RE-ECHO: BIOSAFETY IN THE LABORATORY

By: Ms. Lalaine Ilagan,RMT Ms. M.C. Plecis, RMT

Definition of Biosafety
Safety from exposure to Infectious Agents Principles and practices employed to protect laboratory personnel and the environment from exposure or infection while working with living organisms, biological materials, or agents. Promoting safe laboratory practices, and procedures; proper use of containment equipment and facilities; provides advice on laboratory design and risk assessment of experiments involving infectious agents and others sources
Bottom Line: Risk & Containment

Principles of Biosafety
Containment The purpose of containment is to reduce or eliminate exposure of:
laboratory workers, other persons the environment

Safe methods for managing infectious agents in the laboratory Environment

The location where the agent being handled, maintained or stored.

Evolution of Biosafety Programs

Biosafety program were established: To promote the use of safe microbiological practices, safe equipment and facility safeguards To reduce Laboratory-Acquired Infections (LAIS) To protect the public health and environment

Laboratory Associated Infections (LIAs)

Either symptomatic or asymptomatic, Acquired through laboratory or laboratoryrelated activities A result of working with infectious agents

What causes LAIs?

Improper Facility Design 1% 9% 90%

Containment Equipment Failure Improper Laboratory Practices -

Common problems
Bad practice in laboratory management Poor supervision of less experienced professionals Lack of accountability for occupational health and safety Lack of biosafety policy Lack of biosafety procedures and staff training in biosafety practices Lack of internal and external quality assurance

Agents and Risks

The agent is the what creates risk Risks to the worker or environment are often unknown

Assessing Risk

There is always risk!

The risk must be identified The risk is evaluated The risk must be measured Plan to minimize the risk

Risk assessment
Serves as a guide for selecting appropriate biosafety levels and microbiological practices, safety equipment, and facility safeguards that can prevent LAIs. Allows prioritization of engineering controls Use the hazard rating to justify resource allocation

Steps in Risk Assessment Process

Identify agent of potential hazard Identify hazards in laboratory procedure Determine the appropriate biosafety level and select additional precautions indicated by the risk assessment Evaluate the proficiencies of staff regarding safe practices and the integrity of safe equipment Review the risk assessment and the biosafety professional, safety matter expert and IBC (International Biosafety Committee)

Who Determines Acceptable Risk?

Assessment is conducted by a Biosafety Professional in partnership with and based on information provided by the Principal Investigator The assessment is presented to the Institutional Biosafety Committee (IBC) for approval

Identifying Risk
Understand the biology of the agent Susceptibility and transmission within the host Hazards associated with equipment and procedures Goal:
Provide the highest practical protection and the lowest practical exposure

Evaluating Risk Acceptability

Worst case scenario -What might happen? Likelihood of an event Seriousness of the incident

Actions needed to resolve the problems

What is Acceptable Risk?

Since there is no such thing as no risk Safe means risk has been judged acceptable Judging risk is a subjective- humans make decisions Measuring risk is objective- use available guidelines, data, and documentation Keep records of how determinations were made due to subjective nature of the process

Hazard, Threat, Risk

Hazard is a source of a potential for causing ; Hazard is not a risk without a specific environment A threat is a person who has an intent and or ability to cause harm Risk is the likelihood of an undesirable event, involving a specific hazard, that has consequences; risk can be based on either a hazard and or threat

Hazard Recognition
Knowledge of agent characteristics Risk assessment Sources of assistance

Understanding How Laboratory Associated Infections (LAIs) Occur

The Chain of Infection
Pathogenic agent

Susceptible Host

Reservoir

Portal Entry

Means of Escape

Route of Transmissio n

Risk Groups( RGs) Classification of pathogens

Reference: WHO-World Health Organization

RG-1
low individual or community risk Unlikely to cause disease in humans or animals

RG-2
Moderate individual risk, low community risk, May cause disease but typically not serious (Lab exposures can cause disease but unlikely to be a serious hazard); treatable

RG-3
High individual risk, low community risk ( serious respiratory agents )
May cause serious disease, usually treatable

RG-4
- High individual and community risk (Easy transmission)
- Serious or fatal; can be easily transmitted ; often not treatable

Pathogenic agent

Activity:
Instruction: Identify Risk Group Classification of various pathogens
Pathogens: RG -1: No or low individual and Community risk RG-2: Moderate individual risk, low community risk HIV E. Coli Answers: E. coli

HIV
Rabies, MTB Ebola

RG- 3: High individual risk, low community risk

RG-4: High individual and community risk

Ebola
Rabies MTB

Laboratory Reservoir
Blood units Blood Samples Laboratory workers Equipment

Reservoir

Means of Escape
Accidental releases and spills Incidental releases - manipulation of samples - aerosolization during sonication or centrifugation - Excretion of agents by animals
Means of Escape

Incident 1 A definite and separate occurrence; an event. See Synonyms at occurrence. 2 A usually minor event or condition that is subordinate to another. 3 Something contingent on or related to something else. 4 An occurrence or event that interrupts normal procedure or precipitates a crisis: an international incident. Accident 1 An unexpected and undesirable event, especially one resulting in damage or harm: car accidents on icy roads. 2 An unforeseen incident: A series of happy accidents led to his promotion. 3 An instance of involuntary urination or defecation in one's clothing. 4 Lack of intention; chance: ran into an old friend by acciden

Mode of Transmission
Aerosol Injection Ingestion Indirect contact

Route of Transmissio n

Element of Containment
Practice and Technique

Safety Equipment

Facility Design

Components of Laboratory Safety

Primary Containment Practices --technique --risk assessment Engineering Controls -- PPE -- Sinks -- Biosafety Cabinets -- Locks Secondary Containment -- Facility

The purposes of containment are to reduce or eliminate exposure of laboratory workers, other persons, the environment and; safe methods for managing infectious agents in the laboratory

 Primary containment
Protection from exposure to infectious agents It is provided by both good microbiological technique and the use of appropriate safety equipment.

Secondary containment
Protection of the environment external to the laboratory from exposure to infectious materials Facility design Operational practices

Three elements of containment include:

1. Laboratory practice and technique, 2. Safety equipment 3. Facility design

The risk assessment of the work to be done with a specific agent will determine the appropriate combination of these elements.

Laboratory Practices and Technique

Strict adherence to good microbiological practices and technique (Aseptic Technique) Sufficient training of all employees is provided by the supervisor Hazard awareness and standard operating procedures (SOP) established to manage them

Standard Laboratory Practices

Hygiene Rules

Eating, drinking, smoking, applying cosmetics or handling contact lenses not permitted in laboratory working areas Wash hands after working with agents and before leaving laboratory Do not recap needles

Steps in Proper Hand Washing

Remove jewelleries bare below elbows Wet hands and apply soap: 1. Rub palm to palm 2. Rub back of hands with palm of the other hand, fingers interlaced 3. Rub palm to palm with fingers interlaced 4. Rub back of fingers with hands clasped 5. Rub thumbs 6. Rub tips of fingers into palms 7. Rub wrist Wash soap away and dry thoroughly

Donning and Doffing Order of PPEs

Donning: 1. Gown 2. Mask 3. Goggles 4. gloves Doffing: 1. gloves 2. goggles 3. gowns 4. mask

Decontamination
Decontamination is a term used to describe a process or treatment that renders a medical device, instrument, or environmental surface safe to handle Sterilization is the use of a physical or chemical procedure to destroy all microbial life, including highly resistant bacterial endospores Disinfection eliminates virtually all pathogenic non-sporeforming microorganisms but not necessarily all microbial forms or inanimate objects Antisepsis is the application of a liquid antimicrobial chemical to skin or living tissue to inhibit or destroy microorganisms. It includes swabbing an injection site on a person or animal and hand washing with germicidal solutions

Categories of Decontamination
1. Heat Dry Wet Incineration 2. Liquid disinfection 3. Vapor and Gases 4. Radiation

Liquid disinfection
All purpose lab disinfectant Biohazard spillages Domestic bleach:
1:50= 1g/l== floors 1: 10= 5g/l ==blood spills Contact time: 10 minutes

= 1g/ L = 5 g/L =50 g/L

http://www.cdc.gov/od/ohs/pdffiles/ Module%202%20-%20Biosafety.pdf

http://www.cdc.gov/od/ohs/pdffiles/ Module%202%20-%20Biosafety.pdf

http://www.cdc.gov/od/ohs/pdffiles/ Module%202%20-%20Biosafety.pdf

http://www.cdc.gov/od/ohs/pdffiles/ Module%202%20-%20Biosafety.pdf

http://www.cdc.gov/od/ohs/pdffiles/ Module%202%20-%20Biosafety.pdf

http://www.cdc.gov/od/ohs/pdffiles/ Module%202%20-%20Biosafety.pdf

http://www.cdc.gov/od/ohs/pdffiles/ Module%202%20-%20Biosafety.pdf

http://www.cdc.gov/od/ohs/pdffiles/ Module%202%20-%20Biosafety.pdf

http://www.cdc.gov/od/ohs/pdffiles/ Module%202%20-%20Biosafety.pdf

http://www.cdc.gov/od/ohs/pdffiles/ Module%202%20-%20Biosafety.pdf

http://www.cdc.gov/od/ohs/pdffiles/ Module%202%20-%20Biosafety.pdf

http://www.cdc.gov/od/ohs/pdffiles/ Module%202%20-%20Biosafety.pdf

http://www.cdc.gov/od/ohs/pdffiles/ Module%202%20-%20Biosafety.pdf

http://www.cdc.gov/od/ohs/pdffiles/ Module%202%20-%20Biosafety.pdf

http://www.cdc.gov/od/ohs/pdffiles/ Module%202%20-%20Biosafety.pdf

http://www.cdc.gov/od/ohs/pdffiles/ Module%202%20-%20Biosafety.pdf

Biosafety Levels
Biosafety levels (BSL) BSL1 BSL2 (Some agents and materials requireenhanced precautions*) BSL3 References

A. Hazard levels
1. Degree of hazard Low risk: Well characterized agents not known to cause disease in healthy adult humans Moderate: Agents that cause human disease of moderate hazard High: Agents that cause disease of moderate to high hazard that have serious or potentially lethal consequences Biosafety in Microbiological and Biomedical Laboratories, Centers for Disease Control and Prevention (CDC)

2. Examples

Escherichia coli (laboratory strain)

Listeria monocytogenes

Mycobacterium tuberculosis

B. Standard microbiological practices Biosafety levels (BSL) BSL1 BSL2 (Some agents and materials requireenhanced precautions*) BSL3

1. Access to the laboratory

Access does not have to be restricted however, doors cannot be propped open (in violation of fire code).

Doors to the laboratory are closed when BSL-2 work is being conducted to prevent public access.

Doors to the laboratory are closed and locked to prevent untrained personnel access.

2. Biohazard signage 3. Biohazard solid waste decontamination

No biohazard sign is required. Biomedical waste vendor.

Biohazard sign must be posted. Biomedical waste vendor or steam sterilize with EH&S approval. 10% bleach/water made fresh daily with bleach having an EPA registration number (e.g., Chlorox) for 30 minutes or steam sterilize with EH&S approval. Permitted only in designated clean areas.

Biohazard sign must be posted. Steam sterilize in laboratory EH&S may grant exceptions in extenuating circumstances. Steam sterilize in laboratory EH&S may grant exceptions in extenuating circumstances.

4. Biohazardous liquid culture decontamination

10% bleach/water made fresh daily with bleach having an EPA registration number (e.g., Chlorox) for 30 minutes.

5. Eating, drinking, application of cosmetics or contact lenses 6. Contaminated sharpes (e.g., needles, blades, glass)

Permitted only in designated clean areas.

Not permitted at any time.

Safe handling practices must be developed and implemented. Substitute plasticware for glassware whenever possible.

7. Decontamination of work surfaces

8. Pipetting

Daily, after finishing work and following spills.

Mechanical device no mouth pipetting.

D. Laboratory facilities Biosafety levels (BSL) BSL1 BSL2 (Some agents and materials requireenhanced precautions*) BSL3

1. Ventilation 2. Handwashing facilities

Negative pressure. No recirculation or exhaust air to other areas of the building is permitted. Required. Required (foot/elbow/electronic operation) Required in laboratory. Required. Required in laboratory.

3. Autoclave 4. Eyewash station

Not required. Recommended. However, use of hazardous chemicals may change this to a requirement. Required.

5. Doors

6. Chairs 7. Cleaning and decontamination

Required. A series of 2 selfclosing doors is the basic requirement for entry. The space between the 2 doors is called the anteroom. Palm scanner are used to restrict access. Chairs used in laboratory work must be covered with a non-porous material that can be easily cleaned and decontaminated with appropriate disinfectant. Laboratory design should allow the facility to be easily cleaned and decontaminated. Carpets and rugs are not appropriate. Laboratory design should allow the facility to be easily cleaned and decontaminated. Carpets and rugs are not appropriate. Seams, floors, walls, and ceiling surfaces should be sealed. Spaces around doors and ventilation openings should be capable of being sealed to allow for space fumigation.

Required. Doors should be self-closing and have locks.

BLS-4:
Dangerous or exotic, high risk agents Life threatening disease, no vaccine or therapy Examples: Ebola. <arburg, Lassa, Machupo viruses

WASTE MANAGEMENT
The collection, transport, processing, recycling or disposal, and monitoring of waste It is generally undertaken to reduce their effect on health, the environment or aesthetics Carried out to recover resources Can involve solid , liquid, gaseous or radioactive substances with different methods and fields of expertise for each

Types of Wastes
1. 2. 3. 4. Non-infectious waste Infectious waste Highly infectious waste Sharp waste

Waste Handling
A system of marking, coding or identification should be understood by staff, especially for personnel engaged in the collection of waste for disposal off-site so that appropriate handling and disposal technique are employed Appropriate codes of practice must be enforced at both ends of the waste disposal system

Pre- treatment Requirements

Biological waste disinfection with the use of disinfectants and autoclaving Sharps- autoclaving or application of heat Chemical waste- storage compartment is necessary and the application of delay to decay process must be adopted

Collection and Transportation System

Disposal System