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I'll
IN THE LABORATORY
Prudent Practices for the Handling
and Disposal of Infectious Materials
Committee on Hazardous Biological Substances in the Laboratory
Board on Chemical Sciences and Technology
Commission on Physical Sciences, Mathematics, and Resources
National Research Council
NATIONAL ACADEMY PRESS
Washington, D.C. 1989
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NATIONAL ACADEMY PRESS ~
2101 Constitution Avenue, NVV
.
Washington, DC 20418
NOTICE: The project that is the subject of this report was approved by the Goveming Board of the National Academy of Sciences, the
National Academy of Engineenng, and the Institute of Medicine. The members of the committee responsible for the report were chosen
for their special competences and with regard for appropriate balance.
This report has been reviewed by a group other than the authors according to procedures approved by a Report Review Committee
consisting of members of the National Academy of Sciences, the National Academy of Engineenng, and the Institute of Medicine.
The National Academy of Sciences is a private, nonprofit, self-perpetuating society of distinguished scholars engaged in scientific
and engineering research, dedicated to the furtherance of science and technology and to their use for the general welfare. Upon the
authority of the charter granted to it by the Congress in 1863, the Academy has a mandate that requires it to advise the federal government
on scientific and technical matters. Dr. Frank Press is the president of the National Academy of Sciences.
The National Academy of Engineering was established in 1964, under the charter of the National Academy of Sciences, as a parallel
organization of outstanding engineers. It is automonous in its administration and in the selection of its members, sharing with the National
Academy of Sciences the responsibility for advising the federal govemment. The National Academy of Engineering also sponsors
engineering programs aimed at meeting national needs, encourages education and research, and recognizes the superior achievements of
engineers. Dr. Robert M. White is president of the National Academy of Engineering.
The Institute of Medicine was established in 1970 by the National Academy of Sciences to secure the services of enunent members of
appropriate professions in the examination of policy matters pertaining to the health of the public. The Institute acts under the
responsibility given to the National Academy of Sciences by its congressional charter to be an adviser to the federal government and, upon
its own initiative, to identify issues of medical care, research, and education. Dr. Samuel O. Thier is president of the Institute of Medicine.
The National Research Council was organized by the National Academy of Sciences in 1916 to associate the broad community of
science and technology with the Academy's purpose of furthering knowledge and advising the federal govemment. Functioning in
accordance with general policies detemiined by the Academy, the Council has become the principal operating agency of both the National
Academy of Sciences and the National Academy of Engineering in providing services to the govemment, the public, and the scientific and
engineering communities. The Council is administered jointly by both Academies and the Institute of Medicine. Dr. Frank Press and Dr.
Robert M. White are chairman and vice chairman, respectively, of the National Research Council.
Support for this project was provided by the U.S. Department of Agriculture and National Institutes of Health under contract no. 59-
32U4-5-33; the U.S. Department of the Army under contract no. DAMD17-86~ 6022; the U.S. Department of Energy under grant no. DE-
FG05-85ER13457; the U.S. Department of Labor under purchase order not B9P56292; and the National Science Foundation under "rant no.
DMB-8611553. The project has also been supported by funds from the National Research Council Fund, a pool of private, discretionary,
nonfederal funds that is used to support a program of Academy-initiated studies of national issues in which science and technology figure
significantly. The NRC Fund consists of contributions from a consortium of private foundations including the Camegie Corporation of New
York, the Charles E. Culpeper Foundation, the William and Flora Hewlett Foundation, the John D. and Catherine T. MacArthur Foundation,
the Andrew W. Mellon Foundation, the Rockefeller Foundation, and the Alfred P. Sloan Foundation, as well as the Academy Industry
Program, which seeks annual contributions from companies that are concerned with the health of U.S. science and technology and with
public policy issues with technological content.
Library of Congress Cataloging-in-Publication Data
National Research Council (U.S.). Committee on Hazardous Biologic
Substances in the Laboratory.
Biosafety in the laboratory: prudent practices for the handling and
disposal of infectious materials / Committee on Hazardous Biological
Substances in the Laboratory, Board on Chemical Sciences and
Technology, C~issian on Physical Sciences, Mathematics, and
Resources, National Research Council.
p. cm.
Includes bibliographical references.
ISBN 0-309~39754
1. Medical laboratories-Safety measures~ngresses. 2. Medical
laboratories Waste disposal~afety measures-Congresses.
I. Title.
[DNLM: 1. Containment of Biohazards-standards United States.
2. Laboratories standards United States. 3. LaboratoryInfection-
prevention dc control United States. QY 23 N2775b]
R860.N37 1989
616.9'0072~1c20
DNI~I/DLC for library of Congress
Copyright ~ 1989 by the National Academy of Sciences
Printed in the United States of America
89-13004
CIP
First Printing, November 1989
Second Pnnting, July 1990
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Committee on Hazardous Biological
Substances in the Laboratory
Committee (Authors)
EDWARD A. ADELBERG (Chairman), Yale University
ROBERT AUSTRIAN, University of Pennsylvania
School of Medicine
HOWARD L. BACHRACH, Agricultural Research
Service, U.S. Department of Agriculture
W. EMMETT BARKLEY, National Institutes of Health
J. PAUL BURNET, Lilly Research Laboratories
DIANE O. FLEMING, Sterling Research Group
ROY L. FUCHS, Monsanto Company
HAROLD S. GINSBERG, Columbia University
ROSE GOLDMAN, Cambridge Hospital, Harvard
University School of Medicine
JAMES M. HUGHES, Centers for Disease Control
WILLIAM G. MIKELL, E.I. du Pont de Nemours &
Company, Inc.
JOHN H. RICHARDSON, Emory University
JEROME P. SCHMIDT, USAF School of Aerospace
Medicine
JAMES W. SMITH, Indiana University School of
Medicine
THOMAS E. WALTON, Agricultural Research Service,
U.S. Department of Agriculture
·~e
NRC Staff
WILLIAM SPINDEL, Study Director (July 1985 to
January 1987)
ROBERT M. SIMON, Study Director (January 1987 to
July 1989)
BENNETT L. ELISBERG, Consultant
MONALISA BRUCE, Administrative Secretary
SANDRA NOLTE, Administrative Secretary
Contributors
MICHELLE EVANS, National Institutes of Health
GREGG J. HUNT, Agricultural Research Service, U.S.
Department of Agriculture
JOHN KEENE, Abbott Laboratories
GEORGE P. KUBICA, Centers for Disease Control
ROBERT W. McKINNEY, National Institutes of Health
JONATHAN RICHMOND, National Institutes of Health
HARVEY W. ROGERS, National Institutes of Health
CLARENCE STYRON, Monsanto Company
JERRY TULIS, University of Norm Carolina
DONALD VESLEY, University of Minnesota
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Board on Chemical Sciences and Technology
EDWARD A. MASON (Co-Chairman), Amoco
Corporation
GEORGE M. WHITESIDES (Co-Chairman), Harvard
University
ALEXIS T. BELL, University of California, Berkeley
JOHN I. BRAUMAN, Stanford University
PETER B. DERVAN, California Institute of Technology
GARY FELSENFELD, National Institutes of Heals
JEANETTE G. GRASSELLI, Ohio State University
MICHAEL L. GROSS, University of Nebraska
RALPH HIRSCHMANN, University of Pennsylvania
ROBERT L. LETSINGER, Northwestem University
JAMES F. MATHIS, Exxon Chemical Company (retired)
JOHN A. QUINN, University of Pennsylvania
STUART A. RICE, University of Chicago
iv
FREDERIC M. RICHARDS, Yale University
GERALDINE L. RICHMOND, University of Oregon
ROGER A. SCHMrl2, University of None Dame
L.E. SCRIVEN, University of Minnesota
DAVID P. SHEETS, Dow Chemical USA
LEO J. THOMAS, JR., Sterling Drug Company
NICHOLAS J. TURRO, Columbia University
MARK S. WRIGHTON, Massachusetts Institute of
Technology
ROBERT M. SIMON, StaffDirector
WILLIAM SPINDEL, SpecialSta~Advisor
PEGGY J. POSEY, StaffAssociate
LYNN E. DUFF, Financial Assistant
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Commission on Physical Sciences
Mathematics, and Resources
NORMAN HACKERMAN (Chairman), Robert A. Welch
Foundation
GEORGE F. CARRIER, Harvard University
HERBERT D. DOAN, The Dow Chemical Company
(retired)
PETER S. EAGLESON, Massachusetts Institute of
Technology
DEAN E. EASTMAN, IBM T.J. Watson Research Center
MARYE ANNE FOX, University of Texas, Austin
GERHART FRIEDLANDER, Brookhaven National
Laboratory
LAWRENCE W. FUNKHOUSER, Chevron Corporation
(retired)
PHILLIP A. GRIFFITHS, Duke University
CHRISTOPHER F. McBE, University of California,
Berkeley
JACK E. OLIVER, Cornell University
JEREMIAH P. OSTRIKER, Princeton University
Observatory
FRANK L. PARBR, Vanderbilt University
DENIS J. PRAGER, MacArthur Foundation
DAVID M. RAUP, University of Chicago
RICHARD J. REED, University of Washington
ROY F. SCHWITTERS, Superconducting Super Collider
Laboratory
ROBERT E. SIEVERS, University of Colorado
LEON T. SILVER, California Institute of Technology
LARRY L. SMARR, University of Illinois
EDWARD C. STONE, JR., California Institute of
Technology
KARL K. TUREKIAN, Yale University
MYRON F. UMAN, Acting Executive Director
V
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Preface
In 1981 and 1983, the National Research Council published two reports on
chemical safety in the laboratory: Prudent Practices for Handling Hazardous
Chemicals in Oratories and Prudent Practices for Disposal of Chemicals from
Laboratories [93,941. In November 1983, a planning committee of the National
Research Council was convened under the chairmanship of Thomas Weller to
consider the need for a document that would deal in a similar way with biological
safety in the laboratory. The committee concluded that such a document would be
timely and recommended the formation of a working committee to produce it.
The Committee on Hazardous Biological Substances in the Laboratory was
organized in the fall of 1985. It was presented with a broad charge, namely, to
prepare a report dealing with the following aspects of hazardous hiolo~ical mam-
rials:
als.
--cat ~
to
1. Definition of laboratory safety problems with hazardous biological maten
2. Guidelines for physical facilities, equipment, and word` practices.
3. Procedures for identifying hazards and establishing conditions for any
operation involving hazardous biological matenals.
4. Guidelines for waste disposal, including incinerating, venting, and dis-
charging to sewer systems.
5. Guidelines for all aspects of an effective safety program including medical
surveillance, compliance with regulations, and recor~keeping.
6. A plan for obtaining consensus on and implementation of the guidelines.
The committee first met in January 1986 and decided to restrict the scope of
the report to the safe handling and disposal of agents hazardous to humans; strict
animal pathogens and strict plant pathogens were considered to be of interest to
different, specialized audiences, and better dealt with in other publications. It was
also decided to deal only briefly with such hazardous biological products as toxins
and immunoactive substances.
During the period in which this report was being planned and written, a
number of excellent books appeared dealing with various aspects of biosafety in
the laboratory (see, for example, references 4, 83, and 149~. Although the present
report overlaps many sections of these books, the committee felt that the need still
vii
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·~e
V111
PREFACE
existed for a consensus, peer-reviewed document, produced under the imprimatur
of the NRC, that could serve as a general set of guidelines for the safe handling
and disposal of infectious materials in the laboratory. This book represents the
committee's efforts to produce such a document, with the able support of the
National Research Council's staff: in particular, we wish to thank William Spin-
del, Robert M. Simon, and Bennett L. Elisberg for their expert assistance. We also
wish to acknowledge the contributions of the 30 or more reviewers, representing
every type of microbiological laboratory, whose thoughts and constructive
comments formed the basis of many changes in the final draft of this book.
EDWARD A. ADELBERG, Chairman
Committee on Hazardous Biological
Substances in the Laboratory
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Contents
INTRODUCTION, OVERVIEW, AND RECOMMENDATIONS /1
A. INTRODUCTION.....
B. OVERVIE W ................
C. RECOMMENDATIONS.......
DESCRIPTIVE EPIDEMIOLOGY OF OCCUPATIONAL INFECTIONS OF
LABORATORY WORKERS / 8
A. INTRODUCTION...........
B. THE EPIDEMIOLOGIC TRIAD
1. The Host/8
2. The Infectious Agent / 8
3. The Environment / 9
C. LABORATORY-ASSOCIATED INFECTIONS
1. Infectious Agents Presenting the Highest Risk / 9
2. Infectious Agents Presenting the Lowest Risk / 11
3. Other Infectious Agents / 12
. .
3
SAFE HANDLING OF INFECTIOUS AGENTS / 13
8
8
· · 9
A. GUIDELINES FOR HANDLING PATHOGENIC MICROORGANISMS. . 13
B. ORGANISMS POSING SPECIAL RISKS 1 3
C. HAZARDS FROM VERTEBRATE ANIMALS AND INSECTS IN THE
LABORATORY .......................................
D. PRIMARY AND CONTINUOUS CELL CULTURES ........
E. HANDLING OF NECROPSY AND SURGICAL SPECIMENS .
1. Introduction / 16
2. Necropsy/16
a. Routine necropsies, 16; b. Necropsies on bodies known to be infected, 16
3. Surgical Pathology / 18
ix
14
... 15
16
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x
CONTENTS
F. GOOD LABORATORY PRACTICES
1. In~oduction/18
2. Routes of Exposure / 18
a. contact route, 18; b. Cal route, 19; c. Ocular route, 19; d. laaculation route, 19;
e. Respiratory route, 19
3. Prevention of Exposure / 19
4. The Seven Basic Rules of Biosafety / 19
5. Summary / 20
G. TRANSPORTATION AND SHIPMENT OF SPECIMENS
1. Introduction / 20
2. Packing, Shipping, and Handling of Biological Specimens / 20
H. LABELING OF SPECIMENS WITHIN THE LABORATORY
I. PREVENTION OF AEROSOL AND DROPLET GENERATION
1. Introduction / 22
2. Control of Aerosols and Droplets / 23
J. CONTAINMENT EQUIPMENT
1. In~oduction/25
2. Biological Safety Cabinets / 25
3. Pipethng Devices / 26
4. Sonicators, Homogenizers, and Mixers / 26
5. Clothing, Masks, and Face Shields / 28
K. BIOSAFETY IN LARGE-SCALE PRODUCTION
1. In~oduction/28
2. Organization and Responsibilities / 30
3. Containment/30
4. Inactivation / 31
5. Disposal / 32
6. Exposure / 32
7. Conclusion / 33
20
22
L. BIOSAFETY IN PHYSICIANS' OFfilCE LABORATORIES AND
OTHER SMALL-VOLUME CLINICAL LARORATOR]F.R
4
SAFE DISPOSAL OF INFECTIOUS LABORATORY WASTE / 34
33
A. INTRODUCTION en · _~
B. INFECTIOUS POTENTIAL OF LABORATORY WASTE 35
1. Risks to the General Public's Health / 35
2. Occupational Risks / 36
C. CHARACTERISTICS OF INFECTIOUS LABORATORY WASTE 36
D. RESPONSIBILITY FOR THE SAFE HANDLING AND
DISPOSAL OF INFECTIOUS WASTE - - - 16
1. Generators of Infectious Waste / 37
2. Haulers and Waste Treatment Facilities / 37
WASTE HANDLING AND TREATMENT METHODS .
1. Basic Principles / 37
2. Containment / 38
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CONTENTS
Xl
3. Personal Protection / 39
4. Chemical Decontamination / 39
5. Steam Autoclaving / 39
6. Incineration/41
7. Validation of Decontamination Methods / 42
INFECT lOUS WASTE PIQUING SPECIAL CONSIDERATION
1. Mixed Waste / 42
2. Human Cadavers and Other Anatomical Waste / 44
3. Animal Bedding Materials / 44
4. ''Shalps''/44
SAFETY MANAGEMENT / 46
A. ADMINISTRATIVE ORGANIZATION AND RESPONSIBILITIES
1. Introduction/46
2. The Laboratory Safety Program / 46
a. Goals of a laboratory safety program, 46; b. Responsibility for laboratory
safety, 46; c. Safety plans, 47; d. Safety meetings and safety committees, 47;
e. Safety communications, 48; f. Monitoring safety, 48
B . FA FIT ITIFS
1. Introduction / 48
2. Laboratory Design /49
a.Ventilation, 49; b. Electrical, 49; c. Water, 50; d. Sewage, 50; e. Vacuum, 50;
I. Waste handling, 50; g. Safety equipment, 50; h. Traffic flow pattern, 50;
i. Laundry, 50; i. Storage areas, 51
3. Constructing, Remodeling, and Decommissioning a Laboratory / 51
4. Maintenance/51
5. Housekeeping / 51
C. OPERATIONS.................................................
1. Introduction / 51
2. Safety Orientation and Continuing Education for Employees / 52
3. Evaluation of Laboratory-Associated Hazards / 53
4. Policy and Procedure Manuals / 53
5. Accident Reports and Investigations / 54
6. Recordkeeping / 54
7. Auditing / 54
8. Registry of Agents / 55
9. Waste Management / 55
10. Signs/55
D. hDEDICAiL PROGRAl4................................
1. General Principles / 55
2. Conditions Increasing Employee Risk of Adverse Health Outcome / 55
a. Deficiencies of host defenses, 56; b. Reproductive hazards, 56; c. Allergies, 57
3. Program Design / 57
4. Preplacement Evaluation (PPE) / 57
a. Medical history, 57; b. Occupational health history, 57;
d. Laboratory and other testing, 58; e. Serum bank 58
c.
· · .
42
46
48
51
· · . . . · . . . . 55
Physical examination, 57;
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·.
X11
CONTENTS
5. Immunizations / 58
6. Periodic Monitoring Examination (PME) / 58
7. Postemployment Evaluation (PEE) / 59
Agent-SpeciD~c Surveillance / 59
Accident Reporting / 59
8.
9.
10. Recordkeeping and Result Notification / 59
11. Resources / 59
E EMERGENCES ,~ en eee~eeeee ~--VJ
1. Preparation and General Procedures / 63
a. Preparation, 63; b. General emergency procedures, 63
2. Evacuation Procedures / 63
a. Emergency alarm system, 63; b. Evacuation routes, 64; c. Shutdown procedures, 64;
d. Start-up procedures, 64; e. Drills, 64; f. Power failure, 64
3. Fires / 64
4. Spills and Releases / 64
a. Infectious agents, 64; b. Handling of spilled solids, 66; c. Biological radioactive
emergencies, 66
5. Other Emergencies / 67
F. REGULATION AND ACCREDITATION v,
G. TEACHING BIOSAFETY IN ACADEMIC SETTINGS .
1. In~oduction/67
2. Safety in Laboratory Courses / 68
3. Orientation and Training of Students / 68
4. Design of Safe Laboratory Exercises and Experiments / 68
5. Monitoring and Recordkeeping / 69
REFERENCES / 70
APPENDIXES
A. Biosafety in Microbiological and Biomedical Laboratories
B. 1988 Agent Summary Statement for HIVs, Including HTLV-III,
LAV, HIV-l, and HIV-2
83
......... 141
C. Recommendations for Prevention of HIV Transmission in
Heallh-Care Settin~.s
155
D. Summary of Zoonotic Pathogens Causing Disease in Man 175
E. Regulations Governing the Packaging, Labeling, and Transport of
Infectious Agents 187
F. Teaching Aids and Training Courses 205
G. Regulation and Accreditation. - -
H. List of Abbreviations .......
INDEX / 213
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.~ ~'7~1%
IN THE LABORS
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