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

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.