Abstract
Small hive beetle (SHB), Aethina tumida can feed on honey, pollen and brood in honey bee colonies. It was endemic to Africa, but since 1996 has been detected in a number of countries worldwide, including Australia, Brazil, Canada, Italy, Mexico, South Korea, Philippines and the USA where it has had economic effects on local apiculture. To improve SHB identification, we obtained the first reference sequences from the DNA barcoding 5′ COI gene region for SHB and some species of the family Nitidulidae associated with beehives. Phylogenetic analysis of SHB COI sequences (3′ COI) revealed two divergent lineages, with those from Australia and USA being genetically different from the recent detection in Italy. Many countries, including New Zealand, are currently free from SHB, and require a rapid detection method for biosecurity. Here we present the development and validation of a real-time PCR assay for detection of SHB. The assay showed high specificity and sensitivity for detecting SHB, with no cross-reaction observed with closely related species, such as A. concolor. The real-time PCR is sensitive, detecting the target sequences up to 100 copies/µL. This assay should prove a useful biosecurity tool for rapid detection of SHB worldwide.
Highlights
The family Nitidulidae comprises roughly 4,000 species worldwide[1]
DNA sequences of 5′fragment of Cytochrome Oxidase I (COI) or the Folmer region obtained in this study showed over 99% identities between the Small hive beetle (SHB) samples from Australia and USA, but they shared around 94–96% identities with the SHB from Italy
DNA barcoding of the 5′ fragment of COI region of the SHB sample from Australia and USA were highly similar with more than 99% identity (Table 2), but they are different from sequences of the Italy SHB samples (Table 2) by around 4–6% in the Folmer’s region
Summary
A number of the nitidulid beetles are associated with bee hives (Table 1), few impacts on honey bee health have been reported[1–5], and benign associations between several Brachypeplus species and honey bees have been reported. Most other associations of nitidulid beetles within beehives appear to be non-detrimental (Table 1). Another species, small hive beetle (SHB) Aethina tumida (Murray, 1867) (Coleoptera: Nitidulidae), is considered a serious pest of beehives[11,12]. The small hive beetle is native to sub-Saharan Africa, where it was originally described as a minor pest of the western honey bees[13]. Of around 30 species of Aethina known to occur worldwide[25,26, 14] of which occur in Australia[27,28], only the SHB and A. concolor Macleay are known to be associated with beehives. Further reports suggest that SHB could invade other bee species colonies, such as Austroplebeia australis Friese[35] and Bombus impatiens Cresson[2,36]
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