Neil S.

Lipman, VMD
Memorial Sloan-Kettering Cancer Center
Weill Cornell Medical College

Program Overview
Pros and Cons: Engineering vs.
Performance Standards
Intra-program Standard Development
Examples
Conflicting Standards
Looking Forward

No typical academic program

Bi-Institutional
Mouse-centric
Harmonized
Laboratory support
Postdoctoral trainees

Ensures a minimal
standard the floor
Necessary
Standardization
Consistency
Ideally based on objective
data
Implementation
easier (B and W)
less costly (+/-)

Can be empirical

anthropomorphic
political influence

Problematic if based on
poor data
If tightly defined
restricts process
retards new knowledge

Difficult to
change/inflexible
Can result in unnecessary
costs
Lack of transparency

Greater flexibility
Not static
Alignment with science
Improve animal welfare
and care
Tailorable
Can result in the
refinement and
improvement of
engineering standards

Cost/time of developing
Subjectivity
Differing views on desired
outcome
Difficult to determine
what to measure
Challenging for programs
that are resource limited

Reframe: Evidence based or outcome driven

decision making and problem solving
Management approach/style
Engineering standards while recognized not
emphasized
Hypothesis and data driven when possible
Bias recognized & managed
Objective vs. subjective process
Transparent
Outcome sharing = publication

animal health/welfare

customer
service

science

Limited methods available to reliably and permanently

identify pre-weanling mice
Toe-clipping used at MSKCC/WCMC in preweanling
mice following institutional guidelines
Toe-clipping: strong anthropomorphic views expressed
BVAAWF/FRAME/RSPCA/UFAW Joint Working
Group on Refinement and Production of GEM (2003):
should not be routinely used
assumed to cause pain & impact animals ability to grip
last resort method
performed under local anesthesia

Limited objective published information evaluating

effects prior to 2010
Guide 7th ed Toe-clipping, as a method of identification of small
rodents, should be performed when no other individual identification
method is feasible and should be performed only on altricial
neonates.
Guide 8th ed Toe-clipping should only be used when no other
individual identification method is feasible. It may be the preferred
method for neonatal mice up to 7 days of age as it appears to have
few adverse effects on behavior and well-being at this age,.(refs)

Desired Outcomes
Minimize pain and distress
Limit short and long-term physiologic and behavioral
effects
Safe for animal and operator
Fast and easy
Permanent
Unique number to be assigned to a large number of mice
Option of simultaneous tail biopsy
Low cost

4 groups evaluated at PND 7 and 17

1. toe-clip: no anesthesia
2. toe-clip: topical vapocoolant
3. sham: no anesthesia
4. sham: topical vapocoolant

C57BL/6 pups
Experienced technician
Sharp, microsurgery scissors, wiped
2 toes clipped at level of the proximal
phalange:
digit 3 on right forepaw
digit 3 on left hindpaw

Pups observed for immediate reaction

& @ 1, 3, 5, and 12 h

Open Field Test

Modified
SHIRPA &
grip test
Beam Test
Rotarod

Daily handling
Elevated Plus
Maze
Week 7

Week 8

Three 60s
trials/day X3 days

Week 9

Euthanasia by CO2
Harvest of toes for
histology

Week 10

Signs of distress

Toe-clipping

Vapocoolant + Toeclipping

Vapocoolant

PND 7: ~60%
vocalized, d activity

PND 7: dragging of sprayed limb post-clip

PND 17: little reaction

PND 17: vocalization, urination, limb

withdrawal

Bleeding

PND 7 & 17:

immediate clotting

PND 7 & 17: Delayed

profuse bleeding

Ease of procedure

PND 7: Difficult to clip

small toe size

PND 7 & 17: VC causes toes to stick together

hard to isolate toe clipping difficult

Post-procedural
observations

No maternal rejection

No maternal rejection

No maternal rejection

PND7: swelling of HP
12 h

PND 7 & 17: swelling

of paws 12 h

PND 7: swelling of
paws 5 h

PND 17: no
abnormalities

N/A

PND 17: swelling of

paws 12 h

No developmental
differences
No behavioral
differences
No differences in
weight gain
Digit fully innervated at
PND 7
Sig. diaphyseal
ossification by PND 7

PND 7

PND 7

Outcome
Revised institutional guidelines
Deleted recommendation for
vapocoolant topical anesthesia
Extended recommended ages < PND 17
Limit to 1 digit per paw for ID using
recmd numbering system
Toe-clip site distal 1/3rd of 1st phalanx
PND 14 -17: toe-clip and tail biopsy when both
necessary

Paluch et al (2014) JAALAS 53:132-140

Circa 2000
Large numbers of mice of
incorrect genotype require
euthanasia
Labor intensive
Few and imperfect systems
and solutions
commercial
home grown

Desired Outcomes
Achieve rapid
unconsciousness and
death
Avoid excitement
Minimizes fear, distress,
anxiety
Reliable
Simple to administer
Reproducible
As specified by the AVMA, ACLAM
and FELASA

Safe for the operator

Aesthetically acceptable to
the operator
Minimize changes in the
animals environment
Remain in home cage
Visualization
Ensures death vs narcosis
Minimize labor/efficient
large #s simultaneously
low cost

Outcome
Automated, logic controlled
system
Based on individually
ventilated caging system
Fully validated and tested
Videography behavioral
assessment
Up to 80 cages per cycle
Operate 15 systems > 9 years
McIntyre et al (2007) JAALAS
46:65-73

AVMA Guidelines for the

Euthanasia of Animals: 2013
Edition
CO2 General recommendations:
a displacement rate from 10 30%
of the chamber volume/min is
recommended.
Small Laboratory and Wild-Caught
Rodents (CO2): An optimal flow rate
for CO2 euthanasia systems should
displace 10% to 30% of the chamber
or cage volume/min.

Resources ($$$)
Extramural funding essential
Consortia
Multi-Site
Example: space and cage size