Safety

Overview General Laboratory Safety
Laboratory and Equipment Waste Disposal
Personal Safety Protection Experimental Safety
Prevention of Protocols' hazards Lab Risk Scenario and Response Plans
Safety in Future Applications References

Overview

Safety is a top priority throughout our project. We proactively identified potential risks associated with both laboratory work and project application, developing preventive strategies to mitigate these challenges. All of our experiments are designed and conducted in accordance with iGEM safety guidelines, University of Macau’s biosafety regulations, and best practices in synthetic biology in order to prevent harm to ourselves and the environment. We also consider the potential risks in the experiment and future application of our project and find the appropriate response to solve and prevent these risks.

General Laboratory Safety


Laboratory and Equipment

We conducted our experiments (excluding the experiment of termites) in our iGEM lab, which is located in the building of the Faculty of Health Science, classified as BSL-2. Safety principle compliance remains our top priority in every phase of our experiment, and the equipment is regularly inspected by the university in order to maintain it in good condition. Our laboratory is equipped with the following safety facilities and emergency items (Figure 1-5):

  1. Chemical Fume Hood
  2. Emergency Button
  3. Oxygen Level Detectors and Lower Explosive Limit Detectors
  4. Fire Alarm System and Fire Fighting Equipment
  5. Emergency Eye Wash and Shower
  6. First Aid Box and Spill Kit
  7. Personal Protective Equipment (hand protection, eye protection, masks and respirators, etc.)

Fig. 1 First Aid Box and Oxygen Level Detector in our lab


Fig. 2 Spill Kit in our lab


Fig. 3 Fire Alarm system in our lab


Fig. 4 Fire Fighting Equipment in our lab


Fig. 5 Emergency Eye Wash and Shower in our lab


Waste Disposal

In order to secure a better environment for our work, the manner in which we dispose of waste stands out as a vital process.

  • All biohazard waste was disposed of in the biohazard bin located in the laboratory.
  • Biologically contaminated waste was mixed with chemical disinfectant, such as bleach, before pouring.
  • Sharp, needles, and syringes are disposed of in a sharp bin.
  • Broken glass were disposed of in a broken glass box.

Fig. 6 Regular Garbage bin and Biohazard Bin in our lab


Fig. 7 Sharp Bin in our lab

Personal Safety Protection

Before conducting the experiments in the lab, our team members have attended the safety tour and training provided by Univeristy of Macau, and need to pass the lab safety exam so as to ensure we understand the lab safety and survival in the lab. The safety training and exam contains the content of personal protective protocols, the identification of common hazards, and procedures for addressing incidents. Furthermore, we require that at least one instructor be present during all experimental activities to offer guidance and uphold continuous safety supervision for the full duration of the work.

We should obey the personal safety requirements when we work in the lab. First, we should wear the required personal protective equipment (PPE) such as a lab coat, safety goggles, and gloves, and always wash our hands thoroughly before leaving the lab. Second, we should have proper personal attire and appearance, like wearing closed-toe shoes and tying back long hair. Third, we never eat, drink, smoke, or store food/drinks in the lab, and keep the work area tidy and organized.

Experimental Safety

We have mentioned potential project risks, such as chemical hazards, in our Safety Form. We pay attention to safety throughout our experiment to avoid harm to human health and team members or colleagues in the laboratory.

Prevention of Protocols' hazards

As we have two parts of the experiment in our project, which are attracting termites and killing termites, we cultured the bacteria and used different chemicals that have a potential risk for people. Therefore, we should provide solutions for these hazards.

Part 1: Chemicals for Attracting Termites

This part involves the production of the attractants Hydroquinone (HQ) and its protector, Tea Polyphenol.

  1. Benzene

    2. Hazards:

    • Volatile liquid; inhalation of vapors is the primary exposure risk.
    • Chronic exposure can cause damage to bone marrow and significantly increase the risk of leukemia.

    Safety precautions:

    • Handle only inside a certified fume hood.
    • Use sealed, airtight containers for storage.
    • Wear appropriate PPE: nitrile gloves, lab coat, and safety glasses.
    • Monitor air quality if used frequently.

  2. Phenol

    3. Hazards:

    • Highly corrosive; causes severe chemical burns upon skin contact.
    • Vapors are irritating to the eyes and respiratory tract.

    Safety precautions:

    • Handle in a fume hood.
    • Wear chemical-resistant gloves (e.g., neoprene), a lab coat, and safety goggles or a face shield.
    • Have an emergency spill kit specifically for phenol readily available.

Part 2: Chemicals for Killing Termites

This part involves the production, purification, and analysis of the protein toxins (EcTI, Melittin, Ligand-Hecate). This includes lab techniques like SDS-PAGE, protein purification, and Western Blot.

  1. Acrylamide (for SDS-PAGE)

    2. Hazards:

    • Neurotoxins can be absorbed through the skin upon contact.
    • Repeated exposure can lead to nerve damage.

    Safety precautions:

    • Weigh solid powder entirely inside a fume hood to avoid inhaling dust.
    • Always wear double nitrile gloves and a lab coat when handling both powder and solution.
    • Use dedicated trays to contain spills when transporting bottles.

  2. Ethanol

    3. Hazards:

    • Highly flammable liquid and vapor.
    • Vapors can accumulate and ignite if exposed to an open flame, spark, or heat source, leading to fire or explosion.

    Safety precautions:

    • Use in a well-ventilated area away from ignition sources
    • Keep containers tightly sealed when not in use.
    • Have a fire extinguisher (Class B) and a fire blanket easily accessible.

  3. NaOH (Sodium Hydroxide)

    4. Hazards:

    • Strong base; highly corrosive.
    • Causes severe burns to the skin and eyes. Inhalation of dust or mists irritates the respiratory tract.

    Safety precautions:

    • Always wear splash-proof goggles, a lab coat, and gloves.
    • When diluting, always add pellets slowly to cold water (never water to solid) to avoid violent boiling and splashing.
    • Work in a fume hood when handling concentrated solutions or powders.

  4. SDS (Sodium Dodecyl Sulfate)

    5. Hazards:

    • Powder is an irritant; inhalation can cause respiratory irritation (coughing, shortness of breath).
    • Skin contact can cause dryness and irritation.

    Safety precautions:

    • Weigh powder in a fume hood to avoid creating airborne dust.
    • Wear a dust mask, gloves, and safety glasses when handling the powder.

  5. β-mercaptoethanol

    6. Hazards:

    • Has an extremely strong, unpleasant odor.
    • Toxic by inhalation and skin absorption. It can cause damage to the respiratory system and central nervous system.

    Safety precautions:

    • Handle exclusively in a fume hood.
    • Use sealed containers and ensure bottles are tightly closed immediately after use.
    • Wear double gloves, a lab coat, and safety glasses.

  6. Methanol

    7. Hazards:

    • Highly flammable liquid and vapor.
    • Toxic if ingested, inhaled, or absorbed through the skin; can cause blindness or organ damage.

    Safety precautions:

    • Handle in a well-ventilated area away from ignition sources.
    • Wear appropriate PPE: gloves, a lab coat, and safety glasses.
    • Always use a pipette aid.

  7. TRI reagent (for RNA extraction)

    8. Hazards:

    • Contains phenol and other corrosive components.
    • Causes severe skin burns and eye damage. Vapors are irritating.

    Safety precautions:

    • Handle all steps involving the reagent in a fume hood.
    • Wear nitrile gloves, a lab coat, and safety glasses.
    • Use tubes with secure caps to prevent leakage during shaking/centrifugation.

  8. IPTG (Isopropyl β-D-1-thiogalactopyranoside)

    9. Hazards:

    • Generally considered low toxicity.
    • Accidental ingestion may cause gastrointestinal discomfort.

    Safety precautions:

    • Standard laboratory hygiene: wear gloves and a lab coat.
    • Avoid creating aerosols.

  9. Imidazole

    10. Hazards:

    • Irritating to the eyes, skin, and respiratory tract.

    Safety precautions:

    • Wear standard PPE: gloves, a lab coat, and safety glasses.
    • Handle powders in a fume hood to avoid inhaling dust.

  10. Triton X – 100

    11. Hazards:

    • Causes skin and eye irritation.

    Safety precautions:

    • Wear standard PPE: gloves, lab coat, and safety glasses.
    • Avoid skin contact and aerosol formation.

  11. ECL reagents (Enhanced Chemiluminescence reagents)

    12. Hazards:

    • Components are toxic and may be carcinogenic.
    • Harmful by skin contact or inhalation.

    Safety precautions:

    • Handle with gloves in a well-ventilated area to minimize vapor exposure.
    • Change gloves immediately after handling to prevent contamination of other surfaces.

  12. EDTA (Ethylenediaminetetraacetic acid)

    13. Hazards:

    • It can chelate metal ions in the body if ingested in large quantities.
    • May cause mild skin irritation.

    Safety precautions:

    • Standard laboratory hygiene: wear gloves and a lab coat.
    • Avoid creating dust when weighing the powder.

  13. Potassium acetate

    14. Hazards:

    • Powder may irritate the respiratory tract and skin.

    Safety precautions:

    • Wear gloves, safety glasses, and a lab coat.
    • Handle powders in a fume hood or wear a dust mask to avoid inhalation.


Lab Risk Scenario and Response Plans

In any case of accidents, we should report to our supervisor and the Faculty Office as soon as possible.

Risk Scenario Immediate Response
Chemical Spill
  1. Evacuate the area immediately and alert everyone nearby.
  2. Ventilate the area by opening windows or turning on the fume hood.
  3. Contain the spill with absorbent pads or spill kits if safe to do so.
Fire
  1. Alert everyone and activate the fire alarm.
  2. Evacuate the area immediately and close all doors behind you.
  3. Use a Class B (flammable liquids) fire extinguisher only if the fire is very small and you are trained.
  4. Call emergency services from a safe location.
Skin or Eye Exposure to a Corrosive Chemical
  1. Flush affected area immediately with copious amounts of running water for at least 15–20 minutes.
  2. Remove any contaminated clothing while flushing.
  3. Seek immediate medical attention after flushing.
Cut or Laceration from Broken Glassware
  1. Apply direct pressure with a clean cloth to stop the bleeding.
  2. Wash the wound gently with soap and water.
  3. Seek medical attention for deep cuts or if bleeding does not stop.
  4. Dispose of broken glass in a designated sharps container.
Minor Burn
  1. Cool the burn immediately under cool running water for at least 10 minutes.
  2. Cover the burn with a sterile, non-adhesive dressing.
  3. Seek medical attention if the burn is large, deep, or blistered.

Safety in Future Applications

As synthetic biology-driven solutions of our project advance toward real-world application, ensuring safety across ecological, human health, and environmental dimensions becomes important. We consider safety for future applications, address potential risks and propose evidence-based mitigation strategies.

First Problem: Non-Target Organism Impact

The use of attractants such as hydroquinone and toxins like EcTI, melittin raises concerns about unintended effects on non-target species. For instance, HQ’s feeding- attractant properties may inadvertently lure beneficial insects or soil fauna, while broad-spectrum toxins could disrupt digestive systems of non-termite organisms.

Safety precautions:

  • Limit the scope of toxin action by structurally modifying EcTI to recognize only specific trypsin subtypes in the termite gut. Similarly, incorporate targeting peptides into melittin and hecate to ensure they act solely on protozoa unique to the termite gut.
  • Design the physical product (e.g., bait station with specific entry hole size and placement depth) to be accessible primarily to termites.
Second Problem: Environmental Release Risks

Engineered E. coli strains used to produce HQ and toxins pose genetic contamination risks if they escape. Furthermore, chemical residues like HQ and tea polyphenol complexes may persist and accumulate in soil or water.

Safety precautions:

  • Establish environmental monitoring methods for HQ and tea polyphenol residues and explore adding specific microbial degraders to the formulation to accelerate their breakdown, ensuring residue levels remain below ecological safety thresholds. For example, electrochemical sensor can be used for simple and sensitive determination of Hydroquinone in water samples. [1]
Third Problem: Human Health and Safety

Toxins such as EcTI and melittin could pose risks through skin contact, inhalation, or accidental ingestion during handling. Melittin, in particular, has the potential to cause allergic reactions in some individuals.

Safety precautions:

  • Integrate safety features into the product design, such as adding a warning bittering agent to solid baits to prevent accidental ingestion by children or animals, and using sealed injection devices for liquid products to minimize skin contact.
Fourth Problem: Ecosystem Chain Reactions

A sharp reduction in termite populations, which act as decomposers and a food source, could disrupt soil fertility, organic matter decomposition, and the food web for anteaters and certain birds. Altering the termite gut microbiome might also impact broader carbon and nutrient cycles.

Safety precautions:

  • Conduct dynamic ecological impact assessments in pilot areas, monitoring termite population density, soil microbial diversity, and populations of termite -predatory species. If signs of imbalance appear, adjust bait deployment or switch to milder, more specific control mechanisms. For instance, intelligent monitoring devices in termite control can be applied to conduct real-time monitoring of termite population density, soil microbial diversity, and populations of natural enemies. [2]
Fifth Problem: Technical Controllability and Supervision

Potential instability in genetic editing systems (e.g., mutations in CYP450-BM3) could lead to unexpected expression products. Improper product use, such as incorrect placement of bait stations or injection doses, could increase environmental and health risks.

Safety precautions:

  • Enhance the stability of genetic systems through methods like site-directed mutagenesis to improve enzyme stability and incorporate synthetic "safety switch" elements that allow precise control of gene expression via external inducers (e.g., IPTG).
  • Establish a full-chain supervision system, including detailed product usage specifications (e.g., burying solid bait boxes at least 30 cm deep to prevent wash- away) and regular safety audits in collaboration with environmental protection departments (e.g., soil microbe gene sequencing, water toxicity tests).

References

  1. [1] Karami-Kolmoti, P., Beitollahi, H., & Modiri, S. (2023). Electrochemical sensor for simple and sensitive determination of hydroquinone in water samples using modified glassy carbon electrode. Biomedicines, 11(7), 1869. https://doi.org/10.3390/biomedicines11071869

  2. [2] Nanxian Forestry Bureau. (2025, September 26). Nanxian Forestry Bureau implements precise termite control actions to build an ecological security barrier [Web page]. Hunan Forestry and Grassland Bureau. https://lyj.hunan.gov.cn/lyj/xxgk_71167/gzdt/xlkb/xsqxx/202509/t20250926_33815269.html

  3. [3] University of Macau, Office of Health, Safety and Environmental Affairs. (n.d.). HSEO Guidelines. https://hseo.um.edu.mo/hseo-guidelines/

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