Antimicrobial, Multidrug Resistance Reversal and Biofilm Formation Inhibitory Effect of Origanum majorana Extracts, Essential Oil and Monoterpenes
Abstract
:1. Introduction
2. Results
2.1. Determination of the Composition of Essential Oil and n-Hexane Extract
2.2. Antibacterial Assay
2.3. Real-Time Ethidium Bromide Accumulation Assay
2.4. Biofilm Formation Inhibitory Effect
3. Discussion
4. Materials and Methods
4.1. Plant Material
4.2. Solvents and Chemicals
4.3. Preparation of the Extracts for the Bioassays
4.4. Essential Oil Distillation
4.5. Determination of the Composition of Essential Oil by GC and GC-MS
4.6. Bacterial and Fungal Strains and Culture Conditions for Antimicrobial Assay
4.7. Determination of Antibacterial Activity by Disc Diffusion Method
4.8. Determination of MIC Values
4.9. Bacterial Strains for Efflux Pump Inhibitory Assay
4.10. Real-Time Ethidium Bromide Accumulation Assay
4.11. Inhibition of Biofilm Formation
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
References
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Compounds a | RI b | % in Sample | |
---|---|---|---|
1 | α-thujene | 924 | 0.51 |
2 | α-pinene | 932 | 0.43 |
3 | sabinene | 972 | 4.53 |
4 | β-pinene | 978 | 0.26 |
5 | myrcene | 987 | 1.23 |
6 | α-phelladrene | 1007 | 0.14 |
7 | α-terpinene | 1017 | 3.00 |
8 | p-cymene | 1024 | 4.72 |
9 | limonene | 1028 | 0.20 |
10 | β-phelladrene | 1030 | 2.14 |
11 | β-ocimene | 1044 | 0.10 |
12 | γ-terpinene | 1057 | 6.48 |
13 | cis-sabinene hydrate | 1070 | 5.44 |
14 | terpinolene | 1085 | 1.62 |
15 | linalool | 1101 | 0.10 |
16 | trans-sabinene hydrate | 1103 | 25.18 |
17 | cis-p-menth-2-en-1-ol | 1125 | 2.35 |
18 | trans-p-menth-2-en-1-ol | 1142 | 1.34 |
19 | terpinen-4-ol | 1182 | 24.92 |
20 | α-terpineol | 1196 | 4.43 |
21 | linalyl acetate | 1249 | 1.92 |
22 | bornyl acetate | 1284 | 0.27 |
23 | terpin-1-en-4-yl acetate | 1295 | 0.36 |
24 | carvacrol | 1298 | 0.27 |
25 | trans-geranyl acetate | 1377 | 0.28 |
26 | β-caryophyllene | 1417 | 1.98 |
27 | spatulenol | 1574 | 0.47 |
Total | 94.67 |
Sample | S. aureus ATCC 29213 | S. aureus MRSA ATCC 43300 | S. epidermidis ATCC 12228 | E. faecalis ATCC 29212 | E. coli ATCC 35218 | E. coli AG-100 | K. pneumoniae ATCC 700603 | P. aeruginosa ATCC 27853 | C. albicans ATCC 10231 |
---|---|---|---|---|---|---|---|---|---|
MeOH extract | 7 ± 0.3 | 7 ± 0.0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
n-hexane extract | 7 ± 0.5 | 7 ± 0.3 | 7 ± 0 | 0 | 8 ± 0.2 | 7 ± 0 | 8 ± 0 | 7 ± 0.2 | 7 ± 0 |
Essential oil | 0 | 0 | 0 | 0 | 7 ± 0.2 | 0 | 7 ± 0.1 | 0 | 0 |
Essential oil c | 16 ± 0.5 | 14 ± 0.3 | 14 ± 0 | 7 ± 0.3 | 14 ± 0.4 | 14 ± 0.4 | 10 ± 0.3 | 8 ± 0 | 13 ± 0.3 |
DMSO | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
Samples | S. aureus ATCC 25923 | S. aureus MRSA ATCC 43300 | E. coli ATCC 25922 | E. coli AG100 |
---|---|---|---|---|
MeOH extract | >100 µg/mL | >100 µg/mL | >100 µg/mL | >100 µg/mL |
n-hexane extract | >100 µg/mL | >100 µg/mL | >100 µg/mL | >100 µg/mL |
Essential oil | 0.125% | 0.125% | 0.125% | 0.250% |
Compounds | S. aureus ATCC 29213 | S. aureus MRSA ATCC 43300 | E. coli ATCC 35218 | E. coli AG100 |
---|---|---|---|---|
linalool | >10 µL/mL >56 mM | >10 µL/mL >56 mM | 10 µL/mL 56 mM | 10 µL/mL 56 mM |
sabinene | 10 µL/mL >62 mM | 10 µL/mL >62 mM | >10 µL/mL >62 mM | >10 µL/mL >62 mM |
sabinene-hydrate * | >0.154 mg/mL >62 mM | >0.154 mg/mL >62 mM | >0.154 mg/mL >62 mM | >0.154 mg/mL >62 mM |
α-terpinene | 10 µL/mL 61 mM | 10 µL/mL 61 mM | 10 µL/mL 61 mM | 10 µL/mL 61 mM |
γ-terpinene | >10 µL/mL >62 mM | >10 µL/mL >62 mM | >10 µL/mL >62 mM | >10 µL/mL >62 mM |
terpinen-4-ol | 10 µL/mL 60 mM | 10 µL/mL 60 mM | 5 µL/mL 30 mM | 5 µL/mL 30 mM |
Relative Fluorescence Index (RFI) 1 | ||||
---|---|---|---|---|
E. coli ATCC 25922 | E. coli AG100 | S. aureus ATCC 25923 | S. aureus MRSA ATCC 43300 | |
0.0625% | 0.125% | 0.0625% | 0.0625% | |
MeOH extract | −0.14 | −0.17 | −0.17 | −0.52 |
n-hexane extract | 0.04 | −0.12 | −0.06 | 0.39 |
Essential oil | 4.27 | 1.27 | 0.19 | 0.35 |
0.0312% | 0.0625% | |||
MeOH extract | −0.10 | −0.18 | ||
n-hexane extract | −0.09 | −0.13 | ||
Essential oil | 3.21 | 0.64 | ||
100 µM | 100 µM | 100 µM | 100 µM | |
α-terpinene | 0.07 | −0.00 | 0.06 | 0.00 |
γ-terpinene | 0.01 | −0.03 | 0.06 | −0.01 |
terpinen-4-ol | 0.07 | −0.00 | 0.02 | −0.01 |
sabinene | 0.25 | −0.02 | 0.06 | 0.10 |
sabinene hydrate | 0.02 | −0.04 | 0.00 | 0.27 |
linalool | −0.01 | −0.05 | 0.03 | −0.05 |
50 µM | 50 µM | |||
α-terpinene | 0.02 | 0.01 | ||
γ-terpinene | 0.05 | −0.01 | ||
terpinen-4-ol | 0.07 | −0.02 | ||
sabinene | 0.04 | −0.04 | ||
sabinene hydrate | −0.02 | −0.05 | ||
linalool | −0.02 | −0.08 | ||
CCCP 2 | 2.00 | 0.70 | ||
RES 3 | − | 1.55 | 1.06 |
Sample | Inhibition % | |||
---|---|---|---|---|
E. coli ATCC 25922 | E. coli AG100 | |||
0.0625% | 0.0312% | 0.125% | 0.0625% | |
MeOH extract | −5.77 ** | 55.61 * | −59.60 ns | −56.68 ns |
n-hexane extract | 57.79 * | 64.43 ns | −13.74 * | −41.58 ** |
Essential oil | −104.64 | – | 104.16 | – |
100 µM/ | 50 µM | 100 µM | 50 µM | |
α-terpinene | 1.95 * | 17.68 ** | −53.71 ns | −35.98 * |
γ-terpinene | 34.34 ** | 37.80 * | −33.37 ns | −13.62 ns |
terpinene 4-ol | −2.09 *** | 42.36ns | −28.19 ns | 8.51 ns |
sabinene | 36.35 ** | 48.57ns | −59.78 ns | 15.40 ns |
sabinene hydrate | 37.93 *** | 55.97 * | −42.47 ns | 12.13 * |
linalool | 28.98 *** | 49.68ns | −0.89 * | −11.78 * |
CCCP | 63.37 | 50.14 |
Sample | Inhibition % | |||
---|---|---|---|---|
S. aureus ATCC 25923 | S. aureus MRSA ATCC 43300 | |||
0.0625% | 0.0312% | 0.0625% | 0.0312% | |
MeOH extract | 0.93 ** | −1.93 ** | 44.99 ** | 39.69 * |
n-hexane extract | 1.22 ** | 0.46 ** | −53.03 ns | 18.37 ** |
Essential oil | 69.24 | – | 4.38 | – |
100 µM | 50 µM | 100 µM | 50 µM | |
α-terpinene | 1.47 ** | −1.13 ** | −64.81 ns | 26.82 ** |
γ-terpinene | 0.76 ** | −1.91 *** | −125.22 * | −53.03 * |
terpinene 4-ol | −0.64 ** | −1.51 ** | 66.31 ** | 53.71 ** |
sabinene | 0.61 ** | −1.83 *** | 53.53 ** | 86.26 * |
sabinene hydrate | −0.80 ** | −2.84 ** | 60.56 ** | 69.48 ** |
linalool | 0.21 ** | −2.58 ** | 34.44 ** | 28.87 * |
DMSO | 0.56 | −64.81 | ||
TZ | − | 97.07 | 94.66 |
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Ghazal, T.S.A.; Schelz, Z.; Vidács, L.; Szemerédi, N.; Veres, K.; Spengler, G.; Hohmann, J. Antimicrobial, Multidrug Resistance Reversal and Biofilm Formation Inhibitory Effect of Origanum majorana Extracts, Essential Oil and Monoterpenes. Plants 2022, 11, 1432. https://doi.org/10.3390/plants11111432
Ghazal TSA, Schelz Z, Vidács L, Szemerédi N, Veres K, Spengler G, Hohmann J. Antimicrobial, Multidrug Resistance Reversal and Biofilm Formation Inhibitory Effect of Origanum majorana Extracts, Essential Oil and Monoterpenes. Plants. 2022; 11(11):1432. https://doi.org/10.3390/plants11111432
Chicago/Turabian StyleGhazal, Tasneem Sultan Abu, Zsuzsanna Schelz, Lívia Vidács, Nikoletta Szemerédi, Katalin Veres, Gabriella Spengler, and Judit Hohmann. 2022. "Antimicrobial, Multidrug Resistance Reversal and Biofilm Formation Inhibitory Effect of Origanum majorana Extracts, Essential Oil and Monoterpenes" Plants 11, no. 11: 1432. https://doi.org/10.3390/plants11111432