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The aim of research was the assessent of the hygienic behaviour of worker bees. The hygienic behaviour was assessed by means of two test types: the traditional needle test and a test proposed by the authors that consisted in measuring the removal rate of cardboard pieces. The experiments were conducted over three seasons and comprised a total number of 60 colonies. It was proved that the cardboard removal test may replace the needle test in the selection process of bees resistant to brood diseases, but primarily in those bees that have a good expression of hygienic behaviour. The best test results are to be expected in tests which last for 24 hours.
Virgin queens were introduced into mating nuclei containing workers of different ages. The study verified whether workers younger than 8 days, comprising the younger group, are more suitable for mating nuclei than workers older than 10 days, comprising the older group. In both groups, 7 (35%) queens were lost during mating flights. The time from introduction of queens into mating nuclei to the start of egg laying in mating nuclei with younger and older workers was 13.69 and 13.73 days, respectively. Two queens in mating nuclei with older workers did not start egg laying before the 20th day when the experiment was terminated. No influence of age of workers in mating nuclei on the performance of honeybee queens was found.
In order to screen differentially expressed genes during caste differentiation induced by pollen, larvae of Apis mellifer ligustica were developed in an incubator under controlled temperature and relative humidity until emergence. Worker bee morphology was induced by addition of water-soluble extracts from pollen in larval diets. After all of the developmental periods, the external characters and developmental degree of ovaries were evaluated to confirm the caste of new adults. Transcripts from larvae of developing queens and workers were profiled by differential-display RT-PCR analysis and differentially expressed fragments were reconfirmed by dot-blot assay. Our results showed that caste differentiation was triggered by the intake of pollen extracts. Six transcript-derived fragments were isolated and three of them are reported for the first time. We conclude that differential display is a feasible approach to identifying differentially expressed genes. The identification of up-regulated caste-differentiation-related genes provided interesting clues about the activation of biochemical steps relevant to this progress.
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