Biological Safety Training

1. Biological Safety Training Radiological and Environmental Management Environmental Health Purdue University October 2003

2. Institutional Biosafety Committee (IBC) Procedures • 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 DNA materials. Principal Investigators must complete an IBC protocol application (Form 1A).

3. Institutional Biosafety Committee (IBC) Procedurescontinued • 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. Go to: IBComm@purdue.edu to contact the committee.

4. 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. • Can be either a fungus, bacteria, virus, prion, parasite, toxin, or DNA component. • 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, ibcomm@purdue.edu

6. Biohazard Classifications Biosafety levels corresponds to the Class rating of the biohazard, i.e., 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, and • A Class 2 Biosafety Cabinet (if aerosol production is anticipated).

7. Bio-Awareness Training • 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, i.e., 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. Personal Protective Equipment (PPE) PPE: • The last barrier of defense between the user and the hazard. • Must be available for every body part as well as 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.

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

10. 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.

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

12. 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.

13. Waste Handling - Biohazard Bagscontinued ALWAYS REMEMBER When handling a bio waste bag, be careful not to press the bag against your body. This prevents improperly disposed of sharp accidents. Do not create aerosols by forcing the air out of the waste bag.

14. 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.

15. 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 protection 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

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

17. DO NOT PLACE SHARPS CONTAINERS IN THE REGULAR TRASH!

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

19. Waste Handling – SHARPScontinued 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.

20. 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

21. Biosafety Cabinet Procedures • 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 cabinet performance. Avoid bringing non-essential equipment and supplies into the cabinet. • Place supplies, equipment and absorbent towels so that air intake or exhaust grilles are not obstructed.

22. Biosafety Cabinet Procedurescontinued • 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 Purdue’s Biological Safety Manual

23. 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 Biosafety Cabinet, leave cabinet turned on and decontaminate in place. • Use the proper personal protective equipment (PPE), e.g. latex gloves, outerwear, goggles, etc.

24. Biohazard Spillscontinued • 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. • Place all waste materials, including disposable PPE, into an autoclavable biohazard bag. Be careful not to contaminate the outside of the bag. REPORT ALL SPILLS TO YOUR SUPERVISOR AND THE BIOSAFETY OFFICER

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

26. 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.

27. Security • 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. Special security measures apply to users of Select Agents. See the Biosafety Officer for more specific details.

28. 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 4th Edition, May 1999 http://bmbl.od.nih.gov/ Guidelines for Research Involving Recombinant DNA Molecules (NIH Guidelines) http://www.niehs.nih.gov/odhsh/biosafe/nih/rdna-apr09.pdf

29. Resourcescontinued Material Safety Data Sheets For Research Pathogens http://www.hc-sc.gc.ca/hpb/lcdc/biosafty/msds/index.html Purdue University Biosafety Manual http://www.adpc.purdue.edu/PhysFac/rem/home/booklets/bioman.htm

30. Quiz Please take the following review quiz, then check your answers on the last slide. 1) Why is it important for lab staff to have bio- agent awareness training? a)  Some agents can cause serious complications for pregnancies and immuno-compromised individuals. b)  Signs and symptoms can determine if an exposure has taken place. c)  De-con procedures can be determined at that time. d)  All of the above.

31. 2)While working with an organism that has an exposure route including skin contact and respiratory, what personal protective equipment (PPE) is needed? a)  Biosafety cabinet b)  Respiratory or N-95 mask c)  Protective outer wear, gloves and goggles d)  All of the above 3) Why is posting a Hazard Assessment important ? a)  It is an accessible information source for employees to use to determine proper personal protective equipment. b)  It is required by the Center For Disease Control. c)  It is an ineffective bureaucratic hoop to jump.

32. 4) What is inappropriate laboratory personal protective equipment? a)  Gloves b)  Open toed shoes c)  Goggles 5) Sharps are defined as anything that will cut, puncture, or abrade the skin. True False 6) What should be done in the event of a bio-exposure sharps injury? a)  Wash the site with soap and water b)  Inform your supervisor c)  Contact REM d) All of the above

33. 7) Biohazard bags should not be…. a) overfilled. b) disposed of in the regular trash. c) held against your body or squeezed d) All of the above 8) If a spill occurs inside of a Biological Safety Cabinet, which of the following statements is not appropriate procedure to follow? a) Report the spill to your supervisor immediately. b) Clean it up fast, before anyone sees it. c) Wear the proper Personal Protective Equipment for the material for which you are working. d) Plan in advance for an emergency. For example, what supplies and equipment should you maintain in your area to assist you in the event of an accidental spill?

34. 9)Who is responsible for security in a BSL2 or higher level laboratory? a) Purdue Police b) The supervisor c) The Building Deputy d) Everyone is responsible for security in the laboratory. 10) Who should be contacted for questions about Biosafety related issues? a) University Police b) Biosafety Officer c) Building Deputy d) Radiation Safety Officer

35. QUIZ Answer Key 1) D 8) B 2) D 9) D 3) A 10) B 4) B 5) T 6) D 7) D