lab animals - kush

8/3/2019 Lab Animals - Kush

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Lab. animals

Presented by Dr. Kush Pathak 1


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Introduction

Definition

History

Reasons for use of animals

Animals used in research

Importance of animals in biomedical research

Animal Selection

Facility Preparation Animal care through experiment

Collection from Animals

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Content


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Research

Important bodies which enforce standards

Records

The Principles of Humane Experimental Technique

Conclusion

References

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Virtually every major medical advance of thelast 100 years (as well as advances in veterinary

medicine) has depended on research with animals.

Animal studies have provided the scientific

kno

wledge that allo

ws health care pro

viders to

improve the quality of life for humans and

animals by preventing and treating diseases and

disorders, and by easing pain and suffering.

Foundation for Biomedical Research,

Understanding the Use of Animals in Biomedical

Research

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INTRODUCTION

Animal testing, also known as animalexperimentation, animal research, and in

vivo testing, is the use of nonhuman animals inexperiments.

Number of vertebrate ranges from the tens ofmillions to more than 100 million used annually.

Number

of I

nvertebrates mice, rats, birds,fish, frogs. estimate of mice and rats used in

the United States alone in 2001 was 80 million.

Most animals are euthanized after being used in

an experiment.

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Sources of laboratory animals vary betweencountries and species; most animals are purpose

bred, while others are caught in the wild orsupplied by dealers.

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Definition

"vivisection" means the "cutting up" of a living

animal.

Animal testing, als

o

kno

wn as animalexperimentation, animal research, and in

vivo testing, is the use of nonhuman animals in

experiments.

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History

Earliest animal testing are found in the writingsof the Greeks in the 2nd and 4th centuries B.C.

Aristotle (384322 B.C) and Erasistratus (304258

B.C) were among the first to performexperiments on living animals.

Galen, a physician in 2nd-century Rome, dissected

pigs and goats, and is known as the "father of

vivisection.

Moorish Spain who also practiced dissection,introduced animal testing as an experimental

method of testing surgical procedures before8


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1946 Post WWII boom in public funding of science.

1950 Chicago Five Animal Care Panel

Louis Pasteur convincingly demonstrated

the germ theory of medicine by

inducing anthrax in sheep.

Ivan Pavlov famously used dogs to

describe classical conditioning. On November 3,

1957, a Russian dog, Laika, became the first ofmany animals to orbit the earth.

1963 Guide for Lab Animal Facilities and Care.

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1966 Lab Animal Welfare Act.

In the 1970s, antibiotic treatments and vaccines

for leprosy were developed using

armadillos, then given to humans.

1973 PHS Policy.

1985 PHS (Public Health Service Policy) Act.

1991 PHS Act Amended.

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Reasons for use of Animals in

researchOrgans and body systems similar to humans and other animals

1. Similar anatomies

2. Even though they may be different sizes and

shapes, humans and animals both have bonesthat contain marrow and produce blood cells.

3. Same hormones that aid digestion and regulate

the reproductive cycle. 11


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Susceptible to same diseases that affect humans

1. Cancers, diabetes, and heart diseases. Eg. dog

whose cardiovascular system closely resembles

that of the human.

2. Many successful medical breakthroughs that

can be attributed to the dog and that are now

commonly applied to humans includes Heart

transplantation, Coronary bypass surgery,

Artificial heart valves, Pacemaker implantation,Angioplasty

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Short life span allows animals to be studied throughout their

entire life

1. Rodents such as rats and mice

Environment easily controllable to keep experimental variablesto minimum

1. Under stress, the research is not reliable.

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Animals used in research

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Invertebrates

i. Although many more invertebrates than

vertebrates are used, these experiments arelargely unregulated by law.

ii. Most used invertebrate species are Drosophila

melanogaster, a fruit fly, and Caenorhabditiselegans, a nematode worm.

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C. elegans The worm's body is completely

transparent.

Studies in the fly D. melanogaster can use an

amazing array of genetic tools

Vertebrates

Mice

Rats

Hamsters

Dogs, cats, rabbits, farm animals, fish, frogs,

birds, nonhuman primates

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Animal Selection

Animal qualityI. Health quality

a. Conventional

b. Specific pathogen free (SPF)

c. Germ free

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Conventional - An animal is colonized bythe burden of resident microorganisms which is

normally ass

ociated with its particular species.

Germ-free animal 1. Animals that have no microorganisms living in or

on it.

2. Such animals are raised within germfree isolators

in order to control their exposure to viral,

bacterial or parasitic agents

3. When known strains of bacteria or micro biota are

introduced to a germ-free animal, it usually

referred to as a gnotobiotic animal.

4. Used in the study of probiotic research 19


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Specific pathogen free (SPF)

1. Animals that are guaranteedf

reeof

particular pathogens.

2. It is always accompanied by a list of the absent

pathogens.

3. Use of SPF animals ensures that specified

diseases do not interfere with an experiment

4. For example, absence of respiratory pathogens

such as influenza is desirable when investigating

a drug's effect on lung function.

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II. Genetic quality

a. Out bredb. Inbred

c. Hybrid

d. Mutant

e. Transgenic

f. Knock out

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Out bred animals

Animals that are generated from breeding two

genetically dissimilar strains of the same species.

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Wistar Rat Syrian HamsterSprague Dawley Rat

Dunkin Hartley

Guinea-pig

New Zealand White

Rabbit


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Inbred animals

Individuals of a particular species which are

nearly identical to each other in genotype due tolong inbreeding.

Frequently used in laboratories.

For some experiments, genetic diversity in the

test population may be desired. Thus out bred

strains of most laboratory animals are alsoavailable

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DBA/2J

C57Bl/6J

BALB/cA

C3H/Hen


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Hybrid

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BCB6F1 ( BALB/cA + C57BL/6J)


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Mutant

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Nude(BALB/c- nu )

Result from an instance ofmutation, which is a base-pair sequence change

within the DNA of a gene orchromosome, resulting in the creation of a new

character ortrait not found in the wild type.


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Transgenic

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Refers to an animal in which there has been a deliberate

modification of the genome (material responsible for inherited

characteristics) - in contrast to spontaneous mutation (FELASA, 1995 -

Federation for Laboratory Animal Science Associations)

Foreign DNA is introduced into the animal, using recombinant DNAtechnology.

Must be transmitted through the germ line so that every cell, including

germ cells, of the animal contain the same modified genetic material.


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Knockout animals

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129 iNOS(knockout gene)129 wild type

A knockout mouse is a genetically engineered mouse in which researchers haveinactivated, or "knocked out," an existing gene by replacing it or disrupting it with an

artificial piece ofDNA.

Causes changes in a mouse's phenotype, which includes appearance, behavior & other

observable physical and biochemical characteristics


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The importance of animals in

biomedical research

Allergies

Arthritis

Asthma

Birth defects Cancer

Tuberculosis

Epilepsy

Heart disease

kidney disease

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Lye disease

Ulcers

Measles Influenza

Hypertension

Glaucoma

Diabetes Bronchitis

Leukemia

Deafness

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HOUSING

It should

Provide adequate space & a comfortable environment.

Provide an escape proof enclosure that confines animals safely.

Provide easy access to food and water.

Provide adequate ventilation.

Keep the animals dry and clean.

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Should be constructed of sturdy, durable

materials and designed to minimize cross

infection between adjoining units.

Attention must be given to

eliminating sharp edges

and broken wires,

keeping cage in good

condition.

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Social environment

The social environment includes all interactions

among individuals of a group or among those able

to communicate.

Population density can affect reproduction,

metabolism, immune responses and behavior.

Should enrich the environment as appropriate to

the species, especially when animals will be held

for long periods.

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SPACE RECOMMENDATIONS FOR

LABORATORY ANIMALS

Animal Animal Weight (grams)Weight (grams) Floor area/animalFloor area/animal

(inches)(inches)

HeightHeight(inches)(inches)

MiceMice

RatsRats

25

500

6.06.0

8.08.012.012.0

15.015.0

17.017.0

23.023.0

29.029.0

40.040.0

60.060.0

70.070.0

55

5555

55

77

77

77

77

77

7735


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Animal Animal Weight (grams)Weight (grams) Floor area/animalFloor area/animal

(inch(inch22

))

HeightHeight

(inches)(inches)

HamstersHamsters

Guinea pigGuinea pig

100

350

>350

10.0

13.0

16.019.0

60.0

101.0

66

66

6666

77

77

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Animal Animal Weight (kg)Weight (kg) Floor area/animalFloor area/animal

(ft(ft22))

HeightHeight(inches)(inches)

RabbitsRabbits

CatsCats

DogsDogs

5.4

44

>4>4

30

1.51.5

3.03.0

4.04.0

5.05.0

3.03.0

4.04.0

8.08.0

12.112.1

24.024.0

1414

1414

1414

1414

2424

2424

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ACTIVITY

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Provision should be made for animals with specialized loco motor

pattern to express these patterns,

especially when the animals are

held forlong periods.

e.g. ropes, bars, and perches are

appropriate for branching non-

human primates.


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ANIMAL SPECIFICATION

Normal

Special a. Pregnancy,

b. Suckling,

c. Old,

d. Animal with some treatment

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Facility Preparation

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Animal room Research roomWash room


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Environment control

Temperature and humidity affect metabolism and behavior.

Air conditioning is an effective means of regulating these

environmentalparameters for laboratory animals.

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VENTILATION

The purpose of ventilation is to supply adequate oxygen;

Remove thermal loads caused by animal respiration,

Lights and equipment;

Dilute gaseous and particulate contaminants;


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16-20 fresh air changes/hourappears to provide adequate ventilation

for animal facilities.

Ventilation also provides an odor free environment.

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ILLUMINATION

Light can affect thephysiology andbehaviorof various animals.

Lighting should be uniformly diffused throughout animals facilities

and provide sufficient illumination.

Fluorescent tube lights fixed in such a way facilitate uniform

distribution of light.

Light intensity of350-400 lux one metre above the floor level is ideal.


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Separation of human from animal areas is the best way to

minimize disturbances.

Continuous exposure to acoustical levels above 85 dB canhave both auditory and no auditory effects, including

Eosinopenia and increased adrenal weights in rodents.

Reduced fertility in rodents.

Increased blood pressure in nonhuman primates.

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SOUND


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ENVIRONMENTAL CONDITIONS REQUIRED FORENVIRONMENTAL CONDITIONS REQUIRED FOR

LABORATORY ANIMALSLABORATORY ANIMALS

Temperature :Temperature :Humidity :Humidity :

Ventilation :Ventilation :

LightLight--dark circles :dark circles :

LightLight--intensity :intensity :

Light source :Light source :

Noise level :Noise level :

2525CC 22C throughout the yearC throughout the year4545--55% throughout the year55% throughout the year

1616--20 fresh air changes/hour20 fresh air changes/hour

12 hours light and 12 hours dark or 14 hours12 hours light and 12 hours dark or 14 hours

light and 10 hours dark.light and 10 hours dark.

300300--400400 luxlux oneone metremetre above the floor levelabove the floor level

Fluorescent tube lights fixed in such a wayFluorescent tube lights fixed in such a way

to facilitate uniform distribution of light.to facilitate uniform distribution of light.

Should be less than 65 decibels (shouldShould be less than 65 decibels (should

avoid all high frequency sound)avoid all high frequency sound)

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FOOD

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Animals should be fedpalatable,

non-contaminated, and

nutritionally adequate food dailyunless the experimental protocol

requires otherwise.

Diet should be free from heavy

metals (e.g., lead, arsenic, cadmium,

nickel, mercury), naturally

occurring toxins , microbial,chemical and other contaminants.

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Diet Composition for Rodents (Rats,Diet Composition for Rodents (Rats,

Mice & Hamsters)Mice & Hamsters)

Wheat flourWheat flourRoasted Bengal gram flourRoasted Bengal gram flour

Skim milk powderSkim milk powder

CaseinCasein

Refined groundnut oilRefined groundnut oil

Salt mixtureSalt mixture

Vitamin mixtureVitamin mixture

22.5%22.5%60.0%60.0%

5.0%5.0%

4.0%4.0%

4.0%4.0%

4.0%4.0%

0.5%0.5%

Diet Composition for Monkeys, RabbitDiet Composition for Monkeys, Rabbit

& Guinea pigs& Guinea pigs

Wheat flourWheat flour

Roasted Bengal gramRoasted Bengal gram

CaseinCasein

Refined groundnut oilRefined groundnut oil

Salt mixtureSalt mixture

Vitamin mixtureVitamin mixture

Vitamin CVitamin C

61.3%61.3%

28.2%28.2%

1.0%1.0%

5.0%5.0%

4.0%4.0%

0.5%0.5%

50mg/100g diet50mg/100g diet

Scale of DietScale of Diet

RatRat

MouseMouse

HamsterHamster

MonkeyMonkey

RabbitRabbit

Guinea pigGuinea pig

1515--20 g/day20 g/day

5 g/day5 g/day

15 g/day15 g/day

150 g/day150 g/day

5050--100 g/day100 g/day

25 g/day25 g/day

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Equipment preparation a. Cages shoe box, filter top, IVC, MICE,

Hanging, metabolic

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Shoe box habitat

Squirrels habitat


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Filter top cage habitat

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IVC - Individually ventilated cages50


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MI

CE MI

CROENVI

ORNM

ENT ISOLAT

ION COTA

INM

ENT - ERICHM

ENT 51


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Hanging cage

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Metabolic cage 53


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The metabolic cage main features are:

Upper chamber - made ofPC Surface; is smooth, gnaw-proof,

silent.

Two models are available: - for mice and rats up to 300 g, with a

surface of 320 cm2 and a height of 14 cm; - for rats over 300 g, with a

surface of 450 cm2 and a height of 18 cm.

Surface and height are in compliance with the current U.S.A.* rules

and regulations.


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Feeder chamber -

Two-part feeder chamber is located outside cage. Front chamber

catches spilled food so faeces won't be contaminated. Feeder sizes

prevent rodent from nesting or sleeping inside. Available in fivesizes.

Drawer slides out of feeder chamber for easy filling , without

disturbing animal.


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Collection funnel and separating cone - Calibrated to accurately

measure intake. Drain diverts overflow into collection tube so

water can't contaminate urine.

Urine flows along the inside surface of the collection funnel and is

directed by the urine ring directly into the urine collection tube.

A simple twist unlocks either faeces or urine tube. No need todismantle cage or disturb animal.

S t id SS id l t t th h th id l


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Support grid - SS grid lets excreta; pass through the widely

spaced bars; mouse cage includes mouse-sized grid. Handles

permit easy transport of cage without stand.

Water Bottle - Calibrated PC bottle with SS cap and sipper tube.

Water Spillage Tube - Collects any water that animals

spills. Facilitates accurate collection of fluid intake data.


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W t


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Water

Tap, RO, chlorinated, acidified, sterile

Bedding

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Bedding should be absorbent, free

of toxic chemicals or other

substances that could injure animals

or personnel, and of a type notreadily eaten by animals.

Bedding should be removed and

replaced with fresh materials as

often as necessary to keep theanimals clean and dry.

E.g. Wood shaving, chip,

corncob, paper


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Hygiene control 1. Personal

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2. Equipment

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3 Cleaning schedule

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Animal care through experiment

Animal acceptation

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Decontamination Identification


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Animal quarantine

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Animal during experiment

Stress

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Animal after experiment -

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Collection from Animals

Collection of blood from tail vein in rat

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General anesthesia needed small amount: 0.11 ml

75% alcohol cotton ball

for surface

disinfection.

27 G1/2 needle with 1

ml syringe for blood

withdrawal

A vial for blood


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Optimal site of blood withdrawal is

around the distal onethird of the tail

since this part of tail gives better

visualizatio

nof

the veins 68


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Disinfect the tail with 75% alcoholic

cotton ball

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When the needle penetrates the epithelium of

the tail, pull back the plugger a bit to create

negative pressure inside the syringe, then

push the needle in the vein slo

wly until bloo

det into the ea s ace of the nee le hea70


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Pull back the plugger by the ring finger to

withdraw blood from the tail vein

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Using a scalpel to make a small wound on the

tail is also an option for collecting blood

from tail vein72


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Blood can be collected b using a pipette,

after then

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Collection of Blood from Cardiac puncture in Rat

General anesthesia needed in large amount: up to

3% of body weight.

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75% alcohol cotton

ball for surfacedisinfection.

24G needle with

10cc syringe for

blood withdrawal.

15 cc centrifuge

tube for blood

collection


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Disinfect the left thorax with 75% alcoholic

cotton ball

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Search for the point of maximum heart

beat

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Insert the needle straight on the

selected point and withdraw blood by

your left hand77


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Research

Classification Pure research

Applied Research Xenotransplantation

Toxicology TestingCosmetic testing

Drug Testing

Education, Breeding & Defense

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Pure Research (Basic research)

a. Investigates how organisms behave, develop,and function.

b. Uses larger numbers and a greater variety of

animals than applied research Fruit flies,nematode worms, mice and rats together

account for the vast majority, though small

numbers of other species are used, ranging

from sea slugs to armadillos.

c. By this, scientists aim to understand both how

organisms normally develop, and what can go

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E.g. Studies on embryogenesis and developmental biology

where mutants are created byadding transposons into their genomes, or

specific genes are deleted by gene targeting.

Experiments into behavior, to understand how

organisms detect and interact with each other

and their environment (fruit flies, worms, mice,

and rats).

Breeding experiments tostudy evolution and genetics.

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Applied Research a. Applied research aims to solve specific and

practical problems.

b. They are usually carried out in the pharmaceutical

industry, or by universities in commercial

partnerships.

c. May involve the use of animal models of diseasesor conditions, which are often discovered or

generated by pure research programs.

d. Such applied studies may be an early stage in

the drug discovery process.

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E


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E.g..

Genetic modification of animals to study disease.

Transgenic animals have specific genes inserted,

modified or removed, to mimic specific

conditions such as single gene disorder, such

as Huntington's disease

Studies on models of naturally occurring disease

and condition.

Cats are used as a model to develop

immunodeficiency virus vaccines and to

study leukemia because of their natural

predisposition to FELINE LEUKEMIA VIRUS.82

A dill d h l f


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Armadillos and humans are among only a few

animal species that naturally suffer

from leprosy; as the bacteria responsible for

this disease cannot yet be grown in culture,armadillos are the primary source of bacilli usedin leprosy vaccines.

Xenotransplantation

Involves transplanting tissues or organs from

one species to another, as a way to overcome the

shortage of human organs for use in organ

transplants.

Although transplant rejection remains a

problem, recent clinical trials that involved

implanting pig insulinsecreting cells into83

T (


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Toxicology testing (safety testing)

Conducted by pharmaceutical companies testingdrugs, or by contract animal testing facilities.

Tests are conducted without anesthesia,

because interactions between drugs can affecthow animals detoxify chemicals, and may

interfere with the results.

Used t

o

examinef

inished pro

ducts suchas pesticides, medications, food additives,

packing materials, and air freshener.

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The substances are applied to the skin or dripped


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The substances are applied to the skin or dripped

into the eyes.

Injected intravenously, intramuscularly,

or subcutaneously.

Inhaled either by placing a mask over the animals

and restraining them, or by placing them in aninhalation chamber; or administered orally,

through a tube into the stomach, or simply in

the animal's food.

E.g. The LD50 ("Lethal Dose 50%") test is used toevaluate the toxicity of a substance by

determining the dose required to kill 50% of the

test animal population.85

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Cosmetics testing

These are particularly controversial

Tests, which are still conducted in the U.S.,

involve general toxicity, eye and skin irritancy,

photo toxicity (toxicity triggeredby ultraviolet light) and mutagenicity.

Cosmetics testing is banned in the Netherlands,

Belgium, and theU

K.

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D


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Drug testing Metabolic tests how drugs are

absorbed, metabolized and excreted by the bodywhen introduced orally, intravenously,

intraperitoneally, intramuscularly,

or transdermally.

Toxicology tests, which gauge acute, sub-acute,

and chronic toxicity.

Efficacy studies, which test whether

experimental drugs work by inducing the

appropriate illness in animals.

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Education, breeding & defense Animals are also used for education and training;

are bred for use in laboratories; and are used bythe military to develop weapons, vaccines,

battlefield surgical techniques, and defensive

clothing.

Pigs to study the effects of improvised

explosive device explosions on internal organs,

especially the brain.

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Important bodies


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Important bodies

IACUC

USDA

OLAW

AAALAC

Institutional policy

Attending veterinarian / Institutional Animal

care personnel

State statues & local ordinances

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IACUC (Institutional Animal Care and


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Use Committee)

Is an institutional body.

Responsibility for review and oversight of

the institutions program for the humane care

and use of animals.

Supports, facilitates, and promotes ethical

and humane use of animals by upholding the

standards set forth in the federal statutes andregulations, policies and guidelines, and

applicable institutions policies and procedures

concerning animal activities.

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IACUC Charge


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IACUC Charge

Review at least once every six months

institutions animal care facilities, laboratories,use program and areas where animals are used.

Prepare and submit reports ofIACUC program

evaluations and facility inspections to the IO.

Review and investigate concerns involving the

care and use of animals in accordance with

institutions Policy.

Suspend animal activities that are not being

conducted in accordance with applicable federal

regulations and guidelines.

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Animal care duties

Housing

Daily Health Checks

Pathogen Control

Feed, Bedding

Transport

Monitoring of Surgery/Other Procedures92


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Necropsy

Physical Plant Upkeep

Equipment Maintenance

Scientific/Clinical Input on IACUC Review

IACUCs eyes and ears in the Field

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Records


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Records

OLAW (office oflaboratory Animalwelfare )

Annual Report/Renewal of Assurance

Update program changes

Forcause site visits

Will waive notforcause site visits if AAALAC

accredited

U

SDA(U.S. DepartmentofAgriculture)

Annual Report/Numbers of Animals Used

Forcause site visits

Notforcause onsite inspections 2x/annually

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AAALAC(Associationfor assessment & accreditation of

laboratory animalcare )

Annual Report

Update program changes

Onsite visits every 3 years

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AAALAC (Association for assessment


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(

& accreditation of laboratory animal

care )

Is a private, nonprofit organization.

Promotes the humane treatment of animals in

science through a voluntary accreditation

program.

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Functions

Evaluate all aspects of an animal care and use

program.

Evaluate use of an animal in research, education,

testing or breeding.

Basic components of a program that are

evaluated include IACUC policies, animal

husbandry, veterinary care.

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The Principles ofHumane Experimental


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Technique

In 1959, Russell and Burch proposed the conceptof the Three Rs.

REPLACEMENT refers to methods which avoid or

replace the use of animals in an area whereanimals would otherwise have been used.

REDUCTION refers to any strategy that will

result inf

ewer animals being used

REFINEMENT refers to the modification ofhusbandry or experimental procedures to

minimize pain and distress 98

These Rs are widely accepted ethical principles.


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These Rs are widely accepted ethical principles.

Now embedded in the conduct of animalbased

science in Canada and throughout many

countries in the world.

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Conclusion


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Conclusion

With the use, study & proper care of lab animals &

the knowledge gained throu gh research, we can

continue improving the lives & life standard ofnot

only humans, but our pets, wildlife & other animalstoo.

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References


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References

http://en.wikipedia.org/wiki/Animal_testing#Definitions

www. iacuc.org; ALAT Training Manual, page 10

www.fbresearch.org/education/laws.htm

http://www.usda.gov/wps/portal/usda/usdahome?navid=anima

l_production&navtype=RT&parentnav=agriculture

The humane care and treatment of laboratory

animals National Association of Biomedical Research.

Retrieved February 8, 2008.

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www CanadiancouncilofAnimal care com


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www.CanadiancouncilofAnimalcare.com

Ramaswamy et al. ; The Ethics of research involving animals;Nuffield Council on Bioethics; (27)2008: 46 49

W. m. s Russel & R. L. Burch; The principals of humane

experimentaltechnique; ALTEX20 (Suppl1): 6376.

Americans for medical progress www.amprogress.org

Foundation for biomedicalresearch www.fbresearch.org

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Thank You

lab animals - kush

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