In addition, studying the effect of honey on the growth of bacteria such as S. aureus, which has a high tolerance of low water activity, gives clear evidence that the antibacterial activity of honey must also be attributed to other factors. evaluated the antibacterial effect of pure honey in vitro. I'm testing how effective different types of honey is against e coli k12, probloem is i need a way of measuring the antibacterial properties of this honey that doesn't involve measuring the range of inhibition. This means that new strategies are needed to prevent and treat infectious disease. 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. Honey prevents microbial growth through the use of hydrogen peroxide (H2O2), methylglyoxal (MGO), bee defensin-1, flavonoids, and a relatively low pH (~3.3) 13. Crystallized honey is quite suitable for that purpose. et al.rted the antibacterial properties of honey against two laboratory isolates e.g. Bacterial susceptibility to honey can be measured quantitatively by several methods, broth (micro) dilution assay, well/disk diffusion assay, agar dilution methods, and time-kill assay. The results showed the presence of 14 phenolic compounds which were mainly phenolic acids and flavonoids. Honey is an example of a naturally available product and is the only concentrated sweetener that can be found in nature. More recently, honey has been reported to have an inhibitory effect on approximately 60 species of bacteria, including aerobes and anaerobes, gram-positive and gram-negative bacteria [ 5 ]. It is evident that undiluted honey has the ability to stop the growth of bacteria completely because of the high content of sugar; high sugar concentration of honey exerts osmotic pressure on bacterial cells which causes transport of water out of bacterial cells through osmosis. antibacterial applicant (Miorin et al., 2003). As a result, several ancient methods have been re-evaluated and the substances/procedures employed historically to cure diseases are now attracting renewed scientific attention. Also, honey was active against biofilms formed by methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant Staphylococcus aureus (MRSA), and Pseudomonas aeruginosa with bactericidal rates ranging from 63–82%, 73–63%, and 91–91%, respectively, that was higher than the effect of commonly used single antibiotics commonly used [78]. This discovery has provoked an increase in the number of studies that have investigated the effect of substances other than peroxide activity. A study of the biological activity of chestnut, Herero floral, and Rhododendron honeys obtained from Anatolia in Turkey revealed activity against all the test microorganisms but the extracts gave rise to moderate inhibition against only a few microorganisms, e.g., H. pylori and S. aureus [38]. Nanda et al. S. aureus needs an aw of lower than 0.86 for complete inhibition which is equivalent to a concentration of honey of 29% (v/v) [10]. Moreover, the world today needs further assessments of natural substances that can be used to combat microorganisms with minimal side effects or consequences of overdose or high consumption. More research is needed in this area. A. Niemira, and L. R. Beuchat, “Inhibitory activity of honey against foodborne pathogens as influenced by the presence of hydrogen peroxide and level of antioxidant power,”, J. Bertoncelj, U. Dobersek, M. Jamnik, and T. Golob, “Evaluation of the phenolic content, antioxidant activity and colour of Slovenian honey,”, L. Estevinho, A. P. Pereira, L. Moreira, L. G. Dias, and E. Pereira, “Antioxidant and antimicrobial effects of phenolic compounds extracts of Northeast Portugal honey,”, K. L. Allen, P. C. Molan, and G. M. Reid, “A survey of the antibacterial activity of some New Zealand honeys,”, V. Nanda, B. C. Sarkar, H. K. Sharma, and A. S. Bawa, “Physico-chemical properties and estimation of mineral content in honey produced from different plants in Northern India,”, T. Marshall and K. M. Williams, “Electrophoresis of honey: characterization of trace proteins from a complex biological matrix by silver staining,”, S. Serrano, R. Espejo, M. Villarejo, and M. L. Jodral, “Diastase and invertase activities in Andalusian honeys,”, P. Vit and P. Pulcini, “Diastase and invertase activities in Meliponini and Trigonini honeys from Venezuela,”, J. W. White Jr., M. H. Subers, and A. I. Schepartz, “The identification of inhibine, the antibacterial factor in honey, as hydrogen peroxide and its origin in a honey glucose-oxidase system,”, J. M. Andrews, “Determination of minimum inhibitory concentrations,”, T. Patton, J. Barrett, J. Brennan, and N. Moran, “Use of a spectrophotometric bioassay for determination of microbial sensitivity to manuka honey,”, S. Bogdanov, “Nature and origin of the antibacterial substances in honey,”, P. E. Lusby, A. L. Coombes, and J. M. Wilkinson, “Bactericidal activity of different honeys against pathogenic bacteria,”, D. J. Willix, P. C. Molan, and C. G. Harfoot, “A comparison of the sensitivity of wound-infecting species of bacteria to the antibacterial activity of manuka honey and other honey,”, R. A. Cooper, P. C. Molan, and K. G. Harding, “Antibacterial activity of honey against strains of, M. S. Osato, S. G. Reddy, and D. Y. Graham, “Osmotic effect of honey on growth and viability of, M. Küçük, S. Kolaylı, Ş. Karaoğlu, E. Ulusoy, C. Baltacı, and F. Candan, “Biological activities and chemical composition of three honeys of different types from Anatolia,”, M. M. Cavia, M. A. Fernández-Muiño, S. R. Alonso-Torre, J. F. Huidobro, and M. T. Sancho, “Evolution of acidity of honeys from continental climates: influence of induced granulation,”, M. Ali, “Hydrogen peroxide therapies: recent insights into oxystatic and antimicrobial actions,”, L. M. Bang, C. Buntting, and P. Molan, “The effect of dilution on the rate of hydrogen peroxide production in honey and its implications for wound healing,”, S. Bogdanov, “Characterisation of antibacterial substances in honey,”, L. A. Roth, S. Kwan, and P. Sporns, “Use of a disc-assay system to detect oxytetracycline residues in honey,”, K. Brudzynski, K. Abubaker, L. Martin, and A. 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. All samples were tested at different concentrations (0.1%, 1%, 5%, 10%, and 20% (w/v)). To illustrate this, Lachman et al., evaluated the total polyphenol content of honey varieties harvested in the period from May to August 2006 and found the highest TP acid content occurred in the honey collected at the beginning of June (on average 170.21 mg/Kg) and July (on average 163.32 mg/Kg), whereas it was much lower in samples (83.60 mg/Kg) collected during the other months [53]. A good linear correlation has been shown to exist between MGO content and the antibacterial activity of Manuka honey [61]. The antibacterial effect of honey refers to the experiment that identify the effect of honey on different bacteria growing on agar plates in order to find out the properties present in the honey that help to destroy the pathogenic bacteria as measured by Kirby Bauer method. Castle, “Re-examining the role of hydrogen peroxide in bacteriostatic and bactericidal activities of honey,”, K. Brudzynski and R. Lannigan, “Mechanism of honey bacteriostatic action against MRSA and VRE involves hydroxyl radicals generated from honey’s hydrogen peroxide,”, N. Al-Waili, A. In another study, thirty samples of honey from different parts of Oman were investigated for their activity against S. aureus. 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. Although it is believed that some of these enzymes come from nectar, [28] it is known that the α-amylase and α-glucosidase in honey comes from bee salivary secretions [29]. 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 bacterial growth (change in turbidity) is assessed spectrophotometrically. Cells become dehydrated and unable to grow and proliferate in hypertonic sugar solution. High-performance liquid chromatography (HPLC) analysis has been used to identify the phenolic compounds in two honey extracts from north east Portugal. This matter has been investigated by WHITE and myself. antibacterial effect of honey during its extraction, processing, and storage is the direct light. The antibacterial effects of honey have been known in practical terms for over a hundred years in the absence of a proper understanding of their specific mechanisms of action. This is similar to the findings of other studies and demonstrates the consistency amongst different varieties in terms of the key components [15, 16]. In general, honeys might be classified to two groups: honeys whose activity is hydrogen-peroxide dependent (honeys of American, European, and some Asian origin) and honeys whose activity depends on the presence of methylglyoxal, like New Zealand Manuka honey. Honey has been shown to have a strong activity against many bacteria in both media and in culture. 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. A similar range of hydrogen peroxide concentrations (1 mM to 2.5 mM) was enough to kill E. coli in 15 minutes [47, 48]. Indeed, the maximum level of hydrogen peroxide produced can be obtained from a 30–50% honey dilution [10], potentially ranging between 5 and 100 µg H2O2/g honey (which is equivalent to ∼0.146–2.93 mM) [30]. Of these, 43% of honey samples showed excellent anti S. aureus activity. Protein content in honey is very low and ranges between 0.1 and 0.5%. Other dark-coloured honeys have also demonstrated high antibacterial activity such as sweet chestnut honey (Castanea sativa), Manuka honey (Leptospermum scoparium), and Heather honey (Calluna vulgaris) [25]. Some components of these raw materials possess important antibacterial properties that can contribute to the total antibacterial activity of honey [12]. in 2005 which characterised the effect of honey on the adherence of Salmonella to intestinal epithelial cells showed that a honey dilution of 1 : 8 reduced the adherence from 25.6 ± 6.5 to 6.7 ± 3.3 bacteria per epithelial cell [73]. The increase in the resistance of pathogenic bacteria to antibiotics is also an increasingly important factor behind the growing interest in the use of these natural compounds. Scientists have found that natural materials are generally more acceptable to consumers, and if these alternative approaches are effective, this may reduce the reliance on more synthetic substances [2]. These enzymes have been studied and have been shown to contain similar properties of antibiotics. Interestingly, most honey samples showed no antibacterial activity in the presence of catalase except Manuka honey [25]. According to the World Health Organization (WHO) statistics, up to 80% of the population in some developed countries have used natural products in their primary health care [1]. It is important to note that the level of hydrogen peroxide in honey is also determined by the presence and action of catalase. The increase in their popularity is due to their potent activities and generally very low toxicity. This indicates that obviously much lower than the 29% honey that would be required if the effect was based solely on water activity [35, 36]. This review covers the antibacterial activity of honey, its use in the treatment of infection and diseases, and the features that are relevant to its activity. Manuka honey is considered to have a unique factor (unique Manuka factor (UMF)) responsible for its antibacterial activity, and this is considered to be MGO. The result showed that the South African honey varieties did not have strong bactericidal activity, and honey concentration above 25% was necessary for antibacterial activity, due to the osmolality and carbohydrate concentration [67]. Hydrogen peroxide (H2O2) is an important oxidizing and sanitizing agent [40]. The antibacterial activity ranged between 2% to 58% (w/v) with a median of 13.6%. High amounts of MGO are found in Manuka honey, up to around 800 mg/kg (up to 100-fold) higher compared to conventional honey [60, 62, 63]. Most pathogenic bacteria failed to grow at the 40% concentration of honey and above, and the mechanism was explained through the following reasons: [71](1)The osmotic effect of the honey caused shrinkage and disruption among the bacterial cells(2)The low pH(3)The presence of other unidentified antibacterial substances in honey. They tested the growth of bacteria in media which contained different concentrations of honey, namely, 10%, 20%, 30%, 40%, and 50% (w/v). 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]. The most commonly used bacterial susceptibility assay is a broth micro- or macrodilution assay. During the time in which the plate is incubating, the honey diffuses out into the agar from its point of application. 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). A. Mahrooqi, B. Nzeako, and H. Nsanze, “Inhibition effect of honey on the adherence of Salmonella to intestinal epithelial cells in vitro,”, N. S. Al-Waili, “Investigating the antimicrobial activity of natural honey and its effects on the pathogenic bacterial infections of surgical wounds and conjunctiva,”, T. R. Shamala, Y. P. Shri Jyothi, and P. Saibaba, “Antibacterial effect of honey on the in vitro and in vivo growth of, J. M. Wilkinson and H. M. A. Cavanagh, “Antibacterial activity of 13 honeys against, C. Badet and F. Quero, “The in vitro effect of manuka honeys on growth and adherence of oral bacteria,”, T. Alandejani, J. Marsan, W. Ferris, R. Slinger, and F. Chan, “Effectiveness of honey on, O. E. Adeleke, J. O. Olaitan, and E. I. Okpekpe, “Comparative antibacterial activity of honey and gentamicin against, E. O. Agbaje, T. Ogunsanya, and O. I. Aiwerioba, “Conventional use of honey as antibacterial agent,”. Furthermore, the concentration of MGO increases as Manuka honey matures and after storage (up to 120 days) at 37°C, which has been attributed to the nonenzymatic conversion of dihydroxyacetone to MGO during long-term storage [62]. Thats because i'm afraid the results will be too similar. 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]. Methyl syringate (MSYR) was the major product in phenolic extracts of active Manuka honey isolated by Weston et al., comprising more than 45% of the TP [59]. Nevertheless, the content of individual carbohydrates did vary and ranged between 329.2 to 426.3 mg/g for fructose and glucose (as the dominant components) [13]. This experiment compares the effect of different types of honey on bacteria growing on agar plates. Honey contains antioxidants and flavonoids that may function as antibacterial agents. 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. All honey samples as well as artificial honey were tested at a number of concentrations (1%, 2.5%, 5%, and 10% (w/v)). The effects of flavonoids such as pinocembrin and rutin were shown to correlate with antibacterial activity of honey. Antibacterial effects of honey – experiment Honey is antibacterial and can prevent growth of most types of bacteria. As a result, many studies have analysed the composition of honey and have studied the physical and chemical properties that may give rise to its ability to work against various microorganisms [7]. These include glucose oxidase, invertase (α-glucosidase), catalase, diastase (α-and β-amylase), and peroxidase. 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]. Activity against P. aeruginosa was less common in Omani honey (38%) but more common in African honey (75%) [72]. These products are mainly based on Manuka (Leptospermum … This experiment compares the effect of different types of honey on bacteria growing on agar plates. Antibacterial activity of honey is mainly dependent on a combination of its peroxide activity and non-peroxide components. Honey has drawn increasing attention as a remedy for wound treatment of different kinds, mainly due to a verified antibacterial activity [1]. Manuka honey, meanwhile, has a phenolic acid content that ranges between 430–2706 mg/kg compared with Kanuka honey (424–1575 mg/kg) collected at the same time and from the same site [54]. A study by Alnaqdy et al. The inhibine was identified as hydrogen peroxide, a main antibacterial compound in honeys [30]. Honey is one such product that used to be widely used to combat bacteria. This review will focus on floral honey. No inhibition was observed at 0.1% but the 1% concentration showed some inhibition with C. freundii, E. coli, M. phlei, and three species of Salmonella. 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]. The first explanation of the antibacterial activity of honey was reported in 1892 by Van Ketel [10]. The samples included Kanuka, Manuka, Heather, and Kamah honey. Despite the developments in controlling infectious disease around the world, they are still the second biggest cause of morbidity and mortality due in part to the increase in drug resistance among large numbers of the bacterial strains. Thirty-eight percent of S. aureus strains were killed by 50% honey in 30 minutes and 45% after one hour. 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. The antibacterial properties of honey should be examined on non-pathogenic bacteria, such as E. … 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. Bogdanov studied the antibacterial activity of eleven types of honey, including the common varieties such as acacia, blossom, chestnut, lavender, and orange against Staphylococcus aureus and Micrococcus luteus and found that the inhibition of the different honey varieties ranged from 37 to 74% [33]. According to some studies, honey has been shown to possess a high level of phenolic compounds which might contribute to its antibacterial activity. In addition, the mild acidity and hydrogen peroxide content have obvious antimicrobial effects [34]. Allen et al., tested 345 samples of honey against S. aureus in the agar well diffusion assay with phenol as the reference standard. coli However, the sensitivity of bacteria to hydrogen peroxide produced in honey can be influenced by the presence of phytochemical compounds in honey [44]. Many of these substances have been discovered to have similar inhibitory effect and mechanisms of action to antibiotics, causing damage to bacterial cell walls as well as affecting protein synthesis in bacterial cells [5]. Thus, part of the antimicrobial honey components are absorbed while still in the mouth which makes the consumption much more complete. Nevertheless, some bacterial strains are more sensitive to the osmotic effects of carbohydrate monomers and dimers than others, and it has been shown that a concentration of 15% (w/v) carbohydrate (fructose, glucose, and glucose and fructose combinations) was sufficient to have a similar inhibitory effect as honey on all 28 tested isolates of Helicobacter pylori [37]. In general, it is important to appreciate that the results will depend largely on the technique and scientific judgment, and this needs to be considered when comparing results using different methods [32]. Studies have shown the antibacterial activity of catalase-treated honey, the nonperoxide antibacterial activity (NPABA), has been identified. This antibacterial action will be reduced when honey is diluted by body fluids at the site of infection. The agar diffusion assay technique, for example, is a method in which a small quantity of honey or solution of honey is applied to the centre of a well (about 6 mm in diameter) cut into nutrient agar plate previously inoculated with a microbial culture [10]. The colour of honey ranges from light yellow, through to amber and dark reddish amber to a nearly black colour [23]. The susceptibility of Campylobacter jejuni to the antibacterial activity of Manuka honey was also tested, and the results showed that 1% (v/v) of Manuka honey was sufficient to give the minimum inhibitory effect [69]. Measurement of absorbance using fluorimetry or the spectrophotometric determination of growth has a greater sensitivity especially when used with low honey concentrations [32]. 2019, Article ID 2464507, 10 pages, 2019. https://doi.org/10.1155/2019/2464507, 1Divisions of Nutrition and Food Sciences, Princess Nourah Bint Abdulrahman University, Riyadh, Saudi Arabia. The effect of a 10-fold excess catalase upon the antibacterial assay was examined but no statistical difference was evident in the outcome between the normal amount of catalase and the 10-fold excess, thus indicating that nonperoxide antibacterial activity was not due to residual hydrogen peroxide [64]. Honey has been in use as a wound dressing for thousands of years.1,2 In the past few decades, there has been a large amount of clinical evidence has been accumulated that demonstrates the effectiveness of honey in this application.3,4 However, it is only in more recent times that the science behind the efficacy has become available. Of the solids in honey, 84% is a mixture of the monosaccharides fructose and glucose125. 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]. 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