SAFETY

There is a lot of dangers in a laboratory. So, everything cannot be done and there are rules to respect to protect the environment and ourselves. Working with GMOs also requires a strict safety policy for obvious ethical reasons.

First, we will present here how the safety department in INSA is organized: what kind of structure received us, what is the French legislation concerning risks and how we have been formed. Then, we will detail what kind of risks we could met during the lab work and how we should avoid them with good laboratory habits. Finely, we will explain how we constructed our project to minimize the health and safety issues.

INSA Safety Department

Organization of the INSA Safety Department

We have been received by the LISBP. The head of the laboratory where we worked is Carole Jouve and the organization of the safety department is described in the figure 1 below. We are very grateful for the welcoming of our colleagues and the time they spend to form us.

organization chart — **Figure 1:** *Organization chart of the INSA safety department.*

Legislation and French labor Law

We are working in a french engineering school (INSA Toulouse), thus we have to respect the french national regulations about working conditions and manipulation of GMOs.

As we work with microorganisms, we are concerned by the regulation on workers protection against risks resulting from their exposure to pathogenic biological agents (Decree No. 94-352 of 4 May 1994). It also includes human endoparasites which may cause infections, allergies or toxicity. This Decree is the French transposition of the Directive 90/679 / EEC and is also transcribed in the Labour Code (Articles L4421-1 R4421-1 to R4427-5). This Decree of the 16th July 2007 describes the technical preventive measures that must be followed in research laboratories, where workers are likely to be exposed to biological pathogens. We must obey to the rules of health, safety, and preventive medicine applied in public services in France (Decree No. 82-453). This decree refers to the Labour Code, Public Health Code and Environmental Code.

Formation received

To follow the legislation, we have been formed by Nathalie Doubrovine, the supervisor of the safety department of the LISBP. Her goal is to ensure both the well-being of the employees and the respect of safety rules and risk prevention. During the summer, we received different formations:

General information: two formations have been done, one during a meeting with Nathalie Doubrovine and the other with the software NEO. General information has been presented about the chemical, biological and fire risks. We also learned the prevention of those risks and the reaction to have when one of those risks occurs. At the end of the formation, a little test was done to see if everyone understood everything. Also, before using any new device, a quick formation is done.
Autoclave formation: our team has attended an autoclave training, which showed us the explosive and implosive dangers of the dispositive and the security measures to take to protect ourselves. A lab coat, heat resistant gloves and glasses were required for the manipulation of the autoclave. The standard protocols of loading and unloading it were also demonstrated.
Ethidium bromide room (Figure 2): because of the oncogenic risk of the ethidium bromide, we have a specific room where we must work with this solution. Nothing can get outside the room when it comes inside and everyone in the room must wear protection gloves, a lab coat and protection goggles. UV are also used in this room and protection mask must be wore during its use.

A generic square placeholder image with rounded corners in a figure. — **Figure 2:** *Ethidium Bromide Handling Room.*

Knowing the risks and minimize them

Hazards we could met

In the laboratory, there is a large variety of risks we could met. We will detail what risks could occurs and how the LISBP managed them here.

Main hazards:

External risks (earthquake, security issues…):

LISBP is concerned by many different types of hazards due to a wide spectrum of scientific activities. One hazard category gathers risks that are not directly linked to the LISBP research activity such as the risks due to working conditions (noise, thermal atmosphere, etc.), electrical hazards, work on computer screens, falls, etc. The main hazard categories concern the substances handled for research purposes such as class 1 microorganisms (GMO or not), urban wastewaters, chemical products among including CMRs, cryogenic fluids, etc… A second main hazard category relates to equipment and pilot plants with specific risks such as equipment using pressurized liquids, gases or steam (autoclaves), instruments generating non-ionizing radiations (Laser, UV lights) and electromagnetic radiations (RMN).

Hazardous chemicals:

List of hazardous, chemicals & inventory:

The list of hazardous, chemicals and inventory is managed independently by the each LISBP sub group. A common LISBP list of CMR substances is monitored and updated.

Waste management plan:

The chemical waste is managed in accordance with the environmental code. First, the waste is stored on a specially adapted LISBP/INSA site, before being disposed by a subcontracted service provider in accordance with the ADR rules concerning the waste treatment, valorization or destruction. Each waste removal is accompanied by tracking form for monitoring of hazardous waste.

Vacuum/pressurized equipment:

Pressurized equipment is monitored according to the Decree from 15 March 2000 concerning the work with the vacuum and high-pressure equipment.

Cryogenic/high temperature equipment:

Implementation of adequate means of prevention.

Biological safety:

Biological safety plan:

The laboratory applies the measures states in the articles of French Labour Code: L.4421-1 et R. 4421-1 a R. 4427-5 (Directive 90/619 on life insurance from 26 November 1990).

Biological material Type of organism used:

Prokaryotes, unicellular eukaryotes. Risk group: Risk group 1 Origin: E. coli and yeast mainly. Other biological: DNA, RNA... Facilities/Equipment Biosafety level: The majority of the LISBP facility is operated at Biosafety Level 1.

Biosafety cabinets:

The use of PSM type II guarantee the sterility of the manipulation and the protection of the environment and of the manipulator.

Decontamination plan:

Certain laboratory rooms are subjected to gaseous decontamination once per year or upon request.

Biological waste management plan:

The biological waste (liquid or solid) is pretreated chemically (bleach) or by autoclaving. Glass, sharp or jagged objects are disposed via a specialized company (DASRI).

Biological emergency plan (exposure spill):

In case of exposure spill, the liquids are absorbed through suitable means and further treated as biological waste.

Good laboratory practices

We have received a formation about the safety in the lab that teaches us some basic rules. Those rules are listed in the summary diagram below.

Safety and project construction

Safety management, french legislation and ethics took an important part in the way we managed our project. The brainstorming have been particularly affected by those points. Indeed, we wanted to work on a project that can live outside the iGEM competition and outside the lab. So, very soon, we select our ideas depending on “will the use of GMOs become an issue for the safety of the customer by also for the environment”. Our project Cerberus need GMOs but we develop it to avoid any contact with the user of the product but also with the environnement. At the end, we produce a protein in laboratory with GMOs but only the protein, and not the organism that produced it, is used.

If this part of the construction of our project interest you, we invite you to read the Human Practices section.

No dogs were harmed over the course of this iGEM project.

The whole Toulouse INSA-UPS team wants to thank our sponsors, especially:

And many more. For futher information about our sponsors, please consult our Sponsors page.

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