EXPERIMENTAL COMMON LABORATORY ANIMAL

INTRODUCTION

Various research domains, including pharmaceutical science, the natural sciences, biology,
medicine, and veterinary medicine, employ experimental or laboratory animals. The majority of
laboratory animals participate in a variety of studies that evaluate the effects of pharmacological
and biologically derived drugs, therapeutically, toxicologically, genetically, and for experimental
surgery using models that replace the human body. A live item (including fish, reptiles,
amphibians, birds, and mammals) utilized in animal experimental study is referred to as a
"laboratory animal." Rodents make up the largest category of experimental animals among all
vertebrates (Pisces, reptile, amphibian, avian, and mammalian species), as they reproduce readily
and have little dietary and environmental requirements.

Rats, mice, rabbits, guinea pigs, and other rodents are considered to be members of the order
"Rodentia," which is part of the class "Mammalia". Although they are not frequently used, other
animals like dogs, cats, pigs, sheep, cattle and apes are still important because of how similar
their bodies are to humans. All of these animals are kept in animal kennels with continuous
access to food and water. Animals must be treated with care to prevent suffering both during and
after the experiment or investigation. Proper design of experiments is necessary to avoid
procedures that ultimately prove unnecessary or useless.

LEGAL REGULATIONS CONCERNING EXPERIMENTAL PROCEDURES WITH

EXPERIMENTAL ANIMALS

The Prevention of Cruelty to Animals Act, 1960, and the Breeding of and Experiments on
Animals (Control and Supervision) Rules adopted under the Act, respectively, govern animal
experimentation in the conduct of medical research and teaching. These are upheld by the
Prevention of Cruelty to Animals Act of 1960's legislative body, the Committee for the Purpose
of Control and Supervision of Experiments on Animals (CPCSEA). According to these
requirements, the involved establishments must register with CPCSEA, complete an IAEC, have
their animal housing facilities inspected, and obtain CPCSEA approval for specific research
projects before starting any animal testing. Furthermore, these Rules also restrict the breeding
and trafficking of animals used in such experiments. The Institutional Animal Ethics Committee
(AEC) of the institutions now has the authority to approve studies on small animals according to
a 2006 revision to the Rules for Breeding of Animals and Experiments on Animals (Control and
Supervision). Only proposals for using big animals in studies must be submitted to CPCSEA for
approval. Therefore, it is crucial that each IAEC member be completely aware of the current
laws and regulations.

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The experimental animals are regulated by the Animal Protection Act dated 21 August 1997
(Law Reports no. 111, item 724), the legal issues are:

1. Research or experimental protocol tests on laboratory animals may be performed only in

scientific institutions approved by CPCSEA under the guidance and supervision of IACE
(Institutional Animals Ethics Committee).

2. Experiments on animals are performed only when they are mandatory for scientific research,
university education, or the protection of the health of people or animals if these objectives
cannot be achieved otherwise as no alternative methods exist (art. 28 of the above Act).

3. The research protocol must be approved by the Local Ethics Committee (IAEC) and
supervised by the National Ethics Committee (CPCSEA) (art. 28). { Institutional Animal Ethics
committee }

4. The husbandry of laboratory animals must be licensed by the Ministry of Agriculture and
Food Economy and the animal house should ensure the conditions appropriate for animal species
(art. 29).

5. Experimental protocol associated with pain must be performed in general or local anesthesia
only once on one individual unless the nature of the experiment requires its repetition on the
same animal. The experiments may be performed without anesthesia only in exceptional cases
when it is obligatory from the scientific point of examination.

ANAESTHESIA

For ethical and scientific reasons, it is imperative that any pain caused by research on animals be
removed or reduced. With the use of general or local anesthetics, the discomfort associated with
experimental surgical techniques may be avoided. Inhaling halothane, injecting pentobarbital, or
combining ketamine and xylazine to cause unconsciousness are common methods of inducing
general anesthesia. However, local anesthetics like lidocaine or procaine only render a portion of
the body insensible. Pre-anesthesia is typically administered to experimental animals before the
administration of anesthesia to reduce side effects and ensure a mild regaining of consciousness.
As an example, anticholinergic medications like atropine, reduce salivation, bronchial mucus
secretion, and other undesirable effects related to the stimulation of the autonomic nervous
system (ANS). Premedication also includes the use of sedatives and tranquilizers, which reduce
tension and aid in regaining awareness. To prevent hypothermia during surgery, the animals
require quiet and temperatures between 35 and 37 degrees Celsius.

EUTHANASIA

Euthanasia is the term used to describe the method typically used in experimental protocols that
allow for the sacrifice of animals for research purposes. The term "euthanasia" refers to the act of
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humanely terminating an experimental animal. There are several procedures, including CO2
exposure, decapitation, cervical vertebral dislocation, overdosing on injectable or inhalation
anesthetics, and cervical vertebral blows. The selection of these euthanasia techniques is made to
guarantee that the animal dies without suffering, loses consciousness, and that the procedure
causes as little stress as possible.

1. ALBINO MOUSE

ALBINO MOUSE
Zoological Name Mus musculus .
Other strains Swiss mice Balb/C, Laca, C-
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Age 1-2 years
Blood volume 70-80 ml/kg
Blood pressure 120/75 mm of Hg
Pulse rate 300-750 per min
Respiration rate 90-150 per min
Body temperature 37.4±0.2°C
Room temperature 24-27°C
Humidity 60-70%
Estrous or oestrous cycle 4-5 days
Mating age 6-8 weeks
Body weight (adult) 23-30 g
Water consumption 5-6 ml/day
Food consumption 5-7 g/day
Urine excretion 1-3 ml/day
Feed Cracked food, shark liver oil, yeast powder, pellets, sesame oil,
fish, some other supplements, etc.
Use Acute toxicity study, an assay of insulin, cancer and genetics
research, screening of chemotherapeutic and teratogenic agents,
the study of analgesics, behavioral study etc. Isolated
preparation: vas deferns, ileum, trachea, fundus, etc.

Note –

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2. ALBINO RATS

ALBINO RATS
Zoological Name Rattus norveglus .
Other strains Sprague Dawley rat,
Wistar rat, Porton,
Biobreeding rat, Long
evans rat, RCS rat,
Zucker rat, Shaking rat
Kawasaki
Age 2-3 years
Blood volume 50-65 ml/kg
Blood pressure 130/90 mm of Hg
Pulse rate 260-450 per min
Respiration rate 80-150 per min
Body temperature 37.5±0.2°C
Room temperature 24-27°C
Humidity 60-70%
Estrous or oestrous 4-5 days
cycle
Mating age 10-12 weeks
Body weight (adult) 180-260 g
Water consumption 35 ml/day
Food consumption 20 g/day
Urine excretion 10-15 ml/day
Feed Cracked food, shark liver oil, yeast powder, pellets, supplements etc
Use Toxicity study, assay of hormones, cancer and genetics research,
screening of chemotherapeutic and teratogenic agents, study of
analgesics, behavioral study, and ulcer, antihypertensive, anti-
diabetics, liver phygiological studies etc. Isolated preparation: vas
defrens, ileum, trachea, fundus, uterus, colon, evaluation of
psychopharmacological agents etc.

Note-

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3. DOMESTIC RABBITS

DOMESTIC RABBITS
Zoological Name Oryctolagus cuniculus .
Other strains Commonly used strains
New Zealand white
strains
Age 4-5 years
Blood volume 45-70 ml/kg
Blood pressure 170/70 mm of Hg
Pulse rate 135 per min
Respiration rate 55 per min
Body temperature 38.3±0.2°C
Room temperature 24-27 °C
Humidity 60-70%
Estrous or oestrous 4-5 days
cycle
Mating age 6-9 months
Body weight (adult) 3-5 kg
Water consumption 200-300 ml/day
Food consumption 100g / day
Urine excretion 200-300 ml/day
Feed Lucerne grass, carrot, bengal gram, wheat bran etc.
Use Pyrogen testing, anti diabetic activity, bioassay of insulin, embryo
toxic study capillary permeability study.

Note -

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4. GUINEA PIG

GUINEA PIG
Zoological Name Cavia Porcellus
Other strains Rex Guinea Pig,
byssinian Guinea Pig,
American Guinea
Pig.
Age 2 years
Blood volume 65-90ml/kg
Blood pressure 170/70 mm/Hg
Pulse rate 250-300 beats/min
Respiration rate 60-110 per/min
Body temperature 37.6-38.9±0.2°C
Room temperature 24-27 °C
Humidity 60-70%
Estrous or oestrous 15-19 days
cycle
Mating age 3-5 months
Body weight (adult) 400-800 gms
Water consumption 250-350 ml/day
Food consumption 30 g/day
Urine excretion 100-150 ml/day
Feed Crushed oats, carrot, Cabbage, Wheat bran etc.
Use Anti asthmatic, local anaesthetics, anti spasmodic agents, anti TB drugs,
Histamine bioassay,Vit C metabolism, Different microorganism
experimental study.

Notes –

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TRANSGENIC AND MUTANT ANIMALS –

These are also known as genetically modified organisms, transgenic animals, mutant animals,
and genetically engineered animals. Transgenic animals are those whose genomes have
purposefully been altered to include a foreign gene, most frequently a mouse gene. These
animals are often produced by microinjecting DNA into the pronuclei of a fertilized egg, which
is then implanted into the ovum of a surrogate mother who isn't pregnant.

Definition:

An animal whose genetic material has been altered by the use of genetic engineering or
recombinant DNA technology.

There are five main categories of transgenic animals.

1. Disease models: Include animals such as Alzheimer's mouse, AIDS mouse, and
OncoMouse™ that are engineered to mimic some aspects of human disease. These animals
provide models for investigating disease mechanisms and potential cures.

2. Transpharmers: Include animals engineered to express protein drugs or antibiotics in their

milk. These animals provide a convenient source for new medications with no animal sacrifice.
This category includes such models as Baby Herman (the world's first transgenic cow, whose
female offspring provide the first successful bovine transpharmer) and the Genzyme goats. These
animals provide new sources for drugs. We normally synthetically produced.

3. Xenotransplanters: Those are engineered to produce organs compatible with humans.

4. Transgenic food sources: That includes animals like Superpig and Superfish that are
engineered to be larger than normal.

5. Transgenics include scientific models: That teach us something new about a specific
protein's function in vivo.

Some of the type of genetically modified animals are mentioned below

1. Transgenic organism:

Synonyms: Mutant

Definition:

Traditionally, an organism with the addition of foreign DNA, whether from the same species or a
different one. More recently the term transgenic has been used to refer to any genetically
modified organism.

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2. Knockout/Knockin:

Synonyms: Global knockout: Constitutive knockout. Definition:

Knockout: The removal or complete disruption of a specific gene in an animal from the
blastocyst stage through adulthood.

Knockin: The introduction of a mutated version of a specific gene in place of the wild-type
version

3. Conditional knockout:

Synonyms: Site-specific knockout, region-specific knockout, cell type-specific knockout.

Definition:

The removal or complete disruption of a specific gene in a manner that controls the cell types
and brain region or site where the disruption occurs. The Cre/loxP system is frequently used to
produce conditional knockouts and in this system, the promoter expressing Cre recombinase will
give rise to the specificity of the excised gene.

4. Inducible knockout:

Synonyms: Time-specific knockout, temporal knockout.

Definition:

The use of a system where the experimenter controls the timing of gene removal. The
tetracycline-inducible system is frequently used to produce animals where a simple injection of
tetracycline will either terminate or initiate gene transcription.

5. RNAi (RNA interference):

Synonyms: siRNA (small interfering RNA).

Definition:

An endogenous system where short sequences of double-stranded RNA molecules induce the
cleavage of matching mRNA resulting in down-regulation of a particular gene. This system can
be manipulated for experimental by using exogenous siRNA to down-regulate a GOI

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6. Chimera:

Synonyms: Mosaic.

Definition:

An animal where individual cells contain genetic material from only one of two potential
lineages. These animals are often produced in the creation of knockout mice where a mutated ES
cell is introduced into the blastocyst containing wild-type ES cells.

7. Forward Genetics/ Reverse Genetics:

Synonyms: Forward genetics Random mutagenesis; Reverse genetics + Targeted mutagenesis.

Definition:

Forward genetics is the examination of the genetic cause of an altered or abnormal phenotype
introduced by a chemical mutagenesis or mutation by irradiation (e.g. phenotype →genotype). In
reverse genetics, a particular gene is altered and the phenotype is investigated (e.g. genotype
phenotype).

8. Phenotype/Genotype:

Synonyms: Not available.

Definition:

Genotype is the specific genetic constitution of an organism including the gene allelic makeup.
Phenotype is the physical trait or characteristic arising from the genotype.

9. Pseudopregnant –

Synonyms: Not available.

Definition – A hormonal state similar to pregnancy that is induced in mice by mating a female
with a vesectomized male. In this state, the uterus is receptive to an implanted embryo.

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