Laboratory Biosafety Manual
(BSL Level)
Room (Number)
Building (Name)
Department (Name)
PI/Lab Director (Name)
Revised by OSEH 2/14/2013
Validation and History for Biosafety Manual
Director’s Certification:
I hereby certify that I have reviewed the contents of this manual and that it reflects my current operating policy for the laboratories (room numbers) located in (building).
(Principal Investigators Name – please print on line )
(Signature)
(Date)
(List all Co-Principal Investigators Name– please print)
(Annual Review Dates)
Chapter 1– Introduction 6
Acknowledgments 7
Definitions 7
Risk Group Classification 9
Laboratory Biosafety Level 9
Chapter 2 - General Information and Structure 10
Commitment to Safety 10
Management Structure and Responsibility 11
Oversight 11
Chemical Hygiene Plan (CHP) 11
Research Agents 11
Laboratory Access 13
Laboratory Security 13
Select Agents 13
Personnel Training 14
Facilities 15
Negative pressure tissue culture rooms 15
Bench Tops 15
Laboratory Furniture 15
Workplace Practice Controls 15
Vaccinations (if applicable) 16
Posting and Labeling Requirements 16
Signs 16
Labels and Tags 16
Audit Management 17
Storage of Biohazardous Materials 17
Transport of Hazardous Materials to Storage Areas 18
Chapter 3 - Laboratory Practices 18
Standard Microbiological Practices 18
Biosafety Procedures 18
Personal Protective Equipment 18
Laundry and Decontamination of Protective Clothing 19
Standard Microbiological Practices BSL2+ 20
Good Microbiological Techniques 21
Use of pipetting aids 21
Minimization of aerosol production 22
Protection of vacuum line 22
Syringes and Needles 22
Biological Safely Cabinet Use 23
Biohazardous Waste Disposal Methods 24
Biohazardous Glassware, Plastics and Other Solid Waste 24
Biohazardous Liquids– Sanitary Drain 25
OSEH Hazardous Materials Management (HMM) - Collection of Waste 25
Biohazardous Glassware, Plastics and Other Solid Waste 25
Biohazardous Regulated Sharps - Note Sharps cannot be treated by the laboratory 25
Biohazardous Liquids 26
Chapter 4 - Research Animals 27
Use of Sharps with Animals 28
Animal waste 28
Chapter 5 – Specific Standard Operating Procedures 29
Biological Safety Data Sheets 29
Chapter 6 – Equipment and Facility Management 30
Equipment Care and Use 30
Proper Use of Autoclave 30
Proper Use of Centrifuges 32
Spills and Leaks 33
Blenders, Ultrasonic Disrupters, Grinders and Lyophilizers 34
Miscellaneous equipment 34
Microscopes 34
Microtomes 34
Cryostats 34
Water baths 35
Loop Sterilizers and Bunsen Burners 35
Open Flames in Biosafety Cabinets 35
Other Physical and Chemical Hazards 36
Dry Ice 36
Liquid Nitrogen 37
Ultraviolet light 39
Electrical Hazards 40
Gel Electrophoresis 40
Ethidium Bromide 40
Acrylamide 40
Additional Chemical Usage 41
Phenol 41
Chloroform 41
Equipment Maintenance 42
Cleaning and Decontamination 42
Chapter 7 – Radiation Management 43
Use of Radioactive Isotopes 43
Disposal of Isotopes after Usage 44
Chapter 8 - Emergency Management 45
Emergency Procedures 45
Accidental Exposure to an Infectious Agent 45
Spill Response Procedures for Infectious agents and Recombinant DNA 46
Spill in the Laboratory 47
Skin, Mucous Membrane, or Injury Exposure 48
For exposures to recombinant DNA, perform the following: 48
Fire Fighting Procedures 49
Reporting and Recordkeeping 49
Chapter 9 - Shipment and Receipt of Infectious Materials 50
Receipt of Infectious Materials 50
Transportation of Biohazardous Materials 50
On-Campus Transportation 51
Hand Carry Between University Labs or Buildings 51
Transport by University Vehicle 51
Off-site Transportation by Courier 52
Chapter 10 – Appendices 53
Appendix 1 54
Emergency Call List 54
Appendix 2 55
Schematic of Laboratory 55
Appendix 3 56
Roster of Approved Personnel and Training Records 56
Appendix 4 57
Training Completion Form 57
Training Checklist 58
Appendix 5 61
Housekeeping Inspections 61
Appendix 6 62
Pathogen Safety Data Sheets for Infectious Agents 62
Appendix 7 63
Accident Report Forms 63
Appendix 8 65
Standard Operating Procedure 65
Appendix 9 68
Autoclave Use Log 68
Chapter 1– Introduction
While this manual is not intended to cover every aspect of biosafety, essential basic procedures, precautions and guidelines are discussed. If basic (BSL level) laboratory practices are performed carefully, along with the appropriate experimental design and common sense, workers should have few problems working safely in the (BSL level) environment.
It is essential that laboratory personnel maintain good, sound laboratory work practices and precautions. An awareness and respect for the dangerous nature of certain pathogens should always be maintained. Extensive guidelines regarding biosafety containment levels and various agent summary statements can be obtained from the U.S. Department of Health and Human Services, Centers for Disease Control and Prevention and National Institutes of Health publication entitled Biosafety in Microbiological and Biomedical Laboratories 5th Edition. This document is available from the Department of Occupational Safety Environmental Health (OSEH) website at http://www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm
The guidelines should be read before work in the (BSL Level) laboratory is initiated. In addition, those who plan to initiate work in the lab must be familiar with the standard practices that apply as well.
_____________________________ ___________________
Lab Director Date
Acknowledgments
This Biosafety Manual is the result of assessing the safety-management needs of a variety of laboratories that use biological materials at the University of Michigan. We would like to acknowledge and express our appreciation to all the researchers who contributed to this effort.
Definitions
Biohazardous Material - any material known to harbor organisms or agents capable of infecting or infesting human or animal hosts or causing environmental harm if released.
Biosafety - Biosafety is a specialized practice for proper handling and working with biohazardous organisms or biological material, which may harbor biohazardous organisms.
Biosafety fits neatly into the traditional programmed approach to safety:
· Administrative controls to standardize methods to reduce exposure risk
· Mechanical engineering controls for containment of hazardous materials
· Medical surveillance and PPE for potentially exposed employee
· Workplace monitoring to determine exposure levels
Biosafety Levels aka Biological Safety Level- (BSLs) describe the combination of safety practices, safety equipment and facility design used to contain the hazards associated with specific risk groups of microorganisms and is based on risk assessment. Biosafety levels are different from risk groups however risk group information is critical in determining the correct biosafety containment level. See below
Blood borne Pathogen - an agent known to be transmissible through contact with human blood, such as the human immunodeficiency virus (HIV) or the hepatitis B virus (HBV).
Containment:
Primary Containment – The protection of personnel and the immediate laboratory environment from exposure to infectious agents is provided by both good microbiological technique and use of appropriate safety equipment.
Secondary Containment – The protection of the environment external to the laboratory from exposure to infectious materials, is provided by a combination of facility design and operational practices
Infectious Substance - a viable micro-organism, or its toxin, which causes or may cause disease in humans or animals, and includes those agents listed in 42 CFR 72.3 or any other agent that causes or may cause severe, disabling, or fatal disease.
IBC - Institutional Biosafety Committee
Occupational Exposure – an exposure that may place personnel at risk of injury or infection is defined as percutaneous (e.g., a needle stick or cut with a sharp object), contact of mucous membranes, or contact of skin (especially when the exposed skin is chapped, abraded, or afflicted with dermatitis or the contact is prolonged or involving an extensive area) with blood, tissues, or other bodily fluids to which universal precautions apply. For the purpose of this manual, the occupational exposure must be to fluids or aerosols known to be infectious.
OSEH - University of Michigan, Department of Occupational Safety and Environmental Health
OVPR – Office of the Vice President for Research
Potentially Infectious Material - any material, which may or is known to contain an etiologic agent of human or animal disease.
Principal Investigator/Lab Director - the University of Michigan faculty member responsible for the research underway in the laboratory. For the purposes of this protocol, the Principal Investigator is (List PI, address and phone number)
Recombinant DNA Molecules - molecules that are constructed outside living cells by joining natural or synthetic DNA segments to DNA molecules that can replicate in a living cell; or molecules that result from the replication of those described above.
UCUCA - University Committee on Use and Care of Animals
Risk Groups- are the classification of infectious microorganisms based on principle characteristics such as; route of transmission and severity of disease. See below
Standard Microbiological Practices (SMP) - basic safe laboratory work protocols for working with biological materials based on containment level.
- The main objective of SMP is to provide safety controls needed to protect workers and the environment from contamination in the event that the agents are accidentally released from their primary container.
Risk Group Classification
Risk Group 1 (RG1) agents are defined and characterized strains of viable microorganisms not known to consistently cause disease in healthy adult humans or animals. Laboratory personnel may become infected through high doses or unusual routes of exposure that are not commonly encountered in a natural setting. Opportunistic RG1 pathogens may cause serious disease in elderly persons and infants, and persons with compromised immune systems. A risk assessment should be used for vaccine strains, as multiple passages in vivo do not ensure avirulence.
Risk Group 2 (RG2) agents are associated with human or animal disease which is rarely serious and for which preventive or therapeutic interventions are often available. RG2 organisms have the capability to cause serious disease based on dose, route of exposure, and immune status. Laboratory exposures may cause serious infection, but the risk of spread of infection is limited. The risk assessment should give special attention to those RG2 organisms for which preventative or therapeutic interventions are not available.
Risk Group 3 (RG3) agents are associated with serious or lethal human or animal disease and have the potential for respiratory transmission, and for which preventive or therapeutic interventions may be available. Preventative or therapeutic interventions may not be available for some RG3 agents, or are less available than for RG2 organisms.
Risk Group 4 (RG4) agents are associated with serious or lethal human or animal disease. RG4 agents may be transmitted via the aerosol route, can be readily transmitted from one individual to another, directly or indirectly, and for which there is no available vaccine or therapy. RG4 agents are NOT permitted at the University of Michigan.
Laboratory Biosafety Level
BSL1 practices, safety equipment, and facility design and construction include appropriate laboratories in which work is done with defined and characterized strains of viable microorganisms not known to consistently cause disease in healthy adult humans, and are of minimal potential hazard to laboratory personnel and the environment. The laboratory is not necessarily separated from the general traffic patterns in the building. Work is generally conducted on open bench tops using standard microbiological practices. Special containment equipment or facility design is neither required nor generally used. Laboratory personnel have specific training in the procedures conducted in the laboratory and are supervised by a scientist with general training in microbiology or a related science.
BSL2 practices, equipment, and facility design and construction are applicable to teaching and research laboratories in which work is done with the broad spectrum of indigenous moderate-risk agents that are present in the community and associated with human disease of varying severity. U of M requires all activities involving infectious materials be conducted in biological safety cabinets. No work with infectious materials in open vessels will be conducted outside the cabinets.
· RG2 agents infectious to humans or vertebrate animals are stored or manipulated;
· Human blood, blood products, or other potentially infectious materials, including commercial cell lines;
· animal tissues or fluids, especially those collected from nonhuman primates, that are contaminated with an agent infectious to humans or vertebrate animals, or materials that were collected from an animal known to be sick are manipulated, including commercial cell lines;
· Certain types of research utilizing toxins or venom.
BSL3 practices, safety equipment, and facility design and construction are
applicable to clinical, diagnostic, teaching, research, or production facilities in which
work is done with indigenous or exotic agents with a potential for respiratory transmission, and which may cause serious and potentially lethal infection.
Primary hazards to personnel working with these agents relate to autoinoculation, ingestion, and exposure to infectious aerosols. All laboratory manipulations should be performed in a BSC or other enclosed equipment, such as a gas-tight aerosol generation chamber. Secondary barriers for this level include controlled access to the laboratory and ventilation requirements that minimize the release of infectious aerosols from the laboratory.
· RG3 agents such as; Mycobacterium tuberculosis, St. Louis encephalitis virus, and Coxiella burnetii are representative of the microorganisms assigned to this level.
BSL-4 No work at BSL4 is conducted at the University of Michigan
Animal Biosafety Levels
A similar set of four biosafety levels are provided for work with vertebrate animals infected with agents which may infect humans. These Animal Biosafety Levels, ABSL 1 thru 4, provide for practices, equipment, and facilities that are comparable to the laboratory biosafety levels described above. However, there are unique hazards associated with infected animals that must be understood by those personnel with animal contact and addressed in the animal facility. Animal activity can create aerosols and bites and scratches can occur.
Chapter 2 - General Information and Structure
Commitment to Safety
It is the objective of this laboratory, and its management to practice safety in science and to exercise all reasonable and prudent precautions generally accepted as research industry standards. Guidelines recommended by the CDC and NIH for biosafety will be strictly observed and enforced by the laboratory director and management. At the conclusion of initial personnel training, each lab employee will sign a Training Completion Form (See Appendix 4) to be retained with the training records.
Management Structure and Responsibility
Overall supervision of the (BSL level) laboratory is the responsibility of (Name of Principal Investigator). Management of daily operations in the lab is the responsibility of (Name of Lab Manager or Responsible Person). Access to the laboratory must be obtained from one or both of the above.
Oversight
This plan meets the quality standards for laboratory safety management as required by UM-OSEH and OVPR.
Chemical Hygiene Plan (CHP)
The Laboratory Safety Standard requires employers to write and implement laboratory-specific Chemical Hygiene Plans (CHP). According to this regulation, a Chemical Hygiene Plan applies to all employers engaged in the laboratory use of hazardous chemicals. A “laboratory” is defined as a facility where the laboratory use of hazardous chemicals occurs. A “hazardous chemical” is defined as a chemical for which there is evidence that acute or chronic health effects may occur in exposed employees. The CHP should include specific work practices, procedures, and policies to ensure that employees are protected from all potentially hazardous chemicals used. Recommendations for completing each section of the Chemical Hygiene Plan are provided on OSEH’s Internet web site. The web site contains a generic CHP that can be down loaded and modified for use in your Lab (http://www.oseh.umich.edu/research/chem-hygiene.shtml ). If you have any questions regarding the CHP, please contact OSEH at 76 3-6973.