EJBR2021v11i2art208 ISSN 2449-8955 European Journal of Biological Research Research Article European Journal of Biological Research 2021; 11(2): 203-211 DOI: http://dx.doi.org/10.5281/zenodo.4501542 Antimicrobial and antioxidant potentials, total phenolic contents of some herbal waters Sinem Aydin 1*, Ayşegül Caniklioğlu 2 1 Giresun University, Faculty of Arts and Sciences, Department of Biology, Giresun, Turkey 2 Giresun University, Faculty of Education, Department of Primary Education, Giresun, Turkey * Corresponding author: Phone: +90 454 310 40 41; Fax: +90 454 310 11 19; E-mail: sinem.aydin@giresun.edu.tr Received: 12 December 2020; Revised submission: 24 January 2021; Accepted: 04 February 2021 http://www.jbrodka.com/index.php/ejbr Copyright: © The Author(s) 2021. Licensee Joanna Bródka, Poland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/) ABSTRACT: The aim of the actual study is to evaluate antimicrobial and antioxidant potentials, total phenolic contents of thyme (Thymus sp.), myrtle (Myrtus communis L.), eucalyptus (Eucalyptus globulus L.) and rosemary herbal waters (Rosmarinus officinalis L.). They were bought a retailer in Giresun. In the studies, it was determined that only thyme water exhibited antimicrobial activity in all herbal waters. Streptomycine, tetracycline and nystatin which were synthetic antimicrobials demonstrated higher activity than studied herbal waters. Moreover; total flavonoid contents of the tested waters ranges from 50.19±0.0038 µ L CE/mL to 126.15±0.004 µ L CE/mL. The highest and the lowest total phenolic contents were detected in the thyme water and the eucalyptus water as 688.18±0.009 µ L GAE/mL and 24.54±0.0008 µ L GAE/mL, respectively. DPPH and ABTS radical scavenging activities of the herbal waters exhibited a dose dependent manner and increased with increasing conentrations. As a result of this study, it was concluded that thyme water could be an alternative to synthetic antimicrobial agents and thyme water, myrtle water, eucalyptus water and rosemary waters might be an alternative to synthetic antioxidative agents. Hence, further and detailed investigations are needed to determine active constituents in the herbal waters. Keywords: Herbal water; Antimicrobial activity; Bacteria; Fungus; Antioxidant activity; Oxidative stress. 1. INTRODUCTION Excessive utilization of antibiotic is harmful to human health and ecosystem. It might also leads to increase drug-resistant pathogens. Antibiotic resistance is a worldwide trouble which cause morbidity and mortality. It has been observed that many pathogenic bacteria gain resistance to the antimicrobial drugs quickly. Hence, multiple drug resistant bacteria caused the main failure in the treatment of infectious diseases. It is necessary to investigate and design the alternative drugs to combat resistant bacteria. Bioactive phytochemicals with antibacterial activity could be one of the alternative way to control multiple drug resistant bacteria. Owing to mechanism of action of plants differs from antibiotics, plants might neutralize resistant bacteria [1]. Oxidative stress leads serious diseases such as cancer, cardiovascular disease, neural disorders, Alzheimer's disease, mild cognitive impairment, Parkinson's disease, ulcerative colitis atherosclerosis and aging [2]. Antioxidants can be described as any substance that delays or hinders oxidative damage to a target Aydin & Caniklioğlu Antimicrobial and antioxidant activity of herbal waters 204 European Journal of Biological Research 2021; 11(2): 203-211 molecule. The main property of an antioxidant is its capability to trap free radicals. Antioxidant compounds such as phenols, polyphenols and flavonoids scavenge free radicals such as peroxide, or lipid peroxyl. Plants known as good antioxidant since ancient times [3]. The utilize of plants for the treatment of many illnesses on Earth is an ancient tradition that starts with humans preferring a settled life. Herbal remedies are significant part of the culture of rural communities in developing countries [4]. Herbal water describes the liquid acquired from the distillation of medicinal plants. It can also be known aqueous herbal extract. Different parts of plants, such as flowers, roots, seeds, leaves, fruits use to obtain herbal waters. Herbal waters are commonly utilized as dietary supplements and medicine [5]. The aim of the current study is to evaluate antimicrobial and antioxidant potencies, total phenolic contents of thyme (Thymus sp.), myrtle (Myrtus communis L.), eucalyptus (Eucalyptus globulus L.) and rosemary (Rosmarinus officinalis L.) herbal waters. 2. MATERIALS AND METHODS 2.1. Providing of the samples Myrtle water (commercial hydrosol), thymus water (commercial hydrosol), eucalyptus water (commercial hydrosol) and rosemary water (commercial hydrosol) were bought from a retailer in Giresun, Turkey. The scientific names of the herbs, common name and medicinal functions are detailes in Table 1. Table 1. Names and medicinal properties of herbal waters. Scientific name Use Thymus sp. Antimicrobial, appetite stimulant, astringent, anthelmintic and tonic. Thyme also utilizes against intestinal infections like hookworms, bacteria and fungi [6] Myrtus communis L. Antimicrobial, antioxidant, antimutagenic, astringent, antiseptic, anti-inflammatory, insecticide activities. Moreover, myrtle leaves are used in sweet liquors which have digestive features [7] Eucalyptus globulus L. It can be used as anesthetic, astringent, antiseptic, disinfectant, deodorant, expectorant, hemostat and vermifuge. It is also traditionally used in the treatment of various diseases like arthritis, asthma, bronchitis and wounds [8] Rosmarinus officinalis L. Rosemary diterpenes have also antioxidant property which inhibit neuronal cell death [9] 2.2. Microorganisms Eight bacteria and three yeast species were used in the study. Staphylococcus aureus (ATCC 29213) and Salmonella enterica (ATCC 14028) were obtained from Giresun Province Control Laboratory. Enterococcus faecalis (ATCC 29212) was obtained from were acquired from Rize University Department of Molecular Biology. Bacillus subtilis (ATCC 6633), Proteus vulgaris (ATCC 13315), Enterobacter aerogenes (CMM 2531), Candida albicans (FMC 17) and Candida tropicalis (ATCC 13803) were obtained from Fırat University Department of Biology. Gordonia rubripertincta (lab isolate) was obtained from Yeditepe University Department of Genetic and Bioenginneering. Klebsiella pneumoniae (ATCC 700603) and Candida parapsilosis (ATCC 22019) were obtained from Giresun University, Faculty of Education. 2.3. Determination of antimicrobial activities of herbal waters Disc diffusion method was used to reveal antimicrobial activity of herbal waters. Herbal waters were sterilized by using 0.45 μm pore sized filter. Standards antibiotics and antifungal agents (tetracycline, Aydin & Caniklioğlu Antimicrobial and antioxidant activity of herbal waters 205 European Journal of Biological Research 2021; 11(2): 203-211 gentamycine and nystatin) were used to compare inhibition zones. The turbidity of bacterial suspensions were adjusted 0.5 Mc Farland standard, then, the bacterial suspension inoculated into Müller Hinton Agar plates and allowed to dry. The turbidity of fungal suspensions were adjusted with 0.5 Mc Farland standard (107 CFU/mL fungi concentration), then the fungal suspensions spread petri dishes which contain Sabaroud Dextrose Agar and allowed to dry. The discs were put into agar plates. The discs (6 mm diameter) on the petri were impregnated with 20 μL of thymus water, myrtle water, eucalyptus water and rosemary water, separetely. The inoculated plates were standed in refrigerator for one hour. Then, plates were then incubated for 24 h at 37°C for bacteria and 48 h at 30°C for fungi. Diameter of zones were measured with a ruler. The sensitivity of the microorganisms to the studied waters was revealed by measuring the inhibitory zones size on the agar surface around the discs [10-13]. The tests were carried out three times. 2.4. Determination of antioxidant activities of herbal waters 2.4.1. Total phenolic content Total phenolic compounds of plant waters were determined with Folin–Ciocalteu reagent, according to the method of Slinkard and Singleton [14]. Aliquots (0.1 mL) of the herbal waters were transferred into test tubes and their volumes were made up to 4.6 mL with distilled water. After addition of 0.1 mL Folin– Ciocalteu reagent (previously diluted 3-fold with distilled water) and 0.3 mL 2% Na2CO3 solution, tubes were vortexed and the absorbance of the mixture was recorded after 2 h at 760 nm using a spectrophotometer. The quantity of the total phenolic content was denoted as µ L of gallic acid equivalent (GAE)/mL. The tests were carried out three times. 2.4.2. Total flavonoid content Total flavonoids of herbal waters were determined by the procedure of Zhishen et al. [15]. 0.25 mL herbal water was added to 1.25 mL distilled water followed by 75 µ L NaNO2 (5%) and incubated for 5 min. Afterwards, 150 µ L AlCl3.6H2O (10%) was incorporated to the mixture and further incubated for 5 min, the reaction mixture was treated with 0.5 mL NaOH (1 M) and 275 µ L distilled water. It was measured spectrometrically at 510 nm. The quantity of the total flavonoid content was denoted as µ L of cateschin equivalent (CE)/mL. The tests were carried out three times. 2.4.3. Total antioxidant capacity The total antioxidant capacity of the herbal waters was evaluated by the phosphomolybdenum method according to the procedure described by Prieto et al. [16]. 0.3 mL herbal water was combined with 3 mL of reagent solution (0.6 M sulfuric acid, 28 mM sodium phosphate and 4 mM ammonium molybdate). The absorbance of the reaction mixture was measured at 695 nm using a spectrophotometer. The quantity of the total antioxidant capacity was denoted as µ L of ascorbic acid equivalent (AAE)/mL. The tests were carried out three times. 2.4.4. 1,1‐Diphenyl‐2‐picryl‐hydrazyl (DPPH) radical scavenging activity Herbal waters were prepared at 250-1000 µ L/mL concentrations. 0.1 mL of each dilution was added to 3.9 mL of a 6x10-5 M methanolic solution of DPPH followed by vortexing. The mixture was shaken vigorously and allowed to stand in the dark at room temperature for 30 min. The decrease in absorbance of the resulting solution was measured spectrophotometrically at 517 nm against methanol [17]. The tests were Aydin & Caniklioğlu Antimicrobial and antioxidant activity of herbal waters 206 European Journal of Biological Research 2021; 11(2): 203-211 carried out three times. BHT and Rutin were used as standards. The DPPH radical scavenging activity was calculated using the following equation: DPPH radical scavenging activity (%) = [(A0 – A1) / A0] x 100 A0 is the absorbance of the control A1 is the absorbance of the sample 2.4.5. 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)(ABTS)radical scavenging activity Herbal waters were prepared at 250-1000 µ L/mL concentrations. 150 µ L herbal water of each dilution was mixed with 2850 µ L of the ABTS.+ solution for 2 h in the dark. Then the absorbance was taken at 734 nm using the spectrophotometer [18]. The ABTS.+ scavenging activity was calculated using the following equation: ABTS radical scavenging activity (%) = [(A0 – A1) / A0] x 100 A0 is the absorbance of the control A1 is the absorbance of the sample. 3. RESULTS AND DISCUSSION 3.1. Antimicrobial activity Disc diffusion method was use to determine the antimicrobial activity of the tested herbal waters. The inhibition zones of studied waters, standard antifungal and antibiotics are presented in Table 2. It was found that the myrtle, eucalyptus and rosemary waters didn’t show any antimicrobial activity against tested microorganisms. However, only thyme water inhibited bacteria with inhibition zone diameters ranging from 8-13 mm. B. subtilis was resistant to thyme water. Thyme water also had the highest and the lowest antifungal activity against C. parapsilosis and C. tropicalis, respectively. Streptomycine, tetracycline and nystatin which used standard antibiotic and antifungal agents had higher activity than tested herbal waters. Table 2. Inhibition zones of tested herbal waters, streptomycine, tetracycline and nystatin. Inhibition Zone (mm) Microorganisms TH MH EH DH Str. Tet. Nys B. subtilis -- -- -- -- 20.66±1.15 9.33±0.57 NT S. aureus 13.33±0.57 -- -- -- 14.33±1.15 15.66±0.57 NT E. faecalis 10.33±0.57 -- -- -- -- 17.66±0.57 NT G. rubripertincta 11.33±0.57 -- -- -- 20.66±1.15 15±1.00 NT E. aerogenes 8.33±0.57 -- -- -- 20.66±0.57 12.33±0.57 NT S. enterica 10.66±0.57 -- -- -- 13.33±0.57 15.33±0.57 NT P. vulgaris 11.66±0.57 -- -- -- 21.66±0.57 9.33±1.15 NT K. pneumoniae 11.33±1.15 -- -- -- 18.66±0.57 7.33±0.57 NT C. albicans 9.33±0.57 -- -- -- NT NT 22.66±1.15 C. tropicalis 9.33±0.57 -- -- -- NT NT 23.33±0.57 C. parapsilosis 11.66±0.57 -- -- -- NT NT 22±1.00 TH: Thymus water, MH: Myrtle water, EH: Eucalyptus water, RH: Rosemary water, Str: Streptomycine, Tet: Tetracycline, Nys: Nystatin, NT: Not Tested; (--): No inhibition. Values are expressed as means of three replicates ± SD. Herbal waters are commonly utilized in aromatherapy and researchs about their antimicrobial activities in vitro is limited. The antimicrobial effects of thyme, rosemary and mrytle waters have also been determined by other researchers. For example, Sağdıç indicated that two thyme (Thymus vulgaris L. and T. serpyllum L.) waters possessed a bactericidal action against Escherichia coli, E. coli O157H:7, Staphylococcus aureus and Aydin & Caniklioğlu Antimicrobial and antioxidant activity of herbal waters 207 European Journal of Biological Research 2021; 11(2): 203-211 Yersinia enterocolitica. In accordance with this study, we also found that thyme water had activity against S. aureus [19]. Yavuzer and Boğa found that thyme hydrosol had activity against S. aureus, Salmonella paratyphi and Klebsiella pneumoniae, Vibrio vulnificus, Pseudomonas luteola. We also found antimicrobial activity against S. aureus and K. penumoniae [20]. Oral et al. demonstrated that thyme, rosemary and mrytle hydrosols was active against Aeromonas hydrophila, Pseudomonas aeruginosa and Pseudomonas fluorescens. It was revealed also only thyme hydrosol active against E. coli [21]. Sağdıç and Özcan revealed that rosemary hydrosol were inactive against microorganisms including E. coli. Similarly, we also found no antimicrobial activity of rosemary water [22]. Hay et al. investigated antimicrobial activity of Colombian thyme and rosemary hydrosols against against E. coli, P. aeruginosa, S. aureus, Candida albicans and Aspergillus niger. Rosemery hydrosol didn’t present any activity against the bacteria and fungi up to 500 µ L/mL. Thyme hydrosol showed a minimum microbicidal activity of 250 µ L/mL against P. aeruginosa, S. aureus, C. albicans and A. niger. We also found that thyme water had activity against C. albicans and S. aureus but rosemary water had no activity against C. albicans and S. aureus [23]. 3.2. Antioxidant activity Flavonoids are secondary plant metabolites that possess substantial antioxidant and chelating features [24]. Total flavonoid content and total antioxidant capacity of the herbal waters were summarized in Table 3. The maximum total flavonoid content was found in thyme water (126.15±0.004 (µ L CE/mL) and the minimum total flavonoid content was found in rosemary water (50.19±0.0038 µ L CE/mL). Total antioxidant capacities of the herbal waters increase in the following order: Myrtle water < Thyme water < Rosemary water < Eucalyptus water. Table 3. Total flavonoid content (TFC) and total antioxidant capacity (TAC) of the herbal waters. Herbal waters TFC (µL CE/mL) TAC (µL AAE/mL) Thyme water 126.15±0.004 53.75±0.0016 Myrtle water 113.26±0.0004 15.5±0.0024 Eucalyptus water 82.88±0.0002 163.1±0.097 Rosemary water 50.19±0.0038 77.85±0.0075 Values are expressed as means of three replicates ± SD. DPPH solution exhibits demonstrates a deep purple colour at 517 nm. This purple colour usually discolours when antioxidant molecules scavenges DPPH radicals and turns into them a bleached product [24]. DPPH radical scavenging activities of the herbal waters are given in Table 4. DPPH radical scavenging activity of the tested waters and standards (Rutin and BHT) increased steadily with increasing concentration of samples. The highest DPPH radical scavenging activity were observed in myrtle water (54.50±0.012) and the lowest DPPH radical scavenging activity were observed eucalyptus water (16.53±0.015) at 1000 µ L/mL concentrations. All plant hydrosols have lower activity than standards (BHT and Rutin). ABTS radical scavenging activities of the tested herbal waters are illustrated in Table 5. ABTS radical scavenging activities of the waters were compared with the standards BHT and Rutin. The strongest antioxidant properties, determined by ABTS radical scavenging assay, were in thyme water even better than standard antioxidants. Activities of the herbal waters increase with the increasing concentration. Rutin and BHT exhibited higher activity than the tested waters except for thyme water. ABTS radical scavenging Aydin & Caniklioğlu Antimicrobial and antioxidant activity of herbal waters 208 European Journal of Biological Research 2021; 11(2): 203-211 activity of the herbal waters and standards increased in the following order: Thyme water > BHT > Rutin > Rosemary water > Eucalptus water > Myrtle water. Table 4. DPPH scavenging activity of the herbal waters and standards. Herbal Water Concentration (µL/mL) (% inhibition) DPPH scavenging activity Eucalyptus water 250 500 750 1000 2.68±0.005 8.12±0.014 14.08±0.010 16.53±0.015 Rosemary water 250 500 750 1000 12.64±0.008 14.80±0.013 22.03±0.007 22.72±0.004 Myrtle water 250 500 750 1000 40.42±0.012 42.80±0.017 46.35±0.017 54.50±0.012 Thyme water 250 500 750 1000 23.27±0.004 38.02±0.006 50.56±0.007 50.74±0.012 Rutin 250 500 750 1000 88.17±0.005 88.59±0.009 90.21±0.005 92.25±0.002 BHT 250 500 750 1000 84.62±0.007 86.39±0.004 89.49±0.009 90.50±0.007 Values are expressed as means of three replicates ± SD. Table 5. ABTS radical scavenging activity of the herbal waters and standards. Herbal water Concentration (µL/mL) (% inhibition) DPPH scavenging activity Eucalyptus water 250 500 750 1000 13.95±0.019 23.66±0.027 28.29±0.013 31.93±0.019 Rosemary water 250 500 750 1000 16.80±0.852 25.63±0.725 29.31±0.684 35.85±0.619 Myrtle water 250 500 750 1000 10.93±0.023 17.36±0.009 21.23±0.019 23.51±0.010 Thyme water 250 500 750 1000 96.66±0.0004 99.04±0.0006 99.54±0.0004 99.65±0.0002 Rutin 250 500 750 1000 78.54±0.048 81.94±0.019 85.26±0.010 87.63±0.006 BHT 250 500 750 1000 93.48±0.011 93.92±0.006 94.43±0.004 96.65±0.008 Values are expressed as means of three replicates ± SD. Aydin & Caniklioğlu Antimicrobial and antioxidant activity of herbal waters 209 European Journal of Biological Research 2021; 11(2): 203-211 Antioxidant activity of thyme, rosemary and eucalptus hydrosols was searched by other researchers. Hay et al. found that rosemary and thyme hydrosols had ABTS radical scavenging activity [23]. Gharb revealed that Eucalyptus camaldulonsis hydrosol had DPPH radical scavenging activity and ferric reducing antioxidant potential (FRAP) [25]. Jeon et al. worked out antioxidant activity of rosemary hydrosols produced in Jeju. It was concluded that rosemary hydrosol had DPPH and ABTS radicals scavenging activity but it hadn’t Fe++ ion chelating activity [26]. We also found DPPH and ABTS radicals scavenging activities in thyme water, eucalyptus water and rosemary water. 3.3. Total phenolic content Total phenolic content of herbal waters were presented in Table 6. Total phenolic contents in the examined herbal waters ranged from 24.54±0.0008 µL GAE/mL to 688.18±0.009 µ L GAE/mL. The maximum total phenolic content was detected in thyme water and the minimum total phenolic content was found eucalyptus water. Table 6. Total phenolic contents (TPC) of herbal waters. Herbal water TPC (µL GAE/mL) Thyme water 688.18±0.009 Myrtle water 40.45±0.0003 Eucalyptus water 24.54±0.0008 Rosemary water 105.90±0.0008 Values are expressed as means of three replicates ± SD. 4. CONCLUSION Synthetic antioxidants and antimicrobials cause side effects to the body and doubts about their reliability cause studies on natural products in recent years. Phenolic compounds which found in plants and plant products is focused studies to plant essential oils and hydrosols. This study demonstrated that thyme water can be an alternative to synthetic antimicrobial agents and thyme water, myrtle water, eucalyptus water, rosemary water can be an alternative to antioxidant agents. The presented results will be the base for future research but it is needed detailed investigations for a better understanding of the active compounds involved in herbal waters. Authors' Contributions: SA: studied antioxidant activity of the herbal waters and structured the paper. AC: studied antimicrobial activity of the herbal waters and edited the manuscript. Both authors read and approved the final manuscript. Conflict of Interest: The authors declare that they have no conflict of interest. REFERENCES 1. Masoumian M, Zandi M. Antimicrobial activity of some medicinal plant extracts against multidrug resistant bacteria. Zahedan J Res Med Sci. 2017; 19(11): e10080. 2. Pisoschi AM, Pop A, Cimpeanu C, Predoi G. Antioxidant capacity determination in plants and plant-derived products: A review. Oxid Med Cell Longev. 2016; 2016: ID 9130976. 3. Mahdi-Pour B, Jothy LS, Latha LY, Chen Y, Sasidharan S. Antioxidant activity of methanol extracts of different parts of Lantana camara. Asian Pac J Trop Biomed. 2012; 2(12): 960-965. Aydin & Caniklioğlu Antimicrobial and antioxidant activity of herbal waters 210 European Journal of Biological Research 2021; 11(2): 203-211 4. Njume C, Afolayan AJ, Ndip RN. An overview of antimicrobial resistance and the future of medicinal plants in the treatment of Helicobacter pylori infections. Afr J Pharm Pharmacol. 2009; 3: 685-699. 5. Tabaraki R, Sadeghinejad N. Comparison of the antioxidant activity of volatile compounds of traditional herbal waters per serving cup. Iran J Pharm Sci. 2013; 9(1): 47-54. 6. Reddy VP, Vital KR, Varsha PV, Satyam S. Review on Thymus vulgaris traditional uses and pharmacological properties. Med Aromat Plants. 2014; 3: 164-166. 7. Pirbalouti AG, Mirbagheri H, Hamedi B, Rahimi E. Antibacterial activity of the essential oils of myrtle leaves against Erysipelothrix rhusiopathiae. Asian Pac J Trop Biomed. 2014; 4(Suppl. 1): 505-509. 8. Bachir RG, Benali M. Antibacterial activity of the essential oils from the leaves of Eucalyptus globulus against Escherichia coli and Staphylococcus aureus. Asian Pac J Trop Biomed. 2012; 2(9): 739-742. 9. Habtemariam S. The therapeutic potential of rosemary (Rosmarinus officinalis) diterpenes for Alzheimer’s disease. Evid Based Complem Altern Med. 2016; 6: 1-14. 10. Murray PR, Baron EJ, Pfaller MA, Tenovar FC, Yolke RH, Washington DC, ASM Press, 1995. 11. Saric CL, Cabarkapa SI, Beljkas MB, Misan CA, Sakac BM, Plavsic VD. Antimicrobial activity of plant extracts from Serbia. Food Process Qual Safety. 2009; 1(2): 1-5. 12. Ertürk Ö. Antibacterial and antifungal activity of ethanolic extracts from eleven spice plants. Biologia Bratislava. 2006; 61(3): 275-278. 13. Ünal MÜ, Uçan F, Şener A, Dinçer S. Research on antifungal and inhibitory effects of DL-limonene on some yeasts. Turk J Agric For. 2008; 36: 576-582. 14. Slinkard K, Singleton VL. Total phenol analysis: automation and comparison with manual methods. Am J Enol Viticult. 1977; 28: 49-55. 15. Zhishen J, Mengcheng T, Jianming W. The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chem. 1999; 64: 555-559. 16. Prieto P, Pineda M, Aguilar M. Spectrophotometric quantitation of antioxidant capacity through the formation of a phosphor molybdenum complex: Specific application to the determination of vitamin E. Anal Biochem. 1999; 269: 337-341. 17. Blois MS. Antioxidant determinations by the use of a stable free radical. Nature. 1958; 26: 1199‐1200. 18. Arnao MB, Cano A, Acosta M. The hydrophilic and lipophilic contribution to total antioxidant activity. Food Chem. 2001; 73: 239‐244. 19. Sagdic O. Sensitivity of four pathogenic bacteria to Turkish thyme and oregano hydrosols. Lebensm Wiss Technol. 2003; 36: 467-473. 20. Yavuzer E, Boğa EK. Testing the antimicrobial effects of some hydrosols on food borne-pathogens and spoilage bacteria. J Limnol Freshw Fish Res. 2020; 6(1): 47-51. 21. Oral N, Vatansever L, Güven A, Gülmez. Antibacterial activity of some Turkish plant hydrosols. Kafkas Üniv Vet Fak Derg. 2008; 14(2): 205-209. 22. Sağdıç O, Özcan M. Antibacterial activity of Turkish spice hydrosols. Food Cont. 2003; 14: 141-143. 23. Stankovic MS. Total phenolic content, flavonoid concentration and antioxidant activity of Marribium peregrinum L. extracts. Kragujevac J Sci. 2011; 33: 63-72. 24. Hay YO, Sierra MAA, Tellez M, Sequeda LG, Tellez AN, Bonnafaus C, Raynaud C. Phytochemical, antioxidant and antimicrobial parameters of essential oils and hydrosols of Colombian thyme and rosemary obtained using two different steam distillation methods. Int J Phytocos Nat Ingred. 2015; 2(7): 1-10. Aydin & Caniklioğlu Antimicrobial and antioxidant activity of herbal waters 211 European Journal of Biological Research 2021; 11(2): 203-211 25. Gharb LA. Comparison between the antioxidant activity of volatile oil and hydrosol in Eucalyptus camaldulensis (young and adult) leaves. Indian J Public Health Res Dev. 2020; 11(1): 1124-1128. 26. Jeon DH, Moon JY, Hyun HB, Cho SK. Composition analysis and antioxidant activities of the essential oil and the hydrosol extracted from Rosmarinus officinalis L. and Lavandula angustifolia Mill. J Appl Biol Chem. 2013; 56(3): 141-146.