Biological Safety Training

1
Biological Safety Training

• Radiological and Environmental Management
• Environmental Health
• Purdue University
• October 2003

2
Institutional Biosafety Committee (IBC) Procedures

• The Universitys Institutional Biosafety
  Committee (IBC) is the campus-based committee
  that has the responsibility for reviewing and
  approving all proposals, activities, and
  experiments involving recombinant DNA,
  biohazardous materials, and unfixed Human
  tissues, cell lines, or fluids. The IBC reviews
  processed protocol applications that deal with
  Class II or higher biohazards, unfixed human
  blood or tissues, or recombinant DNA materials.
  Principal Investigators must complete an IBC
  protocol application (Form 1A).

3
Institutional Biosafety Committee (IBC)
Procedurescontinued

• Form 1A covers the basic project information, a
  facility inspection, a listing of personnel who
  will be involved, and assures that bio-awareness
  training is provided.
• The Biosafety Officer then conducts a lab or
  facility inspection and assigns a biosafety
  level. The IBC Chair completes the process by
  reviewing the project procedure and either
  approves, disapproves, or exempts the protocol.
• Go to IBComm_at_purdue.edu to contact the
  committee.

4
NIH Biohazard Risk Groups

• Class 1 Agents of no or minimal hazard under
  ordinary conditions or handling.
• Class 2 Includes agents which may produce
  disease of varying degrees of severity from
  accidental inoculation or injection or other
  means of cutaneous penetration but which are
  contained by ordinary laboratory techniques.
• Class 3 Includes pathogens that require special
  conditions for containment.
• Class 4 Require the most stringent conditions
  for their containment because they are extremely
  hazardous to laboratory personnel or may cause
  serious epidemic disease.

5
Biohazard Exposure

• BIOHAZARDS
• Any organism/biologic by-product/allergen that
  causes ill effects to people, plants, or animals.
• Can be either a fungus, bacteria, virus, prion,
  parasite, toxin, or DNA component.
• Can be human body fluids, unfixed tissues, cell
  lines, or genetic oncogenes.
• EXPOSURE to biohazards may occur from
• Puncture wounds
• Contact with abraded skin
• Absorption through respiratory tract, oral route,
  or mucous membranes.
• Principal Investigators who are unsure if a
  material is a biohazard should contact the
  University Biosafety Officer, ibcomm_at_purdue.edu

6
Biohazard Classifications

• Biosafety levels corresponds to the Class rating
  of the biohazard, i.e., BSL2 is required for
  Class II biohazards. BSL2 is the most common
  type of biohazard project that occurs on campus.
  Labs working under these conditions must have the
  following procedures in place
• Limited access,
• Door posts warnings,
• Surface decontamination methods,
• Waste handling,
• No eating or drinking,
• Hand washing,
• Appropriate personal protective equipment, and
• A Class 2 Biosafety Cabinet (if aerosol
  production is anticipated).

7
Bio-Awareness Training

• Specific handling techniques for each class II or
  higher pathogen that is used must be reviewed and
  explained by the Principal Investigator.
• Signs and symptoms for the onset of disease
  caused by the organism must be reviewed.
• Special hazards, i.e., complications for
  pregnancies and immuno-compromised lab staff,
  must be explained and posted.
• Appropriate decontamination procedures must be
  reviewed.
• Any required security steps must be taken.

8
Personal Protective Equipment (PPE)

• PPE
• The last barrier of defense between the user and
  the hazard.
• Must be available for every body part as well as
  respiratory protection.
• Proper selection is critical for each individual.
• Know how to don and doff each piece of PPE.
• A typical selection of PPE for biohazard
  application
• would be the use of nitrile or latex gloves,
  goggles, lab
• coat, closed toe shoes, and perhaps a N-95 dust
  and
• vapor mask.

9

• PPE can include the PROPER
• Goggles
• Gloves
• Footwear
• Lab Coats
• For the Task

10
Dust and Vapor Masks

• Dust and vapor mask such as N-95 and N-100, are
  designed to trap minute particles and minor
  aerosols. These are recommended for use when
  handling biosafety level 2 agents outside of a
  biosafety cabinet or if there are aerosols
  produced in environments such as animal BSL2
  facilities.
• There are many different styles of dust and vapor
  masks available making it important to find a
  mask that will provide optimum facial fit and
  comfort.

11
Respiratory Protection comes in many varieties.
Choose the appropriate mask for the task.
12
Waste Handling - Biohazard Bags

• BIO-RELATED WASTE MATERIALS
• Dispose of in properly labeled water impervious
  autoclavable bags (usually red or orange bags).
• Do not over fill bags.
• Utilize proper personal protective equipment and
  wash hands after completion.
• Keep bags closed until they are ready to be
  decontaminated.
• Do not dispose of a bio waste bag in the regular
  trash.
• Do not place these bags in public areas such as
  hallways. Contact REM for removal of deconned and
  autoclaved materials.

13
Waste Handling - Biohazard Bagscontinued
ALWAYS REMEMBER When handling a bio waste bag,
be careful not to press the bag against your
body. This prevents improperly disposed of sharp
accidents. Do not create aerosols by forcing the
air out of the waste bag.
14
Protect yourself from the contents. Hold Bio bags
away from your body. Dont squeeze the bag so
that aerosols are released into your breathing
space. Take biowaste to the autoclave room as
soon as possible.
15
Waste Handling - Autoclave

• Autoclaves are very effective decontamination
  devices that use high pressure and steam (270?F)
  to eliminate biological activity on equipment,
  wastes, or growth mediums.
• Extreme heat and high pressure produced in
  autoclaves can cause serious injury. Always read
  the operations manual and review usage procedures
  with the lab manager. Users should understand and
  respect the hazards these can create and use the
  proper personal protection equipment, i.e.,
  heat-resistant gloves, lab coat, and face shield.
  
• Do not put the following items in the autoclave
• - Sealed containers - Aerosol cans
• - Volatiles - Uncontained
  sharps

16
Waste Handling - SHARPS

• SHARPS are items such as glass and plastic
  pipettes, broken glass, test tubes, petri dishes,
  razor blades, needles, syringes, etc. that are
  capable of puncturing, cutting, or abrading the
  skin.
• Properly handle, decontaminate, and dispose of
  sharps in order to prevent injury and potential
  disease transmission.
• Dispose of in properly labeled puncture proof
  containers and never discard in the regular
  trash.
• Call REM for pickup.

17
DO NOT PLACE SHARPS CONTAINERS IN THE REGULAR
TRASH!
18
ALWAYS place sharps in puncture resistant
containers for disposal.
19
Waste Handling SHARPScontinued
BE CONSIDERATE Laboratory employees that
routinely work with sharps and building services
personnel who handle wastes are at risk of being
punctured or lacerated during their
workday. Report sharp related injuries to your
supervisor and to the Biosafety Officer
immediately.
20
Biological Safety Cabinets
Laminar Flow Clean Benches

• Protects operator
• Protects the environment
• Protects the product
• Reduces the risk of airborne infection by
  reducing the escape of aerosols

• Protects the product
• Blows aerosols into the operators face.
• Does not protect the operator
• Should not be used with potential pathogens
• Is not a Biological Safety Cabinet

21
Biosafety Cabinet Procedures

• Biological safety cabinet usage must be combined
  with good work practices for optimum safety and
  contamination control.
• Movement of arms into and out of the cabinet can
  disrupt airflow, adversely affecting cabinet
  performance. Avoid bringing non-essential
  equipment and supplies into the cabinet.
• Place supplies, equipment and absorbent towels so
  that air intake or exhaust grilles are not
  obstructed.

22
Biosafety Cabinet Procedurescontinued

• Keep opening and closing of lab doors and other
  personnel activity to a minimum.
• If a burner is deemed to be indispensable use an
  on demand type.
• Work at least 4-6 inches inside the cabinet
  window.
• Decontaminate spills as soon as they occur
  remove and disinfect the grille if contaminated.
• For more instructions see Purdues Biological
  Safety Manual

23
Biohazard Spills

• Evacuate the lab for thirty minutes if the spill
  is a large volume of biohazard material. This
  allows aerosols to settle.
• Lab staff decontaminating small biohazard spills
  need to have specific bio-agent awareness
  training, and plan in advance for cleanup of
  emergency spills.
• If spill occurs inside a Biosafety Cabinet, leave
  cabinet turned on and decontaminate in place.
• Use the proper personal protective equipment
  (PPE), e.g. latex gloves, outerwear, goggles, etc.

24
Biohazard Spillscontinued

• Spray the contaminated surfaces with a 1-10
  solution of bleach and water. Be careful not to
  contaminate the outside of the spray bottle.
• Remove all traces of the spill with paper towels
  or other acceptable materials and re-spray the
  cleaned area with the bleach solution and allow
  the air to dry.
• Place all waste materials, including disposable
  PPE, into an autoclavable biohazard bag. Be
  careful not to contaminate the outside of the
  bag.
• REPORT ALL SPILLS TO YOUR SUPERVISOR AND THE
  BIOSAFETY OFFICER

25
In the event of a spill Use the proper PPE Use
the proper disinfectant solution. Dispose of
clean-up materials in biohazard bags.
26
COMMONLY USED DISINFECTANTS

• Alcohols ethyl or isopropyl alcohol at 70-80
  percent concentration is a good general purpose
  disinfectant not effective against bacterial
  spores.
• Phenols Effective against gram-positive
  bacteria, fungi, and lipid-containing viruses.
• Quaternary Ammonium Compounds Cationic
  detergents which are strongly surface active
  extremely effective against lipophilic viruses
  not effective against bacterial spores.
• Chlorine Low concentration (50-500 ppm) active
  against vegetative bacteria and most viruses
  higher concentration (2500 ppm) required for
  bacterial spores corrosive to metal surfaces
  must be prepared fresh laundry bleach (5.25
  percent chlorine) may be diluted and used as a
  disinfectant.
• Iodine Recommended for general use effective
  against vegetative bacteria and viruses poor
  activity against bacterial spores. 
• Many of these disinfecting agents can be
  irritating and toxic and should be used in
  accordance with label direction for personal
  protective equipment, concentration, and contact
  time.

27
Security

• EVERYONE is responsible for security in the
  laboratory.
• If you are using Class II biohazards make sure
  these materials are secured.
• Keep your lab doors locked as well as the doors
  to areas outside your lab where Class II
  organisms are incubated, stored, or otherwise
  processed.
• Ask for identification from unknown individuals
  who enter your lab.
• Do not allow access to unauthorized people.
• Secure biohazardous waste and autoclave as soon
  as possible.
• Report the disappearance of any biohazardous
  material to your Department Head and to the
  University Biosafety Officer.
• Special security measures apply to users of
  Select Agents. See the Biosafety Officer for
  more specific details.

28
Resources

• Center For Disease Control and Prevention
•  http//www.cdc.gov/
•  Select Agent Program
•  http//www.cdc.gov/od/sap/
•  Biosafety in Microbiological and Biomedical
  Laboratories 4th Edition, May 1999
•  http//bmbl.od.nih.gov/
•  Guidelines for Research Involving Recombinant
  DNA Molecules (NIH Guidelines)
• http//www.niehs.nih.gov/odhsh/biosafe/nih/rdna-ap
  r09.pdf  

29
Resourcescontinued

• Material Safety Data Sheets For Research
  Pathogens
• http//www.hc-sc.gc.ca/hpb/lcdc/biosafty/msds/inde
  x.html 
• Purdue University Biosafety Manual
•  http//www.adpc.purdue.edu/PhysFac/rem/home/bookl
  ets/bioman.htm

30
Quiz

• Please take the following review quiz, then check
  your answers on the last slide.
•  1)  Why is it important for lab staff to have
  bio-
• agent awareness training?
• a)  Some agents can cause serious complications
  for pregnancies and immuno-compromised
  individuals.
• b)  Signs and symptoms can determine if an
  exposure has taken place.
• c)  De-con procedures can be determined at that
  time.
• d)  All of the above.

31
2)  While working with an organism that has an
exposure route including skin contact and
respiratory, what personal protective equipment
(PPE) is needed? a)  Biosafety cabinet b) 
Respiratory or N-95 mask c)  Protective outer
wear, gloves and goggles d)  All of the
above 3) Why is posting a Hazard Assessment
important ? a)  It is an accessible information
source for employees to use to determine proper
personal protective equipment. b)  It is
required by the Center For Disease
Control. c)  It is an ineffective bureaucratic
hoop to jump.
32
4) What is inappropriate laboratory personal
protective equipment? a)  Gloves b)  Open toed
shoes c)  Goggles 5) Sharps are defined as
anything that will cut, puncture, or abrade the
skin. True False 6) What should be done in
the event of a bio-exposure sharps
injury? a)  Wash the site with soap and
water b)  Inform your supervisor c)  Contact
REM d) All of the above
33

• 7) Biohazard bags should not be.
• a) overfilled.
• b) disposed of in the regular trash.
• c) held against your body or squeezed
• d) All of the above
• 8) If a spill occurs inside of a Biological
  Safety Cabinet, which of the following statements
  is not appropriate procedure to follow?
• a) Report the spill to your supervisor
  immediately.
• b) Clean it up fast, before anyone sees it.
• c) Wear the proper Personal Protective Equipment
  for the material for which you are working.
• d) Plan in advance for an emergency. For
  example, what supplies and equipment should you
  maintain in your area to assist you in the event
  of an accidental spill?

34
9) Who is responsible for security in a BSL2 or
higher level laboratory? a) Purdue Police b)
The supervisor c) The Building Deputy d)
Everyone is responsible for security in the
laboratory. 10) Who should be contacted for
questions about Biosafety related issues? a)
University Police b) Biosafety Officer c)
Building Deputy d) Radiation Safety Officer
35
QUIZ Answer Key

• 1) D 8) B
• 2) D 9) D
• 3) A 10) B
• 4) B
• 5) T
• 6) D
• 7) D