
Hinton II Agar. Cultures were incubated at 37C for 16–18
20 Borneol acetate 0.3 1,270
hours, VA for 24 hours. The results were interpreted ac- 21 Thymol 38.1 1,267
cording to Clinical and Laboratory Standard Institute 22 Carvacrol 2.3 1,278
guidelines.9 23 Thymol acetate 0.2 1,329
24 African-1-en 0.1 1,356
Results 25 a-Copaene 0.2 1,379
26 b-Burbonene 0.1 1,386
Chemical composition of the tested oil 27 b-Caryophyllene 3.1 1,421
The composition of the essential oil derived from T. vul- 28 Thymohydroquinone 0.1 1,509
garis was found to meet the requirements of the Polish 29 a-Humulene 0.1 1,455
30 g-Muurolene 0.3 1,474
Pharmacopoeia VIII and the European Pharmacopoeia.18,40
31 cis-b Guaiene 0.1 1,488
The content of thymol amounts to 38.1% and carvacrol 32 Cuparene 0.1 1,498
to 2.3%, as well as other prevailing compounds such as 33 g-Cadinene 0.6 1,507
p-cymene (29.1%), g-terpinene (5.2%), and linalool (3.7%). The 34 Calamenene B 0.2 1,517
chemical composition of the tested oil is shown in Table 1. 35 d Cadinene 0.3 1,520
36 a-Cadinene 0.1 1,534
Susceptibility testing 37 Caryophyllene oxide 0.5 1,578
38 g-Eudesmol 0.1 1,618
The tested strains of S. aureus were resistant to 50% of 39 Eudesm-3-en-7-ol 0.1 1,650
b-lactams and macrolides recommended for susceptibility 40 Cadalene 0.1 1,659
testing. Clinical strains of the Enterococcus genus showed
resistance to macrolides in almost 70%, to b-lactams and Bold texts and values indicate the major constituents of thyme oil.
aminoglycosides in 60%, and to carbapenems in 40%. The
examined Es. coli strains were resistant to b-lactams in 40%
The tested clinical strains of S. aureus, resistant to many
and to aminoglycosides in 20%. The tested P. aeruginosa
antibiotics, were sensitive to the thyme oil at low concen-
strains were resistant to b-lactams in more than 50%, to
trations. S. aureus strain isolated from abscesses was resistant
carbapenems in 30%, and to aminoglycosides and mono-
to 7 out of 11 tested antibiotics (sensitive to VA, TEC, C, and
bactams in 20%. The results are shown in Tables 2–5.
LZD); the MIC value for the thyme oil was 0.25 ml/ml. The
MIC for highly multidrug resistant bacterial strains from the
The activity of thyme oil against tested bacterial strains
hand, wound, ear, foot ulceration, exudates from episiotomy
The MIC for S. aureus were between 0.25 and 1.0 ml/ml. wounds, stump ulcers, and urine ranged from 0.5 ml/ml to
MIC was 0.25 ml/ml for the standard strain of S. aureus 0.75 ml/ml. Thyme oil at 0.5 ml/ml concentration inhibited the
ATCC 433000 and 6 strains from the clinical material. The growth of more than 45% of the resistant strains. Table 2
results are shown in Figure 1. shows the general characteristics of S. aureus isolates.
Most Staphylococcus strains were sensitive to oil at a con- The VA resistant standard strain—En. faecalis Van B ATCC
centration of 0.5 ml/ml. Growth inhibition was obtained in 17 51299 was the most sensitive strain to thyme essential oil
out of the 30 clinical bacterial strains. Isolates were taken (MIC–0.0625 ml/ml). The MIC was 0.75 ml/ml for VA sensi-
from the nose (n = 5), ear (n = 1), hand (n = 2), wound (n = 3), tive strains of En. faecalis ATCC 29212 and En. faecium ATCC
abdominal cavity (n = 1), groin (n = 1), ulcer swabs (n = 3), and 35667 and MIC for the standard strain of En. durans ATCC
urine (n = 1). The results are presented in Figure 2. 6656 was 0.85 ml/ml. The results are shown in Figure 1.
140 SIENKIEWICZ ET AL.

Table 2. Characteristics of Staphylococcus aureus Isolates

Susceptibility to antibacterials Total

Staphylococcus aureus MIC of thyme
No. strain/clinical material essential oil ll/ml FOX E CC F/M VA TEC TE C CIP SXT FA LZD R I S

1. Swab/nose 0.25 R S R — S S R S I S S S 3 1 7
2. Swab/nose 0.5 S S S — S S R S S S S S 1 — 10
3. Swab/nose 0.75 R S R — S S R S I R R S 5 1 5
4. Swab/nose 0.5 R S R — S S R S I I R S 4 2 5
5. Swab/nose 0.5 R S R — S S R S I S R S 4 1 6
6. Swab/nose 0.25 S S S — S S S S S R R S 2 — 9
7. Swab/nose 0.5 S S S — S S R S S R S S 2 — 9
8. Swab/nose 0.5 S S S — S S S S S I S S — 1 10
9. Swab/ear 0.5 R R R — S S R S R R R S 7 — 4
10. Swab/hend 0.5 S S S — S S R S S S S S 1 — 10
11. Swab/hend 0.5 R R R — S S R S R R R S 7 — 4
12. Swab/wound 0.75 R S R — S S R S I R R R 6 1 4
13. Swab/wound 0.75 R S R — S S R S I R S S 4 1 6
14. Swab/wound 0.5 S S S — S S R S R S S S 2 — 9
15. Swab/wound 0.5 I R R — S S R R S S S S 4 1 6
16. Swab/wound 0.5 S S S — S S R S S S S S 1 — 10
17. Swab/wound 1.0 R R R — S S R S R S R S 6 — 5
18. Exudation/abdominal cavity 0.25 R R R — S S R S R S S S 5 — 6
19. Exudation/abdominal cavity 0.5 I R R — S S R S R S S S 4 1 6
20. Exudation/abdominal cavity 0.25 S R R — S S R S R S S S 4 — 7
21. Swab/ulceration 0.5 R R R — S S R S R I R S 6 1 4
22. Swab/ulceration 0.5 S R R — S S R S R S S S 4 — 7
23. Swab/ulceration 0.75 R R R — S S S R R R R S 7 — 4
24. Swab/ulceration 0.5 S S S — S S S S S S S S — — 11
25. Swab/groin 0.5 S S S — S S S S S S S S — — 11
26. Swab/groin 1.0 I R R — S S R R I S S S 4 2 5
27. Swab/abscess 0.25 R R R — S S R S R R R S 7 — 4
28. Urine 0.5 I R R S S S R S R S S R 5 1 6
29. Urine 0.25 R S R S S S R S I R S S 4 1 7
30. Swab/drain 0.75 S S S — S S R S S S S S 1 — 10

FOX, cefoxitin; E, erythromycin; CC, clindamycin; F/M, nitrofurantoin; VA, vancomycin; TEC, teicoplanin; TE, tetracycline; C,
chloramphenicol; CIP, ciprofloxacin; SXT, trimethoprim/sulfamethoxazole; FA, fusidic acid; LZD, linezolid; R, resistant; I, intermediate
susceptible strain; S, susceptible strain; MIC, minimal inhibitory concentration.

Thyme oil was also very active against clinical strains, the
most active against En. faecium strain being isolated from the
ulcer (MIC–0.25 ml/ml); it significantly inhibited 16 clinical
strains at a concentration of 0.75 ml/ml and 11 clinical strains
at 0.5 ml/ml. The results are shown in Figure 2.
An MIC value of - 0.5 ml/ml was characteristic for all
strains of En. faecalis and En. faecium isolated from urine, and
En. faecalis strains derived from the hospital environment.
The concentration of 0.75 ml/ml inhibited the growth of
En. faecalis clinical strains from wounds and the throat, as
well as of En. faecium strains from blood and from the hos-
pital environment.
It was found that most clinical resistant strains of entero-
cocci were sensitive to the tested oil. The En. faecium strain
isolated from ulcers, resistant to 9 out of 14 antibiotics
(sensitive to LNZ, SYN, TEC, and VA), was sensitive to
thyme essential oil at the lowest concentration of 0.25 ml/ml.
All En. faecium clinical strains isolated from urine, resistant to
8 antibiotics, were also sensitive to the oil of thyme; MIC was
0.5 ml/ml. In all, more than 45% of resistant strains were
sensitive to thyme oil: MIC = (0.75 ml/ml). Table 3 shows the
general characteristics of Enterococcus sp. isolates.
The oil showed bactericidal activity against Es. coli ATCC
25922 standard strain at 0.25 ml/ml (Fig. 1). Figure 2 shows
that the same MIC value was determined for 13 strains from
the clinical material. Thyme oil of this concentration was
effective against clinical strains isolated from the throat
and the majority of strains from the wound swabs. An MIC
of - 0.5 ml/ml was obtained for 17 tested clinical strains of
colon bacilli, isolated from the anus, bedsore swabs, and
urine. The clinical strains of Es. coli tested that were highly
resistant to commonly used antibiotics showed sensitivity
to thyme oil.
The multidrug resistant strain isolated from ulcers, re-
sistant to 14 out of 16 antibiotics (sensitive to the ATM,
IPM), was sensitive to thyme oil (MIC = (0.25 ml/ml)). The
same MIC was obtained for two strains from wound and
bedsore swabs, which were sensitive to only 5 out of the 16
tested antibiotics. The growth of nearly 60% of the resistant
strains was inhibited by thyme oil at a concentration of
0.5 ml/ml. Table 4 shows the general features of Es. coli
isolates.
Strains of P. aeruginosa were the most resistant to the
thyme oil. The standard strain—P. aeruginosa ATCC 27853—
was inhibited with 0.5 ml/ml. The results are shown in Figure
1. The number of the clinical strains that were sensitive to the
thyme oil at 1.5, 2.0, and 2.5 ml/ml was similar, and it is
presented in Figure 2. Concentrations of 1.5 and 2.0 ml/ml
THE ANTIMICROBIAL ACTIVITY OF THYME ESSENTIAL OIL 141

Table 3. Characteristics of Enterococcus sp. Isolates

MIC of thyme Susceptibility to antibacterials Total
Enterococcus spp. essential
No. Strain/clinical material oil ll/ml AM C CIP E FOS F/M GM IPM LNZ P S SYN TE TEC VA R I S

1. En. faecalis/urine 0.5 S S R S S S S S S S S — I S S 1 1 12

2. En. faecalis/urine 0.5 R S R R S S R R S R R — R S S 8 — 6
3. En. faecalis/urine 0.5 S S I I S S S S S S S — R S S 1 2 11
4. En. faecalis/urine 0.5 S R R R S S R S S R R — R S S 7 — 7
5. En. faecalis/urine 0.5 S R R R S S R S S S R — R S S 6 — 8
6. En. faecalis/swab/wound 0.75 S S I S — — S S S S S — S S S — 1 11
7. En. faecalis/swab/wound 0.75 S R R R — S R S S S R — R S S 6 — 7
8. En. faecalis/swab/wound 0.75 S R R R — — R S S S R — R S S 6 — 6
9. En. faecalis/swab/ulceration 0.75 S R S R — — S S S S I — R S S 3 1 8
10. En. faecalis/bile 0.75 S S S S S S S S S S S — R S S 1 — 13
11. En. faecalis/swab/bedsore 1.25 S S I S — — S S S S S — R S S 1 1 10
12. En. faecalis/swab/pharynx 0.75 S S S I — — S S S S S — R S S 1 1 10
13. E. faecalis/swab/pharynx 0.75 S R I R — — R S S S R — R S S 5 1 6
14. En. faecalis/hospital staff 0.75 S S S R — — S S S S R — R S S 3 — 9
15. En. faecalis/drain 0.75 S S R S — — R S S S S — R S S 3 — 9
16. En. faecalis/cupboard 0.5 S S R S — — R S S S S — R S S 3 — 9
17. En. faecalis/disinfecting dispencer 0.5 S S I S — — S S S S S — R S S 1 1 10
18. En. faecalis/disinfecting dispencer 0.5 S S R S — — R S S S S — R S S 3 — 9
19. En. faecium/urine 0.5 R S R R — S R R S R R S R S S 8 — 6
20. En. faecium/urine 0.5 R S R R — R R R S R R S S S S 8 — 6
21. En. faecium/urine 0.5 R S R R — I R R S R R S R S S 8 1 5
22. En. faecium/blood 0.75 R S R R — — S R S S R S R S S 6 — 7
23. En. faecium/blood 0.75 R S R R — — S R S R R S R S S 7 — 6
24. En. faecium/swab/ulceration 0.25 R I R R — R R R S R R S R S S 9 1 4
25. En. faecium/hospital staff 0.75 R S R R — — S R S R R S R S S 7 — 6
26. En. faecium/hospital bed 0.75 R S R R — — R R S R R S S S S 7 — 6
27. E. faecium/hospital bed 0.75 S S S S — — S S S S S S S S S — — 13
28. En. faecium/cupboard 0.75 R S R R — — R S S S S — R S S 5 — 7
29. En. faecium/cupboard 0.75 R S R R — S R R S R R S R S S 8 — 6
30. En. durans/cupboard 1.0 R S R R — S R R S R R S R S S 8 — 6

AM, ampicillin; FOS, fosfomycin; GM, gentamicin; IPM, imipenem; LZD, linezolid; P, penicillin; S, streptomycin; SYN, synercid.

inhibited the growth of strains mainly isolated from wounds
and bedsores. MIC values of - 2.0 and 2.5 ml/ml were ob-
tained for blue pus bacilli isolated from ulcers. The highest
concentration of thyme oil, 2.5 ml/ml, was effective against
the bacteria from bronchial secretions and from the anus
swabs. It was found that the most resistant micro-organisms
were isolated from bronchial secretions. Two of them were
found to be resistant to all 16 antibiotics, and the third was
sensitive to only 6 of them. The MIC obtained for thyme oil
against these strains ranged from 2.0 to 2.5 ml/ml.
Higher oil activity (MIC-1.5 ml/ml) was obtained against
the P. aeruginosa strain isolated from the flank, which showed
resistance to 12 antibiotics (sensitive to AMC, TZP, IPM, and
NET). At the same time, these strains were sensitive in 40%
of thyme oil used at a concentration of 1.5 ml/ml.
Table 5 shows the general characteristics of P. aeruginosa
isolates.
Control media containing alcohol (at concentrations used
in the dilutions) did not inhibit the growth of bacterial
strains.
Discussion
A number of chemotypes were identified in red thyme,
T. vulgaris; the most important are the thymol (65% thymol,
5%–10% carvacrol) and carvacrol chemotypes (85% carvacrol,
1%–5% thymol).42 According to the requirements of the Polish
Pharmacopeia VIII and European Pharmacopeia, the oil
should contain thymol (36%–55%) and carvacrol (1%–4%).18,40
In our study, the oil showed antimicrobial activity against
standard and clinical strains of S. aureus, En. faecalis,
En. faecium, En. durans, Es. coli, and P. aeruginosa. The ob-
tained results are in accordance with the literature, and show
that thyme oil has strong antimicrobial properties against all
tested strains. The activity is due to the high content of
phenolic compounds with antibacterial properties, such as
thymol and carvacrol, which constitute more than 40% of the
ingredients of the oil.33
In our tests, most of the clinical strains of S. aureus were
sensitive to thyme oil at a concentration of 0.5 ml/ml (17
strains), therefore relatively low compared with the high
concentrations of antibiotics usually required. These strains
came from diverse materials and hospital wards (e.g., swabs
from wounds, swabs from nose, and urine). As far as sus-
ceptibility to antibiotics was concerned, many isolates were
resistant to TE (n = 25, 83.3%), CC (n = 20, 66.6%), FOX (n = 14,
46.6%), E (n = 13, 43.3%), and CIP (n = 12, 40%). However, all
the strains were susceptible to VA, and TEC and only two
(from wound and urine) were resistant to LZD. Strains with
MIC at 1 ml/ml were isolated from swabs (groin and wound),
wards (nephrology and ICU), and remained sensitive to
several antibiotics (VA, TEC, C, SXT, and LNZ). There were
 Table 4. Characteristics of Escherichia coli Isolates

Susceptibility to antibacterials Total

Escherichia coli MIC of thyme
No. strain/clinical material essential oil ll/ml AMC CF CZ CXM GM AM NOR F/M FOX CTX CAZ ATM IPM CIP NET NN C TE SXT R I S

1. Swab/pharynx 0.25 R R R R S — — — S S S S S S S S S I R 5 1 10
2. Swab/pharynx 0.25 R R R R S — — — S S S I S S S S S I R 5 2 9
3. Swab/nose 0.5 S S S S S — — — S S S R S S S R S R S 3 — 13
4. Secretion/bronchical 0.25 R I I S S — — — S R S S S S S S I S S 2 3 11
5. Secretion/bronchical 0.5 R R I S S — — — S R S S S S S S I S S 3 2 11
6. Swab/groin 0.5 R R R R S — — — S S S S S S S S R R R 6 — 9
7. Swab/groin 0.25 R I S S S — — — S S S S S S S S S I S 1 2 13
8. Exudation/abdominal 0.5 R R S S S — — — R R R S S S S R R R R 9 — 7
cavity
9. Swab/anus 0.25 S S S S S — — — S S S S S S S S S S S — — 16
10. Swab/anus 0.5 R I I I S — — — S S S S S S S S S I S 1 4 11
11. Swab/anus 0.5 I R S S S — — — S S S S S S S S S R R 3 1 12
12. Swab/anus 0.5 S S S I S — — — S S S S S S S S I R S 1 2 13
13. Swab/bedsore 0.5 I R R R R — — — S I R S S R R R R R R 11 2 3

142
14. Swab/bedsore 0.5 S I S I S — — — S S I S S S S S S S S — 3 13
15. Swab/bedsore 0.5 R R S S S — — — S S S S S S S S S R S 3 — 13
16. Swab/bedsore 0.5 R R R R S — — — R R R S S R R S S R R 11 — 5
17. Swab/bedsore 0.25 R S S S S — — — S S S S S S S S S R S 2 — 14
18. Swab/ulceration 0.5 R R R R R — — — R R R S S R R R R R R 14 — 2
19. Swab/ulceration 0.25 R R R S S — — — R S S S S S S S S R R 6 — 10
20. Swab/wound 0.25 S S S S S — — — S S S S S S S S S R S 1 — 15
21. Swab/wound 0.5 R R S R S — — — S S R S S S S S S R R 6 — 10
22. Swab/wound 0.5 R R S R S — — — S S S S S S S S R R R 6 — 10
23. Swab/wound 0.25 I I S S S — — — S S S R S S S R S R R 4 2 10
24. Swab/wound 0.25 R S S S S — — — S S S S S S S S S I S 1 1 14
25. Swab/wound 0.25 R R R R R — — — S R S S S R R R S R R 11 — 5
26. Swab/wound 0.25 R R R R R — — — S S S R S R R R S R R 11 — 5
27. Urine 0.5 I R S S S R S S R S S S S S S S S R S 4 1 14
28. Urine 0.5 R R R S S R S S R S S S S S S S R R S 7 — 12
29. Urine 0.25 R R R S S R R S S S S S S R S S S R R 8 — 11
30. Urine 0.5 R R R I S R I S R S S S S I S S I R I 6 5 8

AMC, amoxicillin/clavulanic acid; CF, cefalotin; CZ, cefazolin; CXM, cefuroxime; NOR, norfloxacin; CTX, cefotaxim; CAZ, ceftazidime; ATM, aztreonam; NET, netilmicin; NN, tobramycin.
 Table 5. Characteristics of Pseudomonas aeruginosa Isolates

Susceptibility to antibacterials Total

Pseudomonasaeruginosa MIC of thyme
No. strain/clinical material essential oil ll/ml MZ PIP CAZ GM NN AMC TZP CTX ATM IPM MEM NET CIP SXT C CL R I S

1. Swab/ear 1.0 S S S R S R S S S S S S S S S S 2 — 14
2. Swab/ear 1.0 S S S I S I S S S S S S S S S S — 2 14
3. Swab/pharynx 2.5 R S S S S R S R S S S S I R R S 5 1 10
4. Swab/pharynx 2.0 R S S S S R S R S R S S S R R R 7 — 9
5. Swab/groin 1.5 R R R R R I S R R S R S R R R R 12 1 3
6. Swab/toe 1.5 S S S S S S S R R S R S R R R R 7 — 9
7. Swab/anus 2.5 S S S S S S S R S S S S S R R R 4 — 12
8. Swab/anus 2.5 S S S R R S S R S S S S S R R R 6 — 10
9. Sputum 1.0 S S S S S R S S S S S S S S R I 2 1 13
10. Secretion/bronchical 2.0 R R S I S R S R R R R S R R R S 10 1 5
11. Secretion/bronchical 2.5 R R R R R R R R R R R R R R R R 16 — —
12. Secretion/bronchical 2.5 R R R R R R R R R R R R R R R R 16 — —
13. Swab/bedsore 1.5 R R R S R R S I R S R I I R R R 10 3 3

143
14. Swab/bedsore 1.5 R R S S S R S R S S S R R R R S 8 — 8
15. Swab/bedsore 1.5 R S S S S R S R S S S S S R R S 5 — 11
16. Swab/bedsore 2.0 R S S S S R R R S S S S S R R R 7 — 9
17. swab/bedsore 2.0 R R S S S R R R S S S R R R R S 9 — 7
18. Swab/bedsore 2.0 R S S S S R S R S S S S S R R R 6 — 10
19. Swab/wound 1.5 S R S S R R S S S S R S S R R R 7 — 9
20. Swab/wound 1.5 R R S S I R S S S S R S S R R S 6 1 9
21. Swab/wound 2.0 S R S S I R S S S S R S S R R S 5 1 10
22. Swab/wound 2.0 R S S S S R S R S S S S S R R R 6 — 10
23. Swab/wound 1.5 R S S S S R S I S S S S S R R S 4 1 11
24. Swab/wound 1.5 R S S S S R R R S S R S S R R R 8 — 8
25. Swab/ulceration 2.5 S S S S R R S R S S R S S R R S 6 — 10
26. Swab/ulceration 2.5 R S S S S R R R S S R S S R R R 8 — 8
27. swab/ulceration 2.5 R S S S S R S R S S S S S R R R 6 — 10
28. Swab/ulceration 2.0 R S S S S R S I S S S S S R R S 4 1 11
29. Swab/ulceration 2.0 R R S S I R S S S S R S S R R S 6 1 9
30. Swab/ulceration 2.0 R R S S S R R R S S S R R R R S 9 — 7

MZ, mezlocillin; PIP, piperacillin; TZP, piperacillin/tazobactam; MEM, meropenem.

144 SIENKIEWICZ ET AL.

FIG. 1. Standard strains susceptibility to thyme essential oil. MIC, minimal inhibitory concentration.

five isolates for which the MIC was 0.75 ml/ml, isolated from
diverse materials, and with different susceptibility to tested
drugs.
The increasing prevalence of methicillin resistance among
S. aureus strains is an increasing problem, one that has re-
newed interest in treating S. aureus infections with CC.
However, widespread use of MLSB antibiotics has led to an
increase of resistance to them.15 Although Prabhu et al. re-
ports that 28.42% of S. aureus strains are resistant to TE, our
results showed much greater resistance to TE as well as
CC.41 Resistant strains were a significant problem in our
study - 66.6% of the strains were resistant at the same time to
at least three antibiotics. However, they were sensitive to
glycopeptides and generally to LZD and chloramfenicol.
Only 10% of the strains that were susceptible/had interme-
diate susceptibility to all drugs.
Most of the resistant isolates occurred in the ICU, and the
major sites of isolation were swabs (wounds, nose, or ul-
ceration), which is in accord with the literature.41,57 For-
tunately, despite the fact that there were many multidrug
resistant strains, all of them showed susceptibility to ‘‘last
line’’ antibiotics.
In accordance with the literature, T. vulgaris L. oil was
shown to inhibit the growth of S. aureus strains isolated from
respiratory infections. The tested strains of S. aureus, sensi-
tive to 0.0125 ml/ml of oil, were resistant to oxacylin, GM,
and NN, and many of them, to NOR.19 Oil obtained from
Thymus fontanesii Boiss. Et Reut. containing carvacrol at a

FIG. 2. Clinical strains of Staphylococcus aureus, Enterococcus sp., Escherichia coli, and Pseudomonas aeruginosa susceptibility to
thyme essential oil.
THE ANTIMICROBIAL ACTIVITY OF THYME ESSENTIAL OIL
concentration of 0.3 ml/ml inhibited the growth of the stan-
dard and clinical strains of S. aureus isolated from clinical
materials.4 The oil was derived from the carvacrol chemo-
type of Thymus ciliatus (Desf.) Benth. ssp. eu-ciliatus Maire.
Thyme oil was also found to inhibit the growth of pristina-
micin-sensitive S. aureus strains isolated from respiratory
diseases with MIC 0.8 ml/ml; our results were similar.7 Re-
search on the antimicrobial properties of thyme oils obtained
from different chemotypes of Thymus spinulosus Ten. con-
firms the relationship between inhibitory activity and the
thymol and carvacrol content. These oils contained myrecene,
limonene, and g-terpinene as the dominant components and
low concentration of phenolic compounds. Their antimicrobial
activity was less effective, with MIC values ranging from 2.25
to 9.0 ml/ml against the standard strain of S. aureus ATCC
25923 and also against En. faecalis ATCC 29212.14
Many clinical strains of Enterococcus spp. were sensitive to
thyme oil at concentrations of 0.5 (n = 11) and 0.75 ml/ml
(n = 16), but En. faecalis isolate was inhibited at 1.25 ml/ml.
These strains came from diverse materials and hospital
wards. The En. faecium strains were, in general, more resis-
tant to antibiotics than En. faecalis strains. Fortunately, no
isolates were resistant to last-line drugs: glycopeptides, SYN,
and LZD. However, resistance to TE (n = 25, 83.3%), E (n = 19,
63.3%), CIP (n = 20, 66.6%), GM, and S (n = 17, 56.6%) was
common. Strains inhibited by 1.25 or 0.75 ml/ml of the es-
sential oil were isolated from different hospital wards and
presented diverse susceptibility to antibiotics.
In the last two decades, the importance of En. faecium as a
nosocomial pathogen has increased throughout the world
due to the greater ability of this species to acquire resistance
to drugs than En. faecalis. Very often, resistance among en-
terococci results from the presence of a putative pathoge-
nicity island. What is interesting is that an increase in
AM-resistant En. faecium usually precedes increasing rates of
VA-resistant strains.10,54 In our study, there were no isolates
showing resistance to VA, but 10 of 11 En. faecium strains
were resistant to AM. Regarding TE, although the entero-
cocci resistance rate decreased over time, it remains high in
some centers (57%).10 In our study, the resistance to TE was
even higher (83.3%), especially among En. faecalis strains. In
addition, erythromycin may be ineffective in therapy. Re-
sistance worldwide is very high, which is in accord with our
results.6,46 Overall, rates of high-level resistance to ami-
noglycosides were higher than those observed recently in
Brazil and in the United States.23,45
Due to the high content of active phenols and p-cymene,
the oil of T. vulgaris L. showed a very strong activity against
standard strains of Enterococcus genus, with MIC values
ranging from 0.0625 to 0.85 ml/ml in our studies. MIC values
obtained for the clinical strains cultured in the presence of
the oil were also significantly lower (0.25–1.25 ml/ml). Men-
tion is made within the literature of the inhibiting proper-
ties of thymol from Thymus sp. species on the adhesion of
S. aureus and Es. coli clinical strains to epithelial cells of
genitourinary system, which may be an alternative to syn-
thetic drugs in the prevention of urinary tract infections.12
We have shown that clinical strains of Es. coli were sensi-
tive to thyme oil at concentrations of 0.25 ml/ml (13 strains)
and 0.5 ml/ml (17 strains). They were more sensitive to oils
than P. aeruginosa, S. aureus, and Enterococcus spp. strains. As
far as susceptibility to antibiotics was concerned, many iso-
145
lates were resistant to AMP and TE (n = 21, 70%), CF (n = 19,
63.3%), SXT (n = 15, 50%), and CZ (n = 12, 40%). However, all
the isolates were sensitive to IPM, and most of them were
susceptible to ATM, CAZ, and NET. Strains inhibited by
0.5 ml/ml of essential oil were isolated from diverse materi-
als, in different hospital wards, and, generally, were more
resistant to antibiotics than strains inhibited by 0.25 ml/ml.
There was an increase observed in antimicrobial resistance
in Es. coli between 2002 and 2009. It was suggested that the
observed trends regarding resistance to third-generation
cephalosporins, a more than fivefold overall growth in re-
sistance (from 0.6% to 3.4%), may be the result of the addi-
tion of resistance traits to strains that were already
resistant.22 It may be explained by the spread of multidrug-
resistant plasmids that also contain genes for the production
of extended-spectrum beta-lactamase (ESBL).2,35 Resistant
strains were a significant problem in our study. The most
resistant strains came from wounds (n = 13) and were resis-
tant at the same time to even 11 or 14 of the tested drugs.
Contrary, there were two isolates (n = 6.6%) entirely suscep-
tible to antibiotics and seven that were resistant to one or two
of them (n = 23.3%).
Studies on the antimicrobial properties of the essential oil
obtained from T. fontanesii Boiss. Et Reut. containing carva-
crol explain its very strong activity against clinical strains of
Es. coli, with an MIC value of - 0.35 ml/ml.4 Similar MIC
values were obtained for the carvacrol chemotype of T. ci-
liatus (Desf.) Benth. ssp. eu-ciliatus Maire eu. against clinical
strains of colon bacilli isolated from the respiratory system.7
In our tests, clinical strains of Es. coli were sensitive to thyme
oil at concentrations of 0.25 and 0.5 ml/ml.
P. aeruginosa strains were sensitive to thyme oil at con-
centrations of 1.0–2.5 ml/ml. Many strains required 2.0–
2.5 ml/ml of the essential oil to be inhibited; therefore, they
were the most resistant when compared with other tested
bacterial species. At the same time, some isolates obtained
from bronchial secretions were resistant to all tested antibi-
otics (n = 2). Besides, the great majority of strains were re-
sistant to C (n = 28, 93.3%), STX (n = 27, 90%), AMC (n = 25,
83.3%), PIP and MZ (n = 21, 70%), and CTX (n = 20, 66.6%).
Fortunately, most isolates were susceptible to CAZ (n = 4),
GM (n = 5), IPM, and NET (n = 5). Eight strains with an MIC
of 2.5 ml/ml were isolated from diverse materials, and their
resistance to antibiotics was similar to isolates more sus-
ceptible to the essential oil.
A lot of published studies on antibiotic use and resistance
have reported increasing resistance among P. aeruginosa
strains,21 some show a decrease in CIP and CAZ resistance
among P. aeruginosa isolates as a result of decreased use of
those antibiotics.37 There is also a global problem with
MBLs P. aeruginosa strains, which are very often resistant to
other important groups of antibiotics tested, including third-
generation cephalosporins, aminoglycosides, and quino-
lones.8,13 In our study, there were some isolates resistant not
only to IPM but also to all the other antibiotics tested, which
could cause problems with therapy. In this situation, the only
therapeutic option may be polymyxins, which should not be
used as monotherapy.53
The most resistant to the tested oil were clinical strains of
P. aeruginosa, where inhibition of growth fell in the range of
1.5–2.5 ml/ml. Similar MIC values were obtained using the
disk-diffusion method for oil obtained from Thymus persicus
146
L. (thymol - 10%, carvacrol - 25%) and Thymus eriocalyx
(Ronniger) Jalas (thymol - 66%).24 The action of T. spinulosus
Ten. essential oil, which has a much lower content of active
phenolic compounds (thymol) than that of the oil derived
from T. vulgaris L., was much weaker against blue pus bacilli.
The obtained MIC values were within the limits of 4.5–
9.0 ml/ml, which was in accord with the literature.44 Other
species of thyme such as Thymus zygis L., T. serpyllum L.,
T. kotschyanus Boiss. & HoH., T. persisus L., and T. longicaulis
C. Presl also demonstrate antibacterial activity due to the
active phenol content.7,14
The literature reports not only the antimicrobial properties
of thyme and oregano essential oils, but also their antioxi-
dant properties as demonstrated by chemotypes from Thy-
mus munybyanus De Noe, T. pallescens De Noe, T. numidicus
Poiret, T. guyonii De Noe and oregano: Origanum glandulo-
sum Desf., and O. floribundum Munby.29 Studies on the bio-
logical properties of savory oil (Satureja hortensis L.),
containing phenols as predominant compounds, showed
antioxidant and antimicrobial properties against standard
strains of Gram-positive and Gram-negative bacteria.27
Due to the bactericidal and fungicidal properties of es-
sential oils, their use in pharmaceuticals and food are more
and more widespread as alternatives to synthetic chemical
products. Essential oils or some of their components are used
in perfume products, in sanitary products, in dentistry, in
agriculture, as food preservers and additives, and as natural
remedies.28,39 Essential oils are effective in anticancer ther-
apy, cardiovascular, and nervous system disorders; they
lower cholesterol levels, decrease and regulate glucose level,
and have antioxidative properties.17 Now, investigations on
the mechanism of action of essential oils and their compo-
nents are carried out both in vitro and in vivo on ani-
mals.5,38,47,48 Analytical monographs have been published
(National Pharmacopeia, European Pharmacopoeia, ISO,
WHO, Council of Europe) to ensure the necessary informa-
tion about essential oils: their source, concentration of active
components, and therapeutic doses. The cytotoxic capacity of
essential oils based on their pro-oxidant activity can make
them excellent antiseptic and antimicrobial agents. A big
advantage of essential oils is the fact that they are usually
devoid of long-term genotoxic risks. Moreover, some of them
show a very clear antimutagenic activity that could well be
linked to an anticarcinogenic activity. Recent studies have
demonstrated that the pro-oxidant activity of essential oils or
some of their constituents, as also that of some polyphenols,
is very efficient in reducing local tumor volume or tumor cell
proliferation by apoptotic and necrotic effects.32,36,50,51,56
However, essentials oils may be toxic not only against
bacteria, fungi, or viruses but also can have an adverse effect
on the human body when overdosed. In eukaryotic cells,
essential oils can provoke depolarization of the mitochon-
drial membranes by decreasing the membrane potential, af-
fect ionic Ca2 + cycling and other ionic channels, and reduce
the pH gradient.52 Some essential oils contain photoactive
molecules such as furocoumarins. The essential oils posses-
sing phototoxic activity are obtained from some plant fami-
lies, such as Apiaceae Rutaceae, Polygonaceae, and
Hypericaeae. This may cause damage to cellular macromol-
ecules and, in some cases, the formation of covalent adducts
to DNA, proteins, and cellular membrane.16 Some essential
oils or rather some of their constituents may be considered
SIENKIEWICZ ET AL.
secondary carcinogens after metabolic activation.26 For ex-
ample, psoralen, a photosensitizing molecule found in some
essential oils, for instance from Citrus bergamia, can induce
skin cancer after formation of covalent DNA adducts under
ultraviolet A or solar light.3 In addition, pulegone, a com-
ponent of essential oils from many mint species, can induce
carcinogenesis through metabolism generating the glutathi-
one depletory p-cresol.58
The application of essential oils in the treatment of many
human diseases, particularly infectious diseases caused by
multidrug resistant bacterial strains, may be an interesting
alternative for synthetic drugs that also show side effects.
Essential oils used in combination with antibiotics might
prevent antibiotic-resistant strain formation. Due to the
therapeutic problems associated with particularly resistant
strains, essentials oils can be useful in fighting diseases
caused by nosocomial pathogens.
Conclusions
Thyme oil obtained from T. vulgaris L.:
1. Shows very strong activity against standard and clinical
strains belonging to: Staphylococcus sp., Enterococcus sp.,
Escherichia sp., and Pseudomonas sp. genus.
2. It shows lower efficacy against clinical strains of P.
aeruginosa.
3. It is active against clinical strains resistant to most tes-
ted antibiotics.
Acknowledgments
The authors wish to thank Danuta Kalemba for thyme oil
analysis. The research reported in this article was supported
by grant no 503/5015-02/503-01 and has not been submitted
elsewhere.
Disclosure Statement
No competing financial interests exist.
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Address correspondence to:
Monika Sienkiewicz, Ph.D.
Medical and Sanitary Microbiology Department
Medical University of Lodz
Plac Hallera 1
90-647 Lodz
Poland
E-mail: monika.sienkiewicz@umed.lodz.pl