Basics of Biosafety

1. Basics of Biosafety Working Safely with Biological Materials

2. What is Biosafety? • Principles and practices employed to protect laboratory personnel (& others in the area) andthe environment from exposure or infection while working with living organisms, biological materials, or agents. • Included are any materials that may be potentially infectious. • Includes recombinant DNA research

3. Agents and Risks • The “agent” is the what creates risk • Risks to the worker or environment are often unknown • Determining “acceptable risk”?

4. Who Determines Acceptable Risk? • Assessment is conducted by a Biosafety Professional in partnership with and based on information provided by the Principal Investigator • The assessment is presented to the Institutional Biosafety Committee (IBC) for approval

5. Identifying Risk • Understand the biology of the agent • Susceptibility and transmission within the host • Hazards associated with equipment and procedures • Goal: • Provide the highest practical protection and the lowest practical exposure

6. Evaluating Risk Acceptability • Worst case scenario -What might happen? • Likelihood of an event • Seriousness of the incident • Actions needed to resolve the problems

7. WHO-World Health Organization Agents Assigned Risk Groups • RG-1 Unlikely to cause disease in humans or animals • low individual or community risk • RG-2 May cause disease but typically not serious • individual risk, low community risk, treatable • RG-3 May cause serious disease, usually treatable • High individual but low community risk, serious respiratory agents • RG-4 Serious or fatal, often not treatable, • Easy transmission, high individual and community risk

8. Biosafety Levels (BSL) • Different than the Risk Groups!! • Risk groups used in risk assessment • BSL are used in risk management • BSL are ways to control the agent • facilities, safety equipment, practices, PPE, etc. • Once risk is assessed then the appropriate BSL is determined

9. BioSafety Level 1 • Well characterized, non-pathogenic organisms or agents • Open bench- no containment • Use good laboratory practices, waste disposal, and aseptic techniques • Example: E. coli K-12 strains

10. BioSafety Level 2 • Agents of moderate hazard to personnel or environment • Basic lab, but restricted access, containment during certain processes (i.e. aerosols, large volumes, etc.) • Autoclave and Biological Safety Cabinet desired • Use good laboratory practices, waste disposal, and aseptic techniques • Example: most non-respiratory, non lethal, agents

11. BioSafety Level 3 • Agents of high hazard to personnel or environment • Respiratory exotic or indigenous agents which are easily transmissible causing serious or lethal disease • All work is contained, engineering controls and controlled environments we currently do not have the facilities to handle. Example: Mycobacteriumtuberculosis, SARS, etc.

12. BioSafety Level 4 • FORGET ABOUT IT!!! • Hemorrhagic fever, deadly viruses, etc. • Total containment, airtight labs, “submarine” doors, air pumps, water treatment, HEPA filtration, etc. • Positive pressure “moonsuits”

13. Laboratory Acquired Infections (LAI) Bacterial: 76% from clinical labs 8% from research labs Exposure: 60% acquired from inhalation Other exposures include: digestion, sharps, splashes, direct and indirect contact

14. Laboratory Acquired Infections (LAI) Viral • 16% from clinical labs • 70% from research labs • 32% from animal related activities

15. Biohazardous/Medical Waste • Waste that is potentially infectious to humans, animals or plants. It includes: • Medical Waste according to MMWRA • Regulated Waste by MIOSHA • Regulated Waste by CDC/NIH

16. Michigan Medical Waste Regulatory Act (MMWRA) • Defines “medical waste” • Requirements for waste handling and disposal • Requires generators to register with DEQ and implement a Medical Waste Management Plan

17. Biohazardous Waste Management Plan Must outline how generating facility complies with the MMWRA: • Types of wastes generated • Storage and disposal of wastes • Contingency plans • Training

18. Biohazardous Waste Categories a) Cultures and stocks of infectious agents and associated biologicals • laboratory waste • biological production waste • discarded live and attenuated vaccines • culture dishes and related materials • contaminated PPE

19. Biohazardous Waste Categories b) Liquid human and animal waste • liquid or semi-liquid blood and blood products and body fluids • contaminated items that would release blood or items that are caked with blood or other potentially infectious materials; NOT including urine or materials stained with blood or body fluids • infectious animal waste (research)

20. Biohazardous Waste Categories c) Pathological waste • tissues • body parts other than teeth • products of conception • fluids removed by trauma or during surgery or autopsy/necropsy or other medical procedure and not chemically fixed.

21. …And More Biohazardous Waste Categories • Animal and plant pathogen waste • Recombinant DNA waste • Sharps

22. Biowaste vs. Trash 3 basic questions to differentiate: • Is it contaminated with viable biological material or recombinant DNA? • Can blood or other regulated body or biological fluids be released? • Is it a sharps hazard? If the answer to any of these questions is yes, then you have a biowaste.

23. Is it contaminated with viable biological material or recombinant DNA? #1 • Examples: • Contaminated lab waste • Personal protective equipment used for handling potentially infectious materials (including handling infected animals or their products) • Wastes from infectious disease research (carcasses, body fluids…)

24. Can blood or other (regulated) body fluids or viable biological materials be released? #2 Some Examples… Tubes of blood Vacuum flasks containing body fluids or cell line waste

25. Managing Liquid Biohazardous Waste Storage: • Label and secure bulk vessels if not disposed of immediately Treatment: • Chemical disinfection OR • Autoclave Disposal:THEN • Flush to sewer • Use proper PPE!

26. Disinfection • 10% bleach solution • good for general disinfection • High organics use 20% • Needs to be made weekly • Test contact time* • Ethanol • Use 70% solution (most effective) • Longer contact time and flammable *Researchers should investigate and know effectiveness and contact time for the best disinfectant against your agent!

28. Managing Non-Sharp BiohazardousSolid Waste • labeled container • lined with a biohazardous waste bag • equipped with a lid.

29. The Autoclave or Steam Sterilizer

30. Moist Heat Sterilization • Proper autoclaving is important • All air must be flushed out of the chamber • The chamber should not be overpacked with solids • Large volume of liquids may take longer to autoclave due to the high specific heat of water. • To determine if the proper conditions have been met, we use indicators • Tape – indirect indicator • Biological indicator – paper strip covered with Geobacillus spores

31. Effective Waste Autoclaving • Leave bag open during autoclaving or loosely closed • Add water to bag prior to autoclaving if primarily dry materials • Steam must contact materials • Place bag in autoclavable tray with sides

32. Treated Waste Bag Disposal • Allow waste bag to cool • Use fume hood to reduce odors • Securely tie bag shut • Place bag in a non-transparent black bag for regular disposal Remember: NO ORANGE BAGS IN DUMPSTER!

33. Is it a sharps hazard? #3 • Examples: • needles • syringes • scalpels • all biologicallycontaminated objects that can easily penetrate skin (Pasteur pipettes,razor blades, etc.) Place sharps in approved sharps container for disposal!

34. …Syringes in research settings should be disposed of as a sharp to avoid public relations concerns!

35. Sharps Containers • Containers must be leak-proof, puncture-resistant, closable & labeled with the biohazard symbol. • Proper sharps containersmust be used forboth clinic andfield work.

36. Proper Use of Sharps Containers • Place tops on containers before use on lab bench • Don’t forget to date the container when first put into use • Remember: sharps containers are aone-way disposal system

37. Proper Use of Sharps Containers Use sharps containers for sharps ONLY! • No solid biohazardous waste (i.e. gauze, un-broken pipettes, gloves) • No mercurythermometers

38. What’s wrong with this picture?

39. Sharps Container Disposal • Containers must be permanently closed and disposed of via our hazardous waste company • Within 90 daysof first use • When ¾ full • Disposal methods: • Landfill • Incineration • We use waste hauler

40. Safety Notes on Sharps Use • Do not re-cap sharps • Keep sharps container in close proximity to point of use (i.e. limit handling) for easy disposal • Do not leave needles in pockets of coveralls or smocks

41. Carcasses and Body Parts • Human tissues • Unfixed tissues are medical waste • Make waste unrecognizable! • Animal tissues, carcasses • When generated in infectious disease or recombinant DNA research, these are medical waste • These items must be stored in biolabeled, leakproof containers for incineration.

42. Managing All That Other Waste… • Drain bottles of non-hazardous materials before disposal in trash • <3% of volume is considered empty • Higher volumes must not be thrown in the trash

43. Managing All That Other Waste… Do NOT discard medications in the trash. Treat as chemical waste.

44. Any Questions?