Name: Date

Group No.

LEARNING ACTIVITY NO. 4

PROPER ANIMAL HANDLING AND MANIPULATION

OBJECTIVE(S):

1. Determine the different sites of administration.

2. Identify the recommended needle sizes for different test animals.
3. Perform the proper blood collection in mice, rat and rabbit.
4. Apply the proper techniques in caring and handling test animals.

Laboratory animals are used for the experimentation and for toxicity testing of
chemical substances because society has determined that man must not be used.
Although it is the basis of much discussion, there seems to be a little doubt that the
results from laboratory animals can be extrapolated successfully to man. This has been
particularly emphasized by retrospective clinical studies in a man in which the toxic side
effects of drugs observed in animal are compared to those found initially in animal
studies.
The skill of handling laboratory animals will not be acquired easily. It may take many
hours of practice on just one species to become professional. Observations of the
points listed below will help achieve this.:

1. Try to overcome fear or anxiety for it is almost impossible to work under these
conditions. If handler is not relaxed then neither will the animal be.
2. Incorrect handling of animals may cause physical damage and will certainly lead to
stress, all of which will render them unsuitable for experimentation.
3. Newer subject the animal to any procedure until it has been successfully relaxed and
restrained.
MOUSE
Healthy, randomized and properly identified adult mice, each weighing 17 to 30
grams at the start of the experiment are to be used.
It must be stated that the one unfortunate problem with handling mice is they will
almost certainly try to bite the operator. After many weeks of handling they will become
reasonably tame but even they cannot be trusted. Certain strains are more
temperamental than other. Care must be taken not to excite the mice before opening
the cage, avoid sharp movements and high-pitched noises such as whistling. If possible
slide the cage lid back a fraction or just enough to get a hand in; a mouse can jump
several feet into the air, but rarely becomes injured as a result of a fall.

Handling and Restraint

a. Using right hand, catch or pick up the mouse by the tail about 1 inch from the rump
and place it on a surface where it can grip with all four feet but keep hold of the
tail. The best surface for this purpose is the wire lid of the cage. Placing the mouse
across the mesh so that it can grip with all four feet will ensure that it remains still
long enough to be taken by the scruff. If the mouse attempts to turn back the tail
must be pulled gently and it will go forward again.
b. Then while on this position pick the mouse using the thumb and forefinger of the
left hand on the scruff. This must be carried out quickly and deliberately on
one movement. Care must be taken to gather enough of the loose skin at the back
of the animal’s neck for it is able to turn its head, it will certainly bite! When this
happens, do not pull the mouse away from the bitten finger! Release mouse’s tail
and it will release its bite too.
c. When the mouse is scruffed the tail can then be held against the base of the thumb
by the rest if the fingers.
d. The animal is now ready for oral and parenteral administrations.
How to pick up a mouse
When you handle mice, all your movements should be slow and gentle. They are
lively animals, but they will rarely bite unless they are afraid or mishandled. Grasp the
middle of the tail between your thumb and index finger, and lift the mouse.
NOTE: Frequently, the animal will hang quietly and can be carried in this manner.
Occasionally, an individual will turn around and crawl onto the palm of your hand. It
should be allowed to do this.

How to pick up a mouse with thump forceps

The thumb forceps are shaped somewhat like eyebrow tweezer. The tips must be
covered with rubber or polyethylene tubing to prevent damage to the mouse’s tail.
Grasp the middle of the tail between your thumb and index finger, and lift the mouse.
CATION: too much pressure may cause pain or even crush the tail.

One-handed method
Follow the above steps
Then, place the mouse’s tail between the last two fingers of the hand that is holding the
nape.
Two-handed method
Place the mouse on a rough surface while holding the tail firmly.
NOTE: smooth surface will frighten the mouse because it cannot get a foothold. This
may cause it to turn around and bite in its attempt to escape. Grasp the nape gently and
firmly with your free hand and lift the mouse.

Administration of drugs and other substances.

1. Be sure to reduce discomfort and distress to the animal since stress and pain are
known to alter body responses to a wide range of stimuli.
2. Recommended sizes of needles and volumes for injection for mouse:
 Subcutaneous- scruff, 0.5 ml, <20G
 Intramuscular- Quadriceps/posterior thigh, 0.05 ml , <21G
 Intraperitoneal - 0.5 ml, <21G
  Intravenous- lateral vein, 0.3 ml (slowly), <25G
3. Oral dosing using a stomach tube or gavage tube.
The mouse should be scuffed ensuring that the head and neck are extended and
in line with the back. The tube then be passed into the mouth via the interdental
space and advanced gently rotating the tube to ease its passage into the
esophagus and or the stomach if necessary. A coughing mouse indicates that
the trachea is inadvertently intubated.
4. Subcutaneous injection. The injection is made on the area of the skin “tented’ by
scruffing of the restrained mouse. The needle should be positioned and almost
parallel to the skin and introduced through the skin.
5. Intraperitoneal injection
The needle is advanced parallel to the line of the leg, and pushed through the
abdominal wall into the peritoneal cavity. Injection is made on an area anteriorly.
Mice frequently urinate when restrained, and this should not be thought as a
result of the injection
6. Intramuscular injection
The muscles of the posterior or anterior thigh are the sites commonly used.
7. Intravenous injection
IV injections are carried out via the lateral tail vein. A loss of resistance should
be noticed as the needle enters the vein. Injection of a small volume of fluid will
indicate whether the needle is correctly positioned. Upon withdrawal of the needle,
firm pressure on the puncture wound for about a minute should be sufficient to
prevent any hemorrhage.

RATS
Healthy, randomized, properly identified rats with weights between 150-300 grams are
to be used. nOne or two strains of laboratory rat will possess a very placid temperant.
Even when upset by various treatments or not, being handled regularly for longer
periods, rats can be calmed and relaxed within a short time by correct handling.
Handling and Restraint
1. The adult rat should not be picked up by the tail as this can cause stress to the
animal and is not at all necessary.
2. Rats should be grasped firmly around the shoulders, the fingers winding round
the abdomen. The animal should then be turned over onto its back and the
thumb place under its chin. Animals that appear to be struggling at this stage may
be gently rocked backwards and forwards; this will have a calming effect.
3. Do not grasp the rat too much as it will impair respiration nor to loosely for the rat
will struggle and scratch because it feels unsafe. Hold the rump if the rat is large
i.e. over 250 grams.
4. Rats should never be handed from one person to another, as it is likely that they
will scratch during the transfer.
5. Before performing any procedures such as injections , oral dosing, etc., it is
desirable to restrain the rat as this is the time it will bite and scratch, the latter
being the most common injury to unskilled operators.
6. Pick up the rat with the right hand, leaving the head and shoulders slightly
protruding, then with the left hand and scruff should be taken between the thumb
and index finger. The rest of the fingers can then catch all the loose fur and skin
down the back to the base of the tail.
7. The rat should then be turned over onto its back to present a perfect target area
for injections. While in this position the rat is not only immobilized but also very
relaxed.

HOW TO HOLD A RAT FOR TREATMENT

Pick up the rat. Rotate the wrist of your right hand to expose the mid-section. Extend the
rat’s hind legs with your left hand, grasping one hind leg between your thumb and index
finger and the other between your index and second fingers.

Administration of drugs and other substances

1. Recommended size of needles and volumes for injections:
 Subcutaneous - scruff , back , 1-2ml, <20G
  Intramuscular - Quadriceps/posterior thigh, 0.2 ml, <20G
 Intraperitoneal - 2-3 ml, <21G
 Intravenous - Lateral vein, 1.0 ml (slowly), <23G
2. Procedure on administration of drugs is the same with the mouse.

RABBITS
Rabbits are very susceptible to lumbar spinal luxation, resulting in paralysis. It is
necessary to support the animal’s hindquarter at all times. Although rabbits seldom bite,
they can inflict painful scratches with their hind legs. One way of lifting a rabbit is by
grasping the skin over the shoulder with one hand and gently lifting it with the other. Arm
cradling the body, the head nestled in the crook of your arm. Rabbits must never by
lifted by the ears.

HOW TO REMOVE A RABBIT FROM A CAGE

Approach the cage and open the door slowly so that you will not startle the rabbit.
Reach in and grip the loose skin over the shoulders firmly but gently with your left hand.
Place your right hand under the rabbit’s rump to support its weight.
NOTE: the right hand may be opened and placed flat under the rabbit’s stomach
instead of under the rump, but this requires more skill, NEVER use the flat-handed
method for handling pregnant does. It might injure the unborn young. Lift the rabbit out
of the cage.

HOW TO CARRY A RABBIT

Grasp the loose skin over the neck with your left hand and place your right hand under
the animal’s rump, as you did in removing the rabbit from its cage. Pick up the rabbit
and hold it in an upright position.
CAUTION: never lift or carry a rabbit by its ears. They aren’t handles; Don’t carry a
rabbit by the neck skin alone. This is painful to the animal and will cause it to struggle.
You run the chance of getting scratched by the sharp rear claws. Carry the rabbit out in
front of yourself.
Subcutaneous injections
SC injections are easily performed in rabbits because of the laxity of their skin and the
large area into which material can be administered. The technique is the same as
described for mice, however injections should not be administered over the neck as this
is the site from which the animal is picked up. A 10- 22 gauge or larger gauge needle is
recommended. Volumes should not exceed 5 ml per site unless isotonic fluids are
administered. Approximately 30 ml of isotonic fluids can be administered per site and
multiple sites may be used. Always aspirate before injection.

Intramuscular injections
The recommended sites and technique for IM injection in the rabbit are as described for
rats. IM injection sites are illustrated in. However, the rabbit’s larger muscle mass
requires a longer needle (10) to adequately introduce the material deep within the
muscle belly, a distance of approximately 7-10 mm is an adult rabbit and acceptable
volumes to be injected are larger (< 1.5 mls.)

Intravenous injections
IV injections are straightforward in rabbits because of the ease of vascular access. The
marginal ear veins located on the lateral aspect of the rabbit’s ear are readily visible.
The veins can be made more prominent by occluding the vessel at the base of the ear
by gently holding off with your fingers or applying a small paperclip. The vein is
swabbed with an alcohol soaked cotton pledget. The needle is inserted as described for
injection rodent tail veins. Remember to aspirate to verify placement of the needle within
the vein. Remove the paperclip or your occluding finger prior to injection. Volumes of
approximately 5 to 10 ml can be administered if given slowly, however routine volumes
are frequently < 1 ml. A butterfly needle or a louer lock needle catheter may be inserted
if larger volumes are frequently < 1 ml. A butterfly needle or a louer lock needle catheter
may be inserted if larger volumes are to be administered or repetitive injections will be
given. A 0.5 24 ga or larger gauge needle is recommended. Pressure should be applied
over the injection site for approximately a minute to prevent hematoma formation.
Intradermal Injections
ID injections may be used to immunize rabbits. The technique is as described for guinea
pigs with following changes. The neck and anterior thoracic region should be avoided
for injection as rabbits are handled by grasping this region. Because of their larger size
up to 12 sites can be used.

PRACTICAL EXAM: Proper handling of mouse, rat and rabbit.

CONCLUSION(S):