Lab Safety: RAMP & Historical Hazards

• Laboratory safety is vital for minimizing workplace injuries and illnesses.
• It includes hazard recognition, risk assessment, risk minimization, and emergency preparedness.

RAMP Principles of Safety

• Recognize hazards.
• Assess risks of hazards.
• Minimize risks of hazards.
• Prepare for emergencies.

Historical Background on Laboratory Hazards

• A laboratory hazard is anything that can harm a vulnerable target.
• Several past incidents highlight the importance of laboratory safety.

Marburg Virus Outbreak (1967)

• In 1967, Marburg Laboratory workers in Germany experienced fever, diarrhea, vomiting, and internal bleeding.
• Seven workers died.
• The outbreak was caused by a previously unknown virus carried by monkeys from Uganda used for polio research.
• The virus was named after Marburg, the city of its discovery.

Chernobyl Disaster (1986)

• In 1986, a safety test at the Chernobyl nuclear power plant in Ukraine led to a catastrophic explosion and meltdown.
• It released massive amounts of radioactive material into the environment.
• The disaster resulted in immediate deaths, long-term health effects, and severe environmental contamination.

Karen Wetterhahn Mercury Poisoning (1997)

• In 1997, Professor Karen H Wetterhahn at Dartmouth College was exposed to dimethylmercury during an experiment.
• A few drops of the compound penetrated her latex gloves and reached her skin.
• She suffered severe mercury poisoning, leading to neurological damage and death within months.
• The incident showed that latex gloves are inadequate protection against penetrating toxic chemicals.

University of Minnesota Glass Vessel Explosion (2008)

• In 2008, a researcher at the University of Minnesota lost an eye when a glass vessel exploded under pressure.
• The accident was due to a lack of proper shielding and safety protocols for pressurized experiments.

University of California Electrical Arc Flash (2012)

• In 2012, a graduate student at the University of California suffered severe burns from an electrical arc flash.
• The incident occurred due to a lack of proper training and safety protocols for electrical work.

Gold King Mine Wastewater Spill (2015)

• In 2015, a cleanup operation at the Gold King Mine in the USA accidentally released 3 million gallons of toxic wastewater into a river.
• Poor planning and a lack of environmental safety protocols led to this environmental disaster.

Hazards in Cell Culture Laboratories

• Cell culture laboratories involve hazards from handling human or animal cells, tissues, and toxic or mutagenic reagents.
• Accidental punctures, spills, splashes, ingestion, and inhalation of infectious aerosols are common hazards.

Types of Hazards

Chemical Hazards

• Chemical hazards include toxic, corrosive, or irritating substances.
• Examples are medications, solutions, gases, vapors, aerosols, and particulate matter.
• Some chemical reactions generate heat, leading to thermal burns.
• Inhalation of toxic solvents and ingestion of chemicals are significant risks.
• Food and drinks should be stored away from chemical exposure.

Physical Hazards

• Physical hazards involve factors such as improper handling of equipment, extreme temperatures, noise, or vibration.
• Common injuries include cuts from glassware or sharp objects and burns from hot apparatus.
• Safe handling practices are essential to prevent injuries.

Electrical Hazards

• Electrical hazards arise from unsafe electrical systems, such as faulty wiring or equipment near liquids.
• Electrical fires can occur due to unsafe cords or plugs.
• Ground-fault circuit interrupters should be used to prevent electrical shocks.

Radiological Hazards

• Radiological hazards involve the uncontrolled release of radioactive materials.
• Proper shielding and handling protocols are necessary to prevent exposure.

Biological Hazards

• Biological hazards include bacteria, viruses, blood, tissues, and bodily fluids that can carry diseases or allergens.
• These hazards can be transmitted to laboratory workers.

Importance of Laboratory Safety

• Laboratory safety is crucial for reducing injuries and illnesses.
• Workers can protect their health and lives by minimizing or eliminating exposure to hazards.

Biosafety in Cell Culture Laboratory

• The primary goal is to minimize or eliminate exposure to harmful biological agents.
• Strict adherence to standard microbiological practices and techniques.
• Safety equipment includes primary barriers (e.g., biosafety cabinets) and personal protective equipment (PPE).
• Material Safety Data Sheets (MSDS) are also important for hazard communication.

Biosafety Cabinets

• Biosafety cabinets minimize exposure to hazardous materials.
• They provide an aseptic work area while containing infectious splashes or aerosols.
• Three classes of biosafety cabinets (Class I, II, and III) meet varying research and clinical needs.

Class I Biosafety Cabinets

• Protection for laboratory personnel and the environment.
• Do not protect cultures from contamination.
• Similar in design and airflow to chemical fume hoods.

Class II Biosafety Cabinets

• Designed for work involving BSL-1, 2, and 3 materials.
• Provide an aseptic environment for cell culture experiments.
• Protect personnel and the environment.
• Commonly used in cell culture laboratories.

Class III Biosafety Cabinets

• Gas-tight and provide the highest level of protection for personnel and the environment.
• Required for work involving known human pathogens and BSL-4 materials.
• Used in high-containment laboratories.

Air-Flow Characteristics of Cell Culture Hoods

• Biosafety cabinets protect the working environment from dust and airborne contaminants.
• They maintain a constant, unidirectional flow of HEPA-filtered air over the work area.
• HEPA filters can remove at least 99.97% of airborne particles as small as 0.3 microns.
• Airflow can be horizontal or vertical.

Personal Protective Equipment (PPE)

• PPE forms an immediate barrier between personnel and hazardous agents.
• It includes items such as gloves, laboratory coats, gowns, shoe covers, boots, respirators, face shields, safety glasses, and goggles.
• PPE is often used in combination with biosafety cabinets and other containment devices.

Material Safety Data Sheets (MSDS)

• An MSDS contains detailed information about the properties of a specific substance.
• Includes physical data, hazard information, and safety guidelines.

Biosafety Levels (BSL)

• Biosafety Levels (BSL) are biocontainment precautions to protect laboratory personnel, the environment, and the community from exposure to infectious agents.
• Levels range from BSL-1 (minimal risk) to BSL-4 (highest risk).

Biosafety Level 1 (BSL-1)

• Minimal risk to personnel and the environment.
• Appropriate for agents not known to cause disease in healthy humans (non-pathogenic, such as E. coli).
• Requires standard microbiological practices, such as handwashing.
• Work can be performed on open bench tops.
• Required safety equipment includes basic PPE, such as lab coats, gloves, and safety glasses.
• Commonly found in teaching laboratories for undergraduate students.

Biosafety Level 2 (BSL-2)

• Involves moderate risk, where agents pose a hazard to personnel but are unlikely to spread to the community.
• Examples include pathogens that cause mild to moderate disease, such as Salmonella spp. and hepatitis A/B/C viruses.
• Access to the lab is restricted, and procedures that create aerosols are conducted in biosafety cabinets (BSCs).
• Required safety equipment includes PPE (lab coats, gloves, and face shields) and Class I or II biosafety cabinets.
• An autoclave for waste decontamination and self-closing doors are required.
• Typically represented in clinical laboratories.

Biosafety Level 3 (BSL-3)

• Involves high-risk agents that can cause serious or potentially lethal disease through inhalation.
• Examples include pathogens that cause severe disease, such as Mycobacterium tuberculosis and SARS-CoV-1.
• Strict access control and medical surveillance for personnel are mandatory.
• All work is conducted in biosafety cabinets or other physical containment devices.
• Required safety equipment includes enhanced PPE, such as respirators and full-body suits, as well as Class II or III biosafety cabinets.
• Requirements include double-door entry (airlock) and HEPA filtration for exhaust air.
• Commonly found in research laboratories studying tuberculosis or SARS-CoV-1.

Biosafety Level 4 (BSL-4)

• Involves maximum-risk agents that are life-threatening, with no available vaccines or treatments.
• Examples include exotic pathogens that cause severe or fatal disease, such as the Ebola virus and Marburg virus.
• Strict access control and extensive training for personnel are required.
• Full-body, air-supplied positive pressure suits are worn, and all work is conducted in Class III biosafety cabinets or isolated rooms.
• Full-body, air-supplied suits with independent breathing apparatus, Class III biosafety cabinets, and an isolated building or zone within a building are required.
• Double-HEPA filtration for exhaust air is also required.
• Represented in laboratories studying Ebola virus or other hemorrhagic fever viruses.