Biological Safety Manual - Environmental Health & Radiation Safety

94 Pages·2013·4.24 MB·English

BIOLOGICAL
SAFETY
MANUAL
Version 1.0
2013

Environmental Health & Radiation Safety
3160 Chestnut Street, Suite 400
Philadelphia, PA 19104-6287
215-898-4453
[email protected]
TABLE OF CONTENTS
SECTION Page
EMERGENCY CONTACTS ................................................................................ 4
INTRODUCTION ................................................................................................. 5
FREQUENTLY ASKED QUESTIONS ................................................................. 8
BIOLOGICAL RISK ASSESSMENT .................................................................. 10
BIOSAFETY LEVELS ........................................................................................ 14
ANIMAL BIOSAFETY LEVELS ......................................................................... 20
BIOSAFETY CABINETS ................................................................................... 25
COMMON LAB EQUIPMENT ........................................................................... 30
INFECTIOUS AGENTS ..................................................................................... 34
RECOMBINANT DNA ....................................................................................... 36
HUMAN SOURCE MATERIAL .......................................................................... 38
NON-HUMAN PRIMATE MATERIAL ................................................................ 41
SELECT AGENTS ............................................................................................. 44
BIOLOGICAL RESEARCH LABORATORY ...................................................... 46
DECONTAMINATION ....................................................................................... 51
INFECTIOUS WASTE MANAGEMENT ............................................................ 54
SHARPS WASTE MANAGEMENT ................................................................... 58
MIXED WASTE MANAGEMENT ....................................................................... 60
ANIMAL INFECTIOUS WASTE ........................................................................ 61
AUTOCLAVING INFECTIOUS WASTE ............................................................ 62
TRANSPORT & SHIPPING OF BIOHAZARDS ................................................ 65
APPENDIX A – BIOLOGICAL AGENTS ........................................................... 67
RISK GROUPS: Bacteria .............................................................................. 67
RISK GROUPS: Fungi .................................................................................. 72
RISK GROUPS: Viruses ............................................................................... 73
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TABLE OF CONTENTS
RISK GROUPS: Parasites ............................................................................. 82
APPENDIX B – BIOSAFETY CABINET AIR FLOW .......................................... 85
APPENDIX C – SELECT AGENTS & TOXINS ................................................. 92
RESOURCES ................................................................................................... 93

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EMERGENCY CONTACTS

IF YOU ARE INJURED AND REQUIRE ASSISTANCE ON CAMPUS:
511
From a CAMPUS PHONE Call:
215‐573‐3333
From Your PERSONAL PHONE Call:

For medical assistance
DURING WORK HOURS (Mon – Fri 8AM - 4:30PM)

FACULTY AND STAFF report to:

HUP OCCUPATIONAL MEDICINE
Penn Tower, 4th Floor
300 South 33rd Street
215-662-2354

STUDENTS report to:

STUDENT HEALTH SERVICE
3535 Market Street, Suite 100
215-746-3535

For medical assistance AFTER WORK HOURS and HOLIDAYS:

ALL report to:

HUP EMERGENCY DEPARTMENT
Ground Floor Silverstein Pavilion
34th & Civic Center Blvd.
215-662-3920

ENVIRONMENTAL HEALTH AND RADIATION SAFETY

215-898-4453

Version 1.0 2013 Page 4
SECTION 1.0 INTRODUCTION

PURPOSE (PIs), laboratory supervisors, or laboratory
managers must contact the Office of

Environmental Health & Radiation Safety by
The purpose of the Biosafety Program is to
phone [phone]) or email
protect all employees, students, the public, and
([email protected]), if they are uncertain
the environment from exposure to biological
how to categorize, handle, store, treat or discard
agents or materials being used at the University
any biologically derived material.
that may cause disease or be harmful to

humans. This manual provides a

comprehensive overview of proper work
ROLES AND RESPONSIBILITIES
practices, regulations, and requirements for
proper containment and disposal of biological
hazards.
Office of the Vice Provost for

Research
POLICY
The Vice Provost for Research has responsibility
for the development and implementation of
research policies and procedures across the
The University of Pennsylvania is committed to
University.
providing a healthy and safe learning, teaching,
research, and work environment. Accordingly, The Vice Provost chairs the Provost's Council on
the goals of the University's Biological Safety Research which has representatives from the 12
Program are to: component schools of the University and
advises the Vice Provost for Research on
 Ensure a HEALTHY and SAFE research
formation and implementation of research
environment.
policies.
 PROTECT staff, students, and community The Vice Provost is also responsible for
from exposure to infectious agents. administering, overseeing, and coordinating a
wide variety of activities. In order to provide
 PREVENT environmental contamination.
comprehensive services to researchers, the
Office of the Vice Provost for Research unites
 SECURE experimental materials.
and coordinates the following offices:
 COMPLY with Federal, State and Local
 Office of Research Services (ORS) works
Regulations.
with researchers, centers, schools, and
funders on the financial and contractual
aspects of sponsored projects.
The Office of Environmental Health & Radiation
Safety (EHRS), under the direction of the  Office of Regulatory Affairs (ORA) administers
University's Institutional Biosafety Committee
the University's program of compliance in the
(IBC) and The Office of the Vice Provost for areas of human subjects (IRB) and the care
Research, developed this Biological Safety and use of animals in research (IACUC).
Manual. This manual provides university-wide
safety guidelines for working with biological  University Laboratory Animal Resources
hazards (biohazards). It also outlines general (ULAR) handles care and husbandry of
policies and procedures for using and disposing laboratory animals.
of infectious or other potentially infectious
materials (OPIM). Penn biosafety policies  Office of Environmental Health and Radiation
ensure compliance with federal, state, and local Safety (EHRS) performs risk assessments,
laws, regulations and guidelines. This manual is provides awareness training, resources and
a guidance document that changes in response regulatory guidance to ensure safe conduct of
to changes in existing regulations and inclusion research. Oversees and reviews use of
of new regulations. It may not address all biological, chemical, and radiation hazards in
hazards encountered by faculty, students, staff the University's research programs.
and the Penn community.
 Center for Technology Transfer (CTT) obtains
Biological safety practices and procedures in all
and manages patents, copyrights, and
University laboratories must comply with those
trademarks derived from the University's
outlined in this manual. Principal investigators
Version 1.0 2013 Page 5
SECTION 1.0 INTRODUCTION
academic research enterprise. and personnel contamination.
In addition, the Office of the Vice Provost for  The Vice Provost for Research appoints
Research works closely with the Office of members of the IBC.
Human Research and serves as the University
of Pennsylvania's Research Integrity Officer.
The Office of Environmental Health &
Research Compliance and Radiation Safety (EHRS)
adherence to EHRS guidelines  The Office of Environmental Health and
Radiation Safety (EHRS) is the operational
for research at the University of
arm of the Institutional Biosafety Committee
Pennsylvania is mandated by the (IBC). It provides instruction and training on
safe work practices, conducts routine
Environmental Protection Policy
inspections of work areas, investigates
Statement (October 7, 2008). accidents and recommends
preventive/corrective actions, reviews animal
Compliance with University, research protocols involving hazardous
materials, reviews renovations and new
federal, state and local
construction design for safety features and
regulations is a condition of responds to emergencies.
acceptance of research funding  The Institutional Biosafety Officer (IBSO) is an
EHRS Associate Director and is responsible
from the NIH and various other
for oversight and daily implementation of the
granting agencies. Biosafety Program.
 The IBSO is responsible for compliance with
all federal, state, and local regulations that

apply to biosafety and keep University
The Institutional Biosafety Committee regulations up to date.
(IBC)
 Biosafety Officers (BSO) are EHRS staff
 The Institutional Biosafety Committee (IBC) members and reports to the Institutional
has University-wide oversight as mandated by Biosafety Officer (IBSO). BSOs perform risk
the National Institutes of Health (NIH) Office of assessments on labs as needed, help PIs
Biotechnology Activities (OBA) and is charged develop SOPs, offer appropriate training to all
by the Vice Provost for Research with staff and students, and perform annual
formulating policy and procedures related to laboratory audits of research spaces where
the use of biohazardous agents, including: biological materials are used.
human pathogens, oncogenic viruses, other
 BSOs review work requiring BSL-3 or ABSL-3
infectious agents, human gene transfer, and
containment, pre-review rDNA registrations,
recombinant DNA.
maintain the Biological Agent Registration
 The IBC is responsible for review and (BAR) and administer the Select Agent
approval of projects involving recombinant Program at Penn.
DNA research and human gene transfer
 BSOs will respond to and follow up on any
protocols. Additionally, the IBC reviews work
major biological incident and/or spill as
with Select Agents, biohazardous agents that
needed. BSOs should be called to assist with
are animal or human pathogens requiring
large spills containing infectious material.
BSL-3 or ABSL-3 containment, oncogenic
They will assist the PIs to ensure that all
viruses, and other potentially infectious agents
corrective actions and emergency procedures
on an as needed basis.
are followed in accordance with applicable
 The committee sets containment levels in University regulations and guidelines.
accordance with National Institutes of Health
(NIH) and Centers for Disease Control and
Prevention (CDC) guidelines, and adopts
emergency plans covering accidental spills
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SECTION 1.0 INTRODUCTION
Deans/Department Chairs
 PIs are responsible for hands on training for all
 Deans/Department Chairs are responsible for laboratory procedures. They must ensure that
the implementation of safe practices and all laboratory staff has fulfilled University
procedures in their schools or departments. training requirements and are current in all
required training.

 The PI and/or lab personnel are responsible
Facilities Departments
for initiating cleanup and disinfection in the
event of a biohazard spill in a laboratory. If
 The Facilities Department in each school is
assistance is required contact EHRS. Once
responsible for the removal, packaging, and
the material has been contained, absorbed,
shipment of all infectious waste in accordance
and removed, housekeeping/facilities
with local, state, and federal regulations.
management should be contacted to finalize
the cleanup and disinfection of the area. The
 Building Administrators are responsible to
PI is responsible for ensuring that all
keep common laboratory spaces clean and in
corrective actions and emergency procedures
safe working condition. They are responsible
are followed in accordance with applicable
for getting broken infrastructure in the
University procedures and regulations.
laboratory fixed.

Principal Investigators (PIs) Employees and Students
 Laboratory staff and students are ultimately
 The Principal Investigator (PI) is responsible
responsible to follow the PI’s instructions for
for full compliance with approved research
working in the laboratory.
protocols, trainings required by the University,
the University Biological Safety Manual, the  All personnel working in Penn laboratories
NIH Guidelines (NIH Guidelines for Research
with potentially infectious materials must be
Involving Recombinant DNA Molecules), the
familiar with University training requirements
Occupational Safety and Health Administration
and the University Biological Safety Manual.
(OSHA) Bloodborne Pathogen Standard
Additionally, laboratory staff and students
(human-derived materials) and other local,
must be familiar with the approved research
state and federal regulations that apply to
protocols, the NIH Recombinant DNA
research.
Guidelines, and the Occupational Safety and
Health Administration (OSHA) Bloodborne
 In the laboratory, PIs must conduct a risk
Pathogen Standard.
assessment to identify potentially hazardous
procedures involving infectious agents,  Laboratory staff and students must understand
develop Standard Operating Procedures
how to safely work with potentially infectious
(SOPs), instruct and train all staff and students
agents, be provided and wear appropriate
working in the lab on safe work practices,
personal protective equipment (PPE), keep
keep the lab space clean and up-to-date, and
their laboratory space clean and up-to-date,
follow regulations for disposal of infectious
and follow regulations regarding the disposal
waste. The PI must provide PPE to their staff.
of infectious waste.
 PIs must register research projects that  All employees must receive proper training for
require review by the IBC and/or EHRS, such
their specific tasks, including hands on training
as the generation and/or use of recombinant
for laboratory procedures. They must also be
DNA, work requiring BSL-3 or ABSL-3
current in University training requirements.
containment, Select Agents, and other work
This requirement relies upon mandatory,
with infectious agents as needed.
annual completion of Penn Profiler which will
result in assignment of appropriate training in
 PIs must complete the Biological Agent
Knowledge Link.
Registration (BAR) and update it annually or
as needed.
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SECTION 2.0 FREQUENTLY ASKED QUESTIONS

 Why does my door sign say Biosafety Level 2?
Biosafety Levels (Section 3.2)
 How does a Biosafety Cabinet work?
Biosafety Cabinets (section 4.1)
 How do I safely work with my lab equipment?
Common Lab Equipment (section 4.2)
 What is recombinant DNA?
rDNA (section 5.2)
 What should I know when working with human material (blood, tissue, cells, etc.)?
Human Source Material (section 5.3)
 How do I work safety with non-human primate materials?
Non-human Primate Material (section 5.4)
 I want to work with a Select Agent.
Select Agents (section 5.5)
 What does a BSO look for during a lab audit?
Biological Research Laboratory – Annual Lab Audits (section 6)
 Help! My room sign is out of date.
Biological Research Laboratory – Room signs (section 6)
 I purchased a new centrifuge to spin down my virus supernatant. Does it need a special
label?
Biological Research Laboratory – Hazard Labels (section 6)
 When should I be wearing gloves?
Biological Research Laboratory – Hand Protection (section 6)
 I spilled my 50 ml tube of viral supernatant in the BSC. What do I do now?
Decontamination (section 7)
 My biohazard bag is full. What do I do with it?
Infectious Waste Management (section 8.1)
 How do I dispose of my used syringe?
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SECTION 2.0 FREQUENTLY ASKED QUESTIONS
Sharps Waste Management (section 8.2)
 Can I throw my used serological pipette in the biohazard bag?
Sharps Waste Management (section 8.2)
 What do I do with infectious waste that has been contaminated with a chemical?
Mixed Waste Management (section 8.3)
 Do I need to wear PPE while using the autoclave?
Autoclaving Infectious Waste (section 8.5)
 I’ve dissected a research mouse in my lab. Where do I dispose of it?
Research Animals Infectious Waste (section 8.4)
 Do I need training to ship a package with dry ice?
Transport & Shipping of Biohazards (section 9)
 Do I need a permit to ship my virus sample to my research friend in New Jersey (or anywhere
else)?
Transport & Shipping of Biohazards (section 9)
 What are animal biosafety levels (ABSL)?
Animal Biosafety Levels (section 3.3)
 How do I properly connect a vacuum line and vacuum flask?
Vacuum System Protection (section 4.1)
 Do I need a natural gas connected to a biosafety cabinet?
Alternatives to Continuous Flame Burners (section 4.1)

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SECTION 3.1 BIOLOGICAL RISK ASSESSMENT

RISK ASSESSMENT

On the basis of the information ascertained 10. Information available from animal studies
during the risk assessment, a Biosafety Level and reports of laboratory-acquired infections
can be assigned to the planned work, or clinical reports
appropriate personal protective equipment
selected, and standard operating procedures 11. Laboratory activity planned (sonication,
(SOPs) incorporating other safety interventions aerosolization, centrifugation, etc.)
developed to ensure the safest possible conduct
of the work. 12. Any genetic manipulation of the organism
that may extend the host range of the agent
or alter the agent’s sensitivity to known,
Conduct a risk assessment to determine the
effective treatment regimens (see Section
proper work practices and containment
5.2: recombinant DNA)
requirements for work with biohazardous

material. Risk assessments should identify

microorganism, their NIH established Risk
13. Experience and skill level of at-risk
Groups (see Table 3.1) and host/environmental
personnel, including safety training or
factors that influence potential exposure risks for
hands-on experience
workers. The following points can be used as a

guide but a Biosafety Officer should be
14. Local availability of effective prophylaxis or
consulted to ensure full compliance with
therapeutic interventions (immunizations or
established guidelines and current regulations.
post-exposure prophylaxis)

1. Pathogenicity or Risk Group of
15. Medical surveillance program
microorganisms

2. Infectious dose needed to cause infection in Bloodborne pathogen materials are
a healthy person
designated RG-2 and the BMBL

3. Potential outcome of exposure specifies BSL-2 containment

4. Natural route of infection (aerosol, ingestion, practices for bloodborne pathogen
skin contact)
materials in compliance with the

5. Other routes of infection, resulting from OSHA Bloodborne Pathogens
laboratory manipulations (parenteral,
airborne, ingestions); see Equipment Standard
Hazards below

6. Stability of the agent in the environment
IMPORTANT CONSIDERATIONS
7. Concentration of the agent and volume of WHEN PERFORMING A RISK
concentrated material to be manipulated
ASSESSMENT?
(tissue samples, blood, serum, etc.)

8. Origin of microorganism refers to geographic Identifying the agent or infectious
location, host (animal or human) or nature of material
source (potentially zoonotic, associated with

a disease outbreak)
Directors and principal investigators of

microbiological and biomedical laboratories have
9. Presence of a suitable host (human or
the important responsibility to do a risk
animal)
assessment of their laboratories in order to alert

their staff to the hazards of working with
infectious agents and to the need for developing
proficiency in the use of selected safe practices
and containment equipment to prevent
Laboratory Acquired Infections (LAIs).
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