Third edition - who.int

1 World Health OrganizationGeneva2004 Laboratory biosafety manualThird editionWHO Library Cataloguing-in-Publication DataWorld Health biosafety manual. 3rd of biohazards - methods - standards - prevention and control 92 4 154650 6(LC/NLM classification: QY 25)WHO/CDS/CSR/ World Health Organization 2004 All rights reserved. Publications of the World Health Organization can be obtained from Marketing andDissemination, World Health Organization, 20 Avenue Appia, 1211 Geneva 27, Switzerland (tel: +41 22791 2476; fax: +41 22 791 4857; email: Requests for permission to reproduce ortranslate WHO publications whether for sale or for noncommercial distribution should be addressedto Publications, at the above address (fax: +41 22 791 4806.))

2 Email: designations employed and the presentation of the material in this publication do not imply theexpression of any opinion whatsoever on the part of the World Health Organization concerning the legalstatus of any country, territory, city or area or of its authorities, or concerning the delimitation of itsfrontiers or boundaries. Dotted lines on maps represent approximate border lines for which there may notyet be full mention of specific companies or of certain manufacturers products does not imply that they areendorsed or recommended by the World Health Organization in preference to others of a similar naturethat are not mentioned. Errors and omissions excepted, the names of proprietary products are distinguishedby initial capital World Health Organization does not warrant that the information contained in this publication iscomplete and correct and shall not be liable for any damages incurred as a result of its by minimum graphicsPrinted in MaltaThis publication was supported by Grant/Cooperative Agreement Number U50/CCU012445-08from the Centers for Disease Control and Prevention (CDC), Atlanta, GA, USA.

3 Its contents aresolely the responsibility of the authors and do not necessarily represent the official views of iii ForewordviiAcknowledgementsviii1. General principles1 Introduction1 PART I. biosafety guidelines52. Microbiological risk assessment7 Specimens for which there is limited information8 Risk assessment and genetically modified microorganisms83. Basic laboratories biosafety Levels 1 and 29 Code of practice9 Laboratory design and facilities12 Laboratory equipment14 Health and medical surveillance16 Training16 Waste handling17 Chemical, fire, electrical, radiation and equipment safety194. The containment laboratory biosafety Level 320 Code of practice20 Laboratory design and facilities21 Laboratory equipment22 Health and medical surveillance225.

4 The maximum containment laboratory biosafety Level 425 Code of practice25 Laboratory design and facilities256. Laboratory animal facilities28 Animal facility biosafety Level 129 Animal facility biosafety Level 229 Animal facility biosafety Level 330 Animal facility biosafety Level 431 Invertebrates327. Guidelines for laboratory/facility commissioning338. Guidelines for laboratory/facility certification36 PART II. Laboratory biosecurity459. Laboratory biosecurity concepts47 PART III. Laboratory equipment4910. Biological safety cabinets51 Class I biological safety cabinet51 Class II biological safety cabinets53 Class III biological safety cabinet56 Biological safety cabinet air connections56 Selection of a biological safety cabinet57 Using biological safety cabinets in the laboratory5711.

5 Safety equipment61 Negative-pressure flexible-film isolators61 Pipetting aids63 Homogenizers, shakers, blenders and sonicators63 Disposable transfer loops64 Microincinerators64 Personal protective equipment and clothing64 PART IV. Good microbiological techniques6712. Laboratory techniques69 Safe handling of specimens in the laboratory69 Use of pipettes and pipetting aids70 Avoiding the dispersal of infectious materials70 Use of biological safety cabinets70 Avoiding ingestion of infectious materials and contact with skin and eyes71 Avoiding injection of infectious materials71 Separation of serum72 Use of centrifuges72 Use of homogenizers, shakers, blenders and sonicators73 Use of tissue grinders73 Care and use of refrigerators and freezers73 Opening of ampoules containing lyophilized infectious materials74 Storage of ampoules containing infectious materials74 Standard precautions with blood and other body fluids, tissues and excreta74 Precautions with materials that may contain prions7613.

6 Contingency plans and emergency procedures78 Contingency plan78 Emergency procedures for microbiological laboratories7914. Disinfection and sterilization82 Definitions82 Cleaning laboratory materials83 iv LABORATORY biosafety MANUALC hemical germicides83 Local environmental decontamination88 Decontamination of biological safety cabinets89 Hand-washing/hand decontamination90 Heat disinfection and sterilization90 Incineration92 Disposal9315. Introduction to the transport of infectious substances94 International transport regulations94 The basic triple packaging system95 Spill clean-up procedure95 PART V. Introduction to biotechnology9916. biosafety and recombinant DNA technology101 biosafety considerations for biological expression systems102 biosafety considerations for expression vectors102 Viral vectors for gene transfer102 Transgenic and knock-out animals102 Transgenic plants103 Risk assessments for genetically modified organisms103 Further considerations104 PART VI.

7 Chemical, fire and electrical safety10517. Hazardous chemicals107 Routes of exposure107 Storage of chemicals107 General rules regarding chemical incompatibilities107 Toxic effects of chemicals107 Explosive chemicals108 Chemical spills108 Compressed and liquefied gases10918. Additional laboratory hazards110 Fire hazards110 Electrical hazards111 Noise111 Ionizing radiation111 PART VII. Safety organization and training11519. The biosafety officer and biosafety committee117 biosafety officer117 biosafety committee118 v CONTENTS20. Safety for support staff119 Engineering and building maintenance services119 Cleaning (domestic) services11921. Training programmes120 PART VIII. Safety checklist12322. Safety checklist125 Laboratory premises125 Storage facilities125 Sanitation and staff facilities126 Heating and ventilation126 Lighting126 Services126 Laboratory biosecurity127 Fire prevention and fire protection127 Flammable liquid storage128 Compressed and liquefied gases128 Electrical hazards128 Personal protection129 Health and safety of staff129 Laboratory equipment130 Infectious materials130 Chemicals and radioactive substances130 PART IX.

8 References, annexes and index133 References135 Annex 1 First aid138 Annex 2 Immunization of staff139 Annex 3 WHO biosafety Collaborating Centres140 Annex 4 Equipment safety141 Equipment that may create a hazard141 Annex 5 Chemicals: hazards and precautions145 Index170 vi LABORATORY biosafety MANUALF oreword vii The World Health Organization (WHO) has long recognized that safety and, inparticular, biological safety are important international issues. WHO published thefirst edition of the Laboratory biosafety manual in 1983. The manual encouragedcountries to accept and implement basic concepts in biological safety and to developnational codes of practice for the safe handling of pathogenic microorganisms inlaboratories within their geographical borders.

9 Since 1983, many countries have usedthe expert guidance provided in the manual to develop such codes of practice. A secondedition of the manual was published in continues to provide international leadership in biosafety through this thirdedition of the manual by addressing biological safety and security issues facing us inthe current millennium. The Third edition stresses throughout the importance ofpersonal responsibility. New chapters have been added on risk assessment, safe use ofrecombinant DNA technology and transport of infectious materials. Recent worldevents have revealed new threats to public health through deliberate misuse and releaseof microbiological agents and toxins. The Third edition therefore also introducesbiosecurity concepts the protection of microbiological assets from theft, loss ordiversion, which could lead to the inappropriate use of these agents to cause publichealth harm.

10 This edition also includes safety information from the 1997 WHOpublication Safety in health-care laboratories (1).The Third edition of the WHO Laboratory biosafety manual is a helpful referenceand guide to nations that accept the challenge to develop and establish national codesof practice for securing microbiological assets, yet ensuring their availability for clinical,research and epidemiological A. Asamoa-BaahAssistant Director-GeneralCommunicable DiseasesWorld Health OrganizationGeneva, SwitzerlandAcknowledgements viii The development of this Third edition of the Laboratory biosafety manual has beenmade possible through the contributions of the following, whose expertise is gratefullyacknowledged:Dr W. Emmett Barkley, Howard Hughes Medical Institute, Chevy Chase, MD, USADr Murray L.