ACADs (08-006) Covered Keywords Laboratory safety, eye protection, protective clothing, storing chemicals, labeling chemicals, eyewash, showers. Description Supporting Material

An overview of Laboratory Safety Biosafety Dr. John Ridderhof Chief, Office of Laboratory Systems Development Division of Laboratory Systems/PHPPO

Presentation Outline Definitions General Safety Practices Laboratory Hazards – Biosafety Levels Decontamination Waste Disposal Safety Documentation Action Plan for Safety

Introduction Why Biosafety Practices? Protection: workers “products” co-workers lab support personnel environment 2.1

Safety practices should be applied throughout the testing process: Pre- analytical – Specimen collection – Specimen preparation – Specimen transport Analytical – Testing Post-analytical – Disposal

Examples of Laboratory Hazards  Growth of microorganisms Water baths  Aerosols, splashing, tube breakage Centrifuge  Accidental inoculation, aerosol, spillage Needles HazardOperation / Equipment

Safety Resources

Introduction Chain of Infection Reservoir of pathogen Portal of escape Transmission Route of entry/infectious dose Route of entry/infectious dose Susceptible host Susceptible host Incubation period Incubation period Risk Assessment PPE Immunization Surveillance Practices/ Equipment 2.1

Principles Biosafety Levels BSL1 - agents not known to cause disease. BSL2 - agents associated with human disease. BSL3 - indigenous/exotic agents with potential for aerosol transmission; disease may have serious or lethal consequences. BSL4 - dangerous/exotic agents which pose high risk of life- threatening disease. 2.1

Designing for Safety General requirements – Facility design – Water supply/sinks for hand washing – Ventilation – Standard lab practices Safety equipment – Personal protective equipment (PPE) – Biosafety cabinets

Biological Safety Cabinets 2.7 HAZARD ?

Biosafety Level 1 Standard Microbiological Practices Restrict or limit access when working Prohibit eating, drinking and smoking in the laboratory Pipetting by mouth strictly forbidden 2.3

Biosafety Level 1 Standard Microbiological Practices 2.3

Standard practices also include: Keep work areas uncluttered and clean No food in lab refrigerator Minimize splashes and aerosols Decontaminate work surfaces daily Maintain insect & rodent control program

Biosafety Level 2 Facility Design (Secondary Barriers) Requirements:  Laboratories have lockable doors  Sink for hand washing  Work surfaces easily cleaned  Bench tops are impervious to water  Sturdy furniture 2.4

Biosafety Level 2 Laboratory Facilities (Secondary Barriers) BSL-1 Facilities PLUS: – Autoclave available – Eyewash station available 2.4

Biosafety Level 2 Facility Construction (Secondary Barrier) Requirements: – Location - separated from public areas – Structure - normal construction – Ventilation - directional 2.4

Biosafety Level 2 Safety Equipment (Primary Barriers) In addition to BSL-1: Use biosafety cabinets (class II) for work with infectious agents involving: – Aerosols and splashes – Large volumes – High concentrations 2.4

Biosafety Level 2 Special Practices Needles & Sharps Precautions – Use sharps containers – DON’T break, bend, re- sheath or reuse syringes or needles 2.4

Biosafety Level 2 Special Practices Needles & Sharps Precautions (cont.) DON’T place needles or sharps in office waste containers 2.4

Biosafety Level 2 Special Practices Needles and Sharps Precautions (cont.) – DON’T touch broken glass with hands 2.4

Biosafety Level 2 Special Practices Identify “clean” and “contaminated” areas – Use appropriate warning signs Decontaminate work surfaces Report spills and accidents Remove gloves, lab coats before leaving work area No animals in laboratories 2.4

Biosafety Level 3 Laboratory Facilities (Secondary Barriers) BSL-1 and 2 Facilities PLUS: – Separate building or isolated zone – Double door entry – Directional inward airflow – Single-pass air; air changes/hour 2.5

Biosafety Level 3 Laboratory Facilities (Secondary Barriers) BSL-1 and 2 Facilities PLUS (cont.): – Enclosures for aerosol generating equipment – Room penetrations sealed – Walls, floors and ceilings are water resistant for easy cleaning 2.5

Biosafety Level 3 Safety Equipment (Primary Barriers) BSL-1 and 2 Safety Equipment PLUS: – BSC class II or III to manipulate infectious material 2.5

Biosafety Level 3 Safety Equipment (Primary Barriers) BSL-1 and 2 Safety Equipment PLUS: – Respiratory protection may be indicated 2.5

Biosafety Level 3 Special Practices BSL-2 Special Practices PLUS: – Work in certified BSC – Use bioaerosol- containing equipment – Decontaminate spills promptly 2.5

Biosafety Level 4 – Maximum Containment BSL -3 practices plus: – Clothing change before entering laboratory – Shower on exit – All materials decontaminated on exit from facility Safety Equipment: – Class III Biosafety cabinet – Class I or II biosafety cabinet – WITH full-body, air supplied, positive personnel suit

Biological Waste Types – cultures, stocks, isolates – materials containing or contaminated with blood – sharps – pipettes, wrappers, tips – All materials used in the lab

Specimen Disposal

Decontamination Sterilization Disinfection

Decontamination Definition Sterilization The use of a physical or chemical procedure to destroy all microbial life, including large numbers of highly resistant bacterial spores.

Decontamination Definition Disinfection The use of a physical or chemical procedure to virtually eliminate all recognized pathogenic microorganisms but not all microbial forms (bacterial endospores) on inanimate objects.

Decontamination Methods Heat Chemical Radiation

Decontamination Heat Types – Moist – steam – Dry – Incineration *The most effective method of sterilization

Decontamination Chemical Types – Liquids, i.e. chlorox, hydrogen peroxide – Gases, i.e. ethylene oxide

Decontamination Chemical General Lab Use - Hypochlorite Solutions – Large Spills/Large Organic Load undiluted from bottle – Small Spills/Virus Inactivation 10% - 1:9 – General Surface Disinfection 1% - 1:99

In case of a spill Wear disposable gloves Cover large blood spill with paper towels and soak with 1% (10000 ppm) of household bleach and allow to stand for at least 5 minutes Small spill - wipe with paper towel soaked in 1% bleach Discard contaminated towels in infective waste containers Wipe down the area with clean towels soaked in a same dilution of household bleach

Safety Documentation & Records Laboratory Safety Manual - Policies and Procedures Sample Contents: – Housekeeping – Personal protection – Safe decontamination of equipment – Decontamination & Waste Disposal – Emergency procedures  In-lab first aid  Accidental injury  Post exposure prophylaxis  Contacts – Personnel responsibilities Hygiene

Action Plan for Implementing Safety Practices Identify hazards Assess level of risks – Prioritize risk Establish and implement safety polices and procedures Conduct safety specific training – Must be a priority – Communication is key Perform regular audits and assessments

In Case of Exposure Be ready for the emergency before hand - Familiar with exposure specific policies - Conduct drills - Keep post exposure medicines available - Check periodically for stock and expiry of medicines Report immediately Go to the nearest, first available doctor

Post Exposure Write a report and reasons for accident Actions taken to avoid future accidents Training

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