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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 Department of Animal and Biomedical Sciences 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. 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 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 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 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 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 Alternative Design 1: Pros – Dual Chamber System – Split Air Flow – Latex Flange around Neck Air input Cons – Specific only to mice – Too difficult to manufacture – Too expensive animal Air output Animal head Air input 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 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 •Lid with Gasket Tape and Output Valve •PVC Pipe Stand •Stainless Steel Mesh Shelf •Input Valve with Elbow for Air Flow •Latches 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 Manufacturing: Clamp Attachment Nut 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 Screw Teflon Washer Attached Clamp Manufacturing: Lid Lid – 3 layers of PTFE gasket tape on inside lip – Creates seal between pot and lid Manufacturing: Inflow Inflow Elbow Inflow Hose Nipple 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 Manufacturing: Outflow Outflow Hose Nipple ½” Diameter Hole Drilled with Step Drill Hose Nipple Welded to Pot Cost 48 qt. Stainless Steel Stockpot Draw Latches (8) Woven Wire Cloth Stainless Hose Adapters (2) 45o Elbow Fitting PTFE Gasket Tape Welding and Labor TOTAL $90.00 $33.40 $33.00 $16.50 $11.60 $32.50 $50.00 $267.00 Design Matrix Design1 4 Design2 3 Design3 2 Final 9 Ease of Manufacture 5 6 3 7 Cleaning 5 4 5 8 Safety 7 6 7 7 Overall 21 19 17 Cost 31 FUTURE WORK TESTING –Test with water and non-pathenogenic bacteria Efficiency Consistency Safety Air Flow Improved animal holding chambers –Nose/Head only 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 Special Thanks Professor Tompkins (Advisor) Orrie Lokken (ME Shop Supervisor) Adel Talaat (Client) Nick Anders (our fallen leader)