Thin-layer chromatography in the authenticity testing of bee-products.

Quality control, nutritional value and the monitoring of hazardous residues in honey bee- products have become major topics for both producers and consumers. Due to its potential role in human health, bee-products rich in bioactive compounds are becoming increasingly popular. This review aims to provide an overview of thin-layer chromatography methods used in quality control,authenticity testing and chemical profiling of bee-products in order to help scientists engaged in the field of bee-products chemistry to utilize the advantages of this technique in the detection and elimination of fraudulent practices in bee-product manufacturing. Recently, hyphenation of thin-layer chromatography, image analysis and chemometrics support bee-products analysisbysimultaneousdeterminationofanalytes with different detection principles, identification of individual bioactive compounds as well as structure elucidation of compounds. Highlighted opportunities of thin-layer chromatography could encourage further investigations that would lead to improvements in the detection and elimination of marketing fraudulent practices.

Treatment of Dehisced, Thoracic Neonatal Wounds With Single-Use Negative Pressure Wound Therapy Device and Medical-Grade Honey: A Retrospective Case Series.

PURPOSE: The purpose of this study was to report on our experience with a portable, single-use negative pressure wound therapy device used in combination with activated active Leptospermum honey (ALH) in the treatment of colonized or infected, dehisced, thoracic wounds in neonates with complex congenital heart disease. DESIGN: Retrospective, descriptive study. SUBJECTS AND SETTING: We reviewed medical records of 18 neonates and reported on findings from 11; the remaining 7 were not included secondary to incomplete records, transfer to a different institution prior to wound healing, or death. The median age of our patients was 12 days (range, 2 days to 5 weeks); their mean gestational age was 34 weeks. All of the neonates had acquired postoperative wound dehiscence that were colonized or infected and were treated in the neonatal intensive care unit (NICU) at Cohen Children's Medical Center (New Hyde Park, New York), a regional perinatal center with a level 4 NICU. METHODS: Wound cultures were obtained on all patients prior to treatment commencement. All cultures were repeated on day 4 of treatment. Systemic antibiotics were administered as necessary. No complications were observed related to the use of negative pressure wound therapy device and ALH. All patients were followed until discharge home or transfer to another facility. The pain scores during placement and removal were acceptable (between 1 and 3; median = 2) using the Neonatal Infant Pain Scale. Staff and parents indicated that the combination of ALH and the negative pressure wound therapy device did not interfere with daily care and parental bonding. CONCLUSIONS: Use of ALH and a single-use negative pressure wound therapy device was successful in this series of 11 neonates with complex congenital heart disease.

Honey yield of different commercial apiaries treated with Lactobacillus salivarius A3iob, a new bee-probiotic strain.

The main objective of this study was to determine the impact of Lactobacillus salivarius A3iob, a honey bee gut-associated strain (GenBank code access KX198010), on honey yield. Independent assays were conducted from May to September 2014 and 2015, in three commercial apiaries: Tilquiza, El Carmen and Yala, all located in north-western Argentina. Local Apis mellifera L. bees were kept in standard Langstroth hives; treated hives were fed once a month with 1×105 cfu/ml viable Lactobacillus cells, administered to the bees through a Doolittle-type feeder in 125 g/l sucrose syrup. Control hives were only given the syrup mixed with MRS sterile broth. The main honey harvest was done in December in all groups and we found that there was an overall increase in honey yield from the treated hives. In 2014, all treated hives produced between 2.3 to 6.5 times more honey than the controls. However, in 2015, higher honey average yields in the treated hives at El Carmen and Yala were obtained, yet not at Tilquiza, because of a slight mishap. They experienced the swarming of several bee colonies due to a higher number of bees without appropriate management, which caused the control group to yield more honey compared to the hives fed with Lactobacillus. Interestingly, at El Carmen, two honey harvests were recorded: one in winter and another in summer (July and December 2015, respectively). This unexpected result arose from the particular flora of the region, mainly Tithonia tubaeformis, which blooms in winter. L. salivarius A3iob cells prove to be a natural alternative that will positively impact the beekeepers' economy by providing a higher honey yield.

Composition, labelling, and safety of food supplements based on bee products in the legislative framework of the European Union - Croatian experiences.

The European Union market is overflown by food supplements and an increasing number of consumers prefer those where bee products play an important part in their composition. This paper deals with complex European Union legislation concerning food supplements based on bee products, placing a special emphasis on their composition, labelling, and safety. Correct labelling of food supplements also represents a great challenge since, in spite of legal regulations in force, there are still open issues regarding the statements on the amount of propolis, which is not clearly defined by the legal framework. One of the key issues are the labels containing health claims from the EU positive list approved by the European Food Safety Authority. Emphasis will also be placed on informing consumers about food, as statements which imply the healing properties of food supplements and their capacity to cure diseases are forbidden. One of the key elements of product safety is HACCP based on the EU Regulations EC 178/02 and 852/2004. Health safety analyses of food supplements with bee products used as raw materials, which are standardised by legal regulations will also be discussed. In the future, attention should also be paid to establishing the European Union "nutrivigilance" system. Croatian experiences in addressing challenges faced by producers, supervisory entities, and regulatory and inspection bodies may serve as an example to countries aspiring to become part of the large European family.

A suggestion for royal jelly specifications.

This article proposes guidelines for quality standards of royal jelly. The proposals are based on two sets of data; the first from our study of the factors that may affect the royal jelly's chemical composition (protein and sugar supplementation of beehives) and the second on the analysis of a great number of samples from across Greece to establish natural variability of this product. We compared our findings with the adopted national limits, the proposals of the working group of the International Honey Commission (IHC), and the draft proposal of the International Organization of Standardization (ISO). The studied parameters included moisture, total proteins, sugars (fructose, glucose, sucrose, total sugars), and 10-hydroxy- 2-decenoic acid (10-HDA). Our results indicate that the limits for royal jelly in some countries should be amended and the proposals of the IHC and the ISO reviewed in view of recent data on variability. We believe that our proposals could be considered for setting global standards for royal jelly, as they incorporate national legislations, proposals of scientific groups, experimental data, and updated information.

Comprehensive study of honey with protected denomination of origin and contribution to the enhancement of legal specifications.

In this study the characterization of a total of 60 honey samples with Protected Denomination of Origin (PDO) collected over three harvests (2009-2011, inclusive), from the Northeast of Portugal was carried out based on the presence of pollen, physicochemical and microbiological characteristics. All samples were found to meet the European Legislation, but some didn't meet the requirements of the PDO specifications. Concerning the floral origin of honey, our results showed the prevalence of rosemary (Lavandula pedunculata) pollen. The microbiological quality of all the analyzed samples was satisfactory, since fecal coliforms, sulfite-reducing clostridia and Salmonella were absent, and molds and yeasts were detected in low counts. Significant differences between the results were studied using one-way analysis of variance (ANOVA), followed by Tukey's HSD test. The samples were submitted to discriminant function analysis, in order to determine which variables differentiate between two or more naturally occurring groups (Forward Stepwise Analysis). The variables selected were in this order: diastase activity, pH, reducing sugars, free acidity and HMF. The pollen spectrum has perfect discriminatory power. This is the first study in which a honey with PDO was tested, in order to assess its compliance with the PDO book of specifications.

Honey and wound healing: an overview.

Honey has been used to treat wounds throughout the ages. This practice was rooted primarily in tradition and folklore until the late 19th century, when investigators began to characterize its biologic and clinical effects. This overview explores both historic and current insights into honey in its role in wound care. We describe the proposed antimicrobial, immunomodulatory, and physiologic mechanisms of action, and review the clinical evidence of the efficacy of honey in a variety of acute and chronic wound types. We also address additional considerations of safety, quality, and the cost effectiveness of medical-grade honeys. In summary, there is biologic evidence to support the use of honey in modern wound care, and the clinical evidence to date also suggests a benefit. However, further large, well designed, clinical trials are needed to confirm its therapeutic effects.

Honey promotes angiogeneic activity in the rat aortic ring assay.

OBJECTIVE: To investigate possible effects of honey on angiogenesis, using in vitro analogues of angiogenesis and an endothelial proliferation assay. METHOD: Using an in vitro rat aortic ring assay we compared pseudotubule formation by medicinal honey (Activon), supermarket honey (Rowse) and a honey-based ointment (Mesitran), with that of artificial honey (70% w/w sugar glucose/fructose). Pseudotubules were analysed using TCS Cellworks AngioSys software. The Angiokit sytem was used to validate the results. Using the MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium. Bromide] assay, toxicity was also assessed on human umbilical vein endothelial cells (HUVEC) directly adherent to plastic. RESULTS: All honey preparations stimulated pseudotubule formation, maximal at around 0.2% honey. Medicinal honeys were more active than Rowse. The effect was not attributable to the sugar content. Among the honeys tested, the Manuka-based Activon preparation reduced residual viable biomass compared with a sugar control at > 0.32% v/v concentration. Rowse had a similar effect only at 2.5%, the highest dose tested. CONCLUSION: The influence of honey constituents on angiogenesis in a wound dressing context is likely to be positive, but would depend on the effective dilution of the honey and the penetration of the active constituents against an osmotic gradient. The extent to which this occurs has yet to be established. CONFLICT OF INTEREST: This work was conceived, designed and executed by the authors. Medical honey preparations were supplied unconditionally but free of charge by the distributors.

Physicochemical, microbiological and antimicrobial properties of commercial honeys from Portugal.

The present study aimed to characterize five commercial honeys available in the Portuguese market in respect to their floral origins, physicochemical parameters and microbial safety and commercial quality assessment. Pollen profile, colour, moisture content, ash, acidity, electrical conductivity, pH, reducing sugars, apparent sucrose and HMF were the parameters analysed in each honey sample. Aerobic mesophiles, moulds and yeasts, fecal coliforms and sulphite-reducing clostridia were the microbial contaminants of interest studied. The antimicrobial effect against four fermentative yeasts was determined. Concerning the physicochemical parameters, all honey samples were found to meet European Legislation (EC Directive 2001/110) for all parameters, except for HMF and apparent sucrose. Microbiologically, the commercial quality was considered good and all samples showed to be negative in respect to safety parameters. We also verified that the presence of honey differentially affected the growth of fermentative yeasts under study, depending on the type of yeast, but this growth was not significantly influenced by the type of honey used.

Antibacterial activity of bee honey and its therapeutic usefulness against Escherichia coli O157:H7 and Salmonella typhimurium infection.

The authors studied the effect of storage period and heat on the physical and chemical properties of honey and proceeded to study the antibacterial effect of honey on Escherichia coli and Salmonella typhimurium. In samples of honey (Egyptian clover honey) that were heat-treated and stored over a long period of time, water content decreased, hydroxymethyl furfural (HMF) was produced and increased in concentration, and enzyme activity decreased. Colour, measured in optical density, was markedly affected in honey samples stored over long periods of time, as was the refractive index, but electrical conductivity remained unaffected by storage or heating. Similarly, the storage period had no effect on pH value. To study the therapeutic effect of honey on E.coli and S. typhimurium, 25 isolates of E. coil O157:H7 (18.5%) and 49 isolates of S. typhimurium (36.2%) were isolated from 135 samples taken from children and calves (30 stool samples from children and 105 samples from calf organs and faecal swabs). Most E. coli O157:H7 and S. typhimurium isolates were highly resistant to most antibiotic discs. In vitro, the antibacterial effect of honey was more pronounced on E. coil O157:H7 than on S. typhimurium. Water content, pH value, HMF and the presence of H2O, all played an important role in the potency of olover honey as an antibacterial agent. In vivo, mice were used as a model for studying the parenteral usefulness of honey as an antibacterial agent against both pathogens. The antibacterial activity of honey that had been stored over a long period of time decreased and high concentrations of honey proved more effective as antibacterial agents. In this study there was lower mortality among mice treated with honey but the parenteral application of honey and its therapeutic properties require further investigation.