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Nosema spp. spores are extremely resistant to external stress factors and can survive several years without losing the ability for further infection within the insect body. For this reason, combating nosemosis is difficult. Some beekeepers add ethanol to the sucrose solution before the winter to prevent nosemosis infection and to cure already infected colonies. Others feed infected colonies with herbal ethanol extracts. Therefore the aim of this study was to evaluate the ethanol impact on bees infected with Nosema spp. Four groups of uninfected and Nosema spp. infected bees were fed with sugar-water syrup (1:1) supplemented with ethanol at the following concentrations: 10%, 5% and 2.5% and 0% as a control (only a sucrose syrup). Generally, bees consumed 10% EtOH solution in an amount even 50% lower than in other concentrations. The impact of EtOH on the increase of bees’ mortality was observed at a 10% EtOH concentration for healthy bees and even from 5% EtOH concentration for Nosema spp. infected bees. In our study the highest number of Nosema spp. infestation was noticed for bees fed with 5% EtOH and the lowest pH level was also measured for this group of bees. Therefore, a clear correlation was observed between the feeding bees with EtOH, which resulted in the acidification of bees, and the degree of Nosema spp. infestation. A synergistic effect of the ethanol and nosemosis on the rise of the mortality of bees has been observed. The addition of ethanol to sucrose syrup facilitates conditions for the development of nosemosis in honey bees. The strongest effect of ethanol on the level of Noseama spp. infection was observed for the 5% ethanol solutions. Moreover, ethanol at 10% concentration in sugar syrup exerts severe toxic effects even on healthy bees. All these factors induced immune-suppression in bees and enhanced the level of Nosema spp. infestation.
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.
In order to perform a phylogenetic analysis of Nosema spp. spores, samples of 10 worker bees each were collected from 30 infected colonies (the presence of spores was confirmed by light microscopy) kept in 15 apiaries in north-eastern Poland. Both N. apis and N. ceranae are common in this region (mixed infection N. apis/ N. ceranae – 60%, N. ceranae – 37%, N. apis – 3%). The DNA samples of N. apis were 100% identical with the N. apis sequences deposited in the GenBank database in Queensland, Australia, Spain, New Zealand, Lithuania and Tasmania in Australia. The DNA samples of N. ceranae were found to be 99.5%-100% identical with the N. ceranae sequences previously published in Italy, Germany, Switzerland and Austria.
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