Laboratory Animal Handling Technique By A. Mallik & Dr.S Nayak

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Laboratory Animal Handling Technique:

Laboratory Animal Handling Technique Mr.Arunabha .Mallik Dr. S.Nayak Assistant Professor Principal & Professor Bansal College of Pharmacy Bansal College Of Pharmacy Dept.of Pharmacology

Objective:

2 Objective To provide basic concepts of animal handling technique to new animal user To comply with the Animal Welfare Ordinance and avoid mishandling of animal in research

Laboratory Animal Handling Technique - Rat:

3 Laboratory Animal Handling Technique - Rat Handling and Restraint Oral Feeding Subcutaneous injection Intraperitoneal injection Blood collection from tail vein

Handling and Restraint :

4 Handling and Restraint Initially restrain a rat, the handler should gently grasp it around the shoulders . ↓ The handler’s thumb can then be placed under the rat’s mandible, to prevent bites, ↓ The rat’s hind limbs can be supported with the other hand

Oral Feeding in Rat:

5 Oral Feeding in Rat Gavaging is used to dose an animal with a specified volume of material directly into its stomach. Only a specialized, commercially available gavage needle should be used for attempting this procedure. Fill the syringe with the appropriate volume of material and attach the needle. Place the tip or ball of the needle into the animal’s mouth. Slide the tip gently past the back of the tongue. The needle should slide easily down the esophagus, if properly placed. DO NOT FORCE!!! If any resistance is met, remove the needle and reinsert. Once the needle is properly placed, administer the material. Feeding amount limited to 1% of body weight

Tools for Oral Feeding in Rat:

6 Tools for Oral Feeding in Rat 16 G ball-tipped feeding needle with syringe Leather glove

PowerPoint Presentation:

7 Restrain the shoulders of the rat by your thumb and index finger, then support the lower limbs with your right hand

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8 Let the rat lying on your left palm and introduce the ball-tipped feeding needle from the pharynx in to the esophagus when the rat is in the act of swallowing

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9 The anatomy picture showed the position of the feeding needle tip inside the esophagus

Subcutaneous Injection in Rat:

10 Subcutaneous Injection in Rat Use your thumb and forefinger to make a tent of skin over the scruff. Prep the area with 70% ethanol. Insert the needle, bevel up, at the base of the tent. The needle should be inserted parallel to the skin and should be directed toward the posterior of the animal. Aspirate to ensure proper placement and inject the material.

Tools for Subcutaneous Injection in Rat:

11 Tools for Subcutaneous Injection in Rat 75% alcohol cotton ball for surface disinfection 25G 1 “ needle with 1 ml syringe for injection

PowerPoint Presentation:

12 Grasp the loose skin on the back of the rat 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 rat Inject the substance slowly

Intraperitoneal Injection in Rat:

13 Intraperitoneal Injection in Rat A common method of administering drugs to rats. Prep the site with 70% ethanol. The sterile needle should be placed. Insert the needle at a 30° angle. Aspirate to ensure proper placement and inject the material.

Tools for Intraperitoneal Injection in Rat:

14 Tools for Intraperitoneal Injection in Rat 75% alcohol cotton ball for surface disinfection Medium size towel for restraining the rat 25G 5/8” needle with 1cc syringe for injection

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15 Stretch the body of the rat by pulling up it’s tail and then cover the rat with a towel by your left hand

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16 Fold the skirts of towel under the rat from all directions

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17 The injection site should be in the lower left quadrant of the abdomen because vital organs are absent from this area

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18 Only the tip of the needle should penetrate the abdominal wall to prevent injection into the intestines .

Collection of blood from tail vein in Rat:

19 Collection of blood from tail vein in Rat General anesthesia needed small amount: 0.1-1 ml This method must be used with caution, as when improperly performed, permanent tail injury may occur

Tools for collection of blood from tail vein:

20 Tools for collection of blood from tail vein 75% alcohol cotton ball for surface disinfection 27G1/2” needle with 1 ml syringe for blood withdrawal a vial for blood collection

PowerPoint Presentation:

21 Optimal site of blood withdrawal is around the distal one-third of the tail since this part of tail gives better visualization of the veins

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22 Disinfect the tail with 75% alcoholic cotton ball

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23 When the needle penetrates the epithelium of the tail, pull back the plugger a bit to create negative pressure inside the syringe, then push the needle in the vein slowly until blood get into the dead space of the needle head

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24 Pull back the plugger by the ring finger to withdraw blood from the tail vein

Laboratory Animal Handling Technique - Mouse:

25 Laboratory Animal Handling Technique - Mouse Oral Feeding Intraperitoneal injection Blood collection from Orbital Sinus in Mouse

Oral Feeding in Mouse:

26 Oral Feeding in Mouse Gastric intubation ensures that all the material was administered Feeding amount limited to 1% of body weight

Tools for Oral Feeding in Mouse:

27 Tools for Oral Feeding in Mouse A 18 G stainless steel, ball tipped needle A glove

PowerPoint Presentation:

28 Grasp the loose skin on the back of the mouse and restrain it ’ s tail with your ring finger and little finger. Then, introduce the feeding tube from the pharynx in to the esophagus when the mouse is in the act of swallowing.

Tools for Intraperitoneal Injection in Mouse:

29 Tools for Intraperitoneal Injection in Mouse 75% alcohol cotton ball for surface disinfection 25G 1/2” needle with 1 ml syringe for injection

Place a mouse on a cage lid and grasping the loose skin behind the ears with your thumb and forefinger :

30 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 :

31 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

PowerPoint Presentation:

32 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 Orbital Sinus in Mouse:

33 Blood Collection From Orbital Sinus in Mouse Should apply anesthetic before blood withdraw A convenience and easy apply method for blood collection in mouse Collect amount up to 0.5 ml

Tools for Blood Collection from Orbital Sinus in Mouse:

34 Tools for Blood Collection from Orbital Sinus in Mouse 75% alcohol cotton ball for surface disinfection General anesthetic 27 G needle with 1 ml syringe for injection Glass capillary tube and vial for blood collection

Anesthetize a mouse by intraperitoneal injection :

35 Anesthetize a mouse by intraperitoneal injection

Use a sharp end glass capillary tube to penetrate the orbital conjunctiva and rupture the orbital sinus :

36 Use a sharp end glass capillary tube to penetrate the orbital conjunctiva and rupture the orbital sinus

Collect blood with a vial :

37 Collect blood with a vial

PowerPoint Presentation:

38 THANK YOU Mr.Arunabha .Mallik Dr. S.Nayak Assistant Professor Principal & Professor Bansal College of Pharmacy Bansal College Of Pharmacy Dept.of Pharmacology

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