Common Laboratory Animal and Handling Techniques

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This is also a good complement to your "experimental animals used in pharmacology" presentation. Is it possible to have a copy? Thanks

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WELCOME:

WELCOME

Common Laboratory Animals and Handling Techniques:

Common Laboratory Animals and Handling Techniques Anuja.M.K. M pharm part 1 Pharmacology

Contents:

Contents Introduction Types of experimental animals used Animal handling Routes of drug administration in experimental animals Blood collection from experimental animals Conclusion References

Introduction:

Introduction Selection of an animal model is one of the most important step in any of the experimental pharmacological studies. Animal model preferred for the study must be producing similar disease profile as in the human.

Animal model selected should follow three main objectives::

Animal model selected should follow three main objectives : Use of an animal phylogenetically closer to man. Use of an animal in which the process under investigation is as close as possible to that in man. The Anatomy, Physiology and Biochemistry are considered to be similar .

Classification of experimental animals:

Classification of experimental animals

MOUSE (Mus musculus):

MOUSE ( Mus musculus ) Smallest laboratory animal Easy to keep, handle and require small place for housing, uniformly breed Common strain Swiss albino mice Sensitive to small doses of drugs

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Mice are only known species in which it is possible to grow totipotent embryonic stem (ES) cells in vitro can form germ line once re-injecting into a developing embryo. Mice use their tail to help in thermoregulation Used for bioassay of insulin and screening of analgesics by chemically induced writhing method.

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Used in the testing of teratogenicity. It provides good model for research on human diseases likes cancer, diabetes, ageing, atherosclerosis, immunological disease, autoimmune disorders, neurological dysfunction and other endocrine diseases .

RAT (Rattus norvegicus):

RAT (Rattus norvegicus) Rat is commonly used for the assay of different hormones and for the study of the oestrous cycle, mating behavior and fertility.

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Two inbred original strains of albino rats used Wistar rat: wide head and the ear is long where as tail length is less than body length Sprague Dawley rat: longer and narrow head, tail is longer than the body length

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Do not vomit (due to strong sphincter between stomach and esophagus and lack vomiting centre) Do not have tonsil and gallbladder Diffuse pancreas, so not a good model for type I diabetic studies Coprophagy (eating their own stool) Tail help in thermoregulation

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Used in research of behavior, pharmacology, physiology, neurosciences, immunogenetics, transplantation, cancer risk assessment, cardiovascular diseases and aging Various isolated tissues used are rat uterus, ileum, fundus strip, vas deference and anococcygeus muscle.

Guinea pig (Cavia porcellus):

Guinea pig (Cavia porcellus) Herbivorous and eats green foods seed and roots. Guinea pig are not able to synthesize required daily vit C Highly sensitive to histamine

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Highly sensitive to penicillin Serum contains an enzyme asparaginase, which shows antileukemic action Very susceptible to tuberculosis and anaphylactic shock Various isolated tissues used are guinea pig ileum tracheal chain, vas deferens and uterus

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Terminal portion of ileum used for screening of spasmodic and antispasmodic agents Ideal model for enteric ameobiasis, hypersensitivity, immune response anaphylactic shock, encephalomyelitis, tuberculosis and ascorbic acid metabolism

Gerbil(Meriones unguiculatus):

Gerbil(Meriones unguiculatus) Have length between rat and mice Also known as “sand rat” or “jirds”

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Ease in handling, mild and quiet nature Used as research animal in stroke, epilepsy, auditory studies (hearing curve similar to human) parasite and bacterial infections, lipid metabolism, heart disease studies

Hamster (Mesocricetus auratus):

Hamster (Mesocricetus auratus) Third commonly used laboratory animal Two species are commonly used -Golden or Syrian Hamster -Chinese Hamster

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Hamsters are used extensively in onco virus, influenza virus, respiratory syncitical virus (RSV) studies and vaccine production Cheek pouches do not have lymphatic drainage and hence they are ideal site for tissue transplants such as tumors and crafts European hamster is a more suitable model for highly concentrated and prolonged smoke inhalation studies

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Chinese hamster is useful for cytogenesis research. Syrian hamster is most commonly used in biomedical research because of availability and ease of production. Armenian hamster is more specific for the research to mutagenic and carcinogenic agents and for studying meiosis due to its susceptibility.

RABBIT(Oryctolagus cuniculus):

RABBIT(Oryctolagus cuniculus) Very docile animals Used for testing of large volume parenterals and for the screening and bioassay of insulin, antidiabetic and curaremimetic drugs

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New Zealand white rabbits have been used in the screening of different drugs for diseases like diabetes, diphtheria, tuberculosis, cancer and heart diseases. Employed in screening of antifertility drugs and for teratogenic studies Skin is sensitive to irritation. Hence used for irritancy tests Good model for the production of antibodies and antiserums

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Isolated heart, duodenum and ileum preparations are used for testing of drugs To test toxic effects of cosmetics and pharmaceuticals It has very simple cardiac conductive tissue, free of connective tissue and is an animal of choice for many cardiac studies. It is very sensitive to histamine. It cannot vomit

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It is an ideal animal for pharmacokinetic studies. Enzyme atropinesterase present in blood which degrade atropine. It has an ability to taste water, a characteristic absent in humans or rats. It is the only known mammal from which tubules of the kidney can be dissected with basement membrane intact.

Monkey (Macaca mulatta):

Monkey (Macaca mulatta) Structurally and functionally similar to man Suitable for undertaking psychopharmacological studies Uterus resembles humans and exhibiting regular menstrual periods

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It is an ideal model for pharmacokinetic study. Best for studying drugs acting on CNS, CVS, GIT and fertility. Require regular check up for rabies, tuberculosis and timely immunization. Used as primate model to study drug metabolism

Dog (Canis familiaris):

Dog (Canis familiaris) Most preferred large experimental animal Used for studying various anti arrhythmic, cardiovascular and autonomic drugs The advantages are small alimentary tract and easily get trained.

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Mongrel and beagles are the most preferred for the experiment purpose due to manageable size, moderate length of hair coat, docile nature and ease to handle. Stomach and intestinal tract resemble human It is also a good model for diabetes mellitus, reproduction, ulcerative colitis, open heart surgery, organ transplantation, central nervous system, safety pharmacology and toxicology

Cat (Felis catus):

Cat (Felis catus) Used in cardio vascular behavioral and biomedical research Has distinct nictitating membrane , used in screening of ganglionic drugs

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Cats are useful models in studying the transmission of vitamins and minerals to the foetus and newborn It is also used in the neuropharmacology, toxicology, oncology and chromosomal abnormality studies

Pig (Sus scrofa domestica):

Pig (Sus scrofa domestica)

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Preferred in skin related studies due to less hairy skin. Susceptible to bronchitis or pneumonia. Alimentary tract resembles human. Important animal model in cardiovascular research such as atherosclerosis, myocardial infarction etc..

Zebra Fish (Danio rerio):

Zebra Fish (Danio rerio) It is a suitable model for studying vertebrate embryo development and genetic techniques. Different stages of development can be observed under microscope

Frog (Rana tigrina):

Frog (Rana tigrina)

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Commonly used amphibian in pharmacology laboratory Oxygen can pass through their highly permeable skin and hence “breathe” largely through their skin. Camouflage is a common defensive mechanism in frogs. Anatomically their heart contains three chambers which is different from the other mammals used in the experiment

Chicken (Gallus domesticus):

Chicken (Gallus domesticus)

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Widely used due to ease of availability Good model in toxicology and pharmacology behavior studies Only model for studying Avian diseases Important models developed in chicken are chick comb method, chicken blood pressure measurement, heart rate, EEG, etc…

PIGEON (Columbia livia):

PIGEON (Columbia livia) Good screening model for antiemetic Used for standardization of cardiac glycosides Bioassay of prolactin Used for screening of intravenous anesthetics

ANIMAL HANDLING:

ANIMAL HANDLING Before restrain, first pet or sooth the animal by slow deliberate movements in the body Overcrowding near the animal cage should be avoided Noise should be kept as minimum Don’t hold animal too hard Never agitate the animal, it may become violent for self protection

MOUSE:

MOUSE One can handle it with the help of blunt forceps by grasping the skin behind the neck or body. (to transfer from one cage to another) Grasp the base of the tail with one hand and with the other grasp the loose skin behind its neck Hold the complete body by grabbing back of neck by using all fingers

RAT:

RAT Lift rat out of the cage by grasping the base of the tail and place on a soft surface Place your index and middle fingers alongside the rat’s head and your thumb and ring fingers under its forelegs. Use your index and middle fingers to secure its head and remaining fingers to support the body Hold the complete body by grabbing the back using complete palm

GUINEA PIG:

GUINEA PIG By using both hands, calmly grasp it with one hand under the chest and use your other hand to support its hindquarters Handle guinea pig with one hand, by holding its hindquarter

HAMSTER :

HAMSTER Hold the complete body by grabbing at the nap of the neck by thumb and index finger and grasp the complete body by using rest of fingers Hold the complete body by grabbing back by using complete palm

RABBIT :

RABBIT By using single hand, hold the pelvic region. This is to transfer rabbits from one cage to another By using both the hands, hold the complete hindquarter. By using both hands, calmly grasp it with one hand supporting back of neck and the other hand supporting its hindquarters

ROUTES OF ADMINISTRATION:

ROUTES OF ADMINISTRATION Feeding or oral gavage

Feeding or oral gavages:

Feeding or oral gavages Hold the rodent in hand carefully Measure the tube length from nose to the last rib of the rodent and mark it Give a gentle tight grip at back of the neck, so that it opens its mouth widely. Push the rodent head slightly upward and back to straighten esophagus and then either from right or left side of the teeth, insert the tube by gentle rotation to avoid the resistance

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5. Slowly pass the tube observing for the swallowing reflex and when desired length of the tube has been inserted, inject solution with the help of the syringe

INTRAPERITONEAL :

INTRAPERITONEAL

INTRAVENOUS :

INTRAVENOUS

INTRAMUSCULAR :

INTRAMUSCULAR

SUBCUTANEOUS :

SUBCUTANEOUS

INTRACARDIAC INJECTION:

INTRACARDIAC INJECTION

BLOOD COLLECTION FROM EXPERIMENTAL ANIMALS:

BLOOD COLLECTION FROM EXPERIMENTAL ANIMALS

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Terminal blood collection --whole blood withdrawal Non terminal blood collection --Single blood removal -- Multiple blood withdrawals

TERMINAL BLOOD COLLECTION:

TERMINAL BLOOD COLLECTION Done either after exsanguination of the animal by physical stunning or after submitting the animal to general anesthesia The blood from such animals can be collected by

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Withdrawal of blood from venacava or aorta after performing laparatomy Animals are subjected to exanguination after decapitation, the jugular vein or carotid artery is exposed given an incision and blood is directly collected using a syringe Blood can also be collected by retro-orbital bleeding of smaller animal like mice rats and hamsters.

NON TERMINAL COLLECTION OF BLOOD:

NON TERMINAL COLLECTION OF BLOOD Required for single or multiple withdrawals. Different peripheral veins of different animals may be used for collection of blood For multiple withdrawals, the amount of blood sample should not exceed 1% of total blood volume every 24 hours

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For single withdrawal of blood, upto 15% of total blood volume does not adversely effect animal. Withdrawal of more than 15%may lead to cardiac failure. A single withdrawal of upto 15%of total blood volume can be repeated after 3-4 weeks from the normal healthy animal.

TECHNICAL ASPECTS OF BLOOD REMOVAL:

TECHNICAL ASPECTS OF BLOOD REMOVAL Blood from tip of the tail Blood from superficial veins Permanent venous cannulation Retro-orbital bleeding Cardiac puncture

BLOOD COLLECTION FROM TIP OF TAIL:

BLOOD COLLECTION FROM TIP OF TAIL Common method used in rats and mice for collecting 0.1ml from the capillary blood is to cut sharply the tip of tail Adequate for multiple collection of blood samples for the determination of biochemical parameters like blood glucose, radio labelled drugs etc.. In tail less animals cardiac puncture is used for collection of blood

BLOOD FROM SUPERFICIAL VEINS:

BLOOD FROM SUPERFICIAL VEINS In larger animals superficial veins are used for blood collection The bore of the needle should as large as possible for ensuring rapid withdrawal of blood A check is made afterwards to ensure that no after bleeding occurs from this point

PERMANENT VENOUS CANNULATION:

PERMANENT VENOUS CANNULATION Used for chronic experiments which necessitate multiple blood collection at regular intervals The catheters are made to exit at the back of animal for only 2cms and capped with steel needles and at the time of blood collection a longer catheter is attached Thrombi formation due to clotting can be prevented by repeatedly filling the catheter with saline containing heparin

RETRO-ORBITAL BLEEDING:

RETRO-ORBITAL BLEEDING This technique for collection of blood should be used as the last resort ,by a well trained staff and be confirmed only to one of the eyes Employed in small animals like rat and mice when larger amount of blood is required which cannot be collected from tail vein or in tail less animals Should ideally be performed in anaesthetized animals

CARDIAC PUNCTURE:

CARDIAC PUNCTURE Generally employed in guinea pig, hamsters and gerbils. It is ideal to perform cardiac puncture in anaesthetized animal in order to reduce stress and facilitate smooth handling of animals Premedication with atropine helps to prevent cardiac arrhythmias

CONCLUSION :

CONCLUSION Different kinds of experimental animals are used as models for screening of various drugs. Pharmacologist should have a thorough knowledge about the animal handling, route of administration and techniques of blood collection

REFERENCES:

REFERENCES Drug discovery and evaluation pharmacological assays by H.Gerhard Vogels, page no:1371-1374 Screening methods in pharmacology, N.S.Parmar,Shiv Prakash,page no:45-53 Practical manual of Experimental and Clinical Pharmacology, Bikesh Medhi and Ajay Prakash,page no:6-35 www.google.com

Thank you:

Thank you