Isolation of bacterial microbiota associated to honey bees and evaluation of potential biocontrol agents of Varroa destructor

Abstract

The honey bee parasitic mite Varroa destructor is one of the main causes of depopulation of bee colonies. Bacterial symbionts associated to honey bees are known to produce a variety of bioactive molecules that have been suggested to play a protective role against honey bee pathogens. We hypothesised that among these bacteria, those colonising the external body of honey bees, and therefore able to survive and reproduce in the hive environment outside the insect gut, may be good candidate biocontrol agents to be tested against V. destructor. The aim of this study was to isolate bacterial species from healthy honey bees and dead varroa mites and to evaluate the potential miticidal effect of their spent medium containing both bacterial metabolites and viable cells, with the final objective of finding a long-lasting solution for mite control. 61 bacterial strains belonging to the Firmicutes, Proteobacteria and Actinobacteria phyla were isolated from the surface of foragers, nurse bees and larvae collected in 10 different apiaries. The most common species was Lactobacillus kunkeei (62%). Growth capability of a selection of isolates was observed at 30 and 34 °C with 1% and 20% glucose and fructose. Laboratory bioassays were conducted by spraying mites with six-day-grown bacterial cultures containing 107 cfu/ml of four strains of L. kunkeei, Bacillus thuringiensis, Bifidobacterium asteroides and an Acetobacteraceae bacterium. The effect of each strain on varroa survival was tested independently. The first three caused 95-100% mortality of mites in 3 days, indicating a potential role as natural antagonists towards varroa. The mediation of pH of the bacterial cultures did not appear to be determinant in mite inhibition, suggesting the involvement of other modes of action against varroa. The exploitation of honey bee microbiota for controlling one of the major threats for honey bee health may be a promising approach deserving further investigation.