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Mouse
• Popular model organism in mammalian belonging to the Mus musculus
species

Anatomy, Physiology, Restraining, • Mouse owes small body size and high fecundity

Blood collection and Compounding of drugs • Head: small, triangular, small relative to body

• Ears are large relative to the head

Mouse has thin tails where length of tail is longer than body
Legs of mice are small relative to body

Mice Rat
• single species Norway rat (Rattus norvegicus)
• 40 chromosomes in their diploid genome • Wistar, Fisher and Sprague Dawley
• Wide head, long ears
• outbred (Swiss albino) Vs inbred (Balb c and C57BL/6J)

• Athymic mice

Uses • substance abuse,

• Obesity model • anxiety,
• Tail (thick) length that is always lesser than its body length
• heart disease, • aging,
• Muzzle: large and blunt with wide muzzle
• diabetes, • cancer
• Legs (specially hind legs) are comparatively longer relative to body
• arthritis, • Parkinson disease • Ears are small relative to the head

Rabbit
• Newzeland White Rabbits (Oryctolagus cuniculus)
• 42 diploid numbers of chromosomes • The bones are relatively fragile

• Wistar rats are an outbred first strain: Wistar Institute in 1906

• Sprague Dawley rat (Calm rat) and Long-Evans rat strains

were developed from Wistar rats

• Zucker rats: obesity research

•well-developed nictitating membrane, the third eyelid.

•four pairs of mammary glands.
•Large Eyes, Large prominent Ear veins & Large Ova.
•highly vascularized ears are important in thermo- regulation.
•Kids born with eyes & ears closed.
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Animal restraint

Rat may be palmed over its back with the index and middle finger placed between its
mandible to minimize the chance of being bitten.

Rabbits should never be grasped and carried by the ears.

RABBIT
Improperly held rabbits may inflict scratches on the handler with

• Rabbits are properly restrained to ensure the safety of the its rear limb.

animal and the handler. Restrained only for minimum amount of time needed to perform
any experimental manipulation.
The relative strength of the hind
The rear quarters of the animal should be supported during
limbs of the rabbit to the
restrain.
fragility of the skeleton may
translate into a fracture of the
lumbar spine if the rabbit’s
weight is not supported
properly.
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Sexing Mice and Rat Sexing Rabbit

Rabbits can be sexed by causing eversion of the penis or vulva when
The male and female mice and rat can be differentiated by slight pressure is applied to the external genitalia. The presence of
observing the distance from the anus and genital papilla which is scrotal sacs in males and mammary glands (5 pairs) in females can
greater in males. also be noted.

• Orbital plexus
Routes of Blood collection in Rat & Mice • Penetrating the retro-orbital sinus in mice or plexus in rats with a glass capillary
tube or Pasteur pipette.
• Tail vein
• Animal should be anesthetized with the collector’s non-dominant thumb and
• Drawing blood from the lateral tail vein index finger used to hold the rat’s head steady.
• With gentle caudal traction the eye will protrude and the thumb occludes the
• source which will dilate the vasculature. jugular vein. A capillary tube or pipette is positioned at the medial canthus
• rat is held in a manual restraining device. An (inner corner) of the eye and the ocular conjunctiva and the underlying orbital
plexus entered caudomedially.
assistant can hold the tail steady during • Apply a small quantity of ophthalmic ointment on the eye, to aid hemostasis
venipuncture and reduce the incidence of infection.

• vessel can be entered with a 21-gauge or smaller

needle.

• If serial blood collections are desired, the first

sample should be taken as distal as possible
(close to the tip of the tail) and subsequent
samples taken proximally.

Facial vein Saphenous vein

• Blood collection from the • This method used if multiple samples are
submandibular facial vein taken in the course of a day.

• approximately 200ul of blood can be • saphenous vein is seen on the surface of the
obtained thigh.

• Materials needed include a 20 or 22 G • Use a 24 G needle to puncture the vein and

hypodermic needle, blood collection tubes release blood from the saphenous vein.
and sterile gauze. • approximately 100 microliters can be
collected.

• A cotton ball is applied to the puncture site to

stop further bleeding.
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Cardiac puncture
Routes of Blood collection in rabbit
Marginal ear vein
• Done only as a terminal procedure in an anesthetized mouse/ rat.
• The marginal ear vein of the rabbit is useful for
• method requires training and skill, but can be used to obtain
comparatively large volumes of blood. collection of small volumes of blood
24-gauge needle is either inserted
• held in a restrainer or else should be sedated.
through the diaphragm lateral to the
• area is shaved and cleaned with alcohol.
xiphoid cartilage and directed
• Use of a butterfly set may avoid damage to the
forward and medially towards the
vessel if the animal moves.
heart; or inserted between the fifth
Central ear artery
and sixth ribs on the left side and
• used for collecting larger volumes of blood
directed forward toward the heart.
• 10 ml of blood/kg body weight can be collected
Decapitation
used to obtain comparatively large volumes of blood, • 20 or 21 gauge needle pointed towards the base
of the ear.

Cardiocentesis
• acceptable method of blood collection from rabbits when used as a
Blood withdrawal frequency
terminal procedure • Total blood volume = 66 ml/kg
• not suitable as a repetitive blood sampling procedure since it • Maximum blood withdrawal in 2 weeks
carries considerable risk to the animal without fluid replacement = 10% of
• rabbit should be at a surgical plane of anesthesia before total blood volume
performing cardiocentesis • Maximum blood withdrawal 2 weeks
• point of maximum intensity of heartbeat should be determined with fluid replacement = 15% of total
blood volume
• 18 gauge needle attached to a vacuum tube should be used
• Daily blood withdrawal for 2 weeks
without fluid replacement = 1% of total
blood volume

Intramuscular (IM)

Routes of compound administration • Caudal hind limb muscles of lateral thigh are a common site for IM
administration in the rat, mice and rabbit.
• Injection should not be made too deeply, as
it is important to avoid the femoral nerve
• Intramuscular
and hitting bone.
• Intravenous
Intravenous (IV)
• Subcutaneous • Typically a small quantity of blood will “flash” back into the hub of the
needle if properly positioned in the vein.
• Intradermal • Rat, mice – Tail vein; Rabbit – Marginal ear vein
• Hair over the vein is clipped or shaved and the skin
• Intraperitoneal cleansed with alcohol or alcohol-iodine before
making the injection.
• Per os • Vein distended by flicking the margin with the
• fingers a few times.
• Xylene may be used as a vasodilator by gently rubbing a small amount of it
over the outer surface of the ear.
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Subcutaneous (SC/SQ) Intraperitoneal (IP)

• Tenting the skin and inserting a needle Injection into the peritoneum to the animal’s lower abdominal quadrant to
between the skin and underlying tissues. avoid vital organs.
• If nothing is aspirated into the syringe, the needle should enter the abdomen at a shallow angle, and once into
injection is then made under the skin over
the back and sides peritoneum the investigator should gently aspirate.
• And between the scapulae. If one aspirates intestinal contents following placement of the needle into the
peritoneum, dispose of the needle, syringe, and solution due to
Intradermal (ID) contamination.
• Given into shaved areas over the thorax Administering compounds IP results in their absorption into the hepatic
and abdomen. circulation prior to their distribution to other organs.
• One should position the needle at an Caution must be taken to avoid puncturing a distended urinary bladder, the
acute angle to the skin which is held bowel, or the liver.
taught so the bevel of the 23-gauge or
smaller needle enters only the skin and
not the subcutaneous tissue.
• One may ensure proper placement upon
• injection when a small bleb forms.

GUIDELINES FOR COMPOUND ADMINISTRATION

Gastric Gavage/Per Os (PO) Notes
Route Volume to be administered
• Bulb-tipped gastric gavage needle.
Rat Mice Rabbit
• Length of the needle should approximate the distance from the mouth to Administer volume in 2–3 sites
SQ 5 mL/kg 2-3mL 30-50 mL
just beyond the last rib. Vasodilate the vein and one should
IV Max 0.5 mL Max 0.2 mL Max. 5 mL
• Firmly restrain the animal and pass the needle through the side of the start distally (farthest from the
heart) and move proximally
mouth and advance the needle towards the esophagus.
IM 0.1–0.2 mL/site 0.05mL 0.5 mL Volume should be restricted as the
• The animal will swallow; ensure that the needle is in the stomach rather muscle mass is small and avoid
than the lungs. A deep penetration to prevent the
danger of damaging vital structures
• dministration of a small amount of the substance without vocalization or Warm fluids to body temperature;
IP 10 mL/kg 2mL 50-100 mL
struggling indicates proper positioning. administer over 2-3 min; single
injection volume should not exceed
10% of the circulating volume;
continuous 24h infusion should be
0.4 ml/kg/h

ID 0.05 mL/site Proper placement so that the

needle enter only the skin.
Gastric gavage 20 mL/kg Care must be taken that the tube or
needle does not enter the trachea
or puncture the esophagus or
stomach