Pollen Analysis of Natural Honeys from the Central Region of Shanxi,North China

Based on qualitative and quantitative melissopalynological analyses, 19 Chinese honeys were classified by botanical origin to determine their floral sources. The honey samples were collected during 2010–2011 from the central region of Shanxi Province, North China. A diverse spectrum of 61 pollen types from 37 families was identified. Fourteen samples were classified as unifloral, whereas the remaining samples were multifloral. Bee-favoured families (occurring in more than 50% of the samples) included Caprifoliaceae (found in 10 samples), Laminaceae (10), Brassicaceae (12), Rosaceae (12), Moraceae (13), Rhamnaceae (15), Asteraceae (17), and Fabaceae (19). In the unifloral honeys, the predominant pollen types were Ziziphus jujuba (in 5 samples), Robinia pseudoacacia (3), Vitex negundo var. heterophylla (2), Sophora japonica (1), Ailanthus altissima (1), Asteraceae type (1), and Fabaceae type (1). The absolute pollen count (i.e., the number of pollen grains per 10 g honey sample) suggested that 13 samples belonged to Group I (<20,000 pollen grains), 4 to Group II (20,000–100,000), and 2 to Group III (100,000–500,000). The dominance of unifloral honeys without toxic pollen grains and the low value of the HDE/P ratio (i.e., honey dew elements/pollen grains from nectariferous plants) indicated that the honey samples are of good quality and suitable for human consumption.     

Antibacterial activity of different honeys against pathogenic bacteria

To study the antimicrobial activity of honey, 60 samples of various botanical origin were evaluated for their antimicrobial activities against 16 clinical pathogens and their respective reference strains. The microbiological quality of honeys and the antibiotic susceptibility of the various isolates were also examined. The bioassay applied for determining the antimicrobial effect employs the well-agar diffusion method and the estimation of minimum active dilution which produces a 1 mm diameter inhibition zone. All honey samples, despite their origin (coniferous, citrus, thyme or polyfloral), showed antibacterial activity against the pathogenic and their respective reference strains at variable levels. Coniferous and thyme honeys showed the highest activity with an average minimum dilution of 17.4 and 19.2% (w/v) followed by citrus and polyfloral honeys with 20.8 and 23.8% respectively. Clinical isolates of Staphylococcus aureus subsp. aureus, Escherichia coli, Salmonella enterica subsp. Enterica, Streptococcus pyogenes, Bacillus cereus and Bacillus subtilis were proven to be up to 60% more resistant than their equal reference strains thus emphasizing the variability in the antibacterial effect of honey and the need for further research.     

Palynological characteristics of different unifloral honeys from Greece

Three hundred and twenty-nine Greek honey samples of different botanical and geographical origin were collected and examined by organoleptic evaluation, melissopalynological analysis, measurement of electrical conductivity and colour. The results showed that 208 samples were unifloral with 178 of them representing the main types of unifloral honey produced in Greece; that is fir, pine, chestnut, cotton, orange and thyme honey. All honeys had the sensory characteristics typical of their origin and complied with the electrical conductivity standards set by Council Directive 2001/110/EC.

Fir and pine honeydew honey had a low honeydew element/pollen (HDE/P) ratio and belonged to Maurizio's Classes II or III. The pollen types identified in these honeys ranged from 11 to 45%. Chestnut nectar honey contained >90% chestnut pollen, had a total number of plant elements of >245,000/10?g, and low pollen diversity. Cotton honey contained 1.2 to 16.5% cotton pollen, belonged to Maurizio's Class II, and had 22 pollen types, with Castanea sativa L. present in all samples. Orange honey contained 2.9 to 26.5% Citrus spp. pollen, belonged to Maurizio's Class II, and was characterized by the presence of Brassicaceae, Fabaceae, Olea europea L., Quercus coccifera L. and Rosaceae. In thyme honeys Thymus capitatus Hoffm. &; Link. pollen was secondary or predominant ranging from 18.3 to 69.3%. These honeys belonged to Maurizio's Classes I or II and contained greater than 30 pollen types. Other Lamiaceae, Hypericum spp., Brassicaceae, Fabaceae, Rosaceae, and Cistus spp. pollen types appeared in the greatest number of thyme samples.     

Diversity of honey stores and their impact on pathogenic bacteria of the honeybee,Apis mellifera

Honeybee colonies offer an excellent environment for microbial pathogen development. The highest virulent, colony killing, bacterial agents are Paenibacillus larvae causing American foulbrood (AFB), and European foulbrood (EFB) associated bacteria. Besides the innate immune defense, honeybees evolved behavioral defenses to combat infections. Foraging of antimicrobial plant compounds plays a key role for this “social immunity” behavior. Secondary plant metabolites in floral nectar are known for their antimicrobial effects. Yet, these compounds are highly plant specific, and the effects on bee health will depend on the floral origin of the honey produced. As worker bees not only feed themselves, but also the larvae and other colony members, honey is a prime candidate acting as self‐medication agent in honeybee colonies to prevent or decrease infections. Here, we test eight AFB and EFB bacterial strains and the growth inhibitory activity of three honey types. Using a high‐throughput cell growth assay, we show that all honeys have high growth inhibitory activity and the two monofloral honeys appeared to be strain specific. The specificity of the monofloral honeys and the strong antimicrobial potential of the polyfloral honey suggest that the diversity of honeys in the honey stores of a colony may be highly adaptive for its “social immunity” against the highly diverse suite of pathogens encountered in nature. This ecological diversity may therefore operate similar to the well‐known effects of host genetic variance in the arms race between host and parasite.     

Traditional melissopalynology integrated by multivariate analysis and sampling methods to improve botanical and geographical characterisation of honeys

Abstract

Melissopalynological analysis is the official test to determine the botanical and geographical origin of honey. Geographical characterisation is generally obtained by synthesising the pollen data of all available local honeys in a pollen spectrum interpreted according to palynological expertise. The subjective ability to interpret data is considered a limiting factor for this method and alternative tests are promoted. Nevertheless, melissopalynological analysis is still the most reliable test, and data, if properly exploited, could provide a large amount of information that is generally disregarded. The aim of our work was to apply a new experimental approach for honey sampling, and a statistical multivariate analysis of melissopalynological data. The study was conducted in the Sorrento-Amalfi Peninsula (southern Italy) where 36 hives were positioned in 12 experimental stations distributed in three main land-use types (citrus orchards, chestnut woods and Mediterranean maquis). The new approach proved to be useful for the comprehensive evaluation of honey variability across time and space, allowing objective identification of the pollen types as markers of a given geographical area.     

A variety of volatile compounds as markers in unifloral honey from dalmatian sage (Salvia officinalis L.)

Volatile compounds of unifloral Salvia officinalis L. honey has been investigated for the first time. The botanical origin of ten unifloral Salvia honey samples has been ascertained by pollen analysis (the honey samples displayed 23-60% of Salvia pollen). Fifty-four volatile compounds were identified by GC and GC/MS in ten Salvia honey extracts obtained by ultrasound-assisted extraction (USE) with pentane/Et(2)O 1 : 2. The yield of isolated volatiles varied from 25.7 to 30.5 mg kg(-1). Salvia honey could be distinguished on the basis of the high percentage of benzoic acid (6.4-14.8%), and especially phenylacetic acid (5.7-18.4%). Minor, but floral-origin important volatiles were identified such as shikimate pathway derivatives, 'degraded-carotenoid-like' structures (3,5,5-trimethylcyclohex-2-ene derivatives) and 2,6,6-trimethylcyclohex-2-ene derivatives. Compounds from other metabolic pathways such as aliphatic acids and higher linear hydrocarbons, as well as heterocycles (pyrans, furans, and pyrroles), were also present. Most of the identified compounds do not constitute specific Salvia honey markers, due to their presence in honeys of other botanical origins; however, their ratio in different honeys could be useful to distinguish floral origin. Salvia-honey volatile markers were: benzoic acid, phenylacetic acid, p-anisaldehyde, alpha-isophorone, 4-ketoisophorone, dehydrovomifoliol, 2,6,6-trimethyl-4-oxocyclohex-2-ene-1-carbaldehyde, 2,2,6-trimethylcyclohexane-1,4-dione, and coumaran.     

Geographical,botanical and chemical profile of monofloral Italian honeys as food quality guarantee and territory brand

Bees represent an important element for the preservation of plant biodiversity. During their activities, bees cover a large area around the hive and their products strictly reflect the distinctive traits of these environments. In honey, the floral diversity of the areas inspected by bees is revealed by their pollen spectra. The present work characterized 460 Italian unifloral honeys by melissopalynological and physicochemical analyses. Obtained data contributed to: confirm the botanical identity of the samples; determine the plant biodiversity revealed in each honey; differentiate the samples of the same monofloral typology in relation to their geographical origin; and identify peculiar floristic associations in the Italian regions. Moreover, gas chromatographic analysis was carried out on rare unifloral honey to study their sugar profiles. This work provided the scientific base to create a valid tool able to assign a territorial brand and an authenticity guarantee to the honey, protecting the consumers from adulterations.     

The Effect of New Zealand Kanuka,Manuka and Clover Honeys on Bacterial Growth Dynamics and Cellular Morphology Varies According to the Species

Treatment of chronic wounds is becoming increasingly difficult due to antibiotic resistance. Complex natural products with antimicrobial activity, such as honey, are now under the spotlight as alternative treatments to antibiotics. Several studies have shown honey to have broad-spectrum antibacterial activity at concentrations present in honey dressings, and resistance to honey has not been attainable in the laboratory. However not all honeys are the same and few studies have used honey that is well defined both in geographic and chemical terms. Here we have used a range of concentrations of clover honey and a suite of manuka and kanuka honeys from known geographical locations, and for which the floral source and concentration of methylglyoxal and hydrogen peroxide potential were defined, to determine their effect on growth and cellular morphology of four bacteria: Bacillus subtilis, Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa. While the general trend in effectiveness of growth inhibition was manuka>manuka-kanuka blend>kanuka>clover, the honeys had varying and diverse effects on the growth and cellular morphology of each bacterium, and each organism had a unique response profile to these honeys. P. aeruginosa showed a markedly different pattern of growth inhibition to the other three organisms when treated with sub-inhibitory concentrations of honey, being equally sensitive to all honeys, including clover, and the least sensitive to honey overall. While hydrogen peroxide potential contributed to the antibacterial activity of the manuka and kanuka honeys, it was never essential for complete growth inhibition. Cell morphology analysis also showed a varied and diverse set of responses to the honeys that included cell length changes, cell lysis, and alterations to DNA appearance. These changes are likely to reflect the different regulatory circuits of the organisms that are activated by the stress of honey treatment.     

Major plants foraged by bees for honey production in Ghana: mapping of bee floral sources for the development of the apicultural industry

Abstract

Honey is increasingly being used as a food supplement and to treat various diseases and disorders in Ghana. Information on floral nectar preferences of foraging honeybees is, therefore, imperative to increase production to keep pace with demand. However, in Ghana, these floral nectar preferences are yet to be investigated and identified. This study was conducted to determine bee forage sources in the three main agro-ecological honey-producing zones of Ghana. Forty-eight unblended honey samples sourced from beekeepers were analysed. Pollen spectra of the honey samples revealed 27 different pollen types of which eight were identified to family level, while 15 and four were at the level of genus and species, respectively. The results also showed that, in general, 48% of the analysed samples were unifloral honeys with the most important pollen types coming from Sapotaceae/Meliaceae, Anacardiaceae and Burkea africana. Combretaceae, Sapotaceae/Meliaceae and Anacardiaceae, as well as Lannea-type and Burkea africana were found to be the most frequent (with occurrence greater than 50%) sources of nectar utilised by bees to produce honey. The predominant pollen sources of the honeys in the three main apicultural zones of the country were Sapotaceae/Meliaceae and Anacardiaceae (Forest zone), Sapotaceae/Meliaceae and Lannea-type (Savannah–Forest transition zone) and Sapotaceae/Meliaceae, Anacardiaceae and Gynandropsis gynandra (Savannah zones). Identification of the major bee floral sources can be used as a guide by beekeepers in the location of their apiaries as well as reforesting disturbed sites with these plants to act as sources of bee forage.     

Palynological and geographical characterization of avocado honeys in Spain

The present work refers to the pollen analysis of 12 avocado honey samples from Spain. The samples were directly provided by the beekeepers, all professionals. The quantitative analysis showed that nectar is the main honey source in the samples studied, and that most honeys have a medium presence of botanical elements (BE); one sample belong to Class I of Maurizio, seven to Class II and four belong to Class III. The qualitative analysis of the samples showed the presence of 56 taxa belonging to 36 families. The Spanish avocado honeys are characterised by their medium content in pollen grains (NPG; x¯=117000) and their low honeydew indicator elements content (HDE; x¯=6340). Echium plantagineum gr. and Genista f. (present in 90% of the samples), and Eucalyptus f., Olea europaea L., Mentha aquatica gr. and Reseda luteola gr. (present in 80% of the samples), could be mentioned among the characteristic accompanying species of this honey type. The avocado honeys from the Iberian Peninsula and from the Canary Islands can be differentiated by the presence of Asphalthium bituminosum Medic., Cardiospermum grandiflorum Sw., Phoenix canariensis Hort. ex Chabaud and Tropaeolum majus L. in the latter.     

Pollen analysis of honeys from the Mamora forest region (NW Morocco)

The present work refers to the pollen analysis of 35 Moroccan honey samples from the Mamora forest region. The samples were directly provided by the beekeepers, all professionals. The quantitative analysis showed that nectar is the main honey source in the samples studied, and that most honeys have a medium-low presence of botanical elements (BEN). The qualitative analysis of the samples showed the presence of 54 taxa belonging to 29 families, and 31 of the samples were unifloral: 24 of eucalyptus, 3 of orange, 2 of Loeflingia, 1 of mint and 1 of Ridolfia segetum. The eucalyptus honeys of the studied region are characterized by their high content in pollen grains (NGP; x¯=180000) and their low honeydew indicator elements content (HDE; x¯=4000); Plantago f. (present in 70% of the samples), Quercus f. and Brassicaceae (50%) and Ceratonia siliqua (30%) could be mentioned among the characteristic accompanying species of this honey type.     

Honey shows potent inhibitory activity against the bovine testes hyaluronidase

The purpose of this study was to investigate the anti-hyaluronidase activities of honeys from different botanical origins honeys in order to determine their anti-inflammatory properties. The total phenolic contents, total flavonoids and total tannin levels of six types of honey, chestnut, oak, heather, pine, buckwheat and mixed blossom, were determined. Concentration-related inhibition values were tested turbidimetrically on bovine testis hyaluronidase (BTHase) as IC₅₀ (mg/mL). All honeys exhibited various concentration-dependent degrees of inhibition against BTHase. Inhibition values varied significantly depending on honeys’ levels of phenolic contents, flavonoid and tannin. The honeys with the highest anti-hyaluronidase activity were oak, chestnut and heather. In conclusion, polyphenol-rich honeys have high anti-hyaluronidase activity, and these honeys have high protective and complementary potential against hyaluronidase-induced anti-inflammatory failures.     

Effects of Selected Egyptian Honeys on the Cellular Ultrastructure and the Gene Expression Profile of Escherichia coli

The purpose of this study was to: (i) evaluate the antibacterial activities of three Egyptian honeys collected from different floral sources (namely, citrus, clover, and marjoram) against Escherichia coli; (ii) investigate the effects of these honeys on bacterial ultrastructure; and (iii) assess the anti-virulence potential of these honeys, by examining their impacts on the expression of eight selected genes (involved in biofilm formation, quorum sensing, and stress survival) in the test organism. The minimum inhibitory concentration (MIC) of the honey samples against E. coli ATCC 8739 were assessed by the broth microdilution assay in the presence and absence of catalase enzyme. Impacts of the honeys on the cellular ultrastructure and the expression profiles of the selected genes of E. coli were examined using transmission electron microscopy (TEM) and quantitative real-time polymerase chain reaction (qPCR) analysis, respectively. The susceptibility tests showed promising antibacterial activities of all the tested honeys against E. coli. This was supported by the TEM observations, which revealed “ghost” cells lacking DNA, in addition to cells with increased vacuoles, and/or with irregular shrunken cytoplasm. Among the tested honeys, marjoram exhibited the highest total antibacterial activity and the highest levels of peroxide-dependent activity. The qPCR analysis showed that all honey-treated cells share a similar overall pattern of gene expression, with a trend toward reduced expression of the virulence genes of interest. Our results indicate that some varieties of the Egyptian honey have the potential to be effective inhibitor and virulence modulator of E. coli via multiple molecular targets.     

A melissopalynological study of 54 Louisiana (U.S.A.) honeys

Pollen analyses were made of 54 commercial Louisiana (U.S.A.) honeys collected during 1967–1968. Fifty-eight different pollen types were identified. A direct correlation was assumed between a plant's pollen quantity and its nectar contribution. On this basis, the major Louisiana honey plants that were determined are: Fabaceae, predominantly Trifolium repens (99%), Rubus, Berchemia scandens, and Salix. Plants established as important for their nectar contributions in isolated samples are: Cephalanthus occidentalis, Sapium sebiferum, miscellaneous Compositae (Asteraceae), Glycine max, and Parthenocissus quinquefolia. Native plants such as Rubus, Berchemia scandens, and Salix contribute more to Louisiana's honey sources than do cultivated plants or introduced plants. Most samples originated from six to fifteen different plant types and thus do not appear to be of particularly diverse botanical origin. The pollen types that indicate late spring, summer, or fall honeys include: Glycine max, Cassia fasciculata, Polygonum, Lagerstroemia indica, Lythrum, Ampelopsis, Trachelospermum difforme, and others. Thirty-one samples could be considered unifloral honeys. Most Louisiana honeys were a clear color and from nectar of flowers and not honeydew.     

Study of plants visited by honeybees (Apis mellifera L.) in the Central Rif Region (N. Morocco) using pollen analysis

Quantitative pollen analyses of Moroccan honey samples provided by amateur beekeepers from the Central Rif region demonstrates that nectar is generally the main honey source, and that most honeys have a medium‐low presence of botanical elements. However, two samples reached values as high as 1.08–1.83 qualifying as honeydew honeys. Qualitative results yielded 60 pollen types belonging to 33 families. Ammi visnaga L., Mentha pulegium L., M. rotundifolia (L.) Hudson, Eucalyptus camaldulensis Dehnh. and Rubus ulmifolius Schott. were the main nectar sources, and Cistaceae, Poaceae, Cannabis sativa L. and grasses were exploited as a pollen source. When compared to the parent vegetation honeydew and mint sources are widely exploited, but other sources such as eucalyptus and heather are underutilised.     

Botanical characterisation of Mexican honeys from a subtropical region (Oaxaca) based on pollen analysis

Melissopalynological analysis of 39 honey samples from Oaxaca, Mexico, enabled us to establish the important plant sources exploited by bees during the principal harvest in four districts of the State of Oaxaca, Mexico. A total of 64 taxa belonging to 29 families were recorded. These subtropical honeys were characterised by their botanical origin as follows: (a) monofloral honeys of Bursera simaruba, Clethra mexicana, Cordia alliodora, Lonchocarpus sp., Mangifera indica, Miconia argentea, Orbignya cohune and Quercus sp.; (b) bifloral honeys with an association of Heliocarpus donnell-smithii and Ceiba sp., Lonchocarpus sp. and Mimosa pudica, H. donnell-smithii and Mangifera indica, Miconia argentea and Miconia tenuiflora; (c) oligofloral honeys of Asteraceae; and (d) multifloral honeys with three or four species ≥10%. Monofloral honeys were placed in classes I, II, III, IV and V. Oligofloral were class II, bifloral were classes I and II, and polyfloral honeys were assigned to classes I, II and III. Honey samples of Apis mellifera had a diversity index range of 0.3 to 2.7. It is well known that this bee is polylectic and has a heterogeneous foraging behaviour. In the State of Oaxaca, it prefers resources of secondary vegetation from low deciduous forest, although taxa of economic importance were also utilised, for instance, Mangifera indica and Citrus sinensis. Oaxaca has important and diverse native resources, and beekeeping activity needs to be promoted because of its potential to develop new types of honey.     

Palynological evaluation of selected honeys from Romania

Fifty-four honey samples collected over two consecutive harvest seasons (2009–2010) from different floristic areas of Romania were analysed. A melissopalynological analysis including qualitative and quantitative analyses was carried out in order to identify the principal pollen types in Romanian honeys and therefore, the important plants exploited by Apis mellifera in this country. Seventy-seven pollen types from 35 botanical families were identified. The main pollen forms were: Brassica napus-type, Tilia, Helianthus annuus, Robinia pseudoacacia, Prunus, Castanea sativa, Fragaria-type and Plantago-type. Honey samples were classified as: acacia honey (Robinia pseudoacacia), lime honey (Tilia), rape honey (Brassica napus-type), sunflower honey (Helianthus annuus), cherry honey (Prunus). Three of them were honeydew honeys whereas the others were all polyfloral. The results from this study can be used as a palynological baseline data of Romanian artisanal honeys.     

Pollen analysis of honeys from Caatinga vegetation of the state of Bahia,Brazil

Based on pollen analysis, 17 honey samples collected in the Caatinga area from Nova Soure city were classified by botanical origin to identify the most important floral sources. Most of the honey samples were obtained in August and September. A total of 73 pollen types were identified belonging to 30 families, 64 genera and 30 species. The families best represented by their number of pollen types were Mimosaceae (11), Caesalpiniaceae (9), Rubiaceae and Fabaceae (5 each). Predominant pollen types were: Mimosa arenosa in four samples, M. sensitiva and M. tenuiflora in one sample. Pollen from Mimosa ursina was registered for the first time in the pollen spectrum of the Caatinga vegetation. The correspondence analysis showed a similarity among the honey samples based on pollen composition. The high representation of pollen from native species such as Chamaecrista nictitans, C. ramosa, C. swainsonii and Copaifera martii (Caesalpiniaceae); Aeschynomene martii and Zornia sericea (Fabaceae); Herissantia tiubae (Malvaceae); Mimosa arenosa, M. quadrivalvis, M. sensitiva, M. tenuiflora, M. ursina, Piptadenia moniliformis and Plathymenia reticulata (Mimosaceae), and Ziziphus joazeiro (Rhamnaceae) supports the origin of these honeys from Caatinga vegetation.     

Validation of botanical origins and geographical sources of some Saudi honeys using ultraviolet spectroscopy and chemometric analysis

This study aims at distinguishing honey based on botanical and geographical sources. Different floral honey samples were collected from diverse geographical locations of Saudi Arabia. UV spectroscopy in combination with chemometric analysis including Hierarchical Cluster Analysis (HCA), Principal Component Analysis (PCA), and Soft Independent Modeling of Class Analogy (SIMCA) were used to classify honey samples. HCA and PCA presented the initial clustering pattern to differentiate between botanical as well as geographical sources. The SIMCA model clearly separated the Ziziphus sp. and other monofloral honey samples based on different locations and botanical sources. The results successfully discriminated the honey samples of different botanical and geographical sources validating the segregation observed using few physicochemical parameters that are regularly used for discrimination.     

Comparative study of physio-chemical analysis of fresh and branded honeys from Pakistan

Honey is a nutritious substance produced by bees. Its quality and nutritional value is of great importance for consumers. Keeping this in view physicochemical and minerals determination as quality parameters of fresh floral Pakistani honeys produced by A. mellifera and branded honeys was conducted. The results of fresh honey indicated average means of Color as 48.78 mmPfund, pH 4.9, Total acidity 37.14 meq/kg, Moisture content 18.62%, Electrical conductivity 0.23 mS/cm, Ash content 0.49%, HMF content 30.85 mg/kg, Proline 365.84 mg/kg, Diastase activity 34.39(DN) and Invertase activity was 68.61(IN) comparable to honey standards. Natural honey were rich in k+ (408.46 ppm) and Na+ (405 ppm). Although Ca+ was very low. Whereas, Co, Mn and Ba concentrations exceed the 1 ppm. However, Pb, Cr, and Mo were unnoticeable. Similarly, Color, pH, MC, EC, T. Acidity, HMF, Proline, Ash content, Diastase and Invertase activity of branded honey samples average means found were 42.5 mmPfund, 5.05, 20.5%, 0.18 mS/cm, 15.34 meq/kg, 36.5 mg/kg, 181.6 mg/kg, 1.11%, 7.90(DN) and 36.97(IN) respectively. The findings showed that fresh honey samples were good and of consumable quality as per honey standards than branded honey. Higher HMF content and lower enzymatic activity in branded honey sample than the Codex standards revealed its either long or improper storage.