BSL3BSL3

To InfectTo Infect

InhalationInhalation

LaboratoryLaboratory

SystemSystem

AnimalsAnimals

Team MembersTeam MembersNick Anders – Team LeaderNick Anders – Team Leader

Stacey Hoebel – Stacey Hoebel – CommunicationsCommunications

Jeff Swift – BWIGJeff Swift – BWIG

Maja Middleton – BSACMaja Middleton – BSAC

Kayla Ericson – BWIGKayla Ericson – BWIG

Austin Ramme - BWIGAustin Ramme - BWIG

AdvisorAdvisorProfessor Willis TompkinsProfessor Willis Tompkins

ClientClientAdel TalaatAdel Talaat

Assistant ProfessorAssistant Professor

Department of Animal and Biomedical SciencesDepartment of Animal and Biomedical Sciences

AbstractAbstract

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

Problem Problem StatementStatement

Develop a safe inhalation system for Develop a safe inhalation system for animalanimal

infectioninfectionShould be able to infect animals Should be able to infect animals ranging inranging in

size from mice, rats, and guinea pigs size from mice, rats, and guinea pigs to to

rabbitsrabbitsShould be equipped with pathogen Should be equipped with pathogen escapeescape

prevention measures to insure safety prevention measures to insure safety in thein the

surrounding environmentsurrounding environment

BackgroundBackground

Client’s goals of animal infection studyClient’s goals of animal infection study– To study the pathenogenesis of To study the pathenogenesis of

tuberculosis (BSL3)tuberculosis (BSL3)– To develop a cure for TB patientsTo develop a cure for TB patients

Safety PrecautionsSafety Precautions– Proper training and protective clothingProper training and protective clothing– Non-porous materialNon-porous material– Autoclave in work zoneAutoclave in work zone– HEPA filtrationHEPA filtration– Sealed exhaust system with directional Sealed exhaust system with directional

airflow airflow

Current Device in UseCurrent Device in Use

Main chamber: 75 cm x 50 cm x 15 cmMain chamber: 75 cm x 50 cm x 15 cm Inner chambers: radius 8 cm, height 12 cmInner chambers: radius 8 cm, height 12 cm Nebulizer, Ethanol Trap, HEPA FilterNebulizer, Ethanol Trap, HEPA Filter TupperwareTupperware

Current Models availableCurrent 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 SpecificationsDesign Specifications Dimensions:Dimensions:

– Length: 25 cm – 75 cmLength: 25 cm – 75 cm– Width: 40 cm – 30 cmWidth: 40 cm – 30 cm– Height: 10 cm – 70 cmHeight: 10 cm – 70 cm

Withstand the Autoclaving ProcessWithstand the Autoclaving Process Storage at Room TemperatureStorage at Room Temperature Air Tight StructureAir Tight Structure Deliver 5 mL of culture in 20 minutesDeliver 5 mL of culture in 20 minutes Service Life: 50 to 100 UsesService Life: 50 to 100 Uses

Alternative Design 1:Alternative Design 1:

ProsPros– Dual Chamber Dual Chamber

SystemSystem– Split Air FlowSplit Air Flow– Latex Flange around Latex Flange around

NeckNeck ConsCons

– Specific only to miceSpecific only to mice– Too difficult to Too difficult to

manufacturemanufacture– Too expensiveToo expensive

Air input

animal

Air input

Animal head

Air output

Alternative Design 2:Alternative Design 2: ProsPros

– AutoclavableAutoclavable– Even pathogen deliveryEven pathogen delivery

Nose pieceNose piece Split TubingSplit Tubing

ConsCons– Too difficult to Too difficult to

manufacturemanufacture Nose pieceNose piece TubingTubing

– Too expensive Too expensive – Infects only 4 animals Infects only 4 animals

at onceat 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 AdvantagesSystem Advantages

Fully AutoclavableFully Autoclavable

Air Tight SystemAir Tight System– Increased EfficiencyIncreased Efficiency– Increased SafetyIncreased Safety

Even Distribution of PathogenEven Distribution of Pathogen

Increased Holding Container Increased Holding Container SizeSize– Small Animals to Large AnimalsSmall Animals to Large Animals– Greater Numbers per TrialGreater Numbers per Trial

Manufacturing: Clamp Manufacturing: Clamp AttachmentAttachment

Holes Drilled at Holes Drilled at Equal IntervalsEqual Intervals

Holes Filed and Holes Filed and Finished to Remove Finished to Remove Sharp EdgesSharp Edges

Teflon Gasket Teflon Gasket Custom Fit to Custom Fit to ScrewsScrews

Screws, Teflon Screws, Teflon Washer, Nut, and Washer, Nut, and Clamp AttachedClamp Attached

Attached Clamp

Teflon Washer

Clamp AssemblyNut

Screw

Manufacturing: LidManufacturing: Lid

LidLid– 3 layers of PTFE gasket tape on inside 3 layers of PTFE gasket tape on inside

liplip– Creates seal between pot and lidCreates seal between pot and lid

Manufacturing: InflowManufacturing: Inflow

½” Diameter Hole ½” Diameter Hole Drilled with Step Drilled with Step DrillDrill

Elbow Positioned Elbow Positioned on Threaded End of on Threaded End of Hose Nipple Hose Nipple

Hose Nipple Hose Nipple Welded to PotWelded to Pot

Elbow positioned to Elbow positioned to 4545oo to promote air to promote air circulationcirculation

Inflow Elbow

Inflow Hose Nipple

Manufacturing: OutflowManufacturing: Outflow

½” Diameter Hole ½” Diameter Hole Drilled with Step Drilled with Step DrillDrill

Hose Nipple Hose Nipple Welded to PotWelded to Pot

Outflow Hose Nipple

CostCost 48 qt. Stainless Steel Stockpot 48 qt. Stainless Steel Stockpot

$90.00$90.00 Draw Latches (8)Draw Latches (8) $33.40$33.40 Woven Wire ClothWoven Wire Cloth $33.00$33.00 Stainless Hose Adapters (2)Stainless Hose Adapters (2) $16.50$16.50 4545oo Elbow Fitting Elbow Fitting $11.60$11.60 PTFE Gasket TapePTFE Gasket Tape $32.50$32.50 Welding and Labor Welding and Labor $50.00$50.00

TOTALTOTAL $267.00$267.00

Design MatrixDesign MatrixDesign1Design1 Design2Design2 Design3Design3 FinalFinal

CostCost 4 4 3 3 2 2 9 9

Ease of Ease of 5 5 6 6 3 3 7 7

ManufactureManufacture

CleaningCleaning 5 5 4 4 5 5 8 8

SafetySafety 7 7 6 6 7 7 7 7

Overall Overall 2121 1919 1717 31 31

FUTURE WORKFUTURE WORKTESTINGTESTING

–Test with water and non-pathenogenic Test with water and non-pathenogenic bacteriabacteria

EfficiencyEfficiencyConsistencyConsistencySafetySafetyAir FlowAir Flow

Improved animal holding chambersImproved animal holding chambers–Nose/Head onlyNose/Head only

ReferencesReferences

www.McMaster-Carr.comwww.McMaster-Carr.com

www.EastmanOutdoors.comwww.EastmanOutdoors.com

www.glascol.com/products/biotimag/biotech.htmlwww.glascol.com/products/biotimag/biotech.html

www.hc-sc.gc.ca/pphb-dgspsp/publicat/lbg-ldmbl-www.hc-sc.gc.ca/pphb-dgspsp/publicat/lbg-ldmbl-

96/96/

www.nalgene.comwww.nalgene.com

Special ThanksSpecial Thanks

Professor Tompkins Professor Tompkins (Advisor)(Advisor)

Orrie Lokken (ME Orrie Lokken (ME Shop Supervisor)Shop Supervisor)

Adel Talaat (Client)Adel Talaat (Client)

Nick Anders (our Nick Anders (our fallen leader)fallen leader)