LC-MS/MS Unlocks 19th-Century Honeybee Queen Secrets

A team of researchers made up of members of the University of Copenhagen (Denmark), the Technical University of Denmark, and the University of Cambridge (United Kingdom) used liquid-chromatography-tandem mass spectrometry (LC-MS/MS) to examine a honeybee queen cell specimen from the 19th century. In their efforts to explore a potentially untapped resource within natural history collections—specimens related to historical beeswax—the researchers uncovered valuable insights into past bee-rearing practices. Their work may also help inform solutions to the challenges bees face today. The study was published in Open Research Europe (1).

The western honeybee, Apis mellifera, an insect known for its advanced level of social organization, is critical for its role in ecology, agriculture and economy (2,3). Although each bee has an important role in the colony, the research team believed that it may be valuable to focus their work specifically on queen bees specifically. The queen is the only bee without which the entire hive will not survive; she is the mother of all bees in the colony. Through the queen’s pheromones, the colony maintains order and functions in a manner that reflects the requirements of the colony. Human influence on queen rearing (such as raising the queen and replacing them within the hive) has been a prominent practice in beekeeping ever since the 19^th century, as the quality of the queen significantly affects the whole hive, including its reproductivity and resistance to disease (2,4).

Natural history museum collections, with preserved specimens and historical data, can provide critical insights into the evolution, health, and behaviors of honeybee populations over time. Through the study of past collections of Apis mellifera, researchers can identify the genetic diversity, disease patterns, and environmental impacts of the species, assisting in the development of effective conservation global strategies that can aid in their protection and continuance (5,6).

In this study, the intact and closed cell was analyzed by X-ray computed tomography (CT), and a perfectly preserved queen bee inside her cell was revealed to be present. Subsequently, a micro-destructive approach was used to evaluate the possibility of protein extraction for a palaeoproteomic (a term used to describe the applications of protein analysis to ancient materials) to ancient materials [7]). approach. Using LC-MS/MS, the team were able to recover 120 non-contaminant proteins, mostly bee-related, including major royal jelly proteins (MRJPs) crucial for queen nutrition and development, and silk fibroin proteins. Analysis of the protein modifications indicate their endogenous source and show the presence of in vivo glycosylation, which probably helped protein preservation and allows for the study of antimicrobial and metabolic functions of bees in the past (1).

The results of this study could indicate that the study of specimens such as the honeybee can provide biomolecular information and can influence future research on queen development, diet, and hive conditions. This data can present an important background for discovering how honeybee populations have adapted to changing environments and pressures over time. In addition, the findings can further highlight the scientific potential of natural history collections in the addressing of current questions concerning honeybee conservation and health (1).

References

1. Kasso, T.; Mackie, M.; Ramsøe, M. et al. A Queen's Tale: An Experimental Palaeoproteomic Study of a Honey Bee Queen Cell Specimen from Natural History Museum Denmark. Open Res Eur. 2025, 4, 227. DOI: 10.12688/openreseurope.18538.2
2. Crane, E. The World History of Beekeeping and Honey Hunting. Routledge, 1999.
3. Lamei, S. The Effect of Honeybee-Specific Lactic Acid Bacteria on American Foulbrood Disease of Honeybees. Doctoral dissertation. Uppsala: Sveriges Lantbruksuniv. Acta Universitatis Agriculturae Sueciae. 1652-6880; 2018; 23. https://pub.epsilon.slu.se/15460/7/Lamei_s_180509.pdf
4. Woodward, D. R. Queen Bee: Biology, Rearing and Breeding. Balclutha, 2007.
5. Kasso, T. M.; Enevold, R.; Johns, S, et al. ArcHives—Combined Palynological, Genomic and Lipid Analysis of Medieval Wax Seals. Herit Sci. 2023a; 11, 11. DOI: 10.1186/s40494-022-00848-6
6. Parejo, M.; Wragg, D.; Henriques, D. et al. Digging into the Genomic Past of Swiss Honey Bees by Whole-Genome Sequencing Museum Specimens. Genome Biol. Evol. 2020, 12 (12), 2535-2551. DOI: 10.1093/gbe/evaa188
7. Palaeoproteomics. University of Manchester website. http://www.ical.manchester.ac.uk/research/palaeoproteomics/ (accessed 2025-08-14).