Multiplex RT-PCR with broad-range primers and an exogenous internal amplification control for the detection of honeybee viruses in bumblebees.

Bumblebees are commercially reared and transported worldwide mainly for pollination of greenhouse tomatoes. Three honeybee viruses have been reported in bumblebees: Acute bee paralysis virus, Kashmir bee virus and Deformed wing virus. We developed a multiplex RT-PCR with primers designed on highly conserved regions of the RNA-dependent RNA polymerase in order to detect a maximum range of viral variants. Rearing facilities and governmental organizations can now thoroughly screen bumblebee colonies with a cost-effective technique with an integrated internal amplification control (IAC) implementable in laboratories that strive for International Organization for Standardization (ISO) certification.

Impact of Bacillus thuringiensis strains on survival, reproduction and foraging behaviour in bumblebees (Bombus terrestris).

BACKGROUND: Bacillus thuringiensis (Bt) and its protein crystals are used worldwide, either as a spray or when expressed in transgenic crops, for the control of pest insects. However, owing to their intensive use, there exists a debate regarding the involvement of this microbial insecticide in bee colony losses. In this study, in a tiered approach using laboratory microcolonies, an evaluation was made of the potential lethal and sublethal hazards on colony reproduction and foraging behaviour of workers of the bumblebee Bombus terrestris (L.) of two commercial Bt strains: kurstaki (Dipel) and aizawai (Xentari). Bumblebees, like honey bees, are intensively used in modern agriculture for pollination and fulfil a crucial role in the natural ecosystem. RESULTS: Exposure of bumblebees dermally or via treated pollen to either of the two Bt formulations at their field recommended rates (0.1%) caused no reduction in survival. However, when applied in the feeding sugar water, aizawai killed all workers at a concentration of 0.1%, but this lethal effect was lost at 0.01%. With respect to reproductive effects, kurstaki was harmless, while aizawai at 0.1% delivered in the feeding sugar water and pollen reduced reproduction by 100 and 31% respectively. Lower doses of 0.01% aizawai in the sugar water showed no more effect. In addition, kurstaki at 0.1% and aizawai at 0.01% in the feeding sugar water did not impair the foraging behaviour, resulting in normal nest colony performance. CONCLUSION: The results with kurstaki and aizawai demonstrated that, in general, the Bt strains are safe to B. terrestris bumblebees, although in some cases there were detrimental effects that depended on strain and route of exposure. In addition, the authors believe that to draw firm conclusions regarding the hazards of Bt to bumblebees would require more information on relevant concentrations of Bt products in the environment. Hence, routine testing for lethal and sublethal effects is recommended to ascertain combined use of Bt products and bumblebees in modern agriculture.

Development of a new dispenser for microbiological control agents and evaluation of dissemination by bumblebees in greenhouse strawberries.

BACKGROUND: To date, in modern agriculture, biological control strategies are increasingly becoming the preferred pest management approach. However, the success of microbiological control agents (MCAs) largely depends on efficient dissemination into the crop. The pollinator-and-vector technology employs pollinating insects like bees for a better dissemination. In this study, a new dispenser for bumblebee workers of Bombus terrestris L. was developed. Binab-T-vector and Prestop-Mix were used as two typical MCA products for dissemination. RESULTS: In a first series of experiments in the laboratory for optimisation, the newly developed dispenser was a two-way type dispenser, 20 cm long, with two rectangular compartments and different entrance and exit holes. In addition, the amounts of MCA loaded on the workers were 10 times higher with the new dispenser as compared with the side-by-side passageway (SSP) dispenser. Typically, the highest amounts were recovered from the thorax and legs of the workers. In a second series of experiments under greenhouse conditions with the use of queen-right B. terrestris hives, successful dissemination in strawberry flowers was obtained at different distances from the hive (0-8 m, 8-18 m and 18-21 m), and the workers inoculated the first, second and third flowers that were consecutively visited. In addition, the new dispenser caused no adverse effects on worker foraging intensity, whereas a dramatic reduction was scored with an SSP dispenser. Finally, the data suggested that it is necessary to refill the newly developed dispenser at 3 day intervals. CONCLUSIONS: The results demonstrated that, with the use of the newly developed dispenser, bumblebee workers carried high amounts of MCA, and this resulted in a successful dissemination of MCA into strawberry flowers.