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Hydrogen Peroxide Concentrations in Different Honey Solutions After 24-h Incubation at 37°C

Hydrogen Peroxide Concentrations in Different Honey Solutions After 24-h Incubation at 37°C

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Abstract Although hydrogen peroxide (H2O2) is one of the major antibacterial factors in most honeys, it does not accumulate in medical-grade manuka honey. The goal of this study was to investigate the effect of artificially added methylglyoxal (MGO) on H2O2 accumulation in natural non-manuka honeys. H2O2 concentrations in the honey solutions were d...

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... most potent producers were the two honeydew honeys, whose 50% solutions accumulated 306.9 -6.8 and 495.8 -9.1 lM H 2 O 2 , respectively. Significantly lower lev- els of H 2 O 2 were found in manuka and acacia honey (Table 1). In our very recent study, we documented that high concentrations of MGO present in manuka honeys modify defensin 1, an antibacterial bee-derived peptide and also the dominant protein in honey-MRJP1. ...

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... Since few studies have analysed large numbers of manuka/Leptospermum honeys for physicochemical properties such as pH, sugar content, colour, total phenolics content and antioxidant activity, comparison of the current data to previous studies is limited, however, the available data are broadly similar [9,32,33]. Manuka honeys produced very low levels of hydrogen peroxide, which was expected due to the known interference of MGO with the glucose oxidase enzyme [34]. ...
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Variation in the antibacterial potency of manuka honey has been reported in several published studies. However, many of these studies examine only a few honey samples, or test activity against only a few bacterial isolates. To address this deficit, a collection of 29 manuka/ Leptospermum honeys was obtained, comprising commercial manuka honeys from Australia and New Zealand and several Western Australian Leptospermum honeys obtained directly from beekeepers. The antibacterial activity of honeys was quantified using several methods, including the broth microdilution method to determine minimum inhibitory concentrations (MICs) against four species of test bacteria, the phenol equivalence method, determination of antibacterial activity values from optical density, and time kill assays. Several physicochemical parameters or components were also quantified, including methylglyoxal (MGO), dihydroxyacetone (DHA), hydroxymethylfurfural (HMF) and total phenolics content as well as pH, colour and refractive index. Total antioxidant activity was also determined using the DPPH* (2,2-diphenyl-1-picrylhydrazyl) and FRAP (ferric reducing–antioxidant power) assays. Levels of MGO quantified in each honey were compared to the levels stated on the product labels, which revealed mostly minor differences. Antibacterial activity studies showed that MICs varied between different honey samples and between bacterial species. Correlation of the MGO content of honey with antibacterial activity showed differing relationships for each test organism, with Pseudomonas aeruginosa showing no relationship, Staphylococcus aureus showing a moderate relationship and both Enterococcus faecalis and Escherichia coli showing strong positive correlations. The association between MGO content and antibacterial activity was further investigated by adding known concentrations of MGO to a multifloral honey and quantifying activity, and by also conducting checkerboard assays. These investigations showed that interactions were largely additive in nature, and that synergistic interactions between MGO and the honey matrix did not occur.
... The dihydroxyacetone is converted non-enzymatically by the Maillard reaction into MGO (Nolan et al., 2019). It is believed that the presence of MGO in the honey inhibits the glucose peroxidase enzyme as no hydrogen peroxide is produced (Majtan et al., 2013). Both honey types contain additional molecules that exert direct antimicrobial effects, including polyphenolic compounds (phenolic acids, flavonoids, and tannins) and antimicrobial peptides such as bee defensin-1. ...
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Hospital-acquired infections and treatment-related wound complications constitute a tremendous burden for the health care system, particularly given the serious increase in multidrug resistant pathogens. Imagine that a large part of nosocomial infections can be prevented using a simple treatment. In this respect, honey is used mainly in topical cutaneous wound care because of its potent broad-spectrum antibacterial and wound healing activities. However, therapeutic use outside this scope has been limited. The current review provides an in-depth view of studies using honey outside the conventional wound care indications. Non-conventional routes of honey application include subcutaneous, intra-socket, abdominal, and oral administration in novel indications, such as post colon surgery, mucositis, and tooth extraction. Honey consistently demonstrates beneficial therapeutic activities in these novel applications, orchestrating antimicrobial and prophylactic activity, reducing inflammation and wound dehiscence, and inducing healing, epithelialization, and analgesic activity. Several molecular mechanisms are responsible for these beneficial clinical effects of honey during the course of wound healing. Pro-inflammatory effects of honey, such as induction of iNOS, IL-1β, and COX-2, are mediated by TLR4 signaling. In contrast, honey's anti-inflammatory actions and flavonoids induce anti-inflammatory and antioxidant pathways by inducing NRF2 target genes, including HO-1 and PRDX1. The molecular and biochemical pathways activated by honey during the different phases of wound healing are also discussed in more detail in this review. Variation between different honey origins exists, and therefore standardized medical-grade honey may offer an optimized and safe treatment. Honey is a valuable alternative to conventional antimicrobial and anti-inflammatory therapies that can strongly reduce nosocomial infections.
... MGO presumably acts through irreversible crosslinking of bacterial macromolecules (proteins and DNA), leading to loss of function (Brudzynski, 2021;Johnston et al., 2018;Majtan, Bohova, Prochazka, & Klaudiny, 2014;Majtan et al., 2012). ...
... The effect was observed for polyfloral and buckwheat honeys collected in Poland but also for manuka honey with high concentration of MGO (at least 500 mg/kg). In the study of Majtan, Bohova, Prochazka, and Klaudiny (2014) at concentration of 50% (v/v) none of the five tested honeys (including manuka honey) caused statistically significant detachment of Enterobacter cloacae biofilm biomass after 24 h of treatment. Moreover, both investigated acacia honeys promoted production of biofilm biomass of this pathogenic bacteria. ...
... We strongly suspect that enzymatic activity of diastase in manuka honey is affected by the presence of highly reactive MGO. Similarly, enzymatic activity of GOX in non-manuka honey spiked with MGO significantly decreased and the enzyme was structurally modified (Majtan, Bohova, Prochazka, & Klaudiny, 2014). Therefore, there is an urgent need to establish and certify more effective honey quality control methods or regularly revise current qualitative tools in order to partially address the issues of honey adulteration production and/or to identify the exceptions based on state-of-the-art knowledge of individual honey types. ...
Article
Background Honey is considered as a functional food with health-promoting properties. Its potent antibacterial and antibiofilm effects are the major attributes of so called ‘medical-grade honey’ which is topically used for the treatment of burns, wounds and skin disorders. Nevertheless, the current set of honey quality parameters adopted in the European Union do not include its biological properties. Furthermore, in light of the accelerated growth of scientific evidence, there is an urgent need to revise current qualitative tools, and to establish and certify more effective honey quality control. Scope and approach This up-to-date narrative review aims to discuss the recent clinical evidence describing the use of honey in the management of various disorders including respiratory tract infections, metabolic and gastro-intestinal derangements. Current knowledge about the antibacterial activity of honey, as the most studied biological properties of natural honey, focusing on mechanism of action and the factors/compounds responsible for the antibacterial effects is also discussed. In addition, the weaknesses of current honey quality parameters are highlighted and a new potentially qualitative parameter that takes into account honey functionality is presented. Key findings and conclusions Data from in vitro and in vivo experiments, as well as human clinical studies clearly indicate the importance and efficacy of honey as an antibacterial agent. Antibacterial activity can vary from honey to honey but must not be identical to the activity of the honey sugar content. In most cases, antibacterial activity can be negatively impacted by thermal processing and long-term storage and this activity is therefore a suitable and sensitive quality parameter. From a clinical point of view, we strongly advocate to solely use natural honey that has undergone only minimal processing in order to preserve the full spectrum of biological activities.
... A 100-fold difference in antimicrobial activity between honey types is reported (Mandal & Mandal, 2011). Although manuka honey may be extensively investigated, other types of honey have similar or stronger antimicrobial activity and may be more effective for wound healing Grego et al., 2016;Lusby, Coombes & Wilkinson, 2005;Majtan, Bohova, Prochazka & Klaudiny, 2013;Pleeging et al., 2020;Sherlock et al., 2010;Smaropoulos & Cremers, 2021). In a direct in vitro comparison study, L-Mesitran Soft had a stronger antimicrobial activity than manuka honey-based Medihoney against Staphylococci and Pseudomonas spp. ...
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South Africa is home to some of the world's most endangered wildlife, with the White rhinoceros (Cerathotherium simum) among its most threatened species due to its highly sought-after horn. Since the reproduction number is below the mortality rate, there is a decrease in rhinos year over year, urgently requiring changes and saving the survivors. In this study, the efficacy of medical grade honey (MGH) for wound care in rhinos was investigated. We presented a case series of seven rhinos with wounds of different etiologies, including gunshots and poaching. Four wounds were around the horns and three on the limbs. It was a challenge to take care and follow-up the wounds of these wild animals in their natural habitat. A well-balanced decision between the need of treatment and risk of immobilization is constantly evaluated. In the presented cases, MGH proved a novel tool with minimal intervention and maximal effectiveness. With the severity of the wounds taken into consideration, there was a rapid healing in all cases, while infection resolved when present. MGH creates a moist and anti-inflammatory wound environment, while promoting almost all aspects in the wound healing processes, such as autolytic debridement, angiogenesis and re-epithelialization. The efficacy of MGH has constantly been confirmed in other cases and literature as well. MGH forms a potent therapy for treating wounded rhinos, independent of the severity of the wound. The strong antimicrobial and healing properties make it an easy and versatile product that can be used in all kinds of wounds.
... This is an important finding, because manuka honey is considered by most researchers as the "front-runner of honeys for non peroxide antimicrobial activity" [65]. Hydrogen peroxide is not accumulated in manuka honey due to the destruction of glucose oxidase by methylglycoxal [66] and this is perhaps one reason for the outperformance of antibacterial activity of the Greek honeys. These honeys can act as both bacteriostatic and bactericidal depending on the concentration used. ...
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To study the antibacterial effect of different Greek honeys, samples of citrus honey, Saturja spp. Honey, and oregano and sage honey were collected directly from producers. Manuka honey and artificial honey were used as controls. The honeys were diluted in various concentrations to determine the minimum inhibitory concentration (MIC) and were also placed in agar wells to determine the inhibitory zones of growth. The bacteria tested were two reference strains and five pathogens isolated from patients with various dental ailments. A series of samples were diluted with artificial saliva instead of distilled water to simulate the conditions in the oral cavity. The results show that in most cases the Greek honeys, and particularly the citrus honey and the oregano and sage honey, outperformed the antibacterial activity of manuka honey against all tested bacteria. This performance was due to the hydrogen peroxide as well as to other components of the honeys, that is, peptides and other substances such as phenolic compounds and flavonoids. Artificial saliva enhanced the antibacterial effect of the honeys in comparison to distilled water.
... The Manuka honey antibacterial effect seems to be incompatible with peroxidemediated activity, as MGO supplementation has been shown to impede GOx activity in non-Manuka honeys by morphological damage and the formation of a glycated derivative structure. As a result, GOx-modulated H 2 O 2 production was significantly compromised in a dose-proportional fashion [94]. However, an interaction between peroxide and nonperoxide systems has been described by Henatsch et al., who studied the antibacterial activity of honey in relation not only to its content of MGO and analogous α-dicarbonyl compounds but also to their conversion into different free radicals in the presence of either hydrogen peroxide or amino acids (specifically arginine and lysine) [95]. ...
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Honey is a honey-bee product obtained mainly by the enzymatic processing of nectar from a variety of plants, which leads to the wide range of colours and flavours available on the market. These organoleptic and nutritional features are influenced by the chemical composition, which in turn depends on the botanical origin. Bioactive compounds account for honey beneficial activity in medical applications, which explains the extensive use of honey in ethno-pharmacology since antiquity, from cough remedies to dermatological treatments. Wound healing is one of the main therapeutic uses of honey, and various design options in pharmaceutical technology such as smart delivery systems and advanced dressings are currently being developed to potentiate honey’s valuable properties for better performance and improved final outcome. In this review, we will focus on the latest research that discloses crucial factors in determining what properties are most beneficial when considering honey as a medicinal product. We will present the most recent updates on the possible mechanisms responsible for the exceptional effects of this ageless therapeutical remedy on skin repair. Furthermore, the state-of-the-art in application techniques (incorporation into scaffolds as an alternative to direct administration) used to enhance honey-mediated wound-healing properties are explored.
... For instance, by exposing to heat, the GOx could be inactivated, but also, a catalase enzyme present in the pollen could deteriorate the H 2 O 2 . Additionally, as it was expected, the level of GOx influences the H 2 O 2 [59]. Another study also highlighted the strong connection between the presence of H 2 O 2 and the strong antibacterial activity of honey, evidencing at the same time that there are some exceptions that exhibit strong bactericidal activity but have low levels of H 2 O 2 [60]. ...
... However, even if H 2 O 2 is a major player involved in honey's bactericidal activity, upon dilution, Manuka honey does not display a detectable concentration of this element, but exhibits great antibacterial activity [59]. Nevertheless, Manuka honey contains up to 800 mg/kg of methylglyoxal (MGO) [50]. ...
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Since ancient times, honey has been considered one of the most illustrious and esteemed natural products. Honey plays two key roles; specifically, it is an appreciated nutritional product, and also exhibits a wide range of beneficial properties for human health as a therapeutic agent. Furthermore, it has been shown that honey has valuable effects on the biological and physiological features of mulberry silkworms (Bombyx mori). Bombyx mori exhibits importance not only for the economy, but it also serves as an important biotechnological bioreactor for the production of recombinant proteins that have a great impact in the medical field and beyond. It also represents an important model organism for life sciences. In view of the fact that silk fibroin serves as a natural biopolymer that displays high biocompatibility with human organisms and due to honey’s various and remarkable properties for human health, the two elements are currently used together in order to develop ideal biomaterials for a wide range of purposes. In this review, by discussing the applicability of honey on Bombyx mori and beyond, the importance of honey for life sciences and related fields is spotlighted.
... Manuka honey is probably the most often used for wound care because this was the first honey extensively investigated regarding the antimicrobial properties [24,89]. Furthermore, other types of honey have similar or superior antimicrobial activity and may be more effective for wound healing [22,[90][91][92][93][94]. Current findings are in accordance with previous in vitro results that directly compared the antimicrobial activity of L-Mesitran Soft and Medihoney against eleven Staphylococci and eleven Pseudomonas spp. ...
... The main difference between Manuka and other types of MGH is their antimicrobial mechanism. Manuka-honey-mediated antimicrobial activity is mainly attributed to methylglyoxal whereas other honey types are mainly due to hydrogen peroxide [94][95][96]. ...
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Biofilms hinder wound healing. Medical-grade honey (MGH) is a promising therapy because of its broad-spectrum antimicrobial activity and the lack of risk for resistance. This study investigated the inhibitory and eradicative activity against multidrug-resistant Pseudomonas aeruginosa biofilms by different established MGH-based wound care formulations. Six different natural wound care products (Medihoney, Revamil, Mebo, Melladerm, L-Mesitran Ointment, and L-Mesitran Soft) were tested in vitro. Most of them contain MGH only, whereas some were supplemented. L-Mesitran Soft demonstrated the most potent antimicrobial activity (6.08-log inhibition and 3.18-log eradication). Other formulations ranged between 0.89-log and 4.80-log inhibition and 0.65-log and 1.66-log eradication. Therefore, the contribution of different ingredients of L-Mesitran Soft was investigated in more detail. The activity of the same batch of raw MGH (1.38-log inhibition and 2.35-log eradication), vitamins C and E (0.95-log inhibition and 0.94-log eradication), and all ingredients except MGH (1.69-log inhibition and 0.75-log eradication) clearly support a synergistic activity of components within the L-Mesitran Soft formulation. Several presented clinical cases illustrate its clinical antimicrobial efficacy against Pseudomonas aeruginosa biofilms. In conclusion, MGH is a potent treatment for Pseudomonas biofilms. L-Mesitran Soft has the strongest antimicrobial activity, which is likely due to the synergistic activity mediated by its supplements.
... Several research groups have attempted to verify the antimicrobial effect of Manuka honey and propolis [17,[33][34][35]. Maddocks S. et al. [36] proved that Manuka honey inhibited S. aureus, P. aeruginosa, and S. pyogenes bacterial cells in binding to the human proteins, i.e., fibronectin, fibrinogen, and collagen. ...
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Chronic wound infections are difficult to manage because of the biofilm formation in the wound environment. New measures for eliminating infections are necessary to increase the chance of wound healing. Apitherapy may be the new solution. The aim of this study was to assess the prevalence of wound infection factors and to examine the impact of Manuka honey and ethanol extract of propolis on biofilm formation of Proteus mirabilis isolated from chronic wound infections. According to the findings, the most frequent factors of infection are Staphylococcus aureus (46.1%), Pseudomonas aeruginosa (35.0%), and Proteus mirabilis (10.6%). Minimal inhibitory concentration and minimal bactericidal concentration values were assigned using the microbroth dilution test according to the Clinical and Laboratory Standards Institute. Biofilm of Proteus mirabilis isolates was formed in 96-well polystyrene plates and treated with Manuka honey (concentrations from 1.88% to 30.0%) and ethanol extract of propolis (1.0% to 40.0%). After 24 h, the biofilm viability was expressed by formazan absorbance (λ = 470 nm). Manuka honey reduced the biofilm viability in all, and ethanol extract of propolis in most, of the concentrations tested. Ethanol extract of propolis at the concentrations of 20.0% and 40.0%, reduced biofilm viability stronger than ethanol itself. With these results comes the conclusion that these substances can reduce biofilm formation.
... The well-known factors that affect the biochemical composition of bee's honey are the botanical origin, geographical area, climate and storage conditions, handling, effect of biochemical constituents on each other and bee species, health and nutrition (El Sohaimy et al., 2015;Burns et al., 2018;Majtan et al., 2014). ...
... This study proved that honey samples with high diacrbonyl compounds are with low hydrogen peroxide percentage similar to the nding of Majtan who added methylglyoxal to honey samples and found that the addition of methylglyoxal (MGO) decreased the hydrogen peroxide percentage because of the inhibition effect of MGO on glucose oxidase (Majtan et al., 2014). Also we reported that honey samples with high vitamin C are associated with low hydrogen peroxide concentration which may be due to the antioxidant activity of vitamin C (Kerkvliet,1996). ...
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The altitude is well known to affect the temperature, parometric and oxygen pressure and the amount of UV radiation which affect the bees and the physicochemical properties of bee's honey. This study investigated the relations between the concentration of hydrogen peroxide, vitamin C and dicarbonyl molecules in honey samples from different floral origins and altitudes. Ten Ziziphus and twenty Acacia honey samples were collected directly from their bee farms. The hydrogen peroxide and vitamin C were measured using redox titartions while the dicarbonyl molecules concnetration was determined spectrophotometerically. The results were statisticaly analyzed by the ANOVA and t-test of the SPSS. The mean concentration of vitamin C in the Acacia and Ziziphus honey samples were 275.14± 82.3 and 239.16± 91.5 mg/100g, respectively. The mean hydrogen peroxide percentages in the Acacia and Ziziphus honey samples were 2.66± 0.81 and 4.94± 1.85%, respectively. The mean concentrations of the dicarbonyl molecules in the Acacia and Ziziphus were 324.62± 291.03 and 115.75± 94.9 mg/Kg, respectively. Significant variations were reported in the case of the hydrogen peroxide and the dicarbonyl molecules concentrations in the Acacia and Ziziphus honeys (p- values = 0.004 and 0.007, respectively). The altitude significantly afected the studied parameters. Honeys rich in dicarbonyl molecules have a high concentration of vitamin C and low content of hydrogen peroxide.