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Investigating novel control methods for honey bee pests and diseases - PH0505

This project will consist of four main modules with different themes in support of bee health policy.
1 . Microsatellite markers and European foul brood resistance
This module proposes to catalogue honey bee races in the UK and to determine which honey bees exhibit EFB disease resistance. Initially, genetic stock will be sourced to represent the different races of honey bees. Each race will be characterised using microsatellite markers which already exist in the literature. Honey bees from England and Wales will be then be catergorised using the same markers to determine the genetic diversity present. To investigate resistance, candidate apiaries will be identified by having never shown EFB symptoms despite being in a high risk area. These apparently resistant bees will be characterised to determine the relatedness of the colonies to the known races. Such data may indicate a possible genetic link to EFB resistance.

2. Alternative non-chemical approaches for the control of Mellisococcus plutonius (Mp)
This module will investigate the use of bacteriophage and bacterial antagonists as alternative non-chemical control methods of Mp. Bacteriophage (also known as phage) are viruses which infect bacteria and reproduce at the expense of the host cell’s own metabolism. Lytic phage kill their host by inducing lysis, and phage particles thus released then infect other susceptible cells in the vicinity. This ability, along with the high degree of host specificity exhibited by phage, lend them great potential as biological control agents, and many examples of their potential have been reported (McKenna et al., 2001; Park et al., 2000; Peek and Reddy, 2006). Phage offer a novel approach for the control of bacterial pathogens of the honey bee since introduction of the control agent into hives would be relatively simple and dispersal of phage throughout the hive would be done during the normal action of worker bees. CSL staff have considerable experience working with bacteriophage and propose to identify phage which have an action against Mp. T-RFLP is a powerful method for the identification of bacterial species present in environmental samples. The technique involves amplification of conserved bacterial DNA sequences using polymerase chain reaction (PCR) primers labelled with fluorescent markers and digestion of the amplification products with a defined set of restriction enzymes and detection of the resulting fragment sizes by capillary electrophoresis. Only those fragments labelled with fluorescent markers (i.e. the terminal fragments) are detected, thus providing a way to simplify the otherwise highly complex pattern of fragments obtained from bacterial populations. The method can identify taxa present within the sample, and as such is more powerful than other community fingerprinting methods which either do not allow any level of identification or may require prior knowledge of the organisms present. We propose to use this method to screen bee gut microflora in infected and uninfected larvae to identify potential antagonists.

3. Use of fusion proteins for control of the small hive beetle (SHB)
The SHB represents a major threat to the long-term sustainability and economic prosperity of EU and UK apiculture if it is imported into these regions. Currently SHB is a notifiable pest and contingency measures exist for the eradication of SHB should it be found in the UK. However, control measures for SHB are limited and their use has given variable results. The current small hive beetle project (PH0503) is on target to identify novel attractive compounds that could be used in lures for the early detection of SHB. This project has also shown preliminary evidence for a novel control method based on the use of a fusion protein. Fusion protein technology has been patented and developed for the production of novel environmentally benign pesticides jointly by CSL and the University of Durham (Application No. PCT/GB2002/003598). Novel insecticidal fusion proteins have shown potential for the control of lepidopteran (Fitches et al., 2002; Fitches et al., 2004; Trung et al., 2006), homopteran (Down et al., 2006; Trung et al., 2006), dipteran and coleopteran (Fitches, personal communication) pests. This module proposes to expand efficacy testing of these compounds for action against SHB. If successful, it is anticipated that this work would result in a novel lure and kill system for SHB control.

4. Monitoring the impact of Nosema ceranae
Experiments will be conducted to quantify the impact of Nosema ceranae on honey bee colonies in the UK. Apiary sites shown to contain N. ceranae in 2007 will be identified and resampled. Colony loss data will be collected and the level of the pathogen quantified using real-time PCR. Apiaries testing positive for Nosema apis will also be sampled for a comparison of pathogenicity. Also, treatment of Nosema will be investigated.
1.1 Source various stocks of honey bee stock and characterise using microsatellite markers and genetic analysis.

1.2. Identify EFB resistant and susceptible colonies and assess the genetic relatedness of the colonies in the identified apiaries to the brood stock characterised in 1.1.

2.1 Isolate phage lytic on Mp

2.2. Investigate the ability of phage to limit the growth of Mp in vitro and in honeybee larvae

2.3. Develop T-RFLP fingerprinting protocols for characterisation of microbial communities in honeybees.
3.1. Produce fusion proteins in sufficient quantity for use in laboratory bioassays.
3.2. Investigate the potential of fusion proteins to act as a novel control agent for SHB.
3.3. Examine the potential for a lure and kill system for SHB control.

3.4. Maintain the laboratory cultures of the small hive beetle (SHB) in CSL’s licensed quarantine facilities and develop techniques for provision of insects at appropriate developmental stages for use in laboratory bioassays.

4.1 Assess the impact of Nosema ceranae and Nosema apis on colonies which tested positive for these pathogens in 2007.

4.2 Assess the biology of and the treatment efficacy for both Nosema species
Project Documents
• Final Report : PH0505 Bee Health - Investigating novel control methods for honey bee pests and diseases   (615k)
Time-Scale and Cost
From: 2008

To: 2009

Cost: £130,000
Contractor / Funded Organisations
Central Science Laboratory
Bee Diseases              
Bee Health              
Plants and Animals              
Fields of Study
Plant Health