Safety and Security

iGEM is Staying Out of Danger.

This section provides an overview of the safety and security measures implemented to ensure responsible research and protect both the team and the community.

Our lab

Figure 1.
Figure 1. Photographs of the laboratory, including the benches and the laminar flow hood, essential for the sterility conditions required for our project.

The wet lab team spent two months at the Translational Synthetic Biology Lab, led by Dr. Marc Güell Cargol Marc Güell Cargol Popup Image Marc Güell research work focus on leveraging new gene editing technologies for therapeutic purposes. He is a well-known researcher in his field. , and the Synthetic Biology for Biomedical Applications Lab, led by Dr. Javier Macia Santamaria Javier Macia Santamaria. Popup Image Javier Macia research work focuses on engineering microorganisms to transform waste into valuable products, such as biodegradable plastics and renewable fuels like butanol and hydrogen.
Both of these labs are part of the Universitat Pompeu Fabra and are located within the PRBB building in Barcelona. Before starting any lab work, our instructors and principal investigators (PIs) provided us with lectures on lab safety, general rules, and potential hazardous situations, ensuring we were well-prepared for the laboratory environment.

Safe laboratory work

The lab we worked in is a Biosafety Level 1 (BSL1) lab, designed for work involving well-characterized agents that are not known to consistently cause disease in healthy adult humans and present minimal potential hazard to laboratory personnel and the environment (see Figure 1). BSL1 labs are equipped with standard microbiological practices, such as restricted access during active experiments, the use of Personal Protective Equipment (PPE), and protocols for decontaminating surfaces and materials. The laboratory is also equipped with essential safety features like handwashing facilities, readily available disinfectants, and proper waste disposal systems to prevent contamination or accidental exposure.
Our project strictly adhered to these safety protocols to ensure a secure and compliant laboratory environment. We worked exclusively with nonpathogenic organisms suitable for BSL1 conditions, which involved handling agents that pose minimal risk. No work was conducted with organisms requiring Biosafety Level 2 (BSL2) or higher containment, ensuring minimal risk to human health and the environment. This precautionary approach allowed us to focus on the research while prioritizing the safety and security of all team members.
Our team had access to a dedicated lab bench and shared other spaces with the lab members, such as benches with Bunsen burners for maintaining sterile conditions, air flow cabins for contamination control, incubators for growing cultures, and plate readers for measuring absorbance or fluorescence in assays. To ensure safe and respectful procedures when moving between areas and sharing materials, we collaborated closely with other lab members, fostering a cooperative and efficient working environment.
For detailed information on specific lab security protocols, such as evacuation procedures, lab safety rules, and sample transfer guidelines, downloadable documentation is available below. These documents provide comprehensive guidelines to ensure the safety and security of all lab personal and the integrity of the research conducted.

Safety Protocols and Reagent Selection

The Escherichia coli strains used in our experiments were competent NZY 5α; and EC24 cells, which are not pathogenic. Similarly, the Cutibacterium acnes strain KPA171202, used in our laboratory, is not pathogenic (1, 2).
Therefore, the primary safety precautions include wearing gloves, lab coats, and lab glasses to prevent skin or eye contact and maintain culture sterility. One important aspect to highlight is that gloves must be removed when working near the Bunsen burner, as wearing plastic gloves increases the risk of burns. Additionally, it is crucial to follow proper waste disposal protocols for all biological materials to prevent their accidental release or contamination. The cultures and consumables were autoclaved before disposal and regular decontamination of work surfaces with appropriate disinfectants were mandatory to maintain a clean and safe work environment (3). Detailed protocols for handling spills and exposure were reviewed and practiced to ensure prompt and effective responses in the case of an accident. Adherence to these safety in compliance with iGEM recommendations, this section presents a succinct overview of the safety protocols that were employed in our project. Measures ensured that our work with these nonpathogenic strains was conducted safely and efficiently.
When selecting reagents for our experiments, we prioritized safety and sought to avoid those known to be hazardous. Recognizing our limited experience in laboratory work, we consciously chose to use SYBR Safe instead of Ethidium bromide (4). Both of these reagents perform the same function in visualizing DNA in PCR gels; however, SYBR Safe is a significantly safer alternative. Ethidium bromide, while effective, is a potent mutagen and carcinogen, posing serious health risks upon exposure. In contrast, SYBR Safe offers comparable sensitivity for DNA detection but with greatly reduced toxicity, making it safer for both the user and the environment. Additionally, SYBR Safe can be disposed of more easily and safely, minimizing the environmental impact of our laboratory activities. This decision reflects our commitment to maintaining a safe working environment while achieving reliable results in our experiments.

Future Risk

As we look toward the future development of our project, the potential application of our engineered Cutibacterium acnes strain as a treatment for scabies presents both opportunities and challenges. The ultimate goal is to create a topical lotion that can effectively treat scabies, a prevalent skin disease that affects public health worldwide. This lotion would be designed not only for infected individuals, but also as a preventive measure for those at risk, with the expectation of minimal or no side effects.
However, as with any medical treatment, the development and use of this product raises important ethical and safety considerations that need to be carefully evaluated.

Regulatory Compliance

Since our engineered C. acnes strain could be used in a medical context, it would need to undergo rigorous testing and meet a series of regulations to ensure patient safety. The product would be classified as a medical treatment, which means it would need to comply with guidelines established by regulatory bodies, such as the European Medicines Agency (EMA).
To meet these requirements, the lotion must undergo a series of preclinical and clinical trials. Initially, tests on animal models were necessary to evaluate the safety and efficacy of the treatment. These tests would help to identify any potential side effects, such as allergic reactions or other adverse effects on the skin microbiome. After successful animal testing, clinical trials will be conducted on human subjects to further assess the safety and effectiveness of the lotion under real-world conditions.
While testing on animals and humans is necessary to ensure the safety and efficacy of treatment, it also raises ethical concerns. The use of animal testing, in particular, must be carefully justified, and alternative testing methods should be explored whenever possible to minimize harm to animals.

Unintended Consequences

One of the key risks associated with the use of our lotion is its potential for unintended side effects. Despite efforts to ensure the safety of the product, there is always the possibility that some individuals may experience allergic reactions or other adverse effects. For example, the engineered C. acnes strain can alter the delicate balance of the skin microbiome, leading to unforeseen complications. Although the strain is designed to be safe and nonpathogenic, further testing will be required to monitor its long-term effects on skin health.
Additionally, the engineered bacteria could interact with other microbial communities on the skin in ways that are not yet fully understood. Although C. acnes is adapted to specific human-associated niches and is not expected to thrive outside these environments, careful consideration must be given to how it might affect the broader ecosystem. The introduction of a genetically modified organism into the human microbiome could have ecological consequences such as disrupting the natural balance of skin bacteria or altering interactions within microbial communities.

Environmental Impact

Our engineered C. acnes strain expresses a protein intended to specifically target scabies mites. Importantly, this strain has biological characteristics that limit its ability to spread throughout the environment. As a slow-growing, facultatively anaerobic, non-spore-forming bacterium, C. acnes is well suited to the human body but does not naturally thrive outside of these specific niches. Its dependence on human-associated environments reduces the risk of accidental release into the wider environment, mitigating concerns regarding its impact on ecosystems beyond the skin.
However, the ecosystem associated with scabies could still be affected by the widespread use of this lotion. Scabies mites play a role in trophic networks and serve as prey for other organisms. A large-scale reduction in mite populations could disrupt these ecological interactions, potentially leading to unintended consequences for the local biodiversity. Understanding these potential impacts will require further research into the ecological role of scabies mites and how their removal could affect the food web in the affected areas.

Ethical Responsability

As scientists, we have the ethical responsibility to consider both the benefits and risks of our work. While our engineered lotion has the potential to improve public health by providing effective treatment for scabies, we must also be vigilant in ensuring that it does not cause unintended harm to individuals, communities, or the environment. This involves a commitment to transparency in our research, collaboration with regulatory bodies, and the ongoing monitoring of the product's impact once it is in use.
In conclusion, although our project has the potential to offer significant public health benefits, it also requires careful ethical consideration at every stage of development. From regulatory compliance and safety testing to understanding the potential risks and environmental impacts, we are dedicated to ensuring that our work is both scientifically sound and socially responsible.

BIBLIOGRAPHY

[1] Smith JA, Davis KL. Best practices for handling non-pathogenic microorganisms in the laboratory. Appl Microbiol Biotechnol. 2019;103(4):1625-34.
[2] McLellan PW, Lee SB. Safety practices in the microbiological laboratory: A review. J Lab Saf. 2018;10(3):112-20.
[3] Green RL, Walker AH. Decontamination and sterilization protocols for laboratory safety. Lab Saf Sci. 2021;17(1):45-58.
[4] Nelson MC, Thomas LR. Comparison of ethidium bromide and SYBR Safe for DNA gel electrophoresis: A safety perspective. Anal Biochem. 2020;606:113806.