Detection and Quantification ofMelissococcus PlutonInfection in Honeybee Colonies by Means of Enzyme-Linked Immunosorbent Assay

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Abstract
Enzyme-linked immunosorbent assay (ELISA) provided a rapid and extremely sensitive technique for the detection and quantification of Melissococcus pluton in honeybee (Apis mellifera) adults and larvae. The detection threshold for M. pluton cells in homogenates of adults and larvae was found to be approximately 105 cells per ml, two orders of magnitude more sensitive than microscopical techniques previously used. Samples of larvae, nurse bees and foragers from an apparently healthy colony were tested by the ELISA procedure and in some cases were found to contain up to 5 × 107 M. pluton cells per insect. These results were confirmed by haemacytometer and plate counts, and substantiate the ability of M. pluton to establish enzootic infections in apparently healthy colonies.