Biological Safety Training

1. Biological Safety Training modified slightly from REM 2008 Rachael DeRudder

2. Biohazards BIOHAZARDS: • A biological hazard or biohazard is a self-replicating organism, or substance derived from an organism, that poses a threat to (primarily) human health. This can include medical waste or samples of a microorganism, virus or toxin (from a biological source) that can impact human health. It can also include substances harmful to animals and plants. • Human body fluids, unfixed tissues, cell lines, or genetic oncogenes • Can also be a fungus, bacteria, prion, parasite, or DNA component.

3. Exposure EXPOSURE to biohazards can occur by • Injection (contact with open wound or abraded skin) • Inhalation (absorption through respiratory tract • Ingestion (e.g. contam.fingers in mouth) • Absorption through mucous membranes (e.g. contam. finger in eye or nose) Intact skin is regarded as an effective barrier.

4. NIH Biohazard Risk Groups • RG 1: Agents of no or minimal hazard under ordinary conditions or handling • RG 2: Includes agents which may produce disease of varying degrees of severity from accidental inoculation or injection or other means of cutaneous penetration but which are contained by ordinary laboratory techniques.

5. NIH Biohazard Risk Groups • RG 3: Includes pathogens that require special conditions for containment. • RG 4: Require the most stringent conditions for their containment because they are extremely hazardous to laboratory personnel or may cause serious epidemic disease.

6. Biohazard Classifications Biosafety levels corresponds to the rating of the biohazard, i.e., BSL2 is required for Risk Group (RG) 2 biohazards. BSL2 is the most common type of biohazard project that occurs on campus.

7. BSL2 • Limited access, • Door posts warnings, • Surface decontamination methods, • Waste handling, • No eating or drinking, • Hand washing, • Appropriate personal protective equipment, and • A Class 2 Biosafety Cabinet (if aerosol production is anticipated).

8. Bio-Awareness Training • Specific handling techniques for each RG 2 or higher pathogen that is used must be reviewed and explained by the Principal Investigator. • Signs and symptoms for the onset of disease caused by the organism must be reviewed.

9. Bio-Awareness Training • Special hazards, i.e., complications for pregnancies and immuno-compromised lab staff, must be explained and posted. • Know the proper Personal Protective Equipment (PPE). • Appropriate decontamination procedures must be reviewed. • Any required security steps must be taken.

10. PPE Personal Protective Equipment

11. PPE can include the PROPER • Goggles • Gloves • Footwear • Lab Coats • For the Task

12. Respiratory Protection comes in many varieties. Choose the appropriate mask for the task.

13. Waste Handling

14. Waste Handling Biohazard Bags BIO-RELATED WASTE MATERIALS: • Dispose of in properly labeled water impervious autoclavable bags (usually red or orange bags). • Do not over fill bags. • Utilize proper personal protective equipment and wash hands after completion. • Keep bags closed until they are ready to be decontaminated. • Do not dispose of a bio waste bag in the regular trash. • Do not place these bags in public areas such as hallways. Contact REM for removal of deconned and autoclaved materials.

15. Protect yourself from the contents. Hold Bio bags away from your body. Don’t squeeze the bag so aerosols are released into your breathing space. Take biowaste to the autoclave room as soon as possible.

16. Waste Handling - Autoclave • Autoclaves are very effective decontamination devices that use high pressure and steam (270F) to eliminate biological activity on equipment, wastes, or growth mediums. • Extreme heat and high pressure produced in autoclaves can cause serious injury. Always read the operations manual and review usage procedures with the lab manager. Users should understand and respect the hazards these can create and use the proper personal protective equipment, i.e., heat-resistant gloves, lab coat, and face shield. • Do not put the following items in the autoclave: - Sealed containers - Aerosol cans - Volatiles - Uncontained sharps

17. Waste Handling - SHARPS • SHARPS are items such as glass and plastic pipettes, broken glass, test tubes, petri dishes, razor blades, needles, syringes, etc. that are capable of puncturing, cutting, or abrading the skin. • Properly handle, decontaminate, and dispose of sharps in order to prevent injury and potential disease transmission.

18. DO NOT PLACE SHARPS CONTAINERS IN THE REGULAR TRASH! CALL REM FOR PICKUP

19. ALWAYS place sharps in puncture resistant containers for disposal.

20. In the event of a spill: Use the proper PPE Use the proper disinfectant solution. Dispose of clean-up materials in biohazard bags.

21. Security • EVERYONE is responsible for security in the laboratory. • If you are using RG 2 biohazards make sure these materials are secured. • Keep your lab doors locked as well as the doors to areas outside your lab where RG 2 organisms are incubated, stored, or otherwise processed. • Ask for identification from unknown individuals who enter your lab. Do not allow access to unauthorized people. • Secure biohazardous waste and autoclave as soon as possible. • Report the disappearance of any biohazardous material to your Supervisor. Special security measures apply to users of Select Agents. See the Biosafety Officer for more specific details.

22. HEPA Filtered Cabinets Biological Safety Cabinet Laminar Flow Clean Bench HEPA stands for: High Efficiency Particulate Air

23. Biological Safety Cabinets Laminar Flow Clean Benches • Protects operator • Protects the environment • Protects the product • Reduces the risk of airborne infection by reducing the escape of aerosols • Protects the product • Blows aerosols into the operator’s face. • Does not protect the operator • Should not be used with potential pathogens • Is not a Biological Safety Cabinet

24. BIOSAFETY CABINET

25. BSC – Safe for Biohazard Use Visual Indicator Sash

26. BIOSAFETY CABINET AIRFLOW (Courtesy of Eagleson Institute)

27. BIOSAFETY CABINETS • Class II Biosafety Cabinets, or BSCs, use filtered air to isolate materials from the operator and work environment. • BSCs use multiple filtration systems, fans, and air-flow paths that must be carefully controlled, maintained and checked to ensure operator safety. Most BSCs will not be vented to the outside. • BSCs protect both sample and workers from contaminating each other…designed to contain biological hazards (particles) not chemical fumes & vapors. • There are other Classes of Biosafety Cabinets. Not to be confused with Clean Bench or Chemical Fume Hood.

28. Working Safely in a BSC • Turn the blower on at least 5 minutes BEFORE beginning work to allow the BSC to remove any particulates in the cabinet. • Check the Magnehelic gauge before using every time . Reading should be equal to approximately 0.5 inches to assure proper operation of the cabinet before placing any materials into it. Higher readings may indicate filter clogging. Zero readings may indicate loss of filter integrity.

29. Working Safely in a BSC • Use the APPROPRIATE Personal Protective Equipment (PPE) while working at the BSC. • Place necessary materials in the BSC BEFORE beginning work to minimize the number of arm movement disruptions to the air barrier of the cabinet. • When working in a BSC, work with arms raised slightly above the front grille, NOT resting flatly on it. This alleviates the problem of room air flowing directly into the work area.

30. Working Safely in a BSC • DON’T place objects directly on the grillwork at the opening or back of the cabinet. This greatly interferes with the laminar airflow curtain. • After placing your equipment inside the cabinet, close the sash to the proper operating height. • AVOID working in and operating the cabinet with the sash in any other position than the manufacturer’s recommended sash height.

31. Working Safely in a BSC • The worker’s face should be ABOVE the front opening of the BSC. • All work should be performed at least 4inches from the front grille on the work surface.

32. Completing Work in the BSC • All equipment that has come in contact with a biological agent should be decontaminated.  • The cabinet should be allowed to run for at least 3 minutes with no activity.

33. Completing Work in the BSC • Wipe down all surfaces with the appropriate disinfectant, including the cabinet sides, back, and the interior of the glass. • Decontaminate or, where appropriate, autoclave all disposable materials and wastes before removal from the laboratory. • Call REM for pickup.

34. More Errors to Avoid • NEVER disengage the alarm. Alarm indicates improper airflow that could effect performance and endanger the researcher or the experiment. • Never completely close the window sash with the motor running.  This could damage the motor.

35. CLEAN BENCH UNSAFE for Biohazard Use

36. CLEAN BENCH AIRFLOW POTENTIALLY CONTAMINATED AIR HEPA FILTERED AIR ROOM AIR Courtesy of Eagleson Institute

37. CLEAN BENCH • Clean Benches: typically provide only product protection by creating a unidirectional airflow through a HEPA filter. • The discharged air goes directly into the workroom. • These cabinets are not safe for work with biohazards, chemicals, or radioactivity.

38. REMEMBER • A Biosafety Cabinet is only as safe as the person using it. • Use the APPROPRIATE cabinet type for the materials being used. • Make sure the cabinet is working properly BEFORE beginning work. • Consult the supervisor about questions or concerns regarding working in a Biosafety Cabinet.

39. Select Agents • Agents that could be used for terrorist acts • Highly regulated • Stiff penalties • FBI background checks • High security No one at Purdue has been approved for Select Agents at this time.

40. Resources Center For Disease Control and Prevention http://www.cdc.gov/ Select Agent Program http://www.cdc.gov/od/sap/ Biosafety in Microbiological and Biomedical Laboratories 5th Edition, January 20007 http://www.cdc.gov/od/ohs/biosfty/bmbl5/BMBL_5th_Edition.pdf/

41. Resourcescontinued Material Safety Data Sheets For Research Pathogens http://www.phac-aspc.gc.ca/msds-ftss/index.html Purdue University Biosafety Manual http://www.purdue.edu/rem/home/booklets/bioman.pdf Guidelines for Research Involving Recombinant DNA Molecules (NIH Guidelines) http://www4.od.nih.gov/oba/rac/guidelines/guidelines.html