experimental animals used in pharmacology

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Published on June 14, 2012

Author: mk.anuja

Source: authorstream.com

WELCOME: WELCOME EXPERIMENTAL ANIMALS IN PHARMACOLOGY: EXPERIMENTAL ANIMALS IN PHARMACOLOGY Neena Thomas CONTENTS: CONTENTS INTRODUCTION EXPERIMENTAL ANIMALS HANDLING AND RESTRAINT DRUG ADMINISTRATION BLOOD COLLECTION CONCLUSION REFERENCES INTRODUCTION: INTRODUCTION EXPERIMENTAL ANIMALS: EXPERIMENTAL ANIMALS Mice Rat Rabbit Guinea pig Hamster Dog Cat Monkey Frog BIOMETHODOLOGY OF MOUSE (mus musculus): BIOMETHODOLOGY OF MOUSE ( mus musculus) Order-Rodentia PowerPoint Presentation: GENERAL BIOLOGY Smaller docile Acute hearing, well developed sense of smell, poor vision & short generation interval circadian rhythm Swiss albino mouse - widely used strain BASIC BIOLOGIC DATA: BASIC BIOLOGIC DATA ADULT BODY WEIGHT:MALE 20-40 gm ADULTBODY WEIGHT:FEMALE 18-35 gm LIFE SPAN 1.5-3 yrs FOOD CONSUMPTION 15gm/100gm/day WATER CONSUMPTION 15ml/100gm/day BREEDING ONSET:MALE 50days BREEDING ONSET:FEMALE 50-60days GESTATION PERIOD 19-21days BODY TEMPERATURE 36-37 0 C PHYSIOLOGICAL DATA: PHYSIOLOGICAL DATA HEART RATE (beats/min) 330-780 RESPIRATORY RATE per min 84-230 TIDAL VOLUME (ml) 0.1-0.23 Hb(g/100ml) 10-19 RBC(million/mm 3 ) 4.9-12.5 WBC(thousands/mm 3 ) 4-12 BLOOD VOLUME(%B.W) 7-9 HANDLING & RESTRAINT: HANDLING & RESTRAINT GENERAL PRINCIPLES: Use of proper restraint & handling techniques Acclimation period of upto 1 week Handle animals gently & firmly Use restraint devices when appropriate Chemical restraint for prolonged or potentially painful procedures MICE: MICE Grasp the middle of the tail between the thumb & index finger,& lift the mouse PowerPoint Presentation: HOW TO PICK UP A MOUSE WITH THUMP FORCEPS Forceps tip covered with rubber/polyethylene tubing Grasp the middle of the tail between the forceps Too much pressure avoided FOR TREATMENTS & CLOSE EXAMINATIONS : FOR TREATMENTS & CLOSE EXAMINATIONS held by tail & placed on a rough surface Grasp the loose skin over neck & shoulder with the thump & fingers RESTRAINT DEVICES: RESTRAINT DEVICES adapted to the restraint devices regularly monitored access to food or water released from restraint devices at least daily easy to clean & provide adequate ventilation MOUSE RESTRAINER: MOUSE RESTRAINER Different types available Plexiglasses, plastic cones,etc. SEXING: SEXING distance from the anus & genital papilla is greater in males EXPERIMENTAL USES: EXPERIMENTAL USES Acute toxicity studies Assay of insulin and analgesics in genetics and cancer research Testing of drug for teratogenicity BIOMETHODOLOGY OF RAT (rattus norvegicus) order-Rodentia: BIOMETHODOLOGY OF RAT (rattus norvegicus ) order-Rodentia PowerPoint Presentation: GENERAL BIOLOGY Albino rat - widely used 2 strains: wistar sprague dawley BASIC BIOLOGIC DATA: BASIC BIOLOGIC DATA ADULT BODY WEIGHT:MALE 300-500 gm ADULT BODY WEIGHT:FEMALE 250-300gm LIFE SPAN 2-3.5 yrs FOOD CONSUMPTION 10gm/100gm/day WATER CONSUMPTION 10-12ml/100gm/day BREEDING ONSET:MALE 65-110days BREEDING ONSET:FEMALE 65-110days GESTATION PERIOD 21-23days BODY TEMPERATURE 38-39 0 C PHYSIOLOGICAL DATA: PHYSIOLOGICAL DATA HEART RATE (beats/min) 300-500 RESPIRATORY RATE per min 65-180 TIDAL VOLUME (ml) 1.5 Hb(g/100ml) 12-17.5 RBC(million/mm 3 ) 7.2-9.6 WBC(thousands/mm 3 ) 6-12 BLOOD VOLUME(%B.W) 6-7 HANDLING & RESTRAINT: HANDLING & RESTRAINT Grasp the base of tail wit thumb & forefinger FOR HEAVY RATS FOR SMALL RATS FOR CLOSE TREATMENTS/EXAMINATIONS: FOR CLOSE TREATMENTS/EXAMINATIONS Grasp the whole body with palm over the back & restrain forelimbs. rear legs & tail held by other hand PowerPoint Presentation: Hold the skin of the back in the palm of your hand RAT RESTRAINER: RAT RESTRAINER EXPERIMENTAL USES: EXPERIMENTAL USES Testing of psycho-pharmacological agents Study of drugs on BP Gastric secretion studies Study of analgesics Toxicity studies Testing of teratogenicity and carcinogenicity BIOMETHODOLOGY OF RABBIT (oryctolagus cuniculus) order- lagomorpha: BIOMETHODOLOGY OF RABBIT ( oryctolagus cuniculus ) order- lagomorpha PowerPoint Presentation: Albino NewZealand white – most common Non albino breeds-black & white Dutch BASIC BIOLOGIC DATA: BASIC BIOLOGIC DATA ADULT BODY WEIGHT:MALE 2-5Kg ADULT BODY WEIGHT:FEMALE 2-6Kg GESTATION PERIOD 31 days LIFE SPAN 5-6 upto15yrs FOOD CONSUMPTION 5gm/100gm/day WATER CONSUMPTION 5-10ml/100gm/day RECTAL TEMPERATURE 38.5-39.5 0 C PHYSIOLOGICAL DATA: PHYSIOLOGICAL DATA HEART RATE (beats/min) 130-300 RESPIRATORY RATE per min 38-60 TIDAL VOLUME (ml) 19-24 Hb(g/100ml) 8-15 RBC(million/mm 3 ) 4.5-7 WBC(thousands/mm 3 ) 6-13 BLOOD VOLUME(%B.W) 4-8 HANDLING & RESTRAINT : HANDLING & RESTRAINT Never lift by ears While lifting, grasp the loose skin over the neck & shoulders with head directed away from holder. support the lower part of body PowerPoint Presentation: rabbit restrained in a lab coat wrapped around the body, and folded under its abdomen RABBIT RESTRAINER: RABBIT RESTRAINER EXPERIMENTAL USES: EXPERIMENTAL USES For pyrogen testing Study of agents affecting capillary permeability Study of antidiabetics and sex hormones Used for experimentation by dermal route Study of teratogens and embryotoxins For research on reproduction BIOMETHODOLOGY OF GUINEA PIG (Cavia porcellus) order- Rodentia: BIOMETHODOLOGY OF GUINEA PIG (Cavia porcellus) order- Rodentia BASIC BIOLOGIC DATA: BASIC BIOLOGIC DATA ADULT BODY WEIGHT:MALE 800-200gm ADULT BODY WEIGHT:FEMALE 250-320gm GESTATION PERIOD 65-72days LIFE SPAN 4-5yrs FOOD CONSUMPTION 6gm/100gm/day WATER CONSUMPTION 10ml/100gm/day HEART RATE 230-380beats/min RESPIRATORY RATE 42-104/min PHYSIOLOGICAL DATA: PHYSIOLOGICAL DATA HEART RATE (beats/min) 260-400 RESPIRATORY RATE per min 70-104 TIDAL VOLUME (ml) 1.8 Hb(g/100ml) 8-15 RBC(million/mm 3 ) 5-6 WBC(thousands/mm 3 ) 4-11 BLOOD VOLUME(%B.W) 6-12 PowerPoint Presentation: how to hold a guinea pig for treatment HANDLING & RESTRAINT Place the thumb beneath the jaw of guinea pig & support the hindquarter with other hand PowerPoint Presentation: EXPERIMENTAL USES Evaluation of bronchodilators Used in immunology Study of local anesthetics Study of vitamin C metabolism Study of TB and INH toxicity BIOMETHODOLOGY OF HAMSTER: BIOMETHODOLOGY OF HAMSTER PowerPoint Presentation: 2 SPECIES syrian/golden (mesocricetus auratus) chinese (cricetulus griseus) BASIC BIOLOGIC DATA: BASIC BIOLOGIC DATA ADULT BODY WEIGHT: Syrian 80-90g ADULT BODY WEIGHT: Chinese 35-40g LIFE SPAN 2-3yrs EXPERIMENTAL USES: EXPERIMENTAL USES Research in immunology & virology. Research on diabetes Used in cytological investigations,genetics,tissue culture, radiation research FROG: FROG (Rana tigrina) EXPERIMENTAL USES: EXPERIMENTAL USES Study of action of drugs on CNS, heart, neuromuscular junction To determine retinal toxicity of drugs. DOG (Canis familaris): DOG ( Canis familaris) EXPERIMENTAL USES : EXPERIMENTAL USES Study of drugs on BP For classical works on diabetes & study of hypoglycemic agents. Study of gastric secretions Conducting experiments on circulation CAT: CAT EXPERIMENTAL USES : EXPERIMENTAL USES To study physiology of circulatory and neuro muscular systems Study of drugs affecting BP Investigations of ganglionic blockers MONKEY: MONKEY (Macaca mulatta) EXPERIMENTAL USES : EXPERIMENTAL USES in the field of virology, parasitology, immunology and in immunosuppression, nutrition, reproduction etc. BLOOD COLLECTION & ADMINISTRATION OF FLUIDS & DRUGS: BLOOD COLLECTION & ADMINISTRATION OF FLUIDS & DRUGS DRUG ADMINISTRATION TECHNIQUES: DRUG ADMINISTRATION TECHNIQUES Route - nature of the agent ,animal & the purpose of the administration Oral (p.o) - by addition to the food/water Gavage needles : Through mouth into stomach or lower esophagus Exact amount administered Amount limited to 1% of body weight PowerPoint Presentation: MEASURE FROM TIP OF NOSE TO LAST RIB TO DETERMINE PROPER INSERTION DEPTH PowerPoint Presentation: Mice, rat - doesn’t require anesthesia Rabbit - restrainer used. wooden or plastic dowel with a hole in the center is held crossways behind the incisors Guinea pig -sedatives administered feeding needle is introduced in the space between left incisors & molars & gently direct it caudally towards the right ramus of the mandible PowerPoint Presentation: C omplications associated with gastric intubation : damage to esophagus administration of substance into trachea . Careful and gentle passage of the feeding needle will greatly reduce these possibilities PARENTERAL ADMINISTRATION: PARENTERAL ADMINISTRATION INTRAVENOUS (iv): MOUSE Equipment : 27-30G needle,1ml tuberculin syringe, mouse holder, warming lamp Volume : not to exceed 0.2ml Site : Lateral tail vein best results obtained if tail is immersed in warm water for 5- 10sec or the mouse warmed for 5-15min in the cage with a warming lamp with a 40-100W bulb PowerPoint Presentation: RAT Equipment : 20-25G needle,1-3ml syringe, rat holder, warming lamp Volume : not to exceed 0.5ml Major site : lateral tail veins Other sites : lateral saphenous vein, sublingual vein,dosal penile vein PowerPoint Presentation: RABBIT Equipment : 20-25G needle with syringe or butterfly infusion sets, rabbit holder Volume : not to exceed 5ml Sites : marginal ear vein Xylene/acepromazine as vasodilator PowerPoint Presentation: After shaving, the vein is easily visible . PowerPoint Presentation: The site should be disinfected before injection. PowerPoint Presentation: Stretch out the ear, supporting it with a finger underneath and insert the needle in the vein PowerPoint Presentation: The needle is visible when it is inserted in the vein. PowerPoint Presentation: Secure the needle in the vein by using your thumb. To ensure that the needle is in the vein, aspirate to get blood into the syringe . PowerPoint Presentation: After ensuring that the needle is properly placed, inject the fluid steadily into the vein PowerPoint Presentation: GUINEA PIG Equipment : 27-30G,1inch needle Volume : 0.5ml Sites : lateral saphenous, cephalic vein, marginal ear vein INTRAPERITONEAL(i.p): INTRAPERITONEAL (i.p) Most frequently used parenteral routes in rodents Lower left quadrant of abdomen Aspirate prior to injection Yellow fluid : needle could be in bladder Greenish fluid : needle could be in intestine Blood : may have entered blood vessel MOUSE: MOUSE Equipment : 23-27G,1/2 to 1-inch needle Volume : not to exceed 2ml Site : lateral aspect of lower left quadrant of abdomen RAT: RAT Equipment : 20-25G,1-1.5 inch needle Volume : not to exceed 10ml Site : lower quadrant of abdomen PowerPoint Presentation: RABBIT E quipment : 19-20G needle, syringe, restraining board Volume : 50-100ml Site : lower abdominal quadrant Chemical restraint with ketamine GUINEA PIG E quipment : 19-22G needle with syringe, restraining board Volume : 10-15ml Site : lower abdominal quadrant Light anesthesia SUBCUTANEOUS(sc): SUBCUTANEOUS (sc) Equipment : syringe & 25-27G, 1/2 to 3/4inch needle Volume : not to exceed 2-3ml Site : loose skin between the shoulder blades MOUSE PowerPoint Presentation: RAT Equipment : 20-25G needle, syringe Volume : not to exceed 10ml Site : loose skin between shoulder blade PowerPoint Presentation: injected in the scruff of the neck with a minimum of immobilization PowerPoint Presentation: RABBIT Equipment : 20-23G,1 inch needle, syringe Volume : 30-50 ml Sites : between the scapulae PowerPoint Presentation: To give the subcutaneous injection begin by lifting up the skin. PowerPoint Presentation: Choose a place where the skin is very loose, for example over the shoulder blades. PowerPoint Presentation: Aspirate before injecting in order to ensure that you have not entered a blood vessel. PowerPoint Presentation: If no blood is seen after aspirating, inject the fluid under the skin PowerPoint Presentation: GUINEA PIG E quipment : 20-22G,3/4-1 inch needle, Short , heavy gauge needles INTRAMUSCULAR(im): INTRAMUSCULAR (im) MOUSE Equipment : syringe & 26-30G, 1/2-inch needle Volume : not to exceed 0.05ml Sites : back &hind leg muscles Not recommended Volume injected limited PowerPoint Presentation: RAT Equipment : <21G , ½ inch needle & syringe Volume : not to exceed 0.3ml Sites : back &hind leg muscles PowerPoint Presentation: Equipment : 22-23G,1 inch needle, syringe Volume : 0.5ml Sites : back muscles lateral to the vertebrae & caudal to ribs or lateral thigh muscles RABBIT PowerPoint Presentation: GUINEA PIG Equipment : 20-22G , 1 inch needle, syringe Volume : 0.3ml Sites : lower back (lumbar) muscles & caudal muscles of thigh & quadriceps BLOOD COLLECTION TECHNIQUES: BLOOD COLLECTION TECHNIQUES 10-15% of total blood volume or 1% of body weight is the maximum amount of blood that should be collected at one time maximum amount of blood be withdrawn only once every two weeks .05% of B.W is the amount withdrawn in multiple sampling PowerPoint Presentation: Site preparation Area of injection cleaned with alcohol Some procedures requires anesthesia Veins dilation accomplished Total blood volume Mouse : 70-80ml/kg Rat : 50-65ml/kg Rabbit : 45-70ml/kg Guinea pig : 65-90ml/kg PowerPoint Presentation: MOUSE TAIL VEIN VENIPUNCTURE For collection of small amount of blood (0.1 ml ) Lateral or dorsal veins used Gentle aspiration to avoid collapse of vessel Tools for Blood Collection by amputation of tail tip : Tools for Blood Collection by amputation of tail tip alcohol cotton ball mouse restrainer Scissors Pipetteman and tips A vial for blood collection PowerPoint Presentation: Place mouse on a cage lid and grasp the loose skin behind the ears by the thumb and forefinger PowerPoint Presentation: Push the mouse into the restrainer PowerPoint Presentation: Leave the tail of the mouse outside the cover of the restrainer PowerPoint Presentation: Amputate the tip of the mouse tail by scissors PowerPoint Presentation: Massage the tail and collect blood by pipetteman PowerPoint Presentation: Equipment restraining tube, scalpel blade, 27-gauge syringe needle, blood collection tube and cotton wool . Blood Collection From Saphenous Vein PowerPoint Presentation: The mouse is restrained in a plastic tube with air holes in the end. PowerPoint Presentation: Shaving Avoid cutting the skin by holding the blade as flat as possible. PowerPoint Presentation: The vein is now easily visible. PowerPoint Presentation: vein punctured with a 27-gauge needle held at approximately 90 degrees to the skin. PowerPoint Presentation: Bleeding Immediately after puncture a drop of blood will appear PowerPoint Presentation: Blood collection blood collected in a sampling tube or microhaematocrit tube 100 microliters can be collected PowerPoint Presentation: TOE CLIPPING/ TAIL CLIPPING Unacceptable method CARDIAC PUNCTURE Equipment : 0.9-0.5mm needle Terminal blood collection Requires anesthesia Withdraw blood slowly Upto 1ml recovered PowerPoint Presentation: ORBITAL SINUS VENIPUNCTURE Equipment : capillary tubes Convenient & easy method Upto 0.5ml can be recovered Tube directed into orbital sinus Inserted into medial canthus of eye Requires anesthesia PowerPoint Presentation: Use a sharp end glass capillary tube to penetrate the orbital conjunctiva and rupture the orbital sinus PowerPoint Presentation: Collect blood with a vial PowerPoint Presentation: AXILLARY BLEED Equipment : scalpel blade,3-5cc syringe Blood recovered from axillary region in a terminal exsanguination Requires anesthesia 1-2ml of blood recovered PowerPoint Presentation: RAT Lateral tail vein venipuncture : method same as in mice Ventral tail artery Equipment : 22G needle,3cc syringe Anesthesia required Cardiac puncture : same as in mice Orbital sinus venipuncture : same as in mice PowerPoint Presentation: RABBIT MARGINAL EAR VEIN: Equipment : butterfly set Small vol of blood collected Sedation & vasodilation with acepromazine PowerPoint Presentation: CENTRAL EAR ARTERY Equipment : plain 20G, 1 inch needle attached to silicone coated tube Volume : 10ml/kg body wt Sedation & vasodilatation with acepromazine PowerPoint Presentation: CARDIAC PUNCTURE Equipment : 18G, 1.5 inch needle attached to vacuum tube Terminal blood collection Requires anesthesia PowerPoint Presentation: GUINEA PIG LATERAL SAPHENOUS VEIN VENIPUNCTURE Equipment : 22G needle/ scalpel blade Small vol blood collected LATERAL SAPHENOUS VEIN(VACUUM ASSISTED) Larger vol blood collected META TARSAL VEIN TOE CLIPPING Unacceptable method PowerPoint Presentation: CRANIAL VENACAVA Equipment : 22-23G needle,3-6 cc syringe Site : manubrium of sternum Repetitive collection of >1ml blood (upto 7ml/kg) Anesthesia required CARDIAC PUNCTURE Equipment : 20-22G needle,12-20cc syringe Not recommended as a repetitive blood sampling procedure Requires anesthesia CONCLUSION: CONCLUSION REFERENCES : REFERENCES Fundamentals of Experimental Pharmacology . M.N. Ghosh; 2 nd Edn: Pg :3-15. Handbook of Experimental Pharmacology . S.K.Kulkarni, 3 rd Edn; Pg: 12-14. Experimental Pharmacology, Volume 2 . J.K. Grover; Pg: 8-20. Drug discovery and evaluation . H.Gerhard Vogel, 2 nd Edn; Pg:1371-1378 www.google.com PowerPoint Presentation: Thank you…………

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