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TITLE: STANDARD OPERATING PROCEDURE
SUBJECT: Entry and Exit Procedure in Central Animal House Facility
SOP number: 001 Version number: 01 Pages: 03
Issue date: 02/01/2017 Effective date: 09/01/2017 Next review date: 08/01/2020
Name Signature Date
Prepared By Dr. Shonima P.
Reviewed By Dr. A. Hannah Rachel Vasanthi
Approved By Dr. C. Thirunavukkarasu
TITLE: STANDARD OPERATING PROCEDURE
SUBJECT: Entry and Exit Procedure in Central Animal House Facility
SOP number: 001 Version number: 01 Pages: 03
Issue date: 02/01/2017 Effective date: 09/01/2017 Next review date: 08/01/2020
I. OBJECTIVE
The objective of this SOP is to minimize the risk of the spread of pathogens.
II. SCOPE
This SOP describes the procedure to enter and exit the Central Animal House Facility,
Pondicherry university.
III. RESPONSIBILITY
Principal investigator and their research staff, animal care staff, veterinary care staff,
facility supervisor and anyone entering in the animal facility.
IV. ABBREVIATIONS
a. CAHF: Central Animal House Facility
b. PI : Principle Investigator
c. PPE : Personal Protective Equipment
d. SOP : Standard Operating Procedure
V. PRECAUTIONS
To minimize the generation of aerosols and contaminating dust particles, the PPE shall be
removed in the following sequence, starting from the dirtiest PPE: shoe covers, apron, head
cap, gloves, face mask.
Personnel should not be permitted to eat, drink, smoke or apply cosmetics and perfumes inside
animal facility.
Personnel should keep their mobile either switched off or in silent mode before entering inside
the animal facility.
Charging of mobiles inside the facility is not allowed.
In the event of an emergency inside CAHF use the extension phone to call upon the emergency
personnel. A list of emergency numbers to be contacted is displayed outside the CAHF
entrance.
VI. PROCEDURE
4.1. CAHF Entry Procedure
Step in the central disinfectant footbath at the CAHF entrance.
The personnel shall sign into the access log book provided at the entrance of CAHF, stating
the date, name, department, purpose of visit, in time and out time.
The personnel can place their personal belongings into the place provided in the area.
The personnel shall follow the visual instructions on the wall on the PPE.
The PPE required are: lab coat, latex/nitrile gloves, face mask, head cap, anti-slip
shoe cover.
The belongings of the researchers or PI should be kept in the rack provided. Investigators
should not bring their bags/laptops inside the facility.
If the personnel have items to bring into the facility, these items shall be transferred into the
boxes. Each item shall be decontaminated with 70% ethanol or Isopropyl alcohol before
placing them into the box.
The box shall then be decontaminated in the same manner.
4.2. CAHF Exit Procedure
If the personnel wish to return to continue their work for the day, he/she shall exit through the
entrance.
If the personnel have already entered the dirty area, he/she shall not return to the clean area
for the day.
The personnel shall remove the PPE and discard each piece directly into the trash bin located
back side of the facility and exit through the back door.
The investigators should finish the work with animals as early as possible and exit from the
facility.
A separate sitting area is made available for the animal caretakers.
VII. REFERENCES
Guide for the care and use of laboratory animals, 8th Edition, The National Academic press.
VIII. REVISION HISTORY
Nil
TITLE: STANDARD OPERATING PROCEDURE
SUBJECT: Personal Protective Equipment
SOP number: 002 Version number: 01 Pages: 03
Issue date: 02/01/2017 Effective date: 09/01/2017 Next review date: 08/01/2020
Name Signature Date
Prepared By Dr. Shonima P.
Reviewed By Dr. A. Hannah Rachel Vasanthi
Approved By Dr. C. Thirunavukkarasu
TITLE: STANDARD OPERATING PROCEDURE
SUBJECT: Personal Protective Equipment
SOP number: 002 Version number: 01 Pages: 03
Issue date: 02/01/2017 Effective date: 09/01/2017 Next review date: 08/01/2020
I. OBJECTIVE
The objective of this SOP is to prevent the spread of pathogens from or to the animal
facility through fomites.
II. SCOPE
This SOP describes the proper attire required for working in designated animal areas.
III. RESPONSIBILITY
Principal investigator and their research staff, animal care staff, veterinary care staff,
facility supervisor and anyone entering in the animal facility.
IV. ABBREVIATIONS
PI : Principal Investigator
SPF: Specific Pathogen Free
V. PRECAUTIONS
Do not wear lab coat (which is used in animal house) outside the facility.
Cover legs, feet, and arms when working with chemical, infectious, or radioactive
hazards.
Ensure that head cap covers all hair.
VI. PROCEDURE
1.6.1. Outer protective clothing:
It is essential that the animal care staff maintain a high standard of personal cleanliness.
Central Animal House facility is provided with appropriate personnel protective equipment.
Wear dedicated outer protective clothing: lab coat, latex/nitrile gloves, face mask, head cap,
anti-slip shoe cover, in animal facility, especially when in contact with infected animals, their
waste or body fluids, radioactive or chemical hazards, SPF animals maintained in a barrier
facility.
Wear a separate lab coat inside the quarantine area.
Restrict clothes to a given group of animals with the same health status (e.g., same room,
same herd, similar infectious status).
Change gloves frequently to minimize fomite contamination within the animal room.
Wash hands thoroughly with soap and water, or use an alcohol sanitizer at the end of the
procedure or upon leaving the facility.
Wear face mask to avoid risk of splashing or droplet exposure to mucous membranes.
Wear protective glasses or a face shield whenever there is a risk of exposure or
physical injury to the face/eyes.
Use biosafety cabinet while dealing with infectious agent or harmful chemicals or fumes.
Wear disposable shoe covers and head cap whenever working with infected animals or their
waste or radioactive hazards.
VII. REFERENCES
Guide for the care and use of laboratory animals, 8th Edition, The National Academic press.
Standard Operating Procedure for Institutional Animal Ethics Committee, CPCSEA.
VIII. REVISION HISTORY
Nil
TITLE: STANDARD OPERATING PROCEDURE
SUBJECT: Animal Quarantine and Acclimatization
SOP number: 003 Version number: 01 Pages: 03
Issue date: 02/01/2017 Effective date: 09/01/2017 Next review date: 08/01/2020
Name Signature Date
Prepared By Dr. Shonima P.
Reviewed By Dr. A. Hannah Rachel Vasanthi
Approved By Dr. C. Thirunavukkarasu
TITLE: STANDARD OPERATING PROCEDURE
SUBJECT: Animal Quarantine and Acclimatization
SOP number: 003 Version number: 01 Pages: 03
Issue date: 02/01/2017 Effective date: 09/01/2017 Next review date: 08/01/2020
I. OBJECTIVE
The objective of this SOP is to ensure that all animals that enter the facility do not carry
pathogens that may spread diseases to the other animals.
II. SCOPE
This SOP describes the procedures for quarantine and acclimatization of rodents in CAHF,
Pondicherry University.
III. RESPONSIBILITY
Principal investigator and their research staff, animal care staff, veterinary care staff,
facility supervisor.
IV. ABBREVIATIONS
CAHF: Central animal House Facility
PI : Principal Investigator
SOP : Standard Operating Procedure
V. PROCEDURE
One month prior to the start of the experiment, PI/researcher should inform the Veterinarian
regarding the purchase of the animals.
Based upon the availability of quarantine room, holding room, cages and water bottles, room
will be allotted for the conduct of the study.
Before receiving the consignment vendor shall submit the health certificate stating the
physical fitness of the animals. Serology report for the consignment also should be submitted
along with the health certificate.
On receipt of animals from outside facility, animals should be kept in quarantine room.
The animals are isolated from the rest of the facility for at least 7 days until they appear to be
free of infectious diseases or pathogens.
Animals acquired by the CAHF have to acclimatize for a period of 3-5 days to the
environment before being used after quarantine.
If the rodents received are sick/unwell/injured, they should be isolated from the other animals
as soon as possible.
Rodents due for acclimatization will be housed normally in their respective holding rooms.
However, they will not be used for any experiment or procedure until the acclimatization
period of 3-5 days.
The attending veterinarian should inspect the entire consignment for any abnormality, injury,
mortality and take necessary steps regarding.
The technicians shall receive the animals through the back door of the CAHF and shift into
the quarantine room.
Transfer the animals into clean cage.
The bedding material, feed and water bottle shall be provided with the cage inside the
quarantine room.
Details of the animals shall be written on the Quarantine card so that the PI/researcher can
identify the animals later on.
Routine health monitoring shall be done by the attending veterinarian during the quarantine
period.
PIs/Researchers shall locate the cages of the rodents from the quarantine cage cards and will
be allowed to use the animals after the quarantine/acclimatization period.
Before releasing animals from quarantine for study the Veterinarian should assess the health
status of the entire stock of animals and certify that animals are healthy and can be used for
experimental purposes.
VI. REFERENCES
Guide for the care and use of laboratory animals, 8th Edition, The National Academic press.
Standard Operating Procedure for Institutional Animal Ethics Committee, CPCSEA.
VII. REVISION HISTORY
Nil
TITLE: STANDARD OPERATING PROCEDURE
SUBJECT: RODENT HUSBANDRY
SOP number: 004 Version number: 01 Pages: 05
Issue date: 02/01/2017 Effective date: 09/01/2017 Next review date: 08/01/2020
I. OBJECTIVE
The objective of this SOP is to confirm the welfare of the animals maintained in the facility.
II. SCOPE
This SOP describes the basic procedures for routine care and husbandry of rodents.
III. RESPONSIBILITY
Animal care staff, facility supervisors and Veterinarian.
IV. ABBREVIATIONS
CAHF: Central Animal House Facility
IAEC: Institutional Animal Ethics Committee
PI : Principal Investigator
SOP: Standard Operating Procedures
V. PRECAUTIONS
Feed delivery methods should ensure that all animals are able to access feed for a sufficient
period of time while minimizing feeding aggression and nutrient loss.
Albino rodents are more susceptible to phototoxic retinopathy hence lighting should be
appropriate for those animals also.
VI. PROCEDURE
4.1. Observation of animals
Observe all rodents daily for general condition, injury and illness and to ensure they have
sufficient food and water, including weekends and holidays.
4.2. Emergency and holiday care
Provide routine care and husbandry for all rodents on weekends and holidays.
Make provisions for emergency care by providing both work hours and after hours contact
information for PI’s and their staff. Provide special requests or instructions if needed.
In the event of an emergency inside CAHF use the extension phone to call upon the emergency
personnel. A list of emergency numbers to be contacted is displayed outside the CAHF
entrance.
4.3. Animal identification methods:
Identify all cages with a cage card.
During quarantine period quarantine cage card shall be used and once study initiate study
cards shall be placed and which include details such as name of the investigator, IAEC
protocol number, species, strain, sex and number of animals.
Identify individual animals as needed in accordance with rodent identification methods such
as ear punching, ear tags, tattooing.
4.4. Feeding and Watering
Feed and water are provided ad libitum unless otherwise stated by the researcher.
Animals should be fed palatable, uncontaminated diets that meet their nutritional and
behavioral needs.
Feed should be stored in a type-appropriate manner to preserve nutritional content, minimize
contamination, and prevent entry of pests.
Feeding methods and frequency vary widely depending on the species and age/size of species
Commercial diets from recognized suppliers is been used in CAHF and the storage time
should be based on manufacturer recommendation.
Store open feed bags in leak-proof containers with tightly fitting lids.
Animals should have access to potable, uncontaminated drinking water according to their
particular requirements.
Periodic monitoring for pH, hardness, and microbial or chemical contamination may be
necessary to ensure that water quality is acceptable.
4.5. Social and Environmental Enrichment
Group-house rodents whenever possible
When animal is housing single provide environmental enrichment.
4.6. Environment and Environmental control
Maintain room temperatures between 18 and 24° C.
Maintain relative humidity between 30% and 70%.
Maintain a daily log of room temperature and humidity.
4.5. Bedding
Bedding is used to absorb moisture, minimize the growth of microorganisms, and dilute and
limit animal’s contact with excreta.
Use enough bedding to keep the rodents clean and dry until the next scheduled change.
Animals should have adequate bedding substrate and/or structures
for resting and sleeping. For many animals (e.g., rodents) contact bedding expands the
opportunities for species-typical behavior such as foraging, digging, burrowing, and
nest and it absorbs urine and feces to facilitate cleaning and sanitation.
If provided in sufficient quantity to allow nest building or burrowing, bedding also
facilitates thermoregulation.
4.6. Cage changing and Water bottle Changing
Change cages twice weekly to keep the animals clean and dry and provide a healthy
environment. Wash water bottles twice weekly to keep the drinking water safe and healthy.
4.7. Lighting
Lighting should be diffused throughout an animal holding area and provide sufficient
illumination for the animal’s welfare while permitting good housekeeping practices,
adequate animal inspection including for the bottom-most cages in racks, and safe
working conditions for personnel. Light in animal holding rooms should provide for both
adequate vision and neuroendocrine regulation of diurnal and circadian cycles.
Light levels of about 325 lux (30-ft candles) approximately 1 meter above the floor appear to
be sufficient for animal care.
All animal holding rooms should follow a 12 hour light-dark cycle.
4.8. Pest control
Animal house should be constructed in such a way to prevent, control, or eliminate the
presence of or infestation by pests.
4.9. Housing and Space requirements
All animals should be housed under conditions that provide sufficient space as well as
supplementary structures and resources required to meet physical, physiologic, and behavioral
needs.
An appropriate housing space or enclosure should also account for the animals’ social needs.
Social animals should be housed in stable pairs or groups of compatible individuals unless
they must be housed alone for experimental reasons or because of social incompatibility.
The primary enclosure should provide a secure environment that does not permit animal
escape and should be made of durable, nontoxic materials that resist corrosion, withstand the
rigors of cleaning and regular handling, and are not detrimental to the health and research use
of the animals.
Provide cages that are appropriate in size for the number of rodents housed.
VII. RFERENCES
Guide for the care and use of laboratory animals, 8th Edition, The National Academic press.
Standard Operating Procedure for Institutional Animal Ethics Committee, CPCSEA.
VIII. REVISION HISTORY
Nil
TITLE: STANDARD OPERATING PROCEDURE
SUBJECT: Routine Health Monitoring of Animals in Central Animal House Facility
SOP number: 005 Version number: 01 Pages: 03
Issue date: 02/01/2017 Effective date: 09/01/2017 Next review date: 08/01/2020
Name Signature Date
Prepared By Dr. Shonima P.
Reviewed By Dr. A. Hannah Rachel Vasanthi
Approved By Dr. C. Thirunavukkarasu
TITLE: STANDARD OPERATING PROCEDURE
SUBJECT: Routine Health Monitoring of Animals in Central Animal House Facility
SOP number: 005 Version number: 01 Pages: 03
Issue date: 02/01/2017 Effective date: 09/01/2017 Next review date: 08/01/2020
I. OBJECTIVE
The objective of this SOP is to ensure the welfare of the animals by the animal caretakers.
II. SCOPE
This SOP describes the procedures for the daily care of the animals housed at CAHF. It also
describes the procedure for monitoring the animal holding.
III. RESPONSIBILITY
Animal caretakers, technician and Veterinary care staff.
IV. ABBREVIATIONS
CAHF : Central Animal House Facility
SOP : Standard Operating Procedure
V. PRECAUTIONS
If an entire room of animals is known to be exposed to an infectious agent, the group should
be kept intact and isolated during the process of diagnosis, treatment and control.
Do not house adult male animals in groups’ in order to avoid fighting.
VI. PROCEDURE
The animals housed in the CAHF must check once daily.
Observe all animals daily for clinical signs of illness, injury or abnormal behavior, including
weekends and public holidays.
Observe each cage and check for adequate feed and water, cage cleanliness, dead animals,
overcrowding.
Record the room temperature and room relative humidity.
Observe the animals in quarantine daily twice for mortality, sickness, fighting or injury.
Isolate the dominant animal when found fighting inside the group housed cage.
When animal found sick isolate the animal and check the health status.
During post-operative recovery or when animals are ill or have a physical deficit, more
frequent observations are done and recorded.
Animals that show signs of a contagious disease should be isolated from healthy animals.
VII. REFERENCES
Guide for the care and use of laboratory animals, 8th Edition, The National Academic press.
VIII. REVISION HISTORY
Nil
TITLE: STANDARD OPERATING PROCEDURE
SUBJECT: Guidelines for Blood Collection Volumes and Frequency
SOP number: 006 Version number: 01 Pages: 06
Issue date: 02/01/2017 Effective date: 09/01/2017 Next review date: 08/01/2020
Name Signature Date
Prepared By Dr. Shonima P.
Reviewed By Dr. A. Hannah Rachel Vasanthi
Approved By Dr. C. Thirunavukkarasu
TITLE: STANDARD OPERATING PROCEDURE
SUBJECT: Guidelines for blood collection Volumes and frequency
SOP number: 006 Version number: 01 Pages: 06
Issue date: 02/01/2017 Effective date: 09/01/2017 Next review date: 08/01/2020
I. OBJECTIVE
The objective of this SOP is to ensure well-being of laboratory animals during the collection
of blood from different routes.
II. SCOPE
This SOP describes recommended volumes and frequency for blood collection for commonly
used laboratory animal species.
III. RESPONSIBILITY
Principal Investigators and their staffs, veterinary care staffs.
IV. ABBREVIATIONS
IAEC : Institutional Animal Ethics Committee
SOP : Standard Operating Procedure
V. PRECAUTIONS
Considerations regarding the species to be bled, the size and health status of the animal, the
quantity and type of sample needed (i.e. whole blood, serum, etc.) and the frequency of
sampling, should all be taken into account.
Remember to calculate beforehand the minimum amount of blood necessary to perform all
tests and assays. It is recommended to take no more blood than is absolutely necessary.
Do not puncture a site presenting inflammation or a hematoma.
Limit the number of punctures to four punctures per day with no more than two punctures per
site.
If too much blood is withdrawn too rapidly or too frequently without replacement
(approximately 2% of the animal’s body weight at one time), the animal may go into
hypovolemic shock.
VI. PROCEDURE
The acceptable quantity and frequency of blood sampling in all species is dependent upon
the total blood volume of the animal.
Observe animals prior to sample collection for weakness, illness, dehydration, obesity, or
anemia.
Monitor the animal during and after blood sampling for signs of shock.
4.1. General Principles of Blood Collection in Animals
The method of blood collection should be described in the protocol approved by the IAEC.
It should be least painful and stressful
Blood sample collection from animals may be under anesthesia or without anesthesia
depends upon the route of blood collection.
All nonterminal blood collection without replacement of fluids is limited up to 10% of total
circulating blood volume in healthy, normal, adult animals on a single occasion and collection
may be repeated after 3-4 weeks. Sometimes in particular cases repeated blood samples are
required at short intervals, a maximum of 0.6ml/kg/day or 1.0% of an animal’s total blood
volume can be collected in every 24 hours.
Blood samples are collected using the following techniques:
Blood collection without anesthesia :
Saphenous vein (Rat and Mice)
Dorsal pedal vein (Rat, Mice)
Blood Collection under anesthesia :
Tail vein (Rat and Mice)
Orbital sinus (Rat and Mice)
Jugular vein (Rat and Mice)
Marginal ear vein/artery (Rabbit)
Procedure for saphenous vein blood sample collection
Animal should be restrained manually or by using a suitable animal restrainer.
Remove the hair at the site of blood collection using hair removal cream or electric trimmer.
Hind limb should be immobilized and slight pressure is applied above the knee joint.
20G needle is used to puncture the vein. After puncturing, a syringe with needle is used to
collect blood.
To avoid pain local anesthetic cream can be used on the collection site 30 minutes before the
procedure.
Continuous sampling shall be avoided and not more than three attempts are made.
Collecting more than four samples in a day (24 hours) is not advisable.
Procedure for blood collection from dorsal pedal vein
Dorsal pedal vessels are located on top of the foot so hind foot around ankle is held for that.
The foot is cleaned with antiseptic solution and dorsal pedal vein is punctured with 23/27G
needle.
Then the drops of blood that would appear on skin surface are collected.
Procedure for blood sample collection from tail vein
For collection of blood a 23G needle inserted into tail vein, if the vein is not visible, tail
is dipped into warm water(40ᵒC). The tail should not be rubbed from the base to tip.
To avoid pain local anesthetic cream can be used on the collection site 30 minutes before the
Procedure.
In some cases if blood collection is difficult, cut open 0.5 to 1 cm of surface of skin and the
vein is pricked with blood lance or needle.
Procedure for blood sample collection orbital sinus
This method is also called periorbital, posterior orbital and orbital venous plexus bleeding.
The animsl skin around the eye is pulled taut and scuffed with thumb and fre finger of the
nondominant hand.
The capillary tube is inserted into 30 degree angle to nose at the medial canthus of the eye.
To puncture the tissue slight thumb pressure is enough and enter the plexus or sinus.
Blood will come through the capillary tube. Remove the capillary tube after collection of
required volume of blood.
Bleeding can be stopped by giving a gentle finger pressure using a sterile cotton.
Procedure for blood collection from jugular vein
The jugular veins appear blue in colour and is found 2 to 4 mm lateral to sternoclavicular
junction. The neck region of animal is shaved and kept in hyper extended position.
A 25G needle is inserted in the caudocephalic direction and blood is withdrawn slowly to
avoid collapse of these blood vessels.
Animal has to be handled carefully and not more than 3 to 4 mm of needle is to be inserted
into the blood vessel.
If the attempt to collect fails, one more attempt can be made if there is no bleeding otherwise
the needle is slowly removed and the site is monitored for bleeding.
To stop bleeding finger pressure is applied.
Number of attempt is limited to three.
Apply local anesthetics cream 30 minutes prior to sampling.
Procedure for blood collection from marginal ear vein or artery
For blood sample collection from rabbits this method is adopted.
The animal should be placed in a restrainer.
Ear is cleaned with antiseptic solution and local anesthetic cream is applied on collection site
10 minutes prior to sampling. If requires vasodilators also can be applied to dilate the blood
vessels.
26G needle may be used to collect blood from marginal vein.
Sterile cotton is kept on the collection site, after collecting blood and finger pressure is applied
to stop the bleeding.
4.1. Maximum volumes and recovery periods:
Percent of blood volume collected
in a single sampling
Recovery period
(weeks)
7.5% 1
10% 2
15% 3
4.2. Blood Volume by species:
Species Circulating Blood
Volume (ml/kg bw)
7.5%
(ml/kg bw)
10%
(ml/kg bw)
15%
(ml/kg bw)
Mice 72 5.4 7.2 14.4
Rat 64 4.8 6.4 12.8
Rabbit 56 4.2 5.6 11.2
VII. REFERENCES
Guide for the care and use of laboratory animals, 8th Edition, The National Academic press.
VIII. REVISION HISTORY
Nil
TITLE: STANDARD OPERATING PROCEDURE
SUBJECT: Maintenance of Cleanliness at Central Animal House Facility
SOP number: 007 Version number: 01 Pages: 03
Issue date: 02/01/2017 Effective date: 09/01/2017 Next review date: 08/01/2020
Name Signature Date
Prepared By Dr. Shonima P.
Reviewed By Dr. A. Hannah Rachel Vasanthi
Approved By Dr. C. Thirunavukkarasu
TITLE: STANDARD OPERATING PROCEDURE
SUBJECT: Maintenance of Cleanliness at Central Animal House Facility
SOP number: 007 Version number: 01 Pages: 03
Issue date: 02/01/2017 Effective date: 09/01/2017 Next review date: 08/01/2020
I. OBJECTIVE
The objective of this SOP is to ensure that all animal caretakers do the same procedures to
effectively maintain the CAHF, Pondicherry University.
II. SCOPE
This SOP describes the steps taken to maintain cleanliness and the health status of the animals.
III. RESPONSIBILITY
Animal caretakers.
IV. ABBREVIATIONS
CAHF : Central Animal House Facility
SOP : Standard Operating Procedure
V. PRECAUTIONS
The animal caretakers should sanitise the gloves with 70% ethanol or hand sanitizer
between cages. For cages suspected of a contamination, a change of gloves should be done before
and after handling the cage.
VI. PROCEDURE
The animal caretakers and researchers shall keep the rooms neat and tidy as they are shared
between users.
All floors, including animal rooms, procedure rooms, and corridors shall be swept and mopped
with suitable solution daily.
All door handles shall be sanitized weekly.
All general waste bins shall be checked daily. When full, the trash bag shall be removed and
replaced with a new trash bag.
The foot bath and wheel bath area must be cleaned and filled every alternate day,do ens
ure that soil, manure, bedding material should not come into the foot bath.
VII. REFERENCES
Guide for the care and use of laboratory animals, 8th Edition, The National Academic press.
VIII. REVISION HISTORY
Nil
TITLE: STANDARD OPERATING PROCEDURE
SUBJECT: Euthanasia of Laboratory Rodents
SOP number: 008 Version number: 01 Pages: 03
Issue date: 02/01/2017 Effective date: 09/01/2017 Next review date: 08/01/2020
Name Signature Date
Prepared By Dr. Shonima P.
Reviewed By Dr. A. Hannah Rachel Vasanthi
Approved By Dr. C. Thirunavukkarasu
TITLE: STANDARD OPERATING PROCEDURE
SUBJECT: Euthanasia of Laboratory Rodents
SOP number: 008 Version number: 01 Page number: 03
Issue date: 02/01/2017 Effective date: 09/01/2017 Next review date: 08/01/2020
I. OBJECTIVE
The objective of this SOP is to ensure humane euthanasia of laboratory rodents.
II. SCOPE
The scope of this SOP is to describe the different acceptable methods for humane euthanasia
of laboratory rodents.
III. RESPONSIBILITY
It is intended for all personnel trained in euthanasia of rodents.
IV. ABBREVIATIONS
CAHF: Central animal House Facility
SOP: Standard Operating Procedure
V. PRECAUTIONS
Euthanasia techniques should minimize distress and anxiety experienced by the animal
before loss of consciousness.
The method of euthanasia should not cause any adverse emotional effect on the operator.
VI. PROCEDURE
Euthanasia means “good death” and refers to the intentionally ending a life with the purpose
of relieving pain and suffering.
Euthanasia techniques should result in rapid loss of consciousness and quickly followed by
cardiac or respiratory arrest with the final loss of brain function.
The procedure should be carried out quickly and by a painless method in an atmosphere free
from fear or anxiety.
Euthanasia in CAHF is done in necropsy room which is located away from animal rooms and
free from physiological and psychological disturbances.
The method of euthanasia is selected based on the nature of study, species of animal to be
euthanized.
The euthanasia techniques described in this SOP are :
Non-physical Methods: Overdose of inhalant anesthetic and Overdose of injectable
barbiturate.
4.1. Overdose of inhalant anesthetic
The animal(s) shall be placed into a euthanasia chamber/cage.
The animal(s) shall be exposed to the vaporized isoflurane until respiration stops for at least
2 minutes.
4.2. Overdose of injectable barbiturate
Sodium pentobarbitone (300mg/ml) can be used for humane euthanasia.
120mg/kg of the solution is recommended for intraperitoneal (IP) injection.
The personnel shall scruff/secure the animal by hand or using an animal restrainer securely
when injecting the euthanasia solution into the animal.
Respiration should cease for at least 2 minutes to ensure the death of the animal.
VII. REFERENCES
Guide for the care and use of laboratory animals, 8th Edition, The National Academic press.
Standard Operating Procedure for Institutional Animal Ethics Committee, CPCSEA.
VIII. REVISION HISTORY
Nil
TITLE: STANDARD OPERATING PROCEDURE
SUBJECT: Operation of Biosafety Cabinet
SOP number: 009 Version number: 01 Pages: 03
Issue date: 02/01/2017 Effective date: 09/01/2017 Next review date: 08/01/2020
Name Signature Date
Prepared By Dr. Shonima P.
Reviewed By Dr. A. Hannah Rachel Vasanthi
Approved By Dr. C. Thirunavukkarasu
TITLE: STANDARD OPERATING PROCEDURE
SUBJECT: Operation of Biosafety Cabinet
SOP number: 010 Version number: 01 Pages: 03
Issue date: 02/01/2017 Effective date: 09/01/2017 Next review date: 08/01/2020
I. OBJECTIVE
The objective of this SOP is to ensure the primary barrier protection for individuals working
with biohazardous materials.
II. SCOPE
The scope of this SOP is to describe the appropriate procedures for working in a biosafety
cabinet.
III. RESPONSIBILITY
These procedures must be followed by all who use biosafety cabinet in CAHF.
IV. ABBREVIATIONS
BSC : Biosafety Cabinet
CAHF: Central Animal House Facility
SOP : Standard Operating Procedure
V. PRECAUTIONS
Formulation of drugs and dosing procedures that could create airborne biohazards should
always be performed in a BSC as it protects laboratory workers and the environment from
aerosols or droplets that could spread biohazardous material.
Wear proper PPE before starting the procedure.
Avoid rapid movements of arms/hands inside BSC.
VI. PROCEDURE
Operate cabinet blowers for five minutes before beginning work to allow the cabinet to purge
or remove particulates from the cabinet.
Disinfect the work area. Wipe the work surface, interior walls and surface of the window with
a suitable disinfectant such as 70% ethanol.
Introduce only those items that are required to perform the procedures and arrange in a logical
order. Each item should be wiped with antiseptic solution prior to placing it in into the cabinet
in order to reduce the introduction of contaminants. The flow of work should proceed across
the work surface from clean to contaminated areas.
Similarly pipette tips, discard trays containing disinfectant, biohazard bags, sharps containers,
etc. should be placed to one side of the BSC.
Perform procedures slowly by moving items in and out of the cabinet using a straight in and
out motion and avoiding side to side motions. After placing arms/hands inside the BSC,
manipulations should be delayed to permit the cabinet to stabilize and allow the flow of air to
remove surface contaminants from your arms/hands.
When work is completed all items within the cabinet should be wiped down with disinfectant
and removed from the cabinet. Do not use the interior of the BSC as a storage area since stray
organisms may become trapped and contaminate future experiments. The interior surfaces of
the cabinet should also be cleaned with a disinfectant. Let the blowers operate for five minutes
with no activity inside the cabinet to purge the cabinet of contaminants.
If UV lights are used, they must be tested yearly to ensure that the appropriate wavelength for
decontamination is emitted (254nm). Please note that dust particles on the light and on the
surface of the BSC will inhibit decontamination. If not used properly, UV light can also cause
serious eye and skin injury. Its better to use antiseptic solution for decontamination instead of
UV light.
Investigators should remove their gloves and thoroughly wash their hands exiting the
laboratory.
VII. REFERENCES
National Sanitation Foundation (www.nsf.org). NSF-49 Safety Standard.
Guide for the care and use of laboratory animals, 8th Edition, The National Academic press.
Standard Operating Procedure for Institutional Animal Ethics Committee, CPCSEA.
VIII. REVISION HISTORY
Nil
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