MEENAKSHIM.Sc. 1st Year

ANIMAL HANDLING TECHNIQUES

OBJECTIVESTo provide basic concept of animal handling

to new user.To comply with the Animal Welfare

Ordinance and avoid mishandling of animals in research.

Handling means petting, feeding and watering, cleaning, manipulating, loading, crating, shifting, transferring, immobilizing, restraining, treating, training, working and moving, or any similar activity  with respect to any animal.

Social Housing  Nonsocial enrichment- The room The cage  Food Disease free: weight

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

Handling and restrainInitially 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.

Oral feeding in mouseTOOLS FOR ORAL FEEDING

A 18 G stainless steel, ball tipped needlea glove

introduce the feeding tube from the pharynx in to the esophagus when the mouse is in the act of swallowing.

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

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.

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 the orbital sinus

Collect blood with a vial

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

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

Laboratory Animal Handling Technique - RabbitA. Intramuscular injectionB. Subcutaneous injectionC. Blood collection from ear veinD. Blood collection from carotid artery

Intramuscular Injection in Rabbit

Subcutaneous Injection in RabbitDisinfect 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 needle .

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

Tools for collecting 50ml blood from ear artery

75% alcohol cotton ball for disinfect of ear surface21 G winged infusion set with 50 ml syringe for blood collectionDry 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

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

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

THANK YOU