Moreover, another study demonstrated that a 10% concentration of Manuka honey was able to inhibit the formation of a biofilm of oral bacteria such as Streptococcus mutans, suggesting that honey might be able to reduce oral pathogens within dental plaque [77]. On the other hand, light-colour honeys such as clover honey (pasture honey) and acacia or rapeseed honey showed to be less potent as antibacterial agent with MIC higher than 25–50% (w/v). Honey is antibacterial and can prevent growth of most types of bacteria. As shown in Figure 1, the different active components in honey have been isolated by neutralizing each one individually and observing the effect on its antimicrobial activity. Six types of honey varieties were studied by Lusby et al., to investigate the antibacterial activity against 13 species of bacteria and one yeast species [34]. In contrast, Agbaje et al., reported that 100% honey might not proffer a total solution to the current problems facing bacterial chemotherapy when compared to 0.2% ciprofloxacin and 2.5% tetracycline [80]. There are different varieties of lemon. In other methods, honey is incorporated into the nutrient agar or into the nutrient broth in which the bacterial culture is grown. Sign up here as a reviewer to help fast-track new submissions. By using a series of different concentrations of honey within the broth or agar, it is possible to determine the minimum inhibitory concentration (MIC) for each type of honey studied [10]. This discovery has provoked an increase in the number of studies that have investigated the effect of substances other than peroxide activity. The most commonly used bacterial susceptibility assay is a broth micro- or macrodilution assay. Honey is one such product that used to be widely used to combat bacteria. These phenolic acids included protocatechuic acid, p-hydroxybenzoic acid, caffeic acid, chlorogenic acid, vanillic acid, p-coumaric acid, and benzoic acid. Viper’s bugloss and Heather honey have also been studied and shown to have a much lower phenolic acid content, ranging between 132.17 ± 0.05 and 727.77 ± 0.23 mg/Kg [55]. including antibacterial, antifungal, antidiabetic, anticancer and antiviral activities [7]. This experiment compares the effect of different types of honey on bacteria growing on agar plates. Honey has been shown to have an antibacterial effect on bacteria commonly present in wounds, such as methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci (VRE). Furthermore, it is possible to differentiate honey into two main types: floral honey that is made from the nectar of blossoms (blossom honey) and honeydew honey is prepared from the secretions of living parts of plants or the excretions of plant-sucking insects [8, 9]. The moisture content of honey can also vary between different honey varieties and can be affected by climate, season, and moisture content of the original plant nectar. Many factors have been shown to contribute to the antibacterial activity of honey, such as its high viscosity, mostly due to a high sugar concentration and low water content, which helps to provide a protective barrier to prevent infection. Protein content in honey is very low and ranges between 0.1 and 0.5%. The aim of this experiment was to find a solution that may help control the resistance of bacteria to conventional antibiotics. E. coli: Escherichia coli; EPHI: Ethiopian Public Health Institute; EHNRI: Ethiopian Health and Nutrition Research Institute; Lab: Laboratory; MRSA: Methicillin Resistant Staphylococcus aureus; MSA: Mannitol Salt Agar; Staph: Staphylococcus. antibacterial applicant (Miorin et al., 2003). White et al., reported that gluconic acid is an effective antibacterial factor produced as a result of glucose oxidation by endogenous glucose oxidase [30]. Scholars have determined the level of hydrogen peroxide in a large number of honey samples as summarised in Table 2 [41–46]. Many aspects of the antibacterial properties of honey have been reviewed and the growth of different bacteria has been tested in the presence of different concentrations of honey [4, 66, 70]. Different proteins have been detected in different honey varieties, predominantly related to different types of honeybees or different types of plants/flowers [27]; however, a group of major royal jelly proteins are shared by all honeybees. Honey contains antioxidants and flavonoids that may function as antibacterial agents. In this study, E. coli showed more susceptibility to inhibition by the honey than P. aeruginosa [76]. This demonstrates the wide range of compounds that could contribute to the antibacterial properties of honey. It is important to note that the level of hydrogen peroxide in honey is also determined by the presence and action of catalase. After analysis, the scientists concluded that the vast majority of honey's antibacterial properties come from that protein. Propolis, a flavonoid-rich product of honey comb, exhibits antibacterial and anti-inflammatory properties (Bosio et al., 2000) which is very powerful *Corresponding author. They tested the growth of bacteria in media which contained different concentrations of honey, namely, 10%, 20%, 30%, 40%, and 50% (w/v). There are several other methods that have been used to measure the antibacterial activity of honey. Water activity is a measure of the unbound water molecules in food; the less the unbound water, the harder it is for bacteria to grow in foods. Generation of hydrogen peroxide, bee defensin-1, high osmolarity and low value of pH seems to be crucial for its antimicrobial potential. Hydrogen peroxide (H2O2) is an important oxidizing and sanitizing agent [40]. Bees collect many materials to produce honey, including nectar, volatiles essential oils, pollen, and propolis, and these various botanical origins will also affect the composition of honey [11]. This means that new strategies are needed to prevent and treat infectious disease. According to some studies, honey has been shown to possess a high level of phenolic compounds which might contribute to its antibacterial activity. The researcher identifies the antibacterial properties of honey with isolated compounds derived from honey, selected antibiotics. These enzymes have been studied and have been shown to contain similar properties of antibiotics. This study aims to investigate antibacterial activity of five varieties of Malaysian honey (three monofloral; acacia, gelam and pineapple, and two polyfloral; kelulut and tualang) against Staphylococcus aureus, Bacillus cereus, Escherichia coli, and Pseudomonas … This experiment compares the effect of different types of honey on bacteria growing on agar plates. of gentamicin and three kinds of pure honey obtained from Ibadan and Abeokuta in south west Nigeria, using undiluted and fresh aqueous dilutions of 1 : 2, 1 : 4, and 1 : 6 in an agar diffusion method. Several research studies have investigated honey and its effect on various species of bacteria (Table 3). It is important to note, however, that in this assay the effective antibacterial concentration can be lower than the concentration applied to the agar due to honey’s dilution during diffusion [10]. As early as the 1990s, phenolic acids and flavonoids were recognised as important components of the antibacterial substances in honey [52]. Explanation of the antibacterial activity of honey Osmotic effect Honey is a saturated or super-saturated solution of sugars, the water content usually being only 15-21% by weight124. Methylglyoxal (MGO; CH3-CO-CH=O or C3H4O2) is also an important constituent of honey that has recently been shown to contribute to its antibacterial activity with a minimum inhibition concentration (MIC) of 1.1 mM when tested against E. coli and S. aureus [60]. The bacterial growth (change in turbidity) is assessed spectrophotometrically. Moreover, in some cases, these products/compounds can be used in combination with antibiotics to enhance their activity. A. Adeniyi, and G. B. Mahady, “Use of natural products to combat multidrug-resistant bacteria,” in, G. B. Mahady, Y. Huang, B. J. Doyle, and T. Locklear, “Natural products as antibacterial agents,”, P. Molan, “Honey: antimicrobial actions and role in disease management,” in, M. S. Butler and A. D. Buss, “Natural products—the future scaffolds for novel antibiotics?”, M. D. Mandal and S. Mandal, “Honey: its medicinal property and antibacterial activity,”, J. M. Alvarez-Suarez, S. Tulipani, D. Díaz et al., “Antioxidant and antimicrobial capacity of several monofloral Cuban honeys and their correlation with color, polyphenol content and other chemical compounds,”, M. L. Sanz, M. Gonzalez, C. de Lorenzo, J. Sanz, and I. Martı́nez-Castro, “A contribution to the differentiation between nectar honey and honeydew honey,”, A. Bentabol Manzanares, Z. H. García, B. R. Galdón, E. R. Rodríguez, and C. D. Romero, “Differentiation of blossom and honeydew honeys using multivariate analysis on the physicochemical parameters and sugar composition,”, P. C. Molan, “The antibacterial activity of honey,”, P. J. Torley, R. P. G. Rutgers, B. D’Arcy, and B. R. Bhandari, “Effect of honey types and concentration on starch gelatinization,”, R. J. Weston, “The contribution of catalase and other natural products to the antibacterial activity of honey: a review,”, V. Kaskoniene, P. R. Venskutonis, and V. Ceksterytė, “Carbohydrate composition and electrical conductivity of different origin honeys from Lithuania,”, E. Anklam, “A review of the analytical methods to determine the geographical and botanical origin of honey,”, S. Ouchemoukh, P. Schweitzer, M. Bachir Bey, H. Djoudad-Kadji, and H. Louaileche, “HPLC sugar profiles of Algerian honeys,”, S. Bogdanov, “Honey composition. To investigate the fact that the antibacterial activity of honey is not only due to the activity of glucose oxidase, some studies have shown that adding catalase to honey is insufficient to remove all the antibacterial activity. Book of honey,” in, S. Bogdanov, K. Ruoff, and L. Persano Oddo, “Physico-chemical methods for the characterisation of unifloral honeys: a review,”, S. Bogdanov and M. Gfeller, “Classification of honeydew and blossom honeys by discriminant analysis,”, A. Bentabol Manzanares, Z. Hernández García, B. Rodríguez Galdón, E. M. Rodríguez-Rodríguez, and C. Díaz Romero, “Physicochemical characteristics and pollen spectrum of monofloral honeys from Tenerife, Spain,”, S. Gomes, L. G. Dias, L. L. Moreira, P. Rodrigues, and L. Estevinho, “Physicochemical, microbiological and antimicrobial properties of commercial honeys from Portugal,”, P. J. Taormina, B. A. Al-Jabri, B. Nzeako, Z. Al Mahrooqi, A. Al Naqdy, and H. Nsanze, “In vitro antibacterial activity of Omani and African honey,”, A. Alnaqdy, A. Al-Jabri, Z. It is clear that this feature is due to more than one factor. Another advantage is that so consumed, honey destroys a considerable percentage of pathogenic flora in the mouth, the throat, nasal passages and the larynx. 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