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Team:Uppsala/Safety
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| - | document.getElementById("tab2").innerHTML = '<p> | + | document.getElementById("tab2").innerHTML = '<h2>LAB SAFETY</h2><p>When working in any laboratory environment, safety must always be addressed. We held safety rounds in the lab and organized a safety-day for the team to focus on discussing safety questions. </p><h2>Disinfection and sterilization</h2><p>It is important to disinfect and sterilize to ensure both the health of the labcrew as well as for the environment. Working with class 1 bacteria means that the potential harm to the labcrew is limited, but there is a pressing issue about the spread of antibiotic-resistance from the standard plasmids used in iGEM. In our lab we use several precautions to make sure that no bacteria are spread outside the lab.<br><br>All liquid bacterial cultures are gathered in glass containers. This also includes LB which has been in contact with bacteria, such as the supernatant of centrifuged cultures. The liquid is then treated with “Jodopax”, an iodine rich substance that is used in lower concentrations as a wound disinfector. In high concentrations Jodopax could cause rashes or smaller allergic reactions. To limit the exposure of Jodopax we have set it up in “spray bottles” so it could be inserted directly into the glass container. This is also done close to the sink so if skin is exposed to Jodopax it can easily be rinsed off. After at least one hour of treatment the solution is disposed in the sink. The glass container is then “flushed out” and then cleaned as normal glassware. All glass pipettes are put into buckets with “Virkon” solution to ensure sterilization. They are left in the buckets for at least one day. <br><br>Since our class one bacteria is not dangerous upon skin contact we have put our focus towards not letting the bacteria enter the body through e.g. the mouth. Therefore we do not use gloves. If you use gloves, you will probably not notice when you get bacteria on your gloves, later on you will spread the bacteria in the lab by touching things with your infected glove and you might even touch yourself in the eye or mouth. Instead we make sure to have 70% ethanol at all benches in the lab and disinfect frequently. As soon as we feel any drop or after working with bacteria we disinfect with ethanol. This way the bacteria is killed off straight away and its risk of spreading both in the environment and in us is reduced. <br><br>All one time use materials such as pipette tips, tubes and plastic loops are disposed in special hazardous waste containers. Swedish law states that the waste must be packaged such that it can not be re-opened and re-closed without notice. No material should be able to escape the box. To ensure this our bins consists of thick cardboard boxes with two thick plastic bags inside. In the inner bag an adsorption medium is placed to ensure that no liquid can escape. The boxes are changed each day and are then stored a maximum of five days in a cold room before transport or movement to a freezer storage for contagious waste. The waste is then transported via ADR certified trucks to local wasteincinerators “Vattenfall Värme AB” in Uppsala or “SVA:s förbränningsanläggning” in Ultuna. <br><br>To make sure that our working solutions of medium and plates are sterile we use autoclavation. Since autoclavation is a machine that builds up pressure and temperature it can be dangerous if handled incorrectly. To limit mistakes four persons from our team, that had previous experience from last years team, were trained to use the autoclave. They were the only ones that were allowed to use the autoclave from there on. </p><h2>Hazardous chemicals</h2><p>It is not only bacteria that pose a threat to the lab workers, there are also several chemicals that especially in a Class 1 lab are often more dangerous than the bacteria itself. In our lab we are working with EtBr, Acrylamide, Antibiotics and acids & bases. We have taken precautions when working with all these.<br><br><u>Etbr:</u><br>EtBr is a substance used to stain nucleotides , this means that it might hurt our own DNA or RNA if it can penetrate the cell membrane. At first we used gelred as a substitute to lower the risk. Gelred is a EtBr molecule that has a carbon chain attached to it to limit its possibilities to enter through the membrane. However the gelred ran out and the personnel responsible for ordering reagents were on vacation, and thus we had to use EtBr. To limit exposure of EtBr we decided to stain via shaking gels that have finished running in water with EtBr. All work with EtBr was done with nitrile gloves, lab coats and goggles in a fume hood to limit exposure and spread. From the part when the gel had finished running we always worked in pairs. One of them, that touched EtBr, was not allowed to touch anything outside the cabinet, the other person helped by being able to touch things. Before disposal of the shaking bath with EtBr we neutralized it with “teabags” that degrade the EtBr for more than 24 hours.<br><br><u>Acrylamide:</u><br>Acrylamide is a cancerogenic substance that is used for the gel in SDS page. To protect us from acrylamide, we used nitrile gloves, lab coats and safety glasses. Only the person who worked with SDS page was allowed at the bench with acrylamide. Similarly the work was done in pairs as with the EtBr. All materials that have been in contact with the Acrylamid was disposed in the contagious waste. <br><br><u>Antibiotics:</u><br>Antibiotics are at risk of increasing the resistance of antibiotics if released into the environment. This might affect health care leading to decreased effective treatments for everyday diseases. Swedish law classify it as a “Drug” waste. Antibiotics can also be somewhat toxic in high concentrations, so gloves, labcoats and glasses were worn at all times when creating stock solutions of antibiotics. <br><br>Concentrated antibiotics together with one-time use material was sorted into contagious waste boxes as mentioned above while small liquid cultures can be treated with “Jodopax” to destruct the chemical.<br><br><u>Acids & bases:</u><br>All members of the team had taken at least two courses in chemistry, which of one was “Organic Chemistry”, a course known to handle concentrated strong acids and bases. In our lab we use NaOH pellets to get correct pH for our TBE and Boric Acid. They are handled with care and lab coats, gloves and goggles and always in a fume hood. The fume hood is very important when working with boric acid since it can be poisonous through inhalation of gas.</p><h2>Class 2 organisms</h2><p>We were able to work with the real <i>Y.enterocolitica</i> at “Livsmedelsverket”, ‘Food Administration of Sweden’, in their class 2 lab. Since they work with <i>Y.enterocolitica</i> every day, well thought out routines were held. A difference from WHO recommendations are the use of gloves. At Livsmedelsverket we almost never wore gloves. This is to ensure that you notice when you get bacteria on yourself so you disinfect straight away. No fire disinfection was used to limit the creation of aerosols, instead plastic one time use material was used. <br><br>To access the lab several restricted access area doors had to be passed, and the door was marked with a biohazard symbol and “Class 2 lab” logo. The lab is authorized by Swedish lab security agency. </p><h2>Lab security</h2><p>Most things in a laboratory can be used for malicious purposes, especially chemicals and basic synthetic biology. It is therefore important to address the issue of lab security. The main concern for our project would be the genes we had synthesised and in what ways they could be used in the wrong hands. We dropped the idea of collagen early on when discovering it could be harmful. Other genes such as USP45, the anchor protein and antibiotic resistance could probably be used with synthetic biology combined with other genes to create something dangerous. We also store chemicals as stated above that are potentially dangerous. They are stored in alphabetical order, which makes it easier for someone that might want to use them maliciously. However our lab is a restricted access area and only us, and the lab officers have access to the area. Further more, whatever we might have in the lab there are probably a lot of easier ways for these kind of individuals to get their hands on it, including the real <i>Yersinia Enterocolitica</i> at livsmedelsverket.</p><h2>Sources and inspiration</h2><p>Our methods in safety and security are mostly based upon oral information from our supervisors and lab officers. Uppsala University has a well described document for lab safety which we follow, called “Riskhantering vid Uppsala Universitet” loosely translated to ‘Handling of hazards at Uppsala University’. In that document references to Swedish laws and regulations are also made. To make sure that we were also following international guidelines we decided to read the “WHO biosafety manual”, this gave us inspiration and thoughts about the safety issues with designing a system that should be addressed.</p>'; |
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Revision as of 13:21, 16 October 2014
Stephanie Herman
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