Artificial Honey Research Articles

Physicochemical and Microbiological Analysis of Stingless Bees Honey Collected from Local Market in Malaysia

The growing demand for honey in the market has led to the occurrence of the tampering honey with foreign substances and increases the production of artificial honey. Due to this concern, this study works on the physicochemical and microbial characterization of stingless bee honey. The physicochemical analysis showed that the honey possessed pH (2.51–3.26), free acidity (121.1 to 318.7 meq/kg), moisture (19.4–30.9%), electrical conductivity (0.33–0.69 mS/cm), ash content (2.75–4.31 g/100g), Hydroxymethylfurfural (HMF) content (35.4 to 461.7 mg/kg) and diastase activity (2.71 to 6.11 DN). Also, sugar profile of honey showed that the honey contained fructose (15.03–32.52 g/100g), glucose (12.17–34.55 g/100g) and sucrose (0.01–7.29 g/100g). The harvested honey, H1, and H2 have the highest potential to become an antibacterial agent to treat disease compared to commercial honey samples because they were active against Gram-negative bacteria. All analyzed samples were within the maximum limit of the quality criteria set by the Malaysian Kelulut Standard and Codex Alimentarius except for free acidity, HMF, and Diastase Number. All the data obtained is vital in order to create a specific statute for stingless bees honey in Malaysia that may help to protect the consumer from purchasing adulterated honey.

The effect of different types of honey on the lipid profile of streptozotocin-induced diabetic rats.

IntroductionThe effect of honey consumption in diabetic patients has been contradictory. The aim of the present animal study was to compare the effect of different types of honey on the lipid profile in diabetic rats.Material and methodsSixty-four male Wistar rats were divided into two main groups: a streptozotocin-induced diabetes mellitus (DM) group (including four subgroups) and a healthy group (including four subgroups), based on random allocation. Three subgroups of each main group were given 1 mg/kg of three different types of honey (acacia, astragalus, and artificial honey) by oral gavage for 10 weeks. The control groups were given distilled water. Blood samples were collected, and the lipid profile was measured and compared between the eight groups after the intervention.ResultsThe levels of LDL, triglycerides (TG), and total cholesterol (Tchol) in DM rats treated with astragalus honey were significantly lower and the HDL level was significantly higher compared to the other DM and healthy groups (all p-values < 0.05). LDL, TG, and Tchol levels in DM rats treated with artificial honey were significantly higher, and HDL levels were significantly lower than for other types of honey and for the control groups (all p-values < 0.05). LDL, HDL, TG, and Tchol levels in healthy rats were not significantly different between the groups (p-value > 0.05).ConclusionsDifferent types of honey (acacia, astragalus, and artificial honey) had various effects on serum lipid profiles in diabetic rats. The results of this study indicated that the effect of honey on diabetic patients can vary widely based on its source.

The monoclonal antibody for discrimination of natural honey against artificial honey.

BackgroundHoney is a natural product used as food, medicine, or cosmetics for very long time and is made by bees. Honey contains various components such as sugar, protein, minerals, and vitamins. Honey is made by Apis cerana or Apis mellifera, which commonly has major royal jelly proteins (MRJPs) as a major protein. To discriminate between natural honey (NH) and artificial honey (AH), many researchers tried method of physicochemical analysis. However, the analysis results were ambiguous and not stable.ResultsWe have produced a monoclonal antibody that recognizes MRJPs of honeys in common. Monoclonal antibody has advantage such as accuracy, sensitivity, and stability as the standard. The specificity and affinity of produced antibody were measured by western blotting and enzyme‐linked immunosorbent assay. As a result, this monoclonal antibody specifically recognized MRJPs of NH and did not recognize AH which has not including MRJPs.ConclusionNatural honey could be able to distinguish from AH accurately by using this monoclonal antibody. Also, this method could be commercially applicable.

A fusion fuzzy model for detecting phony accounts in social networks

In online social network’s phony account detection is one of the major task among the ability of genuine user from forged user account. The fundamental objective of detection of phony account framework is to detect fake account and removal technique in Social network user sites. This work concentrates on detection of phony account in which it depends on normal basis framework, transformative Algorithms and fuzzy technique. Initially, the most essential attributes including personal attributes, comparability techniques and various real user review, tweets, or comments are extricated. A direct blend of these attributes demonstrates the significance of each reviews tweets comments etc. To compute closeness measure, a consolidated strategy in view of artificial honey bee state Algorithm and fuzzy technique are utilized. Second approach is proposed to alter the best weights of the normal user attributes utilizing the social network activities/transaction and inherited Algorithm. Finally, a normal rank rationale framework is utilized to calculate the final scoring of normal user activities. The decision making of proposed approach to find phony account are variation with existing techniques user behavioral analysis using data sets and machine learning techniques such as crowdflower_sample and genuine_accounts_sample dataset of facebook and Twitter. The outcomes demonstrate that proposed strategy overcomes the previously mentioned strategies.

The contribution of α-dicarbonyl compound dependent radical formation to the antiseptic effect of honey

Honey is known for its wound healing potential. In this study we investigated free radical formation and inhibition in different honeys and the influence on bacterial growth inhibition. α-Oxaldehydes and free amino acids were shown to be present in manuka and non-manuka honeys. With electron spin resonance we measured that methylglyoxal (MG) and glyoxal (GO) form different carbon centered radicals under the influence of either hydrogen peroxide or arginine and lysine. These radicals were also formed in a combination of α-oxaldehydes and amino acids in artificial honey, which was quantified by the cytochrome C assay. Also, a strong bactericidal activity for MG and GO (minimum concentration 0.6 and 1.25 mM) was shown against different bacterial strains. Interestingly, honeys with a high MG content were shown to be better free radical scavengers for hydroxyl and superoxide radicals, implying a complex mechanism for free radical donation and scavenging in honey.

Characterization of Apis mellifera Honey of Different Botanical and Geographical Origins in Egypt

Seven honey types identified botanically in terms of their floral origin (clover, Cedrus, citrus, banana, cotton, Brazilian pepper and sun flower), and their geographical regions during 2016-2017 have been characterized in comparison with artificial honey. Characterization of honeys was based on their physicochemical properties, antibacterial & antifungal activity, heavy metal contents and multi-pesticides residues using quantitative analysis methods. All honeys tested were natural and give negative results for adulteration tests. Sun flower and cotton honeys showed the highest sucrose content regardless the artificial honey sucrose content (74 ±1 % (g/100 g). The water content and pH value of different honeys investigated were within the standard limits 20g/100g and 3.4-6.1, respectively. Hydroxymethylfurfural (HMF) content in most honeys was below the maximum allowable limit (MAL) (40 mg/kg) in honey, except for banana honey. None of the honeys tested showed any antifungal activity. Artificial honey showed no growth inhibition against reference strains of bacteria and fungi tested. Metal content of Fe, Zn, Pb, Cu in Egyptian honeys fulfill the (MAL) described in the standard codex for honey except for Cd with concentrations exceeding the MAL (0.05 mg/kg) in most types of honey, except for artificial honey. Malathion, chlorpyrifos and tau-fluvalinate were the most frequently detected (25%, n=2) insecticides in honey samples. It can be concluded that the best Egyptian honeys tested in terms of antibacterial activity were Brazilian pepper honey followed by Cedrus honey. The data obtained are extremely important for the public, health officials for medical and nutritional applications of honey.

Anatolian honey is not only sweet but can also protect from breast cancer: Elixir for women from artemis to present.

Natural products with bioactive components are widely studied on various cancer cell lines for their possible cytotoxic effects, recently. Among these products, honey stands out as a valuable bee product containing many active phenolic compounds and flavonoids. Numerous types of multifloral honey and honeydew honey are produced in Turkey owing to its abundant vegetation. Therefore, in this study, we investigated the cytotoxic effects of particular tree-originated honeys from chestnut, cedar, pine, and multifloral honey on cell lines representing different types of the most common cancer of women, breast cancer, MCF7, SKBR3, and MDAMB-231, and fibrocystic breast epithelial cell line, MCF10A as a control. All honey samples were analyzed biochemically. The dose- (1, 2.5, 5, 7.5, and 10 µg/mL) and time (24th, 48th, and 72nd hours)-dependent effects of ethanol/water solutions of the honey samples were scrutinized. Cell viability/cytotoxicity was evaluated by the water soluble tetrazolium Salt-1 (WST-1) method. Apoptotic status was detected by Annexin V-PI assay using FACSCalibur. The statistical analysis was performed using GraphPad Prism 6 and the clustering data analysis with the R programming language. The biochemical analyses of the honey samples showed that the tree-originated honey samples contained more total phenolic compounds than the multifloral honey. Phenolic content of the honey types increases in order of multifloral, pine, cedar, and chestnut, respectively, which is compatible with their cytotoxic affectivity and dark color. In addition, the antioxidant capacity of the studied honey types was observed to increase in order of multifloral < pine < cedar ≅ chestnut. According to the WST-1 data, chestnut honey induced cytotoxicity over 50% on all the cell lines, including the control MCF10A cells, even with low doses (honey concentrations starting from 1 µg/mL) (P < 0.0001). Similarly, Cedar honey was observed to be the second most effective honey in this study. Cedar honey, with the dose of 1 µg/mL, was detected statistically highly significant on MCF10A, MCF7, and SKBR3. In contrast, pine honey showed dramatically significant cytotoxicity only on the MDAMB 231 cells with a 1 µg/mL dose at the same time point (P = 0.018). While pine honey caused an anticancer effect on the MCF-7 and SKBR3 cancer cell lines with a 2.5-5 µg/mL dose (P < 0.0001), like cedar and chestnut honeys, it increased the viability of the MCF10A control cells with the doses of 2.5-5 µg/mL. It only showed cytotoxicity with higher doses (10 µg/mL) on the MCF10A cell line (P < 0.0001). Moreover, we have observed that the multifloral and artificial honey samples were mostly ineffective or increased cell viability with the doses of 1-5 µg/mL. Apoptotic effects of the other honey samples on the MCF-7 cell line were found as chestnut> pine> cedar> multifloral in the Annexin V-propidium iodide (PI) analysis. Chestnut, cedar, and pine honey displayed a remarkably cytotoxic effect on breast cancer cell lines, MCF7, SKBR3, and even on the most aggressive MDAMB 231, representing the triple negative breast cancer, which lacks of targeted anticancer therapy. The chestnut and cedar honeys stand out to be the most cytotoxic on all cell lines, while pine honey was found to be the least toxic on control cells with appropriate toxicity on the cancer cells. © 2017 IUBMB Life, 69(9):677-688, 2017.

An Exploration for the Analysis of Ground Water Possibility in Case Based Recommendation System using Hybrid Cuckoo Search and Artificial Honey Bee Algorithm

Groundwater Possibility Exploration is one of the prime problems in human life. Human and other social species are directly or indirectly dependent on water resources. But with increasing population and instant changing environmental conditions, groundwater resources are also becoming less day by day. One approach of manual exploration can be used but that is most laborious and time consuming task. So, there is need of some autonomous approach with which we can find the more groundwater resources without digging the bore well. This paper presents an autonomous approach in case based recommendation system using hybrid cuckoo search and artificial honey bee algorithm for groundwater possibility detection. The proposed concept is structured in the manner to have the input in the form six attributes of slope, geology, landuse, lineament, soil & landform and give output in the form of low, intermediate and high possibility. In this method, case based reasoning used in the manner to retrieve the previous knowledge of use cases. The cases are actually the host bird’s nest and input is cuckoo’s egg. CS originates from the behaviour of certain species of cuckoo which lay eggs in other birds nest in parasitic manner. If cuckoo’s egg adapts the behaviour of host nests it will exist otherwise the host bird will discard cuckoo’s egg. The minimum similarity value is calculated using ABC algorithm. CBR life cycle further uses the information from this metaphor. The overall method is evaluated using the parameters of sensitivity, specificity and accuracy.

Kinetics of conversion of dihydroxyacetone to methylglyoxal in New Zealand mānuka honey: Part IV – Formation of HMF

During a study of the conversion of dihydroxyacetone (DHA) to methylglyoxal (MGO) in maturing New Zealand mānuka honey, the kinetics of formation of 5-(hydroxymethyl)furfural (HMF) was studied at temperatures from 4 to 37°C. Formation of HMF was first-order during an induction period and zero-order thereafter indicating that the mechanism includes the formation of certain critical intermediates and that these require time to build up; the duration of the induction period depended primarily upon temperature.The zero-order rate constant at 37°C was the same for mānuka honey and clover honey doped with 2000 or 10,000mg/kg DHA and for artificial honey with 2000mg/kg of DHA and either alanine or proline and alanine added. Zero-order rate constants for artificial honey with added amino acids were less than for a control without amino acids.A simulation was created to predict the formation of HMF over time at 37°C in mānuka honey.

Analysis of Dibutyl Phthalate and Oleamide in Stingless Bee Honey Harvested from Plastic Cups

Increasing demands for stingless bee honey in Malaysia have prompted alternative method using plastic cups as artificial honey pots within beehives to enhance yield. However, honey’s acidity may lead to migration of plastic additives, affecting body’s physiological responses on prolonged consumption. This study was performed to determine the presence of dibutyl phthalate (DBP) and oleamide in stingless bee honey collected from plastic cups. A method for simultaneous extraction and detection of both analytes by solvent terminated dispersive liquid-liquid microextraction (ST-DLLME) coupled with high performance liquid chromatography (HPLC) was developed. Good linearity was observed with coefficient of determination R2 ≥ 0.97 in the concentration range of 0.5-50 and 10-250 μg/g for DBP and oleamide, respectively. The limit of detection for DBP was 0.15 μg/g; while oleamide was 4 μg/g. The limit of quantitation for DBP and oleamide was 0.5 and 11 μg/g, respectively. DBP and oleamide were not detected in all the honey samples collected.

Trypsinized Cry1Fa and Vip3Aa have no detrimental effects on the adult green lacewing Chrysopa pallens (Neuroptera: Chrysopidae)

An exposure bioassay was established for green lacewing Chrysopa pallens (Rambur) (Neuroptera: Chrysopidae) adults using a suitable artificial diet and honey–water solution (20%) to assess the toxicity of trypsinized Cry1Fa and Vip3Aa. Lethal and sub-lethal life table parameters were unaffected after C. pallens adults were given honey–water solutions containing Cry1Fa and Vip3Aa (50 µg/ml) and an artificial diet. In contrast, life table parameters of C. pallens adults were significantly affected when boric acid was mixed with the honey–water solution as a positive control. The uptake, temporal stability and bioactivity of Cry1Fa and Vip3Aa before and after C. pallens access to honey–water were confirmed using double antibody sandwich enzyme-linked immunosorbent assays and a bioactivity verification bioassay. The results prove that trypsinized Cry1Fa and Vip3Aa are safe for C. pallens adults, thus it is speculated that transgenic crops expressing Cry1Fa and Vip3Aa have no detrimental effects on lacewings and are compatible with biological control programs. This study describes a robust experimental design for evaluating the potential toxicity of alkaline gut-activated Bacillus thuringiensis (Bt) proteins on C. pallens adults which can be used to determine the potential toxicity of other Bt proteins on this species.

Tingkat kesukaan lebah madu Apis Mellifera L. terhadap pakan tambahan campuran madu mengkudu

Feeding supplementary food containing nony fruit juice was tested on honeybee colonies in honey producing . The aims of this research were (1) to determine the preference level of honeybees to the supplementary food containing honey and nony mixture, (2) to identify the organoleptic properties of the produced honeyessence of nony, (3) to determine fiber content of produced honey-essence of nony, and (4) to identify the colours of honey produced. The design used in the experiment was Latin Square Design. There were four treatments namely each concentration : A (0%) as control, B (12,5%), C (25%) dan D (37,5%) in dilution of supplementary food an amount of 600 grams wich consist of nony fruit juice and water. Each composition of dilution were A (300, 0, 300), B (225, 75, 300), C (150, 150, 300) dan D (75, 225, 300) . Every one replication was done for six days with six day interval. The result were as follows: The higher concentration of nony juice in the food suplement the smaller food were taken. The differences in the olour, the flavour and the taste of the artificial honey didn’ depend on the amount of the food suplement taken honey bees, but depend on nony fruits juice concentration in the honey produced. The higher concentration of nony fruit juice in the artificial honey produced the darken in colour, sour in taste and the weaken in flavour. Feeding with food suplement containing nony juice increases the fiber content in the honey.

Leptosperin is a distinct and detectable fluorophore in Leptospermum honeys

New Zealand manuka (Leptospermum scoparium) honey exhibits two unique fluorescence signatures that distinguish it from other honey types. One of these is the MM1 fluorescence marker (270–365nm excitation–emission) which we show is due to a Leptospermum nectar-derived compound, leptosperin. Synthetic or honey-purified leptosperin not only displayed an identical fluorescence spectrum, but supplementation of leptosperin into clover or artificial honeys generated the MM1 fluorescence signature. There was a quenching effect of the honey matrix on leptosperin fluorescence but otherwise leptosperin was chemically stable over prolonged storage at 37°C. Leptosperin was also present in the woody-fruited Australian Leptospermum species at elevated concentrations but virtually absent in Leptospermum subtenue suggesting its elevated expression developed following the mid-Miocene separation of the genus. These findings suggest that fluorescence spectroscopy could offer a rapid and high-throughput screening method for identification of Leptospermum honeys using the MM1 fluorescence marker.

Quantification of bee-derived peptide defensin-1 in honey by competitive enzyme-linked immunosorbent assay, a new approach in honey quality control

We established and evaluated a polyclonal antibody based competitive enzyme-linked immunosorbent assay for the quantification of defensin-1 in honey. The assay showed an inhibitory concentration (IC&lt;sub&gt;50&lt;/sub&gt;) value of 111.5 ± 15.41 ng/ml with a detection limit of 7.8125 ng/ml. The regaining of defensin-1 in spiked ‘artificial honey’ was between 87.05 and 112.96% with relative standard deviation less than 9.2%. Sensitivity and specificity of the test were experimentally validated on a sample of 20 different honeys. The antibacterial activity of these honey samples showed a significant concentration-dependent correlation with the production of defensin-1 (n = 20; r = −0.6598; P = 0.0016). The assay provides a specific and sensitive method for the screening of defensin-1 in honey. The method to detect honeybee-derived proteins in honey is a promising approach to verifying the authenticity of honey. The defensin-1 ELISA could also be used for the rapid screening of honeys suitable for medicinal purposes.

한국산 벌꿀의 밀원별 단당, 이당 및 삼당류의 정량 특성

Sugar profiles of 45 Korean honey samples (15 acacia, 15 multi-floral, 10 chestnut, and 5 artificial honey samples), which are commercially available in the Korean markets, were analyzed using gas chromatography/mass spectrometry (GC/MS) through TMS-oxime and TMS-methoxime derivatization. The average invert sugar contents in acacia, multi-floral, chestnut, and artificial honey samples were 71.2±1.05, 68.7±3.26, 63.2±1.85, and 68.0±2.10%, respectively. Fourteen disaccharides were detected from the samples, and the average content of major disaccharides was higher in order of turanose, maltulose, maltose, trehalulose, kojibiose, isomaltose, and nigerose. The average content of total disaccharides was highest in chestnut and lowest in acacia. Seven trisaccharides were detected from the samples, and the average content of trisaccharides was the highest in artificial honeys, which had high erlose content. The total content of disaccharides and trisaccharides was highest (16.0±2.03%) in chestnut honey and lowest (9.70±1.75%) in acacia honey.

Kinetics of conversion of dihydroxyacetone to methylglyoxal in New Zealand mānuka honey: Part III – A model to simulate the conversion

A kinetic model for the conversion of dihydroxyacetone (DHA) to methylglyoxal (MGO) in honey is proposed; a building block approach was used to create the model. Artificial honeys doped with DHA and individual perturbants were fitted first, then multiple perturbants (alanine, proline and iron, and combinations of these) were fitted before comparing the simulation to real honey samples (doped clover and mānuka honey). The main responses in the prediction model were DHA, MGO, proline, primary amino acids, acidity, 3-phenyllactic acid and 4-methoxyphenyllactic acid. Three temperatures (20, 27 and 37°C) were studied and the conversion of DHA to MGO was monitored over at least 1year. Differences in the conversion between clover doped with DHA and mānuka honey were observed. The simulation fitted well for the honeys tested.

Kinetics of the conversion of dihydroxyacetone to methylglyoxal in New Zealand mānuka honey: Part II – Model systems

The irreversible dehydration reaction of dihydroxyacetone (DHA) to methylglyoxal (MGO) in a honey model system has been examined to investigate the influence of added perturbant species on the reaction rate. The secondary amino acid proline, primary amino acids (alanine, lysine and serine), and iron, or combinations of these perturbants, were added to artificial honey with either DHA or MGO and stored at 20, 27 and 37°C. These systems were monitored over time. A 1:1 conversion of DHA to MGO was not observed in any system studied, including the control system with no added perturbants. Addition of proline to the matrix increased consumption of DHA but did not produce any more MGO than the control sample. Lysine and serine behaved similarly. Alanine enhanced the conversion of DHA to MGO and had the best efficiency of conversion of DHA to MGO for the amino acids studied. An iron II salt enhanced the conversion of DHA to MGO, even in the presence of proline.

Efficiency of Polyphenol Extraction from Artificial Honey Using C18 Cartridges and Amberlite® XAD-2 Resin: A Comparative Study

A comparative study of the extraction efficiency of nine known polyphenols [phenolic acids (benzoic acid, dihydroxybenzoic acid, gallic acid, trans-cinnamic acid, and vanillic acid) and flavonoids (naringenin, naringin, quercetin, and rutin)] was conducted by deliberately adding the polyphenols to an artificial honey solution and performing solid phase extraction (SPE). Two SPE methods were compared: one using Amberlite XAD-2 resin and another one using a C18 cartridge. A gradient high performance liquid chromatography system with an RP18 column and photodiode array detector was utilized to analyze the extracted polyphenols. The mean percent of recovery from the C18 cartridges was 74.2%, while that from the Amberlite XAD-2 resin was 43.7%. The recoveries of vanillic acid, naringin, and rutin were excellent (&gt;90%); however, gallic acid was not obtained when C18 cartridges were used. Additionally, the reusability of Amberlite XAD-2 resin was investigated, revealing that the mean recovery of polyphenols decreased from 43.7% (1st extraction) to 29.3% (3rd extraction). It was concluded that although Amberlite XAD-2 resin yielded a higher number of compounds, C18 cartridges gave a better extraction recovery. The lower recovery seen for the Amberlite XAD-2 resin also cannot be compensated by repeated extractions due to the gradual decrease of extraction recovery when reused.

Identification and Quantitation of 2-Acetyl-1-pyrroline in Manuka Honey (Leptospermum scoparium).

Manuka honey from New Zealand is known for its exceptional antibacterial activity, which is due to high amounts of the 1,2-dicarbonyl compound methylglyoxal (MGO). MGO in manuka honey is formed via non-enzymatic dehydration from dihydroxyacetone (DHA) during honey maturation. MGO and DHA are highly reactive substances, leading to a variety of unique chemical reactions. During Strecker reaction between proline and MGO, 2-acetyl-1-pyrroline (2-AP), an important aroma compound, is formed. Using liquid-liquid extraction and gas chromatography-mass spectrometry analysis, 2-AP was identified unambiguously in manuka honey for the first time. Quantitation was carried out via external matrix calibration, using a synthetic 2-AP standard and artificial honey. The 2-AP concentration in 11 commercial samples of manuka honey ranged from 0.08 to 0.45 mg/kg. For manuka honey samples containing MGO in concentrations above 250 mg/kg, significantly higher amounts of 2-AP were found when compared to non-manuka honeys. When high amounts of MGO were artificially added to non-manuka multifloral honey, an increase of the 2-AP concentration from 0.07 to 0.40 mg/kg after 12 weeks of storage at 37 °C was observed, concomitant with a significant increase in the concentration of 5-hydroxymethylfurfural (HMF). No increase of 2-AP was found during storage at ambient temperature. 2-AP together with MGO can be a suitable parameter for the quality control of manuka honey.

Association Rules Optimization using Artificial Bee Colony Algorithm with Mutation

data mining, Association rule mining is one of the popular and simple method to find the frequent item sets from a large dataset. While generating frequent item sets from a large dataset using association rule mining, computer takes too much time. This can be improved by using artificial bee colony algorithm (ABC). The Artificial bee colony algorithm is an optimization algorithm based on the foraging behavior of artificial honey bees. In this paper, artificial bee colony algorithm with mutation operator is used to generate high quality association rules for finding frequent item sets from large data sets. The mutation operator is used after the scout bee phase in this work. In general the rule generated by association rule mining technique do not consider the negative occurrences of attributes in them, but by using artificial bee colony algorithm (ABC) over these rules the system can predict the rules which contains negative attributes. Keywordsbee colony (ABC), Mutation, Association rule, Support, Confidence, Frequent item set, Data mining.