Iranian Honey Research Articles

Heavy metals determination in honey samples using inductively coupled plasma-optical emission spectrometry.

BackgroundHoney contains a complex mixture of carbohydrates and other minor substances. Elements are minor constituents of honey that may threaten the human health in excess concentrations. So, determining the metals in honey helps its quality control as a food product. The aim of this study was to determine the concentrations of some metals in Iranian honey.MethodsThis study was performed in four regions of Ardabil, a province of Iran. Honey samples (n = 25) were digested in microwave oven by nitric acid and hydrogen peroxide, then analyzed using inductively coupled plasma– optic emission spectrophotometry (ICP-OES).ResultsNo significant differences were observed in cadmium, zinc, nickel, and chromium levels between regions (P > 0.05). Zinc was the most abundant metal in honey samples (1481.64 μg/kg). Some metals had higher concentrations in the East region because of existence more industries there. The highest mean of lead level was 935.48 μg/kg in the East and the lowest was 205.4 μg/kg in the South region. The concentrations of metals were compared with recommended limits for foods. Some of them were higher than standard levels (lead) and some were lower than those (cadmium).ConclusionsMetals are released into the environment through their use in industrial processes and enter the food chain from uptake by plants from contaminated soil or water. Metals concentration in various places depends on many variables, leading to their different concentrations in honey. Some control measures like the quality control of food products, monitoring the soil in agricultural regions and limiting the use of fertilizers are recommended.

In vitro characterization of antibacterial potential of Iranian honey samples against wound bacteria

Incidence of multidrug-resistant pathogens is increasing, thus discovery of alternative therapeutic agents is needed. Honey possesses therapeutic potential, including wound-healing properties and antimicrobial activities. Although antimicrobial activity of honey has been effectively established, the antibacterial properties of Iranian honey samples on wound microorganisms have not been conducted and compared with honey gold standards. Using standard methods, the antibacterial activities of 53 Iranian honey samples and two laboratory-synthesized honeys against 16 drug-resistant clinical isolates and four reference strains were determined. Different concentrations of honey were tested against each type of microorganism to determine the minimum inhibitory concentration (MIC) or minimum bactericidal concentration. The selected honeys with high levels of phenolic equivalent were studied for kinetics of bacterial killing. Also, the ability of selected honey samples to prevent biofilm formation was checked by crystal violet assay. The MICs of Iranian honey samples ranged from 3.12 to >50 % (w/v), and their efficacies were also determined. Nine samples with the highest antibacterial activities were selected. Survival of Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa was reduced significantly by selected honeys. Moreover, all selected honey samples were able to prevent biofilm formation of S. aureus and P. aeruginosa strains at concentrations of ≤25 %. The results of the present study have shown that selected Iranian honey samples had lower MIC values than reported standard honeys such as Manuka and Tualang and could be potentially used as alternative medical grade honey, particularly against S. aureus, P. aeruginosa, and E. coli.

Anti-HIV-1 activity of eight monofloral Iranian honey types.

Monofloral Iranian honeys from eight floral sources were analyzed to determine their anti-HIV-1 activities as well as their effects on lymphocyte proliferation. The Peripheral Blood Mononuclear Cells (PBMCs) used in this study were prepared from five healthy volunteers who were seronegative for HIV, HCV, HBV and TB. The anti-HIV-1 activity of eight different honeys was performed by quantitative polymerase chain reaction (PCR) assay and high pure viral nucleic acid kit. The results demonstrated that monofloral honeys from Petro selinum sativum, Nigella sativa, Citrus sinensis, Zataria multiflora, Citrus aurantium and Zizyphus mauritiana flowers had potent anti-HIV-1 activity with half maximal effective concentration (EC50) values of 37.5, 88, 70, 88, 105 and 5 µg/ml respectively. However, monofloral Iranian honeys from Astragalus gummifer and Chamaemelum nobile flowers had weak anti-HIV-1 activity. The frequency and intensity of CD4 expression on PBMCs increased in the presence of all honey types. CD19 marker were also increased after the treatment with monofloral honeys from Z.multiflora and N. sativa. The anti-HIV-1 agent in monofloral honeys from P.sativum, N. sativa, Z. multiflora and Z. mauritiana flowers was detected by spectroscopic analysis as methylglyoxal. Time of drug addition studies demonstrated that the inhibitory effect of methylglyoxal is higher on the late stage of HIV-1 infection. The result demonstrated that methylglyoxal isolated from monofloral honey types is a good candidate for preclinical evaluation of anti-HIV-1 therapies.

Microsatellite genetic diversity of Apis mellifera meda skorikov.

The genetic diversity of three Iranian honey bee populations (Apis mellifera meda) was studied using morphological and microsatellite loci in south Iran. For this purpose ten morphological characters and five microsatellite loci were studied. Morphometric analysis resulted in a distinct classification of three investigated populations but showed low diversity among them. The grouping results of the diversity study by microsatellite markers were in agreement with the results of morphometry. The cluster analysis showed that the honey bees have clustered together in one group. These populations displayed low variability estimated from both the number of alleles and heterozygosity values. Genetic differentiation within the populations is low and low heterozygosity was also presented between diverse populations. These results indicate the existence of a single population structure. The results of current research confirmed us the previous findings concerning morphological and biochemical indications of uniformity in the honey bee population of the south Iran in spite of the fact that the cities which was studied by us separated from each other by a distance of 500 km.

Antibiotic Residues in Iranian Honey by Elisa

The aim of this study was to determine antibiotic residues in honey collected from northern part of Iran. The presence of antibiotic residues in honey may be harmful to human health. In this study, the enzyme linked immune assay method was used to investigate of antibiotic residues in honey samples (n = 135) collected randomly from Qazvin Province, Iran. The analyzed antibiotics included enrofloxacin, penicilin, chloramphenicol, gentamicin, tylosin, tetracycline, and sulfonamide. Result show that the range of antibiotic radiuses value was 0.0–72.1 ng/g, besides; the highest percentage of antibiotic residues in honey samples was the enrofloxacin (20.7%). The highest and lowest mean contamination (ng/g) belonged to enrofloxacin (10.8 ± 1.6) followed by Penicillin (4.4 ± 2.9), and chloramphenicol (0.1 ± 0.1). The highest level of antibiotic residues (71.85%) was seen in honey samples collected during the autumn season (P < 0.05).

The Effects of Age of Grafted Larvae and of Supplemental Feeding on Performance of Iranian Honey Bee Colonies (Apis Mellifera Meda)

Abstract The performance of bee colonies greatly depends on the quality of the queens. The current research was conducted at the apiary of the Faculty of Agriculture, Zanjan University, in Zanjan, Iran. Together, 24 rearing colonies were assigned to 4 grafting larvae age groups: 1-day-old, 2-day-old, 3-day-old, and emergency queens. Two feeding groups, fed and not fed, were created. The effects of reared queens on biological characteristics and performance of honeybee colonies (Apis mellifera meda) headed by those queens were measured. Age of grafted larvae significantly influenced the results. The performance ratios of the most efficient colonies headed by queens reared from 1-day-old larvae compared with the least-efficient queens reared from 3-day-old larvae were 118% in brood production, 140% in bee population, and 154% in honey production. However, the age of grafted larvae did not affect colony defense behavior. Supplemental feeding of rearing colonies increased brood production to 111%, bee population to 116%, and honey production to 115%. A combination of the effect of age of larvae and supplemental feeding resulted in twice as much honey (12 kg) produced by colonies with queens reared from 1-day-old larvae in fed rearing colonies compared to those with queens raised from 3-day-old larvae in unfed rearing colonies.

Effect of honey as an immunomodulator against invasive aspergillosis in BALB/c mice

SummaryInvasive aspergillosis has become an increasing problem in immunocompromised patients in recent years. Due to increased antimicrobial resistance, natural agents with medicinal and immunomodulatory effects have gained more attention. Honey is a natural substance with documented antimicrobial, anti-inflammatory and wound healing effects. In this study the effects of three Iranian honeys on some aspects of innate immunity and survival rate during invasive aspergillosis were investigated. For this purpose, mice were divided into 10 groups (honey alone, honey and infection, negative and positive controls) each containing 10 individuals. Mice were treated with honey (1.5g/kg BW/orally) for 10 days. At day 6, Aspergillus fumigatus conidia (5 × 105/ml) were injected intravenously to the infected groups. Mice were euthanized at day 11, and spleen and peritoneal cell culture was performed. Pro-inflammatory cytokine production using ELISA and killing assay was also performed. For survival rate, 10 mice from each infected group were considered and monitored for 30 days. The results showed that honey treatment could significantly increase IL6 and IL1β production in infected mice as well as improve macrophage killing (p < 0.05). Also mice treated with honey had a greater survival time than the infected group. Our results suggest that honey could boost the immune system and increase survival time in invasive aspergillosis.

Antifungal Activity of Iranian Honeybees Against C andida, A spergillus Species and T richophyton R ubrum

Due to limited available antifungal agents, searching for new antifungal agents stimulated research on new chemotherapeutic agents in natural products. The current study was carried out to assess the antifungal activity of different Iranian honeys with different botanical origin against some important fungi, such as azoles-resistant Candida species. Seven types of honey samples including Thymus vulgaris, Alfalfa, Citrus, Zizyphus, Astragalus, Chamaemelum nobile and multiflora honey were collected. Minimum inhibitory concentrations microdilution broth method was used for assessing antifungal activity of honeys. The results revealed variations in the antifungal activity of the examined honey samples. Alfalfa and Thymus honeys were significantly more effective than other honeys and Zizyphus and Chamaemelum honeys exhibited lowest antifungal activity. Trichophyton rubrum and C. krusei had the highest and lowest sensitivity to the honey's antifungal activity among fungal species, respectively. No statistically significant differences in susceptibility to honey samples were determined among fluconazole resistance and susceptible Candida species. Practical Applications In the present study, the antifungal properties of honeys with different botanical origin against pathogenic fungal species were examined. Furthermore, the inhibitory properties of the honeys against fungal strains resistant to existing antifungal drugs were assessed. Antimicrobial properties of plants which were the origin of honeys in this study were also investigated. Results showed a direct correlation between the type of nectar for honeys and antifungal properties. More importantly, inhibitory effects of honeys against several fungal species and Trichophyton rubrum, particularly resistant Candida species was shown. It seems that appropriate concentrations of honey can be used for controlling and treating of dermatophytosis and dermatomycosis.

A Study of Polymorphisms in Three Loci Known to Influence Defensive Behavior Using PCR-SSCP and Direct Sequencing in the Iranian Honey Bee Population

Polymerase Chain Reaction (PCR) and subsequent separation of PCR products on polyacrylamide gels to find Single Stranded Conformation Polymorphisms (SSCP) was used for three loci, known from studies in North America to affect defensive behavior (Quantitative Trait Loci (QTL) sting1–3), on samples of Iranian honey bee (Apis mellifera L.). In the present study these loci were amplified with specific primers for sting1–3. The analysis of these loci using SSCP, created different patterns among samples. The polymorphisms observed in this study of the Iranian population of honey bees are a first step toward the eventual implementation of a genetic marker-based selection program to reduce defensive behavior. Now, because of this study, the markers are available to conduct more research so that the association of these polymorphic loci and aggressive traits in the Iranian honey bee population can be determined.

Regular dorsal dimples and damaged mites of Varroa destructor in some Iranian honey bees (Apis mellifera)

The frequency of damaged Varroa destructor Anderson and Trueman (Mesostigmata: Varroidae) found on the bottom board of hives of the honey bee, Apis mellifera L. (Hymenoptera: Apidae) has been used as an indicator of the degree of tolerance or resistance of honey bee colonies against mites. However, it is not clear that this measure is adequate. These injuries should be separated from regular dorsal dimples that have a developmental origin. To investigate damage to Varroa mites and regular dorsal dimples, 32 honey bee (A. mellifera) colonies were selected from four Iranian provinces: Isfahan, Markazi, Qazvin, and Tehran. These colonies were part of the National Honey bee Breeding Program that resulted in province-specific races. In April, Varroa mites were collected from heavily infested colonies and used to infest the 32 experimental colonies. In August, 20 of these colonies were selected (five colonies from each province). Adult bees from these colonies were placed in cages and after introducing mites, damaged mites were collected from each cage every day. The average percentage of injured mites ranged from 0.6 to 3.0% in four provinces. The results did not show any statistical differences between the colonies within provinces for injuries to mites, but there were some differences among province-specific lines. Two kinds of injuries to the mites were observed: injuries to legs and pedipalps, and injuries to other parts of the body. There were also some regular dorsal dimples on dorsal idiosoma of the mites that were placed in categories separate from mites damaged by bees. This type of classification helps identifying damage to mites and comparing them with developmental origin symptoms, and may provide criteria for selecting bees tolerant or resistant to this mite.

Mitochondrial DNA Variation in the CoxI–CoxII Intergenic Region among Turkish and Iranian Honey Bees (Apis mellifera L.)

The western honey bee, Apis mellifera L., is native to Africa, the Near and Middle East, and Europe. Based on morphometrics, the Near Eastern subspecies, Anatolian (A. m. anatoliaca), Caucasian (A. m. caucasica), and Iranian (A. m. meda), were grouped within the Middle Eastern O branch (Ruttner 1988; Kauhausen-Keller et al. 1997); however, mtDNA analysis has shown that they belong to the East Mediterranean C lineage (Smith et al. 1997; Palmer et al. 2000; Franck et al. 2000, 2001; Kandemir et al. 2006; Ozdil et al. 2009). According to Ruttner’s (1988) morphometric analysis, A. m. anatoliaca occupies nearly all of Turkey, A. m. caucasica is found in the northeastern part of Turkey, and A. m. meda is found primarily in Iran but also in the southeastern part of Turkey. Length and restriction site polymorphisms within the CoxI–CoxII intergenic region of the honey bee mitochondrial genome have been particularly useful in differentiating evolutionary lineages and groups of subspecies (Cornuet and Garnery 1991; Hall and Smith 1991; Garnery et al. 1992, 1993; Franck et al. 2000, 2001; Palmer et al. 2000). For example, DraI restriction of this region revealed more than 50 haplotypes in lineages A and M (Garnery et al. 1992, 1993; Franck et al. 2000, 2001) but only six haplotypes in lineage C (Franck et al. 2000; Kandemir et al. 2006; Ozdil et al. 2009), including one discovered recently (Ozdil et al. 2009).

Determination of Flavonoid Markers in Honey with SPE and LC using Experimental Design

Determination of flavonoid markers quercetin, hesperetin, and chrysin, found in north Iranian citrus honey samples, was carried out by solid phase extraction (SPE) and isocratic liquid chromatographic separation using central composite design. Optimum conditions for SPE were achieved using 10 mL methanol/water (13:87, v/v, pH = 7) as the washing solvent and 4 mL methanol for elution. Good clean-up and high recovery >90% were observed for all analytes. The use of water/ACN/THF/AcOH (54:36:5:5, v/v) was found to serve as the optimum mobile phase composition and allowed for the separation of analytes from endogenous compounds present in honey. SPE parameters, such as maximum loading capacity and breakthrough volume, were also determined for each analyte. Limit of detection, linear range, recovery, repeatability of retention times, and peak heights were 3.11 × 10−8–4.44 × 10−8 g g−1, 0.50–50.0 μg mL−1 (R 2 > 0.99), 90.7–96.9%, 3.0–3.6%, and 1.0–2.6%, respectively. Precision of the overall analytical procedure, estimated by five replicate measurements for quercetin, hesperetin and chrysin in citrus honey, as well as the relative standard deviations were 4.3%, 3.8%, and 5.5%, respectively.

Morphometric and genetic variability of honey bee (Apis mellifera L.) populations from northern Iran

SummarySamples from seven honey bee populations; five Apis mellifera meda from Iran and two from Turkey, one belonging to A. m. meda and one to A. m. caucasica, were investigated using morphometric, mtDNA, and microsatellite analyses. Morphometric analysis revealed three distinct clusters. The first included all Iranian populations, whilst the second and third contained the meda and caucasica bees from Turkey. No variation was observed in the DraI restriction of COI-COII intergenic region in mitochondrial DNA, yielding a single C lineage mitotype. The analysis of microsatellite loci, A7, A24, A28, A43 and A113 revealed, a considerable level of genetic diversity, with average expected heterozygosity ranging from 0.596±0.188 to 0.695±0.174 and average number of alleles per locus from 4.4 to 8.0 among Iranian honey bee populations.

Fungicidal potential of different Iranian honeys against some pathogenic Candida species.

Fungicidal potential of different Iranian honeys against some pathogenic Candida species.

Fungicidal potential of different Iranian honeys against some pathogenic Candida species

SummaryHoney is well known to possess a broad spectrum of activity against medically important organisms. The purpose of this study was to investigate the anti-candidal activity of 28 locally produced honeys from two floral sources, 14 from southern and 14 from northern Iran, against some pathogenic Candida species such as Candida albicans, C. parapsilosis, C. tropicalis, C. kefyr, C. glabrata, and C. dubliniensis. A microdilution broth method was used to assess the activity of the selected honeys against six Candida species. The honeys were tested at 40 concentrations ranging from 20 to 60 % (v/v). Both northern and southern honeys possessed in vitro antifungal activity against the six Candida species tested. Little or no anti-candidal activity was seen at concentrations lower than 24%. Increased honey concentrations resulted in reduced growth of Candida species. The highest inhibitory effect of different honeys was demonstrated in C. tropicalis, C. glabrata and C. dubliniensis. Candida kefyr and C. albicans showed more resistance to fungicidal activity of honeys, whereas C. glabrata was least affected. All honeys were able to produce complete inhibition of candidal growth with minimum fungicidal concentrations ranging from 29 to 56 %. Overall, varying sensitivities to the anti-candidal properties of different honeys were observed with six species of Candida emphasizing the variability in the antifungal effect of honey samples. Further research is needed to assess the efficacy of honey as an inhibitor of candidal growth in clinical trials, especially in treatment of patients with candidiasis.

Efficacy of Intra-peritoneal Administration of Iranian Honey in Prevention of Post-operative Peritoneal Adhesions

Efficacy of Intra-peritoneal Administration of Iranian Honey in Prevention of Post-operative Peritoneal Adhesions

Morphometric characterization of Iranian (Apis mellifera meda), Central Anatolian (Apis mellifera anatoliaca) and Caucasian (Apis mellifera caucasica) honey bee populations

SummaryMorphological characters of Apis mellifera meda sampled from Urmia, Tehran and Tabriz in Iran, Apis mellifera anatoliaca from Kirsehir and Beypazari in Central Anatolia, and Apis mellifera caucasica from Posof in Northeast Anatolia were analysed using standard morphometric methods. Sixteen morphometric measurements were made on 20 worker bees each from 5 colonies from 6 localities in Iran and Turkey. Univariate analysis showed that significant differences for all characters were found among honey bee samples from 6 localities (P < 0.05). The honey bees from Iran (Urmia, Tehran and Tabriz) were significantly smaller than the Caucasian honey bee from Posof and Central Anatolian honey bees from Kirsehir and Beypazari in most of the size characters. The Caucasian honey bee from Posof had the highest values in the hair length on tergite 5, forewing length, forewing width and hind leg length which separate it from all other groups. Discriminant analysis showed that the Iranian, Central Anatolian and Caucasian honey bee populations constituted distinct clusters. Discriminant analysis correctly classified 66.7% of the samples to their actual localities. Iranian honey bee samples from 3 different localities appear not to be entirely distinct sub-populations. The samples from Urmia, Tehran and Tabriz overlapped to a large extent, while the samples from Kirsehir, Beypazari and Posof formed apparent non- overlapping clusters. Mahalanobis distances confirmed that Caucasian bees were closer to Central Anatolian bees than to Iranian bees.

High ascorbic acid content in some Iranian honeys. Chemical and biological assay.

A high content of Vitamin C (over 100 mg/100 g) in three samples of honey from a mountainous area (Damavand) in Iran is reported. The vitamin was determined by thin layer chromatography of its 2, 4 dinitro phenyl osazone, as well as by the less specific 2,6 dichlorophenol indophenol method. The effect of one of these samples on the prevention and cure of scurvy in the guinea pig indicates that it contained compounds with vitamin C activity.