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Edit Comment Close Premium member Presentation Transcript WELCOME: WELCOMEEXPERIMENTAL ANIMALS IN PHARMACOLOGY: EXPERIMENTAL ANIMALS IN PHARMACOLOGY Neena ThomasCONTENTS: CONTENTS INTRODUCTION EXPERIMENTAL ANIMALS HANDLING AND RESTRAINT DRUG ADMINISTRATION BLOOD COLLECTION CONCLUSION REFERENCESINTRODUCTION: INTRODUCTIONEXPERIMENTAL ANIMALS: EXPERIMENTAL ANIMALS Mice Rat Rabbit Guinea pig Hamster Dog Cat Monkey Frog BIOMETHODOLOGY OF MOUSE (mus musculus): BIOMETHODOLOGY OF MOUSE ( mus musculus) Order-RodentiaPowerPoint Presentation: GENERAL BIOLOGY Smaller docile Acute hearing, well developed sense of smell, poor vision & short generation interval circadian rhythm Swiss albino mouse - widely used strainBASIC 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 CPHYSIOLOGICAL 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) 7-9HANDLING & 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 mousePowerPoint 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 avoidedFOR 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 & fingersRESTRAINT 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 ventilationMOUSE RESTRAINER: MOUSE RESTRAINER Different types available Plexiglasses, plastic cones,etc.SEXING: SEXING distance from the anus & genital papilla is greater in malesEXPERIMENTAL 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-RodentiaPowerPoint Presentation: GENERAL BIOLOGY Albino rat - widely used 2 strains: wistar sprague dawleyBASIC 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 CPHYSIOLOGICAL 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) 6-7HANDLING & RESTRAINT: HANDLING & RESTRAINT Grasp the base of tail wit thumb & forefinger FOR HEAVY RATS FOR SMALL RATSFOR CLOSE TREATMENTS/EXAMINATIONS: FOR CLOSE TREATMENTS/EXAMINATIONS Grasp the whole body with palm over the back & restrain forelimbs. rear legs & tail held by other handPowerPoint Presentation: Hold the skin of the back in the palm of your handRAT RESTRAINER: RAT RESTRAINEREXPERIMENTAL 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- lagomorphaPowerPoint Presentation: Albino NewZealand white – most common Non albino breeds-black & white DutchBASIC 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 CPHYSIOLOGICAL 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) 4-8HANDLING & 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 bodyPowerPoint Presentation: rabbit restrained in a lab coat wrapped around the body, and folded under its abdomenRABBIT RESTRAINER: RABBIT RESTRAINEREXPERIMENTAL 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- RodentiaBASIC 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/minPHYSIOLOGICAL 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) 6-12PowerPoint 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 handPowerPoint Presentation: EXPERIMENTAL USES Evaluation of bronchodilators Used in immunology Study of local anesthetics Study of vitamin C metabolism Study of TB and INH toxicityBIOMETHODOLOGY OF HAMSTER: BIOMETHODOLOGY OF HAMSTERPowerPoint 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 2-3yrsEXPERIMENTAL USES: EXPERIMENTAL USES Research in immunology & virology. Research on diabetes Used in cytological investigations,genetics,tissue culture, radiation researchFROG: 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 circulationCAT: CATEXPERIMENTAL USES : EXPERIMENTAL USES To study physiology of circulatory and neuro muscular systems Study of drugs affecting BP Investigations of ganglionic blockersMONKEY: 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 & DRUGSDRUG 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 weightPowerPoint Presentation: MEASURE FROM TIP OF NOSE TO LAST RIB TO DETERMINE PROPER INSERTION DEPTHPowerPoint 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 mandiblePowerPoint 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 possibilitiesPARENTERAL 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 bulbPowerPoint 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 veinPowerPoint 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 vasodilatorPowerPoint 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 veinPowerPoint 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 veinPowerPoint Presentation: GUINEA PIG Equipment : 27-30G,1inch needle Volume : 0.5ml Sites : lateral saphenous, cephalic vein, marginal ear veinINTRAPERITONEAL(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 vesselMOUSE: MOUSE Equipment : 23-27G,1/2 to 1-inch needle Volume : not to exceed 2ml Site : lateral aspect of lower left quadrant of abdomenRAT: RAT Equipment : 20-25G,1-1.5 inch needle Volume : not to exceed 10ml Site : lower quadrant of abdomenPowerPoint 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 anesthesiaSUBCUTANEOUS(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 MOUSEPowerPoint Presentation: RAT Equipment : 20-25G needle, syringe Volume : not to exceed 10ml Site : loose skin between shoulder bladePowerPoint Presentation: injected in the scruff of the neck with a minimum of immobilizationPowerPoint Presentation: RABBIT Equipment : 20-23G,1 inch needle, syringe Volume : 30-50 ml Sites : between the scapulaePowerPoint 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 skinPowerPoint Presentation: GUINEA PIG E quipment : 20-22G,3/4-1 inch needle, Short , heavy gauge needlesINTRAMUSCULAR(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 limitedPowerPoint Presentation: RAT Equipment : <21G , ½ inch needle & syringe Volume : not to exceed 0.3ml Sites : back &hind leg musclesPowerPoint 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 RABBITPowerPoint Presentation: GUINEA PIG Equipment : 20-22G , 1 inch needle, syringe Volume : 0.3ml Sites : lower back (lumbar) muscles & caudal muscles of thigh & quadricepsBLOOD 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 samplingPowerPoint 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/kgPowerPoint 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 vesselTools 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 collectionPowerPoint Presentation: Place mouse on a cage lid and grasp the loose skin behind the ears by the thumb and forefingerPowerPoint Presentation: Push the mouse into the restrainerPowerPoint Presentation: Leave the tail of the mouse outside the cover of the restrainerPowerPoint Presentation: Amputate the tip of the mouse tail by scissorsPowerPoint Presentation: Massage the tail and collect blood by pipettemanPowerPoint Presentation: Equipment restraining tube, scalpel blade, 27-gauge syringe needle, blood collection tube and cotton wool . Blood Collection From Saphenous VeinPowerPoint 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 appearPowerPoint Presentation: Blood collection blood collected in a sampling tube or microhaematocrit tube 100 microliters can be collectedPowerPoint 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 recoveredPowerPoint 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 anesthesiaPowerPoint Presentation: Use a sharp end glass capillary tube to penetrate the orbital conjunctiva and rupture the orbital sinusPowerPoint Presentation: Collect blood with a vialPowerPoint Presentation: AXILLARY BLEED Equipment : scalpel blade,3-5cc syringe Blood recovered from axillary region in a terminal exsanguination Requires anesthesia 1-2ml of blood recoveredPowerPoint 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 micePowerPoint Presentation: RABBIT MARGINAL EAR VEIN: Equipment : butterfly set Small vol of blood collected Sedation & vasodilation with acepromazinePowerPoint Presentation: CENTRAL EAR ARTERY Equipment : plain 20G, 1 inch needle attached to silicone coated tube Volume : 10ml/kg body wt Sedation & vasodilatation with acepromazinePowerPoint Presentation: CARDIAC PUNCTURE Equipment : 18G, 1.5 inch needle attached to vacuum tube Terminal blood collection Requires anesthesiaPowerPoint 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 methodPowerPoint 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 anesthesiaCONCLUSION: CONCLUSIONREFERENCES : 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.comPowerPoint Presentation: Thank you………… You do not have the permission to view this presentation. In order to view it, please contact the author of the presentation.