Laboratory

The Wageningen University and Research centre biosafety policy focuses on working with biological materials in a way that is safe for both human beings and the environment. Biological materials include genetically modified organisms (GMO), biological agents, plant pathogens/quarantine material and animal by-products. The spread of these materials needs to be restricted or limited when they are directly hazardous to humans, animals or plants, but also for economic and/or ethical reasons. In all cases, the researcher, lecturer, student or third party is consciously working with material that may have hazardous effects of which the person is not always aware in advance. In such cases it is necessary to ask advice of the biosafety officer in order to be able to work safely with the material and in order to meet applicable laws and regulations.

In the Netherlands permits for working with GMOs are handled by the following institues or persons:

• Permit issued by: Dutch Ministry of Infrastructure and the Environment
  
• Carried out by: Dutch Bureau for Genetically Modified Organisms
  
• Contact persons: Wageningen UR biosafety officers

The Laboratory of Microbiology at Wageningen UR follows rules that allow the handling of genetically modified organisms that are approved and accepted by European Union. To prevent the contamination of the environment, waste is separated into biological and regular waste. Liquid biological waste is contained into bottles and autoclaved. After autoclaving, the liquid from the bottles is emptied in the sink. Before disposing the waste that was exposed to microorganisms it has to be autoclaved. Loose plastic containers (tubes, petridishes) are collected in special metal bins with heat resistant bags within the laboratory.

Every team member from BananaGuard group participated in a general tour around the laboratories, received safety instructions and had a short exam about safety procedures. Within those training sessions a student obtains the knowledge on how to safely work in the university's ML-1 laboratories. ML-1 is low-risk containment level of laboratory that is appropriate for work with non-pathogenic genetically modified organisms, while the highest level, ML-4, stands for high-risk containment level of laboratory and is appropriate for highly pathogenic GMOs.

Additionally, we received a lab journal and a book named "A practical guide to the lab". The book includes all necessary information for daily work in the laboratory. It includes topics such as safety, waste disposal and description of methods such as DNA isolation, PCR, transformation and sequencing. The booklet's purpose is to provide a clear perspective as to how experimental approaches can be implemented in a safe and sustainable way. During the safety tour around the laboratory we were informed about lab rules, how to react in case of an emergency, the location of fire extinguishers and safety equipment. The safety trainer also provided information about waste disposal (especially about waste containing GMOs) and the storage of laboratory equipment. During the tour the trainer explicitly stressed the usage of good laboratory practice (GLP). The safety lecture, which differs from the safety tour, covered how to prevent emergencies and how to take action in case an accident happens.

The safety lecture and lab tour are fundamental for preventing possible risks to the safety and the health of team members. Safety instructions are given to avoid dangerous situations at work. While working in the laboratory, some basics have to be taken into account. A person has to behave according to the safety rules (washing hands, cleaning working space) and has to wear basic laboratory equipment. The basic laboratory equipment includes laboratory coat, protection glasses and special rubber gloves which protect against corrosive chemicals. To implement GLP a person should not wear shorts, open shoes and has to be careful with open gas flame. If working with hazardous chemicals (gel preparation, cleaning agents, etc.) basic laboratory equipment is required and work in the fume hoods. Chemicals are appropriately labelled with names and signs if they are dangerous. To minimize the risk to the health of the team members also waste disposal is carefully separated and labelled. For instance, to prevent a person from cutting himself/herself, used sharp tools are disposed of in special yellow plastic containers.

In a case an accident happens, despite the aforementioned precautions, there is emergency equipment. In the laboratory there is one eyewash station. Outside of the laboratory (divided by the fireproof doors) there is an emergency shower in case a person is on fire or spilled with acid. In the building, including hallways and laboratories, there are many reachable and working foam fire extinguishers, and also fire blankets. Every year Wageningen University organizes several fire alarm exercises. In order to prevent emergency situations, every team member needs to follow the instructions we got at the safety lecture. If we behave according to the safety rules, we can prevent many so called almost-accidents.

To test the effectiveness of our final product and intermediates, we conducted experiments with Fusarium oxysporum. This organism, which requires special protocols, was not used in the ML-1 lab but in a ML-2 lab at a different location. Work in this laboratory happened under strict supervision and according to their safety protocols.

Organisms expressing different antifungals were tested for their functionality as a biological soil control agent for banana plants in special compartment of greenhouse. The mentioned compartment was used for genetically modified fungi. Therefore the safety practice is the same as genetically modified chassis Pseudomonas putida. All individuals involved in the activities during plant maintenance and scoring used disposable clothing (coats, gloves, shoe covers) which were discarded in double red plastic bags for autoclaving after completion of tasks. Remaining plants, pots, soil, stems, foliage, used tools were discarded in double red plastic bags and inactivated by autoclaving. The permission for using GMO organisms with plants in the special compartment in the greenhouse had to be temporarily extended with detailed information about the constructs and organism. The detailed protocol was submitted to safety officer in the greenhouse and approved.

For more information regarding safety in our project read the Safety form.

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