Biosafety Training University of Ottawa *Office of Risk Management Human Resources - Occupational Health Disability & Leave v0501
Biosafety Outline • Introduction • Laboratory Associated Infections • Blood-borne Pathogens • Classification of Biohazards • Infection/Biohazard Control • Spill Response • Biomedical Waste • Regulations
What is Biosafety? • Measures employed when handling biohazardous materials to avoid infecting oneself, others or the environment. • Achieved through • Engineering Controls • Administrative Controls • Practices and Procedures • Personal Protective Equipment
What is a Biohazard? A potential hazard to humans, animals or the environment caused by a biological organism, or by material produced by such an organism Examples; • Viruses, bacteria, fungi, and parasites and their toxins. • Blood and body fluids, as well as tissues from humans and animals. • Transformed cell lines and certain types of nucleic acids .
Who’s Responsible, who are the Stakeholders? INTERNALLY • Vice-President (Research) • Committees • University Services (ORM, HR, PRS, PS)* • Deans, Chairs, Principal Investigators, Employees, Students • Manager of Biological Containment Suite EXTERNALLY • Health Canada • Canadian Food Inspection Agency • Transport Canada • Ontario Ministry of Labour • Emergency Response Personnel • Suppliers & Contractors • Community
Key Services • Office of Risk Management • Training • Interface with Regulatory Bodies • Biosafety Program • certifications • training • procedures • inspections • contingency planning • accident/incident follow-up
Key Services • HR (Occupational Health, Disability and Leave) • Medical surveillance • Immunizations • Medical Follow-up • Interface with Workplace Safety and Insurance Board
Why are we concerned about biohazardous materials? • Potential for acquiring a laboratory-associated infection (LAI) • Contamination of the environment • Contamination of research • Public perception*
Laboratory Associated Infections • Percutaneous inoculation • Inhalation of aerosols • Contact of mucous membranes • Ingestion Infection Source Susceptible Host • Immune system • Vaccination status • Age • Cultures and stocks • Research animals • Specimens • Items contaminated with above Route of Transmission
LAI’s • Only 20% causative or defined event • 80% of which are caused by human error • 20% are caused by equipment failure • Top 4 accidents resulting in infection • Spillages & splashes • Needle and syringe • Sharp object, broken glass • Bite or scratch from animals or ectoparasites http://www.weizmann.ac.il/safety/bio2.html
Bloodborne Pathogens (BBP) • Sources • Blood • Semen • Vaginal Secretions • Body Fluids • Cerebrospinal • Amniotic • Synovial • Tissue Cultures • Organ Cultures • Infected Experimental Animals
Risk of Exposure • Pathogen involved • Type of body fluid • Route of exposure • Duration of exposure • Volume of blood involved in exposure • Concentration of virus at time of exposure • PPE worn
Specific Examples of Bloodborne Pathogens Hepatitis B Hepatitis C HIV
Issues to Consider • Symptoms • Mode of transmission • Incubation period • Survival outside host • Communicability • Immunization • Prophylaxis / Treatment
If An Exposure Occurs (or the possibility of exposure) • Initiate first aid • Notify your supervisor / designated person • Report to hospital emergency department or University’s Health Services • Report incident to OHDL Occupational Health, Disability and Leave Office telephone ext. 1472 http://www.uottawa.ca/services/hr/frames.html
Universal Precautions • Minimum standard of practice for preventing the transmission of BBP Includes: - Education - Hand washing - Wearing protective barriers - Use safe work practices If samples cannot be guaranteed non-infective …… treat as infectious!
Classification of Biohazards • Conventional Agents • Unconventional Agents • Recombinant DNA • Tissue Culture • Animal Work • Anatomical Specimens Class D, division 3 of WHMIS (Poisonous and Infectious Material - Biohazardous Infectious Material)
Classification of Biohazards _ • As the level so does ; • the risk of the organism to humans, animals, plants and/or the environment • the procedural and facility requirements • the level of containment required • the degree of protection for personnel, the environment and the community. BSL 4 BSL 3 BSL 2 BSL 1 _
Conventional Agents Unlikely to cause disease in healthy workers or animals Rarely cause serious human or animal disease May cause serious disease Likely to cause very serious disease
Unconventional Pathogens • TSE prion diseases; lethal transmissible neurodegenerative conditions • Creutzfeld-Jakob disease, Variant C-J Disease, Mad Cow Disease, Scrapie, Chronic Wasting Disease. • Resistant to destruction by procedures that normally inactivate viruses. • Contact ORM to assess requirements (containment, procedures, waste disposal, etc.)
Recombinant DNA • Canada: Level of risk depends on source of DNA, vector and host. • The Biosafety Committee will assist the investigator in this determination. Genetic Engineering = in vitro incorporation of genetic material from one cell into another
Tissue Culture • Have the potential to contain pathogenic organisms • In general; Human & non-human primate, and mycoplasma-containing cell lines Level 2 Level 1 Others A detailed risk assessment should be undertaken when using a new cell line.
Animal Work • Animals can harbour infectious organisms (naturally or introduced) • Level dependent on type of work being conducted. • Special Animal Care training is required for all personnel working with animals. • All work involving animal use must receive prior approval from the Animal Care Committee
Anatomical Specimens • All specimens should be considered infectious due to potential presence of infectious agents • Important to consider the type of specimen • blood, organs, tissues • Spinal sample, brain tissue • From infectious patient • In general Level 2 but it depends on the nature of the work.
Infection/Biohazard Control Engineering Controls Administrative Controls Practices and Procedures Personal Protective Equipment
Engineering Controls • Technology based, reduce or eliminate exposure to hazards by changes at the source of the hazard. • Containment: • Primary vs Secondary • Containment levels
Primary Containment • First line of defence. • Ensures protection of personnel and immediate environment from exposure to the infectious agent. • ‘Protective envelope’ that encapsulates the infectious agent or animal. • Petrie dish, vial, stoppered bottle…. • Biological safety cabinets, glove boxes and animal caging equipment…. Effectiveness of control is based on the integrity of the containment.
Secondary Containment • Protects the environment external to the laboratory from exposure. • Includes facility design and operational practices.
Biosafety Containment Levels • Containment Levels similar to Risk Levels. • Biohazards Committee will evaluate the research proposals to ensure adequate containment . • Level 1 • Level 2 • Level 3 • Level 4
Level 1 • Basic laboratory • Requires no special design features • Biosafety cabinets are not required and work may be performed on the open bench.
Level 2 • Clinical, diagnostic, research and teaching facilities with level 2 agents. • Requires a class I or class II biological safety cabinet if any potential for aerosol or splash exists. • An emergency plan for handling spills must be developed. • Access should be controlled.
Level 3 • Specialized design and construction • primary barriers to protect the individual • secondary barriers to protect the environment • All staff must undergo special training on the agents being used, PPE, equipment, waste management as well as practices and procedures above and beyond the scope of this course.
Level 4 • Only one level 4 facility in Canada (Canadian Centre for Human and Animal Health in Winnipeg, Man.) • Design specifications are extremely stringent, worker is completely isolated from infectious material.
Biological Safety Cabinets • Effective means of physical containment for biological agents, especially when aerosols are generated. • HEPA filters remove particles (min 0.3 microns) with 99.97% efficiency. • There are 3 main classes of cabinets (I, II, III) which provide various levels of protection.
Biological Safety Cabinets VS • Laminar flow hoods • NOT biological safety cabinets • Vertical or horizontal laminar flow • HEPA filtered air (intake) • product protection only • Biological Safety Cabinet • HEPA filtered laminar air flow and • exhaust • personnel, environment & often • product protection
Working safely in a BSC Before using the cabinet: • Ensure BSC is certified • Turn off UV lamp; turn on fluorescent lamp • Disinfect work surfaces with appropriate disinfectant • Place essential items inside cabinet • Allow the blower to run for 5-10 min before work
Working safely in a BSC During use: • Ensure material and equipment is placed near the back of the hood, especially aerosol-generating equipment. Do not block any vents. • Use techniques that reduce splatter and aerosols. • General work flow should be from clean to contaminated areas. • Minimize movement so as not to impede air flow. • Open flame in BSC’s is controversial.
Working safely in a BSC After completion of work: • Leave blower on at least 5 minutes to purge cabinet • Remove and decontaminate equipment and materials • Disinfect cabinet surfaces • Turn off blower and fluorescent lamp, turn on UV lamp
Working safely in a BSC Maintenance: • Twice daily - Work surfaces wiped down • Weekly - UV lamp should be wiped clean* • Monthly - All vertical surfaces wiped down • Annually - UV lamp intensity verified. - Decontamination with formaldehyde gas (ORM) - Certification (ORM)
Administrative Controls Program based, information and methods to minimize risk of exposure. • Risk assessment • Medical Surveillance • Training/Education • Resources • Inspections • Signs & Labeling
Administrative Controls Risk Assessment • Will determine type of containment, procedures, and safety equipment required • Responsibility of users, additional assistance is available from ORM • Consider areas such as; experimental design, procedures to be employed and personal experience/knowledge, etc. * http://www.hc-sc.gc.ca/pphb-dgspsp/ols-bsl/lbg-ldmbl/pdf/lbg-3e-draft.pdf
Administrative Controls Risk Assessment: Know your Agent • Know and understand the various characteristics of the agent(s) you are working with. • This information is available from; • MSDS’s • Suppliers or manufacturers • Example
Administrative Controls Medical Surveillance Training & Education • Lab specific policies and procedures • Biosafety training Resources • ORM web site, Biosafety page • Faculty web sites • Biosafety Manual • Training Videos
Administrative Controls Inspections • Routine self-inspections • Biosafety Inspection Checklist available on-line • In addition, ORM, EHSOs and OH&S will inspect labs to ensure compliance with regulations/ guidelines and provide feedback.
Administrative Controls Signs & Labeling • Biohazard warning signs must be posted on doors to rooms where biohazardous materials are used. • Biohazard labels should be placed on containers, equipment and storage units used with biological agents.
Practices and Procedures • General Safety Guidelines • Good Microbiological Practice • Handwashing • Specific Procedures • Centrifuges • Needles & Syringes and other sharps • Pipettes • Blenders, Grinders, Sonicators & Lyophilizers • Inoculation Loops • Cryostats
General Laboratory Safety Guidelines • Mostly common sense, but you must understand the hazards you face in the laboratory and be adequately trained to deal with them. • Basic must knows for all labs. • Examples? b i o s a f e t y