Safety
When working in any laboratory, even if it is only your school`s chemistry lab, safety should undoubtedly be your main concern. Work safety includes everything from proper equipment such as protective gloves, glasses, lab coat etc., to adequate preparations and proper disposal of chemicals and organisms.
Our experience with lab safety
Prior to this project we had already worked in a laboratory, either at school, in a research facility or in some other way. This way we had acquired a lot of experience in this field and were fully aware the importance of lab safety. It is a responsibility you have towards yourself and others working with you.
That is why before even stepping into a laboratory we had a mandatory work and laboratory safety lecture at the National Institute of Chemistry. We revised and upgraded our previous knowledge on this topic and after passing the exam we were fully prepared and educated on the topic of safe laboratory work. Prof. dr. Marko Dolinar, a member of the Slovene Scientific committee for work with GMOs in contained use, gave us a lecture on genetically modified organism and concerns regarding their use and safety. Additionally, we also watched two educational videos, which were provided to us by iGEM and gave us further information on how to work with living microorganism and how to handle GMOs.
Chemicals
We mostly dealt with chemicals which are in general safe, for example agarose and peptone, but in case of chemicals such as butyric acid, hydrochloric acid, sodium hydroxide and ethidium bromide, special caution was required. When handling with ethidium bromide, which is considered mutagen, carcinogen and teratogen, we were especially careful. We consistently used the stronger (purple) nitrile gloves and were cautious not to touch unnecessary objects when using them to prevent contamination of surfaces and equipment with ethidium bromide and maintain the required level of safety. In any case, ethidium bromide can only be used in the so-called 'electrophoresis lab'. No equipement or consumables used for work with ethidium bromide is allowed to leave this lab (except for waste sealed in a bag).
Organisms
We worked in a Level 1 laboratory, because all the organism (Escherichia coli and Clostridium acetobutylicum) and parts we were using fall into this biosafety category. Both bacteria are safe to use since they pose a minimal potential threat to people in the laboratory and do not consistently cause disease in healthy adults. Nevertheless, we were precautious and used proper disinfectant to clean the laminar flow cabinet and other working areas as some of the E. coli we were using during our research was of course genetically modified. Working in a dedicated molecular biology lab our working space is separate from the rest of the facility. Our lab is a part of a department which again is separated from the rest of the building by a door that locks automatically, so that the access to the department is controlled. Work was preformed on an open lab table. Personal protective gear (lab coat and gloves) was worn at all times and was supplemented with eye and ear protection as needed (for example for work with UV light or sonication). All liquid waste was autoclaved before being discharged. Solid waste was collected separately and disposed by inicineration by an independent contractor. A sink for hand washing and 70% ethanol was available for sterilisation at all times if needed.
Butanol
Butanol as our main product, when produced in adequate quantities, would be used in standard Otto gasoline engines as a propellant. Since this engine is one of two most common engines, the other one being the diesel engine, we had to consider several aspects of butanol`s broad use, including safety and possible impact it would have on the environment.
Butanol is according to the CDC harmful to health. It causes damage if inhaled, ingested or if it comes in contact with skin or eyes. Inhalation of vapours may cause headache, nausea, drowsiness and dizziness and irritation of the respiratory tract. Ingestion may result in vomiting, nausea, diarrhoea, abdominal pain, dizziness and drowsiness. Exposure to the eyes and skin causes irritation, redness and pain.
1-Butanol has low water solubility, is biodegradable and therefore not expected to significantly accumulate in nature. In open spaces, at normal temperature, 1-butanol decomposes due to photolysis. It is a combustible and flammable liquid, sensible to static discharges. Flash point of butanol is 29°C so spontaneous ignition is possible on hot days.
But compared to gasoline butanol is much safer to use. Gasoline is more hazardous to health as butanol, because in addition to skin, eye and respiratory irritation, dizziness and drowsiness, it is also mutagenic, carcinogenic and toxic to reproduction, causes damage to organs (liver, kidneys, bladder, blood, bone marrow, nervous system) through prolonged or repeated exposure and may be fatal if swallowed or inhaled. Gasoline`s flash point is -43°C and as such significantly lower than the flash point of butanol. In addition gasoline poses a greater threat to the environment than butanol, because it is poisonous to aquatic life. Gasoline safety sheet
To conclude, common use of butanol as a propellant may require certain safety precautions during production, transportation and storage. Possible dangers of its use could be eliminated with the same preventive measures that are already implemented for other flammable liquid car fuels.
Butyric acid
Butyric acid is a medium-strong carboxylic acid. Because it is the first and most important substrate of our engineered metabolic path, its dangers should be taken into consideration before any large-scale production of butanol is established. Butyric acid is corrosive and can cause pain, redness and severe deep burns on skin and loss of vision. If ingested, it can cause abdominal pain, shock or collapse. Inhalation may result in sore throat, cough, burning sensation, shortness of breath and laboured breathing. Butyric acid safety sheet It is also combustible and forms explosive vapours above 72°C. With proper action these dangers should be eliminated.