EH&S regulates safety for all research at UCSC. These standards are used to advise faculty and individuals regarding their responsibilities concerning safety, health, and environmental issues, and recommend corrective actions and safety programs. EH&S helps provide all labs with personal protective equipment (PPE), which includes lab coats, goggles, and gloves.
Safety
Safety is our top priority, both in how we handled materials and protocols in the wet lab. We also took time and consideration as to how the final product interacts with consumers and affects the environment. Our team was dedicated to maintaining high safety standards, and to achieve this, each team member was required to undergo extensive safety training before beginning work in the lab.
Environmental Health & Safety (EH&S) Standards
UCSC iGEM Standards
Before beginning the wet lab component, all team members and interns were trained in the necessary safety protocols by our Principal Investigator, David Bernick. If there was wet lab work to be done, at least two people were required to be in the lab at all times. We were expected to keep each other accountable and inform the PI in case of an emergency. Food and beverages, including water, were not permitted within the lab space to ensure the sterility of our lab equipment, as well as the safety of individuals in the lab. Any machines used were turned off and unplugged after use to avoid the risk of fire. In the lab, we often worked with dangerous chemicals. Before following a protocol, we made sure that we were familiar with the reagents we planned to use and asked our PI in advance if we were unfamiliar with them. All pipette tips, plates, gloves, and plastic tubes that have come into contact with biohazardous material were properly disposed of in our lab’s biohazardous waste bin. All sharps, such as broken glassware, were disposed of properly in our lab’s sharps waste container. Our lab was also equipped with a working fire extinguisher, eye wash station, and safety shower in case of emergencies.
Malice Analysis Standards
We also worked with the Malice Analysis team through the Engineering Biology Research Consortium, which advises researchers on how to incorporate biosafety considerations and practices into their research design.
PPE
When in the lab, proper PPE is enforced, which consists of a lab coat, gloves, safety goggles, and unexposed skin through wearing long pants and closed-toed shoes.
Aflatoxin B1 & Mycotoxin Safety
Aflatoxin B1, like many other mycotoxins, is extremely toxic and carcinogenic. Rather than exposing our team to such harmful substances, we used progesterone as an analog in experimental procedures due to its similar structure and molecular weight to aflatoxin B1. Despite being a safer option, progesterone can be dangerous when ingested by individuals of both sexes, and our team has addressed this through training of the proper lab safety protocols.
Dangerous Chemical Safety
Dangerous or corrosive chemicals were used in various experiments. SYBRGreen and SYBRGold are intercalating agents that can cause DNA damage when ingested or in direct contact. Formamide is a carcinogen, teratogen, and is damaging to the blood. Chloroform and ethanol are both flammable, and in addition, chloroform is also toxic, carcinogenic, and can cause organ damage and CNS depression. The above hazards could be accidentally disposed of in the sink, posing an environmental risk, or ingested, which can cause skin and nasal irritation. Formamide specifically can become a risk if inhaled, which could occur if it is not used properly under a fume hood.
Potential Dual Usage Statement
While our aptamer product is intended to remove aflatoxin B1 from aqueous solutions and limit contamination, it can possibly be used with malicious intent as a toxin concentration and delivery vehicle in the wrong hands. Furthermore, naturally competent cells in the environment could take up plasmids if they are released, acquire antibiotic immunity, and concentrate mycotoxin from their surrounding environment intercellularly, which can impact the food chain. Validation of composting to safely dispose of aflatoxin, efficacy of the product in aqueous consumer foods, validation of lay public usage of this tool, and E&HS, FDA, and university approval are needed before releasing our product to the public.
Reagents
We used Escherichia coli bacteria, specifically strains DH5a, Top10, and BL21, specifically DH3. Most of our reagents are from IGSC-based companies such as IDT and Thermo Fisher.
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