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BIOSAFETY TRAINING

BIOSAFETY TRAINING. Office of Risk Management http://www.uottawa.ca/services/ehss/biosafety.htm. Lois Sowden-Plunkett Sept, 2 012. Challenge your understanding Establish good laboratory practices Prevent contamination (you and your work) Ensure research funding continues $$$.

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BIOSAFETY TRAINING

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  1. BIOSAFETYTRAINING Office of Risk Management http://www.uottawa.ca/services/ehss/biosafety.htm Lois Sowden-Plunkett Sept, 2012

  2. Challenge your understanding Establish good laboratory practices Prevent contamination (you and your work) Ensure research funding continues $$$ Because research is now interdisciplinary, it is now necessary to retool yourself with new skills and new understanding.

  3. Terminology: the source of confusion • BIOHAZARD • BIOSAFETY • BIOSECURITY • BIOSURETY

  4. What is a BIOHAZARD? A potentially infectious agent or hazardous biological material that presents a risk or potential risk to the health of humans, animals, plants, and the environment CFIA, PHAC, EC, DFAIT, TC, TRICOUNCIL & OTHER FUNDING SOURCES, NIH....

  5. viruses, bacteria, fungi, parasites, biological toxins, prions, and other micro-organisms or genetic systems, by virtue of their replicative properties, are potentially harmful to humans, animals, plants and/or the environment.

  6. Recombinant DNA, cultured cell lines, tissues and anatomical specimens from human or animal subjects, and blood and other bodily fluids are also potentially infectious unless tested and proven otherwise.

  7. found in your intestines • aid in food digestion • E.coli synthesize vitamins B1,B2 & K. • do not cause any harm if are in limited number. • deficiencies of these vitamins cause many diseases • beware of your antibiotics which can destroy your E.coli E. coli Extensively used for recombinant DNA application DH5 alpha BL 21 XL1Blue

  8. Risk may be strain dependent

  9. Pseudomonas spp. • P. aeruginosa • P. stutzeri • P. fluorescens • Ubiquitous in the environment • Opportunistic • affects humans, animals and plants • Survival out of host (months on dry surfaces) Risk groups Characteristics

  10. Bloodborne pathogens UPEI June 27, 2012 300 Students at potential risk • The testing involved a lancet and a receptacle for the blood. • The lancets were single use only, but the receptacles were used by several students. • Hepatitis B,C & HIV • Don’t assume because you know the person there is no risk • Don’t even assume you don’t have it. • If it’s not tested, it’s not safe! UNIVERSAL PRECAUTION

  11. What is BIOSAFETY? The combination of measures employed when handling biohazardous materials to: Protect personnel from exposure to infectious agents Prevent environmental release and contamination It’s good for you, it’s good for the science and it’s the law!

  12. How is biosafety achieved? Safety Principles: • Administrative controls • Engineering Controls • Practices and Procedures • Personal Protective Equipment In practical terms, this means: Training, risk assessments, authorizations, certification of rooms and equipment, appropriated practices, health assessment

  13. What is BIOSECURITY? Measures employed to protect biohazardous materials, or critical relevant information, against theft or diversion by those who intend to pursue intentional misuse. (selected agents, U99, BSE)

  14. How is biosecurity achieved? Physical barriers Buildings, doors, locks, key card access Psychological barriers Security personnel, cameras Monitoring Activities Patrols, monitoring by support staff Personnel Clearance Access to authorized personnel only

  15. What is BIOSURETY? Biosurety is defined as the combination of security, biosafety, agent accountability, and personnel reliability needed to prevent unauthorized access.

  16. To be safe and compliant it is really quite easy, it’s all about: Diligence Knowing who is responsible Knowing your risk Practicing GMP (good microbiological practice)

  17. Accountability Framework

  18. Impacted Stakeholders: Your faculty, your colleague, PS, PRS, HR ... Federal & Provincial Agencies Municipalities International Funding Agencies

  19. So let’s get a hand on this ! Pathogenic………………..Non-pathogenic Naturally occurring……..Manipulated Ubiquitous………………..Evolving/Rare

  20. CLASSIFICATION It’s as easy as: 1 -2-3-4 ! • Pathogencity • Infectious dose • Mode of transmission • Survival outside host and host range • Communicability • Immunization • Prophylaxis / Treatment

  21. WHMIS Class D, division 3 of WHMIS (Poisonous and Infectious Material - Biohazardous Infectious Material)

  22. RISK GROUPS:

  23. Recombinant DNA • In vitro incorporation of genetic material from one cell into another or from one organism to another • Level of risk depends on: • the source of DNA being transferred • the vector • the host Untransformed mammalian cell lines - Risk Group 1 • MCF-7 (Human breast carcinoma cell line) • NIH 3T3 (Mouse fibroblast cell line) Transformed mammalian cell lines – Risk Group 2 • HeLa (Human - contains papovavirus) Animal (may be infectious without your knowledge, or were intentionally injected with a pathogen Mammalian Cell Lines

  24. Endotoxins are part of the outer membrane of the cell wall of Gram-negative bacteria. Escherichia coli, Salmonella, Shigella, Pseudomonas, Neisseria, Haemophilus influenzae, Bordetella pertussis and Vibrio cholerae. Tetrodotoxin (TTX) is a potent neurotoxin with no known antidote. Includes unconventional agents, slow viruses and prions causing progressive neurological diseases ex CJD, BSE, Scrapie Resistant to destruction Precautions: • Handle tissues as Risk Group 2 or higher • Handle formalin-fixed tissues and paraffin-embedded blocks as if still infectious • Follow up-to-date disinfection protocols. PRIONS Toxins

  25. CONTAINMENT LEVEL: • measuresrequired for handling each organism safely in a laboratory setting • Specific to the risk group level and amounts being used • CL 1, 2, 3, or 4 ROOM INTEGRITY • Laboratory locations & access control • Air Handling and directional air flow • Work Surfaces • Agents Used • Lab services (water, drains, gas, electrical and safety) ( certification, commissioning)

  26. 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. • Petri dish, vial • Biological safety cabinets • animal caging equipment

  27. Secondary Containment • Protects the environment external to the laboratory from exposure • Includes facility design and operational practices Employs: Directional airflow Air and Drain filtration HEPA Filtration of lab air Pressure differentials Laboratory Design Operational Practices

  28. Freezer Fumehood Incubator Sink Coats 4 BSC Air intake office Air exhaust Dead air

  29. Containment Level 1 • Basic laboratory • Requires no special design features • Biosafety cabinets are not required and work may be performed on the open bench.

  30. Containment Level 2 • Clinical, diagnostic, research and teaching facilities with level 2 agents. • May require a class I or class II biological safety cabinet • Emergency plan • Access controlled

  31. Containment Level 3 • Specialized design and construction including commissioning and annual certification • Research projects reviewed by a specific panel • Standard operating procedures enforced for the safety of the individual and proper operation of the lab. • Personnel – additional training and supervision.

  32. Containment Level 4 Canadian Centre for Human and Animal Health in Winnipeg, Man. • Design specifications are extremely stringent • The worker is completely isolated from infectious material. • personnel security clearance and qualifications scrutinized

  33. Risk Assessment Why Bother LAI ???

  34. Laboratory Associated Infections Only 20% can identify the cause or event • 80% are caused by human errors • 20% are caused by equipment failure

  35. Types of accidents causing LAIs • Spills and sprays • Needles • Sharp objects and broken glass • Bites or scratches from animals Vaccinia virus Pseudomonas Attenuated – Lab Adapted Strains Laboratory-Acquired Infection With an Attenuated Yersiniapestis Strain—Chicago, Illinois, 2009

  36. BioRisk Assessment

  37. Agent characteristics & biological material • Is this a material you have used before? • Do you know the source and whether it has been tested for which agents and to prove it is non-replicating • What are the characteristics of the material upon receipt? • What are the implications of the manipulations you are planning? • Is this a material for which LAI have been reported or are materials of concern ( gov’t, society, etc)?

  38. SOURCE OF INFECTION • Microorganisms • Cells and tissues • Blood and body fluids • Any items contaminated with the above Laboratory Associated Infection • ROUTE OF TRANSMISSION • Percutaneous inoculations • Inhalation of aerosols • Contact of mucous membranes • Ingestion • SUSCEPTIBLE HOST • Immune system • Vaccination status • Age

  39. BLOODBORNE PATHOGENS (BBP) Semen Vaginal Secretions Tissue Cultures Organ Cultures Infected Experimental Animals Other Bodily Fluids: Cerebrospinal, Amniotic, Synovial In Blood & Other Potential Infectious Materials (OPIM)

  40. 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

  41. Internationally Recognized Resource Designed, Research And Maintained By PHAC And CFIA ONLY IN CANADA PATHOGEN MATERIAL SAFETY DATA SHEETS http://www.phac-aspc.gc.ca/lab-bio/res/psds-ftss/index-eng.php

  42. Personnel supervising & personnel using the material • your knowledge and experience • the level of mentorship available • New User Registration Form & Biosafety Health Assessment Forms • records your knowledge level • what you work with • health status Helps identify risk factors and potential for LAIs

  43. PPE – always your last defence • Criteria for consideration • Routes of exposure that need to be blocked • Degree of protection offered • Ease of use • Only effective if • correctly selected, fitted, used and cared for • the individual is well trained • Ensure PPE is removed before leaving the lab

  44. Gloves - yes they are mandatory • Double gloving a good practice • Gloves should not be reused • Gloves should be changed frequently • Glove selection: latex, nitrile, rubber & vinyl • Use the correct donning and doffing technique Herpes simplex virus (HSV)

  45. Lab Coats/Gowns Protect street clothing from spills Offer additional body protection Periodic cleaning required Echo virus type 9 Eyewear – need I say more ! NO CONTACTS ! Eye glasses, goggles, facemask Epstein-Barr Virus Infections

  46. Streptococci bacteria Footwear • Closed toe and heel shoes only. • No sandals! Facemask – prevent inhalation How many agents do you know that have flu-like symptoms. Influenza Virus

  47. UNIVERSAL PRECAUTIONS If samples cannot be guaranteed non-infective …… treat as infectious! Minimum standard of practice for preventing the transmission of BBP includes: • Education • Hand washing • Use safe work practice • Wearing appropriate protective equipment

  48. Environment: laboratory, facility, community Contain your biohazard (Primary and secondary containment) Regulatory RequirementS PHAC, CFIA, EC, Fed/Prov/Munipcial) Lab Design (conception, construction, renovations, maintenance) Control Access (Physical controls: lab and inventory)

  49. Experimental Protocols • have to be researched thoroughly • designed with safety in mind as well as research • engage the supervisor • protocols on-line: http://www.protocol-online.org • Remember once you start the protocol • you are in research mode, • so you better have thought of safety first! • Experimental Protocols & Lab Practices Equipment

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