WHO/CDS/CSR/LYO/2003.4 LABORATORY BIOSAFETY MANUAL SECOND EDITION (REVISED) interim guidelines WORLD HEALTH ORGANIZATION Geneva 2003 interim guidelines © World Health Organization 2003 This publication is not a formal publication of the World Health Organization (WHO), and all rights are reserved by the Organization. The publication may, however, be freely reviewed, abstracted, reproduced and translated, in part or in whole, but not for sale or for use in conjunction with commercial purposes. The views expressed in publications by named authors are solely the responsibility of those authors. The designations employed and the presentation of the material in this publication, including tables and maps, do not imply the expression of any opinion whatsoever on the part of the secretariat of the World Health Organization concerning the legal status of any country, territory, city or area or of its authorities, or concerning the delimitation of its frontiers or boundaries. Dotted lines on maps represent approximate border lines for which there may not yet be full agreement. The mention of specific companies or of certain manufacturers' products does not imply that they are endorsed or recommended by WHO in preference to others of a similar nature that are not mentioned. Errors and omissions excepted, the names of proprietary products are distinguished by initial capital letters. WHO/CDS/CSR/LYO/2003.4 Contents Foreword......................................................................................................................................................v Acknowledgements ..................................................................................................................................vii 1. General principles............................................................................................................................... 1 PART I Guidelines 2. Risk assessment................................................................................................................................. 7 Specimens for which there is limited information .............................................................................. 7 Risk assessment and genetically modified microorganisms ............................................................. 8 3. Basic laboratories – Biosafety Levels 1 and 2............................................................................... 10 Code of practice............................................................................................................................... 10 Laboratory design and facilities ....................................................................................................... 11 Laboratory equipment...................................................................................................................... 13 Health and medical surveillance ...................................................................................................... 13 Training ............................................................................................................................................ 14 Waste handling ................................................................................................................................ 14 Chemical, fire, electrical and radiation safety .................................................................................. 16 4. The containment laboratory – Biosafety Level 3........................................................................... 17 Code of practice............................................................................................................................... 17 Laboratory design and facilities ....................................................................................................... 17 Laboratory equipment...................................................................................................................... 18 Health and medical surveillance ...................................................................................................... 18 5. The maximum containment laboratory – Biosafety Level 4 ......................................................... 20 Laboratory design and facilities ....................................................................................................... 20 Laboratory biosafety manual ........................................................................................................... 21 6. Laboratory animal facilitiesinterim ................................................................................................. guidelines ............ 22 Animal facility – Biosafety Level 1 ................................................................................................... 22 Animal facility – Biosafety Level 2 ................................................................................................... 23 Animal facility – Biosafety Level 3 ................................................................................................... 23 Animal facility – Biosafety Level 4 ................................................................................................... 24 Invertebrates .................................................................................................................................... 24 PART II Laboratory equipment 7. Biological safety cabinets................................................................................................................ 29 Class I biological safety cabinet....................................................................................................... 30 Class II biological safety cabinets.................................................................................................... 30 Class III biological safety cabinet..................................................................................................... 32 Biological safety cabinet air connections......................................................................................... 33 Selection of a biological safety cabinet............................................................................................ 33 Using biological safety cabinets in the laboratory............................................................................ 34 8. Equipment-related hazards.............................................................................................................. 37 Equipment that may create a hazard............................................................................................... 37 i Laboratory Biosafety Manual, 2nd revised edition WHO/CDS/CSR/LYO/2003.4 9. Equipment designed to reduce biological hazards....................................................................... 39 Negative-pressure flexible-film isolators.......................................................................................... 40 Pipetting aids ................................................................................................................................... 40 Homogenizers, shakers, blenders and sonicators........................................................................... 41 Disposable transfer loops ................................................................................................................ 41 Microincinerators.............................................................................................................................. 41 Personal protective clothing and equipment.................................................................................... 41 PART III Good microbiological technique 10. Safe laboratory techniques.............................................................................................................. 45 Safe handling of specimens in the laboratory.................................................................................. 45 Use of pipettes and pipetting aids.................................................................................................... 45 Avoiding the dispersal of infectious materials..................................................................................46 Use of biological safety cabinets ..................................................................................................... 46 Avoiding ingestion of infectious materials and contact with skin and eyes ..................................... 46 Avoiding injection of infectious materials......................................................................................... 47 Separation of serum ........................................................................................................................ 47 Use of centrifuges............................................................................................................................ 47 Use of homogenizers, shakers, blenders and sonicators................................................................ 48 Use of tissue grinders ...................................................................................................................... 48 Care and use of refrigerators and freezers......................................................................................48 Opening of ampoules containing lyophilized infectious materials ................................................... 48 Storage of ampoules containing infectious materials .....................................................................