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The honey bee forage resources in the Neretva Valley enable production of unifloral Satsuma mandarin (Citrus unshiu Marc.) honey. Given the insufficient knowledge about this type of honey, the aim of this study was to establish its botanical origin (pollen spectrum), as well as physico-chemical and sensory properties. In total nine samples of Satsuma mandarin unifloral honey collected from Neretva Valley in the period from 2014 to 2015 were analysed. Floristic observations have recorded 78 plant species and the results of qualitative melissopalynological analysis revealed pollen originating from 50 plant species in analysed honey samples. The number of determined types of pollen in individual honey samples varied from 8 to 19 with an average of 13 pollen types. Proportion of Satsuma mandarin pollen ranged from 3 to 35% with average of 10.88%. The average water content in honey samples range from 17.1% to 17.3%, electrical conductivity from 0.22 mS/cm to 0.27 mS/cm, pH value from 3.82 to 4.02 and diastase activity from 7.78 to 9.36. The main sensory attributes of Satsuma mandarin were determined by sensory analysis: colour ranging from light to dark yellow, orange blossom odour with medium intensity, weak to medium taste persistence characterised by medium sweetness and poorly expressed acidity, and floral and fruity aroma. Due to almost monocultural orchards of Satsuma mandarin in the Neretva Valley, the possibility of producing this rare type of honey in this region should certainly be utilized., Na osnovu pašnih resursa u dolini Neretve značajnu proizvodnju uniflornog meda od unšijske mandarine (Citrus unshiu Marc.) omogućavaju površine pod navedenim agrumom. Međutim, zbog nedovoljnih saznanja o ovoj vrsti meda cilj je ovog istraživanja bio utvrditi njegovo botaničko podrijetlo te fizikalno-kemijska i senzorska svojstva. S navedenog je područja analizirano ukupno 9 uzoraka meda od unšijske mandarine prikupljenih u razdoblju od 2014. do 2015. godine. Florističkim je opažanjima zabilježeno 78 biljnih vrsta, a kvalitativnom je melisopalinološkom analizom u uzorcima meda utvrđena pelud 50 biljnih vrsta. Broj se utvrđene peludi po uzorku meda kretao od 8 do 19, a prosjek je bio 13. Udio peludnih zrnaca unšijske mandarine u istraživanim se uzorcima kretao od 3 do 35% s prosjekom od 10.88 %. Prosječni se udio vode u uzorcima meda kretao od 17.1 do 17.3%, električne provodnosti od 0.22 mS/cm do 0.27 mS/cm, pH vrijednosti od 3.82 do 4.02, dok se aktivnost dijastaze kretala od 7.78 do 9.36. Senzorskom je analizom utvrđeno da boja meda varira od svijetlo do tamno žute, miris meda je po parfemu narančina cvijeta i srednjeg je intenziteta, okus je slabo do srednje postojan sa srednje izraženom slatkoćom i slabo izraženom kiselošću, dok je aroma srednje postojana, cvjetna, odnosno voćna. Na osnovu gotovo monokulturnih nasada unšijske mandarine u dolini Neretve trebalo bi iskoristiti mogućnost proizvodnje ove raritetne vrste meda.

Chemical characterization of bee pollen is of great importance for its quality estimation. Multifloral and unifloral bee pollen samples collected from continental, mountain and Adriatic regions of Croatia were analyzed by means of physico-chemical, chromatographic (GC-MS), and spectroscopic (FTIR-ATR) analytical tools, aiming to conduct a comprehensive characterization of bee pollen. The most distinctive unifloral bee pollen with regard to nutritional value was Aesculus hippocastanum (27.26% of proteins), Quercus spp. (52.58% of total sugars), Taraxacumofficinale (19.04% of total lipids), and Prunusavium (3.81% of ash). No statistically significant differences between multifloral and unifloral bee pollen from different regions were found for most of the physico-chemical measurement data, with an exception of melezitose (p = 0.04). Remarkable differences were found among the bee pollen HS VOCs. The major ones were lower aliphatic compounds, monoterpenes (mainly linalool derivatives, especially in Prunusmahaleb and P.avium bee pollen), and benzene derivatives (mainly benzaldehyde in T.officinale and Salix spp.). Aldehydes C9 to C17 were present in almost all samples. FTIR-ATR analysis revealed unique spectral profiles of analyzed bee pollen exhibiting its overall chemical composition arising from molecular vibrations related to major macromolecules—proteins, lipids, and carbohydrates (sugars).

Due to its multifunctional and complex role in the honey bee colony functioning and health (construction material allowing food storage, brood rearing, thermoregulation, mediation in chemical and m...

Beeswax adulteration in the apiculture sector represents a growing problem worldwide due to the lack of clearly defined purity criteria, the absence of official quality (authenticity) controls, and the inconsistency of the analytical methods used for adulteration detection. Although beeswax authentication is implemented in other regulatory sectors (pharmaceutical and food industry), the classical physico-chemical analytical methods used for determination of beeswax purity exhibit inconsistencies for the detection of adulterants. In this study, an inventory was made on a comprehensive set of analytical methods and the corresponding purity criteria used for the detection of the most common beeswax adulterants (paraffin, stearin and/or stearic acid) from existing legislations and scientific literature. The selected analytical methods (classical physico-chemical, and advanced instrumental, i.e. chromatographic and spectroscopic analytical techniques) were weighted by three independent experts against two criteria: feasibility and analytical performance in detecting targeted adulterants. Classical methods for which measurement data were available (melting point and acid/saponification/ester values for paraffin-adulterated vs. non-adulterated beeswax samples) were retained and further validated by a receiver operating characteristic (ROC) analysis. These methods were also validated by generating the corresponding calibration curves for paraffin detection using paraffin-beeswax mixtures containing different proportions of paraffin (ranging from 5 to 95%, w/w). The results of the ROC analysis revealed that a tentative detection of paraffin in beeswax can be achieved by a combination of at least two physico-chemical methods. However, for a reliable detection of the most common adulterants in beeswax, physico-chemical methods should be complemented with advanced analytical tools. i.e. GC-MS, HTGC-FID (MS) and/or FTIR-ATR spectroscopy, depending on the expected adulterant.

The incidence of antibiotic resistance in pathogenic bacteria has become an alarming clinical and social problem. Therefore, the demand for alternative antimicrobial compounds has increased. In this study, a chemical profile of honey bee (Apis mellifera L.) venom (HBV) has been determined by HPLC and FTIR-ATR spectroscopy, and tested for antibacterial activity, as well as efficiency with regard to conventional antibiotics. The investigated HBV was of high quality with melittin and total protein contents of 70.10 ± 7.01%, and 84.44 ± 3.12 g/100 g, respectively. The purity of HBV was confirmed by FTIR-ATR spectral profiling, which revealed a unique pattern of absorption bands that are characteristic of its major fractions. In addition, HBV showed a broad spectrum of activity against all three tested biomasses of potentially pathogenic Gram-positive and Gram-negative bacteria with MIC values ranging between 12.5 and 200 µg/mL, and MBC between 12.5 and 400 µg/mL. When compared to conventional antibiotics, HBV (400 µg) showed up to 27.8% efficiency of tetracycline (30 µg), 52.2% erythromycin (15 µg), 21.2% ciprofloxacin (5 µg), and 34.6% of ampicillin-sulbactam (20 µg). The overall results demonstrate the therapeutic potential of the analyzed HBV.

There is no systematic report about propolis chemical biodiversity from the Adriatic Sea islands affecting its antioxidant capacity. Therefore, the samples from the islands Krk, Rab, Pag, Bi&scaron, evo and Korčula were collected. Comprehensive methods were used to unlock their chemical biodiversity: headspace solid-phase microextraction (HS-SPME) and hydrodistillation (HD) followed by gas chromatography and mass spectrometry (GC-MS), Fourier transform mid-infrared spectroscopy (FT-MIR), ultra high performance liquid chromatography with diode array detector and quadrupole time-of-flight mass spectrometry (UHPLC-DAD-QqTOF-MS) and DPPH and FRAP assay. The volatiles variability enabled differentiation of the samples in 2 groups of Mediterranean propolis: non-poplar type (dominated by &alpha, pinene) and polar type (characterized by cadinane type sesquiterpenes). Spectral variations (FT-MIR) associated with phenolics and other balsam-related components were significant among the samples. The UHPLC profiles allowed to track compounds related to the different botanical sources such as poplar (pinobanksin esters, esters and glycerides of phenolic acids, including prenyl derivatives), coniferous trees (labdane, abietane diterpenes) and Cistus spp. (clerodane and labdane diterpenes, methylated myricetin derivatives). The antioxidant potential determined by DPPH ranged 2.6&ndash, 81.6 mg GAE/g and in FRAP assay 0.1&ndash, 0.8 mmol Fe2+/g. The highest activity was observed for the samples of Populus spp. origin. The antioxidant potential and phenolic/flavonoid content was positively, significantly correlated.

Analytical parameters were determined for the first time in honey produced in the honeycomb constructed on comb foundations adulterated with 90% of paraffin (PF-H) and compared to honey ripened in genuine beeswax (BWF-H) using physico-chemical and spectroscopic techniques (1H NMR, FTIR-ATR, HS-SPME/GC–MS). Water content was significantly higher (SH) and glucose/water ratio significantly lower in PF-H samples. The contents of acetic and citric acid were marginally significantly higher (MSH) in PF-H samples. These findings suggest that adulterated beeswax affects composition of honey as the set of altered parameters indicate chemical changes leaning towards fermentative processes. Moderately changed headspace chemical profile of PF-H honey was determined depending on the floral source (pentanal, α-pinene and benzaldehyde were SH in BWF-H sunflower honey ; butanal was MSH, and 2-phenylethanol was more abundant in BWF-H black locust honey). Higher percentage of nonanal, octane and β-damascenone were found in PF-H samples that could indicate more intensive oxidation.

The European Food Safety Authority (EFSA) develops methodologies and tools for the detection of emerging risks in food and feed. This includes the identification of drivers of emerging risks, such as food frauds, which requires innovative approaches. In this study, an unsupervised machine learning technique called the Latent Dirichlet Allocation (LDA) topic model, was applied on a media corpus in the view of detecting rapidly specific food fraud incidents in the media, i.e. on the Europe Media Monitor Medical Information System (EMM/MEDISYS). LDA topic model can explore large collection of documents discovering the themes associated with the corpus and organize and summarize text documents identifying topics comprised in them, where a topic is defined as a pattern of words with their probability to belong to it. As a specific food fraud incident, beeswax adulteration was taken as an example. Beeswax can be adulterated for financial gain, and, although it is a product from apiculture, it might enter the food chain when it is introduced as honeycomb in honey pots. With the beeswax example, a total of 2276 news articles were retrieved on EMM/MEDISYS and classified into 10 topics showing different levels of relatedness to beeswax adulteration. A manual screening of all articles allowed to validate the classification made by the topic model. The topics that were found the most relevant contained indeed articles on beeswax adulteration incidents reported from official sources. In addition, those topics contained signals of potential emerging risks in the cosmetic and food wrapping sectors. The remaining topics highlighted the emergence of new beeswax market opportunities which supported the identified signals. It is concluded that the LDA topic model can be used to process rapidly information in the media, support the definition of more specific food fraud filters on EMM/MEDISYS and be of direct use for all stakeholders involved in the monitoring, assessment and management of food frauds.

Analytical parameters were determined for the first time in honey produced in the honeycomb constructed on comb foundations adulterated with 90% of paraffin (PF-H) and compared to honey ripened in genuine beeswax (BWF-H) using physico-chemical and spectroscopic techniques (

Although Satsuma mandarin secretes large amounts of nectar and is considered as excellent honey bee plant species, its anthers are either sterile or produce very small amount of pollen. Th erefore, honey bees collect pollen from other plant resources as well. Th e aim of this study was to determine the botanical origin of pollen loads collected with pollen trap during Satsuma mandarin (Citrus unshiu Marc.) honey bee forage, and to investigate relating foraging parameter (pollen collection time). Th e study was carried out on Carniolan honey bee (Apis mellifera carnica Pollmann, 1879) colony during April in 2014 and May 2015 in Neretva Valley, Croatia. Pollen of 12 plant species was identifi ed in analysed samples. Results have revealed that a total pollen mass was signifi cantly higher (p

Experimental varroacidal treatments of honey bee colonies were conducted on five apiaries (EA1&ndash, EA5) situated at five different geographical and climatic locations across Croatia. The aim of this study was to assess the comparative efficacy of CheckMite+ (Bayer, Germany), Apiguard (Vita Europe Ltd., England), Bayvarol, C, (Bayer, Germany), Thymovar, (Andrma BioVet GmbH, Germany), and ApiLife Var, (Chemicals Laif SPA, Vigonza, Italy) for controlling the honey bee obligatory parasitic mite Varroa destructor in different conditions in the field during summer treatment. The relative varroa mite mortality after treatments with applied veterinary medicinal products were EA1 (59.24%), EA2 (47.31%), EA3 (36.75%), EA4 (48.33%), and EA5 (16.78%). Comparing the relative efficacy of applied varroacides, the best effect was achieved with CheckMite+, and the lowest for honey bee colonies treated with Apiguard (statistically significant difference was confirmed, p &lt, 0.05). Considering the lower efficacy of thymol-based veterinary medicinal products observed on all EA in these study conditions, it may be concluded that their use is limited under different treatment regimes. Despite unfavourable weather and environmental conditions, with exceptions of EA5/EA5&prime, and EA1, the relative varroacidal efficacy of authorized veterinary medicinal product treatments in moderately infested honey bee colonies ensured normal overwintering and colony development during next spring.

Samples of Satsuma mandarin (Citrus unshiu Marc.) nectar, honey sac content and honey were analyzed by FTIR-ATR spectroscopy and reference methods. The spectral analysis allowed detection of the major chemical constituents in C. unshiu nectar- to-honey transformation pathway thus providing information on the intensity and location of the compositional changes occurring during this process. The preliminary results showed that in average more than one-third of sugar-related nectar-to-honey conversion takes place directly in the honey sac ; the average sugar content (w/w) was 17.93% (nectar), 47.03% (honey sac) and 79.63% (honey). FTIR-ATR results showed great spectral similarity of analyzed honey samples and small degree variations in both sugar and water content in nectar samples. The spectral data revealed distinctive differences in the chemical composition of individual honey sac contents with the most intensive and complex absorption envelope in the spectral region between 1175 and 950 cm-1 (glucose, fructose and sucrose absorption bands).

Control of the nosema disease poses a major challenge, and therefore, treatment of this serious para - sitic disease using natural preparations could be of great benefit. The aim of this study was to test the performance of zeolite clinoptilolite as a curative measure against honeybee colonies ( Apis mellifera L.) naturally infected by Nosema ceranae. The histopathological structure, and the content and distribution of mucosubstances and his - tochemical activity of aminopeptidase and non-specific esterase in the midgut mucosa of honeybees originating from colonies fed sugar syrup supplemented with zeolite minerals was studied. A decline in the number of spores in honeybees fed with zeolite clinoliptolite was observed on the first sampling day (Day 10; 53.25 15.15 million spores/bee), though a statistically lower number of spores in comparison to the control was confirmed on Day 20 (41.08 ± 9.4 million spores/bee), Day 30 (28.42 ± 7.79 million spores/bee) and Day 40 (24 ± 6.25 million spores/bee). The possibility of using natural zeolites as a dietary supplement for honeybee colonies as a preventative measure and for the reduction of the deleterious effects of nosemosis is discussed.

Satureja hortensis L. unifloral honey was characterized by pollen analysis, electrical conductivity, pH and extensively by chromatographic and spectroscopic techniques. UV / VIS spectro-scopy measurements revealed total phenol content of 682.1 mg GAE / kg by Folin-Ciocalteu assay, antiox-idant capacity by DPPH assay of 1.7 mmol TEAC / kg and by FRAP assay of 4.3 mmol Fe2+ / kg as well as CIE L*a*b*Cab*h°ab chromaticity coordinates. GC-MS after headspace solid-phase microextraction (HS-SPME) revealed hotrienol (22.8 %) along with other linalool derivatives, benzaldehyde (6.1 %), phenylacetaldehyde (4.9 %) and few norisoprenoids (safranal (7.6 %) as the major). Ultrasonic solvent ex-traction (USE) followed by GC-MS allowed identification of methyl syringate (54.7 %) as predominant compound along with other benzene derivatives. HPLC-DAD analysis determined tyrosine (382.0 mg kg−1), phenylalanine (140.4 mg kg−1) and methyl syringate (39.32 mg kg−1). Methyl syringate and hotrienol can be considered non-specific chemical markers of S. hortensis honey. FTIR-ATR spectral characteristics of S. hortensis honey in fingerprinting region were not significantly different from other honey types, but the integrated intensity of the region was smaller than in other unifloral honeys.

Six hundred thirty-two honey samples from Croatia were analyzed by reference methods (physicochemical and melissopalynological analyses), from which 506 were assigned to one of nine unifloral honey types (black locust, sweet chestnut, lime, sage, heath, mandarin, false indigo, immortelle and winter savory). Vibrational spectra of honey samples recorded using Fourier transform infrared (FTIR) spectroscopy and attenuated total reflection (ATR) technique were subjected to comprehensive chemometric assessment involving optimization of data preprocessing and multivariate statistical analyses, namely principal component analysis, partial least squares regression, partial least squares discriminant analysis and receiver operating characteristic (ROC) analysis. A specially designed hybrid multidimensional model (partial least squares-linear discriminnat analysis) was used to assess a relationship between honey IR spectra and relative frequencies of pollen grains (pollen spectrum) present in selected honey samples. The study demonstrated that a targeted spectral profiling, i.e., preprocessing of a fingerprint spectral region (1, 800–700 cm−1) by particular Savitzky–Golay filtering parameters and ROC modelling approach, enables accurate classification (100 % correct classification) of unifloral honeys by their botanical origin with low misclassification risk (MR = 0.099) and negligible small number of misclassified samples (1–2 per class—honey type). The results have also revealed good predictive strength of FTIR-ATR spectroscopy for melissopalynologycal data.

Although beeswax adulteration represents one of the main beeswax quality issues, there are still no internationally standardised analytical methods for routine quality control. The objective of this study was to establish an analytical procedure suitable for routine detection of beeswax adulteration using FTIR-ATR spectroscopy. For the purpose of this study, reference IR spectra of virgin beeswax, paraffin, and their mixtures containing different proportions of paraffin (5 - 95%), were obtained. Mixtures were used for the establishment of calibration curves. To determine the prediction strength of IR spectral data for the share of paraffin in mixtures, the Partial Least Squares Regression method was used. The same procedure was conducted on beeswax-beef tallow mixtures. The model was validated using comb foundation samples of an unknown chemical background which had been collected from the international market (n = 56). Selected physico-chemical parameters were determined for comparison purposes. Results revealed a strong predictive power (R2 = 0.999) of IR spectra for the paraffin and beef tallow share in beeswax. The results also revealed that the majority of the analysed samples (89%) were adulterated with paraffin; only 6 out of 56 (11%) samples were identified as virgin beeswax, 28% of the samples exhibited a higher level of paraffin adulteration (>46% of paraffin), while the majority of the analysed samples (50%) were found to be adulterated with 5 - 20% of paraffin. These results indicate an urgent need for routine beeswax authenticity control. In this study, we demonstrated that the analytical approach defining the standard curves for particular adulteration levels in beeswax, based on chemometric modelling of specific IR spectral region indicative for adulteration, enables reliable determination of the adulterant proportions in beeswax.

There is no report on the chemical composition of alder buckthorn (Rhamnus frangula L.) honey and therefore volatile organic compounds (VOCs) were investigated for the first time as well as their composition after short-term heating. The electrical conductivity of the samples was 0.90 - 0.98 mScm-1 (specificity of alder buckthorn honey), and pH values were 4.00 - 4.33. The melissopalynological analysis revealed R. frangula L. as dominant plant species (52-93%) in all samples. Accompanying minor plant species were also determined (Castanea sativa L., Centaurea jacea L., Lotus corniculatus L., Prunus spp., Salix spp., Apiaceae, Asteraceae, Acer spp. and Cornus sanguinea L.). The VOCs from R. frangula L. honey were isolated by headspace solid-phase microextraction (HS-SPME) and ultrasonic solvent extraction (USE) and analysed by GC-FID and GC-MS. To investigate the influence of short-term heating, selected sample was heated 3h at 100 oC with and without H2O addition. The most representative chemical profile of R. frangula L. honey is obtained by HS-SPME containing benzaldehyde, hotrienol and 4-ketoisophorone. USE contributed to the honey characterization, mainly by vomifoliol identification. The majority of compounds found in USE were higher aliphatic alcohols. The influence of short-term heating to R. frangula L. honey is strongly dependent on H2O addition. Namely, more artefacts (furan derivatives, particularly 5-hydroxymethylfurfural) were formed during heating of the honey dissolved in H2O in comparison with direct honey heating.

Headspace solid-phase microextraction (HS-SPME) and ultrasonic solvent extraction (USE), followed by GC-MS/FID, were applied for monitoring the nectar (NE)/honey-sac (HoS)/honey (HO) pathways of the headspace, volatiles, and semi-volatiles. The major NE (4 varieties of Citrus unshiu) headspace compounds were linalool, -terpineol, 1H-indole, methyl anthranilate, and phenylacetonitrile. Corresponding extracts contained, among others, 1H-indole, methyl anthranilate, 1, 3-dihydro-2H-indol-2-one and caffeine. The major HoS headspace compounds were linalool, -terpineol, 1, 8-cineole, 1H-indole, methyl anthranilate, and cis-jasmone. Characteristic compounds from HoS extract were caffeine, 1H-indole, 1, 3-dihydro-2H-indol-2-one, methyl anthranilate, and phenylacetonitrile. However, HO headspace composition was significantly different in comparison to NE and HoS with respect to phenylacetaldehyde and linalool derivatives abundance that appeared as the consequence of the hive conditions and the bee enzyme activity. C. unshiu honey traceability is determined by chemical markers: phenylacetaldehyde, phenylacetonitrile, linalool and its derivatives, as well as 1H-indole, 1, 3- dihydro-2H-indol-2-one, and caffeine.

Pčelarima služi na ponos spominjanje pčele u Vinodolskom zakoniku koji je proglašen 6. siječnja 1288. godine, točno prije 735. godina. To je najstariji u cijelosti sačuvan spomenik običajnog prava na hrvatskom jeziku pisan glagoljicom na čakavskom dijalektu. Vrlo je teško novim internet generacijama, objasniti da mi dominantna „ljudska populacija“ ovisimo o tim malim korisnim kukcima oprašivačima. Internet generacija koja dominira na Planetu Zemlji i ponaša se kao najveći egocentrik i želi sve sebi prilagoditi ne shvaća da je čovjek postao čovjeku najveći neprijatelj. Većina njih zna da ih pčela može „piknuti“ i izazvati bol, odnosno da pčele stvaraju med. Iz toga razloga oni se s pravom „boje“ pčela, ali se raduju medu. Od dvjestotinjak važnih poljoprivrednih kultura koje se uzgajaju na području Europske unije njih 80 % izravno ovisi o oprašivanju korisnim kukcima (medonosne pčele, bumbari, divlje i solitarne pčele). Tijekom prošlog stoljeća u svijetu je za približno 50 % smanjen broj uzgajanih pčelinjih zajednica, ali u isto vrijeme je za više od 300 % povećana proizvodnja poljoprivrednih kultura koje izravno ovise o oprašivanju korisnim kukcima. Širom planeta Zemlja malo što kao pčele, bez obzira na vjeru, spol, boju kože, obrazovanje povezuje ljude. Priče o pčelama su bile i biti će nikada ispričane priče, svi smo mi bar jednom u životu imali bliski susret s pčelama. Veza između pčela i čovjeka danas je neraskidiva, a u sadašnje vrijeme izložena je čitavom nizu opasnosti koje smo mi ljudi stavili pred pčele nepravilnim korištenjem raznih „cida“. Broj pčela u okolišu je primarni indikator opstojnosti Planeta. Ako danas nestane pčela za nekoliko godina na Planeti će nestati i ljudske populacije. Iz toga razloga u Europskom zelenom planu koji će se provoditi do 2050. godine pčele imaju važno mjesto i ulogu (Revizija Inicijative EU-a za oprašivače). Kroz ovaj skup želimo naglasiti nezamjenjivo mjesto i ulogu pčela u okolišu.