Purdue University Biological Safety Review

1. Purdue University Biological Safety Review Radiological and Environmental Management (Environmental Health Section)

2. Institutional Biosafety Committee The University’s Institutional Biosafety Committee (IBC) is the campus-based committee that has the responsibility for reviewing and approving all proposals, activities, and experiments involving recombinant DNA, biohazardous materials, and unfixed Human tissues, cell lines, or fluids.The IBC reviews processed protocol applications that deal with Class II or higher biohazards, unfixed human blood or tissues, or recombinant or synthetic DNA materials. Principal Investigators must complete an IBC protocol application (Form 1A).

3. IBC Procedures • Form 1A covers the basic project information, a facility inspection, a listing of personnel who will be involved, and assures that bio-awareness training is provided. • The Biosafety Officer then conducts a lab or facility inspection and assigns a biosafety level. The IBC Chair completes the process by reviewing the project procedure and either approves, disapproves, or exempts the protocol.

4. National Institutes of Health (NIH) Biohazard Risk Groups • Class 1: Agents of no or minimal hazard under ordinary conditions or handling. • Class 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. • Class 3: Includes pathogens that require special conditions for containment. • Class 4: Require the most stringent conditions for their containment because they are extremely hazardous to laboratory personnel or may cause serious epidemic disease.

5. Biohazard Exposure BIOHAZARDS: • Any organism/biologic by-product/allergen that causes ill effects to people, plants, or animals. • They can be either a fungus, bacteria, virus, prion, parasite, toxin, or DNA component. • They can be human body fluids, unfixed tissues, cell lines, or genetic oncogenes. EXPOSURE to biohazards may occur from: • Puncture wounds • Contact with abraded skin • Absorption through respiratory tract, oral route, or mucous membranes Principal Investigators who are unsure if a material is a biohazard should contact the University Biosafety Officer.

6. Biohazard Classifications Biosafety levels corresponds to the Class rating of the biohazard(e.g. BSL2 is required for Class II biohazards). BSL2 is the most common type of biohazard project that occurs on campus. Labs working under these conditions must have the following procedures in place: • Limited access • Door posts warnings • Surface decontamination methods • Waste handling • No eating or drinking • Hand washing • Appropriate personal protective equipment • A Class 2 Biosafety Cabinet (if aerosol production is anticipated).

7. Bio-Awareness Training Topics • Specific handling techniques for each class II 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. • Special hazards (e.g. complications for pregnancies and immuno-compromised lab staff) must be explained and posted. • Appropriate decontamination procedures must be reviewed. • Any required security steps must be taken.

8. Other Review Topics Personal Protective Equipment (PPE)…........ Waste Handling………………………………… Biological Safety Cabinets (BSC) &Laminar Flow Clean Benches (LFCB)……….. Biohazard Spills………………………………... Security……………………………………......... Slides 9-13 Slides 14-22 Slides 23-26 Slides 27-31 Slides 32-34

9. Personal Protective Equipment (PPE)

10. PPE Includes the PROPER • Goggles • Gloves • Footwear • Lab Coat • Respiratory Protection For the Task

11. PPE – What You Need to Know • The last barrier of defense between the user and the hazard. • Must be available for every body part as well as for respiratory protection. • Proper selection is critical for each individual. • Know how to don and doff each piece of PPE. A typical selection of PPE for biohazard application would be the use of nitrile or latex gloves, goggles, lab coat, closed toe shoes, and perhaps a N-95 dust and vapor mask.

12. PPE – Respiratory Protection • Respiratory protection comes in many varieties. • Choose the appropriate dust and vapor mask or respirator for the task.

13. PPE – Dust and Vapor Masks • Dust and vapor mask such as N-95 and N-100, are designed to trap minute particles and minor aerosols. These are recommended for use when handling biosafety level 2 agents outside of a biosafety cabinet or if there are aerosols produced in environments such as animal BSL2 facilities. • There are many different styles of dust and vapor masks available making it important to find a mask that will provide optimum facial fit and comfort.

14. Waste Handling

15. Waste – 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. • Dispose of in properly labeled puncture proof containers and never discard in the regular trash. • Call REM for pickup.

16. Waste – Sharps (2) DO NOT PLACE SHARPS CONTAINERS IN THE REGULAR TRASH!

17. Waste – Sharps (3) ALWAYSplace sharps in the appropriate sized puncture resistant container for disposal.

18. Waste – Sharps (4) BE CONSIDERATE • Laboratory employees that routinely work with sharps and building services personnel who handle wastes are at risk of being punctured or lacerated during their workday. • Report sharp related injuries to your supervisor and to the Biosafety Officer immediately.

19. Waste – 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.

20. Waste – Biohazard Bags (2) ALWAYS REMEMBER When handling a biohazard waste bag, be careful not to press the bag against your body. This prevents improperly disposed of sharps accidents. Do not create aerosols by forcing the air out of the waste bag.

21. Waste – Biohazard Bags (3) Protect yourself from the contents: • Hold Bio bags away from your body. • Don’t squeeze the bag so that aerosols are released into your breathing space. • Take biowaste to the autoclave room as soon as possible.

22. Waste – 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 protection equipment (e.g. heat-resistant gloves, lab coat, and face shield). • Do not put the following items in the autoclave: • Sealed containers • Aerosol cans • Volatiles • Uncontained sharps

23. Biological Safety Cabinets (BSC)& Laminar Flow Clean Benches (LFCB)

24. BSC vs. LFCB Biological Safety Cabinet Laminar Flow Clean Bench 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 • Protects the operator • Protects the environment • Protects the product • Reduces the risk of airborne infection by reducing the escape of aerosols

25. Biological Safety Cabinets • Biological safety cabinet usage must be combined with good work practices for optimum safety and contamination control. • Movement of arms into and out of the cabinet can disrupt airflow, adversely affecting the cabinet’s performance. Avoid bringing non-essential supplies and equipment into the cabinet. • Place supplies, equipment and absorbent towels so that air intake or exhaust grilles are not obstructed.

26. Biological Safety Cabinets (2) • Keep opening and closing of lab doors and other personnel activity to a minimum. • If a burner is deemed to be indispensable use an on demand type. • Work at least 4-6 inches inside the cabinet window. • Decontaminate spills as soon as they occur; remove and disinfect the grille if contaminated. For more instructions see the Biological Safety Manual.

27. Biohazard Spills

28. Biohazard Spills • Evacuate the lab for thirty minutes if the spill is a large volume of biohazard material. This allows aerosols to settle. • Lab staff decontaminating small biohazard spills need to have specific bio-agent awareness training, and plan in advance for cleanup of emergency spills. • If spill occurs inside a biological safety cabinet, leave cabinet turned on and decontaminate in place. • Use the proper personal protective equipment (e.g. impervious gloves, outerwear, goggles, etc.)

29. Biohazard Spills (2) • Spray the contaminated surfaces with a 1-10 solution of bleach and water. Be careful not to contaminate the outside of the spray bottle. • Remove all traces of the spill with paper towels or other acceptable materials and re-spray the cleaned area with the bleach solution and allow the air to dry. • Remove all traces of the spill with paper towels or other acceptable materials and re-spray the cleaned area with the bleach solution and allow the air to dry. REPORT ALL SPILLS TO YOUR SUPERVISOR AND THE BIOSAFETY OFFICER

30. Biohazard Spills (3) In the event of a spill: • Use the proper PPE • Use the proper disinfectant solution. • Dispose of clean-up materials in biohazard bags.

31. Commonly Used Disinfectants • Alcohols:ethyl or isopropyl alcohol at 70-80 percent concentration is a good general purpose disinfectant; not effective against bacterial spores. • Phenols:Effective against gram-positive bacteria, fungi, and lipid-containing viruses. • Quaternary Ammonium Compounds:Cationic detergents which are strongly surface active; extremely effective against lipophilic viruses; not effective against bacterial spores. • Chlorine:Low concentration (50-500 ppm) active against vegetative bacteria and most viruses; higher concentration (2500 ppm) required for bacterial spores; corrosive to metal surfaces; must be prepared fresh; laundry bleach (5.25 percent chlorine) may be diluted and used as a disinfectant. • Iodine:Recommended for general use; effective against vegetative bacteria and viruses; poor activity against bacterial spores.  Many of these disinfecting agents can be irritating and toxic and should be used in accordance with label direction for personal protective equipment, concentration, and contact time.

32. Security

33. Security Responsibilities • EVERYONE is responsible for security in the laboratory. • If you are using Class II biohazards make sure these materials are secured. • Keep your lab doors locked as well as the doors to areas outside your lab where Class II 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 Department Head and to the University Biosafety Officer.

34. Select Agent Security Special security measures apply to users of Select Agents. See the Biosafety Officer for more specific details. Ω