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Safety

1.) The largest threat to researchers, the public, and the environment is potential contamination by the E. Coli bacteria. If this project were to be conducted on a large scale, large quantities of the bacteria would be necessary and would therefore increase risk. Both humans and animals could be harmed by the bacteria. Fortunately, if careful measures are taken to limit exposure to the bacteria and to sanitize areas that come into contact with the bacteria, this problem can be managed. The team has not received formal training but has taken care to disinfect materials that make contact with the bacteria. Though E. Coli used in the project can be harmful, it is something that is easy to obtain, so the risk of someone with malicious intent using the bacteria would not be significantly raised. The best way in which to avoid that kind of scenario is to limit access and exposure to the bacteria, especially when there are large amounts of it.

3.) Our school does not have a biosafety committee so we just considered the biosafety rules and regulations in place in the United States. [http://www.cdc.gov/biosafety/publications/bmbl5/bmbl.pdf The CDC Biosafety Manual] contains all these guidelines. We determined that E. Coli would be placed in BSL group 1, for the bacteria is abundant in nature and most strains, including the ones we are using, so it will not cause harm to individuals or the community. The NIH guidelines for recombinent DNA and genetically altered organisms state that the most important thing to consider is that "It is particularly important to address the possibility that the genetic modification could increase an agent’s pathogenicity or affect its susceptibility to antibiotics or other effective treatments" (CDC Manual). We considered this question and plan to, in the future, test how different acidity affects our bacteria as a way to control for this. For example, our bacteria might not function well at a certain acidity because it has a proton pumping gene, so this may be a good way to control for the problem of it becoming potentially widespread and out of control. Additionally, we adhered to standard biosafety techniques such as using primary barriers and personal barriers so as not to spread the bacteria. At this point in our research, the bacteria, even if it got out, would not be particularly virulent because the only effect is has on the environment is a color change. We decided, therefore, that basic safety measures are adequate to contain our work.

Lab Safety

While working with the E. Coli bacteria, our team took careful measures to avoid contamination. Any used pipette tips were soaked in bleach in order to kill the bacteria. Additionally, team members made sure to wash their hands after participating in the experiment. We made sure to tightly close and safely store any containers holding either bacteria or DNA samples. We monitored and took great measures to contain the unused bacteria and DNA samples as well.

How Our Bacteria Promotes Safety

There are no current technologies that assist growing plants that require a low pH. Farmers grow these particular plants, such as tomatoes and blueberries, in an artificially acidic environment. Adding acids to the soil can be detrimental to the environment in many aspects. The primary concern is what the consequences would be if the chemicals leaked out of the system and into surrounding areas. It could contaminate other plants that need a high pH to grow, thus killing crops that supply entire towns with food. There are also uncontrollable factors, such as acid rain or water runoff that can change the pH. Because of this, this method requires a lot of manpower to constantly maintain the correct pH for the plants to grow in.

Our solution is better than directly adding chemicals to the environment because it takes all of these issues into consideration. Using a bacteria that would be able to change the pH on its own is clearly the best method to growing plants in acidic conditions. Because the bacteria releases protons on the basis of the surrounding pH, we would not need to worry about uncontrollable environmental factors changing the pH because the bacteria does not take into consideration what is changing the pH, simply the quantitative data. We would not need to maintain the soil so regularly because bacteria reproduces in the soil by natural causes.

Right now, we were only capable of creating a pH sensor bacteria that changes color upon the detection of a pH of 5.5-8.0 (the optimal pH range for the growth of the majority of produce), which will offer a new method for farmers with crops that cover small to large acreage to test the pH of the area. This method is a more efficient method since the local and “representative” tests may not apply to the entire area where crops are being grown. By spraying the bacteria from a plane over a large field, the farmers will get a visual representation of which areas are at a proper pH and which are not. Not only will this bacteria be a more efficient method of pH testing for farmers, but it will be an excellent indicator of the areas affected by runoff since the color expressed would not be present in the area.

Nonvirulent and nonpathogenic E. Coli exists in nature in both commensal and symbiotic relationships with the environment around them. Our nonvirulent bacteria are very simple organisms that almost certainly not aversely affect the environment since they only have the sensor and the reporter genes along with the other natural E. Coli genes. The protein that they express is harmless and can be washed off of the plants and the area by water from rainfall and other sources.

Safety in the Field