PRESENTED BY: S.S.CHAUDHARI

(M.PHARM 1ST SEMISTER)

DEPARTMENT OF PHARMACOLOGY

R C PATEL COLLEGE OF

PHARMACY

SHIRPUR

CPCSEA GUIDELINES FOR LABORATORY ANIMAL CARE AND

HANDLING

What Are The Animal Laws?2

The Constitution of India

The Indian Penal Code, 1860

The Criminal Procedure Code, 1973

The Prevention of Cruelty to Animals Act, 1960

The Wildlife Protection Act, 1972

The Police Acts

The Municipal Corporation Acts.

GOAL (1)

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To promote the humane care of animals used inbiomedical and behavioral research and testing.

To provide specifications that will enhance animal well being.

To provide quality in gaining advanced biological knowledge that is relevant to humans and animals

GUIDELINES (1,2,3,4)4

VETERINARY CARE

Adequate veterinary care must be provided and is the responsibility of a veterinarian.

Daily observation.

ANIMAL PROCUREMENT

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All animals must be acquired lawfully as per the CPCSEA guidelines.

A health surveillance program for incoming animals should be carried out to assess animal quality.

Inspect for compliance with procurement specifications.

QUARANTINE6

An effective quarantine minimizes the chance for introduction of pathogens into an established colony.

A minimum duration of quarantine - Small lab animals - 1 week and

- Larger animals - 6 weeks

STABILIZATION AND SEPARATION7

Newly received animals should be given a period for physiologic, psychologic and nutritional stabilization before their use.

Duration for stabilization will depend on the type of animal, transportation and intended use.

Physical separation of animals by species is recommended.

SURVEILLANCE, DIAGNOSIS, TREATMENT AND CONTROL OF DISEASE

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Observe for signs of illness, injury, or abnormal behavior.

Unexpected deaths and signs of illness should be reported.

If animals are known to be exposed to an infectious agent the group should be kept intact and isolated during the process of diagnosis, treatment, and control.

Diagnostic clinical laboratory may be made available.

ANIMAL CARE AND TECHNICAL PERSONNEL9

Employ people trained in laboratory animal science.

They should be provide for both formal and on-the-job

training.

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Annexure – 7 CERTIFICATE COURSE FOR LABORATORY

ATTENDANT (Basic Education: 8th standard)

Introduction Animals rooms Handling of animals Personal hygiene Emergency situations

PERSONAL HYGIENE11

It is essential to maintain a high standard of personal cleanliness.

Decontaminate clothing exposed to potentially hazardous microbial agents or toxic substances.

Use disposable gear.

No permission to eat, drink, smoke or apply cosmetics in animal rooms.

ANIMAL EXPERIMENTATION INVOLVING HAZARDOUS AGENTS

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Institutional Biosafety Committee.

The procedures must be reviewed by both the Institutional Biosafety committee and Institutional Animal Ethics Committee (IAEC).(Annexure – 8)

Annexure – 813

Institutional Biosafety Committee (IBSC)

Institutional Biosafety Committee (IBSC) is to be constituted in all centers engaged in genetic engineering research and production activities.

The Committee will constitute the following.

(i) Head of the institution or his nominee(ii) 3 or more scientists engaged in DNA work or molecular

biology with an outside expert in the relevant discipline.

(iii) A member with medical qualification- Biosafety officer (in case of work with pathogenic agents/large scale used.)

(iv) One member nominated by DBT

MULTIPLE SURGICAL PROCEDURES ON SINGLE ANIMAL

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Multiple surgical procedures not to be practiced unless specified in a protocol only approved by the IAEC.

DURATIONS OF EXPERIMENTS

No animal should be used for experimentation for more than 3 years unless adequate justification is provided.

PHYSICAL RESTRAINT15

Brief physical restraint can be accomplished manually or with devices.

Prolonged restraint of any animal should be avoided unless essential to research objectives.

Less restrictive systems, such as the tether system or the

pole and collar system should be used when compatible with research objectives.

The following are important guidelines for the use of restraint equipments:

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Not be used simply as a convenience in handling or managing animals.

Minimum time. Should be given training to adapt to the equipment.

Observe the animal at appropriate intervals.

Veterinary care should be provided if lesions or illness associated with restraint are observed.

PHYSICAL PLANT17

The physical condition and design of animal facility should be well planned and properly maintained.

PHYSICAL RELATIONSHIP OF ANIMAL FACILITIES TO LABORATORY

Isolated far away from human habitation.

Place animal housing areas adjacent to or near laboratories but separated

FUNCTIONAL AREAS18

Sufficient animal area required to:

Ensure separation of species or isolation of individual projects when necessary;

Receive, quarantine, and isolate animals;

Provide for animal housing.

PHYSICAL FACILITIES19

(1)Building material (7) Corridor

(2)Animal room doors (8) Exterior windows

(3)Floors

(4)Drains

(5)Storage areas

(6)Experimental area

ENVIRONMENT20

(a) TEMPERATURE AND HUMIDITY CONTROL

18 to 29°C temperature range 30% to70% relative humidity

(b) VENTILATION

(c) POWER AND LIGHTING21

A time-controlled lighting system

Emergency power should be available in the event of power failure

(d) NOISE CONTROL

ANIMAL HUSBANDRY22

(a) CAGING OR HOUSING SYSTEM:

Provide adequate space

Comfortable environment

Escape proof enclosure that confines animal safety

Easy access to food and water;

Provide adequate ventilation

Meet the biological needs of the animals

Keep the animals dry and clean

Annexure – 3AMinimum floor area recommended for laboratory animals (based on their weight/size and behavioral activity)23

Animal Weight in Floor area Cage height gram (cm2) (cm)

MICE <10 38.7 upto15 1.6 upto25 7.4 >25 96.7 12

RATS <100 109.6 upto200 148.3 upto300 187.0 upto400 258 upto500 387 >500 >=451.5 14 GUINEA PIG <350 387 >350 >=651.4 18

HAMSTERS <60 64.5 upto80 83.8 upto100 103.2 >100 122.5 12

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(b) SHELTERED OR OUTDOOR HOUSING:Accessible to all animals, sufficient ventilation, and prevent

build up of waste materials and excessive moisture.

(c) SOCIAL ENVIRONMENT:Whether the animals are naturally territorial or communal.

FOODAnimals should be fed palatable, non-contaminated, and

nutritionally adequate food daily.

BEDDING25

DESIRABLE CRITERIA

AbsorbentFree of toxic chemicalsAmmonia bindingSterilizableEasily stored UncontaminatedNontoxicDisposable by incinerationReadily available

WATER26

Continuous access of fresh, potable, uncontaminated drinking water.

Periodic monitoring of microbial contamination.

SANITATION and CLEANLINESS

Cleaning with detergents and disinfectants.

Wire – bottom rodent cages should be washed at least every 2 weeks.

Disinfection of cages by rinsing at a temperature of 82.20C (180 F).

WASTE DISPOSAL AND PEST CONTROL27

Remove regularly and frequently. Animal tissues, carcasses, and hazardous wastes should

be lined with leak – proof, disposable liners.

EMERGENCY, WEEKEND AND HOLIDAY CARE

Animals should be cared by qualified personnel every day, including weekends and holidays.

RECORD KEEPING28

The Animal House should maintain following records:

Animal House plans Animal House staff record All SOPs for animals Breeding, stock, purchase and sales records Minutes of institutional Animals Ethics Committee

meetings Records of experiments Death Record Clinical record Water analysis report

STANDARD OPERATING PROCEDURES (SOPs)29

The Institute shall maintain SOPs describing procedures / methods of

Animal Husbandry MaintenanceBreeding Animal house microbial analysis Experimentation records.

30TRANSPORT OF LABORATORY ANIMALS

The main considerations for transport of animals are:

Mode of transport ContainersAnimal density in cages Food and water during transit Protection from transit infections Injuries and stress

ANNEXURE – 4REQUIREMENTS FOR TRANSPORT OF LABORATORY ANIMALS BY ROAD, RAIL AND AIR

SpeciesSpecies Mouse Rat Hamster G.Pig Rabbit Cat Dog Monkey

Max. no of animal/ cage

25 25 25 12 2 1 or 2 1 or 2 1

Material used in transp-ort

Metal Cardboard ,Synthetic material

Metal Cardboard, Synthetic material

Metal Cardboard, Synthetic material

Metal Cardboard, Synthetic material

Metal Cardboard, Synthetic material

Metal Metal Bamboo/wood/ metal

Space/ animal (cm2)

20-25 80-100 80-100 80-100 1000-1200 1400- 1500

3000 2000-4000

Min. height of box (cm)

12 14 12 15 30 40 50 48

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ANAESTHESIA(4)32

Sedatives, analgesics and anaesthetics should be used to control pain or distress under experiment.

NARCOSIS

RELAXATION ANALGESIA

ANNEXURE – 5COMMONLY USED ANAESTHETIC DRUGS FOR LABORATORY ANIMALS

Drugs(mg/kg)

Mouse Rat Hamster Rabbit Cat Dog Monkey Guinea pig

KETAMI-NE

22-24 i/m

22-24 i/m

-- 22-24 i/m 30 i/m 30 i/m 15-40 i/m 22-24 i/m

PENTO-BARBITONE

35 i/v

50 i/p

25 i/v

50 i/p

35 i/v

-

30 i/v

40 i/p

25 i/v

-

20-30 i/v

-

35 i/v

-

30 i/v

40 i/p

THIOPENTONE SODIUM

25 i/v

50 i/p

20 i/v

40 i/p

20 i/v

40 i/p

20 i/v 25 i/v

-

25 i/v

-

25 i/v

60 i/p

20 i/v

55 i/p

URETHANE - 0.75 i/p - 0.75 % 1.25 i/v

1.50 i/p

1.0 i/v 1.0 i/v 1.5 i/p

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EUTHANASIA34

(a) Death, without causing anxiety, pain or distress with minimum time lag phase.

(b) Minimum physiological and psychological disturbances.

(c) Compatibility with the purpose of study and minimum emotional effect on the operator.

(d) Location should be separate from animal rooms and free from environmental contaminants.

ANNEXURE – 6 EUTHANASIA OF LABORATORY ANIMALS

(A – Methods Acceptable for species of animals indicated NR – Not Recommended) Species Mouse Rat Hamster Guinea

pigRabbit Cat Dog Monkey

a) PHYSICAL METHODSElectrocutionExsanguinationDecapitationCervical dislocation

NR NR A A

NRAAA

NRAAA

NRANRNR

NRANRNR

NRANRNR

NRNRNRNR

NRNRNRNR

b) INHALATION OF GASESCarbon mono oxideCarbon di oxideCO2+ChloroformHalothane

AAAA

AAAA

AAAA

AAAA

AAAA

AAAA

ANRNRA

ANRNRA

c) DRUG ADMINISTRATIONBarbiturate overdoseChloral hydrate overdoseKetamine overdoseSodium Pentothol

(IP)NRIM/IPIP

(IP)NRIM/IPIP

(IP)NR(IM/IP)IP

(IP)NRIM/IPIP

IV,IPIVIM/IVIV

IV,IPIVIMIV

IV,IPIVIMIV

IV,IPIVIM/IVIV

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LABORATORY ANIMAL ETHICS36

All scientists working with laboratory animals must have a deep ethical consideration for the animals they are dealing with.

TRANSGENIC ANIMALS

Transgenic animals are those animals, into whose germ line foreign gene(s) have been engineered.

Knockout animals are those whose specific gene(s) have been disrupted leading to loss of function.

MAINTENANCE37

Special care has to be taken with transgenic/gene knockout animals

Prevent cross breeding with other animals.

Knockout animals should be maintained in clean room environment or in animal isolators.

DISPOSALThe transgenic and knockout animals should be first

euthanized and then disposed off.

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BREEDING AND GENETICS

For initiating a colony, the breeding stock must be procured from CPCSEA registered breeders or suppliers.

39Experimentation on animals in course of medical

research and education is covered by provisions of the Prevention of Cruelty to Animals Act, 1960 and the Rules under the Act of 1998 and 2001. (10)

The Prevention of Cruelty to Animals Act, 1960 provided for constitution of CPCSEA (11)

The Experiments on Animals (control and supervision) Amendments Rules (1998) gave power to CPCSEA to make rules. (11)

OPERATIONAL GUIDELINES IN CPCSEA(10)40

Main activities

(a) Registration of establishments for Breeding of animals: (b) Registration of establishments for experiments on

animals:

(c) Approval of Animal House Facilities:

(d) Permission of Committee for Conducting Experiments:

(e) Recommendation for import:

2. Regulatory Mechanism :41

2.1 Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA) To ensure that animals are not subjected to unnecessary pain or suffering before, during or after performance of experiments on them.

2.2 Sub-Committees of the CPCSEA:1) Sub-Committee on Large Animals (SCLA).

2) Sub-Committee for selection of CPCSEA Nominees on Institutional Animals Ethics Committees.

2.3 Institutional Animal Ethics Committee (IAEC)42

Every establishment is required to constitute an Institutional Animal Ethics Committee.

Every IAEC shall include;A biological scientist, Two scientists from different biological disciplines,A veterinarian involved in the care of animals, A scientist in charge of animals facility of the

establishment concerned,A scientist from outside the institute, A non-scientific socially aware member andA representative or nominee of the CPCSEA. A specialist may be co-opted while reviewing special

projects.

KEY ROLE43

To assess proposal to use animal for scientific purposes, including teaching and decide whether1)The use is justified2)Welfare of animal is protected

FOR ANIMAL USE TO BE JUSTIFIED,THE AEC MUST

Be convinced that the benefits from the use outweigh the potential costs to the animal

Be assure that ‘3 Rs’ of the animal use must be justified

2.4 Member Secretary, CPCSEA:44

Member Secretary, is responsible for registration of establishment engaged in Breeding of Animals and also in Experimentation on Animals.

Application for permission for conducting experiments are brought before the CPCSEA and SCLA by MS.

He has also been authorized to constitute the Inspection Teams for inspection of Animal House Facilities.

3. Standing time tables of meetings of the regulator/expert body:

COMMITTEES CPCSEA

SCLA

IAEC

Sub-Committee for selection of CPCSEA Nominees on IAEC

Member Secretary, CPCSEA

DAYSQuarterly as on 20thDec, 20th March, 20th June, 20th Sep

10thof every alternate month

Vary as per need

Once in 6 month

No requirement of time table45

4. Nomination of Co-chair/Vice chair of the regulator/expert body for presiding over the meeting in the absence of the chair

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5. Ground Rules :

(1)Registration of establishment for Breeding of animals & for experiments of animals:

The Member-Secretary would take a decision on any application for Registration within two months of its receipt.

Incase the application can not be decided within two months, it will be brought to the notice of CPCSEA at its ensuing meeting.

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(i) The application should be accompanied by a bank draft ofRs.1,000/- in favor of CPCSEA as registration fee.

(ii) The establishment should have constituted an IAEC before applying for registration.

(iii) Before taking the decision regarding Registration, a spotinspection of the organization would be necessary.

(ii) Approval of Animal House Facilities:

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(a) Detailed inspection by a Inspection team. As per the existing practice the following is the composition of inspection team:

(i) CPCSEA’s Nominee

(ii) Expert Consultant, CPCSEA

(iii) Person in charge of another animal house in the sameState, where the institution carrying out experiment islocated.

(iii) Permission of Committee for Conducting Experiments

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(a) No proposal for Animal Experimentation would be entertained, unless the Animal House Facility of the concerned establishment has been approved by competent authority.

(b) The approval for experimentation on small animals will be accorded by IAEC.

(c) Approval for experimentation on Large Animals will be

considered by SCLA on the recommendation of IAEC.

PRINCIPLES FOR UTILIZATION AND CARE OF ANIMALS TO BE USED IN TESTING, RESEARCH AND TRAINING

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Principle No.1“Experiments on animals” (including experiments involving

operations on animals) may be carried out for:The purposes of advancement by new discovery Knowledge which is expected to be useful for saving or for

prolonging human life or alleviating suffering For combating any disease, whether of human beings,

animals or plants

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Principle No.2.Animals lowest on the phylogenetic scale which may give

scientifically valid results should be used for any experimental procedure.

Experiments should be designed with the minimum number of animals to give statistically valid results at 95% level of confidence.

Alternatives not involving animal testing should be given due and full consideration and sound justification provided if alternatives, when available, are not used.

“Cockroach“ (Invertebrates) < Birds < Rodents < Canines < Bovine/Equines <Primates (e.g. Rhesus macaque)< More evolved Primates (e.g. chimpanzee)

Principle No.353

Proper use of animals in experiments and avoidance or minimization of pain and suffering inflicted on experimental animals should be an issue of priority for research personnel.

All scientific procedures that may cause more than momentary or slight pain and/or suffering should be performed with appropriate sedation, analgesia or anesthesia.

Principle No.454

Persons engaged in animal experimentation have a moral responsibility for the welfare of the animals after their use in experiments.

Principle No.5The living conditions of animals should be appropriate for

their species and contribute to their health and comfort.

REFERENCES 55

(1) Committee for the purpose of control and supervision on experiments on animals, CPCSEA guidelines for laboratory animal facilities, Chennai: 3-36.

2) Goyel R K., et al.(2006) Different laboratory animals and their applications, Practical in pharmacology, B.S.Shah prakashan,6:7-29.

3) Kulkarni S K, Common laboratory animals (1999) Handbook of experimental pharmacology, Vallabh prakashan, Delhi 3:11-18.

4) Wolfensohn Sarah, Lloyd Maggie. (2000) Introduction to the principles of animal care and use. Handbook of laboratory animal management and welfare 1:55-69

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5) Foster H L., Small J D., Fox J G.(1982) The mouse in biomedical research, experimental biology & oncology, Academic press ,London 2:1-9

6) Waynforth H B., Flecknell P A.(1999) Administration of substance, anaesthesia & post operative care Experimental and surgical technique in rat, Academic press, London,2:1-147

7) McLeod L J., Pharmacological experiments on intact preparations. Edixrurgh & London:1-32

8)Vogel H G.,(2000) Anesthetics, Drug discovery and evaluation In pharmacological

assays, Springer. 2:587-658.

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9)Report of consultative group review of the norms and practices for regulation of animal experimentation (2004), Ministry of environment and forest, Govt. of India:1-18

10) Operational guidelines for observance of good practices in CPCSEA:1-11

11)Subramoniam A., Gitanjali B.,(2003) CPCSEA- A double edged sword, Indian Journal of Pharmacology 35: 71-72

QUESTIONS 58

1)Discuss about CPCSEA regulations for laboratory animal care.

2)CPCSEA regulations for animal care.3)Discuss the regulations for laboratory animal care and

ethical requirements.

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