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Sperm fertility decreases significantly after freezing. Providing a suitable and useful diluent compound for freezing ram sperm can increase the efficiency of artificial insemination and consequently, the reproductive performance of sheep. Various biological properties such as antibacterial, anti-cancer, immunosuppressive, antioxidant and reproductive properties of royal jelly (RJ) are well known. The aim of this study was to investigate the possible synergistic effect of royal jelly in combination with glycerol and dimethyl sulfoxide (DMSO) in sperm cryopreservation extender of Romanov ram. The pooled semen samples from 5 Romanov rams were allocated into 3 experiments. The effect of 6% DMSO, 6% glycerol and a combination of 3% DMSO +3% glycerol co-supplemented with 1, 2 and 3% RJ was evaluated in 3 experiments. Samples were frozen by conventional slow freezing method and post-thaw parameters of total motility, progressive motility, plasma membrane integrity, DNA damage, apoptosis, enzyme activity of superoxide dismutase (SOD) and glutathione peroxidase (GPx), total antioxidant capacity (TAC) and malondialdehyde (MDA) were evaluated. The results showed that the percentage of motility, progressive motility, TAC, GPx, SOD and all sperm kinematic parameters except LIN in the group containing 2% RJ + 6% DMSO was higher than the control group (p 0.05). Some parameters such as progressive motility, sperm membrane integrity, TAC, GPx, VAP, VCL, STR and SRT in the group containing 2% RJ + 6% DMSO were more than (p 0.05) in the sperm group containing 1% RJ + 6% DMSO. MDA values in sperm groups containing 2% RJ + 6% DMSO were significantly (p 0.05) lower than the sperm containing 1% royal jelly and the control group. In the sperm group containing 2% RJ + 6% glycerol, sperm membrane integrity, TAC, GPx, SOD, progressive motility and all sperm kinematic parameters except VAP were higher and MDA values and sperm abnormalities were lower than the control group (p 0.05). The sperm group containing 1% RJ and 3% DMSO +3% glycerol had higher motility, progressive motility, membrane integrity, and all sperm kinematic parameters except VSL; and lower sperm abnormalities, DNA damage, apoptosis and MDA than the control group (p 0.05). As a general conclusion of this study, the addition of 2% RJ + 3% DMSO and 3% glycerol to the freezing extender improved microscopic and biochemical ram sperm parameters after the freeze-thaw process. Hence, moderate concentrations of royal jelly (2%) are sufficient to protect sperm from freezing damage, and high (3%) and low (1%) concentrations do not have a good cryoprotective effect.

Objective Genitourinary syndrome of menopause is a progressive condition characterized by a decrease in estrogen, which causes bothersome genital symptoms. The purpose of the study was to determine the effect of royal jelly on the genitourinary syndrome of menopause. Materials and methods A randomized controlled trial was carried out from November 2018 to June 2019 in Bandar Abbas, Iran. The trial was registered in the Iranian Registry of Clinical Trials (1RCT20181107041585N1) with the main objective of determining if royal jelly could reduce the genitourinary syndrome of menopausal women. Eligible women were randomly assigned to receive either daily 1g of oral royal jelly or placebo for 8 weeks. The urogenital subscale of the Menopausal Rating Scale was used to determine genitourinary syndrome. Independent samples t-test was used for inter-group comparisons and paired samples t-test for pre-and post-treatment comparisons. Results There were no differences in the severity of sexual problems, bladder complications, or vaginal dryness between groups before intervention. Although the intervention group's bladder complications improved slightly after eight weeks of royal jelly treatment compared to the control group (p ​= ​0.04), there were no significant changes in vaginal dryness, sexual problems, or total urogenital score. The within-group changes (before and after treatment) also showed no differences in urogenital symptoms. Conclusions A daily dose of 1g royal jelly taken orally for 8 weeks did not alleviate menopausal genitourinary syndrome. No serious side effects were observed. To make more reliable decisions about the use and safety of royal jelly in the future, different doses of royal jelly and longer trials are required.

The dominant protein components of honeybee royal jelly (RJ) are major royal jelly proteins (MRJPs), which exhibit various biological properties. However, the biological basis of why bee venom contains MRJPs and what role MRJPs play in bee venom remains to be elucidated. This study reports the antiapoptotic role of MRJP 8 of Apis mellifera venom (AmMRJP 8) in melittin-treated mammalian cells. Recombinant AmMRJP 8 reduced caspase-3 activity and melittin-induced cell apoptosis. Additionally, recombinant AmMRJP 8 decreased the production levels of H2O2 and proinflammatory molecules. These results indicate that MRJP in bee venom plays a role in cell protection in bee venom-induced inflammatory responses.

Honey bee is vital for pollination and ecological services, boosting crops productivity in terms of quality and quantity and production of colony products: wax, royal jelly, bee venom, honey, pollen and propolis. Honey bees are most important plant pollinators and almost one third of diet depends on bee’s pollination, worth billions of dollars. Hence the role that honey bees have in environment and their economic importance in food production, their health is of dominant significance. Honey bees can be infected by various pathogens like: viruses, bacteria, fungi, or infested by parasitic mites. At least more than 20 viruses have been identified to infect honey bees worldwide, generally from Dicistroviridae as well as Iflaviridae families, like ABPV (Acute Bee Paralysis Virus), BQCV (Black Queen Cell Virus), KBV (Kashmir Bee Virus), SBV (Sacbrood Virus), CBPV (Chronic bee paralysis virus), SBPV (Slow Bee Paralysis Virus) along with IAPV (Israeli acute paralysis virus), and DWV (Deformed Wing Virus) are prominent and cause infections harmful for honey bee colonies health. This issue about honey bee viruses demonstrates remarkably how diverse this field is, and considerable work has to be done to get a comprehensive interpretation of the bee virology.

Graphical abstract, Introduction In recent years, analytical screening methods for simultaneous detection of multivitamins have gained substantial attention to ensure quality and public confidence in dietary supplements. Even so, few analytical methods have been proposed for simultaneous analysis of multivitamin constituents due to the large divergence in chemical characteristics. Objectives In the present study, the objective was to develop a simple and rapid direct nano-electrospray ionization-tandem mass spectrometry (DI-nano-ESI-MS/MS) method for targeted detection of water soluble vitamins, fat soluble vitamins, amino acids, royal jelly, ginkgo biloba, and ginseng in a dietary supplement. The applicability of dilute-and-shoot-based DI-nano-ESI-MS/MS to analyze the same tested compounds and their related metabolites in clinical samples was also examined. Methods Intact urine mixed with the ionization solvent was loaded (4-μL aliquot) into a nanospray (NS) capillary of 1-μm tip diameter. The NS capillary was then fitted into an off-line ion source at a distance of 5 mm from MS aperture. The sample was directly injected by applying a voltage of 1.1 kV, producing a numerous of m/z peaks for analysis in mere minutes. Results The DI-nano-ESI-MS/MS method successfully identified almost all dietary supplement components, as well as a plethora of component-related metabolites in clinical samples. In addition, a new merit of the proposed method for the detection of index marker and chemical contaminants as well as subspecies identification was investigated for further quality evaluation of the dietary supplement. Conclusions The previous findings illustrated that DI-nano-ESI-MS/MS approach can emerge as a powerful, high throughput, and promising analytical tool for screening and accurate detection of various pharmaceuticals and ingredient in dietary supplements as well as biological fluids.

The neuronal basis of complex social behavior is still poorly understood. In honeybees, reproductive investment decisions are made at the colony-level. Queens develop from female-destined larvae that receive alloparental care from nurse bees in the form of ad-libitum royal jelly (RJ) secretions. Typically, the number of raised new queens is limited but genetic breeding of "royal jelly bees" (RJBs) for enhanced RJ production over decades has led to a dramatic increase of reproductive investment in queens. Here, we compare RJBs to unselected Italian bees (ITBs) to investigate how their cognitive processing of larval signals in the mushroom bodies (MBs) and antennal lobes (ALs) may contribute to their behavioral differences. A cross-fostering experiment confirms that the RJB syndrome is mainly due to a shift in nurse bee alloparental care behavior. Using olfactory conditioning of the proboscis extension reflex, we show that the RJB nurses spontaneously respond more often to larval odors compared with ITB nurses but their subsequent learning occurs at similar rates. These phenotypic findings are corroborated by our demonstration that the proteome of the brain, particularly of the ALs differs between RJBs and ITBs. Notably, in the ALs of RJB newly emerged bees and nurses compared with ITBs, processes of energy and nutrient metabolism, signal transduction are up-regulated, priming the ALs for receiving and processing the brood signals from the antennae. Moreover, highly abundant major royal jelly proteins and hexamerins in RJBs compared with ITBs during early life when the nervous system still develops suggest crucial new neurobiological roles for these well-characterized proteins. Altogether, our findings reveal that RJBs have evolved a strong olfactory response to larvae, enabled by numerous neurophysiological adaptations that increase the nurse bees' alloparental care behavior.

Background According to the World Health Organization, two billion people will attain the age of 60 years or more by 2050. Ageing is a major risk factor for a number of neurodegenerative disorders, which currently possess challenge to the global health status, carrying economic and social consequences. Therefore, attention has been dedicated towards the development of neuroprotective agents derived from natural sources. Honeybee products, such as honey, bee pollen, bee bread, propolis, royal jelly, beeswax, and bee venom have been used for therapeutic purposes since ancient times in Egypt, Greece, and China. Despite the emergence of modern medicine, bee products remain clinically relevant owing to their potential as anti-inflammatory, anti-oxidant, and neuroprotective agents. Scope and approach This review demonstrates the potential of bee products against neurological disorders in the light of the current literature. Key findings and conclusions Bee products and individual isolated components have enormous therapeutic potential for multiple neurological disorders. The different studies show overall neuroprotective and nerve-tonic characteristics of bee products, mainly due to their anti-oxidant, anti-inflammatory and anti-apoptotic features. However, some limitations such as allergic reactions and the cytotoxic effect of some bee products warrant a special care in its development as drug leads in future studies.

Major royal jelly proteins (MRJPs) of honeybee royal jelly (RJ) exhibit antimicrobial and antioxidant activities. Although MRJPs of Apis mellifera RJ (AmMRJPs) responsible for antibacterial activity have been identified, AmMRJPs with antioxidant effects remain to be elucidated. Here we identified and compared the antioxidant activities of purified recombinant AmMRJPs 1–7, which are expressed in baculovirus-infected insect cells. Antioxidant assays of recombinant AmMRJPs 1–7 against H2O2 revealed that AmMRJPs reduce caspase-3 activity and oxidative stress-induced cell apoptosis and lead to increased cell viability. Consistent with these results, AmMRJPs 1–7 exhibit 1,1-diphenyl-2-picrylhydrazyl radical-scavenging activity and protect against oxidative DNA damage. These results indicate that AmMRJPs play a role as antioxidants in A. mellifera RJ.

Royal jelly (RJ) is a well-known functional and medicinal food for human health promotion. Major royal jelly proteins (MRJPs), which are the major protein components in RJ, exhibit antimicrobial activities. However, the identities of the MRJPs of RJ responsible for its antioxidant effects have remained unclear. Here, we report that honeybee (Apis cerana) MRJP 2 (AcMRJP2) acts as an antimicrobial and antioxidant agent in RJ. Using recombinant AcMRJP2, which was produced in baculovirus-infected insect cells, we established the antimicrobial and antioxidant roles of MRJP 2. AcMRJP2 bound to the surfaces of bacteria, fungi, and yeast, which then induced structural damage in the microbial cell walls and led to a broad spectrum of antimicrobial activities. AcMRJP2 protected mammalian and insect cells via the direct shielding of the cell against oxidative stress, which led to reduced levels of caspase-3 activity and oxidative stress-induced cell apoptosis, followed by increased cell viability. Moreover, AcMRJP2 exhibited DNA protection activity against reactive oxygen species (ROS). Our data indicate that AcMRJP2 could play a crucial role as an antimicrobial agent and antioxidant in RJ, suggesting that MRJP 2 is a component responsible for the antimicrobial and antioxidant activities of RJ.

We reared the cross-bred FD lineage (F♀ × D♂) from inbred lines in pursuit of high royal jelly (RJ)-producing honeybee traits. The present study compared the new crossbred FD line to a high RJ-producing colony (HRC) and a low RJ-producing colony (LRC) by investigating the production of RJ, morphological characteristics, and age-dependent hypopharyngeal gland (HPG) development of the honeybees. The HRC exhibited the highest RJ production (416.46 ± 71.49 mg/cup), followed by the FD (369.29 ± 80.89 mg/cup), and the LRC (284.79 ± 38.54 mg/cup) colonies, respectively. The 10-HDA content was highest in the FD colony (2.38%), followed by the LRC (2.06%) and the HRC (1.91%). Compared to newly emerged bees, the body weight, fresh head weight, and HPG development increased until 9 days, and then decreased thereafter. Moreover, the length of the acini of the HPG at 9 days-old was almost twice as long than those of the bees at first emergence in all colonies. The acini diameters of the HPG in the HRC were significantly larger at all age stages. Compared to the LRC line, the acini in the FD line were significantly longer until 9 days after hatching. The abdominal length and forewing length were longer in the HRC, while the corbicular area, and head, proboscis, and glossa lengths were longer in the FD. The discriminant function analysis revealed that 78.1% of the individuals were classified correctly. Further assessements of wing geometric morphometrics are needed to clarify the morphological characteristics of the new hybrid FD line with high RJ-producing traits.

Major royal jelly proteins (MRJPs) are the protein components in royal jelly (RJ). MRJPs 1–7 are detected in the honeybee Apis mellifera RJ. Although A. mellifera MRJP (AmMRJP) 2 exhibited antibacterial activity, the other MRJPs with antimicrobial activities in A. mellifera RJ remains largely unknown. Here, we compared the antibacterial activity of recombinant AmMRJPs 1–7 expressed in baculovirus-infected insect cells. Antibacterial assays of recombinant AmMRJPs 1–7 against the gram-negative bacterium Escherichia coli revealed that AmMRJPs 2–5 and 7 exhibited antibacterial activity, whereas AmMRJPs 1 and 6 displayed almost no antibacterial activity. Consistent with the antibacterial activity of AmMRJPs, AmMRJPs 2–5 and 7 are bound to bacterial cell walls. These results indicated that AmMRJPs 2–5 and 7 contribute directly to the antibacterial property of RJ, suggesting that MRJPs play a role in the antimicrobial property of RJ.

The protein component of honeybee royal jelly (RJ) is constituted by major royal jelly proteins (MRJPs). The Asiatic honeybee (Apis cerana) MRJP-4 (AcMRJP4) exhibits antimicrobial activities. In this study, we identified the antimicrobial activity of AcMRJP4-15, which is a hydrophilic peptide with 88 amino acid residues in the C-terminal of AcMRJP4 that contains a high content of Asn and positively charged amino acids. Recombinant AcMRJP4-15, which is expressed as a 15-kDa peptide in baculovirus-infected insect cells, induced structural damage to the cell walls of bacteria, fungi, and yeast. Interestingly, the antimicrobial activity of AcMRJP4-15 was greater than that of AcMRJP4, demonstrating that the antimicrobial activity of AcMRJP4 was due in large part to the C-terminal. Our data suggest that AcMRJP4-15 can function as an effective antimicrobial agent.

Background and purpose: Apitherapists promote the medical use of products from the beehive (bee venom, propolis, pollen, honey, royal jelly, dead bees, apilarnil, wax, wax moths), the use of beehive air or therapeutic sleep on a beehive. However, little is known about how far such treatment options are perceived as acceptable by patients. Methods Patients visiting either a family doctor in Kehl or a gynecologist in Weilburg (both in Germany) were asked to rate their knowledge of apitherapy as well as their readiness to use apitherapeutic measures. Results Honey and propolis represent the best-known bee products whereas beehive air and apilarnil are greatly unknown to the patients. Only honey seems to be an acceptable treatment option whereas propolis, pollen and royal jelly seem to be less acceptable. Bee venom was not considered an interesting treatment possibility and, in particular, live bee stings were considered less desirable. This study found that gender and acquaintance with a beekeeper influenced the patients’ ratings but age, education and current medical condition did not. Conclusion Live bee stings, apilarnil or the inhalation of beehive air are not appealing to the majority of patients. Before apitherapeutic methods are promoted, it seems to be important to know about patients’ willingness to tolerate such treatments. Perhaps therapy modifications can be offered which seem more acceptable.

An ion chromatography coupled with pulsed amperometric detection (IC-PAD) method was developed for the determination of sugars in royal jelly (RJ). The method enables quantification of 19 sugars with higher sensitivity than HPLC method. Good linearity was observed in the range of 0.02–500 mg L–1 with coefficients of determination (R2) ≥ 0.9994. The limits of detection (LODs) and limits of quantification (LOQs) were in the range of 1.0–110 μg L–1 and 3.7–380 μg L–1, respectively. The average recoveries ranged from 91.0% to 108.0%. This simple method can be an effective alternative to GC method, which requires complicated purification and derivatization steps. It was used for the analysis of 100 Chinese RJ samples from three different areas and the results of analyzed sugars were found to be in agreement with those reported in literatures. The sugar profile results have suggested that some minor sugars are better markers for discrimination in relation to geographical origin.

Liver diseases are serious life-threating conditions that should be controlled. Here, we identify a protein fraction from royal-jelly (RJ) that represents the most effective composite against CCl4-induced hepatotoxicity and HepG2 cell growth. Two closely related proteins were purified from this fraction by a new simple method and identified by MALDI-TOF MS as major RJ protein 2 (MRJP2) and its predicted isoform X1. The in silico assessment (3D structures and functions) of these proteins were performed using Iterative Threading ASSEmbly Refinement (I-TASSER) analysis and RAMPAGE program. These two purified proteins were able to relieve the necrotic hepatocytes (by 60.4%) via reducing tumor necrosis factor (TNF)-α, mixed lineage kinase domain-like protein (MLKL) and intracellular reactive species. The latter effects associated with improving hepatocyte functions. Furthermore, they revealed the potent anticancer effect via induction of caspase-dependent apoptosis and controlling the expression of both Bcl-2 and p53 in HepG2 cells. Thus, MRJP2 and its isoform X1 can be a promising dual strategy for fighting hepatic injury and cancer in future animal and human studies.

Introduction For hundreds of years honey and bee products have been used in diets and as a home remedies. In order to assess the prevalence of bee products usage in pediatric pulmonary patients, an anonymous survey of parents/guardians was conducted. Methods A cross-sectional study was performed in the pediatric pulmonology outpatients clinic using a questionnaire which contained 20 open-ended and closed questions. Results Of 138 questionnaires distributed, 120 were fully completed and included in the analysis. 79% of the respondents had given their child some form of bee product to alleviate health problems (e.g. asthma and bronchitis), which most frequently included meadow honey, propolis and royal jelly. Living in the city and older age of parents/guardians significantly increased the probability of using bee products. Products were usually purchased in pharmacies or beekeepers shops. Bee product blends, used by ≈74%, were often produced at home, and they contained a variety of plant ingredients (pine needles, black radish, dandelions, onions, ginger, thyme, St Johns’ Wort, yarrow, etc.). More than a half reported better improvements combining bee products and prescribed medicines. A few very mild side-effects were reported by 5% and one case of an allergic reaction to propolis. Conclusions Medicinal usage of bee products among pediatric pulmonology patients in South- East Serbia is widespread. Almost all interviewed parents/guardians would like to learn more about bee products and the possibilities for their safe and effective application.

Major royal jelly proteins (MRJPs) are important protein components of bee royal jelly (RJ) and exhibit various biological and pharmacological activities. The antimicrobial activities of the royalisin and the jelleines contained within MRJP 1 and MRJP 2 in RJ have been elucidated. However, the antimicrobial effects of other MRJPs remain largely unknown. In this study, we demonstrated the antimicrobial activity of the Asiatic honeybee (Apis cerana) MRJP 4 (AcMRJP4). Recombinant AcMRJP4 was expressed as a 63-kDa protein in baculovirus-infected insect cells. We examined the antimicrobial activity of recombinant AcMRJP4 against bacteria, fungi, and yeast. The mechanisms underlying the antimicrobial activity of AcMRJP4 were assessed using western blot analysis, immunofluorescence staining, and scanning electron microscopy. Recombinant AcMRJP4 bound to the cell walls of bacteria, fungi, and yeast and induced structural damage in the microbial cell walls. AcMRJP4 has an antimicrobial role and exhibits a broad spectrum of antimicrobial activities against bacteria, fungi, and yeast. We demonstrated that AcMRJP4 functions as an antimicrobial agent with activity against bacteria, fungi, and yeast. Together, our data identified a novel function of MRJP 4 as an antimicrobial agent.

Female insects that survive a pathogen attack can produce more pathogen-resistant offspring in a process called trans-generational immune priming. In the honey bee (Apis mellifera), the egg-yolk precursor protein Vitellogenin transports fragments of pathogen cells into the egg, thereby setting the stage for a recruitment of immunological defenses prior to hatching. Honey bees live in complex societies where reproduction and communal tasks are divided between a queen and her sterile female workers. Worker bees metabolize Vitellogenin to synthesize royal jelly, a protein-rich glandular secretion fed to the queen and young larvae. We ask if workers can participate in trans-generational immune priming by transferring pathogen fragments to the queen or larvae via royal jelly. As a first step toward answering this question, we tested whether worker-ingested bacterial fragments can be transported to jelly-producing glands, and what role Vitellogenin plays in this transport. To do this, we fed fluorescently labelled Escherichia coli to workers with experimentally manipulated levels of Vitellogenin. We found that bacterial fragments were transported to the glands of control workers, while they were not detected at the glands of workers subjected to RNA interference-mediated Vitellogenin gene knockdown, suggesting that Vitellogenin plays a role in this transport. Our results provide initial evidence that trans-generational immune priming may operate at a colony-wide level in honey bees.

Thanks to the bioactive components it contains, royal jelly has many positive effects on health as well as being nutritious. In the study, royal jelly at a rate of 2% (w/v) to raw cow milk whose dry matter was adjusted to 13%, and S thermophilus, Lb. bulgaricus and Bifidobacterium bifidum added probiotic yoghurt (PYA) with increased functional properties was produced. In the other part, probiotic yoghurt (PY) was produced without adding royal jelly. Physicochemical, microbiological, rheological, sensory tests and color analysis were performed on the samples, and the fatty acid composition and the amount of trans-10-hydroxy-2-decenoic acid (10-HDA) were determined in the yoghurt samples. In the study, the association between the increase in acidity, decrease in carbohydrate level, dry matter level, rheological properties, the viability of probiotics, fatty acid composition, and sensory properties during storage and royal jelly addition was found to be significant (p < 0.05). It was determined that PYA samples were more acceptable than PY samples in terms of important parameters such as rheological properties, the viability of probiotic cultures, and fatty acid composition. It was determined that the amount of trans-10-hydroxy-2-decenoic acid (10-HDA), one of the most important bioactive components of royal jelly, was above the minimum level specified in the standards in yoghurt samples. Probiotic yoghurts produced with the addition of royal jelly can be a good alternative to innovative functional foods with acceptable product properties.

Royal jelly (RJ) is a highly valued natural product which has long been used as a functional food in various parts of the world. The aim of this study was to evaluate the effects of RJ supplementation (0.5%, 1%, and 1.5%, w/w) on physicochemical properties (i.e., viscosity, chemical composition, water holding capacity, antioxidant activity), organoleptic characteristics, culture viability, and biological activities (i.e., anticancer and antibacterial activities) of fermented milk. Results showed RJ supplementation could reduce the fermentation time, increase the viscosity and water holding capacity, and also enhance the antioxidant acitivity of fermented milk. Compared to the fermented milk without RJ addition, fermented milk with 1% RJ had significantly more probiotics Lactobacillus helveticus Lh-B02, approximately 9.73 log CFU/g during 21 days of cold storage. Additionally, RJ supplementation could greatly boost the bioactivities of fermented milk. Fermented milk with 1.5% RJ had the highest anticancer and antibacterial properties, whereas the one with 1% RJ had the best sensory attribute score. Overall, RJ has a great potential to be a viable dietary additive for the development of innovative dairy products with multifacet nutritional and bioactive properties.

Royal jelly has received attention because of its necessity for the development of queen honeybees as well as claims of benefits on human health; this product of the hypopharyngeal glands of worker bees contains a large number of proteins, some of which have been claimed to have various biological effects only in their glycosylated state. However, although there have been glycomic and glycoproteomic analyses in the past, none of the glycan structures previously defined would appear to have potential to trigger specific biological functions. In the current study, whole royal jelly as well as single protein bands were subject to off-line LC-MALDI-TOF MS glycomic analyses, complemented by permethylation, Western blotting and arraying data. Similarly to recent in-depth studies on other insect species, previously overlooked glucuronic acid termini, sulfation of mannose residues and core β-mannosylation of the N-glycans were found; additionally, a relatively rare zwitterionic modification with phosphoethanolamine is present, in contrast to the phosphorylcholine occurring in lepidopteran species. Indicative of tissue-specific remodelling of glycans in the Golgi apparatus of hypopharyngeal gland cells, only a low amount of fucosylated or paucimannosidic glycans were detected as compared with other insect samples or even bee venom. The unusual modifications of hybrid and multiantennary structures defined here may not only have a physiological role in honeybee development, but represent epitopes recognized by pentraxins with roles in animal innate immunity.

In this study, new methods have been proposed to determine seven neonicotinoid insecticides (dinotefuran, nitenpyram, thiamethoxam, clothianidin, imidacloprid, acetamiprid and thiacloprid) in royal jelly-based products, by means of ultra-high-performance liquid chromatography coupled to a quadrupole-time-of-flight mass detector. Efficient sample treatments (with average analyte recoveries between 83% and 109%) involving solid-phase extraction (polymeric sorbent, Strata® X) and a dispersive liquid–liquid microextraction, were proposed to determine these neonicotinoids in liquid dietary supplements containing freeze-dried royal jelly and fresh royal jelly, respectively. Chromatographic analysis (8 min) was performed on a core-shell technology-based column (Kinetex® EVO C18). Both methods were fully validated and the data demonstrated that they are consistent, reliable and have a wide linear range of applicability. Low limits of quantification, ranging from 2.5 to 9.5 μg/kg, were obtained in all cases, and it was necessary to employ matrix-matched standards for correct quantification of three of the compounds in each of the royal jelly-based products. Finally, the proposed methods were applied to neonicotinoid analysis of royal jelly-based products from different Spanish regions.

Apis cerana cerana, an important endemic honey bee species in China, possesses valuable characteristics such as a sensitive olfactory system, good foraging ability, and strong resistance to parasitic mites. Here, we performed transcriptome sequencing of the antenna, the major chemosensory organ of the bee, using an Illumina sequencer, to identify typical differentially expressed genes (DEGs) in adult worker bees of different ages, namely, T1 (1 day); T2 (10 days); T3 (15 days); and T4 (25 days). Surprisingly, the expression levels of DEGs changed significantly between the T1 period and the other three periods. All the DEGs were classified into 26 expression profiles by trend analysis. Selected trend clusters were analyzed, and valuable information on gene expression patterns was obtained. We found that the expression levels of genes encoding cuticle proteins declined after eclosion, while those of immunity-related genes increased. In addition, genes encoding venom proteins and major royal jelly proteins were enriched at the T2 stage; small heat shock proteins showed significantly higher expression at the T3 stage; and some metabolism-related genes were more highly expressed at the T4 stage. The DEGs identified in this study may serve as a valuable resource for the characterization of expression patterns of antennal genes in A. cerana cerana. Furthermore, this study provides insights into the relationship between labor division in social bees and gene function.

Royal jelly (RJ) is known as a functional food for its diverse health-beneficial properties and complicated chemical compositions. Trans-10-hydroxy-2-decenoic acid (10-HDA) is the exclusive lipid component in RJ. In the present study, the in vitro anti-inflammatory effect of 10-HDA in LTA (Lipoteichoic acid from Staphylococcus aureus) induced RAW 264.7 macrophages are evaluated. The results showed that 10-HDA had potent, dose-dependent inhibitory effects on the release of major inflammatory mediators and NO. Several key inflammatory genes, including IL-1β, IL-6, MCP-1 and COX-2 have also been suppressed by 10-HDA. Furthermore, the effects of 10-HDA on LTA-induced pulmonary damage were also examined in mice. It was found that the administration of 10-HDA (100 mg/kg) can provide protective effects by attenuating lung histopathological changes and modulating the secretion of LTA-stimulated inflammatory cytokines in mice, such as IL-10, MCP-1 and TNF-α. Conclusively, the results reveal the potent anti-inflammatory properties of 10-HDA and provide biological information for the future application.

Royal Jelly (RJ) is a nutrient rich substance secreted by the hypopharyngeal glands of young worker bees. Owing to the exceptional biological properties, RJ is used in pharmaceutical, food and cosmetics industries. Several studies vouch that RJ has anti-ageing, antibacterial, anti-fatigue, anti-inflammatory, antioxidant, antitumor, anti-diabetic and antimutagenic potentials. These activities are mainly attributed to bioactive components it contains. One of the principal bioactive components is Major Royal Jelly Proteins (MRJPs) which is considered to be a major factor in honey bee queen development. In this review, we explore large number of studies that have been undertaken to elucidate the functions and characterization of Royal Jelly proteins and peptides.

Royal Jelly (RJ) is a unique substance obtained from bees that has been used widely in European and Asian traditional medicine for its potential to prevent signs of aging through its antioxidative, anti-inflammatory, anti-hyperglycemic and anti-hypercholesterolemic properties. We recently reported an enhancement in spatial memory along with changes in monoaminergic transmission in aged rats after chronic RJ administration. Here, we aim to further explore the action of RJ on central nervous system activity by examining levels of amino acids in selected brain structures of aged male Wistar rats following 2-months of Greek RJ administration. RJ powder was previously chemically characterized and given orally (50 or 100 mg of powder/kg b.w./day) by gastric gavage. The concentrations of amino acids (alanine, aspartic acid, gamma-aminobutyric acid, glutamic acid, histidine and taurine) in the brain regions examined (prefrontal cortex, hippocampus, striatum and hypothalamus) were quantified using HPLC. We also examined basic biochemical parameters of renal and hepatic activity, as damage of these organs could potentially explain the changes in brain function and behavior. Upon biochemical examination, a decrease in the concentration of gamma-aminobutyric acid was observed in both the striatum and hypothalamus. Liver and kidney functions were not changed by chronic RJ-administration. Our results provide insight toward understanding the mechanism of action of RJ and its effects on neurotransmission in the central nervous system.

Summary Introduction Identification of thermogenic food ingredients is potentially a useful strategy for the prevention of obesity and related metabolic disorders. It has been reported that royal jelly (RJ) supplementation improves insulin sensitivity; however, its impacts on energy expenditure and adiposity remain elusive. We investigated anti-obesity effects of RJ supplementation and their relation to physical activity levels and thermogenic capacities of brown (BAT) and white adipose tissue (WAT). Methods C57BL/6J mice were fed under four different experimental conditions for 17 weeks: normal diet (ND), high fat diet (HFD), HFD with 5% RJ, and HFD with 5% honey bee larva powder (BL). Spontaneous locomotor activity, hepatic triglyceride (TG) content, and blood parameters were examined. Gene and protein expressions of thermogenic uncoupling protein 1 (UCP1) and mitochondrial cytochrome c oxidase subunit IV (COX-IV) in BAT and WAT were investigated by qPCR and Western blotting analysis, respectively. Results Dietary RJ, but not BL, suppressed HFD-induced accumulations of WAT and hepatic TG without modifying food intake. Consistently, RJ improved hyperglycemia and the homeostasis model assessment-insulin resistance (HOMA-IR). Although dietary RJ and BL unchanged locomotor activity, gene and protein expressions of UCP1 and COX-IV in BAT were increased in the RJ group compared to the other experimental groups. Neither the RJ nor BL treatment induced browning of WAT. Conclusion Our results indicate that dietary RJ ameliorates diet-induced obesity, hyperglycemia, and hepatic steatosis by promoting metabolic thermogenesis in BAT in mice. RJ may be a novel promising food ingredient to combat obesity and metabolic disorders.

In the present work a Hydrophilic Interaction Liquid Chromatography-tandem Mass Spectrometry (HILIC-MS/MS) method was developed for the efficient separation and quantification of a large number of small polar bioactive molecules in Royal Jelly. The method was validated and provided satisfactory detection sensitivity for 88 components. Quantification was proven to be precise for 64 components exhibiting good linearity, recoveries R% >90% for the majority of analytes and intra- and inter-day precision from 0.14 to 20% RSD. Analysis of 125 fresh royal jelly samples of Greek origin provided useful information on royal jelly's hydrophilic bioactive components revealing lysine, ribose, proline, melezitose and glutamic acid to be in high abundance. In addition the occurrence of 18 hydrophilic nutrients which have not been reported previously as royal jelly constituents is shown.

For sustainable and organic farming, organic manures such as cattle manure (CMn) and bio-nutritious substances such as royal jelly (RJ) should be used as complete or at least partial alternatives to chemical fertilizers (CFs). Therefore, two consecutive trial seasons (2018 and 2019) were conducted to study the effect of soil fertilization with CMn and/or CFs in combination with foliar spraying with RJ on growth, yield, and chemical composition of roselle (Hibiscus sabdariffa L.) plant and seed. The results indicated that spraying roselle plants with RJ resulted in a significant increase in all parameters studied over the control; however, this increase was enhanced when RJ was combined with soil fertilization. The highest values for most of the growth traits, chemical composition of seeds, sepal anthocyanin and vitamin C contents, and sepal acidity were obtained by applying 2 or 4 g RJ L−1 + (30 m3 CMn ha−1 + 50 % dose CFs). In addition, the highest values for yield components and plant chemical composition were obtained by applying 2 or 4 g RJ L−1 + (0 m3 CMn ha−1 + full dose CFs). The results also indicated that applying CMn in combination with RJ gave values mostly close to those of CFs treatment. In conclusion, the results recommend the use of RJ (2 g L−1 as a foliar feeding) along with CMn (30 m3 ha−1) as a partial alternative to chemical fertilizers for sustainably producing healthy roselle sepals.

© 2021 The Author(s)Royal jelly (RJ) is a natural product that is fed to bee queens during their entire life, and it contributes to their growth and development, fertility, and long lifespan. Although it is known that these unique effects of RJ, it has not been evaluated fed diet studies in fish until now. The study was carried out to determine the influence of RJ diets with different rates (0.0; 0.1; 0.4; 1.6 and 6.4 %) on expressions of growth (GH-I, IGF-I) and immune system (TGF-β) hormones of zebrafish (Danio rerio). Growth parameters was also studied and found to be higher in D4 group containing 1.6 % additives compared with other groups. RJ dietary supplementation showed significantly up-regulated in GH-1 gene expression in the liver tissues of fish fed D5 diet respect to the control group. Conversely, expression level was down-regulated in kidney tissues of group D4 compared with the control group (p < 0.05). IGF-1 gene expression noticeably increased in the muscle tissues of fish fed the D5 diet containing 6.4 % RJ compared to the control group. The greatest decrease was found in the muscle tissues of group D3 diet (p < 0.05). Although among the diet groups the highest amount of TGF-β expression was found in the muscle tissue of fish that were fed the D4 diet. It was seen that the mRNA level amounts obtained in all applications were below the level of the control group. These results indicate that the high-dose RJ included in the feed positively influenced the growth hormones of the zebrafish at transcriptomic level. To sum up, this study highlights the importance that 1.6 % or 6.4 % RJ included in the diet of zebrafish positively influenced their growth performance and growth hormone gene expression and may be considered a potential to be sustainable alternative for growth promoter in aquaculture practice.

Proteomics is an emerging tool in food authentication that has not been optimised for honey analysis. In this study, we present a qualitative proteomic analysis of New Zealand mānuka (Leptospermum scoparium) honey. A total of fifty bee-derived proteins were identified in the honey, the most predominant being major royal jelly proteins (MRJPs). We also demonstrate for the first time the presence of unique nectar-derived proteins in mānuka honey. A total of 17 mānuka plant proteins were identified, a-third of which were putative pathogenesis-related proteins. Two proteins involved in drought tolerance were also identified. Twelve candidate peptides were selected as potential authentication markers based on their uniqueness to mānuka honey. Nectar analyses confirmed the origin and specificity of these peptides to L. scoparium nectar, thus presenting peptide profiling as a viable and novel approach for mānuka honey authentication. Raw data are available via ProteomeXchange with identifier PXD021730.

Royal jelly (RJ) is attractive functional food due to health promoting effect and rich nutrient content. The aim of this study was to produce a probiotic dairy dessert (Keskul) at an optimal RJ dose which is no adverse effect on physicochemical, rheological, sensory properties and functionality of the product. Initially, the growth parameters of Lactobacillus plantarum, Lactobacillus rhamnosus and Bifidobacterium animalis subspp. lactis in presence of RJ (0.001–10.0 mg/mL) were evaluated with Gompertz model. Then, the experimental (100 mg/mL) and estimated (56 mg/mL) Minimum Inhibitory Concentration (MIC) of tested probiotic bacteria were determined according to predictive microbiology approach. Among probiotics, L. plantarum was found distinctively proper for producing dairy dessert with RJ. In this study, a new dairy dessert was developed consisting of RJ and L. plantarum. The control, including probiotic (D1), probiotic and RJ (1 mg/mL) (D2), probiotic and RJ (5 mg/mL) (D3) were analyzed during shelf life of dessert. The combination of L. plantarum with RJ (1 mg/mL) provided probiotic properties (>6 log CFU/g) and limited the mold count. Thus, the dairy dessert developed in this study may be a good alternative to innovative functional food using RJ with acceptable sensorial attributes.

Honey bee larvae exposed to sublethal doses of imidacloprid show behavioural abnormalities as adult insects. Previous studies have demonstrated that this phenomenon originates from abnormal neural development in response to imidacloprid exposure. Here, we further investigated the global gene expression changes in the heads of newly emerged adults and observed that 578 genes showed more than 2-fold changes in gene expression after imidacloprid exposure. This information might aid in understanding the effects of pesticides on the health of pollinators. For example, the genes encoding major royal jelly proteins (MRJPs), a group of multifunctional proteins with significant roles in the sustainable development of bee colonies, were strongly downregulated. These downregulation patterns were further confirmed through analyses using quantitative reverse transcription-polymerase chain reaction on the heads of 6-day-old nurse bees. To our knowledge, this study is the first to demonstrate that sublethal doses of imidacloprid affect mrjp expression and likely weaken bee colonies.

The aim of the present study was to evaluate different concentrations of royal jelly (RJ) supplemented extenders for post-thaw quality and incubation resilience of goat spermatozoa. Semen samples were collected from five goats. Pooled semen were diluted with soybean lecithin-based extender without RJ (control) or supplemented with different concentrations (0.25, 0.5 and 0.75%) of RJ (RJ0.25, RJ0.5, RJ0.75 respectively), at a final concentration of 150 x 106 spermatozoon/mL. Semen samples were assessed for sperm motility, plasma membrane integrity using hypoosmotic swelling test (HOST) damaged acrosome using FITC-Pisum sativum agglutinin (PSA-FITC) and DNA integrity using terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL). The addition of RJ (0.5%, 0.75%) led to higher percentages of subjective motilities (55.33 +/- 2.29%, 57.67 +/- 2.58%) compared to control and RJ0.25 groups (49.00 +/- 2,80%, 51.67 +/- 3.09%) (P < 0.05) following the freezethawing process. RJ0.5 and RJ0.75 groups had higher plasma membrane functional integrities (66.40 +/- 1.34%, 68.20 +/- 2.05%) and lower defected acrosome rates (24.60 +/- 3.36%, 23.80 +/- 2.27%) compared to the other groups (P < 0.05). DNA damaged spermatozoa in all groups were not significant (P > 0.05). In the end of incubation, motility and HOST rates of RJ0.5 (14.00 +/- 3.87%, 31.20 +/- 3.70%) and RJ0.75 (15.00 +/- 3.27%, 29.20 +/- 2.59%) groups were higher than control (8.00 +/- 2.54%, 18.20 +/- 3.11%) and RJ0.25 (9.00 +/- 2.07%, 20.60 +/- 2.88%) groups (P < 0.05). Also defected acrosome and DNA fragmation rates of RJ0.5 (32.20 +/- 1.30%, 5.4 +/- 0.55%) and RJ0.75 (29.20 +/- 1.30%, 5.80 +/- 0.45%) groups were significantly lower than control (38.80 +/- 0.84%, 7.40 +/- 1.34%) and RJ0.25 (39.80 +/- 2.05%, 7.00 +/- 1.58) groups. This study shows that RJ supplemented extenders have beneficial effect on goat sperm parameters at 0 h and 6 h of incubation.

Objective: To evaluate the clinical and histopathological effects of propolis and royal jelly compared to photobiomodulation therapies on an animal model of 5-fluorouracil (5-FU)-induced oral mucositis (OM). Study Design: In vivo, controlled and blinded experiments were performed. Seventy-two male Wistar rats were randomly allocated into 4 groups (n = 18): negative control (no treatment), photobiomodulation therapy, propolis gel, and royal jelly. On days 0 and 2, animals received an intraperitoneal injection of 5-FU. On days 3 and 4, the buccal mucosa was scratched. Therapy was initiated on day 5. Six animals were euthanized on days 8, 10, and 14. Clinical analysis was performed using photography, and histopathological evaluation was based on re-epithelization and inflammation scores. Results: On days 8 and 10, animals of photobiomodulation, propolis gel, and royal jelly groups showed improvement of OM compared to the control group (P

RNA flow between organisms has been documented within and among different kingdoms of life. Recently, we demonstrated horizontal RNA transfer between honeybees involving secretion and ingestion of worker and royal jellies. However, how the jelly facilitates transfer of RNA is still unknown. Here, we show that worker and royal jellies harbor robust RNA-binding activity. We report that a highly abundant jelly component, major royal jelly protein 3 (MRJP-3), acts as an extracellular non-sequence-specific RNA-aggregating factor. Multivalent RNA binding stimulates higher-order assembly of MRJP-3 into extracellular ribonucleoprotein granules that protect RNA from degradation and enhance RNA bioavailability. These findings reveal that honeybees have evolved a secreted dietary RNA-binding factor to concentrate, stabilize, and share RNA among individuals. Our work identifies high-order ribonucleoprotein assemblies with functions outside cells and organisms.

Peptides with a similar antioxidant and ACE-inhibitory activity of royal jelly (RJ) generated from Alcalase hydrolysated pollen (AHP) were predicted by Response Surface Methodology (RSM). The model equations were proposed according to the effects of time and enzyme concentration on the antioxidant and ACE-inhibitory activity. The optimum values for Alcalase concentration and hydrolysis time were 1.5% and 4 h, respectively. Later, AHP was prepared and deproteinised to be further analysed using size-exclusion chromatography (SEC). After SEC separation, fractions with the highest activity of ACE-inhibitory, DPPH radical scavenging and ferric-reducing power were purified by RP-HPLC. The highest ACE-inhibitory and DPPH scavenging activity of fractions was found 100% and 66.61%, respectively. The most active fractions were analysed by nano-liquid chromatography and mass spectrometry in tandem (nLC-MS/MS) and a total of 195 peptide sequences were identified. The origins of all peptides were herbal proteins and certain coincidences with previously described bioactive sequences were discussed., This work was funded by the Ministry of Science, Research and Technology of Iran and the Grant AGL2014-57367-R and FEDER funds from the Spanish Ministry of Economy, Industry and Competitiveness. Ramón y Cajal postdoctoral contract to LM is acknowledged. LC-MS/MS analysis was carried out by in the SCSIE University of Valencia Proteomics Unit (Spain), a member of ISCIII Proteo Red Proteomics Platform.

Major royal jelly proteins (MRJPs) have been used as endogenous biomarkers for honey authentication. However, no study on quantification of MRJPs in honey authentication was reported. In order to develop a new honey authenticity method based on quantitative analysis of MRJPs in honey, trypsin-digested peptides from MRJPs were examined by mass spectrometry (MS). Three peptides, YNGVPSSLNVISK, TLQMIAGMK, and LTVAGESFTVK were selected as specific markers for MRJP1, MRJP 2 and MRJP 3(MRJP1∼3), respectively. An ultra performance liquid chromatography-triple quadrupole MS (UPLC-TQMS) method used for quantification of MRJPs in honey was developed and validated. A database on the MRJP1∼3 contents of honey established by quantification of 70 authentic samples showed a potential application in honey authenticity identification. The analysis results of 10 commercial honey samples by UPLC-TQMS with reference to the MRJP1∼3 database of authentic honey were compared and confirmed by widely used isotope ratio mass spectrometry, a AOAC Official Method. In conclusion, the results showed our method, a proteomic-based UPLC-TQMS method, has a high accuracy, sensitivity and specificity for honey authenticity.

Pyrrolizidine alkaloids (PA) are secondary metabolites of plants, which are mostly found in the genus Senecio, Echium, Crotalaria, and Eupatorium. The presence of 1,2-unsaturated PA in foods is a concern to food regulators around the world because these compounds have been associated to acute and chronic toxicity, mainly in the liver. The intake foods with PA/PANO usually occur through accidental ingestion of plants and their derivatives, besides to products of vegetal-animal origin, such as honey. PA/PANO are transferred to honey by their presence in nectar, honeydew, and pollen, which are collected from the flora by bees. In addition to honey, other beekeeping products, such as pollen, royal jelly, propolis, and beeswax, are also vulnerable to PA contamination. In this context, this review provides information about chemical characteristics, regulation, and toxicity, as well as summarizes and critically discusses scientific publications that evaluated PA in honeys, pollens, royal jelly, and propolis.

The entomological source of honey is linked to its origin and may substantially contribute to the end products' price. For example, honey from native and/or non-domesticated species, such as Asian Apis dorsata and Apis cerana, is more valued and achieves higher prices compared to the honey from managed colonies of A. mellifera. Such types of honey are often subject to various fraudulent practices, such as mislabelling of the origin or admixing with lower-cost and low-quality honey which does not match the label declaration. The entomological origin may be determined through DNA-based techniques. Here, we report on new DNA-based tools for the correct identification of the DNA signatures of three economically important honeybee species: Apis mellifera, Apis cerana, and Apis dorsata. Initially, we constructed EPIC markers for a nuclear single-copy adenine nucleotide translocase (ANT) gene to explore the properties of interspecific variation in honeybee tissue samples. For confirmation of the samples’ identity, we used widespread mitochondrial tRNALeu – cox2 marker. Based on successful sequencing and phylogenetic analyses, we identified a short interspecifically variable part in the ANT gene for which we designed PCR primers that allow the successful amplification of fragmented DNA from sources like honey. Amplified fragments were differentially double digested by restriction endonucleases MnlI and HpaI, which resulted in specific restriction patterns observed on agarose gel electrophoresis. Verification of the approach was performed with 25 tissue samples and tested additionally with 20 honey samples and 1 royal jelly sample. Our results demonstrate the correct assignment of the samples to their entomological origin on the interspecific level. Furthermore, PCR amplification using the designed EPIC marker and double restriction protocol was demonstrated as useful in detecting mixed samples as well as two mislabelled honey samples claimed to belong to Asian honeybee Apis cerana. Additionally, EPIC markers are also useful in detecting heterozygosity, for example, in tracing paternal and maternal lines in contrast to abundantly used mitochondrial tRNALeu – cox2, which is inherited via maternal line only and may thus significantly contribute to the resolving of paternal relationships. We show that our method is not limited to honey but may also be applied to other honeybee products, such as royal jelly.

Honey traceability is an important topic, especially for honeydew honeys, due to the increased incidence of adulteration. This study aimed to establish specific markers to quantify proteins in honey. A proteomics strategy to identify marker peptides from bracatinga honeydew honey was therefore developed. The proteomics approach was based on initial untargeted identification of honey proteins and peptides by LC-ESI-Triple-TOF-MS/MS, which identified the major royal jelly proteins (MRJP) presence. Afterwards, the peptides were selected by the in silico digestion. The marker peptides were quantified by the developed targeted LC-QqQ-MS/MS method, which provided good linearity and specificity, besides recoveries between 92 and 100% to quantify peptides from bracatinga honeydew honey. The uniqueness and high response in mass spectrometry were backed by further complementary protein analysis (SDS-PAGE). The selected marker peptides EALPHVPIFDR (MRJP 1), ILGANVK (MRJP 2), TFVTIER (MRJP 3), QNIDVVAR (MRJP 4), FINNDYNFNEVNFR (MRJP 5) and LLQPYPDWSWTK (MRJP 7), quantified by LC-QqQ-MS/MS, highlighted that the content of QNIDVVAR from MRJP 4 could be used to differentiate bracatinga honeydew honey from floral honeys (p

Studying the sublethal effects of agrochemical pesticides on nontarget honeybees (Apis mellifera) is important for agricultural development. Carbendazim is a widely used broad-spectrum fungicide that inhibits mitotic microtubule formation and cell division. However, the impact of carbendazim on bee health and development has not been fully elucidated. Here, using proteomics approaches, we assessed in vitro the changes in the expression of functional proteins in the head of newly emerged adults following treatment with field concentration of carbendazim during the larval stage. Treatment with carbendazim severely altered 266 protein expression patterns in the heads of adults and 218 of them showed downregulation after carbendazim exposure. Notably, major royal jelly proteins, a crucial multifunctional protein family with irreplaceable function in sustaining the development of colonies, were significantly suppressed in carbendazim-treated bees. This result was verified in both head and hypopharyngeal gland of nurse bees. Moreover, visual and olfactory loss, immune functions, muscular activity, social behavior, neural and brain development, protein synthesis and modification, and metabolism-related proteins were likely inhibited by carbendazim treatment. Together, these results suggest that carbendazim is an environmental risk factor that likely weakens bee colonies, partially due to reduced expression of major royal jelly proteins, which may be potential causes of colony collapse disorder.

Royal jelly (RJ), a natural secretion of nurse bees, is a functional food with health-promoting properties. To investigate RJ quality changes with enhanced output and harvesting time, we compared metabolic profiles of RJ harvested at 24 h, 48 h, and 72 h after larval grafting into queen cells from Italian bees (ITBs) and high RJ-producing bees (RJBs) selected from ITBs. A total of 77 high-abundance compounds were identified with reverse-phase liquid chromatography– and hydrophilic interaction liquid chromatography–high-resolution mass spectrometry. Metabolite abundance, water content, and antibacterial activity were found largely unchanged between both bee strains, indicating that a similar RJ quality was achieved. Notably, the observed 10-hydroxy-2-decenoic acid content in RJBs (1.98%) and ITBs (2.41%) meet the standard for premium RJ in China. RJ harvested at 48 h and 72 h had similar metabolic profiles, indicating little influence of the two harvesting times on RJ quality. The harvesting at 24 h could lead to a higher RJ quality with higher abundance levels of many bioactive compounds and lower water content. Our data based mainly on RJ metabolic composition constitute evidence for mass production of high-quality RJ.

Royal Jelly (RJ), a honeybee hypopharyngeal gland secretion of young nurse and an exclusive nourishment for bee queen, has been used since ancient times for care and human health and it is still very important in traditional and folkloristic medicine, especially in Asia within the apitherapy. Recently, RJ and its protein and lipid components have been subjected to several investigations on their antimicrobial activity due to extensive traditional uses and for a future application in medicine. Antimicrobial activities of crude Royal Jelly, Royalisin, 10-hydroxy-2-decenoic acid, Jelleines, Major Royal Jelly Proteins against different bacteria have been reported. All these beehive products showed antimicrobial activities that lead their potential employment in several fields as natural additives. RJ and its derived compounds show a highest activity especially against Gram positive bacteria. The purpose of this Review is to summarize the results of antimicrobial studies of Royal Jelly following the timescale of the researches. From the first scientific applications to the isolation of the single components in order to better understand its application in the past years and propose an employment in future studies as a natural antimicrobial agent.

Pyriproxyfen (PPN) is an insect growth regulator (IGR) that interferes with insect metamorphosis. Although the side effects of PPN on honey bee larval/adult stages have been studied, the risk to honey bee larvae from PPN residue in the environment is still unclear. In this study, we evaluated the impact of PPN on larval honey bees in field colonies by using an in vivo feeding assay. Oral toxicity to adult honey bees were determined. Finally, influence on royal jelly production was also examined. For in vivo feeding assay, the highest observed PPN treatment caused 67% mortality during pupal stage and in the remaining bees, 62.3% showed abnormal eclosion. Reductions in hatching rate, capping rate and adult emergence rate and increased abnormal eclosion rate were found in the colonies fed with 10 ppm PPN syrup. Oral toxicity test revealed that adult honey bees were less susceptible to PPN. Moreover, PPN reduced not only queen cell acceptance rate but also yield of royal jelly in queen cells. These results indicate that PPN has negative impacts on both larval and adult honey bees and royal jelly production, especially under high PPN concentrations. Since PPN is harmful to the development of honey bee larvae and pupae in the natural environment, the issue of honey bee colony contamination by PPN should be addressed.

The immunosuppressive drug, utilized widely in Egypt, cyclosporine A was studied to evaluate its toxicity in male rats. Animals were divided into a control (untreated), 3 groups treated intraperitoneally with 20, 40 and 60 m/kg cyclosporine A for 5, 10 and 15 days, respectively and 3 groups treated intraperitoneally with 20, 40 and 60 mg/kg of cyclosporine A plus 100 mg/kg royal jelly administrated orally. Toxicity evaluation was carried out using two main endpoints: reproductive study (sperm morphology and count abnormalities) and biochemical changes in liver and testis (DNA amounts). The aim of this work is to study the protective role of royal jelly against sperm abnormalities in shape and count, and changes in DNA contents in liver and testis tissue induced in rats when treated by cyclosporine A with different doses (20–40–60 mg/kg/day) for 5, 10, and 15 days in male rats and how the royal jelly can repair this changes. Our results showed that sperm abnormalities induced by cyclosporine A included deviation from normal shape in head and tail. Abnormal heads contained amorphous head and banana-shaped head, whereas the abnormal tails included divided and coiled tails. It also induced an insignificant effect on the total sperm counts after 5 days of injection with the drug combined with royal jelly. DNA contents were determined in rat liver and testis cells to illustrate the mutagenic effect of cyclosporine A and how the royal jelly can modulate this effect. From these results we concluded that cyclosporine A toxicity was dose and time dependent and should be administrated under special precautions and medical supervision. Using of royal jelly in combination with cyclosporine A drug decreased its toxic effect, so it's considered as protector.

Methicillin Resistant Staphylococcus aureus (MRSA) has become increasingly a major problem and responsible for most of the hospital acquired infections in Egypt. Thus, there is an urgent need for agents with new therapeutic targets particularly those targeting bacterial virulence. MRSA isolates were investigated for biofilm formation, adhesion and invasion assays. Thereafter, the effect of garlic extract and royal jelly on those virulence factors was tested. These two agents were tested in vitro and in vivo using an animal model of MRSA skin infection. The garlic extract caused significant reduction in biofilm formation (60% median reduction) however, it had no significant effect on bacterial adhesion. Royal jelly reduced biofilm formation and bacterial adhesion significantly (≥50% median reduction). Animal study revealed that royal jelly could eradicate MRSA completely and promote wound healing even in the uninfected groups. However, the wound in case of garlic treated groups was dry and no exudates or pus was observed indicating its antiseptic effect. Conclusion The results implied the significant role of royal jelly not only in eradication of MRSA infection, but also in promoting the healing process in the wounds.