Iranian Honey Research Articles

Comparative study of physiochemical properties in Iranian multi-floral honeys: Local vs. commercial varieties

Iran, a leading honey-producing country, faces challenges in honey exports. This study aimed to evaluate the melissopalynological and physicochemical characteristics of local honeys belonging to Iranian flora, and compare them with Iranian commercial honeys. For this purpose, seven local honey samples were collected from Iran's renowned floristic regions, alongside seven commercial multi-floral honeys from a supermarket. Moisture content (MC), total solids (TS), pH, free acidity (FA), ash, electrical conductivity (EC), sugar profile, hydroxymethyl furfural (HMF), diastase number (DN), and proline were assessed. The sugar profile was analyzed by high-performance liquid chromatography with a refractive index detector (HPLC-RID). Pollen analysis classified local honeys as multi-floral. The results revealed that MC, TS, pH, FA, ash, and EC values in both local and commercial samples conformed to approved standards (Codex Alimentarius and European Union). Two local and one commercial sample exclusively satisfied the sucrose standard level. Of the local honeys, two samples complied with HMF standard, while five fulfilled DN criterion, and four had proline values within acceptable ranges. Conversely, HMF (56.32–228.11 mg/kg), DN (3.13–7.22 Schade units/g), and proline (109.84–173.86 mg/kg) levels in all commercial samples failed to meet the standard. A significant correlation was found between ash and EC (r = 0.915, p < 0.01) in local honeys, whereas no strong correlation (r = 0.299) existed in commercial samples. Hierarchical cluster analysis confirmed that Iranian honeys lacked clustering by botanical origin, possibly due to extensive sugar adulteration or thermal treatment. Overall, study findings confirmed the significantly inferior quality of Iranian commercial honeys compared to local varieties, albeit some local samples also exhibited quality concerns. Accordingly, it is recommended that regulatory bodies provide periodic training for beekeepers and establish monitoring programs to enhance honey quality, thereby boosting Iran's share in the global honey export market.

Sensitive Electrochemical Sensor for Gallic Acid Determination in Honey Based on g-C3N4/AuNPs Modified Carbon Paste Electrode

Gallic acid (GA) is a well-known polyphenol that occurs naturally in plants and is used as a chemical marker or standard antioxidant in analytical research. Here, a carbon paste electrode was modified with a nanocomposite of graphitic carbon nitride and gold nanoparticles (g-C3N4/AuNPs/CPE). The g-C3N4/AuNPs was characterized by Fourier transform infrared spectroscopy, X-ray diffraction, energy-dispersive X-ray spectroscopy, and scanning electron microscopy. Electrochemical impedance spectroscopy, cyclic voltammetry, and differential pulse voltammetry methods were used to investigate the electrochemical behavior of GA on the electrode. EIS analysis exhibited lower charge-transfer resistance in g-C3N4/AuNPs/CPE than CPE; 250 vs 1500 Ω, respectively. The g-C3N4/AuNPs/CPE was used to GA sensing with limit of detection and linear response range of 0.025 and 0.16–4.10 μM, respectively via DPV. Then, the GA content in Iranian honey samples with different floral origins such as Ziziphus, Barberry, Thyme, Astragalus, Eucalyptus and Coriander was successfully determined. According to result, the fabricated electrochemical sensor could be useful for GA evaluation in food samples.

Production of the First Effective Immune Equine Serum Antivenom against Iranian Honey Bees (Apis mellifera meda)

Production of the First Effective Immune Equine Serum Antivenom against Iranian Honey Bees (Apis mellifera meda)

Effect of ultrasound technique to improve quality of Iranian industrial honey by controlling crystallization process.

This experiment aimed to assess the effects of ultrasound techniques on the quality of Iranian industrial honey. Honey samples were subjected to ultrasound waves at different frequencies and various parameters. The results showed that both ultrasound treatments (30 or 42 kHz) changed the physical, biochemical, antioxidant, and antibacterial characteristics of honey. Ultrasound treatments at 20 or 45°C for 1, 5, or 10 min reduced moisture, acidity, sugars, ABTS levels, 5-hydroxymethylfurfural content, clostridium, aerobic mesophilic bacteria count, and osmophile count while increasing diastase, phenol, and proline levels. Ultrasound treatment of honey samples at 30 and 42 kHz and different temperatures for varying durations led to a decrease in acidity after 90 and 180 days. Treating honey samples with 42 kHz ultrasound at 45°C for 10 min led to a significant reduction in the amount of reducing sugar. Ultrasonication at different frequencies and temperatures led to higher levels of phenol, ABTS, and proline production, along with a considerable decrease in the total count of aerobic mesophilic bacteria. Our study unveils the potential of ultrasonication to enhance honey quality through multifaceted improvements. Treatment significantly augmented phenolic content and antioxidant capacity, opening avenues for novel honey preservation and quality enhancement strategies. Additionally, ultrasonication effectively controlled honey crystallization while simultaneously improving biochemical, antioxidant, and antibacterial properties. This demonstrates its potential as a comprehensive strategy for honey quality improvement.

A paper-based optical tongue for characterization of iranian honey: Identification of geographical/botanical origins and adulteration detection

A paper-based optical tongue for characterization of iranian honey: Identification of geographical/botanical origins and adulteration detection

Analysis of Elements and Physicochemical and Microbial Properties of Iranian Honeys.

Honey is one of the most valuable food products, which, in addition to its nutritional value, also has therapeutic properties. In our study, the physicochemical (Brix, viscosity, free acid content, pH, moisture, diastase activity, 5-hydroxymethylfurfural (HMF), proline content, sugars content, and reducing sugars content) and microbial (mold and yeast content) characteristics and 15 element contents (As, Cd, K, Al, Pb, Hg, Ba, Ni, Na, Ca, Mg, Fe, Mn, Zn, and Se) of the samples were evaluated. Among the essential elements, the maximum mean was related to K (630 ± 50.8mg/kg), and the minimum mean was related to Se that was lower than the limit of detection. Also, among all toxic elements, the maximum mean was related to Ni (234 ± 54.7µg/kg), and the minimum mean was related to Hg that was lower than the limit of detection. Furthermore, the mean of free acidity, pH, °Brix, moisture, diastase content, HMF, and proline content was 35.4 ± 1.27meq/kg, 4.61 ± 0.21, 82.2 ± 3.08, 16.3 ± 0.33%, 9.10 ± 1.14 DN, 21.1 ± 2.65mg/kg, and 482 ± 18.1mg/kg, respectively. Also, the mean percentage of fructose, glucose, and sucrose was 32.4 ± 1.07% (27.5-40.0%), 27.2 ± 0.85% (23.5-31.7%), and 2.28 ± 0.70% (0.72-4.11%), respectively. Finally, the mean of mold and yeast in all samples was 14.2 ± 0.37CFU/g. Also, the principal component analysis and heat map allowed us to determine a more accurate distinction between the physicochemical characteristics of bee honey. The results of our findings showed that in most cases, the results obtained were within the standard range, which indicates the good quality of Iranian honeys.

Effect of Ultrasound on the Physical, Biochemical, Anti-oxidant, and Antimicrobial Properties of Industrial Iranian Honey

Honey is a sweet and nutritious product of Apis mellifera bees. It contains glucose, fructose, water, minerals, vitamins, organic acids, phenolic compounds, and enzymes . Honey has various functional properties such as antioxidant, antibacterial, antifungal, prebiotic, and anticancer, which are essential for human health [4]. Because of its unique properties, honey is known not only as a nutritious spice but also as a medicine with healing properties. Honey is a high-dense sugar solution; it can produce coarse crystals by changing the conditions (such as glucose level, humidity, temperature, impurity, and ecology) and lead to quality loss . This study aimed to determine the effect of ultrasound at two different frequencies of 30 and 42 kHz on the physical, biochemical, anti-oxidant, and antimicrobial properties of industrial Iranian honey. All samples (except control) were treated using ultrasound at 30 or 42 kHz for 1, 5, or 10 minutes at 20 or 45 °C. Then the following parameters were measured: HMF content, pH, acidity, proline, total count of aerobic mesophilic bacteria, diastase, moisture, sucrose, fructose, glucose, the ratio of fructose to glucose, ABTS, osmophiles, phenol, reducing sugars and total sugars on days 1, 30, 90, and 180. In both treatment groups exposed to ultrasound, there were changes in these parameters. The group treated with 42 kHz ultrasound showed a decrease in moisture, pH, acidity, fructose-to-glucose ratio, total sugars, clostridium, total microbial count, mold, osmophiles, HMF, and an increase in diastase, proline, and phenol. The changes were more significant on days 90 and 180. Ultrasound at 42 kHz for 10 minutes at 45 °C could lead to an increase in the quality and shelf life of industrial honey. In conclusion, this technique can be used in the food industry to provide a higher quality product as well as a more successful export.

Genetic Characterizations of the Iranian Honey Bee (Apis mellifera meda Skorikov 1929) Populations Using the Microsatellite DNA Markers.

The genetic characterization of the Iranian honey bee was investigated by analyzing 10 polymorphic DNA microsatellite loci in 300 honey bee samples representative of twenty Iranian provinces. This study evaluated the heterozygosity (Ho and He), the Shannon index, the number of observed alleles, and F-statistics among tested populations as genetic parameters. Our finding demonstrated that the Iranian honey bee populations were described by low genetic diversity in terms of the number of observed alleles, Shannon index, and Heterozygosity values. Most populations had significant deviations from Hardy Weinberg equilibrium cause of heterozygote shortage. Low FST and FIS values proposed the absence or very low genetic diversity within and among A. m. meda populations in the present study. The cluster analysis has categorized the honey bee samples gathered from various regions of Iran into two main groups, including honey bees in the North-West (i.e., North, Northwest, and West) provinces and honey bees in the East-South (i.e., Eastern North, Central part, and Southern) provinces of Iran. Our results also revealed lower genetic differentiation and heterozygosity among tested honey bee populations. The results from this study are consistent with previous investigations in Iran, alarming the loss of genetic diversity in the Iranian honey bee populations, which leads to more homozygosity. This study presented new data and reports on genetic structure in investigated native Iranian honey bee populations, and it will benefit future studies on selection, native biodiversity preservation and other conservation breeding projects.

The effect of fermented pollen supplements on the percentage of cell acceptance rate and royal jelly production in Iranian honey bees (Apis mellifera meda)

Royal jelly (RJ) is a yellowish-white substance secreted by the secretory glands of young worker bees to feed the larvae and the queen throughout its life. This study investigates the effect of fermented pollen supplementation on RJ production and cell acceptance rate in Iranian honey bees. The experiment was conducted in Marivan, Iran, for 30 days in the summer of 2020. Experimental treatments include (1) control (sugar syrup); (2) pollen cake (raw pollen, powdered sugar, and honey) + sugar syrup; (3) unfermented pollen supplement + sugar syrup; and (4) fermented pollen + sugar syrup. RJ production and cell acceptance rate were measured and estimated at different time intervals. The collected data were analyzed in a completely randomized design using SAS9.4 software using the GLM procedure. The results showed that different experimental treatments had a significant effect (p ≤ 0.05) on the amount of RJ produced. Among various treatments, the highest RJ production was related to the fourth treatment (12.53 g), and the control treatment (8.07 g) had the lowest production rate. In addition, the highest mean percentage of cell acceptance rate (97.58%) was related to the fermented pollen + sugar syrup treatment. The results showed a positive and high correlation between the two traits of cell acceptance rate and RJ production. In general, a fermented supplementary diet is more efficient than unfermented rations with the same ingredients, therefore recommendable for the apiculture industry.

Molecular genetic diversity and population structure of Iranian honey bee (Apis mellifera meda) populations: implications for breeding and conservation

Molecular genetic diversity and population structure of Iranian honey bee (Apis mellifera meda) populations: implications for breeding and conservation

RETRACTED: Iranian thyme honey plays behavioral, cellular and molecular important roles as an amazing preventive and therapeutic agent in the brain of Alzheimer's rat model

Alzheimer’s disease (AD) is a progressive neurodegenerative disorder with neuronal loss in the hippocampus. Our aim was to evaluate the effects of Iranian thyme honey (single dose: 2 gr/kg) vs rivastigmine (0.3 mg/kg) in vivo on spatial memory and in vitro on important parameters of oxidative stress as well as quantitative and qualitative studies of hippocampal neurons of AD rat models with this design that 30 days after oral administration of 17 mg/kg AlCl3, 20 AD rats were received that underwent a 6-weeks therapeutic period and another 20 rats underwent a 6-weeks preventive period and also 20 rats were as controls. Y-Maze test was performed to show memory deficiency as well as TBARS and FRAP assays to measure malondialdehyde (MDA) and total antioxidant, respectively. In addition, H&E staining was also done for cell counting and morphological changes. We observed that AD rats with hippocampal damage had more significant errors during the Y-maze test than the control and other rats. Likewise, MDA and neurodegeneration increased in the AD group while in all preventive and therapeutic group's especially Iranian thyme honey, they decreased and conversely, total antioxidant and number of normal cells elevated and healthy neurons were observed in all parts of the hippocampus and cortex. Our results despite the limitations showed the powerful antioxidant properties and cytoprotective effects of Iranian thyme honey vs rivastigmine on hippocampal neurons that consequently enhanced memory and if advanced diagnostic tests in human clinical patients show other more pronounced effects, we have certainly started a key and targeted strategy.

Quality Evaluation of Iranian Honey Collected from Khorasan Province, Iran.

Honey is a prominent nutritional and medicinal production of honey bees, originating from the nectar of flowers. The physicochemical properties of honey serve as indicators of its freshness and originality. The current survey aimed to assess the parameters of quality control, including hydroxymethylfurfuraldehyde [HMF], reducing sugars, fructose/glucose, sucrose, proline content, distaste activity, and free acidity, in 25 honey samples of different brands available in Khorasan Province, Iran. We used the methods suggested by the Association of Official Analytical Collaboration (AOAC, 1995), the International Honey Commission (IHC, 2009), and the Codex Alimentarius Honey Standards for the study. Statistical analysis was performed in Microsoft Excel. The obtained data indicated that eight out of 25 analyzed samples (32%) complied with all the requirements and were generally of acceptable quality. Meanwhile, 17 samples (68%) were unconfirmed by the Iranian Standard Organization (ISO), including 12 samples with a low level of diastase (<8 Schade) and high levels of HMF (>15 mg/kg), two samples with high sucrose levels, two samples with high proline, and one sample with high HMF. These findings suggested their inappropriate storage (time/temperature), heat treatment, and/or adulteration with industrial sugar. According to the results, the examined honey samples produced in Khorasan Province were not of acceptable quality, which highlights the importance of an effective regulatory framework to be evaluated and rectified periodically and accurately to maintain consumer rights, as well as public health.

Effect of Honey Bee Venom on Experimental Autoimmune Encephalomyelitis (EAE) as a Model for Multiple Sclerosis (MS).

Experimental autoimmune encephalomyelitis (EAE) has been widely employed as a model to study multiple sclerosis (MS). Interleukin-27 (IL-27) inhibits Th17 activity and breaks the normal activity of effector T cells which cause autoimmunity. Bee venom (BV) has been used as a form of medicine from the time of ancient Greece and China. BV and BV-derived active components might have potent therapeutic effects on refractory immunological and neurodegenerative diseases, such as MS. This study aimed to investigate the effect of Iranian honey bee venom on the progression of EAE in mice. Initially, EAE was induced in 12 female C57BL/6 mice through immunization with an emulsion of myelin oligodendrocyte glycoprotein 35-55 (MOG35-55) in Complete Freund's adjuvant (CFA), followed by administration of pertussis toxin (PTx) in phosphate buffer. Following the appearance of clinical signs, the mice were treated intraperitoneally with BV. Histopathological and immunological studies were investigated, and EAE was induced in animals within 9-14 days. Results revealed a significant reduction in IL-27 levels following EAE induction in mice. However, BV-treated mice showed a significant increase in IL-27, compared to controls. Histopathology results revealed that the number of inflammatory cells was reduced in the brain parenchyma following BV treatment. Based on the results obtained in the present study, BV may be a suitable candidate for the treatment of inflammatory diseases, such as MS.

Vairimorpha ceranae was the only detected microsporidian species from Iranian honey bee colonies: a molecular and phylogenetic study.

Nosemosis caused by Vairimorpha ceranae is one of the most important threats to honeybee colonies worldwide. This study aimed to determine the prevalence and intensity of Vairimorpha infection in different types of colonies and locations in Iran. In October 2017 and May 2018, 376 colonies from 97 apiaries were selected for each month according to a randomly clustered design. By considering 3-5 colonies for each apiary, 20 adult bees as pooled samples were collected from each colony. In microscopic analysis, 46.52% and 46.1% of samples in May and October showed Vairimorpha spores, respectively. The infection intensities in May and October were 5.94 ± 0.19 (× 106) and 5.86 ± 0.23 (× 106) spores/bee in a pooled sample, respectively. The mean infection intensity ranged from 1.8 to 12.5 (× 106) spores/bee. Statistically, there were no significant differences in the prevalence and intensity of V. ceranae infection between May and October samples. No significant differences were found among the prevalence rates of infection in the types of colonies; however, the intensity was significantly higher in migratory and mountainous colonies in May and only in migratory colonies in October. There was a significant correlation between the prevalence and intensity of V. ceranae infection (r2 = 0.695). PCR analysis showed that the samples were only infected with V. ceranae. No intraspecific variation to V. ceranae was found by direct sequencing of the amplified fragment of 16S rRNA. The obtained sequence was mainly 100% similar to those of V. ceranae isolates from European countries.

Investigation of protein content and the relationship between enzymatic activity and antimicrobial effect of Iranian honey samples

Investigation of protein content and the relationship between enzymatic activity and antimicrobial effect of Iranian honey samples

Examination the antioxidant potentials and antidiabetic properties of phenolic extracts of some Iranian honeys

Introduction: In most ancient cultures, honey has been used for both nutritional and medical purposes. Objectives: In this research, phenolic extracts of four Iranian honeys were evaluated to determine the antioxidant potentials using DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging, nitric oxide (NO) radical scavenging and reducing power by ferric reducing activity of plasma (FRAP) method. Additionally, anti-diabetic properties of honey and phenolic extracts were evaluated by determination of α-amylase and α-glucosidase inhibition. Patients and Methods: Besides, reducing potential was evaluated by ferric-reducing antioxidant power method. Moreover, determination of phenolic and flavonoid contents was performed. Moreover, inhibition of α-amylase and α-glucosidase of honey and phenolic extracts were evaluated. Results: With considering to antioxidant potentials, Gavan (Astragalus) sample showed the greatest phenolic (3817±1.52 mg GAE/100 g), flavonoid contents (3.1±0.005 mg QE/100 g), and DPPH radical scavenging (IC50 = 2±0.003 mg/mL). Bahareh honey had the highest NO radical scavenging (IC50=0.0403±0.0009 mg/mL) and Meymand honey possessed the highest reducing potential by FRAP method (IC50=0.0018±0.000003 mg/mL). The maximum inhibition of α-glucosidase was shown in Meymand honey extract (46±0.1%). After sugar isolation, Zataria honey had the highest inhibition of α-glucosidase (54±0.6%) and the mode of α-amylase inhibition was noncompetitive by this honey. Whole extract (23±0.1%) and phenolic extract of Gavan honey presented the maximum inhibition of α-amylase (31.2±0.1%). Conclusion: Honey samples showed antioxidant potentials and anti-diabetic properties by retardation of α-amylase and α-glucosidase.

Evaluating the quality of commercial Iranian honeys

Evaluating the quality of commercial Iranian honeys

The sources and quality of Iranian honey

Iran is one of the largest honey-producing countries worldwide and is considered as an important source of honey for international markets. However, since Iran is not registered for honey export to Europe, the quality of Iranian honey remains unknown to European traders. As the first step in filling this gap, we analyzed 225 honey samples using palynology, sensory, nuclear magnetic resonance (NMR) and conventional physicochemical analyses as outlined by the European Union coordinated control plan. The results show that while various types of genuine unifloral honey can be harvested in Iran, 85% of collected samples were adulterated. Performing principal component analysis on physicochemical parameters reveals that feeding tablet sugar and syrup of C4 origin to bees during the foraging season is a common mode of fraud. Replacement of natural nectar with sugar syrup together with presence of intensive aftertaste from Taraxacum and Eryngium affect the taste of unifloral honeys produced in Iran.

Antibacterial activity and mechanism of action of some Iranian honeys compared to manuka honey against multidrug-resistant respiratory and urinary infections

Increasing antibiotic resistance among bacterial pathogens requires the identification of alternative therapeutic agents, especially the natural ones. The importance of honey in health care was document from the oldest medical literature. Currently, the antimicrobial properties of honey, especially for the prevention of resistant strains, has been considered. However, from mechanistic insights, this activity has received less attention. In this study, the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of 6 Iranian honey samples in comparison with Manuka honey against reference strains of Staphylococcus aureus ATCC 29737, Pseudomonas aeruginosa ATCC 27853, Klebsiella pneumonia ATCC 10031, Escherichia coli ATCC 10536 and 22 multidrug-resistant clinical strains of respiratory and urinary infections were determined. The survival of bacteria and their ability to produce biofilm in the presence of a bactericidal concentration of honey were monitored. The effect of the most powerful honey and Manuka on the structure of reference strains of the Staphylococcus aureus and Pseudomonas aeruginosa was investigated using Field Emission Scanning Electron Microscopy (FESEM) and flow cytometry analysis. Exposure of bacteria to the studied honey generally resulted in high antimicrobial and anti-biofilm activity of Iranian honey compared to Manuka honey. Using SEM, division inhibition accompanied by loss of structure and marked changes in cell shape and surface were observed in the presence of honey. Analysis of flow cytometric charts revealed synchronous increasing and decreasing in the size of honey-treated strains. Present results indicated that Iranian honey samples possess the potential to be an effective inhibitor of multi-drug resistant strains.

Bacteriocin activity of various iranian honey-associated bacteria and development of a simple medium for enhanced bacteriocin activity.

Honey is a promising source of bacterial strains producing metabolites with antimicrobial activity. There is a great variety in the antimicrobial activity of honey from different areas of nature. Therefore, the aim of present study was to investigate the antibacterial activity of Iranian honey from different regions and to optimize the culture condition for the highly potent bacterial isolate. Honey samples were collected from ten different regions of Iran and were screened for bacteriocin-producing bacteria. The best bacteriocin-producing strain was characterized and identified by 16S rDNA analysis. One-factor-at-a-time method was used for optimization of culture medium and the yield and time-course of bacteriocin production were compared in both shake flask and bio-reactor. The Bacillus subtilis SB1 that was isolated from Sabalan honey showed potent antibacterial activity with prominent thermal stability. The optimum medium for the bacteiocin production was a yeast extract-based medium. The optimum incubation temperature for bacteriocin production was 34°C. Bacteriocin production was higher near neutral pH conditions than that produced at acidic or alkaline environment. The results of cell growth and bacteriocin assays revealed that the exponential phase of growth and antibacterial compounds production was started rapidly in bioreactor than flask. Findings of this study supported the folkloric application of honey against some infectious diseases. B.subtilis SB1 that isolated from Sabalan honey was a potential source for bacteriocins-like compounds. Our studies suggested a simple buffered nitrogen-based medium for SB1 growth and bacteriocin activity in both shake flask and bioreactor.