Team:Virginia/Safety




Safety



Organisms

As we designed our experiment, choosing a chassis organism was fundamental to how we were going to accomplish our project. By choosing Paracoccus denitrificans, we chose a chassis organism that is currently implemented in several wastewater facilities across the country, including the local wastewater facility near us in Charlottesville, Virginia. Pc. denitrificans is a non-pathogenic chassis, and the genetic circuit we are implementing does not harm humans, animals, or plants in any way. It is a risk group 1 organism and is non-pathogenic. We ensured that all parts we are using come from Nitrosomonas europaea and Escherichia coli (K12 strain), both risk group 1 organisms. Furthermore, after we spoke with the University of Virginia’s Aaron Mills, we worked with two different promoter parts that could control the level of expression. This idea was actually adapted from the more mechanical control mechanisms used in the local wastewater treatment plant, which was subsequently integrated into our main circulatory. Furthermore, we opted not to incorporate an “kill-switch” into our final composite circuit, as according to Professor Mills, similar projects, like Anammox were designed with Kill-Switches, which were impractical for wastewater treatments plants to regulate.


Containment

Our device is designed to be implemented in a wastewater facility. Currently, multi-cultures are used in wastewater facilities, so containment for the bacteria has already been designed into the facilities. In most facilities, there are two methods to remove the bacteria from the cleaned water. The first method uses chlorine. One part of the facility will insert chlorine into the processed water, oxidizing cellular material of the bacteria. This procedure is often used because it is cost-effective and well-established. Another method of removal uses UV light. UV light prevents the bacteria from reproducing by damaging the DNA and RNA of the bacteria. This procedure is often used because it leaves no harmful residue in the water and is highly effective.


Our Lab and Lab Procedures

Safety is always the number one concern of the iGEM Lab. The appropriate PPE was worn at all times for all facets of the experimentation. We would follow typical disposal methods were followed: all liquids were bleached before being disposed off, all solid waste were put in the appropriate waste containers, and when applicable broken glassware was properly disposed of in a proper broken glass waste container. Any time eye washes/chemical showers were used, incidents were properly documented and the proper staff was notified. All glassware was sterilized after usage. We are working in a Biosafety level 2 lab.