To take part in the iGEM competition we have taken the appropriate safety measures to keep our teammembers as safe as possible. From the moment we wanted to start conducting lab work, we asked the lab responsible person, Peggy de Graaf – Heuvelmans, to give us an introduction on the lab. She told us about the standard safety measures like washing hands leaving the lab and wearing a lab coat and safety goggles while working in the lab, as well as the devices to clean ourselves in the case of contamination.

As we started working with E. coli bacteria at first, no extra safety measures were needed. From the beginning our instructors dr. E. Dubuc and dr. S. Guo watched us executing protocols closely. This way we learned to conduct experiments properly and safely. Along the way our supervisors gave us more and more room, but of course when we were not sure about something ourselves we would ask our supervisors or other experienced people working in the lab about it. This way we learned to work with gloves with certain chemicals and when to work with the flame.

Later on we decided that we wanted to test the effectivity of our produced lysostaphin on the bacteria we had in mind, Staphylococcus aureus. Since S. aureus is classified as a biosafety level 2 organism, (1) we decided to opt for a different bacterium. During literature research we came across the fact that lysostaphin is not only effective on S. aureus, but also on its cousin: Staphylococcus epidermidis. (2) However, S. epidermids is not classified in one biosafety level unambiguously. In the U.S.A. it has biosafety level 1, (3) but in the Netherlands biosafety level 2. (4) So we spoke about this to the biological safety officer of our section, Moniek de Liefde – van Beest, who wanted us to write a risk assessment for working with S. epidermids to inform ourselves on the dangerous part of these bacteria.

In the end, we did not work with S. epidermids. We received an amazing opportunity from one of our sponsors, PAMM. As they had labs equipped for biosafety level 2 organisms, they offered us to do our experiments with S. aureus for us in their labs. This way we could take our hands off relatively more dangerous experiments than we had done before and not be at unnecessary risk.

Furthermore, we have been working with chemicals that are sometimes highly toxic, for example DMAP (toxic for the eyes and when ingested, potentially deadly for the skin (5)) for making the hydrogel. To reduce the risk of any harm done to our teammembers, we decided to only let a maximum of three people make the hydrogels, supervised by ir. R.J.E.A. Boonen. They were obliged to work with nitrile gloves and in fumehoods additionally to wearing a lab coat and safety goggles. To ensure safe handling for other experiments where we use the hydrogels, they were thoroughly washed several times after synthesis.

In conclusion, we have had no accidents in the lab that exposed any of our members to dangerous situations, thanks to everyone paying attention at instructions on safety measures and working carefully.

References

1. Emmert EAB. Perspectives Biosafety Guidelines for Handling Microorganisms in. 2013;14(1):78–83.
2. Wu J a, Kusuma C, Mond JJ, Kokai-kun JF. Lysostaphin Disrupts Staphylococcus aureus and Staphylococcus epidermidis Bio lms on Arti cial Surfaces. Society. 2003;47(11):3407–14.
3. ATCC. Staphylococcus Epidermidis (ATCC 14990) [Internet]. 2014. Available from: https://www.lgcstandards-atcc.org/products/all/14990.aspx?geo_country=nl
4. COGEM. Lijst van Pathogene Micro-organismen en agentia [Internet]. 2011. Available from: https://www.cogem.net/showdownload.cfm?objectId=650410FC-1517-64D9-CCD5324AABF10750&objectType=mark.hive.contentobjects.download.pdf
5. Sigma-aldrich Chemie BV. SIGMA-ALDRICH [Internet]. 2018. Available from: https://www.sigmaaldrich.com/MSDS/MSDS/DisplayMSDSPage.do?country=NL&language=nl&productNumber=522805&brand=ALDRICH&PageToGoToURL=https%3A%2F%2Fwww.sigmaaldrich.com%2Fcatalog%2Fproduct%2Faldrich%2F522805%3Flang%3Den