Team:UGent2 Belgium/Safety


Measures In The Lab

Wet Lab

Our experimental work is done in a GMO class 2 laboratory at the Department Biochemical and Microbial Technology at Ghent University.

The Faculty of Bioscience Engineering at Ghent University, where our lab is situated, has its faculty biosafety committee (only Dutch version of the website available). This committee makes sure the researchers are aware of the International, European and Belgian legal framework concerning biosafety. All activities of this committee take place in close consultation with, amongst others, the faculty Environment, Hygiene and Safety Committee , the faculty Ethics Committee and with some Rectoral Services.

In addition, we have committed ourselves to both the national legal and ethical requirements compliance and the international and EU regulation compliance:

- National Legal and ethical requirements compliance Environmental regulation: Decree of the Flemish Government of 6 February 2004 amending the Decree of 6 February 1991 and the Decree of 1st June 1995 “Besluit van de Vlaamse regering van 6 februari 2004 tot wijziging van het besluit van de Vlaamse regering van 6 februari 1991 houdende vaststelling van het Vlaams reglement betreffende de milieuvergunning, en van het besluit van de Vlaamse regering van 1 juni 1995 houdende algemene en sectorale bepalingen inzake milieuhygiëne” - EU regulation compliance Directive 2015/412/EC amending Directive 2001/18/EC on the deliberate release of GMOs into the environment, Directive 2009/41/EC on the contained use of genetically modified micro-organisms. Regulation (EC) 1946/2003 on transboundary movements of GMOs, Regulation (EC) 1829/2003, Regulation (EC) 1830/2003 - International regulation The Cartagena Protocol on Biosafety to the Convention on Biological Diversity (signed by the Community and its Member States in 2000. The Council concluded the Protocol on behalf of the community through the adoption of Decision 2002/628/EC) The precautionary principle - No. 15 of the Rio Declaration adopted at the 1992 UN Conference on the Human Environment and Development.

Every person who works in one of the labs of Ghent University must know and act according to specific regulations. This means that each team member signed the laboratory and workplace regulation in which he/she declares to have taken note of these regulations, to have received a specimen of these regulations and to follow the guidelines/obligations which arise from these regulations. This also includes always wearing a lab coat, safety glasses and gloves when necessary. Guidelines more specific for working with GMOs can be found in the biosafety in the laboratory document provided by the Flanders Interuniversity Institute for Biotechnology.

The hosting lab in CSB has its own Lab Manager, Gilles Velghe, who coordinates everything in the lab and is responsible for running the laboratory safely and efficiently. In addition, every member of our team got a (bio)safety and waste disposal training before performing our experiments in the lab. This training focused mainly on the safe use of ethidium bromide, which we use as a dye for DNA detection in gels. During the eventual execution of our experiments, we were supervised by our instructors, who made sure we performed everything according to the safety regulations.

Lien also helped us setting up experimental procedures and practices. These are more general cloning procedures and practices in which she has a lot of experience in. In the hosting lab, all glasswork that has been in contact with biological material is disinfected with bleach and autoclaved before it is re-used, making sure no living micro-organisms are remaining. It has an additional drying step in the oven to prevent phages from growing. When applying bleach, we wear gloves as it can cause skin irritation. Also, benches and flows are regularly cleaned with Umonium, a broad activity spectrum disinfectant. Additionally, any cultures that leave the lab are tested in order to prevent contamination of other external laboratories.

COVID-19 Measures

The COVID-19 pandemic requires particular actions to prevent further spreading of the SARS-CoV-2 virus. The COVID-19 measures are dynamic and adapted to the most recent situation. All work in the lab complies with the latest info and guidelines of Ghent University. The main rules that everyone working on campus need to follow are listed below:

Physical distancing, stay 1.5 metres apart always and everywhere.

Masks are mandatory from the moment you set foot in a building of Ghent University.

Hygiene rules with extra attention to hand hygiene. Wash hands with soap and water, hand sanitizer is present in on every bench.

Circulation plans when walking in the buildings to maintain physical distancing when moving in a building.

The University complies to the pandemic matrix (in Dutch) that is designed by all Flemish universities in collaboration with the GEES (Group of Experts regulating the Exit Strategy for the National safety council). Depending on the pandemic level, different restrictions are taken into account.

National Guidelines (in dutch) by the government are based on the guidelines of the WHO.

Project Measures

Naringenin Production

Our bacteria, Cupriavidus necator PHB-4, Escherichia coli Top10 and Escherichia coli K-12 MG1655 are common non-pathogenic laboratory bacteria belonging to risk group 1. According to this classification, it is unlikely that they will cause human or animal disease.

For the production of naringenin, these strains are genetically engineered. This brings along no additional risk as the engineered strains are merely used as 'cell factories 'in an enclosed environment. The cells themselves will not be used in applications, but only the naringenin they produce. Furthermore, the environment will not be exposed to these modified organisms, thanks to contained use and standard GLP. Nevertheless, one should be aware of the public perception related to the use of GMOs for biotechnological applications. However, it is less problematic compared to GMO plants or animals.

All experiments were carried out in a contained laboratory environment (GMO-class 2 laboratory). The E.coli strains are non-pathogenic and lab-safe strains. Transfer of the organisms occurs standard in laminar vertical flow (biosafety cabinet level 2). For the C. necator strain, we work with contained use, meaning that transfer of the organism occurs in laminar vertical flow (biosafety cabinet level 2) and additional cleaning before and after exposure to the organism.

If containment fails, however, our project poses minimal risks for the public and for the environment. Bacteria often have the capability of transferring genetic material, especially when self-transmissible vectors are used. As usually done in practice, the vectors in our experiments are not self transducible, avoiding genetic material from being easily transferred. Furthermore, SOP's for spill accidents are available in the lab.

For the upscaling the production of naringenin, the fermentor is made sterile after production, and any downstream processing would be well calculated so that only naringenin is extracted. For example, we do not want LPS in the extract; protocols are available to exclude LPS from the naringenin and testing for the presence of LPS is also possible.

Bubbly's Pearls

The use of the pearls is related to the risk of choking when not being ingested correctly or according to subscription/manual. A further possible threat is to get an overdose of our vitamin B12 /naringenin/kojibiose when abusing the product we designed.

We will try to minimize the risk by clearly stating what the recommended daily dose is and how the product should be consumed. We are not offering a drug but a supplement, hence the dosage will be lower. This gives us more margin in what the recommended dose should be, and we can keep it as low as possible (without the product losing its effect). Furthermore, a do-it-yourself principle of dosing could help minimize the risk. We will also try to make the pearl's size as feasible to ingest as possible.

Kojibiose is not yet approved for consumption by EFSA. Hence we will not use it yet in our product. We will only use it when all safety and security regulations are met.

Note: We will NOT introduce our naringenin produced in the lab in the consumables.