1. BSL3 To Infect

2. InhalationLaboratory

3. SystemAnimals

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

8. Background Client’s goals of animal infection study Client’s goals of animal infection study –To study the pathenogenesis of tuberculosis (BSL3) –To develop a cure for TB patients Safety Precautions 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 Main chamber: 75 cm x 50 cm x 15 cm Inner chambers: radius 8 cm, height 12 cm Inner chambers: radius 8 cm, height 12 cm Nebulizer, Ethanol Trap, HEPA Filter Nebulizer, Ethanol Trap, HEPA Filter Tupperware 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: Dimensions: –Length: 25 cm – 75 cm –Width: 40 cm – 30 cm –Height: 10 cm – 70 cm Withstand the Autoclaving Process Withstand the Autoclaving Process Storage at Room Temperature Storage at Room Temperature Air Tight Structure Air Tight Structure Deliver 5 mL of culture in 20 minutes Deliver 5 mL of culture in 20 minutes Service Life: 50 to 100 Uses Service Life: 50 to 100 Uses

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

13. Alternative Design 2: Pros Pros –Autoclavable –Even pathogen delivery  Nose piece  Split Tubing Cons 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

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

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

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

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

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

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

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

23. 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 21 19 17 31

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

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

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