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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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