Safety

Project procedures for safety

Here at CAPS, we make safety our number one priority when we plan and execute our experiments. Situational awareness and communication is essential for any success. When using cyanobacteria or chemicals, we always work in sterilized environment. It’s important not to contaminate our sample with ourselves or surroundings and not to contaminate us with our sample! At all times, we wear our personal protection equipment (PPE) which consists of disposable gloves, lab coats, and eye protection. We follow a safety handling protocol suggested by one of our mentors to safely and efficiently handle cyanobacteria. A mentor or qualified adult is always present to be an extra set of eyes. We know what we are going to do before we set foot in the lab.
Researcher Safety
Everyone on the team obtained laboratory safety training through the Machinery of Life/Foundations of Molecular Medicine and Bioengineering CAPS Class and had experience with bacterial culture (in E.coli) and genetic transformation (inserting pGLO into E. coli) prior to beginning the project.
During the work, our team took the following safety precautions

1. Lab coats, gloves and goggles were worn at all times.
2. We were supervised in both the CAPS lab and at Dr. Fenton’s lab down KU Med.
3. All equipment (both disposable and autoclaved) was sterile.
4. The bench and other work surfaces were sterilized with water and bleach before and after lab work.
5. Because cyanobacteria can pose a risk if ingested, there was no food or drink in the lab.
6. There were no lit burners or other fire during lab work.

Organism and Part Safety
Because our project utilizes the mixotrophic cyanobacteria Synechocytis PCC6803 and E. coli (two BSL-1 organisms) safety risks may initially appear to be minimal, almost nonexistent. Neither of our organisms are pathogenic. However, upon closer examination, our project contains elements that call for considerable thought towards safety.
First, we recognize that cyanobacteria are autotrophs and require little other than water and light to survive. Therefore, if we did not practice sterile technique, it would be possible for cyanobacteria to be dripped or spilled and survive.
Cyanobacteria produce enzyme-inhibiting toxin that can affect the nervous system, immune system, and liver. These toxins have killed waterfowl and marine mammals that drank from algal bloom-contaminated water. However, the worst cyantoxins, cyclic petides, alkaloids, and ketides are not produced by our chassis, Synechocytis 6803.
Alkanes are biologically inactive and impose a small but present risk. Many of the alkanes that are gases or liquids at room temperature (most notably, ethane, methane, propane and butane) are flammable. These gaseous alkanes are not toxic and would only pose a danger if we breathed them in such high concentrations (which would have to be twelve to sixteen percent) that we suffered from oxygen deficiency.
Environmental Safety
However, the main risk deals not with toxins themselves but with the dangerous of cyanobacteria to the environment and the risks our altered Synechocytis PCC6803 might pose to the ecosystem. Besides the inherent risk of introducing any genetically modified organism into the world outside the laboratory, we recognize that our Synechocytis 6803, like any cyanobacteria, could become the culprit in an algal bloom. Per iGem safety suggestions, we consider the worst-case scenario.
Harmful algal blooms (HABs) are essentially proliferations (often as dense as millions of bacteria per milliliter) of cyanobacteria in salt or freshwater. The water may appear entirely green and become so turgid that animals that hunt by sight cannot see more than a centimeter or two. The cyanobacteria may consume all the oxygen in the water, leaving none for other organisms, and when they die, they sink to the bottom of the body of water, and can suffocate organisms there. Though some algal blooms can be natural (for example, those that are believed to be related to El Nino) many are the direct result of human eunutrition. Runoff from fertilizers puts high levels of phosphates and nitrogen into water, creating an ideal environment for cyanobacteria growth.
If our alkane-producing cyanobacteria were to be the culprit in an algal bloom, the effects might be worse than if an unaltered cyanobacteria proliferated in the bloom. Because alkanes are a major component of gasoline and diesel, the effects might be similar to an oil spill. Furthermore, certain bacteria and archea are able to metabolize alkanes, possibly leading to an out-of-control proliferation of these organisms.
Ethical Safety
We recognize that our rabbit pyruvate kinase gene was isolated by Dr. Fenton and that we only get to use it per his permission. Accordingly, we have kept him informed on our work with it and the possibility of this gene becoming readily available in the Standard Registry of Biological Parts.