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The aim of this study was to determine which method of hygienic behaviour assessment is more reliable: the evaluation of the pierced brood removal rate or the evaluation of the freeze-killed brood removal rate. Additionally, we aimed to determine whether freeze-killed brood should be placed in colonies when defrosted or still frozen. Defrosted freeze-killed brood was removed faster within a 24 h period. The removal rates for pierced brood and frozen freeze-killed brood were similar in hygienic colonies. In non-hygienic colonies, pierced brood was removed at a significantly slower rate than frozen or defrosted freeze-killed brood. The mechanisms of removing frozen and defrosted freeze-killed brood were similar to each other and different from those observed in the case of pierced brood. The defrosting of brood prior to its introduction into colonies seems inadvisable, as it accelerates brood removal. Our results confirm the hypotheses of those researchers who believe that the frozen freeze-killed brood removal test is not always appropriate. A good solution is, therefore, to perform the frozen freeze-killed brood and pin-killed brood removal tests simultaneously. The time from the beginning of the tests to the moment 50% and 75% of dead-brood cells have been cleaned up should be assumed as the appropriate duration of the hygienic behavior evaluation tests.
Intestinal microflora is a very important part of the digestive system in every animal, and plays a role in the synthesis of vitamins and the metabolism of many toxic chemical compounds. The indigenous intestinal flora of bees changes even as a result of changing their diet from natural to artificial or placing them in cages. Such factors have an impact on the health of bees and on the strength of whole colonies. In our study, intestinal fungi isolated from healthy bees and from bees infected with Nosema spp. belonged to two genera: Candida and Saccharomyces. The approximate numbers of yeast CFUs (colony forming units) obtained from healthy Apis mellifera carnica and Buckfast bees were, respectively, 2880-5180 and 1056-4120. Apis mellifera carnica and Buckfast bees were similarly sensitive to slight Nosema spp. infections, but heavy infestations had a greater impact on the intestinal microflora of A. m. carnica. The degree of Nosema spp. infestation had an impact on the quantitative composition of the intestinal microflora of bees. Slightly infected bees of Apis mellifera carnica had up to 44 915 yeast CFUs per bee, and Buckfast bees up to 28 705 yeast CFUs per bee. Surprisingly, a heavy infestation reduced the number of yeast CFUs to no more than 120 in A. m. carnica bees and to no more than 164 in Buckfast bees. Therefore, in studies in which the number of yeast CFUs is used as the main indicator of stress in bees, the potential presence and the degree of Nosema spp. infestation needs to be taken into account.
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