BMBL and BBP Overview

1. BMBL and BBP Overview Environmental Health & Safety 713-500-8100

2. Why Biological Safety • “An ounce of prevention is worth a pound of cure.” Benjamin Franklin • Harding & Byers (2000) • 1,267 overt LAIs • 22 deaths (5 were aborted fetuses) • 663 additional subclinical infections

3. Overview • Definition & history of biological safety • Regulations & guidelines • Principles of biosafety • Risk assessment (& biological agents) • Containment • Biological safety levels (BSL-1 thru BSL-4) • Bloodborne pathogens (BBP)

4. Biological Safety • The field of biosafety promotes safe laboratory practices, procedures, along with proper use of containment equipment and facilities; also provides advice on laboratory design.

5. Biological Safety • Biosafety - the discipline addressing the safe handling & containment of infectious microorganisms & hazardous biological materials • application of knowledge & the use of appropriate techniques & equipment • prevent personal, laboratory & environmental exposure to potentially infectious agents or biohazards Source: BMBL, 5th Ed.

6. Biological Safety - History • Laboratory-associated infections (LAIs) first reported ~ start of 1900s • Pike & Sulkin (1951) studies identified • 4,079 LAIs reported (1930 – 1978) • 168 deaths • 10 most common causative agents Source: BMBL, 5th Ed.

7. Biological Safety - History • Next 20 yrs • Harding & Byers (2000) worldwide literature search • 1,267 overt LAIs • 22 deaths (5 were aborted fetuses) • Most common causative agents • 663 additional subclinical infections Source: BMBL, 5th Ed.

8. Biological Safety - History • Small number of specific incidents identified • Non-specific: • working with a microbiological agent • being in or around the lab • being around infected animals • Possible contributing factors to reduced LAIs: • improvements in containment equipment, engineering controls, & greater emphasis on safety training Source: BMBL, 5th Ed.

9. Biological Safety - History • US Army Biological Research Laboratories, Fort Detrick (pioneering in biosafety, 1944 - 1969) • Biosafety programs at USDA, National Animal Research Center, DHHS, CDC, NIH • 1974 – CDC & NIH publications (agent classification, research safety) • 1976 – NIH Guidelines 1st published • 1984 – biosafety principles introduced in BMBL, 1st Ed. Source: BMBL, 5th Ed.

10. Regulations & Guidelines • Biosafety in Microbiological and Biomedical Laboratories, 5th Edition, CDC/NIH (BMBL) • NIH Guidelines for Research Involving Recombinant DNA Molecules, April 2002 as amended • OSHA Bloodborne Pathogens Standard 29 CFR 1910.1030 • Respiratory Protection including Tuberculosis 29 CFR 1910.134 (independent standard 1910.139 repealed) • State regulations concerning biological waste disposal • IATA Dangerous Goods Regulations, DOT 49 CFR 171-185, & USPS 39 CFR Part 111 • Select Agents and Toxins Regulations (HHS/CDC 42 CFR 73; USDA/APHIS 7 CFR 331 & 9 CFR 121)

11. Areas of Biosafety • Laboratory Safety • Bloodborne pathogens (BBP) & other infectious agents • Recombinant DNA (rDNA) • Biological waste disposal • Infectious substance shipping • Respiratory Protection (TB) • Bioterrorism & select agents/toxins • Mold & indoor air quality • Occupational safety & health in the use of research animals

12. Biosafety in Microbiological and Biomedical Laboratories (BMBL), 5th Edition (2007) • Biological Risk Assessment (pg. 9) • Principles of Biosafety (pg. 22) • Laboratory Biosafety Level Criteria (pg.30) • Vertebrate Animal Biosafety Level Criteria (pg. 60) • Principles of Biosecurity (pg. 104) • Occupational Health & Immunoprophylaxis (pg. 114) • Biological Agent Summary Statements(pg. 123)

13. Principles of Biosafety • Containment • microbiological practices, safety equipment & facility safeguards that protect laboratory workers, the environment & the public from exposure to infectious agents that are handled & stored in the laboratory • Risk assessment • process that enables the appropriate selection of microbiological practices, safety equipment & facility safeguards that can prevent laboratory-associated infection (LAI)

14. Biological Risk Assessment • Process used to identify: • Hazardous characteristics of a known infectious or potentially infectious agent or material (agent hazards) • Activities that can result in a person’s exposure to an agent (lab procedure hazards & capability to control hazards) • Likelihood that such exposure will cause a laboratory-associated infection (LAI) • Probable consequences of such an infection

15. Biological Risk Assessment • Principal hazardous characteristics of an agent: • Capability to infect & cause disease in a susceptible human or animal host • Virulence as measured by severity of disease • Availability of preventive measures & effective treatments for the disease

16. Biological Risk Assessment • Other hazardous characteristics of an agent: • Probable routes of transmission of laboratory infection • Infective dose • Stability in the environment • Host range • Endemic nature

17. Hazard Classifications of Microbial Agents • Risk Group 1: Agents not associated with disease in healthy adult humans (no or low individual risk). • Risk Group 2: Agents associated with human disease which is rarely serious & for which preventive or therapeutic interventions are often available (moderate individual risk). • Risk Group 3: Agents associated with serious or lethal human disease for which preventive or therapeutic interventions may be available (high individual risk). • Risk Group 4: Agents likely to cause serious or lethal human disease for which preventive or therapeutic interventions are not usually available (high individual & community risk).

18. Biological Agents by Size prions (smallest) viruses (0.05 - 0.1 micrometer) 0.3 micrometer = HEPA filter testing particle size bacteria (0.5 - 1.5 micrometers) red blood cell (5 micrometers) protozoa (1 - 300 micrometers) fungi (largest)

19. Prions • Proteinaceous infectious particles • Lack nucleic acid • Abnormal isoform of a cellular protein • PrPSc(protease-resistant protein, scrapieisoform) • Examples – Kuru, Creutzfeldt-Jakob disease (CJD), Scrapies, Chronic Wasting Disease, Bovine Spongiform Encephalopathy (BSE) • Crosses blood-brain barrier

20. Prions

21. Viruses • Structure - envelope, capsid, core, genome • Contain fundamental information (DNA or RNA) • Must utilize living cells machinery & enzymes to replicate or make copies of themselves

22. Bacteria • No membrane-bound nucleus or organelles • Circular DNA called plasmids (transmit traits such as antibiotic resistance) • Metabolically very diverse • Ecological importance: nutrient cycling, decomposition • Shapes - cocci, bacilli, spirilla • Cell wall - peptidoglycan • Flagella - tail for movement • Fimbriae - adherence • Pili – transfer of bacterial DNA

23. Fungi • Contain chitinous cell wall • Most grow as multicellular filaments called hyphae forming a mycelium, some grow as single cells • Obtain nutrients in an already synthesized form • They must live in or on their food • Some parasitic fungi can absorb nutrients directly from host • Other parasitic fungi & all saprophytic fungi must secrete enzymes to digest surrounding material before absorbing it

24. Biological Containment • Elements of Containment • Laboratory Practice & Technique • Safety Equipment (Primary Barriers & Personal Protective Equipment) • Facility Design & Construction (Secondary Barriers)

25. Biological Containment • Laboratory Practice & Technique • Strict adherence to standard microbiological practices & techniques • Personnel must be aware of potential hazards & must be trained • Biosafety or laboratory Standard Operating Procedures (SOPs)

26. Biological Containment • Safety Equipment (Primary Barriers) • Biological safety cabinets (BSC) • Safety centrifuge cups/sealed rotor heads • Personal protective equipment (PPE) • lab coats, gowns, gloves, safety glasses, face shields, shoe covers, respirators, Tyvek suit

27. Biological Containment Biological Safety Cabinets (BSC) Class I, Class II & Class III

28. Biological Containment • Facility Design & Construction (Secondary Barriers) • Separation of lab from public access • Controlled access zones • Separate buildings or modules • Decontamination facility (autoclave) • Specialized ventilation systems (directional airflow) • Air treatment systems to remove agents from exhaust air (HEPA filter units) • Airlocks as lab entrances

29. Biological Safety Levels (BSL) • Biosafety levels are combinations of lab practices, safety equipment and lab facilities • BSL-1 through BSL-4 • Ascending degree of protection provided to personnel, the environment & the community • Increasing levels of containment

30. Biological Safety Level - 1 (BSL-1) • Standard microbiological practices: • Hand washing facility present • No eating, drinking, smoking, applying cosmetics, handling contact lenses, or storing food • No mouth pipetting • Safe handling of sharps policy implemented • Decontaminate work surfaces after completion of work, & any spill or splash; any waste generated • Biohazard symbol when infectious agents present • Wear lab coats or gowns, protective eye wear, gloves • Windows to exterior have screens

31. Biological Safety Level – 2 (BSL-2) • All of BSL-1 practices plus: • Access is restricted to personnel trained to handle pathogenic agents when work is being conducted • Lab doors should be self-closing & have locks • BSC used when potential for aerosol production • Sealed centrifuge cups or sealed rotors are used • Eyewash station must be readily available • Method for decontaminating laboratory waste should be available in the facility

32. Biological Safety Level – 3 (BSL-3) • All of BSL-2 practices plus: • Lab is separated from unrestricted traffic flow • Access to lab is restricted to entry through a series of 2 self-closing doors (ante-room) • All manipulations must be conducted in a BSC, other containment, or when wearing proper PPE • Protective clothing with a solid front (tie-back or wrap-around gowns, scrubs, coveralls) is worn • Eye, face, & respiratory protection must be worn in rooms containing infected animals • Hands-free sink near exit door

33. Biological Safety Level - 3 (cont.) • Lab must be designed for easy cleaning & decon; floors, walls & ceiling should be sealed; floors should be slip resistant; windows must be sealed • Vacuum lines must be protected with HEPA filters or their equivalent, filters must be replaced as needed • Method for decontaminating laboratory waste should be available in the facility, preferably within the lab • Sustained directional airflow (“clean” to “potentially contaminated”), visual monitoring device at entrance • Lab exhaust air must not re-circulate to other areas of the building

34. Biological Safety Level – 3 (BSL-3) Lab

35. Biological Safety Level – 4 (BSL-4) • All of BSL-3 practices plus: • Generally a separate facility; dedicated mechanical, electrical, plumbing, HVAC • Agents handled in Class III BSC or with one-piece positive pressure personnel suit • Personnel enter via locked doors & clothing change room; & exit via sequential inner (dirty) change room, personal shower & outer (clean) change room • Lab clothing decontaminated prior to laundering • Liquid effluent decontaminated prior to release to sewer • Redundant supply fans recommended, redundant exhaust fans required, interlocked supply & exhaust

36. Appendices • Appendix A – Primary Containment for Biohazards (BSC) • Appendix B – Decontamination and Disinfection • Appendix C – Transportation of Infectious Substances • Appendix D – Agriculture Pathogen Biosafety • Appendix E – Arthropod Containment Guidelines • Appendix F – Select Agents and Toxins • Appendix G – Integrated Pest Management • Appendix I – Toxins of Biological Origin • Appendix J – NIH Oversight of Research Involving Recombinant Biosafety Issues • Appendix K – Resources for Information • Appendix L - Acronyms

37. Bloodborne Pathogens • Hepatitis B virus (HBV) infection is the major infectious bloodborne occupational hazard to healthcare workers. • It’s estimated that 8,700 infections occur each year in HCW according to The Hepatitis Branch of the Centers for Disease Control (CDC) • For HCW these HBV infections cause: • over 2,100 cases of clinical acute hepatitis • 400-440 hospitalizations • approximately 200 deaths each year in healthcare workers.

38. Bloodborne Pathogens • Death may result from both acute and chronic hepatitis. • One milliliter of HBsAg positive blood may contain 100 million infectious doses of virus; thus, exposure to extremely small inocula of HBV-positive blood may transmit infection. • In different studies, 7% to 30% of susceptible healthcare workers sustaining needlestick puncture injuries from HBsAg positive patients became infected if they did not receive post-exposure prophylaxis

39. Bloodborne Pathogens (BBP) • OSHA Bloodborne Pathogen Standard 29 CFR 1910.1030 http://www.osha.gov/SLTC/bloodbornepathogens/index.html • Originally issued in 1991 • In 2001 the Standard was revised with the Needlestick Reduction Act which includes: • Education and selection of sharps injury reduction devices (e.g., self-sheathing needles) • Maintenance of a contaminated sharps injury log

40. Bloodborne Pathogens (BBP) • Texas Department of State Health Services Bloodborne Pathogen Control • Prevention of contaminated sharps injuries, needlesticks • Exposure control plan designed to minimize exposure of governmental entity employees to bloodborne pathogens http://www.dshs.state.tx.us/idcu/health/bloodborne_pathogens/pathogen_control/

41. Appendix H of BMBL • Guidelines for Work with Human, NHP, and Other Mammalian Cells and Tissues • “Recommended Practices. Human and other primate cells should be handled using Biosafety Level 2 practices and containment.”

42. Bloodborne Pathogens (BBP) • Definition: • Pathogenic microorganisms that are present in human blood or other potentially infectious material (OPIM), and can infect and cause disease in humans. These microorganisms can be transmitted through contact with contaminated blood, body fluids or OPIM.

43. Bloodborne Pathogens (BBP) • Examples: • HIV • HBV • HCV • T. pallidum • Herpes Virus • M. tuberculosis (typically an aerosol hazard) • Human T-Lymphotropic Virus Type I (HTLV-I)

44. Bloodborne Pathogens (BBP) • Body fluids that can harbor BBP: • Blood • Semen & vaginal secretions • Saliva involved in dental procedures • Synovial fluid • Cerebrospinal fluid • Human tissue & cell cultures • All body fluids containing blood

45. Bloodborne Pathogens (BBP) Occupational Exposure* • Mucous membrane contact • Splash to the eyes, nose or mouth • Percutaneous inoculation • Misuse of sharps (broken glass, needles, scalpels) • Exposure to broken / damaged skin • Risk increases if contact involves a large area of broken / damaged skin or if contact is prolonged * Risk increases with high titer levels in the source

46. Preventive Measures • Risk of exposure can be minimized or eliminated by using the following: • Engineering controls • Personal protective equipment (PPE) • Administrative controls • Work place practices

47. Engineering Controls • Leakproof containers • Use for storage & transport ofALLBBP material • Sharps containers • Fill no greater than ¾ full • Needleless devices • Use retractable syringes, self-sheathing needles • Biosafety cabinet (BSC) • Directional air flow • High efficiency particulate air (HEPA) filtration • Access control

48. Personal ProtectiveEquipment • Face protection • Goggles or safety glasses with side shields • Clothing • Lab coats, scrubs, disposable gowns (long pants only and no open toed shoes) • Replaced immediately when contaminated • Lab clothing restricted to work area • Gloves • Replaced immediately if torn • Not worn outside the lab area!

49. Administrative Controls • Medical surveillance • TB skin test (PPD), Quantiferon test • Immunizations • Hepatitis B series • Training • Management of staff (SOP compliance) • Background checks, security clearance

50. Good Work Place Practices • Practice Universal / Standard Precautions • Treat all human blood & body fluids as if known to be infectious for HIV, HBV or other potentially infectious material (OPIM) ! !