Download Occupational Safety - College of Agriculture and Life Sciences

Biosafety in the Workplace
PLS 4/595D /Regulations and
Laboratory Management
Spring Semester, 2007
Mark J. Grushka, M.S., CSP
Manager, Biosafety and Biosecurity
University of Arizona
Main Topics
Part 1 Introduction to Biological Safety
Principles (Thursday/March 29th)
 Part 2 Introduction to Regulatory
Framework (Thursday/March 29th)
 Part 3 Biosafety Program Management,
Application of Project Management
Techniques and Case Studies
(Thursday/April 5th)

Part 1 Introduction to Biological
Safety Principles
Definitions
 Key Principles
 Data on Laboratory Acquired Infections
 Risk Assessment
 Biosafety Containment Levels
 Primary Containment
 Emergency Preparedness

Introduction to Biosafety
Principles


Complex relationship between organisms and
hosts. We are surrounded by countless
microorganisms. Our bodies depend upon them
for natural processes such as digestion. But most
of time, we do not get sick because of natural
defenses.
Infectious (pathogenic) organisms must:
– Attach and survive hosts defenses
– Multiply
– Create signs and symptoms of disease in host
Definitions
Safety
 Risk
 Biosafety
 Biosecurity

Safety
Freedom from harm
 Control of accidental losses involving

– People
– Property
– Loss to process
Risk

The chance that something may or may not
happen. Often defined as:
– Frequency (how often)
– Severity (how bad)
Biosafety

“Development and implementation of
administrative policies, work practices,
facility design, and safety equipment to
prevent transmission of biological agents
to workers, other persons or the
environment”
MMWR December 6, 2002
Biosecurity

“Protection of high-consequence microbial
agents and toxins, or critical relevant
information, against theft, or diversion by
those who intend to pursue intentional
misuse”
MMWR December 6, 2002
Koch’s Postulates

1890 Robert Koch Established List of Criteria to
Judge Whether or Not a Given Microbe Was
Responsible for a Given Disease
– The organism must be present in every case of the
disease
– The organism must be isolated from the diseased host
and grown in pure culture
– The specific disease must be reproduced when the
pure culture is inoculated into a healthy susceptible
host
– The organism must be recovered from the
experimentally infected host
Biohazardous Materials


Include All Infectious Organisms (Bacteria,
Chlamydiae, Fungi, Parasites, Prions,
Rickettsias, Viruses) which can cause disease in
humans or cause significant environmental or
agricultural impact.
Materials that may harbor infectious organisms
such as human or primate tissues, fluids, cells,
cell cultures.
Key Principles of Biosafety

Laboratory Practices and Techniques
– Hand Washing Important
– Manipulation of Material to Minimize Aerosols
– Consistent Use of Personal Protective Equipment

Safety Equipment (Primary Barriers)
– Biological Safety Cabinets (BSC’s)

Facility Design and Construction (Secondary
Barriers/Room Design)
– Room Pressure Negative to Corridor
– Controlled Access to Non-Research Personnel

Medical Surveillance
Typical Routes of Entry for Viral
or Bacterial Pathogens
Inhalation
 Ingestion
 Injection

– Needle sticks
– Accidental cuts with sharp objects

Skin or Eye Exposure
Laboratory Acquired Infections
Risk of Laboratory Associated Infections
(LAI’s) is Real
 Historical Examples Include:Brucellosis,
Q Fever, Hepatitis, Typhoid Fever,
Tuberculosis, Hepatitis
 Of the 3921 LAI Only 703 (18%) Caused
by Identifiable Accidents including needle
sticks, broken glass, spills or sprays (R.M.
Pike 1976)

40 Years of Data on Overt LAI’s
Pike, R.M. 1978 Various Classes of Agents
No. of Case
No. of Deaths
No. of Agents
Involved
No. of
Published
Cases
Bacteria
1704
71
37
744
Viruses
1179
55
85
915
Rickettsiae
598
25
8
381
Fungi
354
5
9
313
Chlamydiae
128
10
3
71
Parasites
116
2
17
74
Totals
4079
168
159
2498
Agent
Ten Most Frequently Reported
LAI’s
Pike, R.M. 1978 Past and Present Hazards of Agents
Infection
No. of Cases
No. of Deaths
Brucellosis
426
5
Q Fever
280
1
Hepatitis
268
3
Typhoid Fever
258
20
Tularemia
225
2
Tuberculosis
194
4
Dermatomycosis
162
0
Venezuelan Equine
Encephalitis
146
1
Psittacosis
116
10
Coccidiodomycosis
93
2
Totals
2168
48
University’s Responsibilities



To provide a workplace free of recognized
hazards. UA Policy and OSHA Requirement
To provide training to employees in order to
recognize hazards and to protect employees
against those hazards.
Methods of controlling risk may include:
– Building Design Including Containment Features
(Primary/Secondary)
– Policies/Procedures (SOP’s)
– Personnel Protective Equipment
– Medical Surveillance Programs
Basic Risk Assessment
Framework
Hazard Identification
 Estimate Probability of Occurrence
 Decide on Acceptable and Non-Acceptable
Practices
 Implement Practices
 Monitor

Example of Risk Assessment
for Cell Culture
BELGIAN BIOSAFETY SERVER
http://www.biosafety.be/CU/animalcellcultur
es/mainpage.html
 Bioline International
http://www.bioline.org.br/request?by95008

Risk Assessment Flow Chart
Employee Responsibilities




If you don’t know, ask.
If you have not been trained to do it, don’t!
Follow established biosafety practices and
procedures. Always ask Principal Investigator.
Immediately inform Principle Investigator or
Laboratory Manager if any accidents, spills,
procedural issues/concerns or any questions arise
about your safety or the safety of others.
Biosafety Levels Defined
BSL-1
 BSL-2
 BSL-3
 BSL-4

Biosafety Level One (BSL-1)

BSL-1 Work with Well Characterized Agents Not
Known to Cause Disease in Healthy Adults.
Standard Microbiological
– Open bench tops acceptable with good standard
microbiological practices
– Laboratory not necessarily separated
– Special containment equipment or facility design not
required

Examples include E. coli K-12, Bacillus subtilis
Also Called “Bench Work”.
Biosafety Level Two (BSL-2)

BSL-2 Work with Moderate Potential to Affect
Personnel and Environment. (Herpes, Influenza
viruses, Legionella sp.)
– Personnel are specifically trained to handle pathogenic
agents
– Lab access limited when work is conducted
– Extreme precautions taken when handling
contaminated sharp items (needles, scalpels)
– Appropriate immunizations are administered when
available and baseline serum samples encouraged
– Certain procedures require biological safety cabinets
Biosafety Level Three (BSL-3)

BSL-3 Work May Cause Serious or
Potentially Lethal Disease as a Result of
Exposure to Inhalation Route. (M.
Tuberculosis, Bacillus anthraces)
–
–
–
–
Very specific training
Biosafety Cabinets used
Appropriate PPE and other clothing
Specific engineering and design features
Additional (BSL-3)
Requirements
Immunization and medical surveillance
protocols required
 No open bench work
 Ducted exhaust air ventilation creates
directional airflow from “clean” toward
“contaminated” areas prior to discharge to
outside
 High Efficiency Particulate Air (HEPA)
filters may be required for room exhaust

Biosafety Level Four (BSL-4)

BSL-4
Work with dangerous and exotic
agents which pose a high risk of aerosoltransmitted laboratory infectious and life
threatening disease. Ebola, Marburg,
– Special facility design features required
– All activities confined to Class III biosafety cabinets
(glove boxes), or Class II BSC’s used by workers
using one piece positive pressure personnel suits
ventilated by a life support system
Identifying Biohazard Risks



What am I Working
With? How Can it Cause
Disease and How do I
Protect Myself?
Routes of Entry Include
Inhalation, Ingestion,
Inoculation, Skin and
Eyes
Typical Risks of
Exposure Include
Contaminated Needles,
Mouth-Pipetting,
Splashing, Animal Bites
How to Protect Yourself
Knowledge and Understanding of the
Biohazards You Are Working With
 How Can it Get Onto/Into My Body
 How to Protect Myself (Hierarchy of
Control)

– Containment Equipment
– Techniques
– Personal Protective Equipment
Identifying Biohazard Risk is
Key



Accident/Incident
Preceded Events
Represented Only 18%
of LAI’s
Aerosols, Droplets and
Fomites are Likely
Sources
Lab Techniques With
High Potential for
Exposure Include:
– Centrifuges/Blenders,
Opening Tubes/Bottles,
Syringes/Needles,
Inoculating Loops,
Heating Over Flames
Mammalian Tissue Culture
Work

Risks
– Tissue culture may contain virus or bacteria capable
of spreading to human host
– Integrity of culture may be altered because of
contamination from outside source

How to Reduce Risks to Human and Cell
Culture?
– Manipulation of tissue cultures only under Class II
Biological Safety Cabinets
– Use care when doing any procedure using instruments
that may break skin
– Use proper PPE like latex gloves, eye protection
Class II Biological Safety
Cabinets Explained

Main Function
– Protects Worker
– Protects Work (Tissue Cultures From Microbial
Contaminants, i.e.. Integrity of Cultures)

Features
– High Efficiency Particulate Air (HEPA) Filter
Minimizes Escape of Contaminants Within Cabinet
Into Lab
– HEPA Filtered Air Supply Bathes Work Surface,
Protecting Work

Certified Annually by Facilities Management
Basis of Primary Containment
Isolate the Laboratory Worker from Biological Agent With
Ultra Filtered Directional Air Currents
Class II Type B1 Biological
Safety Cabinet Air Flow/HEPA
Filter Placement
Proper Use of Biosafety
Cabinets

Do’s
– Become familiar about the equipment by reading
users manual and asking PI.
– Keep laboratory doors closed and minimize
movement in front of cabinet to avoid disrupting
airflow. Avoid rapid arm movement in and out of
BSC.
– Decontaminate work surfaces with disinfectant before
and after working in a cabinet according to laboratory
standard operating procedures (SOP’s).
Proper Use of Biosafety
Cabinets

Don’ts
– Do not use cabinets as a permanent storage area for
supplies (disrupts airflow)
– Do not work inside cabinet with UV lamp on, if so
equipped. (skin/eye burns)
– Do not rapidly insert or withdraw arms. (disrupts
airflow)
– Place required equipment or supplies for procedure
inside before beginning work. (minimizes hand/arm
withdrawals which can disrupt airflow)
Eagleston Institute Biosafety
Cabinet Clips
What Does Your Lab Look
Like? Advantages of Good
Housekeeping




Reduces Risk of Slip,
Trip and Falls
Protects Integrity of
Biological Experiments
by Providing Adequate
Space and Reduce
Contamination Potential
Easier to Decontaminate
Surfaces
Saves Time by Being
Able to Find Stuff
Emergency Preparedness

What Should I Do When Things Go Wrong?
– Learn the types of emergencies that could happen



Spills of liquids
Equipment malfunctions
Exposure to potential pathogens through inhalation,
ingestion, skin including eye exposure, needle or other sharps
– Learn how to respond to minimize exposure time and
concentration
– Contact your supervisor to protect your health and
legal rights
Biohazard Spills



Each Lab Required to Have Spill
Decontamination Plan
PI Required to Have Cleanup/Decon Procedure
for Specific Biohazards Found in Lab
If Spill Occurs/General Guidelines
–
–
–
–
Remove affected clothing/gloves
Wash contaminated body areas with soap/H2O
Secure area until cleanup completed
Call UA Risk Management 621-1790 for technical
assistance
Summary


Risks of Working with Biological Materials in Research
are Real
The Risks Can be Managed Through:
– Properly Identifying and Assessing Biological Risks
– Facility Design, Construction and Maintenance (Secondary Barriers)
– Correct Use of Safety Equipment (Primary Barriers) Including
Biological Safety Cabinets
– Good Laboratory Practice and Technique

Additional Resources are Available Through Institutional
Biosafety Committee and Professional Staff
Regulatory Framework
PLS 4/595D /Regulations and
Laboratory Management
Spring Semester, 2007
Mark J. Grushka, M.S., CSP
Manager, Biosafety and Biosecurity
University of Arizona
Part 2 Introduction to
Introduction to Regulatory
Framework
What are the major regulations covering
biosafety?
 How is the University of Arizona
organized for biosafety compliance?
 What are the future implications for
regulatory control of biosafety?

Introduction

The regulatory framework covering
biosafety can be characterized as a
combination of statutes, regulations, rules
and guidelines from various federal and
state agencies, private and public
organizations and other interested parties
such as manufacturers of containment
equipment
Federal Laws
Occupational Health and Safety Act (OSHAct)

Bloodborne Pathogens (29 CFR 1910.1030)

Occupational Exposure to Hazardous Chemicals in
Laboratories (29 CFR 1910.1450)

Personal Protective Equipment
(29 CFR 1910.132-139)

Needlestick Standard
National Institutes of Health
NIH Office of Biotechnology Activities
 NIH Guidelines for Research Involving
Recombinant DNA Molecules
 IBC Resources

USDA

APHIS
USDOT
HazMat Safety
 Hazardous Materials Regulations (49
CFR 100-185)

US EPA
Hazardous Waste
 Microbiology
 IAQ

Select Agents
All individuals who have access to Select
Agents must undergo a Security Risk
Assessment
 Acquisition, use, transfer and disposal of
Select Agents is monitored by
CDC/APHIS through issuance of
registration

How is the UA Organized to
Comply?
Compliance based at Vice President for
Research Office
 Manager of Biosafety and Biosecurity
 Chairman of the Institutional Biosafety
Committee
 Program Coordinator
 Graduate Student

UA and Regulatory Reality
Check

Institution Governed by Many Internal
Policies and External Laws/ Regulations
– Provides a “Road Map” for Establishing and
Monitoring Effectiveness of Biosafety
Program
– Keys to Success
Accountability (Who is in Charge)
 Clear Goals and Objectives
 Periodic Monitoring

Regulations and Guidelines for
Biosafety at UA

Occupational Safety and Health Act (OSHA)
– Blood borne Pathogen Standard (Required Training
for All Employees Who Work With Human
Tissues, Blood or Other Bodily Fluids Must Take
Course From UA Risk Management)




University of Arizona Biosafety Handbook
Biosafety in Microbiological and Biomedical
Laboratories (CDC/NIH) 5th Edition
NIH Guidelines for Research Involving
Recombinant DNA Molecules
Laboratory Specific Procedures (SOP’s)
Institutional Biosafety
Committee Requires a Written
Plan from PI for These Types of
Research
Recombinant DNA
 Pathogenic Microorganisms
 Mammalian Cell Lines
 Gene Therapy
 Transgenic Plants

Institutional Biosafety
Committee Basics
Reports to Vice President for Research
 Insures a safe working environment by
minimizing exposure of personnel to
harmful biological agents
 Peer Review of research conducted at or
sponsored by the U of A for compliance
with adopted policies, regulations and
guidelines

Where to Get More Information
IBC Website http://www.ibc.arizona.edu/
 Risk Management http://risk.arizona.edu/
 CDC Website http://cdc.gov
 OSHA Websitehttp://labor/osha.gov

Mark J. Grushka, Manager, Biosafety and
Biosecurity 621-5279 and Margaret Stalker,
Program Coordinator 621-3441
Memorandum of Understanding
and Agreement Form
Is the main risk assessment document
submitted by Principal Investigators
 Submitted in on-line form
 Reviewed at least twice

– Prereview
– Committee Review

Approval from IBC allows PI to conduct
research at specific BSL level
Memorandum of Understanding
and Agreement Form (MUA)
On-line Submission
 Provides Key Information for Institutional
Biosafety Committee Review
 Provides Documentation that PI Will
Adhere to Specific Levels of Control for
Biological Risk Management as Part of the
Conduct of Research Project

MUA New User Profile: Administrative
and Emergency Information
SECTION 1: Project Information
Auditing Function
All BSL-3 laboratories audited annually by
Manager of Biosafety
 All Select Agent laboratories audited
annually by Manager of Biosafety
 All BSL-1 applications require an on-site
inspection prior to consideration by IBC

Other Resources

Training
– Bloodborne Pathogen and Shipping of
Hazardous Materials by Air done by Risk
Management and Safety
On-line UA Biosafety Handbook
 Program Manager,Manager of Biosafety
and IBC Chair available to respond to
technical or regulatory questions

Guidance Documents




World Health Organization Biosafety
Manual http://www.who.int/csr/delibepidemics/
WHO_CDS_CSR_LYO_2004_11/en/
2nd Edition Primary Containment for
Biohazards:Selection, Installation and
Use of Biological Safety Cabinets
http://www.cdc.gov/od/ohs/biosfty/bsc/bsc.htm
Biosafety in Microbiological and
Biomedical Laboratories (BMBL) 4th
Edition
http://www.cdc.gov/od/ohs/biosfty/bmbl4/bmbl4t
oc.htm
American Biological Safety Association
(ABSA)
http://www.absa.org/restool.html
Regulatory Summary

Compliance Environment Characteristics
– Combination of laws, regulations and
guidance documents
– Objective is to reduce chance of impact of
biological research on occupational health,
public health and environmental impacts

Institution has significant autonomy in
terms of defining how biosafety is
organized to meet compliance objectives
Scenario 1:
The Dedicated Post-Doc
 Situation:
– A new Post-Doc has just arrived from a prestigious University. He is
working on improving the production of Interferon from a human
cell line. Unfortunately the cell line also produces Human T-cell
leukemia virus. He brings this cell line in from the University where
he was working before and decides to grow it up in incubators in
several labs without telling anyone else working in those labs that it is
also co-contaminated with HTLV III.
 What
 How
do you do?
can this have been avoided?
Slide 21
Scenario 2:
The Helpful Lab Worker
 Situation:
– A lab assistant in a private lab is working with a mouse cell line that
has been genetically modified to produce the entire genome of HIV
without the LTR sequences (so it is non-infectious). She has been
contacted by a very prestigious colleague from another private
institution in Europe and was invited to visit their lab. She decides to
take along a vial of her cell line packaged in liquid nitrogen. To make
sure there is no delay she is hand carrying the vial on the plane in her
carry-on bag.
 Is
this wrong?
 Why?
 How
can this have been avoided?
Slide 22
Scenario 3:
The Reluctant Director

Situation:
– Your institution is just implementing a new Biosafety Program. Up
until now no one has ever questioned the Laboratory Animal Sciences
(LAS) Department about their work. The Director of LAS is
reluctant to have his department participate in this newfangled
program that is only going to hamper his group and make life more
difficult for him and his people.

What can you do to ensure that LAS
participates in the program?
Slide 23
Scenario 4:
The Busy Scientist
 Situation:
– A Nobel winning scientist is working in your company. He is a very
important person and is much too busy to attend any Biosafety
training sessions or let his assistants attend. Besides, what could you
possibly show him?
 What
do you do?
 How
can you convince him of the need to
attend training?
 How
can this have been avoided?
Slide 24