1. Pharmacology – I (Practical) [PHL 313]
Dr. Mohd Nazam Ansari

2. Lab – 1: Different Laboratory Animals and Their Application
PHARMACOLOGY is the branch of science which deals with study of drugs on living systems
EXPERIMENTAL PHARMACOLOGY: deals with study of effect of various Pharmacological agents on different animal species
AIMS :
To find out the therapeutic agent suitable for human use
To study the toxicity of the drugs
To study the mechanism and site of action of drugs
PHARMACOLOGY
Discovery of new drugs or to study the actions of existing drugs
Preclinical Clinical
Two ways Phase -1,2,3,4
Intact animal study – invivo
Isolated organ study – invitro

3. Clinical Trial Phase I: Phase II:
A small group (20-100) of healthy volunteers
The safety, tolerability, pharmacokinetics, and pharmacodynamics of a drug
These trials are often conducted in an inpatient clinic, where the subject can be observed by full-time staff
Pay ranges from a small amount of money for a short period of residence, to a larger amount of up to approx $6000 depending on length of participation.
Phase II:
Safety assessments in a larger group of healthy volunteers and patients (20-300)

4. Phase III:
Randomized controlled multicenter trials on large patient groups (300–3,000 or more depending upon the disease/medical condition studied)
Phase III trials are the most expensive, time- consuming and difficult to design and run, especially in therapies for chronic medical conditions
Phase IV:
Is also known as Post Marketing Surveillance Trial.
Harmful effects discovered by Phase IV trials may result in a drug being no longer sold, or restricted to certain uses

5. Experimental pharmacology – great importance
Experimenter should take outmost care
Sacrificing – human
Euthanasia – painless killing
Laboratory animals – breaded and handled in laboratory
Rat
Mice
Guinea pig
Rabbits
Frogs
Other: Cat, Dog, Monkey, Pigeon etc.

6. RATS Species – Rattus norvegicus
Albino rats of Wistar strain are commonly used
Other strains –
Wistar kyoto rat
Sprague Dawley rat
Biobreeding (BBDP) rat
Long-Evans rat
Zucker rat Genetically modified rats
Hairless rats (Rowett nude, Fuzzy, Shorn)
RCS rats

7. Wistar rat Wistar rats are strain of albino rats
belonging to the species Rattus norvegicus.
This strain was developed at the Wistar Institute in for use in biological and medical research, and is notably the first rat strain developed to serve as a model organism at a time when laboratories primarily used Mus musculus, or the common House mouse.
The Wistar rat is currently one of the most popular strains used for laboratory research.
It is characterized by its wide head, long ears, and having a tail length that is always less than its body length.
The Sprague Dawley rat and Long-Evans rat strains were developed from Wistar rats. Wistar rats are more active than other strains like Sprague Dawley rats.

8. Sprague Dawley rat
The Sprague Dawley rat is multipurpose breed of albino rat used extensively in medical research.
Its main advantage is its calmness and ease of handling.
The adult body weight is 250–300g for females, and 450–520g for males. The typical life span is 2.5–3.5 years. These rats typically have increased tail to body length ratio compared with Wistar rats.

9. Biobreeding rat
Biobreeding Diabetes Prone rats (or BBDP rat) are inbred rat strain that spontaneously develops autoimmune Type 1 Diabetes.
BB rats are used as an animal model for Type 1 diabetes.
The strain re-capitulates many of the features of human type 1 diabetes, and has contributed greatly to the research of T1D pathogenesis

10. Long-Evans rat
Long-Evans rats are strain of rats belonging to the species Rattus norvegicus.
This strain was developed by Drs. Long and Evans in 1915 by crossing several Wistar females with a wild gray male.
Long Evans rats are white with a black hood, or occasionally white with a brown hood.
They are utilized as a multipurpose model organism, frequently in behavioral and obesity research.

11. Zucker rat
Zucker rats were bred to be a genetic model for research on obesity and hypertension.
There are two types of Zucker rat: a lean Zucker rat; and the characteristically obese (or fatty) Zucker rat, capable of weighing up to 1 kilogram (2.2 lb)—more than twice the average weight.
Obese Zucker rats have high levels of lipids and cholesterol in their blood, and gain weight from an increase in both the size and number of fat cells.
Obesity in Zucker rats is primarily linked to their hyperphagic nature, an excessive hunger.

12. Hairless rats
Hairless rats provide researchers with valuable data regarding compromised immune systems and genetic kidney diseases.
The more common ones are denoted as rnu (Rowett nude), fz (fuzzy), and shn (shorn).
Rowett nudes, first identified in 1953 in Scotland, have no thymus. The lack of this organ severely compromises their immune system, infections of the respiratory tract and eye increasing the most dramatically.
Fuzzy rats were identified in The leading cause of death among these rats is a progressive kidney failure.
Shorn rats were bred from Sprague Dawley rats in They also suffer from severe kidney problems.

13. RCS rats
The Royal College of Surgeons (RCS) rat is the first known animal with inherited retinal degeneration.

14. ADVANTAGES AND CHARACTERISTICS:
small in size
drug to be tested required in small quantity
vomiting center is absent – oral administration can be done
gall bladder and tonsils are absent
Continuous flow of bile into intestine, This facilitates the study of the drugs acting on bile, cholesterol reabsorption etc.
In stomach, fundus and pyloric parts have clear lining between them
Gastric acid secretion is continuous

15. EXPERIMENTAL USE: (Adult weight: 200-250g, Age: 1.5 months)
Psychopharmacological studies
Study of analgesics and anticonvulsants
Bioassay of various hormones, such as insulin, oxytocin, vasopressin
Isolated tissue preparations like uterus, stomach, vas deferens, aortic strip, heart etc.
Chronic study on blood pressure
Gastric acid secretion studies
Hepatotoxicity studies
Acute and chronic toxicity studies

16. MICE ( Mus musculus) Swiss albino mice are commonly used species
Other strains are – Balb/C and C-57
ADVANTAGES AND CHARACTERISTICS:
Smallest
Less drug required
Easy to handle
Cheap
EXPERIMENTAL USES: (Adult weight: 20-25g, Age: 1 month)
Toxicological studies specially acute and subacute toxicity
Bioassay of insulin
Screening of analgesic and anticonvulsant
Screening of chemotherapeutic agents
Studies related to genetics and cancer research
Drugs action on CNS

17. GUINEA PIGS ( Cavia porcellus )
ADVANTAGES AND CHARACTERISTICS :
Docile animals
Highly susceptible to TB and anaphylaxis
Highly sensitive to histamine, penicillin
Required exogenous vitamin C in diet
EXPERIMENTAL USES (Adult weight: g, Age: 3 months)
Evaluation of bronchodilators
Study of histamine and antihistamines
Bioassay of digitalis
Evaluation of local anesthetics
Hearing experiments because of sensitive cochlea
Isolated tissues especially ileum, tracheal chain, heart etc.
Study on TB and ascorbic acid metabolism

18. RABBITS (Lupas cuniculus)
ADVANTAGES AND CHARACTERISTICS :
Docile animal with large ears
New Zealand white strains are widely used
It has huge caceum and long appendix
Enzyme atropine esterase is present in rabbit liver and plasma so it can tolerate large doses of belladona (atropine)
EXPERIMENTAL USES (Adult weight: Kg, Age: 5-6months)
Pyrogen testing
Bioassay of anti-diabetic, and sex hormones
Drugs used in glaucoma
Pharmacokinetic studies
Studies related to antifertility agents
Isolated preparations like heart, duodenum, ileum, etc.
Study on local anaestetics
Study on miotic and mydratics

19. FROGS (Rana tigrina) ADVANTAGES AND CHARACTERISTICS :
Used before 200 years
Easily available during rainy season
Amphibian animal and safe to handle
Cannot breed in lab
EXPERIMENTAL USES
Isolated preparations, rectus abdominis muscle & heart
Drugs acting on CNS
Drugs acting on NMJ

20. Housing & Maintenance of Laboratory animals
Institutional or departmental ethical committee
Animal House
Separate building at a quiet atmosphere, undisturbed by traffic
Should be hygienic & protected from extremes of climate
Good ventilation
Exhaust fan
Roof should be 10 feet high
Large no. of small rooms
Quarantine area
Extra space for office, surgery, washing and sterilizing, kitchen & incinerators
No overcrowding permitted
Animal Cages:
Plastic, galvanised iron, aluminium etc.
Anodised aluminium is best
Size
Tray for food and water bottle
Bedding: rice husk or paper
Proper labeling

21. Clean drinking water Ventilation Food Free movement Resting
Following requirements should be stressed on mind:
Clean drinking water
Ventilation
Food
Free movement
Resting
Temperature
Humidity
Diseases
Handling
Mental Health

22. Housing and Routine Care: Animal Crowding
Animal crowding in a cage affects environmental quality (the accumulation of urine, for example, leads to excess ammonia and moisture).
Crowding can also cause newborn pups to be injured or killed. Crowding is a special concern for multiple litters in a cage since pups grow very quickly and rapidly increase their output of excreta.
In particular, if the mother is about to give birth to a second litter, the first litter should be weaned and removed to new cages to prevent smothering and trauma of the newborns.

23. Routine Sanitation
Routine sanitation and environmental controls are necessary for protecting animal health and for minimizing the introduction for non-experimental variables which could undermine the quality of research data.
Sanitation schedules vary according to the type of mouse caging.
Based on the types of caging and bedding in use at your facility, your institution will have a standard operating procedure (SOP) on the sanitation schedule for mouse cages.