Biological Laboratory

Safety
 Biocontainment labs across the Asia-Pacific
region all too often fail to live up to the term.

 An inspection of dozens of labs has found that

nearly one-third of the biosafety hoods intended to
protect workers from deadly pathogens did not
work properly
 Types of Laboratory Hazards
 Electricity hazards

 Gas Hazards

 Chemical Hazards

 Radiation Hazards

 Biological Hazards
 Biological Hazards
 Animals

 Blood

 Microorganisms

 GMO

 Transgenic Plants
 Safety Equipment (Primary Barriers)
 PPE

 Facility Design (Secondary Barriers)

 Biosafty
 Biosafety: Prevention of large-scale loss of
biological integrity, focusing both on
ecology and human health.

 All measures that prevent accidentally exposure

at biological agents (micro-organisms and
toxins) and infections and their released in
environment

See WHO Biosafety manual

Biohazard
Potential source of harm caused by biological
agents or toxins

Biological agent
Any microorganism including those which have been
genetically modified, cell cultures and endoparasites, which may be
able to provoke any infection, allergy or toxicity in humans, animals
or plants
 Regulatory Agencies
 The safety standards and practices described
here have been developed and are regulated by
government agencies including
 CDC (Center for Disease Control)
 NIH (National Institutes of Health)

 European Union Directive

 WHO (World Health Organization)

 IFBA (International Federation of Biosafety

Associations)
 Agenda
 Lab Safety
 Biohazards
 Biosafety Levels
 Biosafety Cabinets
 Lab Safety
 Safe Laboratory Practices
 Lab coats
 Safety glasses
 Proper footwear
 Hair back
 No food or drink in the
laboratory
 Biohazard
 An agent of biological
origin that can cause
disease in humans
 Microorganism
(Mycoplasma,
Mycobecterium)
 Toxin (cholra toxin)
 Allergen (Fungal)
 Biosafety
The combined use of
 laboratory practices,
 laboratory facilities and
 safety equipment
to work with potentially infectious
microorganisms.
 Why use biosafety practices?
To protect:
 Workers/Students

 Products/Experimental results

 Environment/Laboratory classroom
 Biosafety Levels
 BL-1: agents are not known to cause disease
 BL-2: agents are associated with human disease
 BL-3: agents are associated with human disease
and are potentially transmitted as aerosols
 BL-4: agents of life threatening nature
 Biosafety Level 1 (BL-1)
Use BL-1 when working with:
 Well characterized agents
 Non-pathogenic agents
 Agents that are not known to cause disease in health humans
 Agents that are of minimal hazard to lab personnel and the
environment

Examples of BL-1 Agents:

 E. coli JM109, DH5a
 Saccharomyces cerevisiae
 Biosafety Level 1 (BL-1)
Standard Work Practices
 Use mechanical pipetting devices
 Wash hands frequently
 Minimize splashes and aerosols
 Decontaminate work surfaces daily
 Handle wastes properly
 Maintain insect and rodent control program
 Biosafety Level 1 (BL-1)
Personal Protective Equipment (PPE)

 Lab coat or apron

 Safety glasses or goggles
 Gloves as needed
 Biosafety Level 2 (BL-2)
Use BL-2 practices when working with:
 Agents of moderate potential hazard to personnel and the
environment
 Pathogenic for humans
 Unlikely a serious hazard
 Treatment and preventive measures available
 Limited risk of spread of infection
Examples of BL-2 agents:

Human blood or body fluids

E. coli 0157:H7
Clostridium botulinum
Retroviral vectors
Human cells in cell culture
 Biosafety Level 2 (BL-2)
 Adequate illumination
 Eyewash facility
 Negative air pressure
 Autoclave available
 Biological safety cabinet
 Lab must be separated from public areas
 Biosafety Level 2 (BL-2)
Special Practices
 Place used slides and coverslips in sharps containers,
never in any other receptacle.
 Sharps containers are:
 Red in color

 Marked with the biohazard symbol

 Puncture resistant

 Leak proof
 Biosafety Level 2 (BL-2)
Personal Protective Equipment (PPE)
 Lab coat or apron
 Safety glasses or goggles
 Gloves
 Biosafety cabinet
 Aerosols or splashes

 Large volumes

 High concentrations
 Biosafety Level 3 (BL-3)
Risk Group 3 (high individual risk, low community
risk)
A pathogen that usually causes serious human or animal
disease but does not ordinarily spread from one
infected individual to another.
Effective treatment and preventive measures are
available.
Mycobacterium tuberculosis
 Biosafety Level 4 (BL-4)
Risk Group 4 (high individual and community
risk)
A pathogen that usually causes serious human or
animal disease and that can be readily transmitted
from one individual to another, directly or indirectly.
Effective treatment and preventive measures are not
usually available.
All kind of vireses
Ebola virus
Relation of risk groups to biosafety levels,
practices and equipment
Risk Biosafety Laboratory Laboratory Safety
group level type practices equipment
1 Basic – Basic teaching, GMT None; open
Biosafety Level 1 research bench work
2 Basic – Primary health GMT plus protective Open bench plus
Biosafety Level 2 services;diagn. clothing, biohazard BSC for potential
research sign aerosols
3 Containment Special As Level 2 plus BSC and/or
Biosafety Level 3 diagnostic, special clothing, other primary
research controlled access, devices for
directional air flow all activities
4 Maximum Dangerous As level 3 plus Class III BSC
Biosafety Level 4 pathogens unit airlock entry, shower Positive presure
exit, special waste suit + BSC II
disposal Double door
autoclave
Filtered air
 Risk group classification

Risk Individual risk Community risk

Group
1 no, low no, low

2 moderate low

3 high low

4 high high
 Biosafety Cabinets (BSCs)
 Provide product, personal and environmental
protection.

 Various classes of BSCs are available

 Amount of air exhausted
 Amount of employee protection
Biosafety Cabinets (BSCs)
 Biosafety Cabinets (BSCs)
 Sterile air is exhausted from BSCs through a HEPA
filter.
 HEPA filter can trap particles to 0.3 microns
 Chemical vapors and fumes pass through HEPA filters
 Biosafety Cabinets (BSCs)
Helpful Hints
 Enter straight into the cabinet, avoid sweeping
motions
 Don’t place materials on the grill
 Keep discard pan or bag inside the cabinet
 Decon materials before removal from the
cabinet
 Decontamination
 Sterilization: destroy all microbial life, including
spores
 Disinfection: destroy a majority of microbial
life, but not necessarily spores
 Antiseptic: germistat used on skin to inhibit
growth of microorganisms
 Decontamination
Agent Selection
 Degree of killing required

 Nature of item to be treated

 Ease of use

 Safety

 Cost

 Type and amount of organism

 Decontamination
Methods
 Heat: steam heat, dry heat, incineration

 Chemical: bleach, ethanol, hydrogen peroxide,

ethylene oxide, paraformaldehyde
 Radiation
 Decontamination

Disinfectants do not replace standard microbiological

practices and good hygiene!
 Biological Waste
Types
 Cultures, stocks

 Sharps

 Pipettes, tips and weigh boats

 Other materials
 Biological Waste
BL-1 Waste
 Solids
 Collect and package in clear autoclave bags
 Autoclave to sterilize
 Dispose of in regular trash dumpster (as long as no chemical
or radioactive contaminants are present)
 Liquids
 Collect in containers with lids
 Autoclave or treat with 10% bleach to sterilize
 Dispose of down the drain (as long as no chemical or
radioactive contaminants are present)
 Biological Waste
Sharps (needles, syringes, scalpel blades, slides,
blood vials, pasteur pipettes)
 Collect in approved sharps container

 Autoclave to sterilize

 Dispose of in medical waste boxes

 Risk Communication
 Your right to know

 What hazardous material you are working with

 Their hazards and risks

 How to protect from them