Beeswax Samples Research Articles

Evaluation of environmental contamination in beeswax products

Beeswaxes are used as a coating agent or as a wrapping material for food products making them potentially ingested by consumers. There is no regulation yet in Europe giving maximum levels of contaminants in this type of product. Nevertheless, being a natural product, they are exposed to environmental pollution, thus it appears necessary to establish their contamination rate in order to evaluate potential human exposure. In this study, a method of extraction of different environmental contaminants including pesticides, phthalates, PAHs and phenols was developed. Based on a hot Soxhlet extraction, followed by cleaning steps, the method was validated for the quantitation of the cited contaminants by LC-MS/MS and GC-(MS)/MS.Three different types of waxes were analyzed including typical white waxes (Cera Alba) and yellow waxes (Cera Flava). It was shown that all waxes had the presence of at least one contaminant and that phthalates, in particular DEHP, was present in all beeswax samples. Insecticides were found in majority among all the classes of pesticides screened. The yellow waxes were found to be contaminated with the highest rates of PAHs (60%), pesticides (75%) and phenols (40%). The detection frequency of PAHs, in contrast to phthalates, was the lowest for all the types of waxes combined.

INVESTIGATION OF HEAVY METAL LEVELS OF BEESWAX IN BEE APIARIES IN ÇANKIRI

A total of 25 samples of beeswax (9 from the southern and 16 from the northern of Çankırı) were collected within the scope of this study. Heavy metal contents of these samples were analyzed by using inductively coupled plasma mass spectroscopy (ICP-MS). The mean iron (Fe), zinc (Zn), copper (Cu), manganese (Mn), lead (Pb), aluminum (Al), chromium (Cr), vanadium (V), cadmium (Cd), selenium (Se) levels in the samples were found to be 36.28 ppm, 1.69 ppm, 1.65 ppm, 1.14 ppm, 0.78 ppm, 0.68 ppm, 0.30 ppm, 0.13 ppm, 0.06 ppm, 0.01 ppm, respectively. Comparing the northern and southern regions, only Cd (1.10 ppm) was found to be higher in the southern region, whereas the other heavy metals were found to be higher in the northern region. No statistical difference was found between mean Fe, Pb, Zn, Al, Cr, V, and Se levels in the wax samples obtained from the northern and southern regions, whereas a statistically significant difference was found in Mn, Cu, and Cd concentrations. As a result, the heavy metal concentration in beeswax varied between the regions. Except for Cd element, all values were higher in the northern region of Çankırı and this region that is close to the highway with heavy traffic should be evaluated in terms of beekeeping.

Effects of clove bud oil on the physicochemical properties and oxidative stability of canola oil organogels structured by different beeswax contents.

The aim of this study was to investigate the utilization of clove bud oil as fat-soluble antioxidants for retarding lipid oxidation in organogels by structuring canola oil with beeswax at 5, 7.5, and 10% concentration under accelerated oxidation condition. Oil binding capacity and viscoelastic properties were increased with beeswax content, but were not nearly affected by the addition of clove bud oil. Organogel loaded with clove bud oil were found to be more effective in retarding lipid oxidation in high beeswax content systems, particularly evident in 10% beeswax samples. The addition of clove bud oil resulted in low levels of hyeoperxide and MDA, and protected against texture and color deterioration during the storage period. Additionally, the Pearson correlation between lipid oxidation indices and parameters of texture and color has been found to exhibit a limited association, with the exception of the a* and b* values, which show a strong correlation.

Seasonal screening of pesticide residues in beehive products collected from different districts in Egypt

Pesticides are of immense importance in agriculture, but they might contaminate bees’ products. In this study, samples of honey, pollen, and beeswax were collected, seasonally, from apiaries in Toshka (Aswan), El-Noubariya (El-Beheira), and Ismailia (Ismailia) cities in Egypt. The pesticide residues were analyzed using the GC-MS after being extracted and cleaned using the QuEChERS method. Results showed that samples from El-Noubariya had great content of residues followed by Ismailia, and finally Toshka. Samples collected during fall and winter had the highest pesticide residue contents. Specifically, the phenylconazole fungicide group was repeatedly detected in all the examined samples along with organophosphate insecticides. Beeswax samples had the greatest amounts of pesticide residues followed by pollen and then honey samples. Chlorpyrifos (0.07–39.16 ng/g) and profenofos (1.94–17.00 ng/g) were detected in honey samples and their products. Pyriproxyfen (57.12 ng/g) and chlorpyrifos-methyl (39.16 ng/g) were detected in great amounts in beeswax samples from Ismailia and El-Noubariya, respectively. Yet, according to health hazard and quotient studies, the amounts of pesticides detected in honey do not pose any health threats to humans.

Determination of bisphenols in beeswax based on sugaring out–assisted liquid–liquid extraction: Method development and application in survey, recycling and degradation studies

The methodology of sugaring out–assisted liquid–liquid extraction (SULLE) coupled with high-performance liquid chromatography–fluorescence detection was devised for quantifying bisphenol A (BPA) and bisphenol B (BPB) in beeswax. The effectiveness of SULLE was methodically explored and proved superior to the salting out–assisted liquid–liquid extraction approach for beeswax sample preparation. The analytical performance underwent comprehensive validation, revealing detection limits of 10 μg/kg for BPA and 20 μg/kg for BPB. The method developed was employed to analyse commercial beeswax (n = 15), beeswax foundation (n = 15) and wild-build comb wax (n = 26) samples. The analysis revealed BPA presence in four commercial beeswax samples and three beeswax foundation samples, with the highest detected residue content being 88 ± 7 μg/kg. For BPB, two beeswax foundation samples were positive, with concentrations below the limits of quantification and 85 ± 4 μg/kg, respectively. No bisphenols were detected in wild-build comb wax. Furthermore, the bisphenol removal efficacy of two recycling methods—boiling in water and methanol extraction—was assessed. The findings indicated that after four recycling cycles using water boiling, 9.6% of BPA and 29.2% of BPB remained in the beeswax. Whereas methanol extraction resulted in approximately 7% residual after one recycling process. A long-term study over 210 days revealed the slow degradation of bisphenols in comb beeswax. This degradation fitted well with a first-order model, indicating half-lives (DT50) of 139 days for BPA and 151 days for BPB, respectively. This research provides the first report on bisphenol contamination in beeswax. The low removal rate during the recycling process and the gradual degradation in beeswax underscore the significance of bisphenol contamination and migration in bee hives along with their potential risk to pollinators warranting concern. Furthermore, the developed SULLE method shows promise in preparing beeswax samples to analyse other analytes.

Evaluation of the potential migration of acaricides from stamped beeswax to honey simulating beehive conditions: A pilot study

There is an issue concerning the migration of pesticides from stamped beeswax to other bee products, mainly, honey. This implies that honey and other beehive-based products could be contaminated, which would represent a potential risk to consumers and food safety. To determine the degree to which acaricides can be transferred from the stamped beeswax to the honey, a pilot experiment was designed consisting of placing beeswax that contained acaricides (endogenous content, τ-fluvalinate, chlorfenvinphos, and coumaphos; spiked samples, coumaphos) in contact with multifloral honey in an incubator. Temperature and shaking conditions were settled simulating beehive conditions. The isolation of the analytes from beeswax involved the use of a QuEChERS (quick, easy, cheap, effective, rugged & safe) method, and honey was analyzed through a solvent extraction procedure. Acaricides were determined in the resulting extracts by gas chromatography with mass spectrometry detection. Coumaphos was the only acaricide transferred into honey in beeswaxes with endogenous acaricide content; meanwhile, results of the experiments with spiked beeswax samples showed that the initial concentration present in the contaminated beeswax significantly influences the transfer to honey, as it was observed that in the spiked beeswax sheets with high levels of coumaphos, the migration from beeswax to honey arose.

Effect of Storage Time and Floral Origin on the Physicochemical Properties of Beeswax and the Possibility of Using It as a Phase Changing Material in the Thermal Storage Energy Technology

Beeswax is a natural product that is primarily produced by honey bees of the genus Apis. It has many uses in various kinds of industries, including pharmacy and medicine. This study investigated the effect of storage and floral origin on some physicochemical properties of four beeswax samples. The floral origin of the beeswax samples was determined microscopically and the investigated physical properties were the melting point, color, surface characteristics and thermal behavior. The studied chemical constituents were the acid value, ester value, saponification value and the ester/acid ratio. The FT-IR, SEM, EDX, XRD and TGF techniques were applied to meet the objectives of this study. The physical properties of the beeswax were affected by the storage period and floral origin. The melting point of the beeswax samples significantly increased with the increase in the storage time, from 61.5 ± 2.12 °C for the 3 month sample to 74.5 ± 3.54 °C for the 2 year stored sample (p-value = 0.027). The acid values of the 3 month, 6 month, 1 year and 2 years stored samples were 19.57 ± 0.95, 22.95 ± 1.91, 27 ± 1.91 and 34.42 ± 0.95 mgKOH/g, respectively. The increase in the acid value was significant (p-value = 0.002). The ester values of the studied beeswax samples significantly increased with the increase in storage time as follows: 46.57 ± 2.86 mgKOH/g for the 3 month stored sample, 66.14 ± 3.82 mgKOH/g for the 6 month stored sample, 89.77 ± 0.95 mgKOH/g for the one year stored sample and 97.19 ± 1.91 mgKOH/g for the 2 year stored sample (p-value ≤ 0.001). Similarly, the saponification value and the carbon percentages increased with the increase in storage time. Unlike the results of the chemical components, the oxygen percentage decreased with the increase in storage time as follows: 11.24% (3 month), 10.31% (6 month), 7.97% (one year) and 6.74% (two year). The storage and floral origin of beeswax significantly affected its physicochemical properties in a way that qualify it to act as a phase changing material in the thermal storage energy technology.

Quantitation of 1,4-Dichlorobenzene and Thymol in Beeswax Using Dynamic Headspace Vacuum Transfer in Trap Extraction Prior to Gas Chromatography-Mass Spectrometry

A method based on dynamic headspace vacuum transfer in trap extraction, followed by gas chromatography coupled with a mass spectrometer (DHS-VTT-GC-MS), was validated for the fast quantitation of 1,4-dichlorobenzene (p-dichlorobenzene; PDCB) and thymol residues in beeswax. The quantitation limits (LOQ) were 0.05 mg/kg (PDCB) and 0.25 mg/kg (thymol). Recoveries above 80% were obtained for PDCB concentrations between 0.05 and 10 mg/kg and for thymol concentrations between 0.25 and 200 mg/kg. Analysis of beeswax samples showed a good correlation between the results obtained by DHS-VTT-GC-MS analysis and those of a previous method based on SPE extraction followed by gas chromatography and triple- quadrupole mass spectrometry (GC-MS/MS) (R2 = 0.9770 for PDCB and 0.9666 for thymol). However, the sample preparation and chromatography were much shorter using the DHS-VTT-GC-MS method. Forty comb foundations samples produced in Switzerland in 2019 and 2021 were analysed using DHS-VTT-GC-MS. Fourteen samples contained PDCB above the LOQ of 0.05 mg/kg, ranging up to a maximum of 1.53 mg/kg. The mean concentration of the positive samples was 0.22 mg/kg. All samples contained thymol residues ranging between 3.9 and 84.4 mg/kg with mean and median concentrations of 22.7 mg/kg and 17.4 mg/kg. Residue levels of PCDB and thymol in Swiss beeswax were substantially below those measured 20 (PDCB) and 10 (thymol) years ago, respectively.

Suitable Areas for Apiculture Expansion Determined by Antioxidant Power, Chemical Profiles, and Pesticide Residues in Caldcluvia paniculata Honey and Beeswax Samples

Forty-two samples of Tiaca Honey (Caldcluvia paniculata) obtained from beehives belonging to 14 apiaries (three honey samples per apiary) were collected at the end of January near Osorno (40°34' S, 73°8' W), Puyehue (40°40' S, 72°37' W) and Frutillar 41°7' S, 72°59' W) covering an area of 1240 km2. They presented the highest phenol contents (0.36 mg gallic acid equivalent/kg) and antioxidant power (1.27 mM equivalent of Fe+2/g of sample), and were among the highest for antiradical activity. Phenol contents and antioxidant power (r = 0.72, p-value < 0.01) and total phenol contents and antiradical activity (r = 0.69; p-value < 0.01) displayed linear correlations. Only two beeswax samples showed residues of the pesticide fenhexamid. The respective sites (Purranque [40°55' S, 73°10' W] and Coligual [40°49' S, 72°54' W]) were the only areas located near active farms. Additionally, the m/z value 163.1091 was found as an element to identify honeys. Data were used to construct a mapped suitability index ranking for pesticide-free areas with high biological quality. The provided chemical profiles will aid local beekeepers in obtaining international certifications, particularly for the EU market. In turn, the constructed maps indicate suitable areas for apiculture expansion, while differentiated pesticide detection in honey and beeswax requires further comparative research.

Adulteration of beeswax: A first nationwide survey from Belgium.

Beeswax is intended for use in the beekeeping sector but also in the agro-food, pharmaceutical or cosmetics sectors. The adulteration of beeswax is an emerging issue that was reported lately at several occasions in the scientific literature. This issue tends to become more frequent and global, but its exact extent is not accurately defined. The present study aims to assess the current situation in Belgium through a nationwide survey. Randomized beeswax samples originating from Belgian beekeepers (N = 98) and commercial suppliers (N = 9) were analysed with a Fourier transform infrared spectroscopy (FTIR) coupled with Attenuated Total Reflectance (ATR) accessory (FTIR-ATR spectroscopy) for adulteration. The survey revealed a frequency of 9.2% and 33.3% of adulteration in beekeepers beeswax samples (9 samples out of 98: 2 with paraffin and 7 with stearin/stearic acid) and commercial beeswax samples (3 samples out of 9: all adulterated with stearin/stearic acid), respectively. The analysed samples were adulterated with various percentages of paraffin (12 to 78.8%) and stearin/stearic acid (1.2 to 20.8%). This survey indicates that in the beekeepers samples, beeswax adulteration was more frequent in comb foundation and crude beeswax than in comb wax. With the example of this nationwide survey conducted in Belgium, this study shows the emergence of the issue and the urgent need for action to safeguard the health of both honey bees health and humans, in particular with the setting of a proper regulation legal framework and a specific routine analytical testing of commercial beeswax to ensure beeswax quality.

Presence, persistence and distribution of thymol in honeybees and beehive compartments by high resolution mass spectrometry

Thymol has been considered as an organic varroacide with less toxicity than synthetic chemicals such as amitraz, coumaphos or tau-fluvalinate. The present study evaluates the presence, persistence and distribution of thymol during a five-month sampling period; before, during and after the application of thymol treatment in field conditions. Moreover, its impact on the development of bee brood was also studied at different development stages which consist of larvae, prepupae, pupae and adult honeybee. Samples of beebread (processed pollen), honey, beeswax, adult bees, pupae, prepupae and larvae were analysed by gas chromatography coupled to time of flight mass spectrometry for an accurate identification. Adult bees were the samples with the highest concentration level of thymol during treatment application (Thymovar® strips, 30 gr of thymol per hive) by the beekeeper, however they showed 69% of thymol dissipation three months after treatment. Regarding the bee brood, concentrations of thymol decreased significantly (89-97% of disappearance) in a period of three months, though about 42 µg/kg of thymol persisted in larvae. Thymol was found to accumulate in honey and beeswax at a level of 882 μg/kg and 1305 μg/kg respectively, in the studied period. As the main issue of thymol residues in honey is related to the modification of organoleptic properties, more than toxicology, the levels found in this study indicate that the taste of honey may be affected during and up to three months after the treatment application. In addition, we did not find adverse effects on the evolution of the colonies due to the applied treatment.

Removal of pesticide residues from beeswax using a methanol extraction-based procedure: A pilot-scale study

In the present study, a batch mode procedure has been developed to remove pesticides from beeswax coming from bee farms. The beeswax is treated by an Extraction Process with methanol at 65 °C followed by an Adequation Process with water at 70 °C. This procedure can reduce the beeswax contamination load by values of around 80%–90% for the acaricides usually present in beeswax such as Coumaphos, Chlorfenvinphos, tau-Fluvalinate and Acrinathrin, at concentration ranges of 1800–2600 μg/kg, 140–300 μg/kg, 100–300 μg/kg and 160–470 μg/kg, respectively. This process is also able to eliminate other pesticides, such as Amitraz, which are typically present in beeswax samples at high concentrations. Similarly, it is efficient in removing further compounds such as Propiconazole, Bromopropylate, Chlorpyrifos, Cypermethrin, Diazinon and Fenamiphos, which are present at lesser quantities depending on their origin and surrounding pesticide treatment history. The procedure’s operating parameters were atmospheric pressure, a 20 °C–70 °C temperature range and 200 rpm. Up to 80% of the methanol used in the extraction was recycled and the waste generated was easy to manage. The quality of the decontaminated beeswax obtained has similar properties to those of virgin beeswax and can be used to manufacture ecological hives or galenic formulations. Beeswax quantities of 1, 20 and 75 kg were treated to determine the optimal operating conditions that would make it possible to extend the process to the industrial scale. All the equipment utilized (e.g. agitated tanks, temperature control, centrifuges and distillation equipment) are basic tools commonly used in the chemical industry.

Physico-chemical Analysis of beeswax procured from Mumbai market, Western Ghats (Natural Habitat)

Beeswax is the substance that is obtained from the structure of a honey comb. The bees secrete wax to build the honey combs to store honey. It has rich hydrophobic property, the beeswax is in fact present within cosmetics and body products. Also, beeswax is used in food industry. Beeswax is a complex mixture of hydrocarbons, free fatty acid, esters of fatty acid. Beeswax plays an important role in Ayurvedic medicine, the traditional medicine name with Madhushistha (Beewax). The Madhushistha is used for the care of wound from abrasion or even from burns with topical application. Some references in Ayurvedic granth highlight the use of Madhushistha in combination with other herbal and mineral mixture, like Sikta Taila, Mruddaharshringa, Jatyadi Taila, etc. Beeswax is mainly of two types, natural beeswax and processed beeswax. Natural beeswax is clarified further heating in water with petroleum waxes. Because of high demand and shortage in market, adulteration of beeswax with cheaper materials are common like animal fat, plant oil and petroleum spirits (Paraffin wax). Hence authenticity of beeswax is major concern and that can be determined by using physico-chemical parameters such as- organoleptic test, solubility, Melting point, Spacific gravity, Refractive index, Acid value, Ester value, Saponification value, volatile matter. With this justification the present study is expected to determine the quality status of four different beeswax samples procured from Mumbai market, western ghats (Natural habitat). The physico-chemical analysis revealed that volatile matter is detected in market sample (2) whereas market sample (1) shows higher melting point as compare to beeswax collected from the Natural habitat. Market sample (1,2) of beeswax shows slight adulteration hence natural habitat samples (1,2) of beeswax should be used in medicine preparation.

Validation of analytical methods for the detection of beeswax adulteration with a focus on paraffin

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 use of infrared spectroscopy and thermal analysis for the quick detection of adulterated beeswax

Beeswax samples of different origins were evaluated for their authenticity by infrared spectroscopy and thermal analysis. The Fourier Transform Infrared Spectroscopy (FT-IR) method was used for the genuineness assessment and the melting process was examined using the differential scanning calorimetry (DSC) method. Eight suspicious beeswax samples delivered by local beekeepers were examined. As reference standards, pure authentic beeswax standards, pure paraffin, and wax-paraffin blends were used. The comparison of the infrared spectra of tested wax samples and references allowed the identification of adulterated wax samples based on the differences in the spectra fingerprint region (700 to 1800 cm−1), especially 1170 and 1735 cm−1 bands. Moreover, the ratio of the 1735 and 2850 cm−1 band area (lower than 0.5) was used as an indicator of wax adulteration. Among the 8 samples studied, 3 were found to have been falsified with paraffin. The thermal analysis confirmed the adulteration of the same wax samples. The melting temperatures determined in these cases were lower (52.3 to 57.7) as compared to other samples (63.1 to 65.9), identified as real beeswax by comparison to standards (63.8 to 64.7). A similar tendency during the re-heating of cooled wax samples was observed, which suggested the addition of paraffin or another substance with a lower melting point to adulterated waxes. The methods used in this research (FT-IR and DSC) allowed for a relatively quick, easy, and inexpensive verification of the authenticity of the beeswax. Such a method can be used as a screening step before a comprehensive GC analysis of beeswax.

Acetonitrile-Ethyl acetate based method for the residue analysis of 373 pesticides in beeswax using LC-MS/MS and GC–MS/MS

In the current study, Pesticide residue extraction in beeswax was carried out using a mixture of acetonitrile-ethyl acetate (1:3, v/v). This mixture of solvents not only enables the melting of beeswax sample at a lower temperature than when using acetonitrile only but also introduces one phase solution. The sample extract was directly injected into both GC–MS/MS, of the commonly used split-less inlets, and into LC-MS/MS. Sample preparation and clean-up were also optimized. The developed method was validated according to SANTE/11813/2017 European Union guidelines. Three spiking levels of low concentrations 20, 50, 100 µg/kg were studied for the analysis of a total of 373 pesticides. Most of the studied pesticides have acceptable recovery between 80 and 110% with good reproducibility <10. There are 265 and 139 pesticides having a lower limit of quantifications equal 20 µg/kg using LC-MS/MS and GC–MS/MS, respectively. Finally, the developed method was successfully applied for the analysis of real beeswax samples.

Detection of flumethrin acaricide residues from honey and beeswax using high performance liquid chromatography (HPLC) technique

Residues of acaricide, flumethrin were detected from honey and beeswax using high performance liquid chromatography (HPLC) technique. Forty samples of honey were collected before flumethrin treatment and 30, 60, and 90 days after application from five Apis mellifera colonies; one sample from the central frame containing brood and honey, while the other from the frame filled with honey from peripheral side of each colony at every sampling interval. Honey samples were taken by uncapping the cells with knife and extracting the honey with a syringe of 12 mL capacity. Twenty samples of beeswax (one sample from peripheral frames of each colony) were collected from five selected beehives. Two solvents n-hexane and dichloromethane (4:6, v/v) were used for making extracts of honey and beeswax samples, respectively which were further concentrated by rotary evaporation. All the samples were purified and detected at a wavelength of 267 nm. From the HPLC residual analysis, all the honey samples were found free of acaricide flumethrin; as it could not be detected from any of the 40 samples. However, all the beeswax samples were positive for flumethrin residues. The highest level of flumethrin (0.0759 mg/kg) was detected from the samples of beeswax samples taken from the bee colonies after 30 days of flumethrin application. Despite of the fact that flumethrin residues were detected from all the test beeswax samples, however no one was noticed to exceed the tolerance levels established by Environmental Protection Agency (EPA) and European Commission guidelines. Present method of flumethrin detection through HPLC also proved to be a promising alternative of Gas Chromatography with high sensitivity and can be used as an appropriate method to determine flumethrin residues from honey and beeswax.

Evaluation of Physico Chemical Qualities of Beeswax in Selected Districts of Kafa Zone, Southern Nations Nationalities and Peoples Region (SNNPR), Ethiopia

The study was conducted in selected districts of Kafa Zone of Southern Nations Nationalities and Peoples Region (SNNPR), Ethiopia with an intention to identify the physico chemical quality status of beeswax produced in the areas. Three districts; Chena, Gimbo and Gesha and three peasant associations (PAs) from each district were purposively selected based on their potentialities for production and marketing of honey and beeswax. Before collecting sample beeswaxes, a preliminary survey was conducted to identify the major actors taking part in production, processing and marketing of beeswax. Accordingly, beeswax samples were collected from fresh combs, old combs, local mead houses and cooperatives for detail quality analysis. The Mean + SE value for melting point, refractive index, ash content, total volatile matter, acid value, Saponification value, ester value, and ester to acid ratio of beeswax of the area is found to be 62.35 + 0.15, 1.4416 + 0.0001, 0.0857 + 0.0144, 0.4156 + 0.0385,22.689 + 0.3793,94.7540 + 0.8617,72.065 + 0.4766,3.201 + 0.0625 respectively. All the samples passed for paraffin and other waxes, and fats and fatty acid tests. The result shows no significant variation among studied districts in all parameters. However, significantly higher value of refractive index, total volatile matters and ash contents were recorded in samples from local mead houses at p<0.05. This might be due to poor handling practices and an intentional addition of foreign materials such as salt for preserving beeswax from wax moths’ attacks and ‘kocho’ for increasing its volume which are commonly practiced by some individuals of ‘tej’ houses. The overall results revealed that all the beeswax samples collected from various sources are found within the acceptable ranges of national and international standards. Keywords: beeswax, physico-chemical qualities, Kafa zone DOI : 10.7176/CMR/12-1-01 Publication date: January 31 st 2020

Distribution of chemical residues in the beehive compartments and their transfer to the honeybee brood

Honeybee (Apis mellifera) is one of the most important crop and wild plant pollinators, playing an essential role in the agricultural production and the natural ecosystems. However, the number of honeybee colonies is decreasing alarmingly, which has motivated extensive research on the factors affecting their development and survival in some regions. Honeybees' exposure to pesticides and other chemicals has been identified as one of the causes of their decline. The present study evaluates the distribution of plant protection products, veterinary treatments and environmental contaminants inside the beehive, their persistence and their migration to the bee brood. During the five-month sampling period, only amitraz was applied to the colonies. Samples of beeswax, beebread (processed pollen) and bee brood were extracted and analyzed using GC-MS/MS and LC-MS/MS with a multiresidue method. The results showed the presence of 31 chemical residues in the samples. The highest concentrations of residues were detected in the beeswax and corresponded to amitraz (expressed as the sum of DMF and DMPF), coumaphos and tau-fluvalinate, with total concentrations of up to 16,858, 7102 and 1775 μg kg−1, respectively. These and other veterinary treatments were found to accumulate in the beeswax and migrate to other beehive matrices such as beebread and bee brood. Plant protection products used in agriculture were also found in the beehive matrices, especially in the beebread. Five different chemical residues (acrinathrin, amitraz, coumaphos, cypermethrin and tau-fluvalinate) were found in bee brood samples at concentration levels ranging from 1 to 167 μg kg−1. These findings reveal that bee brood reared in field conditions is in fact exposed to plant protection products and veterinary residues through direct contact with contaminated wax and via beebread although they had not been applied to the beehive.

Trace Element Content of Polyfloral Honey and Beeswax from the Vicinity of Non-Ferrous Metal Plant

Bees and their products occupy an important place in the trophic chain “toxicant-soil-plant-bee-bee product-man”, so it has been suggested that bees and bee products should be considered as not a costly method for monitoring of the environmental pollution. Aim of the present study was to analyse trace element content of polyfloral honey and beeswax, produced in three beehives, situated in the vicinity of non-ferrous metal plant and to assess the hazardous risk at consummation. For the purposes of the study, three beehives in this area have been chosen, as follows: BH1 – located at distance of 3.8 km from the smelter, SW; BH2 – at 4 km distance, NE; BH3 – at 4.6 km distance, SE. Content of Al, Cr, Co, Cu, Fe, Mn, Ni, Pb, Sr and Zn was determined by ICP-MS. Data revealed approximately low content of heavy metals and toxic elements. Beeswax samples were found to contain significantly more Cr, Cu, Fe, Pb and Zn, in comparison with the honey samples (p&lt;0.05). On the basis of the trace elements content in bee products, the hazardous risk at consummation could be arranged in the following descending order: BH2 &gt; BH1 &gt; BH3.