This document provides information on proper animal handling techniques. It discusses objectives of avoiding mishandling and complying with regulations. Procedures for mice include oral feeding, injections, and blood collection from various sites. Rabbit techniques include intramuscular injection, blood collection from ears or arteries. Precise restraint and use of the correct tools are emphasized to safely perform common procedures.

1. MEENAKSHI
M.Sc. 1st Year
ANIMAL HANDLING
TECHNIQUES

2. OBJECTIVES
 To provide basic concept of animal handling to
new user.
 To comply with the Animal Welfare Ordinance
and avoid mishandling of animals in research.

3.  Handling means petting, feeding and watering, cleanin
g, manipulating, loading, crating, shifting, transferring,
immobilizing, restraining, treating, training, working an
d moving, or any similar activity with respect to any a
nimal.
Social Housing
Nonsocial enrichment-
 The room
 The cage
 Food
 Disease free: weight

4. Laboratory animal handling
technique- Mouse
 Handling and restrain
 Oral feeding
 Subcutaneous injection
 Intraperitoneal injection
 Blood collection from tail vein
 Blood collection from orbital sinus
 Blood collection from cardiac puncture

5. Handling and restrain
 Initially restrain a mouse, handler should gently grasp it
around the shoulder.
 Handlers thumb can be placed under mandible, to
prevent bites.
 hind limb can be supported with other hand.

6. Oral feeding in mouse
 TOOLS FOR ORAL FEEDING
A 18 G stainless steel, ball
tipped needle
a glove
introduce the feeding tube from the
pharynx in to the esophagus when
the mouse is in the act of swallowing.

7. Subcutaneous Injection in Mouse
Pick up a nude mouse and spin
it’s tail to put it in a faint
condition
Grasp the loose skin on the back of the
mouse from ears along the legs and
restrain the legs with your ring finger and
little finger
After disinfect the surface area,
insert the needle in the lateral side
of the abdominal wall and push
upwards to the armpit of the
mouse
A lump of injection substance can be
seen through the skin after injection

8. Intraperitoneal Injection in Mouse
Place a mouse on a cage lid
and grasping the loose skin
behind the ears with your
thumb and forefinger
As soon as the mouse’s head is
restrained, the mouse can be picked up
and the tail secured within your ring
finger and little finger
The injection site should be in the lower left quadrant of the abdomen because
vital organs are absent from this area. Only the tip of the needle should
penetrate the abdominal wall to prevent injection into the intestine.

9. Blood Collection From Tail in Mouse
Tools for Blood Collection from Orbital Sinus in Mouse
Push the mouse into the
restrainer
Amputate the tip of the
mouse tail by scissors
Massage the tail and
collect blood by
pipetteman
Anesthetize a mouse by
intraperitoneal injection of
Hypnorm
Use a sharp end glass
capillary tube to penetrate the
orbital conjunctiva and rupture
Collect blood with a
vial

10. Blood Collection From Cardiac Puncture in
Mouse
Anesthetize a mouse by intra-
peritoneal injection of Hypnorm
Disinfect the thorax area
with 75% alcohol cotton
ball
Insert a 24G 1” needle through the
thoracic wall at the point of maximum
heart palpitation
Search for the maximum
heart palpitation with
your finger
Withdraw blood slowly by your right
hand

11. Blood Collection From Saphenous Vein in Mouse
Pull out the leg and
removed the hair by a
assistant
Hair can also be shaved
by using a small scalpel
Apply vaseline after
disinfect the surface area
to reduce clotting and
coagulation during blood
collection.
Use a 24 G 1” needle to
puncture the vein and
release blood from the
saphenous vein
Use a Microvette or a
pipetteman with tip to
collect blood from the
saphenous vein
Approximate 100 microliters
can be collected
Flex the foot of the
mouse to reduce the flow
of blood back to the
puncture site
A cotton ball is applied
to the puncture site to
stop further bleeding

12. Laboratory Animal Handling Technique -
Rabbit
 A. Intramuscular injection
 B. Subcutaneous injection
 C. Blood collection from ear vein
 D. Blood collection from carotid artery

13. Intramuscular Injection in Rabbit
Subcutaneous Injection in Rabbit
Disinfect of injection site by 75%
alcohol directly or 75% alcohol
cotton ball
Pinch the skin up and insert the
needle in for injection
Disinfect of injection site
by 75% alcohol directly or
75% alcohol cotton ball
Pinch the skin up and insert
the needle in for injection
Keep the needle tip in skin for a
while after injection to prevent
leakage of substance from the

14. Collection of Blood from Ear Vein in
Rabbit
Ear vein before
administering of
tranquilizer
Ear vein after
administering of
tranquilizer
Insert needle into the ear
vein with needle tip surface
face up
Fix the position of needle
by your left thumb and
withdraw blood from the
ear vein
Press a cotton ball on the
top of the injection site
before pull out the needle.
Apply clip on the top of
the cotton ball for 2-3
minutes to stop further
bleeding

15. Tools for collecting 50ml blood from ear
artery
75% alcohol cotton ball
for disinfect of ear
surface
21 G winged infusion set
with 50 ml syringe for
blood collection
Dry cotton ball and
plastic clip for prevention
of further bleeding
Apply same procedure as
collecting blood from ear vein for
blood collection from ear artery

16. Blood collection from carotid artery
Restrain the anesthetized
rabbit on the surgical table
Remove the hair
around the neck area
Make a 5 cm opening on
the middle line of the
neck
Search the carotid artery
with a tissue forceps and a
homeostatic forceps.
Tools for blood collection
from carotid artery
Isolate the carotid artery from
the vagus nerve and other
connective tissue

17. Tie up the distal end of the
carotid artery by silk suture
Clip the proximal end of the
carotid artery with a bulldog
clamp
Insert a 16 G
intravenous catheter
into the artery
Fix the position of the catheter tip
by two silk suture knots
Release the bulldog clamp and
collect blood from the artery though
the catheter into the centrifuge tube