1. Working Safely with Biological Materials
Basics of Biosafety
Working Safely with Biological Materials

2. What is Biosafety?
Principles and practices employed to protect laboratory personnel (& others in the area) and the 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 There is always risk! The risk must be identified
The “agent” is the what creates risk
Risks to the worker or environment are often unknown
Determining “acceptable risk”?
There is always risk!
The risk must be identified
The risk is evaluated
The risk must be measured
Plan to minimize the 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
We are working on this.

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: Mycobacterium tuberculosis, 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
“Medical waste” means any of the following that are not generated from a
household, a farm operation or other agricultural business, a home for the aged, or a
home health care agency: (a) Cultures and stocks of infectious agents and associated
biologicals, including laboratory waste, biological production wastes, discarded live and
attenuated vaccines, culture dishes, and related devices.
(b) Liquid human and animal waste, including blood and blood products and body
fluids, but not including urine or materials stained with blood or body fluids.
(c) Pathological waste.
(d) Sharps.
(e) Contaminated wastes from animals that have been exposed to agents infectious
to humans, these being primarily research animals.

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 Ethanol 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 Biohazardous Solid 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. Place sharps in approved sharps container for disposal!
Is it a sharps hazard? #3
Examples:
needles
syringes
scalpels
all biologically contaminated 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 containers must be used for both clinic and field 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 a one-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 mercury thermometers

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 days of 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?