Team:USC/Safety

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						<p style="text-align:center;"><span style="text-decoration:underline;color:#000000;"><strong>Identifying Safety Issues</strong></span></p>
<p style="text-align:left;"><span style="color:#000000;">Working in a research laboratory can be fun but also dangerous if safety for the student, society, and environment are not properly addressed. Therefore, safety issues of these three important entities will be examined under the scope of risk assessment. To assess risk for a biological material, many factors have to be considered, which include but are not limited to: the microbe&#8217;s pathogenicity and ability to survive outside of the lab, public awareness of the biological agent, availability of a prophylaxis, environmental impact, and ability to monitor biological material that are potentially dangerous.</span></p>
<p style="text-align:left;"><span style="color:#000000;"><strong><a href="http://uscigem2012.files.wordpress.com/2012/09/dsc_0003.jpg"><span style="color:#000000;"><img class="alignright" title="DSC_0003" src="http://uscigem2012.files.wordpress.com/2012/09/dsc_0003.jpg?w=366&h=243" alt="" width="366" height="243" /></span></a></strong></span></p>
<p style="text-align:left;"><span style="color:#000000;">We assess risk for the biological material involved in our project and its potential impact to the researchers, public society, and environment by taking into account two components of risk: probability and hazard. With probability, we ask ourselves:</span></p>
<ul style="text-align:left;">
<li><span style="color:#000000;">What are the likely and unlikely (including worst-case) scenarios of conducting research in our project?</span></li>
<li><span style="color:#000000;">Is there a potential risk that may compromise in any way researchers, society, or the environment? How likely that risk comes to fruition?</span></li>
</ul>
<p style="text-align:left;"><span style="color:#000000;">Questions to consider about hazards associated with biological material risk are:</span></p>
<ul style="text-align:left;">
<li><span style="color:#000000;">If the project works, what may be the potential hazards to researchers, society, and the environment?</span></li>
<li><span style="color:#000000;">By what mechanism may the engineered organism of the project affect researchers, society, and the environment?</span></li>
<li><span style="color:#000000;">If the engineered is used outside the lab, how many natural mutations to the the organism&#8217;s DNA affect and alter the functionality and concomitant risks to researchers, society, and the environment?</span></li>
</ul>
<p style="text-align:left;"><span style="color:#000000;">We assess biological safety risks with these questions in mind in an effort to make our project acceptably safe according to the standards set by organizations, such as the World Health Organization (WHO), the Convention on Biological Diversity, the National Institute of Health (NIH), American Biological Safety Association, Center for Disease Control, among other risk management groups, and conferences with an emphasis in synthetic biology and safety such as the SB5.0 conference. In addition, complying with these safety standards will make the project and its future directions more approachable for acceptance in public society.</span></p>
<p style="text-align:left;"><span style="color:#000000;"><strong><span style="text-decoration:underline;">Researcher Safety</span></strong></span></p>
<p><span style="color:#000000;">Biological Safety Provisions</span></p>
<p><span style="color:#000000;">At USC, the Department of Career and Protective Services (<a href="http://capsnet.usc.edu/LabSafety/biosafety/"><span style="color:#000000;">http://capsnet.usc.edu/LabSafety/biosafety/</span></a>) oversees that all research being done here, especially in biology, is safe. Initially, our team completed a laboratory safety class with the department before beginning work in the laboratory.</span></p>
<p><span style="color:#000000;">Our training included the following:</span></p>
<ul>
<li><span style="color:#000000;">Shared testimonials of accidents that have occurred in a laboratory and how we reacted to those accidents.</span></li>
<li><span style="color:#000000;">Participated in an activity involving how to use a MSDS.</span></li>
<li><span style="color:#000000;">Reviewed the meaning of common signs in the laboratories.</span></li>
<li><span style="color:#000000;">Reviewed what personal protective equipment we should be using.</span></li>
<li><span style="color:#000000;">Reviewed procedures during emergencies, including how to use an eye-wash station, shower, and how to dispose of a fire.</span></li>
<li><span style="color:#000000;">Learned where to dispose of all hazardous laboratory materials, biological waste, and chemical waste.</span></li>
<li><span style="color:#000000;">Received a certificate of our training.</span></li>
</ul>
<p><span style="color:#000000;">In addition to the training, the iGEM researchers also made sure to:</span></p>
<ul>
<li><span style="color:#000000;">Wear gloves, lab coats, and goggles in the laboratory at all times.</span></li>
<li><span style="color:#000000;">Take off all personal protective equipment when exiting the laboratory to work in the office.</span></li>
<li><span style="color:#000000;">Not eat in any part of the laboratory except in the office.</span></li>
<li><span style="color:#000000;">Clean up and organize the laboratory benches after each day of research.</span></li>
</ul>
<p style="text-align:left;"><span style="color:#000000;">Ensuring the safety of the USC iGEM lab and its researchers is critical to the success of the program. We have therefore taken initiative that each member understands and is aware of the following:</span></p>
<ul style="text-align:left;">
<li><span style="color:#000000;">Physiological and environmental risks associated in experimenting with a certain biological material.</span></li>
<li><span style="color:#000000;">Course of action to take, as standardized and/or recommended by risk management groups, if the biological material is inappropriately exposed to the researcher and/or the environment.</span></li>
<li><span style="color:#000000;">Knowledge of lab equipment to utilize and reduce the possibility of inappropriate biological material exposure and protect the researcher if such exposure occurs.</span></li>
<li><span style="color:#000000;">Knowledge of safety protocols and location of safety equipment in case of inappropriate exposure, including emergency situations.</span></li>
</ul>
<p style="text-align:left;"><span style="color:#000000;">The laboratory, where the USC iGEM team works, is certified for Biosafety Level 1. Researchers are required to wear personal protective equipment (PPE) which include goggles, gloves, and lab coats so that the eyes, hands, and body, respectively, are always protected from possible exposure to a biological material. Potentially harmful biological materials and chemicals are used daily in the lab. Below is a description of each one we encounter on a daily or limited basis as well as the measures taken to comply with USC&#8217;s stringent safety standards so that we maintain the highest quality of our safety:</span></p>
<ul style="text-align:left;">
<li><span style="color:#000000;">Ethidium Bromide (EtBr) is an aromatic chemical agent often used for staining of DNA during agarose gel electrophoresis. It binds with DNA and fluoresces with an orange color under UV light. EtBr may be a mutagen and carcinogen since it is believed to intercalate with double-stranded DNA, therefore inhibiting the DNA from undergoing important biological processes such as replication and transcription. Special care is definitely taken when handling EtBr. Proper PPE is used, and disposal of the chemical is handled by the researchers in accordance with EtBr&#8217;s Material and Safety Data Sheet (MSDS). Pipette tips exposed to EtBr are ejected to a chemical waste basket, which also include used agarose gels that contain a residual amount of EtBr. In addition, nitrile gloves are disposed of after pipetting EtBr to the agarose gel solution to prevent secondary exposure of WtBr.</span></li>
<li><span style="color:#000000;">Ultraviolet (UV) light is used for imaging of stained DNA in agarose gel electrophoresis. However, unprotected and direct exposure of UV light is known to be harmful to the skin and eyes and may lead to skin cancer and eye-related problems. UV light damages DNA by multiple mechanisms. One involves the binding of two adjacent thymine nucleotides in the DNA sequence to make a thymine dimer, which results in distortion and reduced functionality of the DNA. When imaging DNA from a gel, researchers ascertain that the UV light is turned off and only turned on when the gel has been properly placed inside the imaging machine and its door closed so UV light exposure is shielded from the researcher. Similar steps are taken when taking the gel out. In gel extraction, a protective shield is worn protecting the face from UV light exposure.</span></li>
<li><span style="color:#000000;">Descendants of two <em>E. coli</em> isolates, DH5a and MG1655, have been used as tools and model organisms in our project. These strains are known to be non-pathogenic and are extremely unlikely to survive outside of the lab or in human hosts, despite some conferring a variety of basic antibiotic resistances (e.g. ampicillin, kanomycin, chloramphenicol, and streptomycin). Nevertheless, prevention of contact from the bacteria is done by requiring the researchers to wear proper PPE and decontaminating all laboratory materials exposed to the bacteria with 10% bleach before disposal.</span></li>
</ul>
<p style="text-align:left;"><span style="color:#000000;"><a href="http://uscigem2012.files.wordpress.com/2012/08/dsc_0007.jpg"><span style="color:#000000;"><img class="alignleft size-thumbnail wp-image-81" title="DSC_0007" src="http://uscigem2012.files.wordpress.com/2012/08/dsc_0007.jpg?w=385&h=255" alt="" width="385" height="255" /></span></a></span></p>
<p style="text-align:left;"><span style="color:#000000;">To ensure the researchers of USC iGEM are acceptably safe from physical harm by biological material or chemical exposure, each team member attended a Safety Orientation Course conducted by USC&#8217;s Career and Protective Services (CAPS). In addition, the protocols used in our project have been approved by the Institutional Biosafety Committee.</span></p>
<p style="text-align:left;"><span style="text-decoration:underline;color:#000000;"><strong>Public Safety</strong></span></p>
<p style="text-align:left;"><span style="color:#000000;">Although the <em>E. coli</em> strains we are using are non-pathogenic, we remain aware that our project can potentially be a concern for public safety. By addressing this issue early, we lower the risk of fear and mistrust that can be met by the scrutiny of the general public when introducing a new biotechnological tool to the marketplace and diminish the uncalculated possibility of causing severe physical and non-physical harm. Our approach to public safety can be categorized as follows:</span></p>
<ul style="text-align:left;">
<li><span style="color:#000000;">How may our immediate surroundings, which include the university campus and its neighboring communities, be affected by our project?</span></li>
<li><span style="color:#000000;">How may Greater Los Angeles as well as the general public be affected by our project?</span></li>
</ul>
<p style="text-align:left;"><span style="color:#000000;">The USC iGEM lab is located within the USC campus, one that invites more than 30,000 students daily. In addition, the campus is located merely miles from Downtown Los Angeles and other neighboring cities where millions of people reside. The risk for infection from the bacteria we use is extremely low since we use strains that are engineered for laboratory experiments. These strains would be unable to survive in the outside environment. However, we still do the following to minimize this risk as much as we can:</span></p>
<ul style="text-align:left;">
<li><span style="color:#000000;">Before entering and after exiting from the lab, hands are washed with hot water and soap for at least 30 seconds.</span></li>
<li><span style="color:#000000;">Gloves are always removed upon leaving the lab.</span></li>
<li><span style="color:#000000;">No food or drinks are brought inside the lab.</span></li>
</ul>
<p style="text-align:left;"><span style="color:#000000;">These simple procedures are implemented to ensure that the public in our immediate surroundings is respected with our safety concerns for them.</span></p>
<p style="text-align:left;"><span style="color:#000000;"><strong><span style="text-decoration:underline;">Environmental Safety</strong></span></strong></p>
<p style="text-align:left;"><span style="color:#000000;">The safety of the surrounding environment should not be compromised. The <em>E. coli</em> strain is non-pathogenic and equipment and reagents are disinfected with the proper chemicals when cleaning materials to avoid spreading and contamination.</span></p>
<p style="text-align:left;"><span style="color:#000000;"><strong><span style="text-decoration:underline;">BioBrick Safety Issues</span></strong></span></p>
<p style="text-align:left;"><span style="color:#000000;">None of the new BioBrick parts that we made raise any safety issues. We designed BioBricks for flagella assembly and chemotaxis control, none of which are pathogenic.</span></p>
<p style="text-align:left;"><span style="color:#000000;"><strong><span style="text-decoration:underline;">Thoughts on How to Improve Safety in iGEM</span></strong></span></p>
<p style="text-align:left;"><span style="color:#000000;">iGEM teams could test antibiotic sensitivity in all bacteria strains periodically, making sure that these strains will not survive an antibiotic that we expect the strains to be sensitive to. Parts, devices, and systems could be made even safer by having iGEM require mandatory laboratory safety check-ups during the summer as well as submit documentation of the safe practices employed.</span></p>
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