Hive Safety

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In the modern world, made possible by large-scale industrial production, methods to increase the production of agricultural crops have levied an increased dependence on pollinators. These pollinators, largely the honey bee, have great disease pressure with limited management. With this in mind, our project, Varroaway, aims to develop innovative swabs for qualitative analysis of hive infestation of Varroa destructor. This project relies on the use of antibiotic resistant DH5α Eschericia coli to survive on strips and react to guanine. Containment of the resulting E. coli is essential as to not introduce antibiotic resistant bacteria outside the lab. We will further explain the details on how our project is safe for the environment and has been planned according to our institution’s regulations. Our group collaborated with the USDA-ARS here in Fargo for bee-safety practices, including how to properly don a bee suit, how to properly open a hive, and many other proper bee handling procedures.

Safe Project Design

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Our practices were done to ensure a safe handling of the modified E. coli in our laboratory and to guarantee strict adherence to iGEM’s “do not release” policy. In the future, our aim is for this project to be utilized by hobbyists and industry alike to determine Varroa destructor levels within their hives. We have planned for this practical use with existing biomanufacturing processes.

Laboratory Safety

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Despite the low health risk of our project, numerous practices are required by our institution in order to maintain a safe laboratory environment:

• Proper attire is required in the laboratory at all times which includes full-length pants and closed-toed shoes
• Personal belongings are kept outside the working laboratory
• Hands must be washed prior to leaving the laboratory
• Contaminated waste is either autoclaved or disposed of in the burn bin
• Surfaces are decontaminated upon completion of a given experiment
• Laminar flow hoods are used to avoid contamination
• Knowledge of where first aid kits, eye wash stations, and showers are located in the laboratory is required by all authorized personnel
• Knowledge of allergens is required by all personnel working with bees

Each team member participating in lab work also needs to meet these requirements:

• Complete CITI biological safety training provided through NDSU
• Be listed as a contributing member on NDSU’s project safety form
• Follow all laboratory practices outlined above

Microorganism Safety

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As mentioned previously, our project relies heavily on recombinantly producing our synthetic protein from DH5α E. coli. This strain is highly versatile in use of both plasmid amplification and protein expression and is low risk to the environment. Microorganisms are classified in biosafety levels (BSL) according to the health risk they can present to both humans and the environment.

• BSL-1 organisms are not known to cause disease in humans and they present minimal potential hazard to the environment
• BSL-2 organisms are associated with human disease and pose moderate hazards to personnel and the environment
• BSL-3 organisms may cause serious or potentially lethal disease by exposure through inhalation
• BSL-4 organisms present a high risk of aerosol transmitted infections and life-threatening disease

DH5α E. coli is classified as a BSL-1 organism by the American Type Culture Collection (ATCC) and therefore requires BSL-1 certified laboratories for handling. It also belongs to the white list set up by iGEM for use in the competition.