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-A hemi-nested polymerase chain reaction (PCR) was further developed for the detection of Melissococcus pluton in adult bees and honey bee products. A chloroform:isoamyl alcohol DNA extraction method was used to provide template from 154 samples of adult bee tissues, honey, pollen, whole larvae and comb cells. All 36 honey bee samples tested from a diseased colony were shown to contain M. pluton and sub-clinical infections were detected in adult bee tissues, larvae and honey (49/98; 50.0%) collected from all 9 healthy colonies from areas where EFB was endemic. All 20 adult bee tissue samples from a healthy colony from Western Australia where EFB has never been reported were negative. Of 80 bulk honey samples from six Australian states, 55 of 80 (68.8%) samples were shown to contain M. pluton whereas culture techniques detected M. pluton in 22 of 80 (27.5%) of these samples. M. pluton was detected in honey from all Australian states except Western Australia.polymerase chain reaction / Melissococcus pluton / european foulbrood / honey bees / Australia

-Four replicated experiments were conducted to determine the concentration of oxytetracycline hydrochloride (OTC) in honey bee (Apis mellifera) larvae following application of the antibiotic to honey bee colonies. In the first experiment, the mean OTC concentration was significantly greater in whole larvae than in larval guts sampled from hives on the day immediately following treatment. In two further experiments, 0.3 g, 0.5 g and 1.0 g active OTC in caster sugar was administered to single-and double-storey colonies. The mean OTC concentration was above the minimum inhibitory concentration of OTC to Melissococcus pluton for 2 to 6 days post-treatment, depending upon the dose. The daily rate of change of concentration of OTC in larvae sampled from treated colonies ranged from 0.423 to 0.672. In a fourth experiment, application of 0.3 g and 0.5 g OTC in distilled water gave equal to or higher OTC levels in larvae on the first two days post-treatment when compared to the same doses applied in caster sugar. oxytetracycline hydrochloride / antibiotic / honey bee / European foulbrood / Australia

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