2014  District  5  AALAS  Meeting  

THURSDAY,  MAY  15    

TRACK  1  PRESENTATION  ABSTRACTS  RNA  Interference  Therapy  for  Huntington’s  Disease.  Richard  Grondin,  Ph.D.,  Department  of  Anatomy  and  Neurobiology,  University  of  Kentucky,  Lexington,  Kentucky    Huntington’s  disease   (HD)   is   a   fatal  neurodegenerative  disease  caused  by  expression  of  a  mutant  huntingtin   (Htt)  protein  containing   an   expanded   polyglutamine   repeat   region.   Consequently,   lowering   Htt   expression   is   regarded   as   a   promising  approach  to  treat  HD  patients.  Both  mutant  and  wildtype  Htt  can  be  reduced  in  vivo  via  ribonucleic  acid  (RNA)  interference  using  synthetic,  double-­‐stranded  small  interfering  RNAs  (siRNAs).  Whether  therapies  that  reduce  both  wildtype  and  mutant  Htt  will  be  beneficial  or  harmful  to  patients  depends  in  part  on  whether  partial  reduction  of  wildtype  Htt  can  be  tolerated  in  the  adult  primate  brain.        The   present   series   of   studies   was   conducted   to   assess   the   effects   of   lowering   wildtype   Htt   expression   with   continuous  delivery   of   siRNAs   targeting   Htt   into   the   adult   nonhuman   primate   brain.   Abdominally   implanted   programmable   infusion  pumps   connected   to   brain   implanted   catheters   were   used   to   continuously   infuse   siRNAs   into   the   putamen   of   rhesus  macaques.  In  a  first  series  of  experiments,  vehicle  or  varying  concentrations  of  14C-­‐siRNAs  were  infused  for  7  days  at  a  flow  rate   of   0.3   µL/min,   consistent  with   convection   enhanced   delivery.   In   a   second   series   of   experiments,   vehicle   or   a   single  concentration  of  14C-­‐siRNAs  was  infused  at  the  same  flow  rate  for  varying  period  of  times,  ranging  from  1  to  7  days.  This  was  followed  by  treatment  cessation  for  10  or  24  days  in  selected  animals.    Substantial  knockdown  of  Htt  mRNA,  up  to  45%,  was  observed  throughout  the  putamen.  The  onset  of  Htt  suppression  occurred  rapidly,  peaking  within  about  3-­‐4  days  of  infusion  initiation  and  slowly  declining  overtime  at  a  rate  of  about  1.5%  per  day.  Other  than  circumscribed  tissue  responses  around  the  catheter  tip,  there  was  no  cell  loss  in  the  putamen.  Our  data  support  that  intraparenchymal  delivery  of  siRNAs  designed  to  lower  both  wildtype  and  mutant  Htt  has  significant  clinical  potential.        Husbandry,  Veterinary  Support  and  Common  Clinical  Problems  of  Brain  Cannulated  NHP  models.  Jeffrey  Smiley,  D.V.M.,  Ph.D.,  D.A.C.L.A.M.,  Division  of  Laboratory  Animal  Resources,  University  of  Kentucky,  Lexington,  Kentucky    Dr  Smiley  will  give  a  short  presentation  on  husbandry  considerations  and  veterinary  medical  care  and  support  of  for  brain  cannulated  NHP  models.  Topics  of  discussion  will  include  pre-­‐study  evaluation  of  candidate  animals,  quarantine  procedures,  animal  husbandry  considerations  and  common  medical  problems.    

Epicardial  Cell  Therapy  for  Heart  Disease  in  Rats:  the  Science,  Surgery,  and  Related  Animal  Care.  Allison  L.  Aird,  B.S.  and  Amanda  Jo  LeBlanc,  Ph.D.,  Cardiovascular  Innovation  Institute,  University  of  Louisville  and  Jewish  Hospital,  Louisville,  Kentucky    In  aged  rats  and  humans,  the  ability  of  the  heart  to  increase  blood  flow  when  stimulated  is  significantly  decreased  and  may  contribute  to  the  increased  risk  for  cardiovascular  disease  as  people  age.  Our  laboratory  has  isolated  a  cell  population  from  adipose  tissue  to  create  a  therapeutic  cell  sheet  for  epicardial  implantation.  Following  implantation  of  the  cell  sheet,  we  have  successfully  treated  ischemia  following  myocardial  infarction  and  also  improved  blood  flow  in  hearts  from  aged  rats.  We  will  discuss  surgical  procedures  for  myocardial  infarction  and  cell  sheet  implantation  in  rats,  post-­‐surgical  care,  and  special  considerations  for  treating  an  aged  population  of  rodents.        Research  in  an  ABSL3  Environment:    Special  Considerations  from  Different  Perspectives.  Clarissa  Cowan1,4,  Jonathan  Warawa,  Ph.D.2,4,  Karen  Powell,  D.V.M.,  Ph.D.3,4,  Carol  Vanover,  R.L.A.T.3,4,  Jennifer  Kraenzle,  R.L.A.T.G.3,4,  1Department  of  Environmental  Health  and  Safety,  2Center  for  Predictive  Medicine,  3Research  Resources  Facilities,  4Regional  Biocontainment  Laboratory,  University  of  Louisville,  Louisville,  Kentucky    The  Regional  Biocontainment  Laboratory  (RBL)  at  the  University  of  Louisville  is  one  of  only  11  sites  funded  by  NIH  for  the  secure  biosafety  research  of  BSL3  agents.  The  mission  of  the  RBL  is  to  improve  human  health  through  the  development  of  vaccines  and  therapeutics  for  the  prevention  and/or  treatment  of  infectious  diseases  that  could  pose  serious  threat  or  be  used  in  acts  of  bioterrorism.    

2014  District  5  AALAS  Meeting  

Research  of  this  nature  entails  special  considerations  at  all  levels.  This  seminar  will  provide  an  insider’s  view  of  how  a  team  consisting  of  a  safety  and  compliance  specialist;  a  researcher;  vivarium  manager;  husbandry  technicians;  and  vet  staff  work  together  to  address  the  unique  challenges  encountered  in  a  BSL3  environment.    Understanding  and  Enhancing  Motor  Performance  Following  Spinal  Cord  Injury:    Animal  Models  to  Human  Subjects  Research.  Dena  Howland,  Ph.D.,  O.T.1,2,3,4,5,  Wilbur  O’Steen1,2,3,  Karen  Powell,  D.V.M.,  Ph.D.6,  Aaron  Rising,  Ph.D.1,2,3,  Kimberly  Cheffer,  B.A.1,2,3,  Kyle  Whyland,  B.A.1,2,3,  S.A.  Trimble,  M.S.,  P.T.7,,  Andrea  Behrman,  Ph.D.,  P.T.2,3,  1Robley  Rex  VAMC,  Louisville,  KY,  2Kentucky  Spinal  Cord  Injury  Research  Center,  3Department  of  Neurological  Surgery,  4Anatomical  Sciences  and  Neurobiology,  5Department  of  Bioengineering,  and  6Research  Resources  Facilities,  University  of  Louisville,  and  7Frazier  Rehabilitation  Institute,  Louisville,  Kentucky    Our  extended  and  collaborative  research  group  focuses  on  understanding  the  response  of  the  spinal  cord  to  injury  and  identifying  approaches  to  enhance  repair,  plasticity  and  motor  recovery.    We  employ  a  bi-­‐directional  research  approach  in  which  results  from  basic  science  studies  are  used  to  direct  and  contribute  to  the  advancement  of  human  clinical  research  and  treatment  (translation),  and  where  findings  in  clinical  research  and  practice  are  used  to  direct  questions  and  refine  hypotheses  that  are  most  suitable  for  inquiry  in  the  basic  science  domain  (reverse  translational  research).    Thus,  the  goal  is  an  ongoing  dialogue  between  basic  science,  human  subject  research  and  the  clinic  to  achieve  cutting  edge  research  and  treatment.      The  cat  has  contributed  significantly  to  our  understanding  of  motor  control  and  serves  as  an  invaluable  model  for  aspects  of  our  translational  spinal  cord  research.  In  addition  to  cellular  and  molecular  assessments  to  understand  changes  in  protein  expression,  spinal  circuitry  and  long  tracks,  we  evaluate  functional  changes  using  a  variety  of  approaches  which  range  from  assessment  of  basic  reflexes  to  3D  kinematic  assessments  of  locomotion  which  parallel  those  used  in  our  human  subjects  research.    At  the  clinical  research  level,  our  major  focus  currently  is  on  enhancing  motor  recovery  using  activity-­‐based  interventions.    In  particular,  we  are  using  Locomotor  Training  (LT)  in  young  children  with  severe,  chronic  spinal  cord  injuries  who  show  little  to  no  clinical  potential  for  walking  recovery.    Our  ongoing  work  suggests  that  up  to  50%  of  these  children  may  develop  walking  with  this  approach  and  that  all  show  benefits  through  improvements  in  trunk  control  critical  for  sitting  posture  and  upper  extremity  use.    Among  our  goals  are  to  develop  predictive  measures  to  identify  those  with  the  greatest  potential  to  respond  to  LT,  develop  approaches  to  accelerate  responses  to  LT,  and  to  understand  underlying  circuitry  changes.  Support:  Dept  of  VA  (RR&D),  Craig  H.  Neilsen  Foundation,  NINDS,  NICHD,  Christopher  and  Dana  Reeve  Foundation,  Frazier  Rehab  Institute  and  Kentucky  One  Health,  Rebecca  F.  Hammond  Trust,  Kentucky  Spinal  Cord  and  Head  Injury  Trust,  Commonwealth  of  Kentucky  Challenge  for  Excellence,  Kosair  Charlities    Bench  to  Bedside  Approaches  for  the  Treatment  of  Traumatic  Brain  Injury.  Patrick  Sullivan,  Ph.D.,  and  Andrea  Sebastian,  B.S.,  Spinal  Cord  and  Brain  Injury  Research  Center,  University  of  Kentucky,  Lexington,  Kentucky    Traumatic  brain  injury  (TBI)   is  a  devastating  healthcare  problem  in  the  United  States  with  an  estimated  1.7  million  injuries  annually  at  an  estimated  yearly  cost  of  greater   than  76.5  billion  dollars.  However,   there  are  currently  no  pharmacological  treatments   approved   for   the   clinical   treatment   of   this   condition.   Compelling   experimental   data   demonstrates   that  mitochondrial  dysfunction   is   a  pivotal   link   in   the  neuropathological   sequalae  of  brain   injury.   In   support  of   this  hypothesis  several   reports   have   demonstrated   that   inhibition   of   the   mitochondrial   permeability   transition   pore   with   the  immunosuppressant  drug  cyclosporin  A  (CsA)  is  efficacious.  Accordingly,  CsA  is  being  moved  forward  into  late  stage  clinical  trials  for  the  treatment  of  moderate  and  severe  TBI.  However,  several  unknowns  exist  concerning  the  optimal  therapeutic  window  for  administering  CsA  at  the  proposed  dosages  to  be  used  in  human  studies.  The  present  study  utilized  a  moderate  (1.75  mm)  unilateral  controlled  cortical  impact  model  of  TBI  in  order  to  determine  the  most  efficacious  therapeutic  window  for  initiating  CsA  therapy,  rats  were  administered  an  i.p.  dose  of  CsA  (20mg/kg)  or  vehicle  at  1,  3,  4,  5,  6,  8  hrs  post-­‐injury.  Together   these   results   illustrate   the   importance   of   initiating   therapeutic   interventions   such   as   CsA   as   soon   as   possible  following   TBI,   preferably  within   4   hrs   post-­‐injury   to   achieve   the   best   possible   neuroprotective   effect.   However,   the   drug  appears  to  retain  some  protective  efficacy  even  when  initiated  as  late  as  8  hrs.

   

2014  District  5  AALAS  Meeting  

Cell-­‐Based  Therapy  for  Vascularized  Composite  Allografts.  James  B.  Hoying,  Ph.D.  Robert  M.  Reed,  Jacob  R.  Dale,  Jeremy  S.  Touroo,  Christina  L.  Kaufman,  and  Stuart  K.  Williams,  Ph.D.,  Cardiovascular  Innovation  Institute  and  Christine  M.  Kleinert  Research  Institute,  University  of  Louisville,  Louisville,  Kentucky    Transplantation  of   vascularized   composite   allografts   (VCA),   such   as   hand   and   face   transplants,   entail   a   number  of   clinical  challenges.   In   addition   to   the   serious   complications   arising   from   immune   rejection,   the   health   of   the   allografts   can   be  compromised   by   graft-­‐related   vasculopathies.   Adipose-­‐derived   stromal   vascular   fraction   (aSVF)   cells   have   proven   anti-­‐inflammatory  and  pro-­‐vascular  as  well  as  suspected  immune  modulatory  activities.  With  this  in  mind,  we  are  exploring  the  therapeutic  potential  of  aSVF  cells  in  improving  VCA  outcomes.  Using  a  rat  model  of  VCA  involving  the  transplantation  of  the  intact   lower   hindlimb   of   a   Brown   Norway   rat   to   a   Lewis   rat   via   a   femoral   artery/vein   pedicle,   we   are   delivering   freshly  isolated  aSVF  cells  via  multiple  intra-­‐graft  injections  either  during  or  after  a  2  week    regimen   of   the   immunosuppressant   tacrolimus   post-­‐transplantation.   The   aSVF   cells   are   harvested   from   a   third   donor   rat  syngeneic  with  the  Lewis  graft   recipient.  Analysis  of  allograft  health   included  assessment  of  clinical  signs  of  skin  rejection,  laser   Doppler   perfusion   imaging,   X-­‐ray   radiography,   splenocyte   flow   cytometry,   and   histology   of   graft   tissues.   This   pre-­‐clinical   model   is   proving   very   useful   in   the   development   of   a   cell-­‐based   therapy   for   augmenting   vascularized   composite  allografting.    

 

THURSDAY,  MAY  15  

TRACK  2  PRESENTATION  ABSTRACTS  

The  Future  of  Mouse  Embryo  Transfer:  Achieving  the  3Rs  with  the  NSET  Device.  Barbara  Stone,  Ph.D.,  ParaTechs  Corporation,  Lexington,  Kentucky    Non-­‐surgical  transfer  techniques  for  mouse  embryos  and  sperm  provide  animal  welfare  benefits  for  assisted  reproduction  of  mice.    While  surgical  embryo  transfer  (ET)  is  an  effective  method  for  transfer  of  embryos  into  the  uterine  horn  of  mice,  surgery  is  expensive,  time  consuming,  and  requires  technical  expertise.    Surgery  is  also  a  stressful  procedure  for  the  mouse,  which  has  to  be  anesthetized  and  treated  with  an  analgesic.  We  have  developed  a  simple,  brief  procedure  for  ET  using  a  non-­‐surgical  embryo  transfer  (NSET)  device.  Once  embryos  are  loaded  into  the  NSET  device,  the  tapered  NSET  catheter  passes  through  the  vagina  and  traverses  the  cervix  to  deposit  the  embryos  into  the  uterine  horn  of  a  recipient  mouse.    Since  the  NSET  procedure  does  not  require  sedation,  opening  of  the  inner  body  cavity,  or  use  of  an  analgesic,  this  procedure  is  less  stressful  for  the  mouse  than  surgery.    In  proof,  our  results  show  that  when  pseudopregnant  mice  are  monitored  by  electrocardiography,  the  non-­‐surgical  procedure  does  not  affect  heart  rate.      The  NSET  procedure  also  has  no  effect  on  the  level  of  the  stress  biomarker  fecal  corticosterone  measured  by  ELISA  analysis.    However,  surgery  or  anesthesia  does  lower  heart  rate  and  increases  levels  of  fecal  corticosterone.    Additional  studies  show  that  the  NSET  procedure  is  effective  for  transfer  of  mouse  embryos  for  the  purposes  of  rederivation,  cryorecovery,  and  transgenic  research.    Embryos  can  be  successfully  transferred  with  the  NSET  technique  after  cryopreservation,  in  vitro  fertilization  (IVF),  or  embryonic  stem  (ES)  cell  injection  as  an  alternative  to  traditional  surgical  embryo  transfer.    The  NSET  device  has  also  successfully  transferred  mouse  sperm  for  artificial  insemination.  Therefore,  the  NSET  technique  is  an  example  of  a  3Rs  refinement,  an  alternative  to  a  surgical  procedure,  which  will  lead  the  way  for  improved  assisted  reproductive  techniques  in  mice.      

Pieces  of  the  Puzzle:  Research  and  You  Coming  Together.  Jefferson  Childs,  L.V.T.,  R.L.A.T.G.,  C.M.A.R.,  S.R.S.,  I.L.A.M.,  Cincinnati  Children’s  Hospital,  Cincinnati,  Ohio    Case  studies  in  humans  will  be  discussed  that  illustrate  how  the  research  that  we  do  affects  the  outcome  and  diagnosis  of  uncommon  disorders.  Research  all  over  the  country  can  come  together  on  cases  to  help  human  patients  in  hospitals  anywhere.  Case  one  is  a  Periodic  Fever  Syndrome;  Case  two  is  a  Pectus  Excavatum  case.  Not  only  are  patients  with  cancer  and  heart  disease  affected  by  research  but  also  that  even  basic  research  can  help  cure  medical  conditions.  Everyone  in  this  field  has  a  story  to  tell  of  how  the  work  we  do  connects  use  to  successes  in  medical  care,  and  we  should  be  proud  of  what  we  do  and  why  we  do  it.  We  do  not  do  anything  that  needs  to  be  hidden  from  the  public.  Each  and  every  one  of  us  contribute  to  medical  breakthroughs  on  a  daily  basis  whether  they  take  place  at  our  institution  or  across  the  country.  

2014  District  5  AALAS  Meeting  

A  Strong  Biosecurity  Program  Supports  Strong  Animal  Welfare.  William  Porter,  D.V.M.,  D.A.C.L.A.M,  and  Mandy  J.  Horn,  M.S.,  Harlan  Laboratories,  Inc.,  Indianapolis,  Indiana    Biosecurity,  as  mentioned  in  the  current  “Guide  for  the  Care  and  Use  of  Laboratory  Animals”,  encompasses  all  measures  taken  to  address  known  or  unknown  infections  in  an  animal  colony.    In  laboratory  rodent  colonies,  biosecurity  is  most  critical  as  large  numbers  of  animals  are  usually  housed  in  close  proximity  to  one  another.    Obvious  animal  welfare  concerns  are  raised  when  animals  become  overtly  ill  from  an  infection  but  many  infections  of  rodents  do  not  produce  noticeable  sickness.    These  animals  without  clinical  signs  will  remain  undetected  unless  a  strong  colony  surveillance  or  sentinel  program  is  in  place.    When  infected  animals  remain  undetected  and  are  utilized  in  studies,  any  associated  abnormal  physiological  or  behavioral  status  could  lead  to  aberrant  study  results,  sometimes  requiring  a  repeat  of  the  study  with  more  animals.    Once  an  infection  is  detected  in  a  colony,  methods  to  eliminate  or  control  the  spread  of  infection  often  involve  euthanizing  large  numbers  of  animals  in  a  colony  or  facility  as  well.    Therefore,  prevention,  detection,  and  control  of  infections  in  rodent  colonies  supports  the  general  animal  welfare  concepts  of  Reduction,  Replacement,  and  Refinement  by  decreasing  the  likelihood  of  repeating  studies,  preventing  illness,  and  unnecessarily  euthanizing  animals.    Consideration  should  be  given  to  a  routine  auditing  process  by  management  and  others  in  assuring  that  best  biosecurity  practices  are  in  place.    This  will  better  protect  not  only  the  mission  of  the  institution  but  also  the  welfare  of  each  animal  under  its  care.    Fenbendazole  Improves  Pathological  and  Functional  Recovery  Following  Traumatic  Spinal  Cord  Injury.  Chen  Guang  YU,  Ph.D.,  M.D.1,2,  Rajana.  Singh1,2,  Carolyn  A.  Crowdus1,2,  Kashif  Raza1,    Jeanie  F.  Kincer,  D.V.M.,  D.A.C.L.A.M.3,  and  James.  W.  Geddes,  Ph.D.1,2,  1Spinal  Cord  and  Brain  Injury  Research  Center,  2Department  of  Anatomy  and  Neurobiology,  3Division  of  Laboratory  Animal  Resources,  University  of  Kentucky,  Lexington,  Kentucky    During  a  study  of  spinal  cord  injury  (SCI),  mice  in  our  colony  were  treated  with  the  anthelmintic  fenbendazole  to  treat  pinworms  detected  in  other  mice  not  involved  in  our  study.  As  this  was  not  part  of  the  original  experimental  design,  we  subsequently  compared  pathological  and  functional  outcomes  of  SCI  in  female  C57BL/6  mice  who  received  fenbendazole  (150  ppm,  8  mg/kg  body  weight/day)  for  4  weeks  prior  to  moderate  contusive  SCI  (50  kdyn  force)  as  compared  to  mice  on  the  same  diet  without  added  fenbendazole.  The  fenbendazole-­‐treated  mice  exhibited  improved  locomotor  function,    determined  using  the  Basso  mouse  scale,  as  well  as  improved  tissue  sparing  following  contusive  SCI.      Fenbendazole  may  exert  protective  effects  through  multiple  possible  mechanisms,  one  of  which  is  inhibition  of  the  proliferation  of  B  lymphocytes,  thereby  reducing  antibody  responses.  Autoantibodies  produced  following  SCI  contribute  to  the  axon  damage  and  locomotor  deficits.  Fenbendazole  pretreatment  reduced  the  injury-­‐induced  CD45Rpositive  B  cell  signal  intensity  and  IgG  immunoreactivity  at  the  lesion  epicenter  6  weeks  after  contusive  SCI  in  mice,  consistent  with  a  possible  effect  on  the  immune  response  to  the  injury.  Fenbendazole  and  related  benzimadole  antihelmintics  are  FDA  approved,  exhibit  minimal  toxicity,  and  represent  a  novel  group  of  potential  therapeutics  targeting  secondary  mechanisms  following  SCI.    A  Rapidly  Evolving  Shift  in  Monitoring  Schemes  for  Rodent  Pathogens.  Kenneth  S.  Henderson  Ph.D.,  Director,  Research  and  Development,  Research  Animal  Diagnostic  Services,  Charles  River  Laboratories    The  maturation  of  nucleic-­‐acid  based  technologies  and  the  pressure  to  do  more  with  fewer  resources  has  culminated  in  the  development  of  novel  routine  monitoring  schemes  for  laboratory  rodent  pathogens.      The  use  of  soiled  bedding  sentinels  supported  by  traditional  diagnostic  methodologies  has  gone  unchallenged  in  research  vivariums  for  more  than  a  half  a  century.    Publications  have  long  supported  that  many  pathogens  do  not  readily  transfer  to  bedding  sentinels,  but  this  awareness  has  become  augmented  as  the  increase  use  of  PCR  is  providing  data  that  pathogen  transmission  to  bedding  sentinel  is  even  more  problematic  than  realized  a  decade  ago.    The  development  of  evolved  high-­‐throughput  real-­‐time  PCR  assay  panels  provide  a  more  efficient  and  sensitive  alternative  to  traditional  methods.    Implementation  of  PCR-­‐based  testing  of  research  biologics  and  quarantine  has  already  improved  upon  preventing  pathogens  from  entering  into  vivariums.      Now,  PCR  is  successfully  infringing  on  the  traditional  bedding  sentinel  monitoring  of  colony  rodents  by  offering  options  that  reduce  screening  cost  and  resources  as  well  as  support  the  3R  philosophy  by  improving  upon  sentinel  rodent  welfare  or  completely  eliminating  the  use  of  bedding  sentinels.    One  model  of  pathogen  screening  still  continues  the  use  of  sentinels,  but  the  collection  and  simplified  shipment  of  non-­‐invasive  PCR  samples  and  dry  blood  spots  no  longer  requires  live  sentinel  

2014  District  5  AALAS  Meeting  shipment.      A  more  novel  and  recent  scheme  relies  only  on  the  PCR  testing  of  exhaust  air  dust  (EAD)  samples  collected  from  hoses,  plenums  or  filters  and  does  not  require  the  use  of  sentinels.    This  presentation  will  discuss  the  broad  emerging  application  of  PCR  to  rodent  health  monitoring  and  the  associated  advantages  and  disadvantages.      Biology,  Care,  and  Research  Use  of  a  Unique  USDA  Rodent  Species:    The  African  Spiny  Mouse  (Acomys  cahirinus).  Cheryl  L.  Haughton,  D.V.M,  D.A.C.L.A.M.1,  Ashley  W.  Seifert,  Ph.D.2,  1Division  of  Laboratory  Animal  Resources,  2Department  of  Biology,  University  of  Kentucky,  Lexington,  Kentucky    African  spiny  mice  present  some  unique  challenges  in  the  research  environment.    These  USDA  covered  wild  rodents  are  found  throughout  northern  Africa  and  sub-­‐Saharan  Africa.    They  tend  to  live  in  burrows  found  in  arid,  rocky  habitats,  and  they  occasionally  climb.    The  name  “spiny  mice”  refers  to  spiny  hairs  that  cover  most  of  their  dorsal  skin.  Although  pointed,  the  role  of  these  hairs  in  the  wild  is  unknown.    Spiny  mouse  skin  is  incredibly  weak  and  animals  can  lose  pieces  in  response  to  predators  or  during  handling.    African  spiny  mice  are  emerging  as  a  useful  model  for  mammalian  tissue  regeneration  studies  and  as  a  model  for  late  term  pregnancy  defects  during  development.    Spiny  mice  can  regenerate  damaged  tissue  without  scarring.      The  following  are  tips  on  biology,  housing,  handling,  and  care.    Spiny  mice  prefer  warm  temperatures  and  to  live  in  established  social  groups.    Animals  are  predominantly  nocturnal,  although  they  exhibit  bouts  of  activity  throughout  the  day  under  laboratory  conditions.    Preferred  housing  is  a  secure  enclosure  with  solid  walls  or  steel  wire  of  no  greater  than  ½”  x  ½”  size.      Provision  of  items  to  gnaw  help  with  continuous  teeth  growth  and  strong  desire  to  gnaw.    Tubes  or  pipes  to  tunnel  through  are  important  for  cage  enrichment.    Spiny  mice  should  be  acclimated  to  handling.    The  tail  is  devoid  of  hair  and  consists  of  scales.    The  tail  is  fragile  and  can  be  subject  to  de-­‐gloving  injuries  from  inappropriate  capturing  and/or  handling  procedures.    Spiny  mice  are  omnivorous,  but  prefer  to  eat  seeds.    An  adequate  supply  of  food  and  water  is  essential  for  good  health.    Spiny  mice  are  prone  to  obesity,  which  can  result  in  diabetes  mellitus.    Knowledge  of  normal  spiny  mouse  biology  and  behavior  enables  establishment  of  good  care  and  management  practices  to  utilize  spiny  mice  well  in  research.      Leak  Detection  System  (LDS).  Richard  Cluck,  R.L.A.T.G,  Tecniplast  USA,  Exton,  Pennsylvania    Discussion  concerning  the  benefits  of  utilizing  a  leak  detection  system  on  IVC  cage  systems.    Focus  of  the  talk  will  concentrate  on  the  new  Leak  Detection  System  (LDS)  that  Tecniplast  brought  to  the  market  last  year  and  its  success  at  5  beta  sites  and  a  couple  of  demos.  

The  NexGen  Individually  Ventilated  Caging  System.  Marc  Wohlfeil,  Allentown,  Inc.,  Allentown,  New  Jersey    The  NexGen  is  the  new  Individually  Ventilated  Caging  System  offered  by  Allentown,  which  offers  many  improvements  over  previous  IVC  systems  such  as  a  weight  reduction  of  20-­‐25%,  increased  visibility  into  cages,  an  integrated  spring-­‐operated  lock  with  docking  indicator,  improved  ergonomic  handling,  improved  aesthetics,  price  and  delivery  timeline.  The  NexGen  can  be  operated  in  both  positive  and  negative  modes,  while  guaranteeing  exhaust  plenums  are  always  under  negative  pressure,  which  ensures  allergens  and  pathogens  do  not  escape  to  the  room  environment  when  operated  with  empty  stalls  or  when  cages  are  removed.    Managing  The  Macro  Threats  of  Micro  Organisms.  Matthew  Roberts,  E.M.T.,  Quip  Laboratories,  Wilmington,  Delaware    Outbreak  prevention  is  everyone’s  responsibility  so  it’s  important  to  have  more  than  just  a  passing  knowledge  of  biosafety  techniques  and  principles.      The  weakest  link  in  even  the  strongest  chain  can  be  carelessness,  ignorance  or  lack  of  preparedness.    When  it  comes  to  protecting  the  health  of  laboratory  animals,  these  three  factors  can  be  deadly,  costly  and  frustrating.    This  talk  will  describe  some  of  the  ways  in  which  laboratory  animal  professionals  can  enhance  their  role  in  bio-­‐vigilance  by  increasing  their  awareness  of  how  pathogens  make  their  way  into,  and  around,  animal  facilities.    We’ll  also  examine  some  of  the  most  overlooked,  as  well  as  some  of  the  most  common,  roosts  in  which  pathogens  make  their  homes  in  animal  facilities.    Finally,  we’ll  review  the  importance  of  following  facility  hygiene  protocols  and  examine  some  myths  and  misconceptions  about  the  definition  of  “clean,”  “sanitized”  and  “disinfected.”    The  difference  is  important  and  the  level  of  understanding  has  a  real  impact  on  the  health  of  animals  in  our  care,  as  well  as  the  cost  and  quality  of  critical  research  outcomes.      

2014  District  5  AALAS  Meeting  

FRIDAY,  MAY  16  

TRACK  1  PRESENTATION  ABSTRACTS  

Brain  Aging  in  Dogs:    What  can  we  learn  about  Alzheimer’s  disease?  Elizabeth  Head,  M.A.,  Ph.D.,  Sanders-­‐Brown  Center  on  Aging,  University  of  Kentucky,  Lexington,  Kentucky    Aging  dog  naturally  develop  cognitive  decline  in  several  different  domains  (including  learning  and  memory)  but  also  exhibit  human-­‐like  individual  variability  in  the  aging  process.    Cognitive  dysfunction  with  age  may  involve  similar  neuropathology  as  observed  in  the  brains  of  patients  with  Alzheimer’s  disease  (AD).  Specifically,  aged  dogs  show  brain  atrophy,  a  loss  of  neurogenesis  and  growth  factors,  and  an  accumulation  of  oxidative  damage  and  beta-­‐amyloid  pathology.  Cerebrovascular  pathology  also  occurs  in  aging  dogs  and  in  AD.  Thus,  studies  of  treatments  to  improve  cognition  and  reduce  neuropathology  in  aging  dogs  benefits  not  only  the  canine  population  but  also  can  be  translated  to  human  clinical  trials  in  patients  with  AD.      Husbandry,  Veterinary  Support  and  Clinical  Problems  Associated  with  Neurobehavioral  Studies  in  Aged  Canines.  Jeffrey  Smiley,  D.V.M.,  Ph.D.,  D.A.C.L.A.M.,  Division  of  Laboratory  Animal  Resources,  University  of  Kentucky,  Lexington,  Kentucky  

Dr  Smiley  will  give  a  short  presentation  on  husbandry  considerations  and  veterinary  medical  care  and  support  of  aged   beagles   used   in   chronic   neurobehavioral   studies.   Topics   of   discussion  will   include   pre-­‐study   evaluation   of  candidate   animals,   quarantine   procedures,   animal   husbandry   considerations   and   common   medical   problems.      Investigation  of  Cranial  Cruciate  Ligament  Deficiency  and  Associated  Surgical  and  Orthosis  Interventions  Using  Computer   Simulation.   Gina   Bertocci,   Ph.D.,   P.E.,   Department   of   Bioengineering,   University   of   Louisville,   Louisville,  Kentucky    Cranial  cruciate  ligament  (CrCL)  deficiency  affects  the  canine  stifle  (knee)  and  is  one  of  the  most  common  canine  orthopedic  injuries,  having  an  economic  impact  of  more  than  $1  billion  in  the  United  States  in  2003.  CrCL  deficiency  has  a  prevalence  of  2.55%  across  all  breeds  and  is  most  prevalent  in  Newfoundlands  (8.9%),  Rottweilers  (8.3%),  and  Labrador  Retrievers  (5.8%).  Despite  such  high  prevalence,  CrCL  deficiency  is  still  poorly  understood  and  is  thought  to  be  due  to  degradation  and  not  the  result  of  trauma.  Surgical  intervention  is  often  employed  to  stabilize  the  CrCL-­‐deficient  stifle,  but  no  single  surgical  procedure  is  conclusively  supported  to  suggest  long-­‐term  success,  osteoarthritis  prevention  or  superiority.  We  developed  a  canine  pelvic  limb  3D  computer  simulation  model  of  walking  to  gain  an  improved  understanding  of  stifle  biomechanics,  as  well  as  factors  that  may  predispose  dogs  to  CrCL  rupture.  This  model  allows  noninvasive  visualization  and  analysis  of  stifle  biomechanics,  simulates  the  intact  and  CrCL-­‐deficient  stifle,  and  was  utilized  to  investigate  anatomical  characteristics  and  biomechanical  parameters  such  as  ligament  stiffness  and  tibial  plateau  angle  to  gain  an  understanding  of  their  role  in  producing  CrCL  deficiency.    Our  computer  model  was  based  on  a  healthy  male  Golden  Retriever  with  no  orthopedic  or  neurologic  diseases.  A  pelvic  limb  computed  tomography  (CT)  scan  was  conducted  to  obtain  anatomical  geometry,  and  gait  was  recorded  using  a  motion  capture  system  and  force  platform.  The  computer  model  was  developed  in  computer-­‐aided  engineering  software  and  included  canine-­‐specific  bone  geometry,  ligaments,  muscles  and  ground  reaction  forces.  Model  simulation  of  the  stance  phase  of  walking  was  used  to  evaluate  loads  placed  on  stifle  ligaments,  translation  and  rotation  of  the  tibia  relative  to  the  femur,  and  contact  forces  between  the  femur  and  menisci  in  both  the  intact  and  deficient  stifle.    Commonly  employed  surgical  procedures  (tibial  plateau  leveling  osteotomy  (TPLO),  tibial  tuberosity  advancement  (TTA),  lateral  femorotibial  suture  stabilization  (LFTS)  and  TightRopeTM  stabilization  (TR))  were  implemented  in  the  model  through  collaboration  with  a  veterinary  orthopedic  surgeon  to  evaluate  the  ability  of  procedures  to  restore  normal,  CrCL-­‐intact  stifle  biomechanics.  Ligament  loads,  along  with  translation  and  rotation  of  the  tibia  relative  to  the  femur  and  meniscal  loads  were  evaluated  across  surgical  procedures.  Additionally,  we  will  investigate  parameters  specific  to  each  surgical  procedure  (e.g.  suture  tension,  tibial  plateau  rotation  angle,  etc.)  to  further  our  understanding  of  their  influence  on  surgical  efficacy.        Finally,  stifle  orthoses  (braces)  offer  an  alternative  to  surgical  intervention,  especially  in  patients  that  are  poor  anesthesia  candidates  with  significant  co-­‐morbidities,  that  are  of  advanced  age  or  whose  owners  lack  the  financial  means  for  more  

2014  District  5  AALAS  Meeting  costly  surgery.  In  partnership  with  a  veterinary  prosthetics  and  orthotics  clinic  we  are  characterizing  stifle  orthosis-­‐using  dogs.  Additionally,  we  have  implemented  a  a  customized  stifle  orthosis  in  our  computer  model  to  investigate  associated  stifle  biomechanics.  We  have  compared  stifle  biomechanics  during  gait  in  a  CrCL-­‐deficient  stifle  with  and  without  an  orthosis  to  a  CrCL-­‐intact  stifle.  Furthermore,  we  are  in  the  process  of  investigating  stifle  orthosis  design  and  fit  parameters  (e.g.  hinge  type  and  strap  tension),  along  with  patient-­‐specific  parameters,  using  our  computer  model  to  understand  their  role  in  orthosis  effectiveness.    Development  of  a  Bovine  Model  of  Chronic  Ischemic  Heart  Failure  for  MCS  Development  and  Testing.  Leslie  C.  Sherwood,  D.V.M.1,3,  Laura  Lott,  L.A.T.,  S.R.S.1,  ,  Regina  Turner,  A.S.1,3,  Mike  Sobieski,  R.N.,  C.C.P.1,2,  1Cardiovascular  Innovation  Institute,  2Division  of  Thoracic  and  Cardiovascular  Surgery,  3Research  Resources  Facilities,  University  of  Louisville,  Louisville,  Kentucky    Heart  failure  (HF)  is  a  major  and  growing  public  health  concern.    In  2005  approximately  5  million  patients  in  the  United  States  were  suffering  from  heart  failure  at  an  estimated  total  direct  and  indirect  annual  cost  of  27.9  billion  dollars  (AHA,  2005).    Many  of  these  patients  have  reversible  ventricular  dysfunction.    Patients  with  acute  cardiogenic  shock  following  open-­‐heart  surgery  or  patients  with  chronic  heart  failure  due  to  myocardial  ischemia  and/or  infarction  often  do  not  respond  to  pharmacological  therapy  and  require  mechanical  cardiac  support  to  survive.    The  advent  and  widespread  success  of  mechanical  circulatory  support  (MCS)  heralds  a  new  era  of  cardiovascular  medicine  in  which  physicians  and  patients  have  multiple  options  for  the  treatment  of  heart  failure  whether  the  intent  is  to  utilize  the  device  as  a  bridge  to  transplant,  bridge  to  recovery,  or  destination  therapy.    Each  device  entails  unique  surgical  and  physiological  considerations  and  offers  benefits  and  drawbacks  for  the  patient  as  well  as  the  physician.    Device  implantation  strategy,  efficacy,  hemocompatibility,  as  well  as  human  factors  such  as  quality  of  life  during  device  support  and  recovery  after  device  explanation  must  be  considered.  Historically,  MCS  devices  have  been  developed  and  tested  in  animal  models  with  normal  cardiovascular  systems  in  which  device  function  and  hemocompatibility  can  be  assed,  however  with  a  normal  model,  we  are  unable  to  evaluate  the  true  efficacy  of  the  device  as  well  as  further  study  recovery  mechanisms  of  the  heart  while  unloaded  with  a  device.  Until  recently  a  reliable  large  animal  model  of  HF  in  which  to  more  accurately  assess  the  efficacy  of  MCS  devices  was  difficult  to  develop  and  sustain  without  high  mortality.  This  presentation  will  discuss  the  development  of  a  consistent,  reliable  bovine  model  of  chronic  ischemic  heart  failure  via  coronary  microembolization.  Additionally,  a  team  consisting  of  a  human  health  care  provider,  a  veterinarian  and  veterinary  staff  will  cover  anesthetic  induction  and  maintenance,  analgesia,  medical  management  and  husbandry  care.  

       A  special  thank  you  to  our  Silver  Sponsor:  

       

2014  District  5  AALAS  Meeting  

FRIDAY,  MAY  16  

TRACK  2  PRESENTATION  ABSTRACTS    The  Mysteries  of  Microsurgery.  Betsy  F.  Fink,  B.S.,  R.L.A.T.G.,  Division  of  Plastic  and  Reconstructive  Surgery,  University  of  Kentucky,  Lexington,  Kentucky    The   art   and   science   of   microsurgery   will   be   discussed   in   this   presentation,   specifically   as   it   relates   to   the   plastic   and  reconstructive  surgery  and  training  of  our  surgical  residents.  The  technology  of  microscope-­‐directed  surgery  has  presented  a  seemingly   limitless  new  frontier   in  medicine,  bounded  only  by  one’s   imagination.  The  transfer  or  repair  of  nerves,  vessels,  composite  tissue  parts;  the  study  of  growth,  healing,  transplantation,  fertility,  physiology;  the  response  of  microsutures  to  trauma  or  pharmacologic  agents;  the  innovative  use  of  prosthetic  material;  and  laser  instrumentation  are  merely  a  portion  of   the  spectrum  encompassed  by  microsurgery.    A  prerequisite   to   investigative  or  clinical  microsurgery   is  experience  with  the  use  of  the  microscope  and  microsurgical  instrumentation.    In  order  to  perform  such  tedious  and  intricate  procedures  one  must  be  adequately   trained.  The  purpose  of  our   training  course   is   to  provide  a  core   framework   for   learning  microsurgical  technique,  and  serves  as  the  open  door  for  one’s  microsurgical  explorations.  Each  stage  of  the  course  is  mastered  through  a  multifaceted   approach;   a   reading   assignment,   an   audiovisual   taped   lecture,   and   an   animal   practicum,   directed   by   the  instructor.    The  student,  typically  surgical  residents,  is  also  educated  in  the  process  of  IACUC  and  federal  regulations  relating  to  use  of  animals  for  research  and  training  to  gain  appreciation  for  the  privilege  of  using  animals  to  benefit  their  expertise.    The  Intersection  of  Public  Health,  Human  Behavior  and  Animals.  John  Poe,  D.V.M.,  M.P.H.,  State  Public  Health  Veterinarian,  Kentucky  Department  for  Public  Health,  Frankfort,  Kentucky    Will  discuss  the  daily  real  challenges  of  a  state  health  dept.  consultative  service  in  the  state  of  Kentucky.  Dr.  Poe  has  spent  over  20  years  practicing  veterinary  medicine  in  Kentucky.  In  addition  to  his  private  practice  he  served  as  a  Veterinary  Medical  Officer  for  the  Animal  and  Plant  Health  Inspection  Service  (APHIS)  branch  of  the  United  States  Department  of  Agriculture  (USDA)  for  five  years.  Dr.  Poe  became  the  Kentucky  State  Public  Health  Veterinarian  five  years  ago  for  the  Kentucky  Department  for  Public  Health.  His  current  responsibilities  include  maintaining  up-­‐to-­‐date  scientific  knowledge  and  training  in  order  to  serve  as  the  department’s  content  expert  for  control  of  zoonoses,  tick-­‐borne  diseases,  arboviruses  and  other  communicable  diseases.  He  provides  consultations  to  health  professionals  and  the  public  regarding  communicable  diseases,  animal  bites  and  zoonoses.  He  provides  educational  training  to  the  staff  and  the  public,  as  well  as  reviewing  legislation  pertinent  to  areas  of  expertise  and  drafts  legislation  and  regulations  to  protect  the  public.  

Taking  Your  Career  Outside  of  the  Box.  Verda  Antoinette  Davis,  B.S.,  M.B.A.,  R.L.A.T.G.,  Department  of  Anatomy  and  Neurobiology,  University  of  Kentucky,  Lexington,  Kentucky    Have  you  ever  wondered  what  other  opportunities  there  are  for  you  to  be  involved  in  laboratory  animal  science?    Maybe  you  are  interested  in  higher  education,  but  don’t  know  what  kinds  of  programs  are  out  there.    Maybe  you  don’t  know  about  the  professional  development  opportunities  available  from  National  AALAS  or  your  local  branches.    Maybe  you  could  earn  AALAS  certifications  or  attend  ILAM  training.    Maybe  you  could  join  LAMA  or  work  towards  SRS  certification.    Don’t  know  what  these  acronyms  mean?    Come  to  this  session  and  find  out  about  opportunities  to  take  your  lab  animal  career  outside  the  box  and  into  a  whole  new  direction!      

Assessing  Animal  Welfare  in  a  Zoological  Setting.  Roy  Burns  D.V.M.,  Louisville  Zoological  Garden,  Louisville,  Kentucky    Defining,  evaluating,  and  improving  animal  welfare  in  zoos  is  an  active  area  of  research  worldwide.    Regulatory  agencies,  zoos,  the  general  public,  and  special  interest  groups  participate  in  discussion  of  zoo  animal  welfare  in  scientific  literature  and  in  popular  media.    Defining  animal  welfare  has  evolved  from  a  focus  on  avoidance  of  a  negative  state  of  welfare  to  a  focus  on  providing  positive  opportunities  that  improve  welfare.      Evaluation  of  animal  welfare  has  grown  from  measuring  resources  provided  to  a  group  of  animals  to  evaluation  of  an  individual  animal’s  behavior,  personality  and  emotional  state.    Biological  markers  (e.g.  cortisol  metabolites)  are  also  used  to  evaluate  animal  welfare.    Enrichment  is  a  common  tool  used  to  improve  the  well-­‐being  of  animals  in  zoos.    Enrichment  practices  provide  opportunities  to  increase  feeding  behaviors  and  provide  sensory  and  social  stimuli.    Cognitive  stimuli,  training,  providing  choice  and  control  over  environment,  and  keeper-­‐

2014  District  5  AALAS  Meeting  animal  relationships    have  recently  been  added  to  the  tool  box  for  improving  animal  well-­‐being  in  zoos.    Louisville  Zoological  Garden  has  been  a  leader  in  new  efforts  in  evaluating  and  improving  the  well-­‐being  of  certain  species  such  as  polar  bears,  elephants,  and  apes.    POSTER  ABSTRACTS  

1. Evaluation  of  Carbon  Dioxide  Dissipation  within  a  Euthanasia  Chamber.  Shelly  M.  Djoufack-­‐Momo1,  Ashlee  Amparan1,  Beverly  Grunden2  and  *Gregory  P.  Boivin3,4,5,  1Department  of  Community  Health  2Department  of  Mathematics  and  Statistics  3Department  of  Pathology  4Department  of  Orthopaedic  Surgery,  Wright  State  University,  Dayton,  OH  5Veterans  Affairs  Medical  Center,  Cincinnati,  Ohio    Carbon  dioxide  (CO2)  is  widely  used  for  small  laboratory  animals  such  as  rodents  for  euthanasia.  The  American  Veterinary  Medical  Association  (AVMA)  guidelines  state  that  “The  practice  of  immersion,  where  conscious  animals  are  placed  directly  into  a  container  prefilled  with  100%  CO2,  is  unacceptable”.  A  common  necessity  in  many  research  facilities  is  to  euthanize  mice  in  sequential  batches.    The  aim  of  this  study  was  to  identify  how  long  it  takes,  after  one  group  of  mice  had  been  euthanized,  for  dissipation  of  CO2  within  a  euthanizing  chamber  before  a  second  animal  or  group  of  animals  could  be  put  in  the  chamber.  Several  parameters  were  examined  to  measure  the  influence  on  the  CO2  dissipation  rate.  These  included  flow  rate,  chamber  position,  presence  and  removal  of  a  cage  from  the  chamber,  and  whether  the  lid  was  left  on  or  removed.  CO2  peaked  between  50-­‐80%  in  the  chamber  for  all  trials.  The  concentration  of  CO2  in  all  trials  dropped  within  2  minutes  to  below  10%  CO2  except  when  the  lid  was  left  on  the  chamber  (dissipation  took  over  20  minutes).  CO2  dissipation  was  significantly  faster  when  the  chamber  was  turned  upside  down  following  filling  (p<0.0001).  Only  leaving  the  lid  on  the  chamber  had  any  practical  implications  for  delaying  CO2  dissipation.  We  recommend  that  2  minutes  be  allowed  for  CO2  to  clear  from  the  chamber  prior  to  subsequent  euthanasia  procedures  unless  the  chamber  is  manipulated  to  increase  the  dissipation  rate.    

2. Carbon  dioxide  exposure  of  research  personnel  during  euthanasia  procedures.  Ashlee  A.  Amparan1,  Shelly  M.  Djoufack-­‐Momo  

1,  Beverly  Grunden2,  Gregory  P.  Boivin3,4,5,  1Department  of  Community  Health  2  Department  of  Mathematics  and  Statistics  3  Department  of  Pathology  4  Department  of  Orthopaedic  Surgery,  Wright  State  University,  Dayton,  OH  5  Veterans  Affairs  Medical  Center,  Cincinnati,  Ohio    Carbon  dioxide  (CO2)  is  one  of  the  most  commonly  used  euthanasia  agents  in  laboratory  animal  facilities.  Euthanasia  is  usually  performed  from  a  compressed  gas  cylinder  supplying  CO2  to  a  small  chamber.  Considerable  research  has  gone  into  the  effect  of  the  agent  on  animals,  but  little  has  been  done  to  examine  the  potential  effects  on  laboratory  personnel  that  are  euthanizing  animals.  The  Occupational  Safety  and  Health  Administration  (OSHA)1  set  threshold  limit  values  (TLV)  that  all  employers  are  required  to  abide  by  in  order  to  protect  workers  exposed  to  CO2.  The  time-­‐weighted  average  (TWA)  is  5,000  parts  per  million  (ppm).  The  short-­‐term  exposure  limit  (STEL)  is  30,000  ppm.  This  study  examined  CO2  exposure  during  CO2  euthanasia  procedures.  We  hypothesized  that  the  CO2  levels  will  meet  OSHA  standards  and  that  CO2  concentrations  will  depend  on  flow  rate,  distance  from  the  chamber,  and  room  ventilation.  Under  all  conditions,  CO2  concentrations  increased  significantly  in  the  room  to  between  600-­‐4000  ppm.  The  CO2  levels  remained  below  the  Occupational  Safety  and  Health  Administration’s  threshold  limit  values.  The  results  show  that  under  several  different  modeling  conditions  there  is  no  occupational  threat  of  CO2  exposure  during  routine  rodent  euthanasia  procedures.    

3. Genotyping  DNA  isolated  using  cross-­‐linked  iminodiacetate  styrene  divinylbenzene  copolymer  beads  (Chelex).  Boivin  GP,  Otoñio-­‐Rivera  V,  Boakye  A,  Grobe  N,  Di  Fulvio  M.,  Boonschoft  School  of  Medicine,  Wright  State  University,  Dayton  Ohio      Genotyping  mice  rely  on  isolation  of  DNA  from  tissues  typically  involving  painful  procedures  such  as  tail  snipping,  digit  removal  or  ear  punch  and  the  use  of  expensive  kits.  Although  harvesting  of  hair  has  been  proposed  in  the  past  as  a  source  for  genomic  DNA,  there  has  been  a  perceived  complication  because  of  low  DNA  yields  and  fear  of  contamination.  We  hypothesized  that  DNA  isolation  from  hair  follicles  will  be  as  effective  as  more  painful  procedures  for  mice  genotyping.  We  tested  an  adapted,  simplified,  fast  and  cheap  version  of  a  common  forensic  method  used  for  fingerprinting  human  samples  and  compared  the  results  against  tissue-­‐based  enzymatic  commercial  kits.  As  source  of  DNA  for  genotyping,  we  used  10-­‐20  hair  follicles  plucked  from  3  different  groups  of  genetically  engineered  mice.  Briefly,  over  200  samples  of  hair  follicles  with  appropriate  positive  and  negative  controls  were  directly  placed  in  10%  Chelex  resin  and  incubated  20  min  at  100ºC  with  gentle  swirling.  An  aliquot  of  the  supernatant  corresponding  to  1-­‐2%  of  the  total  volume  containing  follicle's  DNA  released  by  the  action  of  the  resin  was  directly  used  as  template  for  hot-­‐start  polymerase  chain  reactions  (PCR)  coupled  to  custom-­‐designed  gene-­‐specific  oligonucleotide  primers.  The  results  demonstrate  that  our  procedure  provides  specific  and  accurate  genotyping  results  in  less  than  4h  at  a  cost  per  PCR  of  a  tenth  of  commercially  available  kits.  We  recommend  that  in  circumstances  where  haired  mice  can  be  analyzed,  the  use  of  Chelex-­‐digested  hair  follicles  is  an  excellent  source  of  DNA  for  PCR  genotyping  of  mice.  

2014  District  5  AALAS  Meeting  4. Non-­‐surgical  Embryo  Transfer  In  Mice  Is  An  Easy,  Effective,  and  Ethical  Replacement  For  Surgery.  Kendra  

Steele,  Barbara  Stone,  James  Hester,  Angelika  Fath-­‐Goodin,  ParaTechs  Corporation,  Lexington  Kentucky    Surgical  embryo  transfer  (ET)  is  an  effective  method  to  deposit  embryos  into  the  uterine  horn  of  mice.    However,  surgery  is  expensive,  time-­‐consuming,  and  requires  technical  expertise.    Surgery  is  also  a  stressful  procedure  for  the  mouse,  which  has  to  be  anesthetized  and  treated  with  an  analgesic.  We  have  developed  a  simple,  brief  procedure  for  ET  using  a  non-­‐surgical  device,  and  our  hypothesis  is  that  this  non-­‐surgical  procedure  is  less  stressful  for  the  mouse,  as  effective  as  surgical  ET,  and  the  procedure  can  be  repeated  on  the  same  mouse.    In  order  to  compare  the  effectiveness  between  the  non-­‐surgical  and  surgical  procedures,  we  performed  side-­‐by-­‐side  comparisons  with  20  mice  per  method  and  repeated  the  experiment  using  four  different  strains.    Pregnancy  rate  is  higher  in  mice  that  have  undergone  non-­‐surgical  ET  than  mice  subjected  to  surgery,  and  litter  size  and  birth  rate  from  the  two  procedures  are  similar.  We  then  performed  the  non-­‐surgical  method  up  to  two  more  times  on  individual  CD-­‐1  mice,  each  time  allowing  the  mouse  to  recover  for  at  least  20  days  post-­‐partum  before  becoming  pseudopregnant  again.    The  data  demonstrate  that  the  non-­‐surgical  ET  procedure  can  be  used  multiple  times  on  a  mouse,  but  there  is  a  reduced  pregnancy  rate.  Since  non-­‐surgical  ET  does  not  require  sedation,  opening  of  the  inner  body  cavity,  or  use  of  an  analgesic,  we  hypothesized  that  this  procedure  is  less  stressful  for  the  mouse  than  surgery.    We  used  electrocardiography  (n=11)  and  fecal  corticosterone  ELISA  (n=15)  to  monitor  pseudopregnant  mice  that  underwent  anesthesia  only,  non-­‐surgical  ET  with  and  without  anesthesia,  or  surgery.  Our  results  show  that  the  non-­‐surgical  procedure  without  anesthesia  does  not  affect  heart  rate  or  alter  the  levels  of  the  stress  biomarker  fecal  corticosterone,  whereas  surgery  and  anesthesia  alone  lower  heart  rate  for  at  least  one  hour  after  administration  and  increase  levels  of  fecal  corticosterone.  Responsible  animal  scientists  are  required  to  follow  Russell  and  Burch’s  “3Rs”  of  animal  research:    to  replace,  reduce,  and  refine.    The  non-­‐surgical  procedure  refines  the  ET  method  by  minimizing  animal  stress,  and  can  reduce  the  number  of  mice  needed  for  experiments,  offering  an  advantageous  alternative  to  surgical  transfer.    

5. A  Non-­‐surgical  Uterine  Transfer  Technique  for  Mouse  Embryos  after  Cryopreservation,  In  Vitro  Fertilization,  ES-­‐cell  Injection,  and  Sperm  during  Artificial  Insemination.  Barbara  Stone,  ParaTechs  Corporation,  Lexington  Kentucky    Many  procedures  used  for  assisted  reproductive  techniques  can  be  damaging  to  the  integrity  of  embryos.    Therefore,  we  chose  to  determine  if  embryos  could  be  successfully  transferred  with  non-­‐surgical  methods  after  cryopreservation,  in  vitro  fertilization  (IVF),  or  embryonic  stem  (ES)  cell  injection  as  an  alternative  to  traditional  surgical  embryo  transfer.    The  non-­‐surgical  embryo  transfer  or  NSET  technique  requires  the  use  of  a  device  that  has  a  tapered  Teflon  catheter  capable  of  precise  liquid  delivery.    Once  embryos  are  loaded  into  the  device,  the  catheter  passes  through  the  vagina  and  traverses  the  cervix  to  deposit  embryos  into  the  uterine  horn  of  a  recipient  mouse.  The  NSET  technique  was  used  to  transfer  cryopreserved  B6C3F2  1-­‐cell  embryos  cultured  to  blastocysts  to  female  CD-­‐1  recipients.    These  transfers  resulted  in  a  birth  rate  of  39.4%  (N=16).  IVF  of  fresh  oocytes  from  B6C3F1  female  mice  and  cryopreserved  sperm  from  B6C3F1  males  was  performed.    Fertilized  embryos  were  cultured  to  blastocyst  stage  and  transferred  to  CD-­‐1  recipients.    Transfer  of  IVF-­‐derived  blastocysts  resulted  in  a  birth  rate  of  43.3%  (N=20).    C57BL/6  blastocysts  were  injected  with  JM8A3.N1  ES  cells  and  transferred  to  CByB6F1/J  recipients.    The  birth  rate  from  NSET  transfers  of  ES  cell-­‐injected  blastocysts  was  30.5%  (N=25).    These  results  indicate  that  the  non-­‐surgical  transfer  of  blastocysts  is  effective  for  transfer  after  embryo  manipulations  and  cell  culture.      We  also  hypothesized  that  the  non-­‐surgical  transfer  technique  could  be  used  for  delivery  of  sperm  for  artificial  insemination.    Fresh  sperm  from  B6C3F1  males  was  transferred  to  the  uterine  horn  of  hormone-­‐induced  CD-­‐1  females,  resulting  in  live  births  and  a  36%  pregnancy  rate  (N=19).    The  non-­‐surgical  transfer  technique  for  embryos  or  sperm  is  fast,  does  not  require  anesthesia  or  analgesia,  and  post-­‐procedure  recovery  is  not  necessary.    These  results  provide  proof  that  the  technique  successfully  produces  live  pups  after  transfer  of  1)  embryos  after  cryopreservation,  IVF,  or  ES-­‐cell  injection,  and  2)  sperm  during  artificial  insemination.    

6. Targeted  Delivery  of  Magnevist®  to  the  Midbrain:  A  Proof-­‐of-­‐concept  Study  for  Parkinson’s  Disease  Treatment.  April  Evans1,  Ryan  Weeks1  Peter  A.  Hardy1,  Eric  S.  Forman1,  Francois  Pomerleau1,  Jorge  E.  Quintero1,  Greg  A.  Gerhardt1,  Don  M.  Gash1,  David  A.  Bumcrot2,  Zhiming  Zhang1,  and  Richard  Grondin1,  1University  of  Kentucky  Medical  Center,  Department  of  Anatomy  &  Neurobiology,  Lexington,  Kentucky,  2Editas  Medicine,  Cambridge,  MA  02142;  previously  at  Alnylam  Pharmaceuticals,  Inc.,  Cambridge,  Massachusetts      Direct  drug  administration  into  the  brain  parenchyma  offers  a  high  degree  of  targeting,  while  reducing  the  risks  of  unwanted  side-­‐effects  by  limiting  systemic  exposure.  However,  targeted  drug  delivery  to  the  midbrain  region  remains  a  challenge.  Here,  we  used  an  investigational  needle-­‐tip  catheter  and  cranial  anchor  system  coupled  to  implanted  programmable  pumps  to  continuously  infuse  5mM  gadopentetate  dimeglumine  (Magnevist®)  into  the  substantia  nigra  (SN)  of  female  rhesus  monkeys  to  lay  the  foundation  for  delivering  drug  therapies  to  the  midbrain  region.  Flow  rate  tolerability  of  either  0.1  µL/min  (144  µL/day)  to  the  right  SN  for  7  days  or  0.2  µL/min  (288  µL/day)  to  the  left  SN  for  7  days  was  assessed  by  monitoring  clinical  observations,  body  weights  and  food  consumption  measurements.  Evaluation  of  post-­‐surgical  MRI  indicated  that  the  placement  of  each  intranigral  catheter  was  within  a  2-­‐mm  radius  from  the  intended  surgical  target  and  that  all  catheters  were  patent,  as  evidenced  

2014  District  5  AALAS  Meeting  by  the  presence  of  Magnevist®  at  the  catheter  tip.  In  addition,  post-­‐surgical  MRI  evaluation  indicated  that  the  volume  of  distribution  achieved  in  the  midbrain  region  with  Magnevist®  infused  at  a  rate  of  0.2  μL/min  was  greater  than  that  achieved  at  0.1  μL/min  by  nearly  2-­‐fold.  There  were  no  indications  of  toxicity  as  noted  in  clinical  observations.  Also,  there  was  no  difference  between  the  two  infusion  rates  with  respect  to  changes  in  body  weight  values  and  no  indication  of  adverse  effects  from  either  Magnevist®  flow  rates  on  appetite  (food  consumption).  Our  data  support  that  direct  drug  delivery  to  the  midbrain  maybe  used  for  the  treatment  of  Parkinson’s  disease.  Based  on  the  data  reported  here,  ongoing  studies  in  non-­‐human  primates  are  evaluating  the  targeted  delivery  to  the  midbrain  of  an  siRNA  designed  to  suppress  alpha-­‐synuclein  expression  as  a  potential  therapy  for  Parkinson’s  disease.  Supported  by:  Alnylam  Pharmaceuticals  Inc.,  and  the  Michael  J.  Fox  Foundation  for  Parkinson’s  Research.  Medtronic  Inc.  provided  the  pumps,  catheters  and  cranial  anchor  system.    

7. Pharmacokinetics  and  Excreta  Recovery  of  [14C]Erioglaucine  to  Determine  the  Impact  of  Solid-­‐Bottom  Versus  Wire-­‐Bottom  Caging  in  Sprague-­‐Dawley  Rats.  Nicole  Vilminot,  MPI  Research,  Mattawan,  Michigan    The  Guide  For  the  Care  and  Use  of  Laboratory  Animals  recommends  the  use  of  solid-­‐bottom  caging  rather  than  wire-­‐bottom  caging  in  rodents  for  many  reasons,  including  reduction  in  environmental  stress,  allowance  for  species-­‐typical  behaviors,  and  decreasing  the  incidence  of  pododermatitis  and  other  types  of  foot  lesions.    In  toxicological  facilities,  historically,  there  has  been  a  concern  for  re-­‐exposure  to  test  article  due  to  coprophagia  when  rodents  are  housed  in  solid-­‐bottom  caging,  which  is  used  as  a  scientific  justification  for  the  use  of  wire-­‐bottom  caging.    Our  Institutional  Animal  Care  and  Use  Committee  (IACUC)  questioned  whether  housing  rats  in  solid-­‐bottom  caging  rather  than  wire-­‐bottom  caging  would  interfere  with  data  interpretation  on  toxicological  studies.    Coprophagia  is  a  normal  behavior  in  rodents  that  occurs  as  the  fecal  pellet  is  being  expelled  from  the  anus  rather  than  while  lying  on  cage  bottom  floor.    Nutritional  studies  that  require  complete  elimination  of  coprophagia  must  use  special  collars  or  fecal  cups,  as  wire  bottom  caging  is  not  sufficient  to  control  the  behavior.    Furthermore,  between  10-­‐50%  of  feces  is  re-­‐ingested,  so  even  with  a  test  article  that  is  excreted  100%  in  active  form  in  the  feces,  at  most,  this  would  only  increase  exposure  by  50%.    Different  dose  groups  in  toxicological  studies  are  often  dosed  at  5  to  10-­‐fold  dosing  intervals,  which  would  likely  obscure  any  variability  introduced  by  re-­‐ingesting  of  test  article  in  the  feces  (Neale,  1984).    To  determine  the  extent  to  which  animals  housed  in  solid-­‐bottom  caging  are  exposed  to  re-­‐ingestion  of  test  article  compared  to  animals  housed  in  wire-­‐bottom  cages,  two  groups  of  rats  were  housed  individually  in  either  wire-­‐bottom  or  solid-­‐bottom,  bedded  cages.    All  animals  were  administered  a  single  dose  of  [14C]Erioglaucine  (FD&C  Blue  Dye  #1)  via  oral  gavage.    This  compound  was  chosen  because  it  is  largely  excreted  in  the  feces  unchanged  within  36  hours  of  administration.    Post-­‐dose,  serial  plasma  samples  and  feces  samples  were  collected  for  radioanalysis.    After  the  final  excreta  sample  collection  at  72  hours,  all  animals  were  euthanized  and  the  residual  carcasses  were  solubilized  and  analyzed  for  total  radioactivity.    Results  showed  no  appreciable  difference  in  either  the  plasma  levels  or  total  carcass  recovery  of  the  [14C]Erioglaucine  between  the  two  groups,  indicating  that  there  was  no  net  increase  in  plasma  exposure,  or  in  the  amount  of  test  article  retained  in  the  gastrointestinal  tract  from  re-­‐ingested  feces  for  rats  housed  in  solid-­‐bottom,  bedded  cages  versus  those  housed  in  wire-­‐bottom  cages.    

8. Comparison  of  Two  Variations  of  Blood  Collection  for  Sentinel  Surveillance  Testing  in  Mice.  Ronda  Combs,  B.S.,  R.L.A.T.,  Glenn  Florence,  L.A.T.G.,  and  Jeanie  F.  Kincer,  D.V.M.,  D.A.C.L.A.M.,  Division  of  Laboratory  Animal  Resources,  University  of  Kentucky,  Lexington,  Kentucky    The  rodent  sentinel  program  is  an  essential  component  in  laboratory  animal  research.  It  continues  to  be  the  primary  means  of  detecting  specific  infectious  pathogens  within  facilities.  In  this  study,  our  goal  was  to  compare  two  variations  of  sentinel  testing  to  see  if  we  could  reduce  animal  numbers,  costs,  and  time.    Female  CD-­‐1  sentinels  were  set  up  in  two  groups  on  18  racks  (9  racks  /  group).  Group  1  was  setup  in  our  standard  housing  arrangement  of  3  mice  per  IVC  cage,  one  cage  per  rack.  At  the  time  of  collection  one  sentinel  was  removed  from  each  cage  for  terminal  sample  collection.  The  sentinel  was  euthanized  and  blood  collected  via  cardiac  puncture.  Evaluation  for  ectoparasites  by  fur  pluck  testing  and  pelt  examination,  gross  necropsy  for  lesioned  organs  and  examination  for  endoparasites  by  anal  tape  test  of  the  perineum  and  visualization  of  cecal  contents  were  performed.  Blood  samples  were  spun  and  serum  submitted  to  a  commercial  laboratory  for  testing.  Group  2  was  setup  with  the  housing  arrangement  of  2  sentinels  per  IVC  cage,  one  cage  per  rack.  At  the  time  of  collection,  one  sentinel  was  selected  from  each  cage  and  placed  back  in  the  home  cage  after  sample  collection.  Blood  was  collected  via  saphenous  vein  and  placed  on  a  blood  spot  serologic  assay  provided  by  a  commercial  diagnostic  laboratory.  Evaluation  for  ectoparasites  by  fur  pluck  testing  and  endoparasites  by  anal  tape  test  of  the  perineum  from  each  sentinel  were  also  performed.  Predominantly,  we  found  group  2  to  be  an  effective  means  of  sentinel  testing  that  reduces  cost,  time  and  most  importantly  reduces  the  number  of  sentinel  animals  required.      

2014  District  5  AALAS  Meeting    

9. Development  and  Implementation  of  an  Ulcerative  Dermatitis  Scoring  System  and  Treatment  Plan  in  Laboratory  Mice.  Leslie  Pittsley,  L.V.T.,  L.A.T.,  Natalie  Madej,  L.A.T.,  and  Kathleen  Patterson,  D.V.M.,  D.A.C.L.A.M.,  MPI  Research,  Mattawan,  Michigan    Ulcerative  dermatitis  is  a  common  clinical  condition  seen  in  laboratory  mice.    A  single  cause  has  not  been  identified  and  the  condition  is  thought  to  be  the  result  of  a  combination  of  genetic,  environmental,  and  pathogenic  components.  Ulcerative  dermatitis  may  occur  spontaneously  or  secondary  to  an  excoriation.  These  lesions  can  allow  for  bacterial  proliferation  of  the  affected  area  and  progression  of  the  condition.    Early  signs  of  ulcerative  dermatitis  include  alopecia,  erythema,  pruritus,  and  scabbing.  Ulcerative  dermatitis  can  progress  rapidly  from  a  simple  excoriation  of  the  skin  to  complete  ulceration  of  the  area  with  loss  of  the  epidermis  and  dermis.    Progression  can  occur  rapidly;  therefore  early  intervention  is  a  key  component  to  successful  treatment.    At  our  facility,  we  developed  and  implemented  an  ulcerative  dermatitis  scoring  system  that  classifies  lesions  into  one  of  three  categories  and  assigns  a  severity  score.  Our  system  uses  three  categories  ranging  from  mild  (score  of  1)  to  severe  (score  of  3).    Each  score  has  an  associated  description  of  clinical  observations  in  order  to  help  technicians  determine  which  score  to  apply  to  an  animal.  Using  the  score  and  considering  the  animal’s  overall  condition  and  study  goals,  a  treatment  plan  is  outlined.    At  a  minimum,  each  plan  includes  toe  nail  trimming  to  reduce  mechanical  trauma  and  additional  environmental  enrichment  to  promote  species  specific  behaviors.    Depending  on  the  score  assigned  to  the  animal  and  study  goals,  topical  treatments  may  be  utilized.    Implementation  of  this  scoring  system  has  allowed  our  team  to  identify  and  treat  ulcerative  dermatitis  cases  quickly  and  consistently  throughout  our  facility.      

10. Performance  of  Three  Types  of  Rodent  Bedding  in  Static  and  Ventilated  Caging  Systems.  L.  Steiner,  E.  Vernasco-­‐Price,  C.  Maute,  R.  Dysko,  Unit  for  Laboratory  Medicine  Ann  Arbor,  Michigan    A  previous  study  at  our  institution  evaluated  three  types  of  bedding  in  a  polyuric  mouse  colony,  assessing  cage  changing  frequency  as  well  as  labor  and  bedding  costs.    A  more  detailed  study  with  the  same  3  bedding  types  was  performed,  comparing  cages  of  polyuric  mice  to  mice  with  normal  urination  rates,  this  time  also  determining  ammonia  levels  as  well  as  comparing  static  cages,  cages  receiving  house  ventilation,  and  cages  receiving  blower  pack  ventilation.    Five  cages,  ranging  in  housing  density  from  1  to  5  mice,  were  assigned  to  one  of  18  groups,  and  were  evaluated  over  a  2  week  period.    Bedding  1  (B1)  was  combination  corncob  bedding,  bedding  2  (B2)  consisted  of  compressed  cellulose,  and  bedding  3  (B3)  was  pelletized  corncob  that  swells  and  breaks  apart  when  it  becomes  saturated.    For  each  static  cage,  300  ml  of  B1  and  B2  and  250  ml  of  B3  were  added  to  the  cages.    For  each  ventilated  cage,  225  ml  of  each  type  of  bedding  was  used.    Cage  dimensions  were  7.5  x  11.75  x  5  inches.    A  colorimetric  ammonia  sensor  was  suspended  in  one  cage  in  each  of  the  18  groups,  and  sensors  were  moved  between  cages  and  groups  upon  cage  change.    Each  cage  was  observed  daily  for  bedding  saturation  levels  and  the  color  of  the  ammonia  sensor  was  noted.    The  cage  was  changed  if  at  least  ¼  of  the  bedding  appeared  wet  or  if  the  ammonia  sensor  registered  more  than  25  ppm.    There  was  no  difference  in  the  number  of  cage  changes  for  normal  urinating  mice  for  all  3  bedding  and  housing  types.    For  polyuric  mice,  there  were  58  additional  cage  changes  for  B1  during  the  2  weeks,  67  additional  changes  for  B2,  and  52  additional  changes  for  B3.    Regarding  ammonia  levels,  B1  had  6  readings  over  25  ppm;  B2  had  18  readings  over  25  ppm;  B3  had  2  readings  over  25  ppm.    Based  on  our  data  for  all  cage  ventilation  and  bedding  types,  the  house  ventilation  system  performed  better  than  static  housing  or  blower  packs.    Overall,  B3  performed  best  regarding  ammonia  levels  and  cage  changing  frequency.    However,  the  dust  created  by  B3  posed  an  increased  risk  of  blocking  the  ventilation  port  (located  at  the  cage  bottom)  and  interfering  with  the  water  source  (either  sipper  tube  or  watering  valve),  causing  cages  to  become  saturated;  this  development  should  be  taken  into  consideration  when  selecting  a  bedding  type.    

11. Farrowing  Process  of  Newborn  Piglets.  Farthing  J,  McKown  G,  Hobson  D,  Hickman  DL,  Laboratory  Animal  Resource  Center,  Indiana  University,  Indianapolis,  Indiana    Farrowing  piglets  is  a  time  consuming  yet  rewarding  process.    Proper  handling  of  newborn  piglets  is  important  for  the  overall  health  and  success  of  a  colony.    Making  sure  they  receive  critical  care  the  first  few  hours  after  birth  helps  achieve  this  goal.    Having  procedures  in  place  and  the  necessary  supplies  gathered  together  before  farrowing  relieves  stress  for  both  the  technicians  and  the  animals.    For  our  purposes,  we  placed  all  the  supplies  we  would  need  into  a  dedicated  bag  used  just  for  farrowing.    It  includes  supplies  such  as  antibiotics,  vitamins,  and  other  necessary  drugs  during  the  process.    It  also  has  scissors,  umbilical  wrap,  iodine,  a  thermometer,  and  gauze.    Also  in  the  room  is  an  ample  supply  of  towels  and  blankets  and  important  phone  numbers  displayed  on  the  wall.    By  pulling  all  these  supplies  together  in  one  place,  it  allows  the  technician  to  relax  and  focus  on  the  farrowing  process.    A  special  thank  you  to  Dr.  Jose  Estrada  for  allowing  us  to  photograph  his  animals.        

2014  District  5  AALAS  Meeting    

12. “Happy  Sheep”:  Human  Interaction  with  Sheep  used  in  Long-­‐Term  Research.  Angelynn  Perchermeier  and  Marissa  Dapore,  Cincinnati  Children’s  Hospital,  Cincinnati,  Ohio    At  Cincinnati  Children’s  Research  Foundation  we  have  a  large  colony  of  long-­‐term  sheep.    Sheep,  especially  those  used  in  long-­‐term  research,  require  daily  human  interaction  in  order  to  provide  an  environment  where  anxiety  is  decreased,  thus  creating  a  safer  environment  and  overall  better  research  models.      By  working  with  and  training  our  sheep  for  haltering  and  crating,  we  have  seen  a  great  improvement  in  undesirable  stress  behaviors  that  tend  to  be  common  during  crating  procedures  for  surgery  and  haltering  for  shearing,  hoof  trims,  etc.    Hand  feeding  enrichment  has  also  helped  us  achieve  this  goal(s);  It  decreases  the  possible  dangers  of  working  with  large  animals  for  daily  husbandry  procedures  such  as  cleaning  and  feeding.  Advantages  from  daily  human  interactions  outweigh  any  disadvantages  that  we  have  encountered  such  as  unwanted  behavior  for  attention  or  an  apparent  developed  need  for  interaction.  We  have  discovered  that  positive  daily  human  interaction  leads  to  what  we  consider  to  be  “happy”,  healthier  sheep  and  has  lessened  the  likelihood  of  injury  to  caretakers.      

13. Beneficial  Forms  of  Enrichment  for  Felines.  Rhea  Waltz,  Cincinnati  Children’s  Hospital,  Cincinnati,  Ohio    All  species  can  greatly  benefit  from  enrichment,  both  environmental  and  social.    At  Cincinnati  Childrens  Research  Foundation  we  have  an  intermittent  short-­‐term  colony  of  felines  that            eventually  are  adopted  out  or  donated  to  zoos  for  breeding  research.      We  are  able  to  provide  novel  enrichment  strategies  through  the  housing  environment,  human  interaction,  socialization,  and  food  treats.      Our  environmental  enrichment  for  felines  consists  of  toys,  furniture,  and  objects,  while  maintaining  a  clean  and  safe  environment.  Cats  are  very  social  creatures  thus  making  it  very  important  to  stimulate  them  and  provide  social  interactions  with  each  other,  as  well  as  with  caretakers.  In  research  we  do  this  to  benefit  the  lives  of  our  animals  and  in  turn  have  seen  better  research  results.    

14. Swinging  into  action;  A  look  into  enrichment  of  a  small  cynomolgus  macaque  colony.  Jessica  Howell,  Cincinnati  Children’s  Hospital,  Cincinnati,  Ohio    As  a  newly  introduced  husbandry  technician  into  the  care  of  a  macaque  colony  at  Cincinnati  Children's  Research  Foundation  everything  is  still  a  learning  experience.  Working  with  the  small  colony  of  five  female  cynomolgus  macaques  has  provided  many  examples  of  how  enrichment  is  beneficial;  both  socially  and  environmentally.  Enrichment  is  an  essential  part  of  our  daily  routine,  and  helps  provide  for  the  physiological  and  psychological  health  of  our  animals,  which,  in  return,  has  shown  to  enhance  research  results.    Our  goal  is  to  improve  their  lives,  because  they  help  to  improve  ours.    

15. Increased  Cage  Height  Does  Not  Impact  the  Well-­‐Being  of  Rabbits.  Kay  Stewart,  R.V.T.,  R.L.A.T.G.,  C.M.A.R.  ,  Danielle  Guilfoyle,  B.S.,  Whitney  Preisser,  B.S.  and    Mark  Suckow,  D.V.M.,  D.A.C.L.A.M.,  Freimann  Life  Science  Center-­‐University  of  Notre  Dame    In  the  eighth  edition  of  the  Guide  for  the  Care  and  Use  of  Laboratory  Animals  (the  Guide),  the  recommended  rabbit  cage  height  was  changed  from  fourteen  inches  to  sixteen  inches.  In  contrast,  the  majority  of  our  rabbit  cages,  purchased  in  1985  after  the  USDA  prescribed  rabbit  cage  size  requirements,  provide  fifteen  inches  of  interior  cage  height.    A  review  of  the  literature  failed  to  identify  published  data  that  support  an  advantage  to  rabbits  having  16  inches  of  cage  height  versus  14  or  15  inches.  The  study  described  here  evaluated  the  benefit  of  this  minimal  change  in  cage  height  for  rabbits  by  comparing  the  effect  of  the  cage  height  on  the  health,  growth,  and  overall  well-­‐being  of  the  rabbits.    Groups  of  ten  New  Zealand  white  rabbits  were  housed  in  cages  that  provided  either  15  inches  or  18  inches  of  interior  cage  height.    The  rabbits  were  observed  for  25  one-­‐hour  periods  over  seven  weeks  and  various  behavioral  parameters  scored.  In  addition,  rabbits  were  weighed  weekly  and  general  clinical  health  assessed.  After  four  weeks,  the  groups  were  switched  to  the  alternate  housing.  No  significant  differences  were  observed  in  body  weight  gain  or  behavioral  parameters  between  groups  housed  in  cages  of  different  heights,  nor  were  significant  differences  observed  in  groups  of  rabbits  when  moved  from  one  cage  type  to  the  other.    In  addition,  all  rabbits  remained  clinically  healthy  through  the  course  of  the  study.  These  results  demonstrate  that  minimal  changes  in  interior  cage  height  neither  benefit  nor  harm  rabbits.    

16. Pododermatitis  in  Rabbits  Housed  Long  Term  on  Wire  Bottom  Caging.  Manning,  Nicole*  Rogers,  Karen  L.  Allen,  Carolyn,  Cincinnati  Children’s  Hospital,  Cincinnati,  Ohio    In  a  colony  of  New  Zealand  White  rabbits  maintained  for  antibody  production  and  cardiovascular  studies,  several  of  the  long-­‐term  adult  rabbits  are  displaying  signs  indicative  of  ulcerative  pododermatitis,  or  “sore  hocks”.      Symptoms  include  alopecia  with  erythema,  hyperkeratosis,  ulcerations  with  scabs,  abscesses,  and  cellulitis  of  the  plantar  surface  of  the  proximal  metatarsus.      

2014  District  5  AALAS  Meeting  This  condition  has  been  noted  in  rabbits  weighing  4  to  6  kg,  housed  on  wire  bottom  flooring  and  of  at  least  1  year  of  age.    Thus  far  the,  condition  appears  independent  of  genetic  or  study  predisposition,  and  is  evident  to  be  caused  or  exacerbated  by  environmental  factors.    The  condition  seems  to  mimic  pododermatitis  in  rats  and  dogs  housed  on  wire  bottom  cages.    The  clinical  signs,  environmental  conditions,  staging,  pathophysiology  and  possible  treatment  options  will  be  explored  and  described.      

17. Bovine  Enrichment  in  a  Laboratory  Animal  Setting.  Regina  L.  Turner,  A.S.  and  Leslie  C.  Sherwood,  D.V.M.,  Cardiovascular  Innovation  Institute  and  Research  Resources  Facilities,  University  of  Louisville,  Louisville,  Kentucky    Cattle  are  not  commonly  used  laboratory  animal  species  and  when  housed  within  vivaria,  providing  stimulating  enrichment  can  be  a  challenge.    A  number  of  enrichment  strategies  and  manipulata  have  been  utilized  in  an  attempt  to  provide  adequate  environmental  enrichment  for  3-­‐6  month  old  Jersey  and  Holstein  calves  housed  within  our  facilities  over  a  period  of  7  years.  Effective  and  ineffective  strategies  and  manipulata  will  be  presented.  Discussion  regarding  effective  cleaning  and  sanitation  of  objects  will  be  included  as  well  as  stereotypical  behavior  observed  when  calves  are  not  provided  with  adequate  environmental  enrichment.    

18. Retro-­‐orbital  Injections  in  Mice.  Holly  Stallkamp,  R.V.T.,  R.L.A.T.G.,  S.R.T.,  University  of  Cincinnati,  Cincinnati,  Ohio    Retro-­‐orbital  injections  are  an  alternative  to  intravenous  tail  vein  injections.  They  are  technically  less  challenging  when  compared  to  tail  vein  injections.  Anesthesia  is  required  for  the  procedure.  Isoflurane  works  well  since  induction  and  recovery  time  is  minimal.    

19. Outbreak  of  Mycobacterium  liflandi    in  a  Colony  of  Xenopus  Leavis  Frogs.  Mary  Zabonik  B.A.,  A.A.S,  R.L.A.T.,    Laboratory  Animal  Technician,  Nationwide  Children’s  Hospital,  Columbus,  Ohio    Xenopus  leavis  (African  clawed  frog)  embryos  are  used  for  the  study  of  embryonic  development  and  specifically  neuronal  gene  activation  research  at  The  Research  Institute  at  Nationwide  Children’s  Hospital.  The  Animal  Resources  Core  houses  a  large  colony  of  female  and  male  Xenopus  leavis  frogs  in  a  flow-­‐through  Pharmacal  PharmHouse™  aquatic  tank  system  to  support  necessary  embryo  collection.    Over  a  several  month  period  of  time,  frogs  began  exhibiting  clinical  signs  ranging  from  cutaneous  erythema  and  ulcers,  edema,  coelomic  effusions,  whole  body  bloating,  lethargy,  and  death.  Tissue  samples  and  water  samples  were  submitted  for  testing.  Diagnosis  of  Mycobacteria  liflandi  was  confirmed  by  polymerase  chain  reaction.  This  poster  presents  the  steps  taken  to  diagnose  the  disease  in  our  colony  and  procedures  implemented  to  depopulate  the  colony  as  well  as  disinfect  equipment  prior  to  repopulation.    

20. Husbandry  Adaptations  for  the  containment  of  Hepatitis  A  and  Hepatitis  E  viruses  in  infected  Rhesus  Macaques.  Natalie  Snyder  and  Ellyn  Peterson,  Nationwide  Children’s  Hospital,  Columbus,  Ohio    Hepatitis  A  virus  (HAV)  and  Hepatitis  E  virus  (HEV)  were  used  to  infect  two  separate  groups  of  rhesus  macaques  as  part  of  an  immunological  study.  HAV  and  HEV  infected  macaques  were  housed  in  adjacent  rooms  with  in  an  ABSL-­‐2  facility.  Highly  contagious  nature  and  route  of  transmission  of  these  viruses  called  for  unique  adaptions  to  standard  primate  husbandry  procedures  including  methods  for  cage  cleaning,  providing  environmental  enrichment,  feeding  and  veterinary  procedures.  Due  to  the  unexpectedly  prolonged  course  of  viral  shedding,  further  adaptations  in  husbandry  practices  were  made  to  overcome  ergonomic  and  time  constraints.  The  details  of  these  husbandry  modifications  including  the  pros  and  cons  will  be  discussed  in  this  presentation.  

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