BIOSAFETY INSPECTIONS. What to ask and what to look for

Transcription

1 BIOSAFETY INSPECTIONS What to ask and what to look for 1

2 Biosafety is risk-based, not necessarily rule-based however, there are a few rules and guidelines: NIH Guidelines for Research Involving Recombinant or synthetic Nucleic Acid Molecules (The Guidelines) Only a few actual rules!!! But we are bound to them because we accept federal $ Biosafety in Microbiological and Biomedical Laboratories (BMBL) - Not actual rules (but the standard ) CWA 15793: Laboratory Biorisk Management voluntary (soon to be an ISO) CWA=CEN Workshop Agreement CEN=European Committee for Standardization Dual Use Research of Concern (DURC) actual rules! Gain of Function Recommendations (GOF) - recommendations Federal Select Agent Program (FSAP) actual rules! OSHA BBP rules actual rules! 2

3 Where s the risk? Is the protocol registered with the Institutional Biosafety Committee (per the Guidelines)? Risk to the University & future funding The organism is it pathogenic? Personal/environmental/ecological risk? Modes of transmission mucous membrane? Aerosol transmissible? Recombinant nucleic acids are the genes oncogenic? antibiotic resistant? Is the vector Contagious? Can it replicate? Can it complement latent viruses? What tissues does it target? The procedure - Does it make sense? Engineering controls? The lab clean? Well maintained equipment? Proper procedures for used items, RMW 3

4 With Risk in mind, when doing an inspection what should you look for? UB checklist for BSL-2 labs /1/f_ Based on the DHHS s Biosafety in Microbiological and Biomedical Laboratories guidelines (which is the Biosafety Bible). 4

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6 Inspection Topics Standard Laboratory Practices Special Practices Laboratory Equipment Laboratory Facility 6

7 Standard Practices Access controlled by a lab supervisor Wash hands after working with agent and before leaving the lab Eating, drinking, smoking, applying makeup or contact lens is prohibited Mechanical pipettes are used Sharps policy (prescriptive) in place Minimize splashes Decontaminate work surfaces Decontaminate all material before disposal Lab signage posted at entrance Lab supervisor ensures adequate training of all personnel 7

8 Special Practices Lab personnel must be provided appropriate medical surveillance & offered immunizations if available Anyone entering the lab is to be notified of the hazards within the lab A biosafety manual must be prepared, adopted and in a visible/accessible location Supervisor must ensure proficiency in standard and special microbiological practices Potentially infectious materials must be placed in a durable, leak proof container during collection, handling, processing, storage, or transport within a facility Lab Equipment should be routinely decontaminated and must be decontaminated after spills, leaks or other exposures 8

9 Special Practices cont. Spills must be contained, decontaminated and cleaned by appropriately trained and equipped staff Equipment must be decontaminated prior to maintenance or removal Exposures must be reported to EHS and treated appropriately by medical staff Animals and plants not associated with research are not permitted Aerosols must be contained in a BSC or by other physical means 9

10 Safety Equipment BSCs, PPE and other containment devices must be used when: Procedures with a potential for creating infectious aerosols or splashes are conducted. These may include pipetting, centrifuging, grinding, blending, shaking, mixing, sonicating, opening containers of infectious materials, inoculating animals intranasally, and harvesting infected tissues from animals or eggs. High concentrations or large volumes of infectious agents are used. Such materials may be centrifuged in the open laboratory using sealed rotor heads or centrifuge safety cups Protective laboratory coats, gowns, smocks, or uniforms designated for laboratory use must be worn while working with hazardous materials. Remove protective clothing before leaving for non-laboratory areas. Dispose of protective clothing appropriately, or deposit it for laundering by the institution 10

11 Safety Equipment cont. Eye and face protection (goggles, mask, face shield or other splatter guard) is used for anticipated splashes or sprays of infectious or other hazardous materials when the microorganisms must be handled outside the BSC or containment device. Gloves must be worn to protect hands from exposure to hazardous materials. Change gloves when contaminated, glove integrity is compromised, or when otherwise necessary. Remove gloves and wash hands when work with hazardous materials has been completed and before leaving the laboratory. Do not wash or reuse disposable gloves. Dispose of used gloves with other contaminated laboratory waste. Eye, face and respiratory protection should be used in rooms containing infected animals 11

12 Laboratory Facility Laboratory doors should be self-closing and have locks Laboratories must have a sink for hand washing The laboratory should be designed so that it can be easily cleaned and decontaminated. Laboratory furniture must be capable of supporting anticipated loads and uses. Laboratory windows that open to the exterior are not recommended. However, if a laboratory does have windows that open to the exterior, they must be fitted with screens BSCs must be installed so that fluctuations of the room air supply and exhaust do not interfere with proper operations. BSCs should be located away from doors, windows that can be opened, heavily traveled laboratory areas, and other possible airflow disruptions 12

13 Laboratory Facility cont. Vacuum lines should be protected with liquid disinfectant traps new facilities should consider mechanical ventilation systems that provide an inward flow of air without recirculation to spaces outside of the laboratory HEPA filtered exhaust air from a Class II BSC can be safely recirculation back into the laboratory environment if the cabinet is tested and certified at least annually and operated according to manufacturer s recommendations A method for decontaminating all laboratory wastes should be available in the facility (e.g., autoclave, chemical disinfection, incineration, or other validated decontamination method) 13

14 That s a lot, how do we get there? Ask questions PI Students Lab Manager Custodian Inspect Lab benches Environmental chambers Look at plates and culture vessels 14

15 What to ask? What is your research focused on? What organisms are you using? Is it a virulent strain? What is your gene of interest? What kits or other procedures are you using? Is the protocol registered with the IBC? 15

16 What is the focus or your research? Listen for names of bacteria or viruses Listen for the species Humans? Animals? Can anything escape the lab? antibiotic resistance genes? arthropods? Listen for the genes of interest Listen for vectors (plasmids, AAV, CRISPR/Cas) 16

17 Organisms Bacteria, viruses, human or animal cell lines? Bacteria are on agar plates (top picture) or broth culture Cell lines and viruses are grown in cell culture (bottom left) 17

18 Is it a virulent strain? All organisms come in different flavors, some strains of E. coli will make you sick, others are beneficial and already live inside you Typically, virulence factors reside on mobile genetic elements such as plasmids or bacteriophage 18

19 Humans or Animals Are they approved by the IRB (Humans, even blood) or the IACUC (all vertebrate animals)? 19

20 Kits, Chemicals & Equipment Each kit comes with its own dangers, depending on the chemicals and the procedure Equipment fails, problems arise. 20

21 Is your work registered with the IBC? All work with pathogens and recombinant nucleic acids must be registered with the IBC (exempt work included) at UB Not all Universities have the same rules! 21

22 What else? Plants or animals not associated with research? Proficient work practices? PPE (on people, in the room)? Engineering controls? Is there a sink near by? Food or drinks? Disinfectant near by? Equipment cleaned out? 22

23 Biosafety Cabinet Certification All BSCs in use should be certified annually Certification stickers are specific to each cabinet Info includes the type of cabinet, its serial #, date of inspection, next inspection date and the certifier UB EH&S has the contact info for the companies that perform BSC certifications in the area 23

24 What else? Vacuum Traps need to be attended to, do not let things grow in them and place them in secondary containment when used in a BSC. 24

25 What else? Where do used tips go? Any sharps (needles, razors) laying 25

26 What is regular trash and what is regulated medical waste??? 26

27 Caption: digniss and in aliquet ni et umis varius. 27

28 Sharps Containers Sharps containers should be rigid, puncture-resistant, and leak proof. They also need to be sealable and have the correct markings. They should not be found in hallways or stairwells. They should be in labs under the control of the lab manager. They should not be made out of popbottles or other odd containers 28

29 Wow, that s a lot, what can be ignored? Closed plates on a bench or BSC Many reagents without GHS labelling (PBS, Coomassie stain, TBE, etc.) Secured RMW Sometimes, its hard to know where to start and when to finish BE REASONABLE Upon close inspection, nothing in this mess was inherently dangerous with the exception of the mess itself. 29

30 Example 1 IGNORE the stained bench paper. Its probably just a buffer solution and it is dry. 30

31 Example 2 Do NOT ignore balancing RMW containers on bench tops, they do not belong there. 31

32 Example 3 Ignore plates that have nothing growing on them. Laugh at the student who piled them this high when they fall and he/she needs to make new ones. 32

33 Example 4 Temporary RMW containment The full bag of RMW is fine where it is, it will be thrown into a larger box later. The 95% ethanol bottle does not have GHS labeling 33

34 Example 5 Don t ignore used gloves just have the student throw them out The plate of bacteria is fine so long as its not a highly virulent strain of something. 34

35 Example 6 Ugh! Clearly, these folks are wearing contaminated gloves when working at this site. We decided to make a rule to always wear gloves when working here but, it still had to be cleaned up. 35

36 Example 7 Animal organs They are labelled and in a storage medium, ignore. 36

37 Example 8 Animal feces and urine In a CO2 asphyxiation chamber, do not ignore, make them clean it out, IACUC would. 37

38 Example 9 Enough said Do not ignore, this is not really consistent with any thought for safety (or even good science). 38

39 Kids will be kids Watch out for playful activities They may be fun and test your skillz but clearly do not fit with safe science Dry Ice bombs and rockets are always a favorite 39

40 When in doubt, check with the BSO! A Special Thank you to Dr. Nicole Gerber for pitch hitting! Dr. David Pawlowski drp@buffalo.edu 40