There are risks in all lines of work, however those within labs must be assessed and reduced as the hazards in this environment usually have more severe consequences. In order to minimise the risks within labs there are legislations in place within iGEM, the UK and worldwide. Our lab work is in line with these legislations. As Part of our Human Practices we have looked into the national legislation for biological safety (Heath and Safety Executive). In the labs we have followed the standard safety procedures as per the regulations of the University (Occupational Health and Safety Service).
Each member of the team must abide to the policies, practices set by Newcastle University, all staff, and students are responsible for making themselves familiar with these responsibilities. Academic staff support our team to ensure that these policies and practices are followed with care. They have prior experience within the molecular biology labs we are working in. Each team member supports each other throughout the project and has received appropriate lab inductions, training in handling of chemicals, sterilisation methods, disposal and handling of contaminated material.
Every lab has safety files that are revised and modified regularly and kept up to date, the appropriate forms have been signed by every member participating in experiments. Risk assessment forms are filled out for every compound and lab experiment, and then signed off by all team members. Before entering any of the labs the Emergency Procedures were outlined by a member of each lab.
National and International Policies
The work conducted in our labs was carried out according to UK and European regulations. We reviewed these in more detail as part of our Human Practices section, to help communication within synthetic biology we also compiled a set of Synthetic Biology Communication Guidelines. Communication is a very important factor within safety as if proper practices are not carried out, due to misunderstanding, small hazards can be amplified.
Safe Project Design
Chassis (E.coli )
The chassis, which we chose, are all non-pathogenic and were selected within the design stage to minimise the risk, these were all strains of E. coli.
The strains of E. coli we used are DH5-α and BL21-DE3, these strains are within safety group 1, like most E. coli strains these present the lowest level of risk to humans. Research commissioned by HSE (Chart et al. 2000) showed that these lack the pathogenic mechanisms usually present within hazardous E. coli strains.
From the perspective of safety, aspects of our project were advantageous due to their cell free factor. Within cell-free systems random mutations and dissemination are not as likely. This utilizes cell extract for transcription and translation. Sonification and centrifugation steps remove the cell membrane. This makes them suitable for use outside of the lab, however in line with Newcastle and iGEM’s ‘Do Not Release’ policies, this was not carried out.
For the characterization of the arsenic biosensor which we produced, specific COSHH forms were required detailing the use and possible hazards involved. Arsenic is a groundwater contaminant of major significance to public health. It can get into the body via ingestion and absorption, for this reason it is not used as a fine dust as it can be breathed in which can cause serious health hazards. The health hazards related to arsenic range from irritation to internal bleeding for short term exposure. We have assessed the health risks and precautions which provide the appropriate protection with reduced exposure. The use of arsenic on the small scale that we needed was allowed within our labs as they have appropriate ventilation and fume cupboards. As arsenic is only used for characterisation longer term effects do not need to be considered. The risk team members working with this in the lab is set to as low as 'reasonably practicable' as it is with every hazardous compound.
Sarcosine oxidase was used to transform glyphosate into formaldehyde so that existing biosensors can be used to sense formaldehyde and therefore glyphosate. The concentrations produced when characterising K2205003 were detected at a minimum concentration of 10 mg/L. The testing strips used could detect a maximum value of 200 mg/L, two samples indicated this value and so greater concentrations could be present. These were 20mL cultures tested at 5mL a time. The volumes used were low, sealed tubes were used, the formaldehyde was within solution when testing the part and all experiments were carried out underneath fume hoods, hence there was no real risk to team members.
Safe Lab Work
As mentioned within the policies section, all of our lab work abided to the University regulation and Nationwide legislation and regulations. If there was anything any member of the team was unsure about they would ask other members of the team or supervisors to help or advise until they were confident. There are non-biological hazards within labs which are covered within the induction, they are similar for both labs as they are reviewed and set by university staff, complying with university standards.
All of the Labs within the university require smart card access, restricting members of the public and unqualified members access. This makes the labs a safer environment to work in and reduces the risk to the public. If biological materials required transportation between labs the appropriate policies were followed including safe storage and suitable labeling.
Sterilisation within the lab environment is particularly important as contamination and release can potentially cause a significant risk to the public and members of the lab. Sterile equipment is used and those which are taken out of their sterile packets are sterilised using the flame of a Bunsen burner or ethanol. When exiting the lab hands should be cleaned as even though gloves are worn it is important not to transfer potentially hazardous biological material to outside of the lab.
Waste was disposed of into hazardous yellow waste bins unless this was liquid waste in which case it was decanter into a waste flask which is autoclaved and disposed of. The University has its own waste disposal procedure (University Safety Policy) which another member of the lab carries out. We are required to properly seal and label these bins for the safety of others as they contain sharps and hazardous materials.
A member of staff trained in first aid is always on call and first aid boxes are also in various locations within the lab along with eyewash stations which contain instructions of use. The methods of disposal and containment of spillages and broken glassware were covered upon induction into the lab.
As some members of our team and supervisors have participated in the iGEM completion previously they were already aware of the procedures involved in shipping DNA to the registry. All of our DNA poses no real risk as we were aware of shipping restrictions within the UK and to the US when designing parts. The standard submission kit was used to send off all of the DNA requested by iGEM.