Techniques in experimental

Pharmacology & Animal Handling.

From 16-1-2014 to 18-1-2014Venue- AV Room/ PG Research Lab

Department of PharmacologyPES College of Pharmacy

Topics

1.Importance of animal experimentation in biomedical research

2.Demonstration- Restraining, routes of drug administration

and blood withdrawal techniques in rats

Date- 16-1-2014 Time 9AM to 12 N.

Dr Shivalinge Gowda KPAsst Professor and HOD

1.Importance of animal experimentation in

biomedical research

In mouse 90% of the genes are similar to human beings. Hencethey become the effective model for the human body.

Animals suffer from similar diseases to

human including cancers, TB, flu and

asthma.

Nearly every Nobel Laureate in

Physiology since 1901 has relied on

animal date for their research.

Most of the animal and human drugs are same

Important drugs like anesthetics, the tetanus

vaccine, penicillin and insulin all relied on

animal research in their development.

Scanning techniques like CT and MRI were developed using animals

Herceptin- Largest selling drugfor the breast cancer isobtained from the mice

Tamoxifen

Household cats kill approximately 5 million

animals every week- more than the total

number of animals used in biomedical research.

Dogs, cats and primatesaltogether account for lessthan 0.2% of research animals.

More than 95% of research isdone on mice, rats, fish andbirds.

2.Rat handling and restraint techniques

3.Routes of drug administration in rats

a.Intra muscular route (IM)

Requirements-Disposable gloves, injection article,Isopropyl alcohol, gauge,1ml syringe, hypodermicneedle- 22-30G Max- 0.3 ml can be administered

3.Routes of drug administration in rats

b.Subcutaneous injection(SC)

Requirements-Disposable gloves ,injection article,Isopropyl alcohol, gauge,1-3ml syringe, hypodermicneedle- 22-30G Max -5ml can be administered

3.Routes of drug administration in ratsc.Intradermal injection(ID)

Requirements- Anesthetic, Disposable gloves ,injectionarticle, Isopropyl alcohol, gauge, # 40blade 1ml syringe,hypodermic needle- 25-30G Max- 0.1ml can beadministered

Must be done under anesthesia

3.Routes of drug administration in ratsd.Intraperitoneal injection(IP)

Requirements- Disposable gloves ,injection article,Isopropyl alcohol, gauge, 1-3 ml syringe, hypodermicneedle- 22-30G Max- 5ml can be administered

Insert needle into lower left/right quadrant of abdomen at a 30-degree angle.

3.Routes of drug administration in ratsf.Intravenous injection(IV)

Requirements- Restrainer, disposable gloves ,injectionarticle, Isopropyl alcohol, gauge,1ml syringe, hypodermicneedle- 25-30G Max- 0.5ml

Injection site- lateral vein

g. Oral gavage route in rats

Measure the distance from the tip of nose to the first rib. This is the length of needle that should be used

3.Sex determination in rats

Sexing is based upon anogenital distance. Maleshave a greater distance between the anus andurogenital opening.

4.Anesthesia techniques in rats

a. Induction of anesthesia by injectable anesthetics

Ketamine Ketamine+ Xylazine

Ketamine + Medetomidine

Duration 15-20min 30min 30-40min

Rat 75-100mg/kg IP 60mg/ kg + 5-10mg/kg, IP

75mg/ kg + 0.5mg/kg, IP

b.Induction of anesthesia by inhalation anesthetics

Requirements- Measuring jar, cotton, bell jar, chloroform/anesthetic ether,

5.Blood withdrawal techniques in rats

Blood volume- 6% of the body weight For eg 200g rat-12ml

Weekly 1% of the body weight blood can be collectedFor eg 200g rat- 2ml blood can be withdrawn

a.Blood collection by lateral tail vein. 1ml syringe, 25-30G needle, restrainer. Anesthesia is not required

b.By central tail artery1ml syringe, 25-30G needle,restrainer. Anesthesia is required

c. Blood collection by dorsal pedal vein

Needle- 23G

d. Blood collection by retro orbital sinus

Should be performed under anesthesia

Training is required

e. Blood collection by cardiac puncture

Should be performed under anesthesia

Syringe -10-20ml needle-20-23G in Rat 8-9ml blood can be collected, training is required.

6.Euthanesia in rats

a.Physical methods-Exsaguination,decapitation(for stress analysis ,cervical

dislocation.

b.Inhalation of gases-Carbon monoxide, carbon dioxide,CO2 + chloroform/halothane.

c.Drug administration-Barbiturate over dose (IP),Ketamine over dose (IM/IP),Sodium

pentothol (IP).