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Honey meets acidity: a novel biopreservative approach against foodborne pathogens

  • Mohammad Melebari ORCID logo EMAIL logo , Ahmed G. Hegazi ORCID logo , Fayez M. Al Guthami , Mohamed F. Ramadan , Ahmed F.M. Al Gethami , Abeer M. Alghamdi , Fawzyah Obeedallah Albaldi , Nourah M. Almimoni , Tariq Alpakistany ORCID logo , Taher M. Taha , Alaa Ismail , Salud J. Serrano ORCID logo and Khaled S. Gazi
Published/Copyright: December 30, 2025
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Open Life Sciences
From the journal Open Life Sciences Volume 20 Issue 1

Abstract

Natural and bio-based alternatives to synthetic preservatives are gaining popularity in recent years. This study evaluates the antimicrobial potential and physicochemical properties of five types of honey vinegar. The investigation included 1,400 vinegar samples representing five types of formulations. Inhibition zone analysis was performed to assess the antibacterial activities of vinegar samples. All types of vinegars demonstrated antibacterial activity against at least one of the tested bacterial strains. Notably, honey-propolis and honey-black seed vinegars exhibited broad-spectrum antibacterial activity against all pathogens. Honey-garlic and honey-propolis vinegars contained the highest levels of total phenolics. These findings suggest that enriched honey vinegars, particularly those containing propolis or garlic, could be promising natural alternatives for antimicrobial applications. Their efficacy and rich bioactive profiles advocate their future use in functional health products and food preservation.

Keywords: honey vinegar; propolis-enriched vinegar; antibacterial activity; foodborne pathogens

1 Introduction

Antibacterial activity of vinegar has been reported against various food-borne bacterial pathogens [1]. A low acetic acid concentration (0.1 %) in vinegar could inhibit the growth of food-borne bacterial pathogens. Honey and vinegar have been widely used for wound cleaning and remedy [2], 3]. Honey, garlic, black seed (Nigella sativa), ginger, and propolis are rich in bioactive compounds, including organic acids, polyphenols, vitamins, flavonoids, and terpenoids. These biocompounds contribute to their multidimensional benefits (immune-modulating, antimicrobial, anti-inflammatory, and antioxidant effects), which help in promoting overall health [4], [5], [6], [7]. Functional foods, offering health benefits beyond basic nutrition, have gained considerable attention [8], 9]. Rich phytochemical profile and potential health-promoting properties of honey favor its in-depth exploration [2], 3], 10].

Vinegar, a fermented food product, is well known for its potential health benefits [11]. Its primary functions depend on the acetic acid content, which possesses antidiabetic, anti-glycemic, and polyphenol-based antioxidant properties [10]. The antimicrobial activity of vinegar facilitates its utilization in food safety and preservation [12]. The acetic acid content of the vinegar actively inhibits microbial growth [13].

The comparative efficacy of honey and vinegar combinations (honey, honey-garlic, honey black seed, honey ginger, and honey propolis vinegars) against Gram-positive and Gram-negative bacterial pathogens remains unexplored under standardized conditions. Therefore, the current study follows a disc diffusion method to evaluate the antimicrobial activity of honey and vinegar combinations against Gram-positive and Gram-negative bacterial strains.

2 Materials and methods

2.1 Vinegar samples

Five types of vinegar samples (n = 1,400) were used in this study, which included honey vinegar, honey garlic vinegar, honey black seed vinegar, honey ginger vinegar, and honey propolis vinegar. Vinegar samples were provided by Alnahal Aljwal Industry, Makkah, Saudi Arabia. Viniger contents were thoroughly mixed before the fermentation process, and all the ingredients and combinations were included at the fermentation stage. All samples were stored at 4 °C for later use after centrifugation to reduce the turbidity.

2.2 Bacterial Strains

A total of eight bacterial strains were used during the study, including three Gram-positive strains [Staphylococcus aureus (ATCC 25923), Bacillus cereus (ATCC 11778), and Listeria monocytogenes (ATCC 19115)] and five Gram-negative strains [Vibrio parahaemolyticus (ATCC 17802), Escherichia coli O157 (ATCC 43888), E. coli (ATCC 25922), Pseudomonas aeruginosa (NCTC 10662), and Salmonella typhimurium (ATCC 14028)] to evaluate the antibacterial efficacy of different vinegar combinations. All the bacterial strains were provided by the Microbiology Laboratory, Faculty of Science, Al-Baha University, Saudi Arabia. The bacterial cultures were stored at −80 °C in a glycerol-containing (20 %) tryptic soy broth (TSB) until used. The standardization and microbial inoculation were performed by following the methodology of Melebari [14].

2.3 Disk diffusion assay

The disk diffusion (Kirby-Bauer) assay was carried out to assess the antibacterial efficacy of vinegar combinations [15], 16]. TSB was used to culture bacterial isolates, followed by incubation (37 °C) for 18 h. The bacterial suspensions (1–5 × 108 CFU/mL) of the studied isolates were prepared and spread over Muller-Hinton Agar (MHA) plates. The vinegar-loaded disks were simultaneously prepared as well. The procedure involved the making of a six-layer disk by folding Whatman No. 1 filter papers three times, followed by punching with a paper puncher. The disks were autoclaved and oven-dried at 70 °C. Vinegar samples were aseptically loaded (50 µL/disk) on the sterilized dried disks. The loaded disks were again dried (60 °C) for 2 h. Some volatile compounds might be lost during the drying process, which could probably undermine the full antibacterial potential of the liquid vinegar. The loaded dried disks were added to the inoculated MHA plates and incubated at 37 °C. Inhibition zone diameters (mm) were measured after 6, 12, 18, and 24 h. The treatments were performed in triplicate, and their averages were calculated (Figures 1 and 2). Ampicillin and streptomycin served as the positive antibiotic controls for Gram-positive and Gram-negative bacteria, respectively.

Figure 1: Effect of various honey and vinegar combinations on Gram-positive bacteria over time. Data denotes comparative antibacterial impacts of honey and vinegar, honey-garlic vinegar, honey-ginger vinegar, honey-black seeds vinegar, honey-propolis vinegar, and Ampicillin (control group) treatments. Data collected at 6, 12, 18, and 24 h post-treatment illustrate differential responses of each treatment. Data are presented as mean ± standard deviation (n = 3 replicates). A t-test was performed to establish statistical significance at p < 0.05.
Figure 1:

Effect of various honey and vinegar combinations on Gram-positive bacteria over time. Data denotes comparative antibacterial impacts of honey and vinegar, honey-garlic vinegar, honey-ginger vinegar, honey-black seeds vinegar, honey-propolis vinegar, and Ampicillin (control group) treatments. Data collected at 6, 12, 18, and 24 h post-treatment illustrate differential responses of each treatment. Data are presented as mean ± standard deviation (n = 3 replicates). A t-test was performed to establish statistical significance at p < 0.05.

Figure 2: Effect of various honey and vinegar combinations on Gram-negative bacteria over time. Data denotes comparative antibacterial impacts of honey and vinegar, honey-garlic vinegar, honey-ginger vinegar, honey-black seeds vinegar, honey-propolis vinegar, and Ampicillin (control group) treatments. Data collected at 6, 12, 18, and 24 h post-treatment illustrate differential responses of each treatment. Data are presented as mean ± standard deviation (n = 3 replicates). A t-test was performed to establish statistical significance at p < 0.05.
Figure 2:

Effect of various honey and vinegar combinations on Gram-negative bacteria over time. Data denotes comparative antibacterial impacts of honey and vinegar, honey-garlic vinegar, honey-ginger vinegar, honey-black seeds vinegar, honey-propolis vinegar, and Ampicillin (control group) treatments. Data collected at 6, 12, 18, and 24 h post-treatment illustrate differential responses of each treatment. Data are presented as mean ± standard deviation (n = 3 replicates). A t-test was performed to establish statistical significance at p < 0.05.

2.4 Physicochemical properties

A pH meter (InoLab 720, WTW GmbH, Germany) was used to measure the pH. The acidity is expressed in grams of acetic acid per liter. NaOH (0.1 mol/L) was used to calculate the percentage of acetic acid [17], 18]. A refractometer assisted in assessing the total acidity in equivalence to acetic acid [18], [19], [20]. The Brix values were measured, whereas the residual alcohol content percentage represented the ethanol volume remaining in the vinegar after the acetic acid fermentation [20]. The total dry matter extract refers to all the substances under particular conditions as described by I.O.V [20]. The total dry matter percentage was calculated by following the Association of Official Analytical Chemists methods [18]. A refractometer (ATOGO, China) was used to measure total soluble solids according to Hidalgo [21]. Vinegar ash represents the evaporation residue’s incineration products within a known vinegar volume [18], 22], 23]. Mineral analysis was carried out by following previous studies [22], [23], [24]. The vinegar titratable acidity was calculated as the acetic acid percentage (%) [18]. The turbidity values of different types of vinegars were detected using a turbidimeter and expressed as NTU (Nephelometric Turbidity Unit) [24], 25].

2.5 Bioactive value

The total phenolic content (TPC) was measured spectrophotometrically by adopting the Folin-Ciocalteu method [26]. Mixture absorbance was determined and expressed as µg gallic acid equivalents per mL of vinegar samples (GAE/mL) [27]. The total flavonoid content (TFC) was also assessed spectrophotometrically at 510 nm, and expressed as Quercetin Equivalent per mL of vinegar (µg QE/mL) [28].

2.6 Statistical analysis

R software (4.4.3) was used for data processing and visual output. The data were statistically analyzed (two-way ANOVA) to differentiate the impacts of various treatments on bacterial strains based on the size of the inhibition zones. Inhibition zone diameter (mm) served as the response variable, whereas the treatment conditions and bacterial strains were considered as the controlled effects. Tukey’s Honestly Significant Difference (HSD) test compared the means at a significance level of 5 % (α = 0.05), and established significant differences among treatment groups based on their antimicrobial performance. Bar charts display the average inhibition zones and corresponding standard deviations (SD). The error bars represent variability across replicates at different experimental periods, thus revealing technical and biological fluctuations during the study [2].

3 Results

Inhibition zone diameters were monitored at 6, 12, 18, and 24 h after the treatment to estimate the antibacterial efficacy of the studied vinegar combinations against eight bacterial species (V. parahaemolyticus, S. typhi, P. aeruginosa, E. coli O157:H7, E. coli, S. aureus, B. cereus, and S. monocytogenes). These strains represented a diverse set of pathogens (foodborne and environmental), which differ in their cell wall characteristics and antibiotic resistance profiles.

The disk diffusion method was followed to assess the antibacterial activity of different honey and vinegar combinations. Inhibition zone diameter served as the primary indicator of antibacterial efficacy. Two-way ANOVA indicated inhibition zone-based differential impacts of various treatments on the tested bacterial strains. The results demonstrated that honey vinegar combinations significantly suppressed bacterial growth (p < 0.001), regardless of the strain. However, the interaction between bacterial strain and treatment type remained non-significant, indicating consistent effectiveness of all treatments against different pathogens.

Honey propolis vinegar treatment consistently produced significantly larger inhibition zones as compared to other combinations. It was followed by honey-ginger vinegar and honey black seed vinegar, which exhibited moderate bacterial inhibition. Contrarily, honey vinegar and honey-garlic vinegar demonstrated weak antibacterial effects against both Gram-positive and Gram-negative strains. Thus, these combinations consistently appeared in the least effective statistical groups (Figures 1 and 2).

Vinegar formulations displayed differential antimicrobial efficacy against Gram-positive bacteria as compared to the control treatment. Honey black seed vinegar (0.1042) and honey propolis vinegar (0.1042) shared similar antimicrobial activity with comparable bacterial inhibition to streptomycin (p < 0.05). Contrarily, the antibacterial activities of honey ginger vinegar (0.0028), honey garlic vinegar (0.00048), and honey vinegar (0.0309) were significantly lower than other treatments (p < 0.05). These findings depict higher antibacterial potential of certain honey-enriched vinegars (honey black seed and honey propolis vinegars) than conventional synthetic antibiotics, which highlights their potential as natural alternatives for microbial control.

Similarly, vinegar formulations demonstrated differential antimicrobial activity against Gram-negative bacteria in comparison to the control treatment. Honey propolis vinegar (p = 0.1014) exhibited significantly higher antibacterial potential with a comparable inhibition zone diameter to the control treatment (p < 0.05). Contrarily, the antibacterial efficacies of honey black seed vinegar (p = 0.0013), honey ginger vinegar (p = 0.0016), honey garlic vinegar (p = 3.78 × 10−8), and honey vinegar (p = 2.50 × 10−10) were significantly lower than the control treatment. The results present honey propolis vinegar as a natural alternative for microbial control.

Overall, the results validated the hypothesis that the addition of certain bioactive ingredients could enhance the antimicrobial efficiency of honey vinegar. A significantly high antibacterial activity of propolis-containing vinegar could be attributed to its high phenolic acid and flavonoid content. These compounds collectively contribute to increased activity of enzymatic compounds and natural acidity in the fermented honey, which could yield broad-spectrum antibacterial properties.

Figure 3 presents the physicochemical properties of vinegars (1,400 samples), which reveal significantly different vinegar qualities. Vinegar samples’ pH levels varied from 3.40 ± 0.02 (honey propolis vinegar) to 3.54 ± 0.01 (honey ginger vinegar). Generally, the highest pH was observed in honey propolis vinegar (3.40 ± 0.02), whereas the lowest pH was noted in honey ginger vinegar (3.54 ± 0.01). The total acidity levels were expressed in equivalence to acetic acid, which correlated with vinegar samples’ pH values (Figure 3). Honey propolis vinegar demonstrated the highest total acidity (1.22 ± 0.25 g/L), whereas the lowest acidity was noted in honey ginger vinegar (1.68 ± 0.06 g/L). The results depicted a wide range of vinegar °Brix values (3.34 ± 0.09 to 4.12 ± 0.03 g/cm3). The °Brix values indicated the percentage of total soluble solids (sugar, proteins, and salts) in an aqueous sample (Figure 3). Vinegar turbidity represents the presence of suspended solids in the liquid medium. The turbidity levels of different vinegars varied between 18.6 ± 0.57 and 215.0 ± 16.3 NTU. The honey propolis vinegar displayed the highest turbidity (215.0 ± 16.3), whereas the lowest value of 18.6 ± 0.57 was noted in honey vinegar (Figure 4).

Figure 3: Physicochemical properties of tested vinegars.
Figure 3:

Physicochemical properties of tested vinegars.

Figure 4: Bioactive properties of tested vinegars.
Figure 4:

Bioactive properties of tested vinegars.

The total phenolic and flavonoid contents (TPCs and TFCs) of vinegar samples and their bioactive properties are summarized in Figure 4. The bioactive properties of vinegar samples varied among different vinegar types. The means for total phenolic content ranged from 235.81 ± 14.71 µg GAE/mL (honey vinegar) to 288.19 ± 11.14 µg GAE/mL (honey propolis vinegar). The TFC ranged from 144.49 ± 0.76 µg QE/mL to 479.05 ± 2.37 µg QE/mL for honey vinegar and honey propolis vinegar, respectively (Figure 4). Figures 5 and 6 depict the mineral contents in the tested vinegar samples. The level of iron remained highest in all vinegars, followed by sodium, potassium, calcium, and magnesium. Among other minerals (copper, manganese, cobalt, zinc, selenium, chromium, and nickel), the level of manganese remained the highest in all vinegars. Overall, the results established a correlation between physicochemical parameters of vinegar samples and their antibacterial activities (Table 1).

Figure 5: Sodium, potassium, calcium, magnesium, and iron levels in different vinegars.
Figure 5:

Sodium, potassium, calcium, magnesium, and iron levels in different vinegars.

Figure 6: Copper, manganese, cobalt, zinc, selenium, chromium, and nickel levels in different vinegars.
Figure 6:

Copper, manganese, cobalt, zinc, selenium, chromium, and nickel levels in different vinegars.

Table 1:

Pearson correlation between physicochemical parameters of honey vinegars and their mean antibacterial activity (mean inhibition zone across all bacteria).

Parameter r (correlation) p-value Significance
pH −0.83 0.083 Not significant (trend, negative)
Acidity −0.82 0.088 Not significant (trend, negative)
Brix +0.98 0.0022 Significant
Turbidity +0.997 0.00023 Highly significant
TPC (phenolics) +0.96 0.0081 Significant
TFC (flavonoids) +0.996 0.00028 Highly significant

  1. P-value < 0.05 (significant); P-value < 0.001 (Highly significant).

4 Discussion

The study revealed intriguing antibacterial potential of different types of honey vinegars (honey, honey garlic, honey ginger, honey black seeds, and honey propolis). Honey vinegar combined with different additives demonstrated varying antibacterial activities against the tested bacterial strains. A synergic antibacterial efficacy of honey garlic and honey ginger mixtures has been reported against K. pneumoniae and P. aeruginosa, respectively [24]. The evaluated honey-based vinegars’ antibacterial efficacy differed depending on the type of bacterium and the length of incubation (Figure 1). Throughout the 24-h period, the growth inhibition zones stayed near the negative control, indicating that none of the honey vinegars significantly inhibited S. aureus. On the other hand, L. monocytogenes was moderately inhibited by all treatments, with honey propolis vinegar consistently producing greater inhibition zones than the other formulations. Notably, honey garlic vinegar and honey propolis vinegar demonstrated the strongest action against B. cereus, outperforming the control antibiotic (ampicillin) with distinct inhibition zones that reached up to 16–17 mm after 18 h. These results suggest that certain honey-based vinegars, especially those enriched with garlic and propolis, possess potent antibacterial properties that may be species-specific, with B. cereus appearing particularly susceptible. These results are in partial agreement with our previous study on date-based vinegars [29]. Later on, the antibacterial activity was not observed, and the strain grew normally. Mahmoud et al. [30] have reported apple vinegar solution-based prevention of B. cereus growth and biofilm formation. The biofilm inhibition capability of vinegar was positively associated with the antibacterial activity of the acetic acid. These results are in line with the findings of Halstead et al. [31]. They reported that acetic acid restricted the biofilm formation. Acetic acid content is often high (3–5%) in fermented vinegars. Ng et al. (2023) have also demonstrated acetic acid (0.31 %)-based inhibition of biofilm formation in P. aeruginosa [32]. Moreover, low concentrations of apple cider vinegar could inhibit the growth of C albicans, E. coli, and S. aureus [33]. Natural products, including ginger and honey, have gained significant attention for managing bacterial infections [34], 35].

The antibacterial effects of honey-based vinegars demonstrated variable patterns against Gram-negative bacteria, including Vibrio sp., P. aeruginosa, S. typhi, and E. coli strains (Figure 2). The strongest inhibitory action was repeatedly demonstrated by honey propolis vinegar, especially against Vibrio sp. and E. coli O157, where inhibition zones reached up to 14–16 mm after 24 h, matching or surpassing the control antibiotic (streptomycin). This emphasizes the role of phenolic chemicals derived from propolis, which are known to interfere with quorum sensing and break down bacterial membranes. Significant inhibitory effects were also demonstrated by honey garlic vinegar, particularly against P. aeruginosa and E. coli O157, where inhibition zones were maintained between 8 and 12 mm during various incubation durations. Allicin and other sulfur-containing chemicals found in garlic are responsible for this activity. They interfere with thiol-containing enzymes to provide a broad-spectrum antibacterial effect.

On the other hand, the majority of the organisms examined showed moderate inhibition from honey black seed vinegar and honey ginger vinegar, with inhibition zones often falling between 6 and 10 mm. Their activity outperformed \ honey vinegar, although being less effective than honey garlic and honey propolis venigers. This suggests that adding bioactive spices to honey vinegar improves their antibacterial efficiency. It’s interesting to note that overall, S. typhi was less responsive than E. coli. Inhibition zones for Vibrio sp. and E. coli are rarely larger than 8–10 mm. The pathogen’s strong outer membrane and efflux pump mechanisms, which frequently lessen vulnerability to natural antimicrobials, may be the cause of this resistance. All organisms were consistently inhibited by the control antibiotic, streptomycin, but in certain instances especially E. coli. The effects of honey propolis vinegar on Vibrio sp. and E. coli O157 were either similar or more pronounced.

These results highlight the potential of natural formulations, such as honey-propolis and honey-garlic vinegars, as supplementary or effective substitutes for traditional antibiotics, particularly when used against Gram-negative bacteria that are resistant to several drugs. The potential public health significance of these natural antimicrobials is further highlighted by their proven efficacy against E. coli O157, a significant foodborne pathogen.

These results are in agreement with previous findings where antimicrobial effects of vinegar were noted against various microorganisms. High acidity, microbial metabolites, phenolic substances, and organic acids in vinegar composition contributed to its antimicrobial properties [2], 35], 36]. All five vinegars effectively inhibited E. coli growth after 6 h of incubation. Vinegars disturbed E. coli O157 growth more than E. coli sp. up to 6 h, except pure honey vinegar [1]. During this study, the antibacterial activity of honey vinegar was comparatively lower than other vinegars. Similarly, honey garlic vinegar demonstrated antibacterial efficacy against only three strains (B. cereus, S. typhi, and E. coli sp.) as compared to the other three types of vinegars [35]. These results are in agreement with previous studies, which reported varying antibacterial potential of apple vinegar against different bacteria, such as V. colerae, B. cereus, E. coli, and Candida albicans [36], 37]. Overall, vinegars exerted antimicrobial impacts on various Gram-positive and Gram-negative microorganisms [1]. Antimicrobial effectiveness of several types of vinegars (rice berry, honey, wood, pomegranate, grape, blueberry, apple, and industrial vinegars) has been investigated as well [29], 38], 39].

Figure 3 illustrates the physicochemical properties of the studied vinegars [n = 1,400]. Vinegar samples largely shared similar physicochemical profiles, and vinegar formulations were largely comparable with minor variations. Major differences were observed in cation concentrations and total flavonoid content, which might impact antibacterial potential. Turbidity-associated differences are generally of less biological importance. Despite similarities, the measured parameters provided useful information regarding vinegar quality and bioactive compounds contributing to the antimicrobial efficacy of vinegars. pH levels varied among vinegars from 3.40 ± 0.02 (Honey propolis vinegar) to 3.54 ± 0.01 (honey ginger vinegar). The highest pH was observed in honey propolis (3.40 ± 0.02), whereas the lowest pH was noted in honey ginger vinegar (3.54 ± 0.01). Ozturk et al. [39] have also observed similar results where pH values significantly varied with the fruit source and fermentation method, and ranged from 2.84 ± 0.01 to 3.82 ± 0.04. The lowest pH was recorded in the Gala cultivar vinegar [39]. Another study has reported a pH range of 2.84 ± 0.01 to 3.82 ± 0.04 in Moroccan apple vinegars. They emphasized the influence of raw material on vinegar acidity [35].

The total acidity levels were expressed in equivalence to acetic acid, which correlated with the pH values of the vinegar samples. Honey propolis vinegar had the highest total acidity (1.22 ± 0.25 g/L), whereas the lowest acidity was noted in honey ginger vinegar (1.68 ± 0.06 g/L). This could be due to the close relationship of vinegar acidity to the honey sugar content. It directly influences the ethanol and acetic acid contents of the vinegar that is obtained after the fermentation (alcoholic and acetic). Similar results have been documented in previous studies [40], [41], [42]. Özdemir [43] demonstrated strong antioxidant and antibacterial effects of red fruit vinegar postbiotic solutions, which are produced through traditional fermentation. Their antimicrobial efficacy is largely attributed to phenolic and acid content.

During the current study, vinegars’ °Brix values widely varied from 3.34 ± 0.09 to 4.12 ± 0.03 g/cm3. It indicates the percentage of total soluble solids (sugar, proteins, and salts) in an aqueous sample [44], [45], [46]. The suspended solids in the liquid medium contribute to the vinegar turbidity [43]. The turbidity levels varied among different vinegars and ranged between 18.6 ± 0.57 to 215.0 ± 16.3 NTU. Cosmulescu et al. have reported that vinegar’s turbidity spoilage could sharply tamper with its quality. The highest turbidity was noted in the honey propolis vinegar (215.0 ± 16.3), whereas the lowest value of 18.6 ± 0.57 was observed in honey vinegar. Propolis-added hawthorn vinegar is known to provide functionality in salads [47]. Generally, vinegars’ pH remained above 3.00, which is in line with the literature. Previous studies have correlated total acidity levels with their pH values, which is in accordance with the Codex Alimentarius Commission [48], 49].

The total phenolic and flavonoid contents (TPCs and TFCs) of vinegars are summarized in Figure 4. Bioactive properties varied among different types of vinegars [50]. Figure 4 depicts the profile of the bioactive compounds. The means of total phenolic content ranged from 235.81 ± 14.71 µg GAE/mL (honey vinegar) to 288.19 ± 11.14 µg GAE/mL (honey propolis vinegar). TFC ranged from 144.49 ± 0.76 µg QE/mL (honey vinegar) to 479.05 ± 2.37 µg QE/mL (honey propolis vinegar) (Figure 4). Mineral contents in the studied vinegar types are presented in Figures 5 and 6. Iron had the highest level in all vinegars followed by sodium, potassium, calcium, and magnesium (Figure 5). Among other minerals (copper, manganese, cobalt, zinc, selenium, chromium, and nickel), the level of manganese remained highest in all vinegars (Figure 6). Similar findings have been reported in previous studies [51], [52], [53], [54]. The results of this study revealed a correlation between antibacterial activity and physicochemical parameters of honey vinegars, which is in line with the findings of El Abdali et al. [35]. Moreover, the study revealed strong positive correlations of Brix, Turbidity, TPC, and TFC with antibacterial activity of the investigated vinegar types (p < 0.01). Contrarily, non-significantly negative correlations were noted between pH and acidity (p > 0.05). High acidity at a lower pH was found to increase the activity. Turbidity and flavonoid content (TFC) emerged as the most influential factors, followed by phenolic content (TPC).

5 Conclusions

The findings of this study provide valuable insight into the antimicrobial properties of five honey vinegar formulations produced in Saudi Arabia. The results highlighted that enriched honey vinegars are highly potent to serve as natural antimicrobial agents in public health strategies, future therapies, and food preservation. Statistical analysis further established the credibility of results for future in vivo and shelf-life investigations. Notably, the vinegar treatments effectively exerted antibacterial effects within the first 6 h against common foodborne pathogens. Therefore, the incorporation of these vinegars into the daily fresh diet could enhance protection against microbial infections and support overall health.

Corresponding author: Mohammad Melebari, Department of Biology, Faculty of Science, Al-Baha University, Al-Baha 65431-65779, Saudi Arabia, E-mail:

Acknowledgments

The authors extend their appreciation to The National Research Centre, Egypt, and Al Guthami Company.

  1. Funding information: The paper is funded by the National Research Centre, Egypt, and Al Guthami Company, Saudi Arabia.

  2. Author contribution: Conceptualization, M.M., A.G.H., and A.F.M.G.; methodology, M.F.A.R., K.S.G., N.A., T.A., F.O.A., and A.M.A.; data curation, A.F.G., F.A., and T.M.T.; writing – original draft preparation, A.G.H., K.S.G., and S.S.J.; writing, review, and editing, M.M., K.S.G., and T.M.T.; supervision, F.M.G. All authors have read and agreed to the published version of the manuscript.

  3. Conflict of interest: Authors state no conflict of interest.

  4. Data availability statement: The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Supplementary Material

This article contains supplementary material (https://doi.org/10.1515/biol-2025-1225).

Received: 2025-06-11
Accepted: 2025-10-29
Published Online: 2025-12-30

© 2025 the author(s), published by De Gruyter, Berlin/Boston

This work is licensed under the Creative Commons Attribution 4.0 International License.

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  166. Revisiting the link between NADPH oxidase p22phox C242T polymorphism and ischemic stroke risk: an updated meta-analysis
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  168. Overexpression of HE4/WFDC2 gene in mice leads to keratitis and corneal opacity
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  175. The complete mitochondrial genome analysis of Haemaphysalis hystricis Supino, 1897 (Ixodida: Ixodidae) and its phylogenetic implications
  176. Epidemiological characteristics and risk factors analysis of multidrug-resistant tuberculosis among tuberculosis population in Huzhou City, Eastern China
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  179. Diversity and environmental drivers of Suillus communities in Pinus sylvestris var. mongolica forests of Inner Mongolia
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  205. Using plant electrical signals of water hyacinth (Eichhornia crassipes) for water pollution monitoring
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  211. Review on role of honey in disease prevention and treatment through modulation of biological activities
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  228. Preparation and characterization of lemongrass oil nanoemulsion: Antimicrobial, antibiofilm, antioxidant, and anticancer activities
  229. Fluorescent detection of sialic acid–binding lectins using functionalized quantum dots in ELISA format
  230. Smart tectorigenin-loaded ZnO hydrogel nanocomposites for targeted wound healing: synthesis, characterization, and biological evaluation
  231. Corrigendum
  232. Corrigendum to “Utilization of convolutional neural networks to analyze microscopic images for high-throughput screening of mesenchymal stem cells”
  233. Corrigendum to “Effects of Ire1 gene on virulence and pathogenicity of Candida albicans”
  234. Retraction
  235. Retraction of “Down-regulation of miR-539 indicates poor prognosis in patients with pancreatic cancer”
https://doi.org/10.1515/biol-2025-1225
Keywords for this article
honey vinegar; propolis-enriched vinegar; antibacterial activity; foodborne pathogens
Creative Commons
BY 4.0

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  234. Retraction
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