This experiment compares the effect of different types of honey on bacteria growing on agar plates. Molan also highlights that dark-coloured honey obtained from the mountains of central Europe has a particularly high antibacterial activity compared to the light variant from the same region . High-performance liquid chromatography (HPLC) analysis has been used to identify the phenolic compounds in two honey extracts from north east Portugal. observed that in northern India, honey moisture content ranged between 14.63 and 21.8% . This experiment compares the effect of different types of honey on bacteria growing on agar plates. 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 . 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. This experiment compares the effect of different types of honey on bacteria growing on agar plates. Wilkinson and Cavanagh investigated the antibacterial activity of 13 honey varieties against E. coli and P. aeruginosa. Honey inhibine number and its relationship with honey concentration. 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. None of the samples was active at 1%, whereas all samples had inhibitory effects on the growth of E. coli and P. aeruginosa at 2.5% (w/v). This means that new strategies are needed to prevent and treat infectious disease. Most pathogenic bacteria failed to grow at the 40% concentration of honey and above, and the mechanism was explained through the following reasons: (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. Thats because i'm afraid the results will be too similar. This highlights the role of other important factors that can contribute to the effect of hydrogen peroxide and the acidity in the antibacterial activity of honey . A progressive increase in the inhibition was reported for most honey samples at the highest concentration in this study (at 20% at least 75% inhibition) except for K. pneumoniae which interestingly showed no inhibition at all. In an earlier report I have demonstrated the effect of direct sunlight and of the fluorescent light on the glucose oxidase activity, hence on the natural properties of honey. It is produced enzymatically in honey and can be an important feature in its antibacterial activity. We are committed to sharing findings related to COVID-19 as quickly as possible. Other phenolic compounds were present in similar quantities, but these were not specifically identified due to a lack of analytical standards . Antibiotic resistance and chronic wound infections have increased the interest in antimicrobial treatments, including honey-based wound care products, and these have been registered with medical regulatory authorities as wound care agents in many countries, among others, the European Union, USA, and New Zealand. This demonstrates the wide range of compounds that could contribute to the antibacterial properties of honey. Antibacterial effects of honey – experiment, Published 30 May 2008, Updated 28 May 2015. Divisions of Nutrition and Food Sciences, Princess Nourah Bint Abdulrahman University, Riyadh, Saudi Arabia, The osmotic effect of the honey caused shrinkage and disruption among the bacterial cells, The presence of other unidentified antibacterial substances in honey, C. M. Slover, L. H. Danziger, B. Honey is an example of a naturally available product and is the only concentrated sweetener that can be found in nature. The main enzymes in honey are invertase (saccharase) and diastase (amylase) which are introduced to honey by bees. 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. This method is usually used to establish the MIC and also MBC values in conjunction with the standard plate count. During the time in which the plate is incubating, the honey diffuses out into the agar from its point of application. 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 . For at least 2700 years, honey has been used by humans to treat a variety of ailments through topical application, but only recently have the antiseptic and antimicrobial properties of honey been discovered. 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]. 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]. Honey of different botanical origin and geographical area showed wide range of variation in their antibacterial potency. et al.rted the antibacterial properties of honey against two laboratory isolates e.g. The antimicrobial activity of this product is highly complex. Undiluted honey and its 1 : 2 to 1 : 6 aqueous dilutions showed activity of 100% and 96.4%, respectively, against P. aeruginosa and E. coli. Further methods that focus on the assessment of a growth indicator (e.g., a specific metabolite such as lactic acid), or direct microscopic counts can also be used . 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 . Herbs, plants extracts, essential oils, and honey are the most common sources for these new active compounds , and these products have been found to be effective against a range of bacterial infections and inflammatory cases . Adv Biotech & Micro 4(5): AIBM.MS.ID.555660 (2017) 0054 Abstract This study was aimed to determine the antibacterial activity of honey and/or lemon juice on strains of Streptococcus pneumoniae and Inhibine is a term that has been used to define the antibacterial agent in honey, with the “inhibine number” being used to describe the degree of dilution to which a particular type of honey keeps its antibacterial activity. Not all of the factors listed are present in all types of honey, and these compounds must be tested for and considered for clinical applications 3. The ten spices with the most potent antibacterial effects were garlic, onion, allspice, oregano, thyme, cinnamon, tarragon, cumin, cloves and lemon grass. Blossom honey should have a G + F of 60 g/100 g or higher, whereas in honeydew honey, the G + F content is much lower at 45 g/100 g with a F/G average ratio of between 1.2 and 1.3 [19, 20]. They tested the growth of bacteria in media which contained different concentrations of honey, namely, 10%, 20%, 30%, 40%, and 50% (w/v). The size of the clear zone around the honey application site, zone of inhibition (ZOI), is a measure of the potency of the honey being tested. Antibacterial activity of Manuka honey has been documented for several bacterial pathogens, however there is no information on Clostridium difficile, an important nosocomial … This was supported by another study in which solutions of pasture honey 25% (w/v) showed no detectable antibacterial activity in the presence of catalase but an activity equivalent to 14.8% phenol without catalase, whereas the same solution of Manuka honey had activity equivalent to 13.2% with and without catalase . The author declares that there are no conflicts of interest. However, gentamicin showed generally lower antibacterial activity when used in concentrations of 8.0 and 4.0 μg/ml . Different proteins have been detected in different honey varieties, predominantly related to different types of honeybees or different types of plants/flowers ; however, a group of major royal jelly proteins are shared by all honeybees. 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. 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. Antibacterial effects of honey – experiment Honey is antibacterial and can prevent growth of most types of bacteria. Myrrh extract. This clearly demonstrates that the pronounced antibacterial activity of New Zealand Manuka honey may be linked to it being rich in MGO . All honey samples as well as artificial honey were tested at a number of concentrations (1%, 2.5%, 5%, and 10% (w/v)). Although a high concentration of sugar and a low water activity will stop the growth of many microorganisms such as Staphylococcus aureus, studies have shown that often no effective bacterial inhibition occurs in the presence of “artificial” honey which can be prepared using a mixture of mono-and disaccharides at the same concentrations as those present in honey. One study compared the activity against P. aeruginosa and E. coli. Another advantage is that so consumed, honey destroys a considerable percentage of pathogenic flora in the mouth, the throat, nasal passages and the larynx. Generation of hydrogen peroxide, bee defensin-1, high osmolarity and low value of pH seems to be crucial for its antimicrobial potential. 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 . 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% . Another aspect of the studies was susceptibility of different bacteria to honey. Honey has been reported to aid in wound healing, as it has special antibacterial and antibiotic properties . A study by Alnaqdy et al. The MIC value for multifloral honey (the lowest concentration of honey, which caused visible inhibition of growth of S. aureus strain), was 3.12% in row 4 and 1.56% in rows 5 and 6. 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 . Moreover, 80% of people depend on these types of treatment in Asian countries such as China and India. MIC is used to determine the in vitro activity of an antibacterial substance and can be defined as the lowest concentration of an antibacterial agent that will inhibit the visible growth of microorganisms after an overnight incubation . Studies on the antibacterial activity of honey. The inhibine was identified as hydrogen peroxide, a main antibacterial compound in honeys . In one early study, Jeddar et al. Overall, the antibacterial activity of honey has been proven although there are contrasting results between researchers as to what concentration is effective and what is not. This is especially important since the current rise in the number of antibiotic-resistant microbial species highlights the need to source other antibacterial substances. The dark, sticky nectar is known as the “healing honey” for a reason: it has antiviral and antibacterial properties that have been used to battle bugs for centuries. 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. Furthermore, a low level of catalase would also mean a high level of hydrogen peroxide. The healing properties of honey can be attributed directly to honey bees and the enzymes the bees use to ‘process’ the honey. A linear correlation between the honey content of hydrogen peroxide and the antibacterial activity of honeys has also been reported . Although the enzyme, glucose oxidase, is naturally present in honey, it is inactive in undiluted honey because of the low pH conditions . In another study, thirty samples of honey from different parts of Oman were investigated for their activity against S. aureus. Activity against P. aeruginosa was less common in Omani honey (38%) but more common in African honey (75%) . 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. According to the results of Estevinho et al., dark honey has a high level of phenolic compounds and this has been shown to have a good correlation with its higher antibacterial activity . Interestingly, most honey samples showed no antibacterial activity in the presence of catalase except Manuka honey . It is clear that this feature is due to more than one factor. It was originally believed that hydrogen peroxide is the only factor responsible for the antibacterial effect of diluted honey, and this antibacterial activity of honey could be completely removed by the addition of catalase [50, 51]. 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). The nature of nonperoxide antibacterial activity in Manuka honey was reported by Snow and Manley-Harris using S. aureus in alkaline honey solution. It is now understood that honey is not just sugar syrup with certain physical properties that make … 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) . This study aims to compare the effects of five types of honey (both imported and local Saudi honey) against Staphylococcus aureus. The effects of flavonoids such as pinocembrin and rutin were shown to correlate with antibacterial activity of honey. Cells become dehydrated and unable to grow and proliferate in hypertonic sugar solution. It is evident that the antibacterial activity of honey can vary quite considerably and different microorganisms have different susceptibilities to different types and concentrations of honey. 12,16,17,20 In this experiment, bacterial susceptibility to honey extracts varied, which indicated a strain-dependent effect. Honey exhibits antimicrobial activities against a wide range of bacteria in different milieu. A. Ghamdi, M. J. Ansari, Y. Al-Attal, A. Al-Mubarak, and K. Salom, “Differences in composition of honey samples and their impact on the antimicrobial activities against drug multiresistant bacteria and pathogenic fungi,”, G. Brandi, P. Sestili, M. A. Pedrini, L. Salvaggio, F. Cattabeni, and O. Cantoni, “The effect of temperature or anoxia on, J. 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 . A. Imlay and S. Linn, “Bimodal pattern of killing of DNA-repair-defective or anoxically grown, K. Brudzynski, “Effect of hydrogen peroxide on antibacterial activities of Canadian honeys,”, D. Adcock, “The effect of catalase on the inhibine and peroxide values of various honeys,”, K. Brudzynski, K. Abubaker, and D. Miotto, “Unraveling a mechanism of honey antibacterial action: polyphenol/H, H. A. L. Wahdan, “Causes of the antimicrobial activity of honey,”, J. Lachman, M. Orsák, A. Hejtmánková, and E. Kovářová, “Evaluation of antioxidant activity and total phenolics of selected Czech honeys,”, J. M. Stephens, R. C. Schlothauer, B. D. Morris et al., “Phenolic compounds and methylglyoxal in some New Zealand manuka and kanuka honeys,”, I. C. F. R. Ferreira, E. Aires, J. C. M. Barreira, and L. M. Estevinho, “Antioxidant activity of Portuguese honey samples: different contributions of the entire honey and phenolic extract,”, M. Biesaga and K. Pyrzynska, “Liquid chromatography/tandem mass spectrometry studies of the phenolic compounds in honey,”, L. Yaoa, Y. Jiang, R. Singanusong, N. Datta, and K. Raymont, “Phenolic acids in Australian Melaleuca, Guioa, Lophostemon, Banksia and Helianthus honeys and their potential for floral authentication,”, R. J. Weston, L. K. Brocklebank, and Y. Lu, “Identification and quantitative levels of antibacterial components of some New Zealand Manuka honeys,”, R. J. Weston, K. R. Mitchell, and K. L. Allen, “Antibacterial phenolic components of New Zealand manuka honey,”, E. Mavric, S. Wittmann, G. Barth, and T. Henle, “Identification and quantification of methylglyoxal as the dominant antibacterial constituent of Manuka (, J. Atrott and T. Henle, “Methylglyoxal in Manuka honey—correlation with antibacterial properties,”, C. J. Adams, C. H. Boult, B. J. Deadman et al., “Isolation by HPLC and characterisation of the bioactive fraction of New Zealand manuka (, C. J. Adams, M. Manley-Harris, and P. C. Molan, “The origin of methylglyoxal in New Zealand manuka (, M. J. After analysis, the scientists concluded that the vast majority of honey's antibacterial properties come from that protein. Of these, 43% of honey samples showed excellent anti S. aureus activity. Allen et al., tested 345 samples of honey against S. aureus in the agar well diffusion assay with phenol as the reference standard. This review will focus on floral honey. 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) . The MIC … In this study, E. coli showed more susceptibility to inhibition by the honey than P. aeruginosa . Moreover, Brudzynski and Miotto reported a good correlation between honey colour, total phenolic content, levels of Maillard reaction-like products (MRLPs), antioxidant activity, and the antibacterial activity of unheated honey . 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 . Sign up here as a reviewer to help fast-track new submissions. 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. 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 TP contents ranged between 21.3 and 184.3 mg/kg and the main phenolic acid in all honey samples was gallic acid with 4.52, 4.11, 1.39, and 3.63 mg/100 g, respectively, for the different honey types mentioned above . This suggests that honey contains other important components with antibacterial properties. The researcher identifies the antibacterial properties of honey with isolated compounds derived from honey, selected antibiotics. Other important components of honey are the enzymes present which contribute to its antioxidant and antibacterial activities. Propolis, a flavonoid-rich product of honey comb, exhibits antibacterial and anti-inflammatory properties (Bosio et al., 2000) which is very powerful *Corresponding author. 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 . They found that 81% of the Omani honey samples and 88% of the African honey samples assessed in the study had antistaphylococcal activity, but only 63% of Omani honeys and 62% of African honeys showed any activity against E. coli. Basson and Grobler tested the antibacterial potency of different honey varieties produced from indigenous wild flowers grown in South Africa against S. aureus. 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. Infected mice have been used to study the effect of honey on wound infection. The most potent honeys, such as Manuka, dark buckwheat, Heather, or chestnut honeys, have their MIC values, ranging from 1% to 12.5% (w/v). 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 . This discovery has provoked an increase in the number of studies that have investigated the effect of substances other than peroxide activity. studied the antibacterial activity of 24 samples of honey (16 from Oman and eight from Africa) against three bacteria, namely, S. aureus, E. coli, and P. aeruginosa. Each tube or well is inoculated with the standardized test microorganisms and incubated. including antibacterial, antifungal, antidiabetic, anticancer and antiviral activities . Some of the differences in the composition of honey are due to the differences between regions (floral sources) but seasonal differences can also be important . 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 . 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. Protein content in honey is very low and ranges between 0.1 and 0.5%. In contrast, S. aureus has been found to be completely inhibited by one honey variety at 17% when impregnated in nutrient agar . 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 . 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. There are several other methods that have been used to measure the antibacterial activity of honey. 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,”. Monosaccharides fructose and glucose125 in honeys [ 30 ] the current rise in the from... Invertase ( α-glucosidase ), and storage is the most appropriate method concentrations of 8.0 and 4.0 μg/ml [ ]... 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