Biosafety in the Workplace

1. Biosafety in the Workplace PLS 4/595D /Regulations and Laboratory Management Spring Semester, 2006 Mark J. Grushka, M.S., CSP Manager, Biosafety and Biosecurity University of Arizona

2. Main Topics • Part 1 Introduction to Biological Safety Principles (Tuesday/April 11th) • Part 2 Introduction to Regulatory Framework (Tuesday/April 11th) • Part 3 Biosafety Program Management, Application of Project Management Techniques and Case Studies

3. Part 1 Introduction to Biological Safety Principles • Definitions • Key Principles • Data on Laboratory Acquired Infections • Risk Assessment • Biosafety Containment Levels • Primary Containment • Emergency Preparedness

4. Introduction to Biosafety Principles • Complex relationship between organisms and hosts. We are surrounded by countless microorganisms. Our bodies depend upon them for natural processes such as digestion. But most of time, we do not get sick because of natural defenses. • Infectious (pathogenic) organisms must: • Attach and survive hosts defenses • Multiply • Create signs and symptoms of disease in host

5. Definitions • Safety • Risk • Biosafety • Biosecurity

6. Safety • Freedom from harm • Control of accidental losses involving • People • Property • Loss to process

7. Risk • The chance that something may or may not happen. Often defined as: • Frequency (how often) • Severity (how bad)

8. Biosafety • “Development and implementation of administrative policies, work practices, facility design, and safety equipment to prevent transmission of biological agents to workers, other persons or the environment” MMWR December 6, 2002

9. Biosecurity • “Protection of high-consequence microbial agents and toxins, or critical relevant information, against theft, or diversion by those who intend to pursue intentional misuse” MMWR December 6, 2002

10. Koch’s Postulates • 1890 Robert Koch Established List of Criteria to Judge Whether or Not a Given Microbe Was Responsible for a Given Disease • The organism must be present in every case of the disease • The organism must be isolated from the diseased host and grown in pure culture • The specific disease must be reproduced when the pure culture is inoculated into a healthy susceptible host • The organism must be recovered from the experimentally infected host

11. Biohazardous Materials • Include All Infectious Organisms (Bacteria, Chlamydiae, Fungi, Parasites, Prions, Rickettsias, Viruses) which can cause disease in humans or cause significant environmental or agricultural impact. • Materials that may harbor infectious organisms such as human or primate tissues, fluids, cells, cell cultures.

12. Key Principles of Biosafety • Laboratory Practices and Techniques • Hand Washing Important • Manipulation of Material to Minimize Aerosols • Consistent Use of Personal Protective Equipment • Safety Equipment (Primary Barriers) • Biological Safety Cabinets (BSC’s) • Facility Design and Construction (Secondary Barriers/Room Design) • Room Pressure Negative to Corridor • Controlled Access to Non-Research Personnel • Medical Surveillance

13. Typical Routes of Entry for Viral or Bacterial Pathogens • Inhalation • Ingestion • Injection • Needle sticks • Accidental cuts with sharp objects • Skin or Eye Exposure

14. Laboratory Acquired Infections • Risk of Laboratory Associated Infections (LAI’s) is Real • Historical Examples Include:Brucellosis, Q Fever, Hepatitis, Typhoid Fever, Tuberculosis, Hepatitis • Of the 3921 LAI Only 703 (18%) Caused by Identifiable Accidents including needle sticks, broken glass, spills or sprays (R.M. Pike 1976)

15. Agent No. of Case No. of Deaths No. of Agents Involved No. of Published Cases Bacteria 1704 71 37 744 Viruses 1179 55 85 915 Rickettsiae 598 25 8 381 Fungi 354 5 9 313 Chlamydiae 128 10 3 71 Parasites 116 2 17 74 Totals 4079 168 159 2498 40 Years of Data on Overt LAI’sPike, R.M. 1978 Various Classes of Agents

16. Infection No. of Cases No. of Deaths Brucellosis 426 5 Q Fever 280 1 Hepatitis 268 3 Typhoid Fever 258 20 Tularemia 225 2 Tuberculosis 194 4 Dermatomycosis 162 0 Venezuelan Equine Encephalitis 146 1 Psittacosis 116 10 Coccidiodomycosis 93 2 Totals 2168 48 Ten Most Frequently Reported LAI’sPike, R.M. 1978 Past and Present Hazards of Agents

17. University’s Responsibilities • To provide a workplace free of recognized hazards. UA Policy and OSHA Requirement • To provide training to employees in order to recognize hazards and to protect employees against those hazards. • Methods of controlling risk may include: • Building Design Including Containment Features (Primary/Secondary) • Policies/Procedures (SOP’s) • Personnel Protective Equipment • Medical Surveillance Programs

18. Basic Risk Assessment Framework • Hazard Identification • Estimate Probability of Occurrence • Decide on Acceptable and Non-Acceptable Practices • Implement Practices • Monitor

19. Example of Risk Assessment for Cell Culture • BELGIAN BIOSAFETY SERVER http://www.biosafety.be/CU/animalcellcultures/mainpage.html – Introduction • Bioline International http://www.bioline.org.br/request?by95008

20. Risk Assessment Flow Chart

21. Employee Responsibilities • If you don’t know, ask. • If you have not been trained to do it, don’t! • Follow established biosafety practices and procedures. Always ask Principal Investigator. • Immediately inform Principle Investigator or Laboratory Manager if any accidents, spills, procedural issues/concerns or any questions arise about your safety or the safety of others.

22. Biosafety Levels Defined • BSL-1 • BSL-2 • BSL-3 • BSL-4

23. Biosafety Level One (BSL-1) • BSL-1 Work with Well Characterized Agents Not Known to Cause Disease in Healthy Adults. Standard Microbiological • Open bench tops acceptable with good standard microbiological practices • Laboratory not necessarily separated • Special containment equipment or facility design not required • Examples include E. coli K-12, Bacillus subtilis Also Called “Bench Work”.

24. Biosafety Level Two (BSL-2) • BSL-2 Work with Moderate Potential to Affect Personnel and Environment. (Herpes, Influenza viruses, Legionella sp.) • Personnel are specifically trained to handle pathogenic agents • Lab access limited when work is conducted • Extreme precautions taken when handling contaminated sharp items (needles, scalpels) • Appropriate immunizations are administered when available and baseline serum samples encouraged • Certain procedures require biological safety cabinets

25. Biosafety Level Three (BSL-3) • BSL-3 Work May Cause Serious or Potentially Lethal Disease as a Result of Exposure to Inhalation Route. (M. Tuberculosis, Bacillus anthraces) • Very specific training • Biosafety Cabinets used • Appropriate PPE and other clothing • Specific engineering and design features

26. Additional (BSL-3) Requirements • Immunization and medical surveillance protocols required • No open bench work • Ducted exhaust air ventilation creates directional airflow from “clean” toward “contaminated” areas prior to discharge to outside • High Efficiency Particulate Air (HEPA) filters may be required for room exhaust

27. Biosafety Level Four (BSL-4) • BSL-4 Work with dangerous and exotic agents which pose a high risk of aerosol-transmitted laboratory infectious and life threatening disease. Ebola, Marburg, • Special facility design features required • All activities confined to Class III biosafety cabinets (glove boxes), or Class II BSC’s used by workers using one piece positive pressure personnel suits ventilated by a life support system

28. Identifying Biohazard Risks • What am I Working With? How Can it Cause Disease and How do I Protect Myself? • Routes of Entry Include Inhalation, Ingestion, Inoculation, Skin and Eyes • Typical Risks of Exposure Include Contaminated Needles, Mouth-Pipetting, Splashing, Animal Bites

29. How to Protect Yourself • Knowledge and Understanding of the Biohazards You Are Working With • How Can it Get Onto/Into My Body • How to Protect Myself (Hierarchy of Control) • Containment Equipment • Techniques • Personal Protective Equipment

30. Identifying Biohazard Risk is Key • Accident/Incident Preceded Events Represented Only 18% of LAI’s • Aerosols, Droplets and Fomites are Likely Sources • Lab Techniques With High Potential for Exposure Include: • Centrifuges/Blenders, Opening Tubes/Bottles, Syringes/Needles, Inoculating Loops, Heating Over Flames

31. Mammalian Tissue Culture Work • Risks • Tissue culture may contain virus or bacteria capable of spreading to human host • Integrity of culture may be altered because of contamination from outside source • How to Reduce Risks to Human and Cell Culture? • Manipulation of tissue cultures only under Class II Biological Safety Cabinets • Use care when doing any procedure using instruments that may break skin • Use proper PPE like latex gloves, eye protection

32. Class II Biological Safety Cabinets Explained • Main Function • Protects Worker • Protects Work (Tissue Cultures From Microbial Contaminants, i.e.. Integrity of Cultures) • Features • High Efficiency Particulate Air (HEPA) Filter Minimizes Escape of Contaminants Within Cabinet Into Lab • HEPA Filtered Air Supply Bathes Work Surface, Protecting Work • Certified Annually by Facilities Management

33. Basis of Primary ContainmentIsolate the Laboratory Worker from Biological Agent With Ultra Filtered Directional Air Currents

34. Class II Type B1 Biological Safety Cabinet Air Flow/HEPA Filter Placement

35. Proper Use of Biosafety Cabinets • Do’s • Become familiar about the equipment by reading users manual and asking PI. • Keep laboratory doors closed and minimize movement in front of cabinet to avoid disrupting airflow. Avoid rapid arm movement in and out of BSC. • Decontaminate work surfaces with disinfectant before and after working in a cabinet according to laboratory standard operating procedures (SOP’s).

36. Proper Use of Biosafety Cabinets • Don’ts • Do not use cabinets as a permanent storage area for supplies (disrupts airflow) • Do not work inside cabinet with UV lamp on, if so equipped. (skin/eye burns) • Do not rapidly insert or withdraw arms. (disrupts airflow) • Place required equipment or supplies for procedure inside before beginning work. (minimizes hand/arm withdrawals which can disrupt airflow)

37. Eagleston Institute Biosafety Cabinet Clips

38. What Does Your Lab Look Like? Advantages of Good Housekeeping • Reduces Risk of Slip, Trip and Falls • Protects Integrity of Biological Experiments by Providing Adequate Space and Reduce Contamination Potential • Easier to Decontaminate Surfaces • Saves Time by Being Able to Find Stuff

39. Emergency Preparedness • What Should I Do When Things Go Wrong? • Learn the types of emergencies that could happen • Spills of liquids • Equipment malfunctions • Exposure to potential pathogens through inhalation, ingestion, skin including eye exposure, needle or other sharps • Learn how to respond to minimize exposure time and concentration • Contact your supervisor to protect your health and legal rights

40. Biohazard Spills • Each Lab Required to Have Spill Decontamination Plan • PI Required to Have Cleanup/Decon Procedure for Specific Biohazards Found in Lab • If Spill Occurs/General Guidelines • Remove affected clothing/gloves • Wash contaminated body areas with soap/H2O • Secure area until cleanup completed • Call UA Risk Management 621-1790 for technical assistance

41. Summary • Risks of Working with Biological Materials in Research are Real • The Risks Can be Managed Through: • Properly Identifying and Assessing Biological Risks • Good Laboratory Practice and Technique • Correct Use of Safety Equipment (Primary Barriers) Including Biological Safety Cabinets • Facility Design, Construction and Maintenance (Secondary Barriers) • Additional Resources are Available Through Institutional Biosafety Committee and Professional Staff

42. Regulatory Framework PLS 4/595D /Regulations and Laboratory Management Spring Semester, 2006 Mark J. Grushka, M.S., CSP Manager, Biosafety and Biosecurity University of Arizona

43. Part 2 Introduction to Introduction to Regulatory Framework • What are the major regulations covering biosafety? • How is the University of Arizona organized for biosafety compliance? • What are the future implications for regulatory control of biosafety?

44. Introduction • The regulatory framework covering biosafety can be characterized as a combination of statutes, regulations, rules and guidelines from various federal and state agencies, private and public organizations and other interested parties such as manufacturers of containment equipment

45. Federal Laws Occupational Health and Safety Act (OSHAct) • Bloodborne Pathogens(29 CFR 1910.1030) • Occupational Exposure to Hazardous Chemicals in Laboratories(29 CFR 1910.1450) • Personal Protective Equipment(29 CFR 1910.132-139) • Needlestick Standard

46. National Institutes of Health • NIH Office of Biotechnology Activities • NIH Guidelines for Research Involving Recombinant DNA Molecules • IBC Resources

47. USDA • APHIS

48. USDOT • HazMat Safety • Hazardous Materials Regulations (49 CFR 100-185)

49. US EPA • Hazardous Waste • Microbiology • IAQ

50. Select Agents • All individuals who have access to Select Agents must undergo a Security Risk Assessment • Acquisition, use, transfer and disposal of Select Agents is monitored by CDC/APHIS through issuance of registration