Royal jelly, a honey bee secretion, plays a critical role in caste determination in honey bees because it serves as the source of nutrition for young larvae destined to become queens. It is also fed to adult queens. Royal jelly possesses numerous functional properties and thus has been used as a medication, health food, and cosmetic in many countries. In this paper, we first introduce a traditional method for producing royal jelly by artificial larvae grafting and a newly developed method that does not require grafting of larvae. We describe protocols for the storage and freeze-drying of royal jelly to preserve its biological properties. Routine methods for determination of two important quality criteria, water content and trans-10-hydroxy-2-decenoic acid content, are outlined. On a dry basis, protein, carbohydrate, and fatty acids were found to be the 3 most abundant components of royal jelly. Methods for their isolation, identification, and quantification are described. Because royal jelly is susceptible to contamination with veterinary drugs and acaricides, we also describe methods for detection and quantification of some veterinary drugs and acaricides in royal jelly.Mé todos estándar para la investigació n de la jalea real de Apis mellifera La jalea real, una secreció n de abejas, desempeña un papel crítico en la determinació n de castas en la abeja melífera, ya que sirve como fuente de nutrició n para larvas jó venes destinadas a convertirse en reinas. También alimenta a las reinas adultas. La jalea real posee numerosas propiedades funcionales y por lo tanto se ha utilizado como un medicamento, alimento saludable y cosmético en muchos países. En este artículo, introducimos un método tradicional para producir jalea real mediante el injerto artificial de larvas y un método recientemente desarrollado que no requiere injerto de larvas. Describimos protocolos para el almacenamiento y la liofilizació n de la jalea real para preservar sus propiedades bioló gicas. Se describen métodos rutinarios para la determinació n de dos importantes criterios de calidad, el contenido de agua y el de ácido trans-10-hidroxi-2-decenoico. En una base seca, proteínas, carbohidratos y ácidos grasos fueron los tres componentes más abundantes de la jalea real. Se describen métodos para su aislamiento, identificació n y cuantificació n. Debido a que la jalea real es susceptible a la contaminació n con medicamentos veterinarios y acaricidas, también describimos métodos para la detecció n y cuantificació n de algunos medicamentos veterinarios y acaricidas en jalea real.

The honey bee has a well-organized system of division of labour among workers. Workers typically progress through a series of discrete behavioural castes as they age, and this has become an important case study for exploring how dynamic changes in gene expression can influence behaviour. Here we applied both digital gene expression analysis and methyl DNA immunoprecipitation analysis to nurse, forager and reverted nurse bees (nurses that have returned to the nursing state after a period spent foraging) from the same colony in order to compare the outcomes of these different forms of genomic analysis. A total of 874 and 710 significantly differentially expressed genes were identified in forager/nurse and reverted nurse/forager comparisons respectively. Of these, 229 genes exhibited reversed directions of gene expression differences between the forager/nurse and reverted nurse/forager comparisons. Using methyl-DNA immunoprecipitation combined with high-throughput sequencing (MeDIP-seq) we identified 366 and 442 significantly differentially methylated genes in forager/nurse and reverted nurse/forager comparisons respectively. Of these, 165 genes were identified as differentially methylated in both comparisons. However, very few genes were identified as both differentially expressed and differentially methylated in our comparisons of nurses and foragers. These findings confirm that changes in both gene expression and DNA methylation are involved in the nurse and forager behavioural castes, but the different analytical methods reveal quite distinct sets of candidate genes.

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