Apidologie 34 (2003) 19-27
DOI: 10.1051/apido:2002047

The detection of Melissococcus pluton in honey bees (Apis mellifera) and their products using a hemi-nested PCR

Ben Alexander McKee^a, Steven Philip Djordjevic^b, Russell David Goodman^c and Michael Alan Hornitzky<sup>d

a</sup>  The University of Melbourne, Institute of Land and Food Resources, Parkville, Victoria, Australia, 3052
^b  New South Wales Agriculture, Elizabeth Macarthur Agricultural Institute, Camden, New South Wales, Australia, 2570
^c  Department of Natural Resources and Environment, Institute for Horticultural Development, Victoria, Australia, 3176
^d  NSW Agriculture, Elizabeth Macarthur Agricultural Institute, PMB 8, Camden, New South Wales, Australia, 2570

(Received 24 August 2001; revised 5 February 2002; accepted 6 May 2002)

Abstract
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.

Key words: polymerase chain reaction / Melissococcus pluton / european foulbrood / honey bees / Australia

Correspondence and reprints: Michael Alan Hornitzky
    e-mail: michael.hornitzky@agric.nsw.gov.au

© INRA, EDP Sciences, DIB, AGIB 2003