Safety

General Safety

Safety regulations and rules are key to ensure that what we do as scientists are safe for ourselves, colleagues and equally important to the environment and human society. Therefore, it is very important to have safety in mind from the beginning of a project. All aspect of the work has to be considered including how to handle chemicals, bacterial strains and other high risk compounds e.g snake venom and blood.

Most of the work has been performed in class 1 GMO approved laboratory according to the Danish legislation governed by the Danish Working Environment Authority [1]. The classification allows for work with GMO that posses no to low risk.

In regards to general laboratory safety rules the following was mandatory;

1. - Non authorized personnel is allowed in the laboratories
2. - Wear laboratory coats at all times
3. - No eating or drinking
4. - No cell phones

When working with GMO the main focus from a safety perspective is to detain the organisms from entering the non classified areas. Therefore, waste management and decontamination of surfaces is a vital part of safe GMO laboratory work. At the department responsible for our laboratory the waste policy states that all solid waste should be disposed of in designated biohazard waste bins. Also, liquid biowaste should be collected in designated containers and when the container is full then be handled as solid waste. When the biohazard waste bag is full it is closed and thrown in the biohazard waste bin. The biohazard waste bin itself is sealed then decontaminated with 70% ethanol and ready for destruction. All work surfaces in the laboratory is likewise decontaminated with 70% ethanol before and after every session.

Safety Around Snake Venom

The project revolves around detecting snake venom so it was apparent from the beginning that a safe work environment had to be secured, especially whenever snake venom was handled.

First of all, the snake venoms was purchased from a renowned and trusted supplier that also provided safety sheets [2]. They laid the base for the safety protocol established for working with venom in the laboratory. Venom in its solid form posse the highest risk of harming the handler as it is highly concentrated and a puff of wind may introduce the venom to the eyes or respiratory tract. As a result, only two people on the team was allowed to weigh out the lyophilized venom and it took place in a fume hood wearing gloves, dust mask and eye protection at all times. The venom was immediately dissolved in an, for the experiment conducted, appropriate buffer to minimize the above mentioned risks.

Even though venom diluted in a solution is less dangerous we still work at concentration potentially harmful to humans. Hence, experiments with venom took place in fume hoods wearing gloves and no sharp objects were used. All solutions containing snake venom was disposed off in sealed containers and afterwards sent to destruction in the same way as solid biohazard waste.

Biosafety

In order to further minimize the risk of working with GMO choice of host microorganism is an important aspect. Luckily, many laboratory strains are available and the well characterized Escherichia coli K12 DH5-alpha was chosen [3].

It has undergone several mutations that weakens its ability to compete for habitats with wild types. To mention a few important mutations, it is less efficient to adhere to the human intestine, its natural habitat, so it cannot colonize the tissue should you by accident swallow it. The strain is no longer able to transfer genes to other cells by conjugation. This is especially important as antibiotic resistant genes are often used for screening. Lastly, the strain has vital gene deletions that makes them deficient unless they are grown at media supplemented with the missing factors [4].

The project also entails work with human blood. This poses not just safety considerations for us as an iGEM team but also for the potential consumers of our detection device. Here it is important to minimize the risk of getting uncontrolled bursts of human blood due to malfunction or faulty design. Therefore, the device will operate under vacuum instead of pressure so there is less chance of the device coming apart and expose the operator to the patient's blood. In order to get human blood from a reliable source we got samples from a department at DTU that frequently works with blood and have the appropriate safety precautions in place.

For further information refer to the Final Safety Form.

References

[1] Arbejdstilsynet (2017).Klassifikation af laboratorier, anlæg til produktion mv.
[2] Latoxan (2012). Safety Data Sheet
[3] ThermoFisher (2017). Genotypes of Invitrogen™ competent cells
[4] Rayburn SR (1990). The Foundations of Laboratory Safety: A Guide for the Biomedical Laboratory Springer