Royal jelly (RJ) is a popular functional food with a wealth of health-promoting effects. Over 90% of the global RJ is produced in China mainly by a high RJ-producing honeybee (RJB) strain that can accept and feed a great number of queen larvae for RJ production. To elucidate RJ changes due to queen cell numbers (QCNs), we compared the yield, larval acceptance rate, metabolic and proteomic profiles, and antioxidant activities of RJ from 1 to 5 strips of queen cells (64 per strip) in RJB colonies. As QCNs increased, the larval acceptance rate was not found to vary (p = 0.269) whereas the RJ weight per cell began to significantly decline in the 5-strip colonies (p < 0.05). Increased QCNs had a profound impact on RJ metabolic profiles and mainly reduced fatty acid levels. Remarkably, the 10-hydroxy-2-decenoic acid (10-HDA) content, a most important indicator of RJ quality, declined gradually from 2.01% in the 1-strip colonies to 1.52% in the 5-strip colonies (p < 0.001). RJ proteomic profiles were minimally altered and antioxidant activities were not significantly changed by QCNs. Collectively, the metabolomics and proteomics data and the antioxidant activity test represent a global evaluation of the quality of RJ produced with different QCNs. Our findings gain new insights into higher-quality RJ production using the high-yielding RJBs.

Royal jelly (RJ), a natural honeybee product, has a wide range of antibacterial activities. N-glycosylated major royal jelly protein 2 (N-MRJP2), purified from RJ, can inhibit the growth of Paenibacillus larvae (P. larvae, Gram-positive), a contagious etiological agent of the American foulbrood disease of honeybees. However, the inhibitory mechanism is largely unknown. Antibacterial assay and membrane proteome were conducted to investigate the inhibition capacity of RJ from different instar larvae and P. larvae treated by N-MRJP2, respectively. The similar antibacterial efficiency of RJ from different larval instar indicates that RJ is vital for the adaptive immune defense of small larvae. The killing of P. larvae by N-MRJP2 is achieved by disturbing the cell wall biosynthesis, increasing the permeability of cell membrane, hindering aerobic respiration, restraining cell division and inducing cell death. This demonstrates that RJ is critical for the passive immunity of immature larvae and N-MRJP2 can be used as natural antibiotic substance to resist P. larvae, even for other gram-positive bacteria. This constitutes solid evidence that RJ and N-MRJP2 have potentials as novel antibacterial agents.

Camellia bee pollen (CBP) is a major kind of bee product which is collected by honeybees from tea tree (Camellia sinensis L.) flowers and agglutinated into pellets via oral secretion. Due to its special healthcare value, the authenticity of its botanical origin is of great interest. This study aimed at distinguishing CBP from other bee pollen, including rose, apricot, lotus, rape, and wuweizi bee pollen, based on a non-targeted metabolomics approach using ultra-high performance liquid chromatography–mass spectrometry. Among the bee pollen groups, 54 differential compounds were identified, including flavonol glycosides and flavone glycosides, catechins, amino acids, and organic acids. A clear separation between CBP and all other samples was observed in the score plots of the principal component analysis, indicating distinctive metabolic profiles of CBP. Notably, L-theanine (864.83–2204.26 mg/kg) and epicatechin gallate (94.08–401.82 mg/kg) were identified exclusively in all CBP and were proposed as marker compounds of CBP. Our study unravels the distinctive metabolic profiles of CBP and provides specific and quantified metabolite indicators for the assessment of authentic CBP.