Biological Safety BSL3 Laboratory Manual Yale Environmental Health & Safety Yale University - Biological135 College Safety BSL3 Street, Laboratory Suite Manual 100, New Haven, CT 06510 Revised September 2019 ii Yale University Environmental Health & Safety Telephone: 203-785-3550 / Fax: 203-785-7588 Revised September 2019 SECTION 1 RESPONSIBILITIES ..................................................................................................................... 1 1.1 DEPARTMENT CHAIR ...................................................................................................................................... 1 1.2 PRINCIPAL INVESTIGATOR ............................................................................................................................. 1 1.3 RESEARCH PERSONNEL .................................................................................................................................. 2 1.4 ENVIRONMENTAL HEALTH AND SAFETY (EHS) ........................................................................................... 2 1.5 BIOLOGICAL SAFETY COMMITTEE ................................................................................................................ 3 SECTION 2 MEDICAL SURVEILLANCE PROGRAM .............................................................................. 3 2.1 MEDICAL SURVEILLANCE .............................................................................................................................. 3 2.2 EMPLOYEE SERUM STORAGE ......................................................................................................................... 4 2.3 IMMUNIZATIONS ............................................................................................................................................. 4 2.4 MEDICAL RESTRICTIONS ................................................................................................................................ 5 2.5 EMERGENCY PROCEDURES FOR EXPOSURE INCIDENTS ............................................................................... 6 2.6 POST-EXPOSURE EVALUATION AND FOLLOW-UP ........................................................................................ 6 SECTION 3 LABORATORY PROCEDURES ................................................................................................ 7 3.1 ENTRY PROCEDURES ...................................................................................................................................... 7 3.2 EXIT PROCEDURES .......................................................................................................................................... 8 3.3 PERSONNEL PRACTICES .................................................................................................................................. 8 3.4 GENERAL LABORATORY PROCEDURES ......................................................................................................... 8 3.5 VACUUM LINE TRAPS AND FILTER SYSTEMS ............................................................................................ 10 3.6 BSL3 LABORATORY PRACTICES ................................................................................................................ 10 3.7 PERSONAL PROTECTIVE EQUIPMENT (PPE) ............................................................................................... 11 3.8 HANDWASHING ............................................................................................................................................ 12 3.9 PROCEDURES FOR CENTRIFUGATION .......................................................................................................... 12 3.10 BIOLOGICAL SAFETY CABINET (BSC) PROTOCOL .................................................................................... 13 3.11 CHEMICAL HOOD - GUIDELINES FOR OPERATION ..................................................................................... 15 3.12 WASTE HANDLING AND DISPOSAL PROCEDURES ..................................................................................... 15 3.13 EQUIPMENT REPAIRS/SERVICE ................................................................................................................... 15 3.14 PROCEDURE FOR LAUNDERING LABORATORY PROTECTIVE CLOTHING ................................................. 16 3.15 TRANSPORT OF HUMAN PATHOGENS OR OTHER POTENTIALLY INFECTIOUS MATERIALS ON CAMPUS (BETWEEN LABS OR BUILDINGS) ................................................................................................................ 16 SECTION 4 DECONTAMINATION OF SPILLS......................................................................................... 16 4.1 SPILLS IN BSC .............................................................................................................................................. 16 4.2 SPILLS IN THE LABORATORY ....................................................................................................................... 17 4.3 COMPOSITION OF BSL3 SPILL KIT .............................................................................................................. 17 SECTION 5 WORK WITH HAZARDOUS BIOLOGICAL TOXINS ...................................................... 19 5.1 RECOMBINANT DNA EXPERIMENTS INVOLVING TOXINS ........................................................................ 19 Yale University - Biological Safety BSL3 Laboratory Manual Revised September 2019 i 5.2 SELECT AGENT TOXINS ............................................................................................................................... 20 5.3 TOXIN WORK PRACTICES ............................................................................................................................ 20 SECTION 6 SELECT AGENTS ....................................................................................................................... 22 6.1 POSSESSION, USE, OR TRANSFER OF SELECT AGENTS .............................................................................. 22 6.2 LIST OF SELECT AGENTS AND REGULATED TOXINS ................................................................................. 23 6.3 TIER 1 SELECT AGENTS ............................................................................................................................... 24 6.4 PERMISSIBLE TOXIN AMOUNTS .................................................................................................................. 25 6.5 REPORTING SUSPECTED VIOLATIONS OR SUSPICIOUS ACTIVITY ............................................................. 25 6.6 REGISTRATION OF POSSESSION, USE OR TRANSFER OF SELECT AGENTS. ............................................... 26 6.7 DISCOVERY OF SELECT AGENTS OR UNKNOWN SAMPLES ....................................................................... 26 6.8 INTRAFACILITY TRANSFER OF SELECT AGENTS ........................................................................................ 27 6.9 DESTRUCTION OF SELECT AGENTS OR UNKNOWN SAMPLES ................................................................... 27 SECTION 7 PHYSICAL PLANT PERSONNEL, MAINTENANCE AND VISITOR PROCEDURES .. .......................................................................................................................................................... 28 7.1 VACCINATION REQUIREMENTS ................................................................................................................... 28 7.2 PROCEDURES FOR ENTERING BSL3 FACILITY ............................................................................................ 28 7.3 PERSONAL PROTECTIVE EQUIPMENT (PPE) ............................................................................................... 28 7.4 RESPONSIBILITIES OF THE BSL3 FACILITY USERS ..................................................................................... 28 7.5 RESPONSIBILITIES OF PHYSICAL PLANT PERSONNEL AND VISITORS ....................................................... 28 SECTION 8 REGISTERING YOUR WORK WITH THE EHS-BIOSAFETY OFFICE ...................... 29 8.1 REGISTRATION OF BIOLOGICAL MATERIALS ............................................................................................. 29 8.2 REQUEST TO USE INFECTIOUS AGENTS ...................................................................................................... 29 8.3 RECOMBINANT DNA REGISTRATION ......................................................................................................... 30 SECTION 9 AUTOCLAVE VALIDATION ................................................................................................... 30 9.1 BIOLOGICAL INDICATOR TESTS .................................................................................................................. 31 9.2 CHEMICAL TEST ........................................................................................................................................... 31 SECTION 10 FORMS ........................................................................................................................................... 31 10.1 INFECTIOUS AGENTS REGISTRATION ......................................................................................................... 32 10.2 REQUIREMENTS FOR BSL3 EXPERIMENTATION IN ANIMALS ................................................................... 32 10.3 REGISTRATION FOR BIOSAFETY LEVEL 3 (BSL3) RESEARCH IN ANIMALS ............................................. 34 10.4 SUPPLEMENTAL STANDARD OPERATING