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East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Biological Safety.

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Presentation on theme: "East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Biological Safety."— Presentation transcript:

1 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Biological Safety Training

2 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Introduction to Biological Safety Biological Hazards are divided into 4 Biosafety Levels –BSL 1 –BSL 2 –BSL 3 –BSL4 Biosafety levels define the lab requirements, protective clothing, and work practices

3 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety BioSafety Definitions BSL 1 (BMBL) –Not known to consequently cause disease in healthy human adults –Pose minimal hazards under ordinary conditions of handling

4 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety BioSafety Level 1 Standard Practices –Use Mechanical Pipetting devices –No Eating, Drinking, Smoking in Lab –Minimize splashes and aerosols –Decontaminate work surfaces –Safe handling of sharps –Wash Hands before leaving lab

5 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety BioSafety Level 1 Protective Clothing –Lab Coat –Gloves –Eye Protection

6 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety BioSafety Definitions BSL 2 (BMBL) –suitable for work involving well-characterized agents not known to cause disease in healthy adult humans, and of minimal potential hazard to laboratory personnel and the environment Examples: Measles Virus, Salmonella, Hepatitis B Virus

7 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety BioSafety Definitions Other Materials Handled at BSL 2 (Requires compliance with OSHA BBP Standard) –Human Blood –Human Blood Components –Human Tissues –Human Cell Lines

8 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety BioSafety Level 2 Standard Practices –All requirements for BSL 1 plus: Access to laboratory is limited or restricted when work is being conducted Personnel have specific training in handling pathogenic agents Biohazard Sign posted on the door Extreme precautions are taken with contaminated sharp items

9 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety BioSafety Level 2 Standard Practices cont. Written Biosafety Procedures Report Spills Minimize aerosol generation Personnel receive appropriate immunizations or test (e.g. Hep B vaccine or TB skin Testing)

10 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety BioSafety Level 2 Written Biosafety Procedures –Prepared specifically for lab by PI –Incorporated into Lab Safety Plan –Personnel are advised of special hazards –Personnel are required to read and follow procedures

11 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety BioSafety Level 2 Personal Protective Equipment –Lab Coats –Gloves Double Gloving when appropriate –Eye Protection - Safety Glasses/Shield

12 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety BioSafety Level 3 BSL 3 (BMBL) –applicable to clinical, diagnostic, teaching, research, or production facilities in which work is done with indigenous or exotic agents which may cause serious or potentially lethal disease as a result of exposure by the inhalation route.

13 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety BioSafety Level 3 Standard Practices –All requirements for BSL 2 plus: Limited lab access 2 doors in a series to access lab Able to wash entire lab Special exhaust ventilation (Not re circulated, negative pressure)

14 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety BioSafety Level 3 Standard Practices –cont. –High level of training –Personnel receive vaccinations if available –Work in Biosafety cabinets

15 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety BioSafety Level 3 Examples –Mycobacterium Tuberculosis –Hanta virus –SARS

16 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety BioSafety Level 4 BSL 4 (BMBL) –required for work with dangerous and exotic agents which pose a high individual risk of aerosol-transmitted laboratory infections and life-threatening disease.

17 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety BioSafety Level 4 Requirements –All requirements for BSL3 plus: Class III Biosafety cabinet or positive pressure suits Shower/Change rooms Clothing Autoclaved before laundering Air Locks

18 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety BioSafety Level 4 Examples –Ebola Virus –Monkey B Virus –Marburg Virus

19 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Containment Primary –Protection of personnel and immediate lab from exposure Good Techniques Safety equipment –Biological Safety Cabinet –Engineering Controls –PPE –Serologic surveillance

20 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Containment Secondary –Protection of environment external to lab Facility Design Operational practices

21 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Safe Work Practices for all Levels Wash hands after work; when removing gloves; before leaving lab No eating, drinking, applying cosmetics, handling contact lenses in the lab. Maintain labs in clean, orderly fashion.

22 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Safe Work Practices for all Levels Limit access to lab when work with organisms is in progress Use good microbiological techniques No mouth pipetting When possible use plastic instead of glass

23 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Sharps Safety Sharps include needles, syringes, razor blades, lancets, slides, scalpels, pipettes, micropipettes, pipette tips, broken plastic or glassware, and other devices capable of cutting or piercing the skin.

24 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Sharps Safety Contaminated needles shall not be bent, recapped, or removed unless there is no feasible alternative. If required, use a mechanical device or a one handed technique.

25 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Sharps Safety Safety devices or alternatives to needles should be used when available.

26 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Sharps Disposal Sharps containers for disposal of these items should be conveniently located and easily accessible in all work places where sharps are used.

27 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Sharps Disposal Syringes and syringes without a needle attached go into a sharps container Contaminated micropipettes, pipette tips, and Pasteur pipettes are discarded in a puncture-resistant container or a sharps container for disposal.

28 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Sharps Disposal Dont place needles or sharps in office waste containers

29 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Laminar Flow Equipment BioSafety Cabinets (BSCs) contain infectious agents to protect personnel and the environment Laminar Flow Clean Benches (LFBs) Non Hazardous work only Protect work from contamination

30 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety BioSafety Cabinets 3 Classes All exhaust is HEPA filtered before leaving the cabinet Class I –Do not protect the work from contamination –Air entering cabinet is not filtered

31 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Biosafety Cabinets Class II (4 types – A, B1, B2, and B3) –Each type recirculates different amount of air –Some are hard ducted, and some exhaust into the room. Class III Totally enclosed, ventilated cabinets Work through portals with attached gloves

32 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety BioSafety Cabinets All equipment is laid out to not restrict airflow in the cabinet

33 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Laminar Flow Benches Do not protect personnel or the environment Discharges HEPA filtered air across the work surface toward the user.

34 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Use of Laminar Flow Equipment Minimize airflow disturbances (moving in/out of cabinet, people walking by, opening doors, blocking grills with equipment) Wipe down surfaces with approved disinfectant before and after work.

35 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Use of Laminar Flow Equipment Minimize use of flames in cabinet Try not to use chemicals in Class I or II hoods that are ducted Do not store excess equipment in cabinet All laminar flow equipment must be certified annually.

36 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Centrifuges

37 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Centrifuge Hazards Mechanical failure of Machine Lab equipment failure (tubes etc.) Aerosol generation Operator error

38 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Centrifuge Operation 1.Check tubes for cracks/chips 2.Use matched sets of tubes, buckets, etc. Make sure the rotor is balanced properly 3.Tightly seal all tubes and safety cups 4.Ensure that rotor is locked to spindle and the bucket is seated 5.Close lid during operation 6.Allow to come to complete stop before opening

39 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Centrifuge Safety Use sealed tubes, safety buckets or rotors when possible When possible fill and open centrifuge tubes or buckets in the biological safety cabinet. Lubricate O-rings and rotor threads weekly Disinfect weekly and after all spills or breaks

40 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Biohazardous Waste –All cultures, stocks, and other regulated waste are decontaminated before disposal by an approved decontamination method such as autoclaving.

41 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Biohazardous Waste Examples –stock cultures –materials containing or contaminated with blood –blood and body fluids –sharps –pipette tips –animal waste –other contaminated lab material

42 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Blood and Body Fluids Waste Serum, Plasma Other blood components Semen Vaginal secretions Cerebrospinal fluid Pleural fluid Peritoneal fluid Pericardial fluid Amniotic fluid Any other body fluid visibly contaminated with blood

43 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Infectious Sharps Waste Hypodermic needles Scalpels Pipettes /pipette tips Breakable containers Glass products (i.e., slides or cover skips)

44 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Biohazardous Waste Inappropriate materials Dispose of noninfectious waste in regular trash

45 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Biohazardous Waste Inappropriate materials These include: aerosol cans, chemical waste, flammable material

46 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Biohazardous Waste Inappropriate packaging Use puncture resistant package if needed

47 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety

48 East Carolina University Brody School of Medicine Office of Prospective Health/Biological Safety Biohazardous Waste Containers Biohazardous waste containers shall be clearly marked with the universal biohazard symbol.


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