The controlled in vitro susceptibility of gastrointestinal pathogens to the antibacterial effect of manuka honey.

The susceptibility of common gastrointestinal bacteria against manuka honey with median level non-peroxide antibacterial activity (equivalent to that of 16.5% phenol) was investigated by determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) using a standardized manuka honey with the broth microdilution method. The measured sensitivity of bacteria showed that manuka honey is significantly more effective than artificial honey (a mixture of sugars as in honey), indicating that osmolarity is not the only factor that is responsible for the antibacterial activity of the honey. Most tested gastrointestinal pathogens have MIC and MBC values in the range of 5-10% of honey, other than Enterobacter spp. which was in the range of 10-17%. The difference in efficacy between the honey with and without hydrogen peroxide removed was also studied, and it was found that both hydrogen peroxide and the non-peroxide components contribute to the bacteriostatic and bactericidal activity of the honey. It was also found that treatment against multi-antibiotic resistant microorganisms such as Salmonella typhimurium DT104 and ESBL-producing organisms with manuka honey may be promising.

Effects on adhesion molecule expression and lymphocytes in the bovine mammary gland following intra-mammary immunisation.

Changes to adhesion molecule expression and lymphocyte populations were evaluated in alveolar mammary tissue collected from cows following an immunisation protocol that involved intra-mammary inoculation to induce an IgA response in mammary secretions. The right quarters of the udder were immunised; the left side acted as a control. Antibody titres in secretions showed that at least two animals responded with antigen-specific IgA. Numbers of T-lymphocytes were 4-fold higher in immunised glands compared with controls (P<0.05). IgA-, IgM- and IgG-positive cell numbers were significantly higher (P<0.01) in immunised glands compared with controls in three of the four cows. No mucosal addressin molecule-1 (MAdCAM-1), vascular cell-adhesion molecule-1 (VCAM-1) or peripheral node addressin (PNAd) protein expression was detected on smaller venules that stained positively for von Willebrand factor in alveolar mammary tissues, from either immunised or control glands. Both VCAM-1 and PNAd were detected on smaller venules in supramammary lymph nodes, however, there was no significant difference between immunised and control glands. Quantification of MAdCAM-1 mRNA showed very low expression in both immunised and control alveolar tissue compared with Peyer's patch positive-control tissue. These findings suggest that the bovine mammary gland is capable of a mucosal antibody response; however, MAdCAM-1 is not involved with lymphocyte homing to the mammary gland in this species.

The in vitro susceptibility of Campylobacter spp. to the antibacterial effect of manuka honey.

We report the antimicrobial effect of manuka honey against Campylobacter spp. isolated by a diagnostic laboratory from specimens from a community in New Zealand. The isolates were differentiated according to species level using multiplex PCR. C. jejuni (20 strains) and C. coli (7 strains) were identified. The clinical isolates identified and type culture collection strains of these species were subjected to testing to determine the minimum inhibitory concentration (MIC) of manuka honey using a microdilution technique. The MIC of the manuka honey against all of the Campylobacter tested was found to be around 1% (v/v) honey. The low MIC values suggest that honey might still inhibit the growth of campylobacteria after dilution by fluid in the gut, but the actual concentration of honey that can be achieved in the intestine is unknown. Therefore, clinical investigation is required to establish the efficacy of honey against Campylobacter spp. in the gut environment.

Adhesion molecule expression in the bovine mammary gland.

The bovine mammary gland requires lymphocytes for immune protection of the gland from foreign pathogens and, in addition, to transfer immune protection to the neonate via colostrum and milk. The process of homing primed lymphocytes to tissues is mediated by the interaction of cell-adhesion molecules displayed on the surface of lymphocytes and counter receptors displayed on the vascular endothelium. This study was conducted to identify the cell-adhesion molecules involved in homing lymphocytes to the bovine mammary gland at four different physiological stages; pregnant, colostral, lactation and involution. The expression and distribution of adhesion molecules in alveolar tissues and supramammary lymph nodes from the mammary glands of healthy cows was determined in situ by immunohistochemical analysis and compared with bovine Peyer's patch, used as a typical mucosal-associated lymphoid tissue and positive control. The mucosal addressin molecule, MAdCAM-1, was not detected in bovine mammary tissues at any of the four different physiological stages. Absence of MAdCAM-1 expression was verified by quantitative real-time RT-PCR analysis. Transcription levels of MAdCAM-1 mRNA were found to be more then 5 x 10(3)-fold lower in mammary alveolar tissues compared with bovine Peyer's patch tissues. In contrast to MAdCAM-1, phase-dependent protein expression of VCAM-1 was detected in both mammary alveolar tissues and the supramammary lymph nodes, with the highest expression observed in colostral phase cows. The protein expression in mammary alveolar tissues was limited to larger venules, although in colostral phase cows, VCAM-1 was also detected around the alveoli perimeter. In the supramammary lymph node, VCAM-1 protein was observed on both small and large venules. PNAd was detected in supramammary lymph nodes at all physiological stages of the mammary gland; however, it was not found in mammary alveolar tissues. Lymphocytes expressing beta7 were not detected in mammary tissues and lymphocytes expressing CD62L were only observed in the supramammary lymph nodes. Overall the data suggest that MAdCAM-1 and VCAM-1 are not involved in homing lymphocytes to the bovine mammary gland; whereas, VCAM-1 and PNAd may have this role in the supramammary lymph node.

The evidence supporting the use of honey as a wound dressing.

Some clinicians are under the impression that there is little or no evidence to support the use of honey as a wound dressing. To allow sound decisions to be made, this seminar article has covered the various reports that have been published on the clinical usage of honey. Positive findings on honey in wound care have been reported from 17 randomized controlled trials involving a total of 1965 participants, and 5 clinical trials of other forms involving 97 participants treated with honey. The effectiveness of honey in assisting wound healing has also been demonstrated in 16 trials on a total of 533 wounds on experimental animals. There is also a large amount of evidence in the form of case studies that have been reported. It has been shown to give good results on a very wide range of types of wound. It is therefore mystifying that there appears to be a lack of universal acceptance of honey as a wound dressing. It is recommended that clinicians should look for the clinical evidence that exists to support the use of other wound care products to compare with the evidence that exists for honey.

The antibacterial activity of honey against coagulase-negative staphylococci.

OBJECTIVES: Development of antibiotic-resistant strains of coagulase-negative staphylococci has complicated the management of infections associated with the use of invasive medical devices, and innovative treatment and prophylactic options are needed. Honey is increasingly being used to treat infected wounds, but little is known about its effectiveness against coagulase-negative staphylococci. The aim of this study was to determine the minimum active dilution of two standardized, representative honeys for 18 clinical isolates of coagulase-negative staphylococci. METHODS: An agar incorporation technique was used to determine the minimum active dilution, with dilution steps of 1% (v/v) honey [or steps of 5% (v/v) of a sugar syrup matching the osmotic effect of honey]. The plates were inoculated with 10 microL spots of cultures of the isolates. RESULTS: The honeys were inhibitory at dilutions down to 3.6 +/- 0.7% (v/v) for the pasture honey, 3.4 +/- 0.5% (v/v) for the manuka honey and 29.9 +/- 1.9% (v/v) for the sugar syrup. CONCLUSIONS: Typical honeys are about eight times more potent against coagulase-negative staphylococci than if bacterial inhibition were due to their osmolarity alone. Therefore, honey applied to skin at the insertion points of medical devices may have a role in the treatment or prevention of infections by coagulase-negative staphylococci.

The efficacy of honey in inhibiting strains of Pseudomonas aeruginosa from infected burns.

Because there is no ideal therapy for burns infected with Pseudomonas aeruginosa, there is sufficient need to investigate the efficacy of alternative antipseudomonal interventions. Honey is an ancient wound remedy for which there is modern evidence of efficacy in the treatment of burn wounds, but limited evidence for the effectiveness of its antibacterial activity against Pseudomonas. We tested the sensitivity of 17 strains of P. aeruginosa isolated from infected burns to two honeys with different types of antibacterial activity, a pasture honey and a manuka honey, both with median levels of activity. All strains showed similar sensitivity to honey with minimum inhibitory concentrations below 10% (vol/vol); both honeys maintained bactericidal activity when diluted more than 10-fold. Honey with proven antibacterial activity has the potential to be an effective treatment option for burns infected or at risk of infection with P. aeruginosa.

The sensitivity to honey of Gram-positive cocci of clinical significance isolated from wounds.

AIMS: To determine the sensitivity to honey of Gram-positive cocci of clinical significance in wounds and demonstrate that inhibition is not exclusively due to osmotic effects. METHODS AND RESULTS: Eighteen strains of methicillin-resistant Staphylococcus aureus and seven strains of vancomycin-sensitive enterococci were isolated from infected wounds and 20 strains of vancomycin-resistant enterococci were isolated from hospital environmental surfaces. Using an agar incorporation technique to determine the minimum inhibitory concentration (MIC), their sensitivity to two natural honeys of median levels of antibacterial activity was established and compared with an artificial honey solution. For all of the strains tested, the MIC values against manuka and pasture honey were below 10% (v/v), but concentrations of artificial honey at least three times higher were required to achieve equivalent inhibition in vitro. Comparison of the MIC values of antibiotic-sensitive strains with their respective antibiotic-resistant strains demonstrated no marked differences in their susceptibilities to honey. CONCLUSIONS: The inhibition of bacteria by honey is not exclusively due to osmolarity. For the Gram-positive cocci tested, antibiotic-sensitive and -resistant strains showed similar sensitivity to honey. SIGNIFICANCE AND IMPACT OF THE STUDY: A possible role for honey in the treatment of wounds colonized by antibiotic-resistant bacteria is indicated.

Potential of honey in the treatment of wounds and burns.

There has been a renaissance in recent times in the use of honey, an ancient and traditional wound dressing, for the treatment of wounds, burns, and skin ulcers. In the past decade there have been many reports of case studies, experiments using animal models, and randomized controlled clinical trials that provide a large body of very convincing evidence for its effectiveness, and biomedical research that explains how honey produces such good results. As a dressing on wounds, honey provides a moist healing environment, rapidly clears infection, deodorizes, and reduces inflammation, edema, and exudation. Also, it increases the rate of healing by stimulation of angiogenesis, granulation, and epithelialization, making skin grafting unnecessary and giving excellent cosmetic results.

Stimulation of TNF-alpha release in monocytes by honey.

Although evidence exists for the antibacterial effects of honey there is limited objective evidence for direct promotion of healing. We investigated the effect of manuka, pasture and an artificial honey on macrophage function. Reactive oxygen intermediate (ROI) production was assessed by luminol enhanced chemoluminescence and tumour necrosis factor-alpha (TNF-alpha) release was determined by immunoassay. ROI production was significantly (P<0.001) decreased by pasture honey and manuka honey. TNF-alpha release was significantly enhanced (P<0.001) in unprimed MM6 cells by manuka and pasture honey but was not altered in primed cells. These results could explain the suggested therapeutic properties of honey in promoting wound healing.

The potential of honey to promote oral wellness.

Honey has been used as a medicine throughout the ages and in more recent times has been "rediscovered" by the medical profession for treatment of burns, infected wounds, and skin ulcers. The large volume of literature reporting its effectiveness indicates that honey has potential for the treatment of periodontal disease, mouth ulcers, and other problems of oral health.

Antibacterial activity of honey against strains of Staphylococcus aureus from infected wounds.

The antibacterial action of honey in infected wounds does not depend wholly on its high osmolarity. We tested the sensitivity of 58 strains of coagulase-positive Staphylococcus aureus, isolated from infected wounds, to a pasture honey and a manuka honey. There was little variation between the isolates in their sensitivity to honey: minimum inhibitory concentrations were all between 2 and 3% (v/v) for the manuka honey and between 3 and 4% for the pasture honey. Thus, these honeys would prevent growth of S. aureus if diluted by body fluids a further seven-fold to fourteen-fold beyond the point where their osmolarity ceased to be completely inhibitory. The antibacterial action of the pasture honey relied on release of hydrogen peroxide, which in vivo might be reduced by catalase activity in tissues or blood. The action of manuka honey stems partly from a phytochemical component, so this type of honey might be more effective in vivo. Comparative clinical trials with standardized honeys are needed.

Effect of testosterone on binding of insulin-like growth factor-I (IGF-I) and IGF-II in growing antlers of fallow deer (Dama dama).

Testosterone regulation of antler growth may be via the insulin-like growth factors (IGFs). Using histological autoradiography we have measured the specific binding of IGF-I and IGF-II to antler sections during normal growth and during the maturation which follows testosterone treatment of adult fallow deer. In antlers from 20 to 100 days following casting, IGF-I binding was constant within each histological region until 80 days. Between this time and 100 days there was decreased binding to chondrocytes (P < or = 0.01) and increased binding to the reserve mesenchyme/perichondrium (P < or = 0.001). Following testosterone treatment, IGF-I binding declined in dermis (P < or = 0.05), reserve mesenchyme/perichondrium (P < or = 0.05), and chondroblasts (P < or = 0.01). Specific binding of IGF-II showed no change during normal or testosterone-stimulated growth. In conclusion, the regulation of antler maturation by testosterone may include IGF action, probably via the Type 1 IGF receptor.