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BSL3 To Infect

BSL3 To Infect. Inhalation Laboratory. System Animals. Team Members. Nick Anders – Team Leader Stacey Hoebel – Communications Jeff Swift – BWIG Maja Middleton – BSAC Kayla Ericson – BWIG Austin Ramme - BWIG. Advisor Professor Willis Tompkins Client Adel Talaat Assistant Professor

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BSL3 To Infect

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  1. BSL3 To Infect

  2. Inhalation Laboratory

  3. System Animals

  4. Team Members Nick Anders – Team Leader Stacey Hoebel – Communications Jeff Swift – BWIG Maja Middleton – BSAC Kayla Ericson – BWIG Austin Ramme - BWIG

  5. Advisor Professor Willis Tompkins Client Adel Talaat Assistant Professor Department of Animal and Biomedical Sciences

  6. Abstract • Our objective is to build a safe system for infecting small animals with a Biological Safety Level 3 pathogen. The design must incorporate safety features to prevent the escape of the infectious materials from the infection chamber to the surrounding environment. The research will be used to study pathogenesis of tuberculosis. Ultimate outcome of this study is the development of novel drugs and vaccines that can cure tuberculosis patients.

  7. Problem Statement Develop a safe inhalation system for animal infection Should be able to infect animals ranging in size from mice, rats, and guinea pigs to rabbits Should be equipped with pathogen escape prevention measures to insure safety in the surrounding environment

  8. Background • Client’s goals of animal infection study • To study the pathenogenesis of tuberculosis (BSL3) • To develop a cure for TB patients • Safety Precautions • Proper training and protective clothing • Non-porous material • Autoclave in work zone • HEPA filtration • Sealed exhaust system with directional airflow

  9. Current Device in Use • Main chamber: 75 cm x 50 cm x 15 cm • Inner chambers: radius 8 cm, height 12 cm • Nebulizer, Ethanol Trap, HEPA Filter • Tupperware

  10. Current Models available • Glas-Col Manufactures a model for $15,000 • Independent system • UV light and HEPA filters incorporated • Automated controls • UW-Madison Engineering Shops make a model for $33,000 • Independent system • Stainless Steel Cylinder • Automated Controls • Clamps for seal

  11. Design Specifications • Dimensions: • Length: 25 cm – 75 cm • Width: 40 cm – 30 cm • Height: 10 cm – 70 cm • Withstand the Autoclaving Process • Storage at Room Temperature • Air Tight Structure • Deliver 5 mL of culture in 20 minutes • Service Life: 50 to 100 Uses

  12. Air input Alternative Design 1: • Pros • Dual Chamber System • Split Air Flow • Latex Flange around Neck • Cons • Specific only to mice • Too difficult to manufacture • Too expensive animal Air output Animal head Air input

  13. Alternative Design 2: • Pros • Autoclavable • Even pathogen delivery • Nose piece • Split Tubing • Cons • Too difficult to manufacture • Nose piece • Tubing • Too expensive • Infects only 4 animals at once

  14. Alternative Design 3: • Pros • Jars for various animal sizes • Latex diaphragms for restraint • Circulation fan for distribution • Fits under fume hood • Cons • Too difficult to manufacture • Too expensive

  15. Lid with Gasket Tape and Output Valve • PVC Pipe Stand • Stainless Steel Mesh Shelf • Input Valve with Elbow for Air Flow • Latches

  16. System Advantages • Fully Autoclavable • Air Tight System • Increased Efficiency • Increased Safety • Even Distribution of Pathogen • Increased Holding Container Size • Small Animals to Large Animals • Greater Numbers per Trial

  17. Attached Clamp Manufacturing: Clamp Attachment Clamp Assembly • Holes Drilled at Equal Intervals • Holes Filed and Finished to Remove Sharp Edges • Teflon Gasket Custom Fit to Screws • Screws, Teflon Washer, Nut, and Clamp Attached Nut Screw Teflon Washer

  18. Manufacturing: Lid • Lid • 3 layers of PTFE gasket tape on inside lip • Creates seal between pot and lid

  19. Inflow Elbow Inflow Hose Nipple Manufacturing: Inflow • ½” Diameter Hole Drilled with Step Drill • Elbow Positioned on Threaded End of Hose Nipple • Hose Nipple Welded to Pot • Elbow positioned to 45o to promote air circulation

  20. Outflow Hose Nipple Manufacturing: Outflow • ½” Diameter Hole Drilled with Step Drill • Hose Nipple Welded to Pot

  21. Cost • 48 qt. Stainless Steel Stockpot $90.00 • Draw Latches (8) $33.40 • Woven Wire Cloth $33.00 • Stainless Hose Adapters (2) $16.50 • 45o Elbow Fitting $11.60 • PTFE Gasket Tape $32.50 • Welding and Labor $50.00 TOTAL $267.00

  22. Design Matrix Design1Design2Design3Final Cost 4 3 2 9 Ease of 5 6 3 7 Manufacture Cleaning 5 4 5 8 Safety 7 6 7 7 Overall 211917 31

  23. FUTURE WORK TESTING Test with water and non-pathenogenic bacteria Efficiency Consistency Safety Air Flow Improved animal holding chambers Nose/Head only

  24. References • www.McMaster-Carr.com • www.EastmanOutdoors.com • www.glascol.com/products/biotimag/biotech.html • www.hc-sc.gc.ca/pphb-dgspsp/publicat/lbg-ldmbl-96/ • www.nalgene.com

  25. Special Thanks • Professor Tompkins (Advisor) • Orrie Lokken (ME Shop Supervisor) • Adel Talaat (Client) • Nick Anders (our fallen leader)

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