ccm --bio,chem and animal - CCM Safety Training

Transcription

CCM
SAFETY MANUAL
--BIO, CHEM AND ANIMAL--
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BIO STANDARD OPERATING PROCEDURES
1. BIO-SOP#1 – PROPER USE OF A BIOLOGICAL SAFETY CABINET (BSC)
2. BIO-SOP#2 – POWER LOSS IN A BSC
3. BIO-SOP#3 – LABORATORY ENTRANCE/EXIT PROTOCOL (RM 2043 AND 1052)
4. BIO-SOP#4 – GUIDELINES FOR A BIOLOGICAL SPILL CLEAN-UP (SN#127)
5. BIO-SOP#5 – SAFE HANDLING OF LABORATORY COMPUTERS
6. BIO-SOP#6 – AEROSOL PRODUCTION AND EXPOSURE CONTROL
7. BIO-SOP#7 – PROPER USE OF A VACUUM IN THE LABORATORY
Center for Comparative Medicine
CCM Safety Manual (BIO-SOP #1)
Proper Use of Biological Safety Cabinet (BSC)
This SOP contains general information regarding proper BSC
use. Please refer to the manufacturer’s manual for additional
information regarding a specific BSC. Manuals for shared use
BSCs are located in 2002 CCM in the safety coordinator’s file
cabinet.
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General BSC information
1. The Biological Safety Cabinet (BSC), class II, is designed with
inward air flow at a velocity to protect personnel, highefficiency particulate air (HEPA)-filtered downward vertical
laminar airflow for product protection and HEPA-filtered
exhaust air for environmental protection.
2. Biological Safety Cabinets are among the most effective and
the most commonly used primary containment devices in
laboratories working with infectious agents.
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Specific Procedures
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1. Turn off the ultraviolet (germicidal) light and, if desired, turn on the fluorescent light. Never leave an
ultraviolet light on while there is anyone in the room. Turn off the ultraviolet (UV) light of all the BSCs in
the room where you work.
2. Open the glass front sash until the audible alarm no longer sounds.
3. Turn on the blower. Verify that you have cabinet airflow; either by
“ P” value
listening for blower sound or place a Kimwipe in the front opening (should
blow inward). The value on the magnehelic gauge (round with numbers
and a needle) or the air pressure monitor (vertical with lighted bars)
should match the “ P” value on the TSS certificate (square silver label).
Do not use the BSC if the reading deviates significantly in either direction
- report any significant deviations to the CCM Safety Coordinator.There
are also three 100% Exhaust BSCs (Class II B2) in CCM; only the one in
the shared BSL2 lab (2043A) is equipped with the gray “Fume Hood
Monitors” (see attachment #1). See the “BSL3 Practice and Facilities”
Power Point Presentation for information regarding the two 6 foot BSCs
are located in the BSL3 labs, 2042A&B. The “Fume Hood Monitor” must show a green light for ‘Normal
Standard Operation’. Do not use the BSC if the red light ‘Caution Flow Alarm’ is on (the audible alarm
may also be on or it may be muted).
4. Wipe down the interior area of the cabinet with a surface disinfectant. If the disinfectant is corrosive (i.e.
bleach), then clean the surfaces afterwards with a detergent and/or rinse with water. 70% ethanol is not
corrosive and may be used without rinsing with water.
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Special Notes:
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5. Place all materials to be used inside the cabinet. Disinfect the exterior of these materials. Implements
should be arranged in the cabinet’s work area in logical order so that clean and dirty materials are
segregated, preferably on opposite sides of the work area.
6. Blocking the front and rear perforated grills must be
avoided. If wipes or absorbent towels are used on the work
surface, be sure to keep them away from the grills.
7. After the BSC has operated for at least 3 minutes with the
window in the proper position, you are ready to begin.
8. Perform all work on the depressed area of the solid work
surface. Work with a limited number of slow movements.
9. Opening and closing doors in the laboratory causes airFront Grill;
disturbance which might interfere with cabinet airflow, this
Dokind
not
of activity should be kept to a minimum while the cabinetblock
is in grills
use.
10. Place pipettes in the BSC and discard into a suitable container inside the BSC to reduce the temptation
to move in and out of the work area unnecessarily.
11. Use good aseptic techniques. Procedures done with good technique and proper BSC methods will not
require the use of a flame. Flames disturb the directional air stream and also contribute to the heat load.
Gas lines may no longer be attached to BSCs without the approval of the campus Biosafety Officer. If you
currently have a flame, place it at the rear of the work area where the air turbulence caused by the flame
will have the least possible effect. Tubing for a burner within the cabinet should be resistant to cracking
or puncture, as well as UV light if left in the BSC with the UV light on.
12. After work is completed, disinfect all equipment and supplies before removing from the BSC and
decontaminate the entire work area with 70% ethanol or appropriate disinfectant. The BSC should then
be allowed to run for at least 3 minutes with no activity so the airborne contaminants will be purged from
the work area.
13. Turn off the blower, close the sash until the audible alarm no longer sounds, then turn on the UV light, if
you are the last to leave the room. UV lights are no longer maintained in the shared BSCs. BSCs may
be left with the blower running if the noise is acceptable.
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1. Sash position for 100% exhaust BSCs: Do not close the sash
completely for the three 6 foot 100% exhaust BSCs in rooms 2042A&B
and 2043. Leave the sash as open as possible without the alarm
sounding to allow the room to exhaust – note the “Leave a Gap”
signage. Closing the sash completely will restrict the room’s exhaust
airflow and could sound the Fume Hood Monitor (gray box) ‘Caution
Flow Alarm’ (red light) in 2043A or “Supply and Exhaust Alarm” in
2042A/B.
2. Chemical Use in BSCs: The HEPA (High Efficiency Particulate Air)
filter is one of the essential components of a BSC. It is the shield
between the environment and the experimental agent and is very
delicate. It must be pointed out that HEPA (and ULPA – Ultra Low
Penetration Air) filters do not filter gasses or vapors.
a. Recirculating BSCs: Any chemical that must be used in a chemical fume hood may not
be used in a BSC, as the chemical fumes recirculate to the room. Because BSCs
recirculate air, there will also be gaseous buildup to the point of equilibrium within the
BSC. An explosion can be the result of a motor spark or a burner operating in the work
area if chemicals used in the BSC are explosive or flammable.
b. 100% Exhaust BSCs: Intake air enters these BSCs (SterilchemGard) from above and
through the front opening. The air is totally exhausted. Because this 100% exhaust
feature permits no recirculation, the cabinet may be useful for work that generates small
amounts of chemical vapors and gases if these chemicals are known not to attack filter
components (glass fibers with corrugated separators). These 100% exhaust BSCs are
completely dependent on the building exhaust system –problems with the building
exhaust system will affect these BSCs.
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3. Remember these Important Operating Requirements for BSCs:
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a. Store equipment and supplies outside of the BSC!
b. Always turn on the blower (it really does happen – and not just to new
users!)
c. Always keep the air intake grills clear and unobstructed.
d. If the unit presents a warning signal, don’t operate the unit until the
warning ceases. Notify your supervisor and safety coordinator.
e. If you have a question, ASK! You are probably not the only one.
f. Minimize in and out movements when working!
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Power Loss in Biological Safety Cabinet (BSC)
 A loss of power to the BSC requires immediate attention to prevent accidental
exposure to the user and loss of sample sterility.
 Under NO circumstances should work continue with infectious materials
without power. The protective air barrier is lost during a power outage.
 See also “Proper Use of Biological Safety Cabinet (BSC)”: Standard Operating
Procedure (Exposure Control Plan) #9.
Protocol:
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1. When the power is lost to the BSC, immediately cover and/or close all
open containers, especially your infectious samples, including your media
and all medical waste containers.
2. Close the sash, leave your material in the BSC.
3. Turn off the blower for the BSC or the HEPA filter could be damaged when power returns.
4. If the outage continues for more than a short period, then move your closed cultures to the
CO2 incubator. Opening the incubator will cause the loss of 5% CO2 that will not be
replaced during the outage. However, if your cultures are stored in a water jacketed
incubator, they will be better maintained at 37oC than at room temperature in the BSC.
5. Avoid opening refrigerators and freezers during the outage. Material that must be
refrigerated, may be placed on ice (wet ice or dry ice, depending on the material).
6. When power returns, open the sash, turn on the BSC blower and wait 3 minutes (normal
start up) for the contaminated air to circulate through the HEPA filter. Check the magnehelic
gauge to measure pressure in the blower plenum. Resume normal activity.
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 The 6 foot 100% exhaust BSCs in 2042A&B and 2043A are on emergency
power, but…
1. The exhaust for these BSCs is connected to the building exhaust.
2. The building exhaust fans are also on emergency power.
3. However, the building exhaust fans TURN OFF! Continue, as above, for a power
outage – cover containers, close sash (leave the normal gap) and turn off the blower.
4. After approximately 5-10 minutes, the building exhaust fans will restart and airflow to
these BSCs will return. If the gray fume hood monitor (on the side in 2043A) or the
silver “Supply or Exhaust Failure Alarm” (on the front in 2042A&B) no longer shows a
blinking red light (the audio alarm may or may not be muted; attachment #1 and
photo on page or BSL3 Practice and Facilities PPP) and the Air Flow Monitor (AFM)
is green , then you may turn on the blower.
5. Wait 3 minutes (normal start up) for the contaminated air to circulate through the HEPA filter.
Activity may resume, if necessary; safe lab practice is to close materials and put them away
until normal power returns and the airflow stabilizes. The building supply fans are not on
emergency power, so the exhaust fans will be pulling air from every crack and crevice. You
may feel like you are in a wind tunnel and doors could be difficult to open or to close. Be
careful not to allow a door slam to on you!
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Laboratory Entrance/Exit Protocol
Documentation of initial and annual training required to work in shared laboratory 2043A and 1052.
See BSL3 Manual for entrance/exit protocols for the BSL3 laboratory (2042/A/B)
Communication with users in a shared lab is as important as PPE – be respectful of the ongoing work and
communicate with the users to share the space safely. Avoid motions that could interfere with procedures involving
infectious agents – especially the movement of agents within the lab (i.e., BSC to centrifuge, cultures from incubator
to BSC). Slow down!
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Personal Protective Equipment (PPE) required:
1. Upon entrance into 2043A and 1052 to check supplies:
A. If personnel are currently working in the space:
See #2 below and, remember, communicate!
B. If NO personnel are currently working in the space:
2 pairs of gloves are required to handle drawer knobs, etc.:
1 pair of inner gloves, donned in the anteroom*
1 pair of outer gloves, donned in the anteroom
2. Upon entrance into 2043A and 1052 to work:
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Closed-toe/heel shoes (bootie covers optional, but not a substitute for closed toe/heel shoes)
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Long pants (provided by user) or Tyvek pants (provided by lab group)
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Don (put on) in the anteroom, in the following order, prior to entrance into lab*:
1. One pair of inner gloves (long cuff preferred),
2. DuPont ProVent (or similar that resists penetration from blood and water-based liquids) gown, rear
closure preferred , with long sleeves (cuffs preferred); disposable sleeve covers optional
3. One pair of outer gloves, donned before/after gown; at least one pair of gloves must cover the gown
sleeve (no skin visible between gown and glove(s))
4. For splash protection: Face shield OR safety glasses AND mask
5. If hair is longer than shoulder length: Hair bonnet or other means of restraint (i.e., secured in “pony tail”
and tucked under gown)
3. Upon exit from 2043A and 1052:
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Doff (de-gown) PPE in the following order:
6. Check for overt contamination on your clothing while in the lab; any obviously contaminated clothing is
discarded in the medical waste container in the lab (could include your contaminated clothes!). –not in
the anteroom. If no overtly contaminated clothing is found, then proceed with doffing.
7. Remove outer gloves and disposable sleeves, if used, and discard in lab.
8. Lab 2043A: Back-bump the panic bar to avoid touching the “clean” door. Do NOT exit 2043A from the
rear door into the hallway. This exit is ONLY for extreme emergencies (i.e., earthquake, fire, personal
physical threat) and is labeled “For Emergency Use Only”.
9. Wearing your inner gloves, remove your gown, turning it inside out to avoid contact with outer surfaces
and store.
10. Remove Face Shield OR safety glasses AND mask (splash protection) and store.
11. Remove hair bonnet or booties, if used, and discard.
12. Remove inner gloves and discard in medical waste.
13. Wash your hands: use the sink in 2043; use alcohol rub before exiting 1052 and wash with soap and
running water as soon as feasible.
Additional comments:
 No HEADPHONES are allowed
 Use plastic containers to move cultures within the lab
 Use closed containers to transport cultures outside the lab
PPE to be provided by each principal investigator for their laboratory personnel
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*1052: “anteroom” is the don/duff “area” as there is no anteroom; “lab” is the working section (BSC, incubators, centrifuges, etc.)
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CCM Safety Manual
BIO-SOP #4
SafetyNet #127 – Biological and
Biohazardous Spill Response
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This Safety Net outlines the steps to take after a spill of any infectious agent or recombinant DNA
material has occurred in your laboratory or in nearby areas such as in a corridor. Although any
laboratory that uses hazardous materials is required to have an appropriate spill clean-up kit
available and to provide spill clean-up training, responding effectively and safely to a spill requires
judgment and risk assessment. If you are not comfortable with the situation or are not confident of
your abilities (even if you are thoroughly trained), or if you think that clean-up might entail
unacceptably elevated risk, discuss the spill with the Biological Safety Office staff at EH&S before
going further. No matter what action you decide to take, moderate to high-hazard spills as noted
below must be reported to the Biological Safety Office before you attempt to clean them up, and
under NIH and UC Davis rules all spills of all biological materials including spills of Risk Group
(RG) 1, RG2, or RG3 agents or any recombinant DNA materials must be reported to the Biological
Safety Office (530 752 1493) within one business day. You can report the spill by telephone or use
the online system at http://safetyservices.ucdavis.edu/programs/biosafety/biohazard-incident-report.
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This SafetyNet constitutes the standard UC Davis biohazardous spill response training document,
and includes a risk-related spill response matrix and a spill response instruction summary page
intended for laboratory posting. Before posting the matrix and instruction sheets please highlight
the matrix as appropriate to the types of biological agents handled in your laboratory.
Spill risk assessment: Evaluate the spill to determine the level of risk it represents, so that you can
decide whether you or anyone in your group has the training, knowledge, and equipment needed to
clean up the spill and to decontaminate all contaminated surfaces so that 100% of the spilled
material is removed or inactivated. Your risk assessment should also help you to determine whether
an immediate response with absorbent material is necessary to prevent the spill from seeping into
places that will be particularly difficult to clean. Consider:
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Biohazard potential of the spilled material (Risk Group (RG) classification, agent infectious
route, agent infectious dose)
Spill volume
Spill location
Extent of visible spatter (cryptic spatter is likely to be even more extensive)
Additional risks (e.g., does the spill include broken glass?)
Skill, experience, and health status of trained personnel
Availability of Personal Protective Equipment (PPE)
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Standard Operating Procedure #4 (Exposure Control Plan)
1. Moderate to high-hazard spills that must be reported to the Biological Safety Office before
clean-up but after necessary personal decontamination include:
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Any spill >500 ml
Any spill from a fermentor at Biological Safety Level 1--Large Scale (BSL1--LS) or above
Any spill in a Biological Safety Level (BSL) 3 laboratory
Any viable cultured RG2 agent of any volume outside a biological safety cabinet
Any viable cultured RG2 agent ≥10 ml inside a biological safety cabinet
Any spill of biological or biohazardous materials or agents in a publicly accessible area such
as a corridor
Spills of a RG2 or RG3 agent or rDNA construct inside a centrifuge that occurred during
operation, in an unsealed rotor or carrier
Spills of a RG2 or RG3 agent inside a refrigerator, especially spills discovered when the
door is opened
Any spill for which no person trained to clean up is currently available
Telephone number to call
Request assistance from:
530 752 1493
Biological Safety Office
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Spill time and location
Normal business hours
From the Davis or Sacramento
campuses
After hours and on weekends
From the Davis campus
From the Sacramento campus
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Biological Safety Office telephone contact information for immediate assistance:
911 dispatch
911 dispatch
EH&S 24/7 on call
EH&S 24/7 on call
Bleach or other approved disinfectant specific to your agents or materials
Spray bottle
Appropriate container to dilute disinfectant, if needed
Gloves (assorted sizes)
Eye protection/face shield and other appropriate PPE as noted below
Paper towels (at least one full package)
Long forceps or egg tongs (or both—egg tongs are better for picking up broken glass,
forceps may be better for pushing paper towels into tight corners, and for retrieving
disinfectant-soaked paper towels)
Red biohazard bags or clear autoclave bags, as appropriate for the spilled materials
Empty, appropriately marked sharps container for disposing broken glass (clear white
without biohazard label for RG1 materials, red with a biohazard label for medical waste
including human and non-human primate source materials and RG2 and RG3 infectious
agents)
A dust pan and brush for spills of dry RG1 material such as transgenic plants
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2. Spill kit: a biological or biohazardous spill kit should include the following items:
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Store these materials in a container of appropriate size (e.g. Nalgene tub, five-gallon paint bucket)
in an easily accessible location, and verify the integrity and completeness of the contents at least
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twice per year (ensure that the gloves are not degraded, that the disinfectant is not expired, that the
spray bottle, paper towels, sharps container, eye protection, and forceps have not been diverted to
other uses, etc). Be sure to label the container and the outside of the storage cabinet prominently.
To clean up a biological or biohazardous spill:
First Priority:
Assess yourself and other laboratory occupants for potential personal
contamination. If any personal contamination with a RG2 or RG3 agent or contaminated material is
found or believed to have occurred:
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a. Remove all contaminated clothing, quickly. Place contaminated clothing in a red
biohazard/autoclave bag to be autoclaved later. Do not contaminate public areas with
contaminated clothing. In anticipation of such emergencies, the PI should provide a
fire protection or other blanket that can be used to cover someone who must remove
biohazardous spill-contaminated clothing or who must use an emergency shower
following a chemical splash.
b. Flood the skin with flowing water for approximately 15 minutes and wash using soap
and water. Do not use hot water and do not scrub so vigorously that you abrade the skin.
c. If aerosol formation is believed to have been associated with the incident leave the
contaminated area immediately. Post the contaminated area to prevent entry until it is
safe.
d. Seek medical attention promptly: contact Occupational Health Services (530 752 6051)
and EH&S (530 752 1493). On weekends and after normal work hours call 911.
e. For eye splashes, hold the eyes open and irrigate with plenty of water at an eyewash
station for at least 15 minutes. Seek medical attention promptly: contact Occupational
Health Services (530 752 6051) and EH&S (530 752 1493). On weekends and after
normal work hours call 911.
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Second Priority: Clean up the spill:
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A. Wear appropriate PPE to clean spills (as detailed in the response matrix that
accompanies this SafetyNet).
B. If the spill involved broken glass, pick up the large pieces with the forceps or egg tongs
and dispose in a hard-walled sharps container. Handle broken glass with care!
C. Distribute paper towels around the periphery of the spill, then towards the center. Use
the forceps or egg tongs to push paper towels into recesses where spilled material may
have flowed.
D. Dilute your disinfectant to the appropriate concentration in a spray bottle (if available).
E. When the spill is fully covered with paper towels, spray or very carefully pour 10%
bleach or other approved disinfectant on the paper towels. Avoid generating further
aerosols or flooding the spill so much that untreated material may flow
F. Allow at least 30 minutes contact time.
G. Pick up the paper towels with large forceps or egg tongs and put them in the appropriate
waste bag. Change gloves and put used gloves in bag as well. Avoid direct contact with
the contaminated paper towels, even with gloved hands
H. Spray or carefully pour 10% bleach or other approved disinfectant on the surface
residue. Wipe up the residue with paper towels and place in appropriate bag. Small bits
and pieces of broken glass should be entrained in the wet paper towels and discarded
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I.
J.
K.
L.
M.
into the waste bag. Pieces too large or heavy to entrain must be discarded in a sharps
container.
Repeat step “H” at least once.
Seal and transport the waste collection bag to the appropriate autoclave or medical waste
accumulation site.
If broken glass was disposed in a sharps container, seal the container permanently,
decontaminate the exterior with the sprayed liquid disinfectant, and transport the sealed
container to a medical waste accumulation site or request a sharps pickup on the Safety
Services website (Davis campus)
Clean and disinfect the forceps or egg tongs and any other non disposable items before
returning them to the spill kit. If possible, autoclave the forceps or egg tongs before
returning them to the kit.
Report the spill to your supervisor and to the Biological Safety Office if you have not
already done so.
Guidelines and rules to help prevent spills:
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Practice manipulations involving biohazardous materials and agents by handling similar
volumes of non-hazardous materials with the same tools and containers in the same working
environment (e.g., biological safety cabinet) until you are adept and comfortable with the
entire procedure.
Always transport biohazardous materials outside of a biological safety cabinet in secure
secondary containment.
Always use sealed rotors or carriers to spin biohazardous materials in a centrifuge.
Always store biohazardous liquids in refrigerators in a manner that prevents spillage if the
container is tipped (secondary containment is important).
Always ensure that the bottom drain is closed before working at a biological safety cabinet.
Always transport biohazardous materials in publicly accessible areas in secondary leakproof
containment, with sufficient absorbent material to absorb the entire liquid contents of the
primary container. Label secondary containers with the universal biohazard symbol.
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Tips to help handle spills:
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Study the attached Spill Response Matrix in advance so that you know how to handle
location-specific spills.
Mark the dilution container in the spill kit in advance to show how much disinfectant to add
and how much diluent to add in addition, to avoid delays when the time comes to handle a
spill.
Keep a pair of shoes at the lab just for use in the lab. If you routinely change shoes when
you arrive at the lab and change back when you leave for the day you won’t track everyday
contaminants to your automobile or home, and if you need to remove your “lab” shoes
because of spill contamination you will still have shoes available to leave the lab.
Conduct periodic hands-on drills with volumes of spilled water similar to fluid volumes in
use in the laboratory to ensure that all laboratory staff members are well-experienced in the
location of the spill kit and in spill handling. Practice clean-up in typical and atypical spill
situations.
Created 06/08, IBC reviewed and updated 10/10 (nld, sjb) ; 4/16/13 update OccHealth#'s (kess)
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Biohazard Spill Response Matrix
University of California, Davis, Biological Safety Office, EH&S
530 752 1493
Highlight the rows that include the types of biohazardous materials your laboratory handles
Risk Group/Biological
Safety Level of
laboratory
Spilled Material
Spill Location,
Spill Volume
Where Applicable
RG1/BSL1
Microbial agents with no infectious or
pathogenic potential to humans or other
mammals; recombinant constructs, cloning
hosts, and non-infectious vectors, waste
materials such as spent culture media that have
been in contact with RG1 agents
All
Appropriate PPE
Lab coat, gloves, eye protection
Biological safety Lab coat or Tyvek gown, double gloves,
cabinet, <10ml
goggles or face shield
Conduct risk assessment
Deposit waste in clear autoclave bags or sharps
containers, autoclave the bags and dispose to
landfill, request a sharps pickup from Safety
Services, notify Biological Safety Office of incident
and clean-up results
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Conduct risk assessment1
Conduct risk assessment,1
notify Biological Safety Office
Evacuate the laboratory,
notify the Biological Safety
Office, conduct risk
1
assessment,
wait 30 minutes
Lab coat or Tyvek gown, double gloves,
before clean-up
goggles or face shield, and a surgical
mask or professionally fit-tested N95
respirator to entrain droplets
Discovered in
centrifuge or
refrigerator
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In public area
RG2/BSL2 aerosol
transmissible
pathogens
Enteric and viral RG2 agents designated in CalOSHA Standard 5199 App. D as potentially
aerosol transmissible pathogens;2 viral vector
preparations incorporating oncogene, toxin, or
virulence factor coding sequences; waste
materials that have been in contact with these
agents
Any
RG3/BSL3
All
Any
Deposit waste in red medical waste bags and
biohazard sharps containers, transport closed bags
to a medical waste accumulation site, permanently
close sharps containers and disinfect exterior
surfaces, request a sharps pickup from Safety
Services, notify Biological Safety Office of clean-up
results
Handle waste as described above for other RG2
agents, decontaminate the entire interior of the
unit, notify Biological Safety Office of clean-up
results, seek medical follow-up
Deposit waste in red medical waste bags and
Evacuate the area, divert foot biohazard sharps containers, transport closed bags
traffic, notify the Biological
to a medical waste accumulation site, permanently
Safety Office, conduct risk
close sharps containers and disinfect exterior
surfaces, request a sharps pickup from Safety
assessment,1 wait 30 minutes
Services, notify Biological Safety Office of clean-up
before clean-up
results
b
RG2/BSL2
disease in humans which is usually not serious
and for which treatments are often available,
viral vectors derived from agents capable of
infecting humans; plasmids that include coding
sequences for oncogenes, toxins, or virulence
factors, and other recombinant constructs
normally used at BSL2; waste materials such as
spent culture media that have been in contact
with RG2 agents or materials
Waste disposal and follow-up
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Biological safety
Human or non-human primate source materials cabinet, >10ml
such as established cell lines, primary cell
In laboratory,
cultures, tissues, blood, and body fluids,
outside of BSC
infectious or pathogenic agents that cause
Preliminary actions
Solid front lab coat or Tyvek gown,
double gloves, face shield or goggles,
and a professionally fit-tested
respirator (at least N95).
Evacuate the laboratory or
other area, divert foot traffic,
notify the Biological Safety
Office, conduct risk
assessment,1 wait 30 minutes
before clean-up
Handle waste as described above for other RG2
agents, notify Biological Safety Office of clean-up
results, seek medical follow-up
As determined and pre-approved by the Institutional Biosafety Committee
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Consider all risks that the uncontained agents or materials entail, determine whether your training is adequate to ensure complete clean-up of the spill and decontamination of all surfaces, determine whether an immediate response
such as immediate application of absorbent material is needed to prevent escalation of the spill hazard
2
e.g., Salmonella sp., Shigella sp., E. coli O157: H7, HIV in clinical samples, consult the Biological Safety website for the complete list
11/2/2010 sjb
Biohazardous Spill Clean-up
1. If this is a moderate to high hazard spill reportable to the Biological Safety Office
before clean-up (530 752 1493), have you reported it?
2. Have you confirmed that appropriate PPE is available?
3. Have you checked yourself and others nearby the spill for spatter or shoe
contamination?
4. Have you alerted the lab personnel and passersby (for spills in corridors) and
evacuated the lab if appropriate?
5. Have you located the spill kit and verified that you have everything you need?
6. For spills outside of the biological safety cabinet, have you allowed 30 minutes
settling time?
7. Are you trained in biohazardous spill clean-up?
15
If you answered “yes” to questions 1-7 and it is appropriate for you to clean up the
spill, you may proceed as outlined below:
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A. Wear appropriate PPE to clean spills.
B. If the spill involved broken glass, pick up the large pieces with the forceps or egg tongs
and dispose in a hard-walled sharps container. Handle with care!
C. Distribute paper towels around the periphery of the spill, then towards the center. Use the
forceps or egg tongs to push paper towels into recesses where spilled material may have
flowed.
D. Dilute your disinfectant to the appropriate concentration in a spray bottle (if available).
E. When the spill is fully covered with paper towels, spray or very carefully pour 10% bleach
or other approved disinfectant on the paper towels. Avoid generating further aerosols or
flooding the spill so much that untreated material may flow.
F. Allow at least 30 minutes contact time.
G. Pick up the paper towels with large forceps or egg tongs and put them in the appropriate
waste bag. Change gloves and put used gloves in bag as well. Avoid direct contact with
the contaminated paper towels, even with gloved hands.
H. Spray or carefully pour 10% bleach or other approved disinfectant on the surface residue.
Wipe up the residue with paper towels and place in appropriate bag. Small bits and
pieces of broken glass should be entrained in the wet paper towels and discarded into the
waste bag. Pieces too large or heavy to entrain must be discarded in a sharps container.
I. Repeat step “H” at least once.
J. Seal and transport the waste collection bag to the appropriate autoclave or medical waste
accumulation site.
K. If broken glass was disposed in a sharps container, seal the container permanently,
decontaminate the exterior with the sprayed liquid disinfectant, and transport the sealed
container to a medical waste accumulation site or request a sharps pickup on the Safety
Services website (Davis campus)
L. Clean and disinfect the forceps or egg tongs and any other non disposable items before
returning them to the spill kit. If possible, autoclave the forceps or egg tongs before
returning them to the kit.
M. Report the spill to your supervisor and to the Biological Safety Office
6
CCM Safety Manual (BIO-SOP#4a)
Guidelines for Biological Spill Clean-up
(based on the old version of SafetyNet #127)
This Safety Net outlines the steps to take after a biological spill has occurred. If you would like assistance or
recommendations for a spill clean-up you can call the EH&S front desk (752-1493), however you must call 911 (from a
campus phone) or 752-1234 (from a cell phone) if the spill is greater than 500 mls.
15
1. A Biological Spill Kit should include the following items:
• Bleach or other approved disinfectant
• Spray bottle or other appropriate container to dilute bleach
• Gloves (multiple sizes)
• Eye protection/face shield
• Paper towels
• Long forceps
• Red biohazard bags
• Broom and dustpan (to clean up broken glass)
20
2. The appropriate PPE should always be worn when cleaning a biological spill:
• Risk Group 1 agents – Lab coat or Tyvek gown and gloves
• Risk Group 2 – Lab coat or Tyvek gown, gloves, and at least a surgical mask
• Risk Group 2 agents that are aerosol transmitted (including plasmids with oncogenes) and all Risk
Group 3 agents – Lab coat or Tyvek gown, gloves, goggles, and a properly fit-tested respirator (at least
N95)
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3. To clean up a biological spill outside of a biosafety cabinet:
• Let the spill “settle” for at least 30 minutes (if it is a Risk group 2 or 3 agent), evacuate the laboratory,
and post signs on the doors to prevent re-entry before it is safe.
• Wear appropriate PPE mask to clean biohazardous spills outside of a biological safety cabinet.
• Distribute paper towels around the periphery of the spill, then towards the center.
• When the spill is fully contained, spray or pour 10% bleach (freshly diluted) or other approved
disinfectant on the paper towels, allow 30 minutes contact time.
• Pick up the paper towels with large forceps and put them in a red biohazard bag. Change gloves and
put used gloves in biohazard bag as well.
• Spray or pour 10% bleach or other approved disinfectant on the surface residue.
• Wipe up the residue with paper towels and place in red biohazard bag
• Repeat the last two steps at least once.
4. To clean up a biological spill inside of a biosafety cabinet:
• Always ensure that the bottom drain is closed before working at a biological safety cabinet.
• Use the same techniques described above regarding paper towel placement and disinfectant use, but a
30 minute wait for the initial spill to settle is usually not necessary unless some of the spill occurred
outside of the biological safety cabinet.
• If 10% bleach is used to decontaminate the spill on a stainless steel surface, follow with sprayed water
or 70% ethanol and wipe dry with paper towels.
For additional information on cleaning up a biological spill, contact EH&S at 530-752-1493, your EH&S Safety Advisor ,
or [email protected] . Report spills outside biosafety cabinet to your supervisor and the safety coordinator.
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CCM Safety Manual (BIO-SOP#5)
Safe Handling of Laboratory Computers
Staff personnel who service laboratory computers adjacent to research
materials or on research laboratory benches must be assured that the
equipment does not require special handling procedures.
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1. Computers must be free of chemicals, radioactivity and
biological reagents. If a computer is used with gloves as
standard lab practice, then the computer is considered
contaminated. The surface of the computer must be
decontaminated before any requested service is performed.
2. Computers on laboratory desks are considered
uncontaminated. Please notify staff personnel if this is not
the case.
3. Make every effort to locate computer equipment on
laboratory desks or tables that are not adjacent to research
activities.
4. If the equipment cannot be decontaminated, then the user
must directly supervise staff in safe handling procedures.
5. Keyboard covers are recommended for keyboards adjacent
to research materials to facilitate decontamination.
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Aerosol Production and Exposure Control
An analysis of laboratory-acquired infections (LAI) indicates that fewer than 20%
of known infections can be attributed to a documented accidental exposure. The
creation of aerosols is assumed to be primarily related to the remaining 80% of
these cases. Whenever work with infectious agents is performed, all appropriate
measures must be taken to protect workers and the environment.
15
Definition: Aerosols are liquid and solid particles suspended in the air. An aerosol with
a diameter of 5 microns or less can remain airborne for a long period of time; spread
wide distances, and is easily inhaled. Particles with a diameter larger than 5 microns
tend to settle rapidly and can contaminate skin, other surfaces and ventilation systems.
Examples of Aerosol-Producing Activities in the Lab:
20
Aerosol from blown
out pipette
Snap cap aerosol
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Blowing out pipettes
Cell sorters
Shaking tubes, vigorous stirring
Opening lyophilized cultures, opening snap top tubes, breakage of culture containers
Flaming loops or slides
Pulling needles out of septums, filling a syringe
Pouring liquids
Centrifugation steps such as filling centrifuge tubes, removing plugs or caps from
tubes after centrifugation, removing supernatant, resuspending sedimented pellets,
breakage of tubes during centrifugation and centrifugation itself
Sonicating, homogenizing, blending, grinding, cell disruption with French press
Intranasal inoculation of animals
Cage cleaning, changing animal bedding
Harvesting infected material from animals, eggs and other virology procedures
Necropsies of infected animals
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
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
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

Safe Work Practices to Minimize the Creation of and Exposure to Aerosols: Use a
combination of the appropriate safety equipment and safe procedures is the primary method to
minimize the creation of and exposure to aerosols.
Lab safety equipment to protect personnel from aerosols
 The certified biological safety cabinet (BSC) is the primary barrier to protect worker


from aerosols. Perform as many aerosol producing activities that involve infectious
agents as possible in a BSC. Other safety devices include safety centrifuges with
automatic locking mechanisms or solid lids, safety centrifuge cups, safety
blenders, safety sonicators.
If aerosol production cannot be prevented or contained, contact UCD
Environmental Health and Safety to determine if use of a respirator is appropriate.
Refer to your Biological Use Authorization for safety protocols specific to your
infectious agent.
Worker at Biological
Vacuum line trap and filter systems are used to protect the vacuum system from
Safety Cabinet
aerosols. See Exposure Control Plan SOP#13: Proper Use of Vacuum in the
Laboratory.
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Safe work practices for centrifugation of biohazards
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Centrifugation presents two serious hazards: mechanical failure
and dispersion of aerosols. A mechanical failure, such as a
disintegrated rotor, can produce not only aerosols but also
hazardous fragments moving at great velocity. These fragments,
if they escape the protective bowl of the centrifuge, could produce
traumatic injury to personnel and destruction of property. Although
accidents from improper use of centrifuges are far less frequent
than with pipettes or syringes and needles, when they do occur
and involve infectious agents, aerosols usually are created and the
Rotor Disintegration in
possibility of causing multiple exposures is considerably higher.
Ultracentrifuge

Routinely inspect centrifuge to
ensure that leakage is not occurring.
 Do not overfill centrifuge tubes or bottles. Wipe the outside of the tubes with
disinfectant after they are filled and sealed. Check your gloves for evidence of
leakage. DO NOT SPRAY YOUR GLOVES WITH ALCOHOL if they are
contaminated - replace your gloves - alcohol creates holes in latex disrupting the
barrier that gloves provide.
 Screw caps or caps that fit over the rim outside the centrifuge tube are safer than
Latex + Alcohol = Holes
plug-in closures. Some fluids usually collect between a plug-in closure and the rim
of the tube.
 Centrifugation may be performed in a centrifuge that is contained
within a specially designed biological safety cabinet or other
physical containment device. (Not available at CCM)
 If a whole centrifuge containment device is not available, internal
aerosol containment devices (e.g., sealed canisters, safety cups or
buckets with covers, heat sealed tubes or sealed rotors, etc)
should be used. Routinely available at CCM.
 Check sealed canisters before each use for cracks and verify the
proper placement of the black rubber “O”-ring. Apply vacuum
Sealed containment canister;
grease to O-rings if they look dry to prevent the rubber from
open in BSC
cracking and breaking (replace if cracked).
o Canisters are instrumental in containing liquid released from a cracked or broken tube.
Relatively few organisms are released into the air from a cracked tube contained in a
canister compared to breakage that releases the fluid into the centrifuge chamber (when
no canister is used).
 Infectious agent samples must be loaded into an aerosol containment device and closed within a
BSC and the aerosol containment device must be removed from the centrifuge and opened in a
BSC. If a BSC is unavailable, a minimum of 10 minutes settling time should be allowed before
opening.
Safe work practices for blending, sonicating, grinding, and lyophilizing of biohazards



Operate blender, sonicator, and grinder in a BSC, or place a towel moistened with disinfectant
over the top of blender, grinder, or sonicator.
Use safety blenders designed to prevent leakage. Blenders are potent aerosolizers and their use
must be carefully monitored. A written safety protocol must be included in your BUA prior to the
start of work. Alternative methods are available to avoid the use of blenders.
Filter lyophilizer vacuum pump exhaust through HEPA filters (e.g. Whatman Vacu-Guard filter
6722-5001) or vent into a BSC.
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
Autoclave or disinfect all equipment promptly after use.
Safe work practices for pipetting of biohazards
Aerosols are created during pipetting procedures by liquid dropping from a pipette to a work
surface, by mixing cultures by alternate suction and blowing, by forceful ejection of an
inoculum onto a culture dish, or by blowing out the last drop. It has been demonstrated by
high-speed photography that an aerosol of approx 15,000 droplets is produced when the last
drop of fluid in the tip of the pipette is blown out with moderate force.



Pipette biohazardous materials in a BSC, if possible.
Drain a pipette with tip against the inner wall of the receiving vessel. Avoid forcibly
expelling hazardous material from a pipette.
Discharge from pipettes should be as close as possible to the fluid or agar level, not
dropped from a height.
Mouth pipetting is prohibited; always use a mechanical pipetting device.
Avoid bubbling expiratory air through a biohazardous liquid with a pipet.
Culture Plates, Tubes and Bottles


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
Use a shielded electric incinerator or hot bead sterilizer to sterilize inoculating loops.
Disposable plastic loops and culture needles are good alternatives to open flames.
NOTE: New laboratory safety requirements (7/13) may require use of Flame
Retardant lab coats to use flammable liquids (ethanol) and open flames.
The insertion of a sterile, hot wire loop or needle into a liquid or slant culture can
cause spattering and release of an aerosol. To minimize the aerosol production,
cool the loop in the air or by touching it to the inside of the container or surface of the
agar prior to contact with the culture or colony.
Water in Petri dish cultures usually contain viable microorganisms and form a film
between the rim and lip of the inverted plate. Aerosols are dispersed when this film is
Ceramic Loop
broken by opening the plate. Vented plastic Petri dishes where the lid touches the
Incinerator
rim at only three points are less likely to offer this hazard. The risk may also be
minimized by using properly dried plates.
If the sediment in the tube or bottle is packed, infectious microorganisms must be carefully resuspended to minimize the amount of aerosol created. It is better to use a swirling, rotary motion
rather than shaking. Shaking always contaminates the closure; thus, there is
the added hazard of liquids dropping from the closure or running down the
outside of the container.
20

Mechanical
Pipetting Device
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15


Touch tip to side
of tube
Syringes and Needles
 The hypodermic needle is a dangerous instrument – never forget!


Vortexer
Of all the accidents at CCM, the ones with the most dangerous and disturbing consequences
have involved needles with concentrated infectious agents.
Always, always, always handle hypodermic needles with infectious agents at the highest level of
respect
o TAKE YOUR TIME - do not be in a hurry
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MANAGE YOUR TIME REALISTICALLY - do not try to do too much
SPEAK UP if you feel uncomfortable with the work load and the time
frame; break it up into smaller, more manageable, parts; take rest
breaks
o DON’T LET DOWN YOUR GUARD - most accidents happen when
you become comfortable with the tasks
o DOCUMENT A DRY RUN – perform the procedure first with water or
PBS and document your dry run as part of your protocol.
Needle stick
When infectious agents are involved, use the syringe and needle in a BSC and
avoid quick and unnecessary movements of the hand holding the syringe. Clear the BSC of
unnecessary material and equipment in preparation for loading infectious agents into syringes
with hypodermic needles.
Use needle-locking (Luer-Lok type) syringes only and be sure that the needle is locked securely
into the barrel. A disposable syringe-needle unit (where the needle is an integral part of the unit)
maybe preferred for most procedures. Safety devices are strongly encouraged!
Try wearing surgical or other type rubber gloves for all manipulations with needles and syringes
to increase protection from accidental autoinoculation.
Fill the syringe carefully to minimize air bubbles and frothing of the inoculum.
Expel excess air, liquid and bubbles from a syringe vertically into a cotton pledget moistened with
the proper disinfectant or into a small bottle of sterile cotton.
Do not use the syringe to expel forcefully a stream of infectious fluid into an open vial or tube for
the purpose of mixing. Mixing with a syringe is condoned only if the tip of the needle is held
below the surface of the fluid in the tube.
When removing a syringe and needle from a rubber-stoppered bottle, wrap the needle and
stopper in a cotton pledget moistened with the proper disinfectant.
Inoculate animals with the hand “behind” the needle to avoid punctures.
Be sure the animal is properly restrained prior to the inoculation and be on
the alert for any unexpected movements of the animal.
Before and after injection of an animal, swab the site of injection with a
disinfectant.
Safety Needle with
Discard contaminated syringes into a sharps container WITHOUT removing
Sliding Cover
the needle or manually replacing the protective needle sheath that is
furnished with disposable hypodermic needles or syringe-needle units.
Use safety needles whenever possible; annually review available safety needles as new products
are frequently becoming available.
Where the protective needle sheath must be replaced following use of a syringe, forceps should
always be used to minimize the possibility of exposure via accidental autoinoculation. Single
hand technique ONLY.


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
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15
o
o
Other safety precautions




Wear gloves when handling infectious materials, or infected animals
If a spill occurs that may generate aerosols, stop breathing until you leave the area (without
taking in a BIG breath), close the door, add signage to door to prevent entry, wait 30-60 minutes
to allow dissipation of aerosols before returning to clean up. See Exposure Control Plan
SOP#4: Guidelines for Biological Spill Clean Up (SafetyNet #27) for spill cleanup.
See UCD Environmental Health and Safety SafetyNet#21 Minimizing Aerosol Exposure
Report any spill of recombinant DNA or infectious agent as soon as possible to the Biosafety
Office. You may report online:
http://safetyservices.ucdavis.edu/ps/bis/biohazardIncidentReport

Report all injuries and accidental autoinoculation, ingestion or inhalation of infectious agents to
the laboratory director or supervisor, EH&S (call 530-752-1493) and the UC Davis Occupational
Health physician (530-752-6051) for evaluation and possible treatment. See SafetyNet #52:
Emergency Medical Care.
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Proper Use of Vacuum in the Laboratory
The use of vacuum to collect supernatant from samples is common in
laboratories, especially where tissue culture procedures are routinely used.
Vacuum systems improve the efficiency of many routine tasks. However, safety
procedures must be in place to protect users, co-workers, the building vacuum
system and the personnel who repair and maintain the system from
contamination with biohazardous materials or chemicals.
At CCM the main use of vacuum is to aspirate cell culture supernatants and chemical
supernatants during molecular biology procedures.
15
Where does the air go? If you were to follow the vacuum line pipes from your
laboratory bench to their beginning, then you would snake through the walls, around
and over the hallways and end up in the vacuum pump in the mechanical room
(1051) where the air is released back into the environment.
CCM Building Vacuum
Pump
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20
Proper Vacuum Set-up:
Fe
What could go wrong?
Safety Issues, Consequences and PROPER PROCEDURES for Vacuum Systems:
1. Aerosols: Drawing a vacuum may result in the aerosolization of microorganisms from within the
collection flask.
 Consequence: Contamination of the vacuum line, pump, the environment and the
exposure of repair and maintenance personnel who service the vacuum source.
 PROPER PROCEDURE: Include a HEPA filter designed to fit in the tubing between
your overflow flask and the vacuum outlet. Examples: Whatman HEPA-Vent filter
(Fisher cat# 09-744-79) or Whatman HEPA-Cap Disposable Air Filtration Capsule
(Fisher cat#09-744-15)
2. Liquid Contamination: Overflow of collection flask or absence of collection flask; liquid enters
at the same level as the vacuum exit (sucked out before it reaches the bottom!)
 Consequence: Can result is gross contamination of the vacuum lines with biohazards and
chemicals; cause malfunction of the building vacuum pump; in addition to the generation
of aerosols.
 PROPER PROCEDURE: Include a hydrophobic filter between your overflow flask and
your HEPA filter (prevent the HEPA from wetting). The liquid will collect only as far as
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5.
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4.
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3.
the hydrophobic filter, protecting the lines, and prevent further use of the vacuum until
the filter is replaced. To prevent both fluid and aerosol contaminations use a Whatman
Vacu-Guard filter (Fisher Cat# 09-744-76).
Implosions: Non-vacuum safe glassware or plastic ware can collapse.
 Consequence: Release sharp fragments, aerosols and fluid violently and
unexpectedly; contamination of personnel and equipment; time lost in
clean up procedures.
 PROPER PROCEDURE: Use ONLY glassware and plastic ware
specially designed for use under pressure. CCM sidearm Erlenmeyer
flasks are available for chemical and BSL1/2 applications; although
pressure approved plastic ware is always preferred to glassware. Example: Nalgene
Heavy-Duty Polypropylene Vacuum Bottles, Fisher Cat#06-443B. Review Attachment
#1: Implosion Accident. Shared laboratory spaces (1052, 2052, 2043A) and BSL3
(2042A/B) applications absolutely require the use of plastic ware and inline filters (see #1
and #2 above) for all vacuum applications.
Sharps: Glass (Pasteur) pipettes are sharps.
 Consequence: Autoinoculation possible with contaminated sharp or broken
glass Pasteur pipettes (discard in approved sharps containers).
 PROPER PROCEDURE: Avoid glass Pasteur pipettes when possible. Use
disposable aspiration pipettes in place of glass (Pasteur) pipettes (e.g., Fisher
cat#13-675-16) or use large non-filtered pipette tips or unplugged plastic
pipettes.
Sharps
Vacuum Line: Cracked and brittle vacuum line due to age and/or exposure to
Container
ultraviolet light.
 Consequence: Poor vacuum and possible leakage of liquid contents.
 PROPER PROCEDURE: Check vacuum lines for cracks; do not store
vacuum set-ups in a biological safety cabinet (BSC) where ultraviolet
light (UV) is used, as UV light degrades tubing.
Breakage or Spillage: Unprotected glass or improperly closed vacuum
collection flasks in an area where it can be easily pushed off the counter, kicked
or knocked by a rolling chair.
 Consequence: Exposure of users and co-workers to collection flask
contents and time lost in clean up procedures.
 PROPER PROCEDURE: Use plastic ware whenever possible to prevent
breakage; place collection flask in a secondary container if located on the Accident waiting to happen:
floor. Protect the container from bumps and knocks – place it out of
Unprotected flask under a
traffic areas and away from your feet and rolling chairs. Keep tubing out
BSC and exposed tubing
of harms way also!
Biohazards: Vacuum system is used to collect biohazardous liquids (i.e., virus culture
supernatants, human cells).
 Consequence: Exposure of users and co-workers to infectious substances due to an
accident from any of the issues listed above.
 PROPER PROCEDURE: All users and co-workers wear
appropriate personal protective equipment (PPE; gloves, lab coats
and safety glasses) while in the laboratory. Add disinfectant to the
vacuum collection container BEFORE collection of biohazardous
liquids and empty it promptly after disinfection is complete.
CAUTION: Dilute bleach is active for 24 hours! Make a fresh
solution at least once a day.
7.
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8. Unlabelled Contents: Unlabelled/unmarked collection flasks.
 Consequence: Chemical/Biohazardous waste must be properly labeled to avoid fines
(unlabelled chemical container); in the case of breakage or spillage – unknown contents
makes clean-up difficult, expensive and time-consuming.
 PROPER PROCEDURE:
o Label biohazardous collection flasks “inactivated bleach
product”. Add bleach (or appropriate disinfectant for the
biohazard) to the collection flask BEFORE collecting the
biohazardous solution. Discard promptly after disinfection
complete.
o Chemical collection is trickier, especially if the contents
must be collected for disposal by EH&S (see “NOTE”
below). Often the collection bottle for a vacuum system is
NOT the final collection bottle used for chemical waste
disposal – the contents are moved to a properly labeled
chemical waste bottle for pick-up. So, how do you label a
chemical collection bottle on a vacuum system without
Have you seen this nasty
incurring fines?
collection flask at CCM?
 First, do NOT label it “WASTE” in big letters –
especially if it sits for weeks on the bench top!
 Second, remember that it is not “waste” until you determine it is – as
long as your procedure is part of your WRITTEN laboratory protocol –
consider it a “holding” container until you are sure you don’t “need” it
for any other reason, and then move it to a properly labeled UCD waste
container for pick-up by EH&S.
 Label the flask with what is in it – “Dilute Sodium Hydroxide Solution”
or “Neutralized Qiagen Plasmid Prep Supernatant” or…you get the idea.
o Dilute Buffer solutions: often the collection consists of very low concentration
buffers that may be discarded down the sink (e.g. phosphate buffered saline).
Label the collection flask as “Used PBS buffer” (for example).
 NOTE: the campus has a ZERO tolerance for solutions containing
COPPER– no discharge is allowed down the sink – copper must be
collected for disposal by EH&S. See SafetyNet#6: Can this go down
the drain? Many immunological assays use copper.
Good laboratory practices: Get in the habit – create the label at the same time as the solution. Label the
solution on the stir plate and move the label to the final solution container. You have to make a label –do
it sooner, rather than later!
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ccm --bio,chem and animal - CCM Safety Training

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