VeterinaryManagementChallengesandFutureDirections

Including

TechnicalConsiderationsforImagingandSurgery
inGnotobioticAnimals
BettyR.Theriault,DVM,DACLAM
AssociateProfessor,DepartmentofSurgery
&AnimalResourcesCenter
TheUniversityofChicago

December20,2016

VeterinaryManagementChallenges
(Opportunities)
DevelopingAGermFreeorGnotobioticProgram
inthecontextofaSpecificPathogenFreeEra

Education,Training& Communication

GnotoMottostoLiveBy
Ifitwereeasy,everyonewouldbedoingit
Before2005veryfewwere
Between2005and2010,veryfewadditional
Currentlyeveryonewouldliketo

Nothingisaseasyasitappears
Everythingwilltakelongerthananticipated

BestPracticeConsiderations

Food,WaterandBedding
IsolatorandCagingSupplies

Theharshrealityoftherelativecostofmaintaininggnotobiotes
versustheclassicapproachofanopenroomsoonmadeitreadilyapparent
thatthiswasnotpractical tosaynothingoftheaddedinconvenience
ofhavingtomanipulateanimalsandcagesinsideaplasticbubble.
RogerP.Orcutt

HealthStatusMonitoring
AdventitiousPathogens In
SpecificPathogenFree(SPF)Colonies

SterilityIn
GermFreeColonies

Frequency?

Frequency?

Methodology?

Methodology?

LiveSentinel
SoiledBedding
Contact

EnvironmentalPCR
Direct(Animal)
Indirect(Cage,Plenum)

Who?

Cultivation
16sPCR

Who?

Developingand/orAdapting
AnimalModelsto
MicrobiomeResearchNeeds

InstitutionalEducation&Training
Thepurposeofquarantinepracticesis
containmentofpotentialadventitiouspathogens
topreventthemfromenteringestablishedcolonies
&
Thepurposeofstandardcarepracticesisthe
protectionoftheSpecificPathogenFreestatusof
establishedcolonies
&
Thepurposeofgnotobioticpracticesisexclusionofall
detectable,nondefinedorganisms

Howdoweaccomplishalloftheaboveinthecontextof
GermFree,Gnotobiotic,DefinedFlora,SPFandBiohazardousStatus
whentheymayalloverlapinmicrobiomeresearch...?

Education,TrainingandCommunication
AnimalCareandUseProgramPersonnel
ResearchGroup
InstitutionalAnimalCareandUseCommittee
Veterinarian,Scientist,NonScientist,PublicMember

InstitutionalBiosafetyCommittee
Veterinarian,BiosafetyOfficer,Scientist,InfectiousDisease
Physician,NonScientist,PublicMember

Programvisitors

AdvancedStudyInterventions
TechnicalConsiderations
Manyproceduresinveterinarypracticerequire
chemicalimmobilizationoranesthesia
ImagingModalities
Radiotherapy
AnimalModelsrequiringadministrationoftestagents
(speciesandadministrationroutedependent)

Collectionoftestspecimensand/ordiagnostics
Blood,Urine,Feces,BronchialveolarLavage,
TissueBiopsy,PhysiologicData

Anesthesia
DirectChallenges

IndirectChallenges

Species
Breed
Strain
Gender
Age
Condition

ProtocolReview
Institutionalguidanceon
doseranges
Literatureguidanceondose
ranges
Pilottrialsforanesthesia
doseswithinstudydesign

GermFree
ExGermFree
HumanizedMicrobiome

MicrobiomeInfluenceson
AnesthesiaandAnalgesia
Mightvariationsincompositionanddiversityofthe
microbiomeinfluencemechanismsofhomeostasis
inthehealthand/ordiseasestateofanorganism?
Mightthemicrobiomealsoinfluencehowthe
organismrespondstoagentsdependentuponthesame
mechanismsfortherapeuticeffect?

ReceptorFunction
EnzymeFunction
HepaticClearance
IntestinalClearance
UrinaryExcretion

RespiratoryPhysiology
DosingIntervals
DosingRoutes

AnesthesiaandAnalgesia
MaintenanceChallenges
DrugActivity
InjectableAgents
Ketamine
Xylazine
Yohimbine
Acepromazine
Dexmetatomidine
Atipamizole
Buprenorphine
NSAIDs
InhalantAnesthetics
Isoflurane
Sevoflurane

PatientSupport

Monitoring
Directaccesstoassessment
DirectaccesstoIntervention
Addedriskforcontamination
Abilitytosterilizesupport
equipment

Anesthesia
ProceduralChallenges
Immobilization
Imaging
OpticalImaging
Ultrasound
MRI/CT/PET/SPECT
DEXAScan
(DualEnergyXrayAbsorptiometry)

SurgicalProcedures

TimetoInduction
DurationofEffect
Redosing
RecoveryTime

TreatmentInterventions
Irradiation
Measurements
TestAgentDosing

PLANNINGISKEY

ModelChallenges
Bioexclusion

Biocontainment

EquipmentInnovations
TrexlerChallenge:
WhentaskedbyaResearcher:
HowdoIgetmytestequipmentintoanisolator?
Trexlerwouldrespondby:
HowdoIwrapmybubblearoundyourtestequipment?

Examplesofearlymodelsofflexiblefilmisolators
introducedbyPhilipTrexlerattheLOBUNDInstitute.
TheLaboratoryRat,Vol.22006.

Procedural&
EquipmentChallenges

HowdoIwrapabubblearoundmyinvestigatorsanimal?

ValidationofanAnimalIsolationImagingChamberforUsein
AnimalBiosafetyLevel3Containment
Alderman,T.S.,FrothinghamR.,andSempowski,G.D.,
ApplBiosaf.2010;15(2):6266.

SurgicalChallenges

Surgerywithout
SurgicalIsolatorProtection
Preparation of Surgical Field
Preparation of ALL materials
Transfer of animal from and to sterile housing
Preparation of Surgeon with Garb
Refine surgical technique
Anesthesia and Analgesia for animal
Patient recovery

LongtermMaintenanceofSterilityAfterSkinTransplantationinGermfreeMice
Theriault,B.,Wang,Y.,Chen,L.,Vest,A.,Bartman,C.,Alegre,ML.
TransplantationDirect2015;1.Publishedonline17September2015

ChallengesWithinaProgram
FacilityDevelopmentandDesign
Staffing/Training/Expertise
InstitutionalExperience
IACUC
IBC
Veterinarians
Investigator
HealthStatusMonitoring
MicrobiomeinAnesthesia AgentMetabolismandEffect
Accesstopreciousandnovelresources
FinancialChallenges
CollaborativeStudies

Challenging
TraditionalBoundaries

CollaborativeStudies
InterInstitutionalMemorandumofUnderstanding(MOU)
MaterialTransferAgreement(MTA)
CostofFacilitiesandOperations
Costofspecializedequipment
CostandRiskofTransport
FacilitatingCollaborations(WithinandBetweenInstitutions)
TrainingandUnderstandingofSterileTechnique
CentersofExpertiseandExcellence

Acknowledgements
UniversityofChicagoInvestigators

UniversityofChicagoAnimalResourcesCenter

EugeneChang
VanessaLeone
JoePierre
KarenHo
CathrynNagler
*AndrewStefka
*TaylorFeehley
MarisaAlegre
YingWang
*CarolineBartman
JohnAlverdy
*MarkWard
EricaClaud
RalphWeichselbaum
YuanZhang
CaroleOber
TatyanaGolovkina
AlexanderChervonsky

GeorgeLangan Director
CarolMathieu
ChrisWhaley
KeithFournier
GnotobioticResearchAnimalFacility(GRAF)

Collaborators
UniversityofArizona Tucson
DonataVercelli
JustynaGozdz
NorthShoreHealthcareSystem
JingLu
AlexanderDrobyshevsky

*denotes individuals who have left the institution

Staff (CharlesRiver)
ChristinaOlivares Supervisor
*TracyLatalladi
*NicoleWest
*EdytaNovak
*ScottBlameuser
*AlanVest
KristenBrockway
*LoveOwens
CarrieHoffman
MichaelYabes
*MeganBales
KristinKolar
*AndreaCrawford
*HaleyMontes
NicoleMartinec
MelanieSpedale
DerrickSummerville
StanRoss
RudolphCooper
StevePaynter ProjectManager