Transcript Refresher Biosafety PowerPoint

LABORATORY
SAFETY
Lisa Bognar Phillips
[email protected]
Risk Management & Safety
631-5037
http://www.nd.edu/~riskman
Including Emergency Response, Personal Protective Equipment and Hazardous
Waste Management Training
Chemical Hazards
There are two general hazard classes that
must be evaluated when assessing the
safety of a chemical:
 Physical Hazards
 Health Hazards
Chemical Hazards: Physical Hazards
 Compressed Gas - cylinders of compressed gas are
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under intense pressure.
Corrosivity – strong acids and bases can cause visible
destruction of metals at the site of contact.
Flammability - both solid and liquid chemicals can
serve as fuel sources to support a fire.
Reactivity - many substances will react violently if
allowed contact with air, water or other chemicals.
Friction, shock, light or heat can be enough to
trigger some reactions.
Oxidizability – compounds that release oxygen as
they decompose. React with flammables
Chemical Hazards: Health Hazards
 Irritants - cause reversible inflammation of skin, eyes or
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nasal passages.
Sensitizers - cause allergic reaction after repeated
exposures. Individual sensitivities vary.
Carcinogens - alter DNA or cause cells with altered DNA
to multiply.
Systemic Toxins - typically target a specific organ or
system (liver, kidney, blood, nervous system,
reproductive system, etc.).
Toxins – based on dose. Anything can be poisonous.
Corrosives – cause burns to the skin, eyes, mucous
membranes and respiratory tract. HF is not like typical
mineral acids. It is extremely toxic and penetrates to the
bone. Calcium gluconate is the only antidote that should
be used on an HF burn.
Routes of Entry
 Inhalation:
The
major route of entry
when working with
solvents. Great rapidity
of absorption
Routes of Entry
 Absorption:
May
produce systemic poisoning.
Condition of skin determines
rate of entry. Examples:
organic lead, solvents, (xylene,
methylene chloride) organo
phosphate pesticides,
cyanides
Routes of Entry
 Ingestion:
Usually
accidental or due to poor
hygiene practices
Chemical Right to Know
.
MSDS Includes the following:
Product Information
Fire and Explosion
Toxicology
Health Effects
PPE
Storage
Leaks and Spills
Waste Disposal
First Aid
)
Product Information
Finding an MSDS
 Chemical manufacturers and distributors
CANNOT refuse a request for an MSDS.
 There are excellent on-line MSDS databases:
http://www.hazard.com/msds/index.php
http://riskmanagement.nd.edu/MSDS
University of Notre Dame
Chemical Hygiene Plan (CHP)
 Contains information and procedures that
laboratory personnel can use to protect
themselves from the chemicals they work with
 First place to look for answers to questions you
might have regarding chemical or laboratory
issues
 Located on Risk Management and Safety
website
 http://riskmanagement.nd.edu
 Topics include:
- Emergency Response - Personal Protective Eq.
- Chemical compatibility -Waste Management
- Standard Operating Procedures (SOPs)
Personal Protective
Equipment
 Gloves are required anytime you are handling
chemicals, biologicals and radioactive materials
 Eye protection
 Open toed shoes do not protect your feet and ARE NOT
allowed in the laboratory. (NO FLIP FLOPS, SANDALS,
etc)
Wear Eye Protection When Using:
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Glassware under reduced pressure
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Cryogenic materials
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Glassware under elevated pressure
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Explosives
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Caustics, Irritants or Corrosives
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Biohazards
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Radioactive Materials
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UV Light
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Toxic Chemicals
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Carcinogens
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Flammable Materials
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Lasers
Gloves
 Be sure you
know which
type of glove is
appropriate for
the chemical
you are using
Example of wrong glove having
been used
Gloves
Remove soiled
gloves. If gloves
become soiled or
contaminated
during experiment,
stop, and remove
carefully.
Wash your hands and put on new pair of gloves.
If you double-glove, remove soiled glove, replace
with new one, and continue your work.
Safety Equipment:
Chemical Fume Hood
Airflow into the hood prevents
chemicals inside from migrating
out into your breathing zone.
If air velocity into the hood is
impeded or slowed, the hoods
ability to capture chemicals is
compromised.
Factors that affect airflow:
•Sash Height
•Drafts
•Bulky objects inside
Gas Cylinder Safety
 Storage and Handling
 Gas cylinders should not be stored in exits or egress routes or
blocking any safety equipment
 Gas cylinders (excluding lecture bottles) should be stored in an
upright position and with safety caps in place unless in use.
 Use only the appropriate regulator for the gas
 Gas cylinders must be secured with a chain or appropriate belt
above the midpoint but below the shoulder of the cylinder
Cylinder Safety
What’s Wrong in
this Picture?
Cylinder must be secured by
chain or other steadying
device
Electrical Safety
 Eliminate frayed or worn
wiring
 Never stretch wires
across floor or other
equipment.
 Staff should know
location of circuit
breakers
Electrical Safety
 Match size of extension cord
to appliance power cord to
prevent cord overheating.
 Extension cords are not
intended for "permanent"
installations -- appliances
shall be connected to
permanently wired
receptacles.
Examples of Poor Storage
•Never store a chemical
with a missing or obscured
label
)
Container Labels
All chemicals (including solutions
and chemicals transferred
from their original containers)
should be labeled with their
common names,
concentrations and hazards.
Additional information such
as: date received, date
opened, date of expiration
should also be recorded.
Label may also include: Proper handling,
Storage, Emergency response
Refrigerators and Freezers
 Ordinary household refrigerators and
freezers constitute a hazard when used
for storage of flammable or unstable
chemicals. These units produce
conditions that can lead to explosions.
 Domestic refrigerators should not be
used for flammable chemical storage.
 "Lab-safe" refrigerators and freezers
(designed for storage of flammable
liquids) must be used for flammable
chemicals.
Definition
Biosafety
The application of combinations
of
laboratory practice and
procedure,
laboratory facilities,
and safety
equipment when
working with
potentially
infectious
microorganisms.
We use these practices to protect the workers and
those around, the product and the environment.
Laboratory Requirements
 Knowledgeable supervisor
 Knowledgeable personnel
 Aware of potential hazards
 Proficient in lab practices & techniques
 Availability of a lab specific biosafety
manual
 Contact information
 Copies of IBC,IRB,RCC, and/or IACUC
protocols
 Standard Operating Procedures
 For unique procedures
 Laboratory inspection checklist
 Emergency response plan
 Special procedures where needed

Relevant reference materials
Classification of
Infectious Agents
 Found in the NIH Guidelines.
 Classified into risk groups on the
basis of risk to the individual and to
the community.
 Currently, 4 risk group levels have
been designated.
 the least risk (RG-1) to the most risk
(RG-4).
RG-1 Agents
Agents
 Not associated with disease in
healthy adult humans.
 Generally only require a
laboratory with minimal
containment
 Biosafety Level 1 laboratory
 “Declaration of Dangerous
Goods” is not required for
shipment.
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Bacillus subtilis, Saccharomyces cerevisiae,
Staphylococcus epidermidis, Escherichia coli
K-12
RG-2
 Associated with human diseases
which are rarely serious.
 Preventative and therapeutic
interventions are available.
 Generally require a laboratory
with moderate containment.
 Biosafety Level 2 laboratory
 “Declaration of Dangerous
Goods” required for shipment.
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Measles virus, Salmonella spp., Toxoplasma
spp., Hepatitis B virus, leshmainia,
plasmodium falciparum
Biosafety Levels of Containment
 Combination of laboratory practices,
safety equipment, and laboratory design
to achieve levels of physical
containment.
 Currently, 4 biosafety levels have been
designated.
 The least restrictive (BSL-1) to the most
restrictive (BSL-4).
Containment:
BSL-1
 RG-1
2 Agents
BSL RG-2 Agents
 Not known to cause disease
 Associated with mild to
in healthy adult humans
moderate disease in
humans
 Practices
 Standard microbiological
practices
 Safety equipment
 None required
 Facilities
 Open bench top
 Practices
 BSL-1 plus limited access.
 Safety equipment
 Biological Safety Cabinet
and personal protective
equipment as needed.
 Facilities
 BSL-1 plus the availability of
a mechanism for
decontamination.
BSL-1 Standard
Microbiological Practices
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Restrict or limit access when working
Prohibit eating, drinking and smoking
Minimize splashes and aerosols
Decontaminate work surfaces daily
Decontaminate infectious waste
Maintain an insect & rodent control program
Prohibit mouth pipetting/ use mechanical
pipetters
 Needles & sharps precautions
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Use sharps containers
DON’T break, bend, re-sheath or reuse syringes or
needles
Use alternatives to needles when available
DON’T place needles or sharps in regular trash
containers.
BSL-1 Standard
Microbiological Practices
Wash hands
BSL-1 Safety Equipment
(Primary and Secondary
Barriers)
 Personal protective
equipment
 Eye wear
 Gloves
 Lab coat
 Countertops and floors easily
cleaned and decontaminated
 Laboratory Requirements:
 Lab does not need to be separated
from other lab sections and can be
of normal construction.

No specific ventilation req.
Training Requirements
BSL-1
BSL-2
 Supervisor

Trained in
microbiological or
related science
practices.
 Lab personnel

Specific training in
lab procedures.
 Supervisor
 Is a competent scientist
 Restricts access
 Lab Personnel
 Aware of potential hazards
 Proficient in practices and
techniques
BSL-2 Standard
Microbiological Practices
 Follow all practices and personal
equipment used for BSL-1
containment.
BSL-2 Safety Equipment
(Primary and Secondary
Barriers)
 Class II Biological Safety  Requirements:
 Laboratories have lockable doors
Cabinet recommended
 to protect product,
personnel, and the
environment.
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and separated from public areas
Sink for hand washing
Work surfaces easily cleaned
Bench tops are impervious to
water
Sturdy furniture
Adequate illumination
Air flows into lab without recirculation to non-lab areas
Windows fitted with fly screens
Biological Safety Cabinets
Barrier- Overview
 Protection of
 Product
 Personnel
 Environment
as Primary
 Class II Biological Safety
Cabinet
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Equipment is laid out to not restrict
airflow in the cabinet
“High efficiency particulate air”
(HEPA) filter
Traps particulates 0.01 microns and
larger including bacteria and viruses
 chemicals, fumes, and vapors
pass through
Biological Safety Cabinet - Operation
1. Load with needed supplies
2. Turn on and allow to run for 10-15
minutes
3. Check inward airflow with a piece of
tissue
4. Enter straight into cabinet and perform
work in a slow, methodical manner
5. At end of work package or decontaminate
items to be taken out of cabinet
6. Decontaminate interior of cabinet
7. Allow cabinet to run for 10-15 minutes
8. Shut off
9. Annual certification is required.
What happens when an
open flame comes in
contact with ethanol in a
Biosafety Cabinet
BSL-2 Facility Design
(Secondary Barrier)
Signage on the door with
restricted access when work in
progress or agents are present.
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Autoclave
available
Eyewash station
available
Decontamination Definitions
 Sterilization
 physical or chemical procedure used to destroy all
microbial life, including large numbers of highly
resistant bacterial spores.
 Disinfection
 physical or chemical procedure used to eliminate all
recognized pathogenic microorganisms but not all
microbial forms (bacterial spores) on inanimate
objects.
 Antiseptic
 germicide that is used on skin or living tissue for the
purpose of inhibiting or destroying microorganisms.
Decontamination: Conclusion
 Disinfectants do not replace
adherence to good standard
microbiological practices and
proper hygiene!
Biohazardous Waste
 stock cultures
 materials containing or contaminated with blood
 blood and body fluidsSerum,
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Plasma
Other blood components
Semen
Vaginal secretions
Cerebrospinal fluid
Pleural fluid
Peritoneal fluid
Pericardial fluid
Amniotic fluid
Any other body fluid visibly contaminated with blood
 sharps
 Pipette/pipette tips, Hypodermic needles, Scalpels, Breakable containers,
Glass products (i.e., slides or cover skips)
 animal waste
 other contaminated lab material
Biohazardous Waste
Containers
Biohazardous waste
containers shall be
clearly marked with
the universal
biohazard symbol.
•Puncture-proof, leak-proof,
sealable receptacles
•avoid over-filling
Orange Biohazard Bags
Orange Biohazard Bags contain
previously contaminated waste that
has been decontaminated
(autoclaved) by lab personnel. May
be taken to the dumpster if brown
bagged
Emergency Response
 Fires/accidents
911
(From a cell phone 574-631-5555)
 Spills
631-5037
(Risk Management and Safety during working
hours)
 After Hours Spills 911
State the type of emergency and the exact
location of emergency.
If a chemical is involved spell the chemical
name to the dispatcher.
Fire Safety
•Alert persons in area of fire
•Close doors to confine fire
•Activate fire alarm
•Evacuate through nearest
exit
•Do not use elevators
•Move away from the
building.
How to Use a Fire Extinguisher
pull pin
aim at the base of the fire
squeeze handle
sweep from side to side
Fire Extinguisher Training
Learn how to operate a
portable fire extinguisher
by participating in fire
extinguisher training
program sponsored by
Notre Dame Fire
Department. Contact
them at 1-6200 if you
would like to take the
class.
US&A (v 2/07)
Biological/Chemical Exposures
Needle Sticks:
Ocular Exposures:
Dermal Exposures:
•Wash thoroughly
with soap and water
•Rinse at eyewash at
least 15 minutes
•Identify source
(consent?)
•Hold eye open to
ensure effective
wash
•Rinse under drench
shower at least 15
minutes
•Remove all
contaminated clothing
•Seek Medical Treatment
•Your supervisor must fill out an accident/exposure form
First Aid - Chemical Burns (Eyes)
 Forcibly open eyelids to ensure
effective washing behind eyelid
 Wash from nose out to ear to avoid
washing chemicals back into eye or
into an unaffected eye
 Flood eyes and eyelids with
water/eye solution for
minimum of 15 minutes
First Aid - Chemical Burns (Eyes)
Make sure you know the location of the
nearest eyewash station.
Eye Wash Maintenance
 Eyewashes are periodically
checked by Facility Services,
but are not flushed. Lab
personnel must test eyewashes
monthly.
 Designate someone to test eye
washes monthly. Let run for 1
minute
Spill Response
 Be prepared for
emergencies
Biological
Spills/Decontamination
• Cover spill with paper towels
• Soak towels with disinfectant
(10% Bleach is effective)
• Wait 10 minutes
• While wearing PPE,
clean up the spill using
paper towels
Notify Risk Management
1-5037 if you need assistance • Wipe spill area with
disinfectant
• Dispose of all clean-up
materials as biological
waste (red bag)
Chemical Spills
Identify the chemical
Assess your ability to safely
contain and clean up the spill:
(spill size, chemical identification,
I CAN safely PPE)
I CANNOT safely
clean it up
Notify coworkers and
secure the area
Use spill kit to contain and clean
up the spill (MSDS helps)
Call Risk
Management to
confirm clean up
clean it up
Notify coworkers and
vacate/secure the area
Call 631-5037
During working hours
After hours 911
On cell call
574-631-5555
Chemical Waste Disposal
Satellite Accumulation
Container
•Compatible and in good condition
•Has “tight-fitting” closure and
container is closed unless adding
waste.
Label (3 things):
•Must be labeled as soon as first
drop of waste is put into the
container.
•Words “waste or “hazardous
waste”
•Description or Name of
chemical(s)
Secondary Containment
•Must segregate incompatibles
•One container per waste stream
Hazardous Waste Disposal Rules
 Do not use chemical
symbols or
abbreviations when
labeling your
containers
US&A (v 2/07)
341
Chemistry
Acetone
20
Methanol
40
4 Liters
Generator Signs
Faculty name printed
RM&S use
ONLY
Abbreviations and
formulas NOT acceptable
Person who generates
waste signs form
All other waste…
Gloves, paper, etc:
disposed of in regular (clear) trash
bags and bags are to be removed by
custodians daily.
Broken Glass: if clean, should be
disposed of in a puncture proof box
marked “Broken Glass”.
Laboratory Security
There are extremists who vandalize workplaces where they
suspect animal research is being conducted and others who
identify sources for chemicals/ biologicals to be used in
terrorism or illegal drug production
Protect your lab and your research:
• NEVER prop open any card or key
access corridor doors.
•Don’t let people ‘piggy-back’ to gain
access to restricted areas.
• Keep your lab doors closed and
locked when unattended.
•Question strangers, and report
suspicious activity to campus security.
Emergency Phone Numbers
• Security
911 or 631- 5555 from cell
• Risk Management & Safety 1-5037
• ND Fire
1-6200