Basics of Biosafety

Working Safely with Biological Materials

Principles and practices employed to protect laboratory personnel and the environment from exposure or infection while working with living organisms, biological materials, or agents.

Included are any materials that may be potentially infectious.

Includes recombinant DNA research

What is Biosafety?

Hierarchy of Controls

Administrative Control: Rotating workers more often to reduce exposure times

Engineering Control: Installing guards on machinery, ventilation, sound proofing

Safe Work Practices: policies and procedures increasing safety either by changing the actual way the work is done or by adding a tool to help

Personal Protective Clothing or Equipment: respirators, ear plugs

The Hierarchy of Controls is the preferred method of controlling preventing and controlling hazards. You must document that you went through this process.

permissible exposure limit

Administrative Controls Authorization/Approval Written biosafety procedures required for the

experimental procedures and equipment including inventory of biological agents or materials

Laboratory personnel biosafety training Medical Surveillance (BSL 2 and above)

Health historyMedical screening ImmunizationSerum storagePost-exposure prophylaxis

Engineering Controls

Biological safety cabinets, glove boxes Animal containment caging systems Safety equipment (filtered or sealed

equipment) Ventilation system Containment facilities

Personal Protective Clothing and Equipment

Provides barrier against skin, mucous membrane or respiratory exposure to infectious agents during procedures

Prevent spread of contamination Integrity wanes with use (i.e., change

gloves frequently)

BSL are used in risk management

BSL are ways to control the agent facilities, safety equipment, practices, PPE, etc.

Once risk is assessed then the appropriate BSL is determined

Biosafety Levels (BSL)

Well characterized, non-pathogenic organisms or agents

Open bench- no containment

Use good laboratory practices, waste disposal, and aseptic techniques (i.e. performed under sterile conditions)

Example: E. coli K-12 strains

BioSafety Level 1

BSL 1: Work Practices and Procedures

ApplicationsNon-infectious agent and tissue culture,

media preparation Prevent Cross Contamination

Keep cultures coveredFlame instruments and containersUse sterile media and equipmentKeep hands or face away from cultures

Biosafety ProceduresWork with agents may be conducted on

open benchWash hands oftenNo mouth pipettingNo eating or drinking in labMinimize aerosol generationDecontaminate work surfacesWear applicable PPE

BSL 1: Work Practices and Procedures

Agents of moderate hazard to personnel or environment

Basic lab, but restricted access, containment during certain processes (i.e. aerosols, large volumes, etc.)

Autoclave and Biological Safety Cabinet desired

Use good laboratory practices, waste disposal, and aseptic techniques

Example: most non-respiratory, non lethal, agents

BioSafety Level 2

BSL 2: Work Practices and Procedures

Increasing emphasis on safety procedures and practices

Increasing need for staff training Increasing need for competent

supervision Biohazard sign posted at entry door Biohazard labels affixed on regulated

waste containers Use of personal protective equipment

as a barrier to exposure: lab coat, gloves, eye and face protection

Some work on open bench allowed

BSL 2: Work Practices and Procedures Aerosol generating procedures

performed in a biosafety cabinet: Homogenizing Vortexing Vigorous mixing Pipetting infectious liquids Sonication Pouring

If breach occurs: Evacuate lab, post spill sign With appropriate PPE and

disinfectant, decontaminate centrifuge, buckets, other items or areas

Agents of high hazard to personnel or environment

Respiratory exotic or indigenous agents which are easily transmissible causing serious or lethal disease

All work is contained, engineering controls and controlled environments

Example: Mycobacterium tuberculosis, SARS, etc.

BioSafety Level 3

FORGET ABOUT IT!!!

Hemorrhagic fever, deadly viruses, etc.

Total containment, airtight labs, “submarine” doors, air pumps, water treatment, etc.

Positive pressure “moonsuits”

BioSafety Level 4

Exposure Controls Careful Pipetting Techniques

Never blow out last drop in pipette Use pipetting aids with filters Never mix by suction and expulsion

(mix by sonication) Discharge liquid down side of

container, using tip-to-wall contact Deliver as close as possible to

contents Work over plastic-backed absorbent

matting (ensure it doesn’t slide forward or backward blocking air grill)

Exposure Controls Use Extreme Care with Sharps

Use sharps if only absolutely required as part of a process

Percutaneous exposure risk Employ safe work practices Utilize safe sharp devices

Aerosol exposure risk Use biosafety cabinet for

removal of air from needle Use mechanical methods for

needle removal Never bend, recap or manipulate

sharps by hand Keep hands away from needle

Exposure Controls

Safe Work Practices Wash hands after

each glove use and immediately or ASAP after exposure.

Remove PPE before leaving work area.

Flush body parts with water after contact with blood or OPIM

OPIM = Other Potentially Infectious Material

Exposure ControlsSafe Work Practices

Clean-up of spills and broken glassware/sharps contaminated with blood or OPIM

• Wear protective eyewear and mask if splashing is anticipated

• Remove glass and other sharps materials using a brush and dust pan, forceps, etc. Do not use your hands

• Properly discard all materials into a sharps or puncture-resistant biohazardous waste container

• Use paper/absorbent towels to soak up the spilled materials

Exposure ControlsPersonal Protective Equipment (PPE)

• Gloves

- Latex

- Nitrile

- Vinyl

- UtilityNitrile and vinyl gloves

Exposure ControlsPersonal Protective Equipment (PPE)

• Protective clothing

- Lab coat- Gown- Apron- Shoe cover or

boot

Exposure ControlsPersonal Protective Equipment (PPE)

• Eye-Face Protection and Masks

- Safety glasses with side shields

- Splash goggles

- Face shield- Mask

Biohazardous/Medical Waste Waste that is potentially infectious to humans, animals or plants

Biohazard Labels and Signs:

• Must have biohazard symbol• Labels attached securely to any

containers Biohazardous /Medical waste

Predominantly fluorescent orange

or orange/red background

Lettering and symbol in contrasting color

to background

Biohazardous Waste Categories

Cultures and stocks of infectious agents and associated biologicals

laboratory wastebiological production wastediscarded live and attenuated vaccinesculture dishes and related materialscontaminated PPE

Biohazardous Waste Categories

Liquid human and animal waste liquid or semi-liquid blood and blood

products and body fluidscontaminated items that would release

blood or items that are caked with blood or other potentially infectious materials; NOT including urine or materials stained with blood or body fluids

infectious animal waste (research)

Biohazardous Waste Categories

Pathological waste tissuesbody parts other than teethproducts of conception fluids removed by trauma or during

surgery or autopsy/necropsy or other medical procedure and not chemically fixed.

…And More Biohazardous Waste Categories

Animal and plant pathogen waste Recombinant DNA waste Sharps

Managing Liquid Biohazardous Waste

Storage: Label and secure bulk

vessels if not disposed of immediately

Treatment: Chemical disinfection OR AutoclaveDisposal: THEN Flush to sewer Use proper PPE!

Managing Non-Sharp Biohazardous Waste

labeled container

lined with a biohazardous waste bag

equipped with a lid.

Managing Non-Sharp Biohazardous Waste

Securely tie bags for transport to treatment/collection site.

When moving wastes, use secondary containment; avoid using public halls and elevators.

“Breakable” Non-sharps Biowaste

Store in labeled containersthat are puncture-resistant, closable and will capture leakage, BUT….

…Do NOT use SHARPS containers!

#3Is it a sharps hazard?

Examples:– needles– syringes– Scalpels مشرط– all biologically contaminated objects that

can easily penetrate skin (Pasteur pipettes,razor blades, etc.)

Place sharps in approved sharps container for disposal!

…Syringes in research settings should be disposed of as a sharp to avoid public relations concerns!

Sharps Containers Containers must be leak-proof,

puncture-resistant, closable & labeled with the biohazard symbol.

Proper sharps containersmust be used forboth clinic andfield work.

Proper Use of Sharps Containers

Place tops on containers before use on lab bench

Don’t forget to date the container when first put into use

Remember: sharps containers are aone-way disposal system

Proper Use of Sharps Containers

Use sharps containers for sharps ONLY!• No solid biohazardous waste (i.e.

gauze, un-broken pipettes, gloves)

• No mercurythermometers

What’s wrong with this picture?

Sharps Container Disposal Containers must be permanently closed

and disposed:Within 90 days

of first useWhen ¾ full

Disposal methods:Landfill مكب Incineration الحرق We use waste hauler للنفايات متعهد

Safety Notes on Sharps Use Do not re-cap sharps Keep sharps container in

close proximity to point of use (i.e. limit handling) for easy disposal

Do not leave needles in pockets of coveralls or smocks

Managing All That Other Waste…

Do NOT discard medications in the trash.

Return to source for disposal or seek assistance.

Recordkeeping

Medical Records• For employees with

exposure• Confidential• Hepatitis B vaccination

status • Post-exposure evaluations• HCP’s written opinions• Information provided to HCP

as required

HCP = Health Care Property Investors

Recordkeeping

Training Records

• Dates• Content summary• Trainer name &

qualifications• Attendee’s names & job

titles