Rapid identification of the invasive Small hive beetle (Aethina tumida) using LAMP

BACKGROUND: Small hive beetle (SHB), Aethina tumida is an invasive pest of managed honey bees and has invaded Europe from Africa. The main risk point identified for incursions of this pest into the UK is imports of package bees and queens. Surveillance of this pest, carried out by the National Bee Unit, involves monitoring imports of live bees and routine screening of sentinel apiaries around the UK through collection of hive debris for visual inspection of pests. Currently, no molecular methods are used for screening hive debris for this pest in the UK. A new invasion of this pest would likely occur at low levels and might present with partial specimens broken up in the hive or eggs from the beetle, difficult to identify by visual inspection. Therefore, we sought to develop a fast and sensitive method for detecting SHB in hive debris by developing a loop-mediated isothermal amplification (LAMP) assay.

RESULT: The LAMP assay was able to detect the SHB in 19 to 25.37 min and did not cross-react with any non-target species tested. The assay was sensitive and could detect 12pg of DNA and it was able to detect less than 1 mg of tissue in a 30 g complex matrix of honey bee hive debris. The assay could successfully amplify from crude extracts of partial tissue specimens.

CONCLUSION: This tool will allow rapid field screening of suspect beetle specimens and laboratory screening of hive debris for a notifiable exotic pest of honey bees.