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This work aims to encapsulate anthocyanins and phenolic compounds extracted from a native Brazilian fruit peel - jabuticaba (Plinia cauliflora (Mart.) Kausel) and propolis from Tubuna (Scaptotrigona bipunctata) stingless bees, with great potential benefits for human health. The alginate encapsulation was conducted by the ionotropic gelation through the dripping into the CaCl 2 solution. Both raw extracts were characterized by TPC - total phenolic content (Folin-Ciocalteu), AA -antioxidant activity (DPPH and ABTS assays), and TMAC - total monomeric anthocyanin concentration (pH differential method); as well as their resultant mixture (2:1 jabuticaba/propolis). The obtained beads presented highly efficient encapsulation of total polyphenols (~98%) and monomeric anthocyanins (~89%), with spherical morphology and smooth surface obtaining a mean diameter between 200 and 250 μm. In vitro release study showed that JPE/alginate beads were completely disintegrated at pH 7.4 (intestinal pH), but they were resistant to gastric pH (1.2) presenting a slow release of about 40% in 240 min. This is the first report that encapsulates the mixture of jabuticaba and propolis extracts and may contribute to the utilization of a great source of bioactive compounds besides the potential pigment of anthocyanins, an alternative to natural and healthy food/beverage colorants., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Glycosylation is a promising strategy for modulating the physicochemical properties of peptides. However, the influence of glycosylation on the biological activities of peptides remains unknown. Here, we chose the bee venom peptide HYL as a model peptide and 12 different monosaccharides as model sugars to study the effects of glycosylation site, number, and monosaccharide structure on the biochemical properties, activities, and cellular selectivities of HYL derivatives. Some analogues of HYL showed improvement not only in cell selectivity and proteolytic stability but also in antitumor and antimicrobial activity. Moreover, we found that the helicity of glycopeptides can affect its antitumor activity and proteolytic stability, and the α-linked d-monosaccharides can effectively improve the antitumor activity of HYL. Therefore, it is possible to design peptides with improved properties by varying the number, structure, and position of monosaccharides. What's more, the glycopeptides HYL-31 and HYL-33 show a promising prospect for antitumor and antimicrobial drugs development, respectively. In addition, we found that the d-lysine substitution strategy can significantly improve the proteolytic stability of HYL. Our new approach provides a reference or guidance for the research of novel antitumor and antimicrobial peptide drugs.

Nosema ceranae is an emerging and invasive gut pathogen in Apis mellifera and is considered as a factor contributing to the decline of honeybee populations. Here, we used a combined LC-MS and NMR approach to reveal the metabolomics changes in the hemolymph of honeybees infected by this obligate intracellular parasite. For metabolic profiling, hemolymph samples were collected from both uninfected and N. ceranae-infected bees at two time points, 2 days and 10 days after the experimental infection of emergent bees. Hemolymph samples were individually analyzed by LC-MS, whereas each NMR spectrum was obtained from a pool of three hemolymphs. Multivariate statistical PLS-DA models clearly showed that the age of bees was the parameter with the strongest effect on the metabolite profiles. Interestingly, a total of 15 biomarkers were accurately identified and were assigned as candidate biomarkers representative of infection alone or combined effect of age and infection. These biomarkers included carbohydrates (α/β glucose, α/β fructose and hexosamine), amino acids (histidine and proline), dipeptides (Glu-Thr, Cys-Cys and γ-Glu-Leu/Ile), metabolites involved in lipid metabolism (choline, glycerophosphocholine and O-phosphorylethanolamine) and a polyamine compound (spermidine). Our study demonstrated that this untargeted metabolomics-based approach may be useful for a better understanding of pathophysiological mechanisms of the honeybee infection by N. ceranae., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Detection and interpretation of chemical cues is essential for Varroa destructor Anderson and Trueman, an important parasite of honey bees (Apis mellifera L.), to complete its life cycle. We collected volatiles from honey bee brood at various developmental stages and screened for V. destructor electrophysiological responses to these with gas chromatography-linked electrotarsal detection. Volatile collections contained several methyl-alkanes that evoked electrophysiological responses from V. destructor. Moreover, odors in honey bee colonies that regulate honey bee colony structure and function were also detected by V. destructor. Collections from mid- to late-stage larvae had detectable levels of low-volatility odors identified as components of the honey bee brood pheromone and branched alkanes likely originating from brood cuticle. Among these, several mid- to heavy-molecular weight compounds elicited high proportional electrophysiological responses by V. destructor relative to their abundance but could not be identified using chemical standards of previously documented honey bee brood odors. We suggest further investigation of these unknown volatiles and future behavioral assays to determine attractiveness/repellency (valence) of those identified through chemical standards.

Olfaction is a key sensory modality for many arthropods and could be used as a tool in pest management through manipulation of pest behavior. Management of Varroa destructor, important parasitic mites of honey bees, could be improved through better understanding of the chemical ecology of this host-parasite relationship. We refined techniques of mounting mites to obtain electrophysiological recordings (electrotarsograms) of their responses to synthetic odor stimuli. Results of 271 electrotarsogram recordings from V. destructor revealed responses to 10 odorants relative to solvent controls. Electrotarsogram responses to methyl palmitate, ethyl palmitate, and 2-heptanol were highest at the lowest stimulus loading (10 ng) we tested, suggesting that V. destructor may have acute sensitivity to low concentrations of some odors. Results suggest that odorant origin (e.g., methyl oleate from honey bee larvae, geraniol from adult honey bee alarm pheromone, and α-terpineol, a plant secondary metabolite) can influence the degree of electrophysiological response. Varroa destructor tended to be more responsive to known attractants and repellents relative to previously unexplored odorants and some repellent terpenes. Electrotarsograms offer the potential for screening odors to determine their importance in V. destructor host detection.

Background and Aim: Bee pollen is recognized to be a source of different nutrients, including minerals. As a food supplement, its quality and safety due to concentrations of essential macro- and microelements, and harmful trace elements has to be verified. Fast and simple element analysis of bee-collected pollen can be regarded as an important part of its quality assurance and control. The present study aimed at developping a new method for determination of selected elements (Ca, Cu, Fe, Mg, Mn, Zn) of bee pollen based on solvent extraction and completely avoiding a high temperature treatment with concentrated reagents. In addition, in vitro gastrointestinal digestion was used to assess bioavailability of elements from this food supplement., Methods: Bee pollen samples were dried and pulverized. Total concentrations of Ca, Cu, Fe, Mg, Mn, and Zn were determined by flame atomic absorption spectrometry (FAAS) in sample solutions obtained by wet digestion (WD) in concentrated HNO 3 or alternatively by solvent extraction (SE) with diluted solutions of HNO 3 . Gastrointestinal digestion was mimicked using simulated solutions of gastric and intestinal juices followed by determination of Ca, Cu, Fe, Mg, Mn and Zn concentrations in the bioaccessible fraction by FAAS., Results: A new simple and fast method for determination of total concentrations of Ca, Cu, Fe, Mg, Mn, and Zn in bee pollen was developed and validated. The method combined room temperature, two-hour SE with 0.5 mol L -1 HNO 3 with FAAS measurements versus simple standard solutions. It provided precision within 1-5 % and trueness better than 8%, and was shown to be suitable for fast analysis of different polyfloral bee pollens. In vitro gastrointestinal digestion revealed that elements were well (70-85 % for Ca, Mg) and fairly (27-43 % for Cu, Fe, Mn, and Zn) bioaccessible from bee pollen. By pouring with water and swelling overnight, bioaccessibility of studied elements from such prepared bee pollen was increased on average by less than 15 % (Mn), 20 % (Ca, Cu, Fe, Zn) or 30 % (Mn)., Conclusions: Avoiding long-lasting, high-temperature wet digestion with concentrated reagents, the proposed sample treatment along with FAAS provided precise and true results of total concentrations of Ca, Cu, Fe, Mg, Mn, and Zn in bee pollen. The method was simple and fast, and enabled to analyze a higher number of samples. Simulated gastrointestinal digestion of bee pollen have shown for the first time that Ca and Mg are the most bioaccessible from this bee product. Bioaccessibility of Cu, Fe, Mg, and Zn from bee pollen are close to or lower than 40 %., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier GmbH. All rights reserved.)

This study was aimed to investigate the anti-leishmanial effects of bee products (honey and propolis) by using the causative agent of cutaneous leishmaniasis Leishmania tropica promastigotes, in in vitro culture. In vitro anti-leishmanial efficacy of honey (pine, flower and chestnut) and propolis used in the study were evaluated using the microdilution method. Honey, which is a bee product, was dissolved with RPMI medium containing fetal calf serum (FCS) and diluted in the same medium, and serial dilutions were prepared in concentrations between 62.5-1000 mg/ml. Propolis, on the other hand, was dissolved with ethyl alcohol and only 2.5 μl was used from all these concentrations since the alcohol content was more than 50% in these concentrations prepared and we thought that this rate would negatively effect the parasite development. Then, RPMI containing FCS was diluted in the medium and serial dilutions were prepared at concentrations between 50-800 μg/ml. To the dilutions prepared, the promastigot suspension was added so that their final concentrations in the wells were 1 x 106 promastigot/ml and then the medium was incubated for 24 and 48 hours in 26°C. After the incubation, promastigotes were determined microscopically for morphology, mobility and live parasite density, and cell viability was determined by MTS method and 50% inhibitor concentrations (IC50) were compared with control groups. Anti-leishmanial activity of propolis (50, 100, 200, 400 and 800 μg/ml) and honey (62.5, 125, 250, 500 and 1000 mg/ml) on promastigotes was evaluated in vitro. In microscopic examinations, pine honey showed anti-leishmanial activity starting from 62.5 mg/ml, flower honey 250 mg/ml, and chestnut honey 125 mg/ml, and pine honey was more effective on promastigotes (p< 0.05), and propolis was effective from 100 μg/ml concentration. It has been determined that very low concentrations of propolis caused changes in the morphological structure of the parasites and were more effective than the other bee products. The prevention of cell proliferation and decreasing of the IC50 values according with the time of pine honey (IC50= 109.28 mg/ml), flower honey (IC50= 248.07 mg/ml), chestnut honey (IC50= 147.65 mg/ml) and propolis (IC50= 82.98 μg/ml) applied on L.tropica promastigot cell culture was determined by MTS method. In this study, it was found that various concentrations of pine, flower, chestnut honey and propolis showed anti-leishmanial activity on L. tropica promastigotes. It has been observed that pine honey is more effective on promastigotes after 48 hours of incubation period, and propolis is more effective in both morphology and cell inhibition of the parasites even at very low concentrations. It is believed that these data can be used as an alternative treatment method against cutaneous leishmaniasis infections and further studies are required.

To determine how males of the large carpenter bee, Xylocopa appendiculata, maximize access to females while minimizing energy cost and acquiring energy for territorial flights, we investigated the times of territorial flights by males and foraging by males and females. Males were present continuously in territories from 8:00 to 12:00. They approached, chased, and excluded conspecific males from their territories. In the laboratory, males showed higher locomotor and flight activities in the morning and lower activities in the afternoon. Both males and females visited flowers from 8:00 to 16:00, but the most frequent visits were earlier in females (10:00-12:00) than in males (12:00-13:00). Relative body weights in territorial males often increased. These results indicate that the males time their territorial flights to maximize contact with females and obtain nectar as fuel between and after the territorial flights. The time-related territorial flight in males might be based on a time-keeping system in the brain. Brain levels of serotonin and its precursor tryptophan were significantly higher in males collected at 16:00 than at 11:00, suggesting a relation between time-related territorial flight and serotonin synthesis in the brain.

20( S )-Hydroxyvitamin D3 (20(OH)D3) is an endogenous metabolite produced by the action of CYP11A1 on the side chain of vitamin D3 (D3). 20(OH)D3 can be further hydroxylated by CYP11A1, CYP27A1, CYP24A1 and/or CYP27B1 to several hydroxyderivatives. CYP11A1 also hydroxylates D3 to 22-monohydroxyvitamin D3 (22(OH)D3), which is detectable in the epidermis. 20-Hydroxy-7-dehydrocholesterol (20(OH)-7DHC) has been detected in the human epidermis and can be phototransformed into 20(OH)D3 following the absorption of ultraviolet B (UVB) energy by the B-ring. 20(OH)D3 and its hydroxyderivatives have anti-inflammatory, pro-differentiation and anti-proliferative effects, comparable to 1,25-dihydroxyvitamin D3 (1,25(OH) 2 D3). Since cytochromes P450 with 20- or 25-hydroxylase activity are found in insects participating in ecdysone synthesis from 7-dehydrocholesterol (7DHC), we tested whether D3-hydroxyderivatives are present in honey, implying their production in bees. Honey was collected during summer in the Birmingham area of Alabama or purchased commercially and extracted and analyzed using LC-MS. We detected a clear peak of m / z = 423.324 [M + Na] + for 20(OH)D3 corresponding to a concentration in honey of 256 ng/g. We also detected peaks of m / z = 383.331 [M + H - H 2 O] + for 20(OH)-7DHC and 25(OH)D3 with retention times corresponding to the standards. We further detected species with m/z = 407.329 [M + Na] + corresponding to the RT of 7DHC, D3 and lumisterol3 (L3). Similarly, peaks with m/z = 399.326 [M + H - H 2 O] + were detected at the RT of 1,25(OH) 2 D3 and 1,20-dihydroxyvitamin D3 (1,20(OH) 2 D3). Species corresponding to 20-monohydroxylumisterol3 (20(OH)L3), 22-monohydroxyvitamin D3 (22(OH)D3), 20,23-dihydroxyvitamin D3 (20,23(OH) 2 D3), 20,24/25/26-dihydroxyvitamin D3 (20,24/25/26(OH) 2 D3) and 1,20,23/24/25/26-trihydroxyvitamin D3 (1,20,23/24/25/26(OH) 3 D3) were not detectable above the background. In conclusion, the presence of 7DHC and D3 and of species corresponding to 20(OH)-7DHC, 20(OH)D3, 1,20(OH) 2 D3, 25(OH)D3 and 1,25(OH) 2 D3 in honey implies their production in bees, although the precise biochemistry and photochemistry of these processes remain to be defined.

Honey bees play a crucial role in the nature by pollinating wild flowers. Over the past years, there has been an increasing concern regarding the honey bee colony decline. Pesticides or environmental effects targeting the biochemistry of insect chitin and cuticle coating may be in part responsible for honey bee pathologies. We here propose the use of electron paramagnetic resonance imaging (EPRI) as a tool to image the melanin-chitin complexes as part of the exoskeleton of the honey bee. EPRI at 9.65 GHz was applied on intact freeze-dried bees. The imaging data were collected on the melanin peak. High-resolution images revealed that this compound is extensively distributed in the periphery of the animal, data consistent with the localization in the cuticle of the bee. While EPR of melanin has been so far explored in the context of melanoma characterization, it may offer new opportunities in research on honey bees and other insects.

Honey bees (Apis mellifera) forage by using their sense of smell and returning to floral odours that they have previously learned to associate with high-quality food rewards. Foraging bees communicate with other bees in the hive about food sources by exchanging chemical and locational information. It is well established that bees transfer non-volatile information regarding taste and quality of nectar via trophallaxis and communicate location information via directional dances. But to our knowledge, volatiles carried by returning forager bees on their bodies has not been explored as another source of chemical information. We investigated the cuticular-adsorbed odours of bees when foraging on three different crops and compared their odours with the crops' flower headspace. We found that cuticular extracts were in majority correlated with the flower headspace where bees were foraging, specific to the crop and field. Our results support the hypothesis that the scent of returning forager bees can be communicated to hivemates and is associated with information about current floral resources. Some of the floral volatiles that we identified in bee extracts had been previously found to be key compounds learned from the crop, thus supporting a mechanism for the selection of decisive compounds.

Bees are important pollinators whose population has declined due to several factors, including infections by parasites and pathogens. Resource sharing may play a role in the dispersal dynamics of pathogens among bees. This study evaluated the occurrence of viruses (DWV, BQCV, ABPV, IAPV, KBV, and CBPV) and microsporidia (Nosema ceranae and Nosema apis) that infect Apis mellifera, as well as pesticide residues in the stingless bees Nannotrigona testaceicornis, Tetragonisca angustula, and Tetragona elongata sharing the same foraging area with A. mellifera. Stingless bees were obtained from 10 nests (two of N. testaceicornis, five of T. angustula, and three of T. elongata) which were kept in the field for 1 year and analyzed for the occurrence of pathogens. Spores of N. ceranae were detected in stingless bees but were not found in their midgut, which indicates that these bees are not affected, but may be vectors of the microsporidium. Viruses were found in 23.4% of stingless bees samples. APBV was the most prevalent virus (10.8%) followed by DWV and BQCV (both in 5.1% of samples). We detected glyphosate and its metabolites in small amounts in all samples. The highest occurrence of N. ceranae spores and viruses was found in autumn-winter and may be related to both the higher frequency of bee defecation into the colony and the low food resources available in the field, which increases the sharing of plant species among the stingless bees and honey bees. This study shows the simultaneous occurrence of viruses and spores of the microsporidium N. ceranae in asymptomatic stingless bees, which suggest that these bees may be vectors of pathogens.

Here, we present a rational approach that enhances the membrane selectivity of a prolific pore-forming peptide, melittin, based on experimental observations that the cationic polymer, ε-polylysine, disrupts bacterial membranes with greater affinity over mammalian cells when compared to poly-l-lysine and poly-d-lysine. We systematically replaced three α-lysine residues in melittin with ε-lysine residues and identified key residues that are important for cytotoxicity. We then assessed the antimicrobial properties of the modified peptides which carry two or three ε-lysyl residues. Two modified melittin peptides displayed rapid bactericidal properties against antibiotic-resistant strains, low innate resistance development by pathogenic bacteria, remained nonimmunogenic for T lymphocytes, and increased bioavailability in tear fluids. In proof-of-concept in vivo experiments, one of the peptides was noncytotoxic for ocular surfaces and had comparable antimicrobial efficacy to that of fluoroquinolone antibiotics. The results uncover a simple and potential strategy that can enhance the membrane selectivity of cytolytic peptides by ε-lysylation.

Peptidoglycan recognition proteins (PGRPs) play an important role in the defense against invading microbes via the recognition of the immunogenic substance peptidoglycan (PGN). Bees possess fewer PGRPs than Drosophila melanogaster and Anopheles gambiae but retain two important immune pathways, the Toll pathway and the Imd pathway, which can be triggered by the recognition of Dap-type PGN by PGRP-LCx with the assistance of PGRP-LCa in Drosophila . There are three isoforms of PGRP-LC including PGRP-LCx, PGRP-LCa and PGRP-LCy in Drosophila . Our previous study showed that a single PGRP-LC exists in bumblebees. In this present study, we prove that the bumblebee Bombus lantschouensis PGRP-LC (Bl-PGRP-LC) can respond to an infection with Gram-negative bacterium Escherichia coli through binding to the Dap-type PGNs directly, and that E. coli infection induces the quick and strong upregulation of PGRP-LC , abaecin and defensin . Moreover, the Bl-PGRP-LC exhibits a very strong affinity for the Dap-type PGN, much stronger than the affinity exhibited by the PGRP-LC from the more eusocial honeybee Apis mellifera (Am-PGRP-LC). In addition, mutagenesis experiments showed that the residue His 390 is the anchor residue for the binding to the Dap-type PGN and forms a hydrogen bond with MurNAc rather than meso-Dap, which interacts with the anchor residue Arg 413 of PGRP-LCx in Drosophila . Therefore, bumblebee PGRP-LC possesses exclusive characteristics for the immune response among insect PGRPs.

Eusociality is characterized by the reproductive division of labor between two castes: fertile queens and largely sterile workers. Queen pheromones are known to influence worker behavior and reproductive physiology and are therefore key components in regulating complex eusocial behavior [1]. Recent studies indicate that cuticular hydrocarbons (CHCs) act as queen pheromones in various eusocial hymenopteran species [2-8]. However, almost all species investigated to date are highly eusocial and do not include extant transitory stages from solitary to eusocial behavior [9]. Indeed, primitively eusocial species, which largely lack morphologically distinct castes, are thought to control worker reproduction through the physical aggression of the queen rather than via pheromones [10-12]. Halictid or sweat bees exhibit a high variability of eusociality including solitary and facultatively eusocial species [9, 13-16]. However, the mechanisms controlling worker reproduction in these transitory species are unknown. The results of a recent correlative study based on caste-specific chemical profiles in various halictid bees of different social levels have revealed an overproduction of macrocyclic lactones in queens compared with workers [17]. Using chemical analyses and behavioral experiments in which we simulated below-ground nests of the primitively eusocial sweat bee Lasioglossum malachurum, we identified a queen pheromone and found that macrocyclic lactones, not CHCs, influence worker behavior and decrease ovarian activation in this species. Our data suggest that the evolution of queen pheromones is more complex than previously inferred from highly eusocial species and shed new light on the complexity of the evolution of queen pheromones., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Hydrogen has the potential to play an important role in decarbonising our energy systems. Crucial to achieving this is the ability to produce clean sources of hydrogen using renewable energy sources. Currently platinum is commonly used as a hydrogen evolution catalyst, however, the scarcity and expense of platinum is driving the need to develop non-platinum-based catalysts. Here we report a protein-based hydrogen evolution catalyst based on a recombinant silk protein from honeybees and a metal macrocycle, cobalt protoporphyrin (CoPPIX). We enhanced the hydrogen evolution activity three fold compared to the unmodified silk protein by varying the coordinating ligands to the metal centre. Finally, to demonstrate the use of our biological catalyst, we built a proton exchange membrane (PEM) water electrolysis cell using CoPPIX-silk as the hydrogen evolution catalyst that is able to produce hydrogen with a 98% Faradaic efficiency. This represents an exciting advance towards allowing protein-based catalysts to be used in electrolysis cells.

Tertiapin (TPN) is a 21 amino acid venom peptide from Apis mellifera that inhibits certain members of the inward rectifier potassium (Kir) channel family at a nanomolar affinity with limited specificity. Structure-based computational simulations predict that TPN behaves as a pore blocker; however, the molecular determinants mediating block of neuronal Kir3 channels have been inconclusive and unvalidated. Here, using molecular docking and molecular dynamics (MD) simulations with 'potential of mean force' (PMF) calculations, we investigated the energetically most favored interaction of TPN with several Kir3.x channel structures. The resulting binding model for Kir3.2-TPN complexes was then tested by targeted mutagenesis of the predicted contact sites, and their impact on the functional channel block was measured electrophysiologically. Together, our findings indicate that a high-affinity TPN block of Kir3.2 channels involves a pore-inserting lysine side chain requiring (1) hydrophobic interactions at a phenylalanine ring surrounding the channel pore and (2) electrostatic interactions with two adjacent Kir3.2 turret regions. Together, these interactions collectively stabilize high-affinity toxin binding to the Kir3.2 outer vestibule, which orients the ε-amino group of TPN-K21 to occupy the outermost K + binding site of the selectivity filter. The structural determinants for the TPN block described here also revealed a favored subunit arrangement for assembled Kir3.x heteromeric channels, in addition to a multimodal binding capacity of TPN variants consistent with the functional dyad model for polybasic peptide pore blockers. These novel findings will aid efforts in re-engineering the TPN pharmacophore to develop peptide variants having unique and distinct Kir channel blocking properties.

The use of honeybee venom in traditional medicine is increasing due to its unexpected beneficial effects in the treatment of diseases. In this study, a simple and environmentally friendly sample preparation procedure was developed to quantify five biogenic amines-histamine, 5-hydroxytryptamine, dopamine, adrenaline, and noradrenaline-in honeybee venom using high-performance liquid chromatography tandem mass spectrometry. The instrument and sample preparation method were optimized to achieve stable, sensitive, and accurate quantification of the five biogenic amines. The peak purities of five biogenic amines in bee venom were examined using a diode array detector to ensure that endogenous impurities will not interfere with biogenic amines during the chromatographic separation procedure. The correlation coefficient of each compound was higher than 0.998 in the range of 0.5-1000 ng/mL. The limits of detection and quantification of the developed method ranged between 0.09 and 0.17, and 0.3 and 0.59 μg/g, respectively. The average recoveries of spiked biogenic amines with different concentrations were higher than 70.95%, and the intra- and intermediate-day precisions were lower than 7.51% and 10.17%, respectively. The carry-over between each injection and the stability of the target analytes were also evaluated to ensure the effectiveness of this method. The data obtained are presented in various formats, including boxplot, heat map, and principal component analysis diagram, to visualize the differences in the biogenic amine contents of the honeybee venoms from different subspecies. This method hopes to provide the opportunity to distinguish the bee venom produced by different subspecies., (© 2019 John Wiley & Sons, Ltd.)

Royal jelly (RJ) is a yellowish-white and acidic secretion of hypopharyngeal and mandibular glands of nurse bees used to feed young worker larvae during the first three days and the entire life of queen bees. RJ is one of the most appreciated and valued natural product which has been mainly used in traditional medicines, health foods, and cosmetics for a long time in different parts of the world. It is also the most studied bee product, aimed at unravelling its bioactivities, such as antimicrobial, antioxidant, anti-aging, immunomodulatory, and general tonic action against laboratory animals, microbial organisms, farm animals, and clinical trials. It is commonly used to supplement various diseases, including cancer, diabetes, cardiovascular, and Alzheimer's disease. Here, we highlight the recent research advances on the main bioactive compounds of RJ, such as proteins, peptides, fatty acids, and phenolics, for a comprehensive understanding of the biochemistry, biological, and pharmaceutical responses to human health promotion and life benefits. This is potentially important to gain novel insight into the biological and pharmaceutical properties of RJ.

Propolis is a resin that honeybees produce by mixing saliva and beeswax with exudate gathered from botanical sources. The present in vitro study investigated the potential use of propolis as a multitarget therapeutic product and the physicochemical properties, chemical composition, and immunomodulatory, antioxidant, antibacterial, and anticancer properties of a propolis extract from the northern Morocco region (PNM). Pinocembrin, chrysin, and quercetin were the main phenolic compounds of PNM as measured in HPLC. The PNM showed significant inhibitory effects against all tested Gram-positive and Gram-negative strains and showed high antioxidant activities by scavenging free radicals with IC50 (DPPH = 0.02, ABTS = 0.04, and FRAP = 0.04 mg/ml). In addition, PNM induced a dose-dependent cytostatic effect in MCF-7, HCT, and THP-1 cell lines at noncytotoxic concentrations with IC50 values of 479.22, 108.88, and 50.54  μ g/ml, respectively. The production of tumor necrosis factor- α (TNF- α ) and interleukin-6 (IL-6) was decreased in a dose-dependent manner in LPS-stimulated human peripheral blood mononuclear cells (PBMNCs), whereas the production of the anti-inflammatory interleukin-10 (IL-10) was increased in a dose-dependent manner reaching 15-fold compared to the levels measured in untreated PBMNCs. Overall, the results showed that the traditionally known multitarget therapeutic properties of the PNM seem to be mediated, at least in part, through cytostatic, antibacterial, and immunomodulatory effects., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2019 Soumaya Touzani et al.)

Honeybees communicate with members of their intra-species via pheromones. The volatile pheromones, β-ocimene and allo-ocimene, are the primary signals of larvae to beg for the care from the nurses. Of the odorant binding proteins (OBPs)/chemosensory proteins (CSPs), CSP4 has the best affinity with β-ocimene and allo-ocimene. To reveal the binding mechanism of CPS4 with them, fluorescent quenching, UV absorption spectra, circular dichroism (CD) spectra, isothermal titration calorimetry (ITC), molecular docking, molecular dynamic (MD) simulation, and site-directed mutagenesis were applied. The quenching constant Ksv decreased with temperature increase, and the interaction distance was 2.73 nm and 2.43 nm (<10 nm), indicating that β-ocimene and allo-ocimene could form stable complexes with CSP4. The observed △H < 0 and △S > 0 of thermodynamics suggest the main driving forces are electrostatic or hydrophobic force. All above thermodynamics findings are in line with the results of ITC experiments. Furthermore, molecular docking, MD simulation and site-directed mutagenesis indicate the binding cavities are located at cavity 1 in C-terminal of CSP4, where Tyr98 and Asp67 are vital amino acids in maintaining the stable form of protein and larval pheromones, and electrostatic energies are the main driving forces. Our findings gain novel insight into the binding mechanism of chemosensory protein with volatile larval pheromones and are important for understanding olfactory interaction of honeybees., (Copyright © 2019. Published by Elsevier B.V.)

Salmonella is a major global food-borne pathogen. One of the main concerns related to Salmonella and other food-borne pathogens is their capacity to acquire antimicrobial resistance and produce biofilms. Due to the increased resistance to common antimicrobials used to treat livestock animals and human infections, the discovery of new antimicrobial substances is one of the main challenges in microbiological research. An additional challenge is the development of new methods and substances to inhibit and destruct biofilms. We determined the antimicrobial and antibiofilm activities of apitoxin in 16 Salmonella strains isolated from poultry. In addition, the effect of apitoxin on Salmonella motility and the expression of biofilm- and virulence-related genes was evaluated. The minimum inhibitory concentrations (MIC) of apitoxin ranged from 1,024-256 μg/mL, with 512 μg/mL being the most common. Sub-inhibitory concentrations of apitoxin significantly reduced biofilm formation in 14 of the 16 Salmonella strains tested, with significant increases in motility. MIC concentrations of apitoxin destroyed the pre-formed biofilm by 27.66-68.22% (47.00% ± 10.91). The expression of biofilm- and virulence-related genes and small RNAs was differentially regulated according to the strain and the presence of apitoxin. The transcription of the small RNAs dsrA and csrB, related to antimicrobial resistance, was upregulated in the presence of apitoxin. We suggest that apitoxin is a potential antimicrobial substance that could be used in combination with other substances to develop new drugs and sanitizers against food-borne pathogens., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

During foraging, bees are exposed to sublethal doses of insecticides, which can cause morphological changes to various organs, such as the midgut, Malpighian tubules, and mushroon body. Thus, the purpose of this study was to establish a scoring system to evaluate these alterations based on the damage caused and its reversibility. Therefore, a grade 1 score indicates a minimal and easily reversible lesion, increased apocrine secretion, increased cell elimination into the lumen, and a larger quantity of spherocrystals; grade 2 was assigned to moderate and typically reversible injuries, such as changes in the brush border, vacuolation/loss of cytoplasmic material, presence/height of the brush border, and cell swelling; and grade 3 was assigned to serious and irreversible, loss of cell nests of regenerative cells, pyknosis, and loss of contact between Kenyon cells. In addition, frequency values were assigned since the alterations can occur at different frequencies according to the insecticide and the bees exposed; the frequency ranges from 0 to 6, with 0 representing the absence of an alteration and 6 representing a high-frequency occurrence. Based on the analyses, we conclude that each change causes morphological damage, which may or may not be irreversible and could affect the health of the colony., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The current research aimed to quantify melittin (MEL) in Iranian honey bee (Apis mellifera meda) venom. To this end, a liquid chromatography-electrospray ionization-ion trap tandem mass spectrometry (LC-ESI-IT-MS/MS) approach was employed. Melittin is the main toxic peptide of honey bee venom with various biological and pharmacological activities. It was extracted with pure water from the bee venom samples. The analyses were performed on XBridge BEH300 C4 column using a gradient method with the mobile phase consisting of ultrapure water and acetonitrile (containing 0.1% formic acid). Signals of the melittin were recorded with the selected reaction monitoring (SRM) mode, which is a quantitative approach capable of quantifying analyte peptides with high sensitivity and. The mass spectrum of MEL was obtained in the positive ion mode and the quantification analysis was performed using precursor to product ion transition of m/z 570.2/669.9. This method demonstrated good linearity (R2˃0.997) in the range of 1-100 &micro;g mL-1, with a limit of quantification (LOQ) of 1.0 &micro;g mL-1. The content of MEL in Iranian honey bee venom accounts for 43&ndash;55% of total dry weight. This method can be used to evaluate the quality and authenticity of bee venom samples for different therapeutic applications of MEL., (Copyright © 2019, Archives of Razi Institute. Published by Kowsar.)

Nanotechnology has the potential to overcome the challenges of sustainable agriculture, and nanopesticides can control agricultural pests and increase farm productivity with little environmental impact. However, it is important to evaluate their toxicity on non-target organisms, such as honeybees (Apis mellifera) that forage on crops. The aims of this study were to develop a nanopesticide that was based on solid lipid nanoparticles (SLNs) loaded with pyrethrum extract (PYR) and evaluate its physicochemical properties and short-term toxicity on a non-target organism (honeybee). SLN + PYR was physicochemically stable after 120 days. SLN + PYR had a final diameter of 260.8 ± 3.7 nm and a polydispersion index of 0.15 ± 0.02 nm, in comparison with SLN alone that had a diameter of 406.7 ± 6.7 nm and a polydispersion index of 0.39 ± 0.12 nm. SLN + PYR had an encapsulation efficiency of 99%. The survival analysis of honeybees indicated that PYR 10ng presented shorter longevity than those in the control group (P ≤ 0.01). Empty nanoparticles and PYR 10ng caused morphological alterations in the bees' midguts, whereas pyrethrum-loaded nanoparticles had no significant effect on digestive cells, so are considered safer, at least in the short term, for honeybees. These results are important in understanding the effects of nanopesticides on beneficial insects and may decrease the environmental impacts of pesticides., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Background: Pesticides have often been linked to honey bee colony losses, which occur mainly over winter. In this study, we investigated residues in nine colonies at a model agricultural research site during the period before wintering. Moreover, we applied the acaricide tau-fluvalinate to the colonies via a strip formulation. The pesticide content was determined by UHPLC-QqQ-MS/MS in bees from brood comb initially collected in mid-September immediately prior to the start of tau-fluvalinate treatment and 30 later at the time of tau-fluvalinate strip removal., Results: In addition to commonly analyzed pesticides, we detected two plant growth regulators, chlormequat and metazachlor, in the bee colonies. Whereas thiacloprid, chlormequat and acetamiprid decreased after 30 days and contributed considerably to differences between sample time points, other pesticides appeared to be rather stable. Interestingly, we identified diazinon, which has been banned in the European Union since 2007. The residues of methiocarb sulfoxide and imidacloprid-urea in the absence of their parent compounds indicate historical environmental contamination that can be identified by the detection of residues in a bee colony. tau-Fluvalinate was detected only after the 30-day treatment at an average (± SD) concentration of 1.29 ± 1.93 ng/bee, ranging from 0.06 to 7.13 ng/bee., Conclusion: The multidimensional behavior of pesticides in a bee colony was indicated. Although the research area is used for agriculture, the measured pesticide level was relatively low. The recorded concentrations of tau-fluvalinate should not be dangerous to bees, as the values were ∼ 200-5000-fold lower than the reported median lethal dose (LD 50 ) values. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

Information on compositional, nutritional and functional properties of bee-pollen, as a health-promoting food, is essential for defining its quality. Concerning the nutritional importance of phenolic compounds, the aim of this study was to determine the phenolic profile and antioxidant activity of twenty-four bee-pollen samples collected from different regions of Serbia. High-performance thin-layer chromatographic (HPTLC) fingerprinting was used for profiling of bee-pollen samples according to the botanical type. HPTLC hyphenated with image analysis and a pattern recognition technique confirmed the grouping of samples caused by the specific phenolic composition of pollens of different botanical origin. Flavonoid glycosides in bee-pollen samples were identified by applying ultra-high-performance liquid chromatography (UHPLC) coupled with linear ion trap-Orbitrap mass spectrometry (LTQ Orbitrap MS). Eight out of twenty-seven flavonol glycosides were identified in bee-pollen samples for the first time. All analyzed bee-pollen samples showed a high number of phenolic compounds which may have therapeutic potential.

In the last years, the honeybee population is facing growing threats such as expansion of pathogens, incorrect use of phytosanitary products and environmental contaminants, loss or fragmentation of habitat, invasive species and climate change. The citrus cropping by Integrated Pest Management (IPM) in Spain combines strategies to reduce pest populations preventing environmental problems and reducing levels of damage by using chemicals only when it is strictly necessary. The goal of this study is to develop a simple analytical method to evaluate pesticide residue levels in honeybees and corbicular pollen when honeybees are exposed to plant protection products (PPPs) used in integrated pest management citrus orchards. The proposed method is based in an ultrasound assisted extraction procedure followed by a dispersive solid phase extraction (d-SPE) clean-up with alumina and LC-MS/MS pesticides determination. The method was validated in samples of honeybee and corbicular pollen for 10 pesticides commonly used in citrus orchards under IPM. This procedure was compared with QuEChERS methodologies for these matrices. The developed method was applied to determine pesticides in both matrices in a two -year study in citrus orchards., (Copyright © 2019. Published by Elsevier B.V.)

Queen pheromones effect the reproductive division of labour, a defining feature of eusociality. Reproductive division of labour ensures that one, or a small number of, females are responsible for the majority of reproduction within a colony. Much work on the evolution and function of these pheromones has focussed on Queen Mandibular Pheromone (QMP) which is produced by the Western or European honeybee (Apis mellifera). QMP has phylogenetically broad effects, repressing reproduction in a variety of arthropods, including those distantly related to the honeybee such as the fruit fly Drosophila melanogaster. QMP is highly derived and has little chemical similarity to the majority of hymenopteran queen pheromones which are derived from cuticular hydrocarbons. This raises the question of whether the phylogenetically widespread repression of reproduction by QMP also occurs with more basal saturated hydrocarbon-based queen-pheromones. Using D. melanogaster we show that saturated hydrocarbons are incapable of repressing reproduction, unlike QMP. We also show no interaction between the four saturated hydrocarbons tested or between the saturated hydrocarbons and QMP, implying that there is no conservation in the mechanism of detection or action between these compounds. We propose that the phylogenetically broad reproductive repression seen in response to QMP is not a feature of all queen pheromones, but unique to QMP itself, which has implications for our understanding of how queen pheromones act and evolve., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Stingless bee honey produced by Heterotrigona itama from different botanical origins was characterised and discriminated. Three types of stingless bee honey collected from acacia, gelam, and starfruit nectars were analyzed and compared with Apis mellifera honey. The results showed that stingless bee honey samples from the three different botanical origins were significantly different in terms of their moisture content, pH, free acidity, total soluble solids, colour characteristics, sugar content, amino acid content and antioxidant properties. Stingless bee honey was significantly different from Apis mellifera honey in terms of physicochemical and antioxidant properties. The amino acid content was further used in the chemometrics analysis to evaluate the role of amino acid in discriminating honey according to botanical origin. Partial least squares-discriminant analysis (PLS-DA) revealed that the stingless bee honey was completely distinguishable from Apis mellifera honey. Notably, a clear distinction between the stingless bee honey types was also observed. The specific amino acids involved in the distinction of honey were cysteine for acacia and gelam, phenylalanine and 3-hydroxyproline for starfruit, and proline for Apis mellifera honey. The results showed that all honey samples were successfully classified based on amino acid content.

Honeybees, due to their wide-ranging foraging behaviour, have great potentials for monitoring environmental quality. Therefore, the purpose of this study was to use honeybees as bioindicators for the detection of toxic and essential metals (Cd, Cr, Cu, and Fe) in the environment. Totally, 180 soil, plant (including root and aerial part), honeybees, and honey samples were collected from 12 sites within the main beekeeping and honey production regions in Markazi Province, Iran in 2016. After acid digestion of samples, the metal concentrations were measured by inductively coupled plasma-optical emission spectrometers. The translocation factor (TF), and bioconcentration factor (BCF) of metals were computed. The results showed that among the analyzed bee samples, Cd (mg kg -1 ) was detected in amounts ranging from 0.01 to 2.35, Cr (mg kg -1 ) ranged from 0.02 to 18.10, Cu (mg kg -1 ) ranged from 2.00 to 39.11, and Fe (mg kg -1 ) ranged from 163 to 1695. BCF and TF values obtained showed that the Astragalus gossypinus would have a great accumulation ability for Cd and Cr. The results indicated that honeybees could be used to detect the spatial patterns of metal contaminations in the environment they dwell in.

Pesticide residues in beebread, live and dead honey bees, together with honey bee death rate were monitored from June 2016 to June 2018 in three apiaries, located near agricultural settings and in wildlands. Dead honey bees were only collected and analyzed when significant mortality episodes occurred and pesticide content in beeswax of each experimental apiary was evaluated at the beginning of the study. Samples were extracted by a modified QuEChERS procedure and screened for pesticides residues by liquid chromatography mass spectrometry (LC-MS/MS). Pesticide hazard in the samples was evaluated through the hazard quotient approach (HQ). Beebread was widely contaminated with coumaphos and amitraz degradate 2, 4-dimethylphenylformamide (DMF), miticides detected in 94 and 97% of samples respectively. However, insecticides sprayed during citrus bloom like chlorpyrifos (up to 167 ng g -1 ) and dimethoate (up to 34 ng g -1 ) were the main responsible of the relevant pesticide hazard in this matrix. Pesticide levels in live bees were mostly residual, and pesticide hazard was low. Beeswax of the apiaries, contaminated by miticides, revealed a low pesticide hazard to honey bee colonies. Acute mortality episodes occurred only in the two apiaries located near agricultural settings. Dead bees collected during these episodes revealed high levels (up to 2700 ng g -1 ) of chlorpyrifos, dimethoate, omethoate and imidacloprid. HQ calculated in dead bees exceeded up to 37 times the threshold value considered as elevated hazard to honey bee health., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Bee pollen (BP) is a natural medicine from the hive with various potential health-promoting benefits, but until now there is no study to determine its protective roles in inflammatory bowel disease (IBD). The aim of this study was to reveal the in vitro gastrointestinal protective effects of BP against IBD using molecular and metabolic methods. Dextran sulfate sodium (DSS) challenged Caco-2 cell monolayers were applied to mimic intestinal epithelial cell dysfunctions and metabolic disorders. The pretreatment with BP extract rich in polyphenols ameliorated DSS-induced cell viability losses. It also exerted protective effects against intestinal barrier impairment by strengthening epithelial integrity and tight junction losses induced by DSS. BP up-regulated anti-oxidant (NQO1, Txnrd1, Nrf2) and down-regulated inflammatory (TNF-α and IL-6) mRNA expressions, in accompany with MAPK signaling inhibition. Furthermore, metabolomics analysis based on UPLC-Q-TOF/MS revealed that BP, and DSS treated Caco-2 cells have different metabolomic profiles, with significant changes on key metabolites involved in glycerophospholipid metabolism. Our results showed that BP has great therapeutic potential throughout the early stages of DSS-induced colitis., (Copyright © 2019 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Oral mucositis is one of the most frequent complications after chemotherapy or radiotherapy or a combination of both. There is no standard therapy for its prevention or treatment. Considering that some bee products have been found to be of value in this situation, we decided to analyze the scientific literature on the subject. Scientific publications on bee products were identified by a literature search on Pubmed, Scopus and Google Scholar. There is a lot of evidence regarding the use of honey for oral mucositis due to chemotherapy or radiotherapy or a combination of both. Unfortunately, the quality of several meta-analyses on the topic is very low. There is some evidence on propolis, a little on royal jelly and none whatsoever on pollen and other bee products like apilarnil or bee venom. Bee products such as honey, propolis and royal jelly may be well suited to be integrated into a general concept for the prevention and treatment of oral mucositis which should also include other established concepts like oral care, oral cryotherapy, topical vitamin E and low-level-laser therapy. Bee products could become an integral part in the treatment of chemotherapy, radiotherapy and radio chemotherapy. High-quality meta-analyses and further studies, especially on the combinations of various strategies, are needed.

Apitherapy is an alternate therapy that relies on the usage of honeybee products, most importantly bee venom for the treatment of many human diseases. The venom can be introduced into the human body by manual injection or by direct bee stings. Bee venom contains several active molecules such as peptides and enzymes that have advantageous potential in treating inflammation and central nervous system diseases, such as Parkinson's disease, Alzheimer's disease, and amyotrophic lateral sclerosis. Moreover, bee venom has shown promising benefits against different types of cancer as well as anti-viral activity, even against the challenging human immunodeficiency virus (HIV). Many studies described biological activities of bee venom components and launched preclinical trials to improve the potential use of apitoxin and its constituents as the next generation of drugs. The aim of this review is to summarize the main compounds of bee venom, their primary biological properties, mechanisms of action, and their therapeutic values in alternative therapy strategies.

If tolerated in biological environments, recombinant structural proteins offer the advantage that biological cues dictating cell attachment and material degradation can be modified as required for clinical application using genetic engineering. In this study, we investigate the biological response to materials generated from the recombinant honeybee silk protein, AmelF3, a structural protein that can be produced at high levels by fermentation in Escherichia coli. The protein can be readily purified from E. coli host cell proteins after transgenic production and fabricated into various material formats. When implanted subcutaneously according to International Standard ISO 10993 tests, materials generated from the purified recombinant protein were found to be noncytotoxic, inducing a transient weak immunogenic response and a chronic inflammatory response that resolved over time. While preliminary, this study supports the ongoing development of materials generated from this protein for biomedical applications. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 1763-1770, 2019., (© 2019 Wiley Periodicals, Inc.)

The current study was set to prepare a delivery system based on a mixture of biodegradable polymeric hydrogel (i.e., carrageenan (κ-CAR) and β-cyclodextrin (β-CD)) for encapsulating honey bee propolis extract (HBP). The crosslinking agent; glyoxal was incorporated to increase the insolubility of the HBP-loaded hydrogel to fit the cotton fabric treatments. The synthesized hydrogels were fully characterized to examine their surface and internal morphology and to visualize the chemical interaction between κ-CAR and β-CD via SEM and FTIR techniques. The SEM pictures ensured the existent of many pores within the formed hydrogel. The swelling properties of κ-CAR/β-CD/HBP were also studied in terms of glyoxal and HBP concentrations which displayed water holding with higher mechanical properties. Furthermore, in vitro release assessment of HBP from κ-CAR/β-CD hydrogel was also studied at pH 7.6. Therefore, κ-CAR/β-CD/HBP hydrogel was successfully used as fining compound for the cationized cotton fabrics. The data obtained emphasized that the surface of cationized cotton fabrics was deposited and penetrated with a film of κ-CAR/β-CD/HBP. In addition, the antimicrobial activity of the treated cationized fabric was evaluated against many species of bacteria and fungus producing highly inhibition zone at lower concentrations of HBP. Accordingly, the cationized cotton fabric treated with κ-CAR/β-CD/HBP has a high potential in the wound healing fabrics., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Previous studies have shown that pollen products sold as nutritional supplements and used in apitherapy may contain toxic pyrrolizidine alkaloids (PAs) if bees collect pollen from PA-containing plants, such as Echium vulgare . In this study, the botanical origin of pollen from two observation sites was studied. Despite a high PA content in pollen samples that bees collected during E. vulgare 's flowering period, bees were found to collect relatively few Echium pollen loads. Thus, the monitoring of pollen loads collected at the apiaries is unviable to estimate the risk of PA contamination in pollen or bee bread. In a second step, the stability of PAs in bee bread samples containing PAs at concentrations of 2538 ng/g and 98 ng/g was assessed over a period of five or six months, respectively. No significant PA reduction was observed in bee bread stored at 15 °C, but there were overall PA reductions of 39% and 33% in bee bread stored at 30 °C, reflecting hive conditions. While PA N -oxides decreased over time, other types of PAs remained relatively stable. Monitoring PAs in pollen products remains important to ensure consumer safety and should include echivulgarine (and its N -oxide), the major PA type found in pollen from E. vulgare .

Flor velum yeast growth activators during biological aging are currently unknown. In this sense, this research focuses on the use of bee pollen as a flor velum activator. Bee pollen influence on viable yeast development, surface hydrophobicity, and yeast assimilable nitrogen has already been studied. Additionally, bee pollen effects on the main compounds related to flor yeast metabolism and wine sensory characteristics have been evaluated. "Fino" (Sherry) wine was supplemented with bee pollen using six different doses ranging from 0.1 to 20 g/L. Its addition in a dose equal or greater than 0.25 g/L can be an effective flor velum activator, increasing yeast populations and its buoyancy due to its content of yeast assimilable nitrogen and fatty acids. Except for the 20 g/L dose, pollen did not induce any significant effect on flor velum metabolism, physicochemical parameters, organic acids, major volatile compounds, or glycerol. Sensory analysis showed that low bee pollen doses increase wine's biological aging attributes, obtaining the highest score from the tasting panel. Multiflora bee pollen could be a natural oenological tool to enhance flor velum development and wine sensory qualities. This study confirms association between the bee pollen dose applied and the flor velum growth rate. The addition of bee pollen could help winemakers to accelerate or reimplant flor velum in biologically aged wines.

Propolis produced by the stingless bee Lisotrigona cacciae was studied for the first time. Using different chromatographic procedures, a total of eighteen constituents (phenols and triterpenes) were isolated, among which flavane 1, homoisoflavanes 2-4, and xanthones 5 and 6 were new for propolis. Propolis extract was also characterized by gas chromatography/mass spectrometry and other fifteen constituents were identified. The xanthone α-mangostin (8) demonstrated significant activity against Staphylococcus aureus with MIC and MBC 0.31 μg/ml, followed by 7,4'-dihydroxy-5-methoxy-8-methylflavane (1) with MIC 78 μg/ml and MBC 156 μg/ml. 10,11- Dihydroxydracaenone C (4), a component bearing ortho-hydroxyl groups, was the only compound displaying radical scavenging ability. Triple botanical origin of the sample was defined, consisting of Dracaena cochinchinensis, Cratoxylum cochinchinense and Mangifera indica. D. cochinchinensis is a new resin source of propolis., Competing Interests: The authors have declared that no competing interests exist.

The emergence of antibiotic-resistant bacteria, dubbed superbugs, together with relative stagnation in developing efficient antibiotics has led to enormous health and economic problems, necessitating the need for discovering and developing novel antimicrobial agents. In this respect, animal venoms represent a rich repertoire of pharmacologically active components. As a major component in the venom of European honeybee Apis mellifera, melittin has a great potential in medical applications. In this mini-review, we summarize a multitude of studies on anti-bacterial effects of melittin against planktonic and biofilm-embedded bacteria. Several investigations regarding synergistic effects between melittin and antibiotics were also described. On the whole, the properties of melittin can open up new horizons in a range of biomedical areas, from agriculture to veterinary and clinical microbiology.

The continuous discharge of diverse chemical products in the environment is nowadays of great concern to the whole world as some of them persist in the environment leading to serious diseases. Several sampling techniques have been used for the characterization of this chemical pollution, although biomonitoring using natural samplers has recently become the technique of choice in this field due to its efficiency, specificity, and low cost. In fact, several living organisms known as biomonitors could accumulate the well-known persistent environmental pollutants allowing their monitoring in the environment. In this work, a review on environmental biomonitoring is presented. The main sampling techniques used for monitoring environmental pollutants are first reported, followed by an overview on well-known natural species used as passive samplers and known as biomonitors. These species include conifer needles, lichen, mosses, bees and their byproducts, and snails, and were widely used in recent research as reliable monitors for environmental pollution.

Large social insect colonies exhibit a remarkable ability for recognizing group members via colony-specific cuticular pheromonal signatures. Previous work suggested that in some ant species, colony-specific pheromonal profiles are generated through a mechanism involving the transfer and homogenization of cuticular hydrocarbons (CHCs) across members of the colony. However, how colony-specific chemical profiles are generated in other social insect clades remains mostly unknown. Here we show that in the honey bee ( Apis mellifera ), the colony-specific CHC profile completes its maturation in foragers via a sequence of stereotypic age-dependent quantitative and qualitative chemical transitions, which are driven by environmentally-sensitive intrinsic biosynthetic pathways. Therefore, the CHC profiles of individual honey bees are not likely produced through homogenization and transfer mechanisms, but instead mature in association with age-dependent division of labor. Furthermore, non-nestmate rejection behaviors seem to be contextually restricted to behavioral interactions between entering foragers and guards at the hive entrance., Competing Interests: CV, JK, KM, AH, JL, YB No competing interests declared, (© 2019, Vernier et al.)

Queen mandibular pheromone (QMP) is a potent reproductive signal to which honey bee workers respond by suppressing their ovaries and adopting alloparental roles within the colony. This anti-ovarian effect of QMP on workers can, surprisingly, be induced in other insects, including fruit flies, in which females exposed to synthetic QMP develop smaller ovaries with fewer eggs. In this study, we use the Drosophila melanogaster model to identify the components of synthetic QMP required for the anti-ovarian effect. We found that virgin females respond strongly to 9-oxo-2-decenoic acid and 10-hydroxy-2-decenoic acid (10HDA), suggesting that the decenoic acid components of QMP are essential for the anti-ovarian response. Further, a nuclear factor of activated T-cells reporter system revealed neurones expressing the olfactory receptors Or-56a, Or-49b and Or-98a are activated by QMP in the antenna. In addition, we used olfactory receptor GAL4 drivers and a neuronal activator (a neuronal activating bacterial sodium channel) to test whether the candidate neurones are potential labelled lines for a decenoic acid response. We identified Or-49b as a potential candidate receiver of the 10HDA signal. Finally, the anti-ovarian response to synthetic QMP is not mediated by decreasing the titre of the reproductive hormones ecdysone and juvenile hormone., (© 2018 The Royal Entomological Society.)

In our research soluble proteins from edible grasshopper (S. gregaria) and honey bee brood (A. mellifera) were exposed to defatting, alkaline, and sonication-assisted extractions. New nitrogen-to-protein conversion factors based on amino acid analysis were estimated for both insects: 4.5 for adult grasshopper, and 4.9 and 5.6 for pupae and larvae of honey bee respectively, in contrary to 6.25 commonly used for insects. All fractions were characterized by their composition, yield, color, protein solubility, and functional properties in comparison to whey protein concentrate. Besides an increase in protein content up to 57.5 and 55.2% for grasshopper and honey bee respectively, protein-enriched fractions showed improved foaming and emulsifying abilities. The highest emulsion stability after 120 min was determined for grasshopper powder extracted with sonication (85.5%) and whey proteins (89.8%). The protein-enriched fractions of both insects had significantly higher foaming stability (74.1% for grasshopper fraction after alkaline extraction and 55.5% for sonication-assisted honey bee fraction) than raw and defatted powder. All fractions obtained from honey bee brood showed significantly higher protein heat coagulation than grasshopper and whey proteins. Changes in protein functionality were found related to alteration in protein charge, surface hydrophobicity, and distribution of proteins according to their molecular weight. Therefore, our results showed that S. gregaria and A. mellifera have a potential for future applications for food, feed, or insect-based dietary supplements., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Colonization by toxin-producing bacteria in the gut plays a major role in bowel problems in autistic patients. Prebiotics can inhibit the growth of these pathogenic microbes by nourishing beneficial bacteria, while probiotics--live microorganisms--can balance the gut bacteria; thus, both together can maintain healthy bacteria in the gut. The present study was conducted to find the effect of probiotics and prebiotics in balancing the gut flora in a rodent model of autism linked with a clindamycin-induced altered gut. The effects of probiotics and prebiotics on oxidative stress markers in the brain were also evaluated. Eight groups of hamsters were assigned, with Group I serving as the control; Group II, as the autistic model, was treated with 250 mg propionic acid/kg BW/day for 3 days; Group III was treated with clindamycin 30 mg/kg BW for 3 days; Groups IV and V were treated with bee pollen and propolis (supposed prebiotics) at a dose of 250 mg/kg BW/day for 28 days; Group VI and Group VII were treated with Lactobacillus paracaseii and Protexin® (supposed probiotics) for 28 days; and finally, Group VIII was anorectally transplanted with stool from normal animals for 5 days. Remarkable changes were measured in oxidative stress markers, primarily glutathione and vitamin C, in the brains of hamsters in the propionic acid- and clindamycin-treated group. All probiotic/prebiotic treatments showed ameliorative effects; however, lactobacillus had the strongest effect. We conclude that pro-and prebiotic supplements may be effective to revive healthy digestive system function in autistic patients. The disappointing results of the fecal transplants suggest that further study is needed to develop an appropriate technique.

This study reports an analytical approach by gas chromatography and high-resolution mass spectrometry (HRMS) intended to be used for investigation of non-targeted environmental contaminants in honeybees. The approach involves a generic extraction and analysis with two GC-HRMS systems: time-of-flight and Orbitrap analyzers, GC-TOF-MS, and GC-Orbitrap-MS operated in electron-impact ionization (EI) mode. The workflow for screening of non-targeted contaminants consisted of initial peak detection by deconvolution and matching the first-stage mass spectra EI-MS with a nominal mass spectral library. To gain further confidence in the structural characterization of the contaminants under investigation, molecular formula of representative ions (molecular and fragment ions) was provided for those with an accurate mass scoring (error < 5 ppm). This methology was applied for screening environmental contaminants in 75 samples of adult honeybee. This approach has provided the tentative identification of environmental contaminants belonging to different chemical groups, among them, PAHs, phthalates and synthetic musks. Residues of veterinary treatments used in apiculture were also detected in the honeybee samples., (Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.)

The inspiration for this study was the anxiety of Warsaw beekeepers, who raised the question whether location of hives in large urban agglomerations results in changes in concentrations of xenobiotics, toxic elements, and micronutrients in honey bees. Preliminary studies required elaboration of the research methodology, as the studied object is characterized by a low degree of homogeneity and the method of sample preparation affects obtained results. From many tested approaches, the use of washed and milled abdomens of the bees is recommended. Results obtained for such prepared samples are slightly lower than for whole bees, but their repeatability is higher, which enables easier interpretation of the trends and comparison of different locations. The contents of selected elements (As, Al, Cd, Co, Cr, Cu, Mn, Pb, and Zn) were compared in bees from urban and rural areas. The studies were supported by pesticides analysis. Also, it was checked whether these substances are accumulated on the surface or inside the bee's body. The research indicates the markers of contamination: Al, As, and Cr on the surface and Cd inside the bodies of honey bees. The location of the hives does not influence significantly the content of "toxic," nutrient metals and metalloids in bees (slightly higher levels of As, Al, Pb, and Cd were found in bees from urban areas). In terms of exposure to these elements and pesticides, the large city environment is not harmful for honey bees.

An analytical method based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the first simultaneous determination of ten neonicotinoid insecticides (pymetrozine, dinotefuran, nitenpyram, thiamethoxam, flonicamid, imidacloprid, clothianidin, imidaclothiz, acetamiprid, thiacloprid) and two metabolites (4-trifluoromethylnicotinamide and N-desmethylacetamiprid) in honey and royal-jelly. The quantitative detection was performed by LC-MS/MS on multiple reaction monitoring (MRM) mode under positive-ion electrospray ionization. The isotope dilution internal standard method or external standard method was used for quantitation analysis. The limits of quantification (LOQs), calculated as 10 times the standard deviation, were 0.25-5.0 µg kg -1 for ten neonicotinoid insecticides and two metabolites. The average recoveries were in the range of 72.8-106.5%, with relative standard deviations (RSDs) below 13.8%, measured at three concentration levels. The developed method exhibits a high sensitivity and suitable for the simultaneous determination of neonicotinoid insecticides and metabolites in honey and royal-jelly., (Copyright © 2018. Published by Elsevier Ltd.)

Neonicotinoid insecticide use has been suggested as a cause of honey bee colony decline; however, detection rates and concentrations of neonicotinoid insecticide residues in field-collected honey bees have been low. We collected honey bee and beebread samples from apiaries in agricultural, developed, and undeveloped areas during 2 years in Virginia to assess whether landscape type or county pesticide use was predictive of honey bee colony exposure to neonicotinoid insecticides. Trace concentrations of the neonicotinoid imidacloprid were detected in honey bees (3 of 84 samples, 2.02-3.97 ng/g), whereas higher concentrations were detected in beebread (5 of 84 samples, 4.68-11.5 ng/g) and pollen (three of five pollen trap samples, 7.86-12.6 ng/g). Imidacloprid was only detected in samples collected during July and August and was not detected in honey bees from hives where neonicotinoids were detected in pollen or beebread. The number of hives sampled at a site, county pesticide use, and landscape characteristics were not predictive of neonicotinoid detections in honey bees or beebread (all P > 0.05). Field surveys may underestimate honey bee exposure to field-realistic levels of pesticides or the risk of exposure in different landscapes because of low detection rates. Undetectably low levels of exposure or high levels of exposure that go undetected raise questions with regard to potential threats to honey bees and other pollinators.