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Article

Antimicrobial and Antioxidant Activity of Different Honey Samples from Beekeepers and Commercial Producers

by
Miroslava Kačániová
1,2,*,
Petra Borotová
3,
Lucia Galovičová
1,
Simona Kunová
4,
Jana Štefániková
3,
Przemysław Łukasz Kowalczewski
5 and
Peter Šedík
6
1
Institute of Horticulture, Faculty of Horticulture and Landscape Engineering, Slovak University of Agriculture, Tr. A. Hlinku 2, 94976 Nitra, Slovakia
2
Department of Bioenergy, Food Technology and Microbiology, Institute of Food Technology and Nutrition, University of Rzeszow, Zelwerowicza St. 4, 35601 Rzeszow, Poland
3
AgroBioTech Research Centre, Slovak University of Agriculture, Tr. A. Hlinku 2, 94976 Nitra, Slovakia
4
Institute of Food Sciences, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture, Trieda A. Hlinku 2, 94976 Nitra, Slovakia
5
Department of Food Technology of Plant Origin, Poznań University of Life Sciences, 31 Wojska Polskiego St., 60-624 Poznań, Poland
6
Institute of Marketing, Trade and Social Sciences, Faculty of Economics and Management, Slovak University of Agriculture, Trieda A. Hlinku 2, 94976 Nitra, Slovakia
*
Author to whom correspondence should be addressed.
Antibiotics 2022, 11(9), 1163; https://doi.org/10.3390/antibiotics11091163
Submission received: 29 July 2022 / Revised: 19 August 2022 / Accepted: 27 August 2022 / Published: 29 August 2022
(This article belongs to the Special Issue Antioxidant and Antibacterial Properties of Honey)
Versions Notes

Abstract

:
Honey contains compounds with antioxidant and antibacterial capacities, such as phenolic compounds and carotenoids. The current analysis evaluates the antioxidant and antibacterial activity of 100 honey samples from beekeepers from Slovakia and commercially purchased ones. Honey samples were diluted to 50%, 25%, 12.5%, and 6.25% concentrations. The antimicrobial activity of honey samples was evaluated against three Gram-positive, three Gram-negative bacteria, and four Candida spp. by well diffusion method. The highest antimicrobial effect of all honey concentrations was expressed as the size of the inhibition zone and was found against Pseudomonas aeruginosa among Gram-negative bacteria, Bacillus subtilis among Gram-positive bacteria, and Candida tropicalis among yeasts. Antibiotics used in the study showed the highest antimicrobial effect compared to all concentrations of honey samples. Slovakian honey from beekeepers and commercial honey samples from the Slovak market showed variable inhibitory effectiveness against microorganisms. The honey concentration of 50% was found the most effective. Lower concentrations of honey exhibited no effect against yeasts. The best antioxidant activity was found in a sample of buckwheat honey yielding 70.83% of DPPH inhibition and 2373.85 μg/g TEAC. Overall, better antioxidant activity was evaluated in honeydew honey.

1. Introduction

Honey is one of the major dietary components for humans, due to its therapeutic [1], antioxidant [2,3], antimicrobial [4,5], antitumoral [6], anti-inflammatory [7], antiviral [8], and antiulcer [5] activities.
The composition of honey, also in terms of the content of phenolic compounds, depends primarily on the botanical origin [9,10]. External factors, such as environmental conditions, harvesting season, storage, and processing method, also play an important role. Phenolic compounds are the most important antioxidants in honey. Besides having other effects, they are responsible for the therapeutic properties of honey which motivate its use in traditional and modern medicine for the treatment of human illnesses. Examples of the therapeutic applications of honey include facilitation of the treatment of diseases associated with oxidative stress, such as diabetes mellitus, hypertension, atherosclerosis, cancer, and Alzheimer’s disease [11]. In addition, due to the rich content of phenolic compounds in honey, the addition of honey may positively affect the organoleptic properties (color, taste, or flavor) of food [12]. Moreover, phenolic compounds were proposed as a chemical marker to determine the botanical and geographical origins of honey in a recent study [13].
Numerous studies focused on the antimicrobial potential of honey have been published to date [14,15,16]. The discovery of penicillin was one of the pivotal moments of modern medicine. However, with the rise of antibiotic-resistance in microbes, effectiveness of conventional antibiotics became threatened. Novel antibiotic-resistant forms have evolved within several bacterial genera, including Staphylococcus, Enterococcus, and Mycobacterium. Overuse of antibiotics makes the situation much worse by driving the occurrence of multidrug-resistant microbes [17,18]. Furthermore, high expenses associated with medication development hinder the production of new antimicrobial drugs to combat the emerging threat of antibiotic-resistant bacterial forms.
The aim of this study was to evaluate the antioxidant and antibacterial effect of honey from beekeepers and commercial bee honey samples of different floral origins against six strains of bacteria and four strains of yeast.

2. Results

2.1. Antimicrobial Activity of 50% Honey Samples

A 50% concentration of honey was tested against Gram-negative bacteria and the range of inhibition zones was determined in Table 1, Table 2 and Table 3. The antimicrobial activity against S. enterica ranged between 0.00 and 10.67 mm. The biggest inhibition zones were created by honey samples no. 31 and 88. No 31. was a multifloral honey from a beekeeper collected from forest and no. 88 was a commercial honey from linden. Streptomycin had a stronger antimicrobial effect (20.67 mm) than honey samples against S. enterica (Table 1). In 8 samples, antimicrobial activity of the 50% honey concentration against S. enterica was not observed at all (no. 19, 20, 23–28).
The antimicrobial activity of honey samples against Y. enterocolitica ranged between 0.00 and 10.67 mm. The highest value of inhibition was found in three samples, no. 31, 32, and 62. No. 31 was a multifloral honey collected from a forest and obtained from a beekeeper. No. 32 was also honey from a forest obtained from a beekeeper, but it was produced from honeydew. Sample no. 62 was honey of a rapeseed type from a countryside apiary obtained by a beekeeper. Also in this case, streptomycin had a stronger antimicrobial effect against Y. enterocolitica (14.67 mm) (Table 2). Y. enterocolitica was resistant against 53 honey samples where no antimicrobial activity was observed (Samples 1, 2, 4–12, 21–24, 34–37, 39–43, 49–52, 54–55, 69–76, 85–99).
The inhibition zones of honey samples created against P. aeruginosa ranged between 0.00 and 17.67 mm. The best antimicrobial activity was found in linden honey obtained from a beekeeper in the countryside. Antibiotic streptomycin resulted in a 20.33 mm inhibition zone against P. aeruginosa (Table 3). Sixty-eight honey samples did not show inhibitory effects against this microorganism (Samples no. 1–4, 13–31, 33–40, 47–49, 51–60, 67–68, 74–88, 90–96).
The antibacterial activity of the 50% honey was compared against the Gram-negative bacteria mentioned. Against S. enterica, the activity was found in 92% of honey samples, against Y. enterocolitica in 57%, and against P. aeruginosa in 32% which suggests that the most vulnerable G- bacteria was S. enterica and the most resistant was P. aeruginosa.
The sizes of the inhibition zones resulting from honey samples in the 25% concentration observed for Gram-negative bacteria are shown in Table 1, Table 2 and Table 3. The antimicrobial activity against S. enterica was observed only in 8 cases and ranged between 1.67 and 7.00 mm. The best antimicrobial activity was found in honey sample no. 31 which was a multifloral honey from a beekeeper obtained from the forest (Table 1). Among these honey samples, six were purchased from beekeepers and two were purchased from commercial shops.
The antimicrobial activity of the 25% honey concentration against Y. enterocolitica was found in 12 cases and inhibition zones ranged from 5.67 to 8.67 mm. The best result was observed in two samples, no. 29 was a honeydew honey from a beekeeper from a forest, and no. 61 a multifloral honey from a beekeeper from a city (Table 2). All samples which were effective in the 25% concentration were obtained from beekeepers.
The antimicrobial activity of the 25% concentration of honey samples against P. aeruginosa was found in 12 samples with zones ranging between 6.67 and 10.67 mm. The best antimicrobial activity was found in the linden honey obtained from a beekeeper in the countryside (Table 3). All samples that showed antimicrobial activity in the 25% concentration were obtained from beekeepers.
In the case of the 12.5% honey samples against Gram-negative bacteria, an antibacterial effect was found only against P. aeruginosa (Table 3). It was observed in 12 samples, and ranged between 4.33 and 8.33 mm. The best antimicrobial activity was found in multifloral honey from a beekeeper from a town. No antibacterial activity for the 6.25% concentration of honey samples was detected against Gram-negative bacteria.
The antimicrobial activity of honey samples against Gram-positive bacteria is shown in Table 4, Table 5 and Table 6. The inhibition zones created against S. aureus ranged between 0.00 and 12.33 mm (Table 4). The best antimicrobial activity was found in honey sample no. 3. It was honey from a beekeeper of multifloral origin collected in a town. Against S. aureus, a stronger antimicrobial effect was observed for the antibiotic chloramphenicol (22.33 mm) compared to honey samples (Table 4). In 82 samples, antimicrobial activity of the 50% honey against S. aureus was not observed.
The antimicrobial activity of honey samples at the 50% concentration against E. faecalis showed inhibition zones which ranged between 0.00 and 12.33 mm (Table 5). This highest value was found in sample no. 16, in honey from a beekeeper of multifloral origin collected from a town. The results were similar compared to S. aureus, but a larger antimicrobial effect was determined with the antibiotic chloramphenicol against E. faecalis (24.67 mm). No antimicrobial activity against this microorganism was found in 16 samples.
The antimicrobial activity of honey samples against B. subtilis ranged between 0.00 and 12.67 mm (Table 6). The best antimicrobial activity was found in chestnut honey from a beekeeper collected from the forest. In the case of chloramphenicol, the diameter of the inhibition zone was 22.33 mm. In total, 23 honey samples did not show an effect against B. subtilis.
The antibacterial activity of the 50% honey against S. aureus was found only in 18% of honey samples, against E. faecalis in 84%, and against E. faecalis in 77%. This result implies the best activity of honey against E. faecalis. The ranges of the inhibition zones of 25% of honey samples in the tested Gram-positive bacteria are shown in Table 4, Table 5 and Table 6. Against S. aureus, 13 samples of honey showed antimicrobial activity which ranged between 2.38 and 13.00 mm. The best antimicrobial activity was found in honey sample no. 4, which was multifloral honey from a beekeeper from a town (Table 4). All antibacterial activities of the 25% honey against S. aureus were found in samples obtained from beekeepers.
The antimicrobial activity of honey samples against E. faecalis was found only in 4 samples of the 25% honey and ranged between 10.00 and 11.00 mm. The highest value was found only in sample no. 16, in multifloral honey from a beekeeper from a town (Table 5). All samples which were effective in the 25% concentration against E. faecalis were from beekeepers.
The antimicrobial activity of honey samples against B. subtilis was found in 9 samples and ranged between 4.33 and 14.33 mm. The best antimicrobial activity was found in sample no. 4, a multifloral honey from a town obtained from a beekeeper (Table 6). All samples which showed antimicrobial activity in the 25% concentration against B. subtilis were from beekeepers.
The antibacterial activity of the 12.5% honey was found against all three Gram-positive bacteria species. The antibacterial effect against S. aureus was observed in 9 samples with the best activity observed in sample no 4. It was a beekeeper’s honey of multifloral origin from a town (Table 4). Honey in the 6.25% concentration showed an antibacterial effect only in two samples. No. 4 had an inhibition zone of 5.33 mm (multifloral honey from a beekeeper from a town). Sample no. 53 had an inhibition zone of 3.33 mm (fruit-tree honey from a beekeeper from a countryside).
The antimicrobial effects of the 12.5% honey against E. faecalis were found in four samples and ranged between 5.67 and 8.33 mm. The best antimicrobial activity was observed in acacia honey samples from a beekeeper from a town (Table 5). Honey in the 6.25% concentration did not show any antimicrobial activity against E. faecalis.
Against B. subtilis, antimicrobial activity was found in 8 samples and ranged between 3.00 and 11.67 mm. The best antimicrobial effect was found in multifloral honey samples from a beekeeper from a town (Table 6). In the 6.25% concentration, only one sample showed an antagonistic effect against this microbe which was sample no. 4, multifloral honey from a beekeeper from a town.
Values of the inhibition zones for the 50% honey samples in the tests against yeasts are shown in Table 7, Table 8, Table 9 and Table 10. The antimicrobial activity against C. albicans ranged from 0.00 to 7.67 mm. The best antimicrobial activity was found in honey samples no. 1 and 65. No 1. was a multifloral honey from a beekeeper from a town and no. 65 was a sunflower honey from a beekeeper of the countryside. Comparing the activity of honey to antifungal fluconazole, a stronger antimicrobial effect was found in fluconazole (20.33 mm) (Table 7). In 18 samples of the 50% honey, antimicrobial activity against C. albicans was not found.
The antimicrobial activity of the honey samples against C. glabrata ranged from 0.00 to 10.00 mm (Table 8). The largest zone of inhibition was found in sample no. 100, phacelia honey from a beekeeper collected in the countryside. A larger antimicrobial effect was found against C. glabrata for fluconazole (24.67 mm). Antimicrobial activity against C. glabrata was not observed in 19 samples.
The antimicrobial activity of honey samples against C. krusei ranged between 0.00 and 9.67 mm. The best antimicrobial activity was found in fruit-tree honey from a beekeeper collected in the countryside. The susceptibility of C. krusei to fluconazole was higher (22.33 mm) (Table 9). Ten honey samples did not show any effects against C. krusei.
The inhibition zones of honey samples against C. tropicalis ranged from 0.00 to 10.67 mm. The inhibition zone of fluconazole against this yeast was 21.33 mm (Table 10). Susceptibility of C. tropicalis to the antifungal compound was stronger than to honey samples. Seven samples of honey were not effective against C. tropicalis.
The anticandidal effect of honey in the 50% concentration was found in 82% of honey samples against C. albicans, in 81% of honey samples against C. glabrata, in 90% of honey samples against C. krusei, and in 93% of honey against C. tropicalis. Honey samples in 50% concentration were more effective against Candida species compared to bacterial species.
The anticandidal effect of the 25% concentration of honey was found for each candida only in the sample no. 100 which was phacelia honey from a beekeeper from a countryside apiary. The sizes of inhibition zones were: 6.33 mm against C. tropicalis, 3.33 mm against C. glabrata, 1.00 mm against C. albicans, and 0.33 mm against C. krusei.

2.2. Antioxidant Activity of Honey

The level of DPPH inhibition ranged from 8.51% to 70.83% which corresponded to 277.71–2373.85 μg Tx/g TEAC. A stronger activity, higher than 1000 μg Tx/g TEAC, was observed for 13 samples (sample numbers 64, 95, 28, 32, 34, 33, 27, 35, 94, 69, 93, 90, and 81). A very strong activity was determined only in honey sample 64 with 70.83% inhibition which corresponded to 2373.85 μg Tx/g TEAC (Table 11).
With respect to the type of honey, the highest activity was determined in buckwheat honey samples with an average activity of 53.11% of DPPH and 1758.19 μg Tx/g TEAC. A very good activity was also found in the group of honeydew honey samples, where all the samples showed high activity in the range from 29.84 to 41.94% and TEAC ranging from 962.49 to 1354.72 μg Tx/g. One sample of manuka honey showed an activity value of 34.02% which corresponded to 1097.52 μg Tx/g TEAC. The lowest activity was determined in honey samples from acacia where the activity reached levels ranging from 8.51 to 16.96% and 277.71 to 550.98 μg Tx/g TEAC (Table 12).

3. Discussion

Antibiotic resistance is a pressing concern for modern healthcare. According to resistance surveillance studies, resistance to frequently used antibiotics is rising. Honey has been valued for its medicinal properties since ancient times [19]. The present study was performed in order to evaluate the antimicrobial potential of honey produced in Slovakia by beekeepers and honey purchased commercially in Slovak markets against six pathogenic bacterial and four yeast species. The strongest antibacterial activity against Gram-negative bacteria of the 50% concentration honey samples was found towards P. aeruginosa (17.67 mm). Among the tested Gram-positive bacteria, honey in the 50% concentration was found the most active against B. subtilis (13.00 mm). In the case of yeast and the same honey concentration, C. tropicalis was the most susceptible (10.67 mm). Overall, the honey samples showed small differences in the results. In the study of Wadi [20], honey was evaluated against 8 clinical isolates including Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus vulgaris, Salmonella typhi, Shigella sonnei, and methicillin-resistant Staphylococcus aureus (MRSA). Raw natural and commercial honey from the study exhibited antibacterial properties against tested Gram-positive and Gram-negative bacteria.
Our results revealed that both honey from beekeepers and commercial honey showed identical activity towards most of the tested microorganisms and the same results were revealed in the study of Wadi [20]. Our data imply that the connection between antibacterial activity, floral source, and environmental parameters varies by geographical area. Similar results were found in the study by Wahdan [21]. Antibacterial activity of different floral types of honey was examined against a wide range of Gram-positive and Gram-negative bacterial standard organisms and significant antimicrobial activity was proved [22].
In a study that compared undiluted honey and honey diluted at concentrations of 75, 50, 30, and 10%, it was found effective against S. aureus and S. epidermidis [23]. In our study, where the antimicrobial activity of 100 honey samples diluted to 6.25, 12.5, 25, and 50% concentrations was compared to the effect of three antibiotics (streptomycin, chloramphenicol, and fluconazole), the results were similar. Honey samples with the 25% concentration showed very good antimicrobial potential against Gram-negative bacteria (P. aeruginosa). Results of tests performed with the well diffusion method showed the highest antimicrobial activity at 10.67 mm. The best antimicrobial activity of honey at a concentration of 25% against Gram-negative bacteria was found against B. subtilis with an inhibition zone of 14.33 mm. Comparative research of antibacterial activity of honey in various concentrations determined better activity compared to commonly used antibiotics against nine pathogens from urine samples [24]. An in vitro study on local Nigerian types of honey showed inhibition activity of tested undiluted and diluted samples against enteropathogenic isolates [25,26]. Three honey samples were obtained from Baghdad and were examined for antibacterial activity. Three concentrations of 100, 70, and 50% were used against different organisms. The highest antibacterial activity of honey was reported in a 100% concentration [27]. The activity of Danish honey was mostly considered due to the hydrogen peroxide content [28]. The difference in antibacterial activity of honey greatly depends on its floral origin [29]. Honey rich in multiflora improves antimicrobial properties against several clinically significant microorganisms. Moreover, it increases its nutritional potential [30].
The next used concentrations of honey were 12.5 and 6.25%. Among the tested Gram-negative bacteria, an inhibitory effect of 12.5% honey was found towards P. aeruginosa in 12 samples. The honey at a concentration of 6.25% inhibited the growth P. aeruginosa only in two samples. Honey at a lower concentration was more effective against Gram-positive bacteria. Similar to the results at higher concentrations, B. subtilis was inhibited to the greatest extent. Only a few samples of honey at 6.25% concentration showed an antibacterial effect. Compared to antibiotic gentamicin, honey was found more effective as an antibacterial agent against Pseudomonas and Staphylococcus strains [31]. A synergistic action of honey and antibiotics was demonstrated, and it was suggested that honey can be given orally with antibiotics [32]. Our study demonstrated very good activity in all concentrations against P. aeruginosa and B. subtilis. The effect of antibiotics on different microorganisms was higher than that of the honey samples.
The best antimicrobial effect was found for multifloral, linden, chestnut, and honeydew honey samples from beekeepers. Furthermore, some multifloral commercial honey samples showed very good activity. As it was mentioned previously, geographical and botanical sources had pronounced effects on the antibacterial properties of honey. The antibacterial activity of honey is influenced by a variety of factors besides the floral source. Honey could be used as an alternative treatment for chronic wounds and burns as it can inhibit different types of microorganisms which do not respond to conventional antibiotics without side effects [20]. In research by Goślinski et al. [33], honeydew honey showed a total content of polyphenols comparable to manuka honey, which was also much higher than that detected in multifloral and linden honey. In a different study, manuka honey, alongside honeydew honey, showed a stronger antimicrobial effect against Gram-positive than against Gram-negative bacteria. Similar results were found in our study. High antibiotic activity of manuka honey was found against the strains of S. aureus and E. faecalis in our results. Other researchers also reported that Gram-positive bacteria are more sensitive than Gram-negative microorganisms to the bactericidal activity of honey [4,34].
The antimicrobial property of honey was found to increase with an increasing concentration of honey [35]. This concentration dependency was confirmed in our study. Similar dose-dependent antibacterial activity of honey was also observed by Deng et al. [36] and Ghramh et al. [37] in studies on the antibacterial potential of honey obtained from different nectars. Based on different studies, it is possible to conclude from that variation in antibacterial potential of honey could depend on its botanical and geographical origin, storage conditions, and metabolism of honeybees [37,38,39,40,41].
Honey is rich in polyphenols, vitamins, and enzymes, and shows good antioxidant activity [42]. In our study, the highest antioxidant capacity was found in buckwheat honey with 70.83% inhibition which was equivalent to 2373.85 μg Tx/g honey. A strong activity of buckwheat honey was also determined in [43] where the authors compared 20 samples of buckwheat honey. In [44,45], the authors analyzed honey from different floral sources and found that buckwheat honey shows the best antioxidant properties. It is also known that buckwheat pollen and nectar are rich in antioxidants which is reflected in the antioxidant capacity of this honey [46]. The antioxidant activity of buckwheat honey was also studied in vivo where it was found to increase the antioxidant capacity of human serum [47].
Good antioxidant activities were also determined in honeydew types of honey where the highest activity reached 41.94% inhibition equivalent to 1354.72 μg Tx/g honey. Bobis et al. [48] found that the DPPH radical scavenging activity ranged from 47.84 to 62.99%. In another study [49], it was found that the antioxidant activity of honeydew honey was in the range from 40.67 to 64.83% DPPH inhibition which was the highest in comparison to acacia, lime, and sunflower types of honey. The authors also determined that the antioxidant activity positively correlated with the contents of total phenols and total flavonoids. The antioxidant capacity was tested in Spanish honey where the determined radical scavenging activity of honeydew honey was 66.8% on average; for comparison, 28.7% inhibition was reported for nectar honey [50]. The relationship between chemical composition and antioxidant activity was supported by other studies [51,52]. Moreover, in [53] it was stated that vitamins did not contribute to antioxidant capacity, while phenolic acids and flavonoids have an impact on the activity.
On the other hand, acacia samples of honey showed the lowest antioxidant activity in the range 8.51–16.96% and 277.71–550.98 TEAC. In [54], antioxidant activity of only 2.35–11.97% DPPH inhibition was reported. The authors also evaluated the activity of chestnut honey which was found to be in the range 0.95–3.54% inhibition. In our study, the chestnut honey samples showed an activity of 18.06–36.31% of DPPH radical inhibition. In the mentioned report, multifloral honey showed the strongest activity which ranged from 30.43 to 30.94% inhibition. In our study, the multifloral honey did not show better activity compared to the other tested honey samples with the highest activity of 25.22% inhibition.

4. Materials and Methods

4.1. Bee Honey Samples

One hundred various bee honey samples of different floral origins were obtained in 2020 from different apiaries as well from the local market where they were commercially sold under different brands. The beekeepers determined the floral source of the honey based on the availability of flora for nectar foraging, the location of the apiary, and the organoleptic qualities of the honey. Honey samples were stored in sterile glass jars at room temperature. Samples were labeled according to the source, location, and floral origin as shown in Table 13.

4.2. Microorganisms

Gram-negative bacteria (Pseudomonas aeruginosa CCM 3955, Yersinia enterocolitica CCM 7204, Salmonella enterica subsp. enterica 4420), Gram-positive bacteria (Bacillus subtilis CCM 1999, Staphylococcus aureus subsp. aureus CCM 2461, Enterococcus faecalis CCM 4224), and yeasts (Candida albicans CCM 8261, Candida glabrata CCM 8270, Candida krusei CCM 8271, Candida tropicalis CCM 8223) were obtained from the Czech Collection of Microorganisms (Brno, Czech Republic).

4.3. Determination of Antimicrobial Activity

The antimicrobial activity of each honey sample was determined using the well diffusion method. The inoculum was cultured for 24 h on Muller Hinton Broth (MHB, Oxoid, Basingstoke, UK) at 37 °C for bacteria and on Sabouraud Dextrose Broth (SDB, Oxoid, Basingstoke, UK) at 25 °C for yeast. In total, 100 µL of inoculum in a concentration of 0.5 McFarland standard (1.5 × 108 CFU/mL) was applied to a Petri dish (PD) with 20 mL of Mueller Hinton agar (MHA, Oxoid, Basingstoke, UK) for bacteria or Sabouraud Dextrose Agar (SDA, Oxoid, Basingstoke, UK) for yeasts. The following concentrations of the honey solutions were diluted with MHB resp. SDB: 6.25, 12.5, 25, and 50%. Subsequently, wells of 10 mm diameter were made with a sterile borer into agar plates containing the bacterial and yeast inoculum. In total, 20 μL of analyzed honey was added to the wells. The samples were incubated for 24 h at 37 °C for bacteria and 25 °C for yeast. Antibiotics (chloramphenicol, streptomycin, Oxoid, Basingstoke, UK) were used as a positive control for Gram-negative and Gram-positive bacteria. An antifungal (fluconazole, Oxoid, Basingstoke, UK) was used as a positive control for yeast. Disks impregnated with MHB served as a negative control. Inhibition zones were measured from the edge of the well to border of the bacterial growth at three sides. An inhibition zone above 10 mm was determined to be very strong antimicrobial activity, an inhibition zone above 5 mm was determined to be mild activity, and an inhibition zone above 1 mm was determined to be weak activity. Antimicrobial activity was measured three times.

4.4. Antioxidant Activity of Honey Samples

The antioxidant activity of honey samples was determined using the DPPH radical method. The DPPH (Sigma Aldrich, Schnelldorf, Germany) solution was prepared in methanol to a stock concentration 0.025 g/L and was adjusted using methanol to an absorbance of 0.8 at 515 nm (Glomax spectrophotometer, Promega Inc., Madison, WI, USA).
The 0.2 g of honey was mixed with 1 mL of distilled water. After that, 20 µL of suspension was added to 180 µL of DPPH solution in a 96-well plate. The samples were incubated on a shaker at 500 rpm for 30 min in the dark. The absorbance was measured at 515 nm. The % inhibition was calculated according to the formula:
%   o f   i n h i b i t i o n = A c o n t r o l − A   s a m p l e A c o n t r o l × 100
The control sample contained 20 µL of distilled water with 180 µL DPPH.
The total antioxidant capacity (TEAC) of honey samples was also calculated with the standard reference Trolox (Sigma Aldrich, Schnelldorf, Germany) prepared in methanol to 5 concentrations in the range of 20–100 µg/mL. TEAC was evaluated from the calibration curve as µg of Trolox to 1 g of honey sample.

5. Conclusions

Antimicrobial activity, especially with Gram-negative bacteria, Gram-positive bacteria, and yeasts, suggests that the honey under analysis may have a relevant role as natural antibacterial products that weaken the effects of bacterial infections and contribute to the improvement of food. In our study, four different concentrations of honey were studied. Our results showed that honey samples at 50% concentration had the strongest effect on the growth of yeast from the genus Candida. A lower concentration of honey (25%) produced in Slovakia had antibacterial activity against all Gram-negative and Gram-positive bacteria tested. Concentrations lower than 25% had an influence especially on P. aeruginosa, S. aureus, E. faecalis, and B. subtilis. The antioxidant activity was the highest in buckwheat honey. The majority of honey samples with good antioxidant properties were of the honeydew type.

Author Contributions

Conceptualization, M.K. and P.B.; methodology, M.K., P.B. and J.Š.; validation, M.K., P.B. and J.Š.; formal analysis, M.K., P.B., L.G., S.K. and J.Š.; resources, P.Š.; data curation, M.K., P.B., L.G., S.K., J.Š. and P.Š.; writing—original draft preparation, M.K., P.B., L.G., S.K., J.Š., P.Ł.K. and P.Š.; writing—review and editing, M.K. and P.Ł.K.; visualization, M.K.; supervision, M.K.; project administration, M.K. and P.Š. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by the Grant Agency of The Slovak University of Agriculture in Nitra, grant number 14-GASPU-2021 “Analysis of consumer behavior towards honeys enriched with health-promoting substances”.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Not applicable.

Acknowledgments

This publication was supported by the Operational program Integrated Infrastructure within the project: Sustainable smart farming systems taking into account the future challenges 313011W112, financed by the European Regional Development Fund.

Conflicts of Interest

The authors declare no conflict of interest.

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Table
Table 1. Antimicrobial activity of samples against S. enterica in mm.
Table 1. Antimicrobial activity of samples against S. enterica in mm.
Sample No.50%25%12.5%6.25%
16.33 ± 1.15---
26.67 ± 1.535.67 ± 0.58--
32.67 ± 0.58---
43.33 ± 0.58---
53.67 ± 0.58---
64.33 ± 0.58---
73.67 ± 0.58---
85.00 ± 1.00---
94.33 ± 0.58---
103.33 ± 0.58---
113.67 ± 0.58---
122.67 ± 0.58---
135.67 ± 0.58---
144.33 ± 0.58---
153.33 ± 0.58---
162.67 ± 0.58---
175.67 ± 0.58---
186.67 ± 0.58---
215.33 ± 1.15---
224.67 ± 0.58---
2910.33 ± 0.58---
309.67 ± 0.586.00 ± 1.00--
3110.67 ± 0.586.67 ± 0.58--
328.67 ± 0.587.00 ± 1.00--
335.33 ± 0.585.67 ± 0.58--
345.33 ± 0.58---
354.33 ± 0.58---
368.00 ± 1.00---
374.67 ± 0.58---
386.33 ± 0.58---
394.33 ± 0.58---
405.67 ± 0.58---
414.33 ± 0.58---
423.67 ± 0.58---
434.67 ± 0.58---
442.67 ± 0.58---
454.33 ± 0.58---
463.67 ± 0.58---
474.33 ± 0.58---
483.67 ± 0.58---
493.67 ± 0.58---
507.67 ± 0.58---
513.33 ± 0.58---
522.33 ± 0.58---
534.67 ± 0.58---
544.33 ± 0.58---
555.33 ± 0.58---
565.67 ± 0.58---
574.67 ± 0.58---
585.33 ± 0.58---
595.67 ± 0.58---
606.67 ± 0.58---
617.00 ± 1.00---
626.67 ± 0.58---
636.67 ± 0.58---
647.67 ± 0.58---
652.33 ± 0.58---
663.67 ± 0.58---
673.33 ± 0.58---
684.33 ± 0.58---
694.33 ± 0.58---
707.33 ± 0.58---
717.67 ± 0.58---
725.67 ± 0.58---
733.67 ± 0.58---
746.00 ± 1.00---
752.67 ± 0.58---
762.67 ± 0.58---
775.67 ± 0.58---
785.33 ± 0.58---
794.67 ± 0.58---
805.33 ± 0.58---
814.67 ± 0.58---
823.67 ± 0.58---
834.33 ± 0.58---
843.33 ± 0.58---
854.33 ± 0.58---
865.00 ± 1.00---
878.67 ± 0.584.67 ± 0.58--
8810.67 ± 0.584.33 ± 0.58--
893.33 ± 0.58---
903.67 ± 0.58---
913.33 ± 0.58---
922.67 ± 0.58---
933.67 ± 0.58---
944.67 ± 0.58---
954.67 ± 0.58---
963.33 ± 0.58---
976.67 ± 0.58---
985.33 ± 0.58---
993.67 ± 0.58---
1003.33 ± 0.581.67 ± 0.58--
ATB20.67 ± 1.15
Table
Table 2. Antimicrobial activity of samples against Y. enterocolitica in mm.
Table 2. Antimicrobial activity of samples against Y. enterocolitica in mm.
Sample No.50%25%12.5%6.25%
32.67 ± 0.58---
132.33 ± 0.58---
142.33 ± 0.58---
152.67 ± 0.58---
162.00 ± 1.00---
175.33 ± 0.58---
185.00 ± 1.00---
195.00 ± 1.00---
209.00 ± 1.006.67 ± 0.58--
255.33 ± 1.53---
264.67 ± 0.58---
273.67 ± 0.58---
283.67 ± 0.58---
2910.33 ± 0.588.67 ± 0.58--
309.67 ± 0.587.67 ± 0.58--
3110.67 ± 0.587.33 ± 0.58--
3210.67 ± 1.157.67 ± 0.58--
334.33 ± 0.58---
389.33 ± 1.158.33 ± 1.15--
442.33 ± 0.58---
452.33 ± 0.58---
462.67 ± 0.58---
472.33 ± 0.58---
482.33 ± 0.58---
534.33 ± 0.58---
563.67 ± 0.58---
574.33 ± 0.58---
584.67 ± 0.58---
592.67 ± 0.58---
603.67 ± 0.58---
6110.33 ± 0.588.67 ± 0.58--
6210.67 ± 0.588.33 ± 0.58--
638.67 ± 1.15---
647.33 ± 0.58---
655.67 ± 0.58---
6610.33 ± 0.587.67 ± 0.58--
6711.33 ± 0.586.00 ± 1.00--
689.33 ± 0.585.67 ± 1.15--
774.33 ± 0.58---
785.33 ± 0.58---
792.33 ± 0.58---
802.67 ± 0.58---
814.67 ± 0.58---
824.67 ± 1.15---
834.67 ± 0.58---
845.33 ± 1.15---
1006.33 ± 1.156.67 ± 1.53--
ATB14.67 ± 0.58
Table
Table 3. Antimicrobial activity of samples against P. aeruginosa in mm.
Table 3. Antimicrobial activity of samples against P. aeruginosa in mm.
Sample No.50%25%12.5%6.25%
57.00 ± 1.00---
68.67 ± 0.58---
75.00 ± 0.00---
86.33 ± 0.58---
913.33 ± 1.539.67 ± 0.587.67 ± 0.58-
1013.00 ± 1.0010.33 ± 0.588.33 ± 1.15-
1113.33 ± 1.539.33 ± 0.586.67 ± 1.15-
1210.67 ± 0.588.67 ± 0.585.00 ± 1.00-
325.33 ± 0.58---
415.33 ± 0.58---
427.67 ± 0.58---
436.67 ± 0.58---
445.67 ± 0.58---
4510.33 ± 0.58---
468.67 ± 0.5810.33 ± 0.586.33 ± 1.15-
505.67 ± 0.5810.33 ± 0.585.67 ± 0.58-
611.33 ± 0.58---
621.67 ± 0.58---
6311.67 ± 0.5810.33 ± 0.586.00 ± 1.00-
6410.33 ± 0.5810.33 ± 0.585.67 ± 0.58-
659.67 ± 0.5810.00 ± 1.006.00 ± 1.00-
666.67 ± 0.58---
6917.67 ± 2.5210.33 ± 0.584.33 ± 0.58-
7012.33 ± 0.5810.67 ± 0.585.33 ± 0.58-
715.67 ± 0.58---
7210.33 ± 0.58---
734.67 ± 0.58---
895.67 ± 0.58---
972.67 ± 0.58---
983.00 ± 1.00---
992.67 ± 0.58---
1004.67 ± 0.586.67 ± 1.536.67 ± 1.53-
ATB20.33 ± 0.58
Table
Table 4. Antimicrobial activity of samples against S. aureus in mm.
Table 4. Antimicrobial activity of samples against S. aureus in mm.
Sample No.50%25%12.5%6.25%
18.67 ± 0.584.67 ± 0.58
210.33 ± 0.585.00 ± 1.00
312.33 ± 0.588.67 ± 0.583.33 ± 0.58
410.33 ± 0.5813.00 ± 1.0010.33 ± 0.585.33 ± 0.58
188.33 ± 0.588.67 ± 0.58--
199.00 ± 1.009.33 ± 1.157.00 ± 1.00-
5311.00 ± 1.0011.00 ± 1.009.33 ± 0.583.33 ± 0.58
5411.00 ± 1.0011.00 ± 1.007.33 ± 0.58-
5511.67 ± 0.5810.33 ± 0.587.00 ± 1.00-
5610.67 ± 0.5810.67 ± 0.586.00 ± 1.00-
6911.00 ± 1.0011.33 ± 0.586.33 ± 1.15-
712.67 ± 0.58---
723.33 ± 0.58---
823.33 ± 0.58---
832.33 ± 0.58---
844.33 ± 0.58---
908.33 ± 0.5810.33 ± 0.585.33 ± 0.58-
917.67 ± 0.58---
1004.33 ± 0.582.33 ± 0.58--
ATB22.33 ± 0.58
Table
Table 5. Antimicrobial activity of samples against E. faecalis in mm.
Table 5. Antimicrobial activity of samples against E. faecalis in mm.
Sample No.50%25%12.5%6.25%
510.33 ± 0.58---
610.67 ± 1.15---
711.33 ± 0.58---
89.67 ± 0.58---
95.67 ± 0.58---
107.67 ± 0.58---
114.67 ± 0.58---
126.67 ± 0.58---
1311.67 ± 0.5810.33 ± 0.58--
1411.33 ± 0.5810.00 ± 1.73--
1511.00 ± 1.0010.33 ± 0.58--
1612.33 ± 0.5811.00 ± 1.00--
174.67 ± 0.58---
185.00 ± 1.00---
193.67 ± 0.58---
204.67 ± 0.58---
254.67 ± 1.15---
264.67 ± 0.58---
275.33 ± 0.58---
286.33 ± 0.58---
294.00 ± 1.00---
306.33 ± 0.58---
314.33 ± 0.58---
322.67 ± 0.58---
334.33 ± 0.58---
345.33 ± 0.58---
354.67 ± 0.58---
365.00 ± 1.00---
378.33 ± 0.58---
3810.33 ± 0.58---
3910.67 ± 0.58---
4011.33 ± 1.15---
455.67 ± 0.58---
466.67 ± 0.58---
475.33 ± 0.58---
488.33 ± 0.58---
493.67 ± 0.58---
504.33 ± 0.58---
512.67 ± 0.58---
523.67 ± 0.58---
537.67 ± 0.58---
546.67 ± 0.58---
556.33 ± 0.58---
565.67 ± 0.58---
577.67 ± 0.58---
588.67 ± 0.58---
597.67 ± 0.58---
607.33 ± 0.58---
655.67 ± 0.58---
664.67 ± 0.58---
673.67 ± 0.58---
684.33 ± 0.58---
694.67 ± 0.58---
705.33 ± 0.58---
712.33 ± 0.58---
722.67 ± 0.58---
735.67 ± 0.58---
746.00 ± 1.00---
755.33 ± 0.58---
766.33 ± 0.58---
775.67 ± 0.58---
784.67 ± 0.58---
793.33 ± 0.58---
803.67 ± 0.58---
814.33 ± 0.58---
824.33 ± 1.15---
835.00 ± 1.00---
844.67 ± 0.58---
8510.67 ± 0.58---
869.33 ± 0.58---
879.67 ± 0.58---
8810.33 ± 1.15---
895.00 ± 1.00---
904.67 ± 0.58---
915.33 ± 0.58---
924.67 ± 1.15---
934.33 ± 0.58---
944.67 ± 0.58---
953.67 ± 0.58---
963.33 ± 0.58---
975.67 ± 0.58---
986.33 ± 0.58---
995.33 ± 0.58---
1001.67 ± 0.58---
ATB24.67 ± 0.58
Table
Table 6. Antimicrobial activity of samples against B. subtilis in mm.
Table 6. Antimicrobial activity of samples against B. subtilis in mm.
Sample No.50%25%12.5%6.25%
111.00 ± 1.0011.00 ± 1.006.33 ± 0.58-
28.67 ± 0.5811.33 ± 1.158.33 ± 1.15-
35.33 ± 0.58---
413.00 ± 1.0014.33 ± 1.5311.67 ± 0.586.67 ± 0.58
52.67 ± 0.58---
95.33 ± 0.58---
102.33 ± 0.58---
115.33 ± 0.58---
126.00 ± 1.00---
156.67 ± 1.53---
166.33 ± 1.15---
194.67 ± 0.58---
215.33 ± 0.58---
222.33 ± 0.58---
239.67 ± 0.58---
246.67 ± 0.58---
254.67 ± 0.58---
275.33 ± 0.58---
282.33 ± 0.58---
306.00 ± 1.73---
316.33 ± 0.58---
3412.67 ± 1.539.67 ± 0.586.67 ± 0.58-
359.67 ± 0.58---
364.67 ± 0.58---
375.00 ± 1.73---
3910.67 ± 1.539.67 ± 0.584.33 ± 0.58-
419.67 ± 0.588.33 ± 0.583.00 ± 1.00-
426.67 ± 0.58---
434.67 ± 0.58---
444.67 ± 0.58---
455.33 ± 0.58---
468.33 ± 0.58---
476.67 ± 0.58---
484.67 ± 0.58---
514.67 ± 0.58---
523.33 ± 0.58---
532.33 ± 0.58---
542.33 ± 0.58---
553.00 ± 1.00---
564.33 ± 0.58---
574.67 ± 0.58---
584.67 ± 0.58---
595.67 ± 0.58---
605.33 ± 0.58---
626.67 ± 0.58---
634.33 ± 0.58---
642.67 ± 0.58---
653.67 ± 0.58---
664.33 ± 0.58---
675.33 ± 0.58---
685.67 ± 0.58---
695.67 ± 0.58---
703.33 ± 0.58---
714.33 ± 0.58---
723.33 ± 0.58---
734.67 ± 0.58---
744.67 ± 0.58---
752.33 ± 0.58---
763.67 ± 0.58---
812.33 ± 0.58---
824.67 ± 0.58---
833.33 ± 0.58---
842.33 ± 0.58---
867.67 ± 2.31---
875.00 ± 1.73---
894.67 ± 0.58---
902.67 ± 0.58---
913.67 ± 0.58---
922.33 ± 0.58---
931.67 ± 0.58---
943.67 ± 0.58---
954.67 ± 0.58---
963.33 ± 0.5810.33 ± 0.585.67 ± 0.58-
978.67 ± 0.5810.33 ± 0.585.67 ± 1.15-
984.67 ± 0.58---
995.67 ± 0.58---
1009.33 ± 0.584.33 ± 0.58--
ATB22.33 ± 0.58
Table
Table 7. Antimicrobial activity of samples against C. albicans in mm.
Table 7. Antimicrobial activity of samples against C. albicans in mm.
Sample No.50%25%12.5%6.25%
17.67 ± 0.58---
24.33 ± 0.58---
33.33 ± 0.58---
57.33 ± 0.58---
63.33 ± 0.58---
82.33 ± 0.58---
93.33 ± 0.58---
102.33 ± 0.58---
114.33 ± 0.58---
124.67 ± 0.58---
134.67 ± 0.58---
142.33 ± 0.58---
174.67 ± 0.58---
214.33 ± 0.58---
222.33 ± 0.58---
284.67 ± 0.58---
295.33 ± 0.58---
302.33 ± 0.58---
314.33 ± 0.58---
323.67 ± 0.58---
333.33 ± 0.58---
342.33 ± 0.58---
353.67 ± 0.58---
364.33 ± 0.58---
372.33 ± 0.58---
413.67 ± 0.58---
422.33 ± 0.58---
433.67 ± 0.58---
444.67 ± 0.58---
454.67 ± 0.58---
462.33 ± 0.58---
473.67 ± 0.58---
486.67 ± 0.58---
495.67 ± 0.58---
504.33 ± 0.58---
516.67 ± 0.58---
525.67 ± 0.58---
533.33 ± 0.58---
544.67 ± 0.58---
555.33 ± 0.58---
564.33 ± 0.58---
575.67 ± 0.58---
584.33 ± 0.58---
594.67 ± 0.58---
604.33 ± 0.58---
616.67 ± 0.58---
624.67 ± 0.58---
635.33 ± 0.58---
643.67 ± 0.58---
657.67 ± 0.58---
662.33 ± 0.58---
696.67 ± 0.58---
702.67 ± 0.58---
715.67 ± 0.58---
726.33 ± 0.58---
733.67 ± 0.58---
742.33 ± 0.58---
752.33 ± 0.58---
764.33 ± 0.58---
774.67 ± 0.58---
784.67 ± 0.58---
792.33 ± 0.58---
805.33 ± 0.58---
814.67 ± 1.15---
824.67 ± 0.58---
833.00 ± 1.00---
844.33 ± 0.58---
855.67 ± 0.58---
863.67 ± 0.58---
875.67 ± 0.58---
884.33 ± 0.58---
895.67 ± 0.58---
904.33 ± 0.58---
913.33 ± 0.58---
925.00 ± 1.00---
934.67 ± 0.58---
954.67 ± 0.58---
963.67 ± 0.58---
974.33 ± 0.58---
982.33 ± 0.58---
995.33 ± 0.58---
1004.67 ± 1.151.00 ± 1.00--
ATB20.33 ± 0.58
Table
Table 8. Antimicrobial activity of samples against C. glabrata in mm.
Table 8. Antimicrobial activity of samples against C. glabrata in mm.
Sample No.50%25%12.5%6.25%
17.33 ± 0.58---
22.33 ± 0.58---
33.67 ± 0.58---
42.33 ± 0.58---
52.33 ± 0.58---
73.33 ± 0.58---
112.33 ± 0.58---
137.33 ± 0.58---
142.33 ± 0.58---
174.67 ± 0.58---
183.33 ± 0.58---
214.67 ± 0.58---
222.67 ± 0.58---
234.33 ± 0.58---
243.33 ± 0.58---
296.67 ± 0.58---
306.33 ± 1.15---
318.33 ± 0.58---
328.67 ± 0.58---
344.33 ± 0.58---
355.67 ± 0.58---
364.67 ± 0.58---
374.67 ± 0.58---
382.33 ± 0.58---
396.67 ± 0.58---
407.67 ± 0.58---
414.67 ± 0.58---
423.33 ± 0.58---
434.67 ± 0.58---
445.67 ± 0.58---
456.67 ± 0.58---
463.33 ± 0.58---
475.67 ± 0.58---
485.67 ± 0.58---
496.67 ± 0.58---
504.67 ± 0.58---
513.33 ± 0.58---
524.67 ± 0.58---
535.33 ± 0.58---
546.33 ± 0.58---
553.67 ± 0.58---
562.33 ± 0.58---
574.67 ± 0.58---
584.00 ± 1.00---
594.67 ± 0.58---
607.67 ± 0.58---
624.33 ± 0.58---
654.67 ± 0.58---
663.33 ± 0.58---
679.33 ± 0.58---
686.67 ± 0.58---
695.33 ± 0.58---
705.33 ± 1.15---
718.33 ± 0.58---
724.67 ± 0.58---
736.67 ± 0.58---
743.33 ± 0.58---
764.67 ± 0.58---
773.33 ± 0.58---
782.33 ± 0.58---
794.33 ± 0.58---
804.67 ± 0.58---
813.67 ± 0.58---
824.67 ± 0.58---
833.33 ± 0.58---
845.33 ± 0.58---
852.67 ± 0.58---
864.33 ± 0.58---
873.33 ± 0.58---
882.67 ± 0.58---
894.67 ± 0.58---
902.33 ± 0.58---
912.33 ± 0.58---
921.67 ± 0.58---
936.67 ± 0.58---
943.33 ± 0.58---
954.67 ± 0.58---
962.33 ± 0.58---
983.67 ± 0.58---
992.33 ± 0.58---
10010.00 ± 2.003.33 ± 0.58--
ATB24.67 ± 0.58
Table
Table 9. Antimicrobial activity of samples against C. krusei in mm.
Table 9. Antimicrobial activity of samples against C. krusei in mm.
Sample No.50%25%12.5%6.25%
18.33 ± 0.58---
22.67 ± 1.15---
36.67 ± 0.58---
44.33 ± 0.58---
54.33 ± 0.58---
72.33 ± 0.58---
82.67 ± 0.58---
92.33 ± 0.58---
102.33 ± 0.58---
113.33 ± 0.58---
122.33 ± 0.58---
134.67 ± 0.58---
144.33 ± 0.58---
156.67 ± 0.58---
165.33 ± 0.58---
173.67 ± 0.58---
186.33 ± 0.58---
204.67 ± 0.58---
213.33 ± 0.58---
224.33 ± 0.58---
233.67 ± 0.58---
244.33 ± 0.58---
255.33 ± 0.58---
262.33 ± 0.58---
274.67 ± 0.58---
286.00 ± 1.00---
296.67 ± 0.58---
303.33 ± 0.58---
315.33 ± 0.58---
325.67 ± 0.58---
342.33 ± 0.58---
356.67 ± 0.58---
364.67 ± 0.58---
378.33 ± 0.58---
385.33 ± 0.58---
397.33 ± 0.58---
407.33 ± 0.58---
425.33 ± 0.58---
434.33 ± 0.58---
445.33 ± 0.58---
453.33 ± 0.58---
463.67 ± 0.58---
472.33 ± 0.58---
482.33 ± 0.58---
496.67 ± 0.58---
504.67 ± 0.58---
515.67 ± 0.58---
527.67 ± 0.58---
539.67 ± 0.58---
544.67 ± 0.58---
552.33 ± 0.58---
564.67 ± 0.58---
574.67 ± 0.58---
583.67 ± 0.58---
592.33 ± 0.58---
605.67 ± 0.58---
615.67 ± 0.58---
623.33 ± 0.58---
635.67 ± 0.58---
643.33 ± 0.58---
654.00 ± 1.00---
675.33 ± 0.58---
684.33 ± 0.58---
694.67 ± 0.58---
704.67 ± 0.58---
713.67 ± 0.58---
723.33 ± 0.58---
732.33 ± 0.58---
744.33 ± 0.58---
754.67 ± 0.58---
765.33 ± 0.58---
775.33 ± 0.58---
785.67 ± 0.58---
794.67 ± 0.58---
803.33 ± 0.58---
813.33 ± 0.58---
824.33 ± 0.58---
834.67 ± 0.58---
842.33 ± 0.58---
853.67 ± 0.58---
864.33 ± 0.58---
872.33 ± 0.58---
884.33 ± 0.58---
892.33 ± 0.58---
904.67 ± 0.58---
913.33 ± 0.58---
923.67 ± 0.58---
973.33 ± 0.58---
995.33 ± 0.58---
1000.67 ± 0.580.33 ± 0.58--
ATB22.33 ± 0.58
Table
Table 10. Antimicrobial activity of samples against C. tropicalis in mm.
Table 10. Antimicrobial activity of samples against C. tropicalis in mm.
Sample No.50%25%12.5%6.25%
17.33 ± 0.58---
23.33 ± 0.58---
34.67 ± 0.58---
43.33 ± 0.58---
54.67 ± 0.58---
63.67 ± 0.58---
75.33 ± 0.58---
83.67 ± 0.58---
94.67 ± 0.58---
102.33 ± 0.58---
113.33 ± 0.58---
122.33 ± 0.58---
133.33 ± 0.58---
142.33 ± 0.58---
155.33 ± 0.58---
164.67 ± 0.58---
173.33 ± 0.58---
182.33 ± 0.58---
194.67 ± 0.58---
203.67 ± 0.58---
214.00 ± 1.00---
222.33 ± 0.58---
234.67 ± 0.58---
243.33 ± 0.58---
254.33 ± 0.58---
263.33 ± 0.58---
283.67 ± 0.58---
294.67 ± 0.58---
304.33 ± 0.58---
312.67 ± 0.58---
324.67 ± 0.58---
334.67 ± 0.58---
345.67 ± 0.58---
352.33 ± 0.58---
363.33 ± 0.58---
373.33 ± 0.58---
384.67 ± 0.58---
394.67 ± 0.58---
404.67 ± 0.58---
412.33 ± 0.58---
435.33 ± 0.58---
444.67 ± 0.58---
453.67 ± 0.58---
463.33 ± 0.58---
474.33 ± 0.58---
483.67 ± 0.58---
494.67 ± 0.58---
503.67 ± 0.58---
512.33 ± 0.58---
525.67 ± 0.58---
533.67 ± 0.58---
542.33 ± 0.58---
557.33 ± 0.58---
564.33 ± 0.58---
574.67 ± 0.58---
585.33 ± 0.58---
595.67 ± 0.58---
607.67 ± 0.58---
624.33 ± 1.15---
633.33 ± 0.58---
645.33 ± 0.58---
654.67 ± 0.58---
665.67 ± 0.58---
674.67 ± 0.58---
687.33 ± 0.58---
694.67 ± 0.58---
704.67 ± 0.58---
714.33 ± 0.58---
725.67 ± 0.58---
734.67 ± 0.58---
742.67 ± 0.58---
753.67 ± 0.58---
793.67 ± 0.58---
803.67 ± 0.58---
812.33 ± 0.58---
823.00 ± 1.00---
832.33 ± 0.58---
842.33 ± 0.58---
853.33 ± 0.58---
862.33 ± 0.58---
874.33 ± 0.58---
883.67 ± 0.58---
894.33 ± 0.58---
903.33 ± 0.58---
913.67 ± 0.58---
922.67 ± 0.58---
934.67 ± 0.58---
953.67 ± 0.58---
964.00 ± 1.00---
975.67 ± 0.58---
983.67 ± 0.58---
993.33 ± 0.58---
10010.67 ± 1.156.33 ± 1.15--
ATB21.33 ± 0.58
Table
Table 11. Antioxidant activity of the honey (samples over 1000 TEAC).
Table 11. Antioxidant activity of the honey (samples over 1000 TEAC).
Sample No.% InhibitionTEAC
(μg Tx/1 g of Honey)		Type
6470.8 ± 1.22373.8 ± 37.7Buckwheat
9541.9 ± 0.71354.7 ± 23.5Honeydew
2839.7 ± 2.31280.8 ± 73.5Honeydew
3239.6 ± 0.21277.6 ± 6.4Honeydew
3439.2 ± 0.61264.7 ± 19.8Honeydew
3338.4 ± 0.11239.0 ± 3.2Honeydew
2737.4 ± 0.91206.8 ± 30.4Honeydew
3536.3 ± 1.71171.5 ± 53.6Chestnut
9436.2 ± 0.61168.2 ± 18.4Honeydew
6935.4 ± 1.81142.5 ± 58.5Buckwheat
9334.0 ± 0.31097.5 ± 9.6Manuka
9031.5 ± 1.51017.1 ± 48.9Mixed
8131.1 ± 0.81004.3 ± 25.3Mixed
Results of % inhibition and TEAC are presented as mean value ± SD.
Table
Table 12. Antioxidant activity according to the honey type.
Table 12. Antioxidant activity according to the honey type.
Type of HoneynDPPH Inhibition (%)TEAC (μg Tx/1 g of Honey)
RangeAverageRangeAverage
Acacia118.51–16.9611.71277.71–550.98378.54
Phacelia310.45–19.8116.14367.73–641.00532.76
Chestnut218.06–36.3127.19586.34–1171.46878.90
Mustard1-24.03-776.02
Linden914.26–24.2819.28460.96–785.67624.56
Manuka1-34.02-1097.52
Honeydew829.84–41.9437.78962.49–1354.721219.29
Fruit tree516.99–20.8719.27550.97–676.36624.92
Creamed rapeseed1-18.00-583.13
Buckwheat235.38–70.8353.111142.53–2373.851758.19
Rapeseed69.39–22.6816.95306.64–734.23541.33
Sunflower812.07–30.7821.08393.45–994.64682.79
Multifloral3510.95–25.2217.49354.87–814.60566.96
Mixed814.04–31.5423.77454.53–1017.15768.39
Table
Table 13. Details of the collected honey samples.
Table 13. Details of the collected honey samples.
CodeProducerTown/CountryLocality or MarketType
1beekeeperBratislava/SKtownmultifloral
2beekeeperBratislava/SKtownmultifloral
3beekeeperBratislava/SKtownmultifloral
4beekeeperBratislava/SKtownmultifloral
5beekeeperBratislava/SKtownmultifloral
6beekeeperTrnava/SKtownmultifloral
7beekeeperTrnava/SKtownmultifloral
8beekeeperTrnava/SLtownmultifloral
9beekeeperPrešov/SKtownmultifloral
10beekeeperPrešov/SKtownmultifloral
11beekeeperKošice/SKtownmultifloral
12beekeeperB. Bystrica/SKtownmultifloral
13beekeeperŽilina/SKtownmultifloral
14beekeeperNitra/SKtownacacia
15beekeeperNitra/SKtownlinden
16beekeeperZvolen/SKtownmultifloral
17beekeeperZvolen/SKtownmultifloral
18beekeeperZvolen/SKtownlinden
19beekeeperPezinok/SKtownmultifloral
20beekeeperPrievidza/SKtownmultifloral
21beekeeperPrievidza/SKtownmultifloral
22beekeeperKremnická lesy/SKforestlinden
23beekeeperB. Štiavnica/SKforestmultifloral
24beekeeperŠtrbské pleso/SKforestmultifloral
25beekeeperKraskovo/SKforestmultifloral
26beekeeperSabinov/SKforestmultifloral
27beekeeperSabinov/SKforesthoneydew
28beekeeperNitra/SKtownhoneydew
29beekeeperKremnica/SKforesthoneydew
30beekeeperPoltár/SKforestmultifloral
31beekeeperDetva/SKforestmultifloral
32beekeeperDetva/SKforesthoneydew
33beekeeperSenec/SKcountrysidehoneydew
34beekeeperLevoča/SKforesthoneydew
35beekeeperChoča/SKforestchestnut
36beekeeperOponice/SKforestchestnut
37beekeeperSenec/SKcountrysidemultifloral
38beekeeperSenec/SKcountrysidephacelia
39beekeeperChoča/SKcountrysiderapeseed
40beekeeperChoča/SKcountrysideacacia
41beekeeperHlohovec/SKcountrysidesunflower
42beekeeperOponice/SKcountrysidesunflower
43beekeeperOponice/SKcountrysideacacia
44beekeeperOponice/SKcountrysiderapeseed
45beekeeperŠala/SKcountrysiderapeseed
46beekeeperŠala/SKcountrysidesunflower
47beekeeperŠala/SKcountrysideacacia
48beekeeperLevice/SKcountrysideacacia
49beekeeperLevice/SKcountrysidesunflower
50beekeeperLevice/SKcountrysidephacelia
51beekeeperKrupina/SKcountrysidelinden
52beekeeperKrupina/SKcountrysideacacia
53beekeeperKrupina/SKcountrysidefruit trees
54beekeeperKysucké Nové mesto/SKtownacacia
55beekeeperKysucké Nové mesto/SKtownfruit trees
56beekeeperKysucké Nové mesto/SKtownsunflower
57beekeeperZáhorie/SKcountrysidelinden
58beekeeperZáhorie/SKcountrysiderapeseed
59beekeeperZáhorie/SKcountrysidesinapis
60beekeeperLiptov/SKforestmultifloral
61beekeeperHorná Streda/SKtownmultifloral
62beekeeperSenec/SKcountrysiderapeseed
63beekeeperSenec/SKcountrysidesunflower
64beekeeperNitra/SKcountrysidebuckwheat
65beekeeperKolíňany/SKcountrysidesunflower
66beekeeperKolíňany/SKcountrysidefruit trees
67beekeeperHlohovec/SKcountrysidemultifloral
68beekeeperHlohovec/SKcountrysidemultifloral
69beekeeperŠamorín/SKcountrysidebuckwheat
70beekeeperŠamorín/SKcountrysidelinden
71beekeeperŠamorín/SKcountrysidefruit trees
72beekeeperKysucké Nové mesto/SKcountrysidesunflower
73beekeeperKysucké Nové mesto/SKcountrysideacacia
74beekeeperKysucké Nové mesto/SKcountrysiderapeseed
75beekeeperKysucké Nové mesto/SKcountrysidelinden
76commerciallySKCBA—private labelmultifloral
77commerciallyEU—outside EULIDLmultifloral
78commerciallyEU—outside EUCOOP multifloral
79commerciallySKCOOP acacia
80commerciallySKCOOP multifloral
81commerciallySKCOOP multifloral
82commerciallySKCOOP multifloral
83commerciallySKCOOP—private labelmultifloral
84commerciallyEU—outside EUCOOP linden
85commerciallySKBillaacacia
86commerciallyEU—outside EUBilla—private labelmultifloral
87commerciallySKBillacreamed rapeseed
88commerciallySKTescolinden
89commerciallyEU—outside EUTescomultifloral
90commerciallyEU—outside EUTescomultifloral
91commerciallyEUKrajmultifloral
92commerciallyEU—outside EUKrajmultifloral
93commerciallyNew ZealandCeramelmanuka
94commerciallyTurkeyKauflandhoneydew
95commerciallyEU—outside EUKauflandhoneydew
96commerciallyEU—outside EUKauflandfruit trees
97commerciallySKKauflandacacia
98commerciallySKKauflandmultifloral
99commerciallyEUKauflandmultifloral
100beekeeperSKcountrysidephacelia
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Kačániová, M.; Borotová, P.; Galovičová, L.; Kunová, S.; Štefániková, J.; Kowalczewski, P.Ł.; Šedík, P. Antimicrobial and Antioxidant Activity of Different Honey Samples from Beekeepers and Commercial Producers. Antibiotics 2022, 11, 1163. https://doi.org/10.3390/antibiotics11091163

AMA Style

Kačániová M, Borotová P, Galovičová L, Kunová S, Štefániková J, Kowalczewski PŁ, Šedík P. Antimicrobial and Antioxidant Activity of Different Honey Samples from Beekeepers and Commercial Producers. Antibiotics. 2022; 11(9):1163. https://doi.org/10.3390/antibiotics11091163

Chicago/Turabian Style

Kačániová, Miroslava, Petra Borotová, Lucia Galovičová, Simona Kunová, Jana Štefániková, Przemysław Łukasz Kowalczewski, and Peter Šedík. 2022. "Antimicrobial and Antioxidant Activity of Different Honey Samples from Beekeepers and Commercial Producers" Antibiotics 11, no. 9: 1163. https://doi.org/10.3390/antibiotics11091163

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