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Personal Safety

Safety Training

All of the students involved in our project have received safety instructions for working at the laboratory. Graduate students have attended a course offered by São Carlos Institute of Physics (IFSC/USP) about basic safety aspects, and then instructed undergraduate students about the precautions to be taken. The course included topics such as: the use of appropriated clothing and personal protective equipment; correct use of equipments such as autoclave, laminar hood flow, electrophoresis chamber and centrifuge; handling, safety aspects and disposal of biological and chemical agents; specific danger of the main reagents used at the lab; precautions about bioaerosol.

Risks of the Project

Our team’s project does not present any major risks for the safety of anyone working directly or around it. As our project only involve bacteria that present low health risk, we used a BSL 1 laboratory for the development of the project.

• Although Pseudomonas aeruginosa is an opportunist pathogen (Risk group 2), we did not handle the bacterium but only its DNA parts.
• Escherichia coli can cause health issues (irritation to eyes, skin, respiratory and gastrointestinal tract), but if it is handled with precaution, the risk is minimal. All of our students have attended safety training and are applying their knowledge at the lab.
• Bacillus subtilis is a non-pathogenic bacteria and, therefore, does not present any health issues.
• The DNA parts used in the project do not present any major risks.

Precautions Taken to Avoid Unexpected Release of the GMO

Disposal of Biological Material at the Laboratory

We were very careful in the disposal of biological material at the laboratory. All liquid cell cultures were autoclaved and, after reaching room temperature, disposed on the sink. Used agar plates were autoclaved inside a bag and disposed on common waste. Pipette tips were incinerated before being disposed. There was also the possibility of using a powder disinfectant called Virkon. Usually, Virkon is mixed with the contaminated material for 30 minutes. After that, the materials were discarded on common waste.

Safety in the Design

In order to minimize the risks of unwanted release of the developed bacteria, we designed a disposal mechanism in our final device. The modified bacteria are physically contained in a chamber and a device that releases disinfectants agents (such as sodium hypochlorite) was designed to be activated after the detector kit has been used. In this way, the chance of release of the developed organism by the public is minimized.

Risks to the safety and health of the general public

We were able to identify two main risks in this case:

• Escherichia coli could infect people by being present in the consumed water or food. In this case, the individual would probably suffer a gastrointestinal tract irritation. It would not be any different from an infection caused by E. coli originated from animal feces.
• The plasmids with biobricks contain resistance related genes. If these genes were transmitted to pathogenic bacteria, they could cause harm to the general public.

Risks to the environment

As we use inducible promoters, if the trigger reactant isn’t present our genic circuit is off. But even if it were on, it would not cause any damage – our genetically modified bacteria do not release any substance; it only turns to a different color under determinate circumstances. Besides that, both E. coli and B. subitilis are of common occurrence in the nature. Therefore, no risks to the environment were detected.

Risks to security through malicious misuse by individuals, groups, or countries

We did not identify any major risks in our project for malicious misuse. None of the used parts has any direct relation with virulence and the organism is a non-pathogenic bacteria. Although qteE affects the expression of virulence factors in P. aeruginosa, it has been found that a higher qteE concentration leads to the decrease of virulence. Therefore, qteE does not present risks through malicious misuse (http://jmm.sgmjournals.org/content/60/1/22.full.pdf).

Risks that might arise if the project would be used as a commercial/industrial product

The main risk arising from our project’s growth would be the detection kit disposal – as it would contain the genetically modified bacteria. In order to minimize this, a disposal device was projected in our final device, as mentioned above (Safety in the Design).

Institutional Biosafety Committee and changes made to the project based on their review

Here we have an Institutional Biosafety Committee named CIBio IFSC. All the procedures applied in our team’s project have been discussed with the members. In the initial planning of our project, Professor Ana Paula Ulian de Araújo has shown concern about us extracting and using Streptococcus pneumoniae DNA – which can be the cause of serious disease in humans, such as pneumonia and meningitis (risk group 2). After discussing that subject, we’ve adapted our initial project not to use S. pneumoniae anymore.

Our Safety Forms

• About Our Lab
• Safety Form
• Safety Spreadsheet