1.0 OBJECTIVE: The objective is to design the procedure for autoclave operation.
2.0 PRINCIPLE:
Sterilization refers to the complete killing of all living organisms, including
spores. Common sterilization techniques include the application of wet heat,
dry heat, chemicals, and radiation. Various pieces of equipment are used for
sterilization in laboratory animal facilities, but the autoclave is the primary
means of sterilizing supplies. Despite built-in safeguards, an autoclave
presents the possibility of serious injury to technicians from hot surfaces and
from the release of live steam. It is important, therefore, that laboratory
animal technicians understand the proper operation, limitations, and safeguards
for sterilization by autoclaving.
3.0 RESPONSIBILITY:
It is the responsibility of the Animal house in-charge in conjunction with
the Animal house attendants to ensure that all autoclave equipment are
appropriately monitored, maintained in good working order, and available for
research personnel as requested. The Animal house in-charge is responsible for
ensuring all research and technical staff are adequately trained and
experienced in autoclave sterilization procedures.
4.0 PROCEDURE:
4.1 Autoclave
sterilization time and temperature are dependent on the composition and nature
of the items being autoclaved. Minimum time is usually 15 minutes at 121
0C at 15 lbs pressure.
4.2 Autoclave
was set at 121 0C when sterilizing micro isolator caging. Higher temperatures
can damage polycarbonate caging.
4.3 Items
to be sterilized were loaded into the
autoclave in a manner so that nothing touches the inside of the
chamber.
4.4 Items
to be sterilized were placed into the
autoclave so that steam can uniformly flow between items and so
that no air pockets are formed between or around the items.
4.5 Small
items to be autoclaved should be bagged or wrapped in heat-resistant
plastic, paper, or cloth.
4.6 Sterilized
supplies must be labeled with the date autoclaved/processed. The date of expiration
is determined using the following criteria:
4.6.1
Sterile packs/items should be stored in clean, dry
enclosed cabinets and drawers free from moisture and dust.
4.6.2
Sterile packs prepared by research or technical
staff, when stored in enclosed cabinets/drawers in a dust-free, insect-free,
and well-ventilated area, can be considered sterile for 1 year providing
the packaging remains dry and intact.
4.6.3
Sterile packs prepared by research or technical
staff stored on open shelves have a reduced safe shelf life of 1 month providing
the packaging remains dry and intact.
4.6.4
The manufacturer generally stamps commercially
prepared sterilized packs with an expiration date.
4.6.5
A shelf life of 1 year is suggested for
commercially prepared packs without an expiration date, providing the pack is
stored in an enclosed cabinet/drawer, remains dry, and packaging is intact. Label
with date of receipt.
4.6.6
The quality of the packaging material, storage and
transport conditions, and the amount of handling all contribute to maintain
sterility of the package and its contents.
4.6.7
Any package that is wet, torn, dropped on the
floor, damaged in any way, or past its date of expiration will not be used.
When such events occur, the contents should be removed, repackaged, and
re-sterilized.
4.7 Drinking water is autoclaved by
placing filled water bottles in wire baskets and covering the wire basket with
a suitable cover (e.g., type, or similar material) prior to autoclaving.
Autoclaved water bottles remain covered from the time of autoclaving until the time of
use.
4.8 When
the autoclave cycle is completed and the pressure returns to zero, the door may
be opened.
4.9 Before opening the door, staff members
should wear heat-resistant autoclave gloves. To prevent being scalded from the
steam, move to the opposite side of the door opening. Open the door slowly and once
opened, walk away from the direct line of the steam. After a few minutes, the
autoclave materials can then be removed. Items will still be very hot for
a while. Items should not be used until cool to the touch. Animal feed that has
been autoclaved takes a lot longer to cool down than the container it was
autoclaved in.
4.10 When removing water bottles, let the bottles of water
sit in the autoclave for 10 minutes. The bottles and the water will remain
very hot for a long while. It is recommended that the bottles not be used
until the next day.
4.11 Staff members encountering malfunctions with
the autoclaves should report it immediately to the supervisor, and not attempt
to fix it themselves.
5.0 PRECAUTIONS:
5.1 Technicians
can be seriously injured from hot surfaces and from the release of live
steam. Use of the long-heat-resistant autoclave gloves is necessary.
5.2 If
steam escapes through the gaskets while the autoclave is running, turn the
autoclave off and get the supervisor or next in charge. DO NOT OPEN THE DOOR.
6.0 REFERENCES:
6.1 American
Association for Laboratory Animal Science. Training Manual Series,
Volume II. Laboratory Animal
Technician.
6.2 Maggio-Price L, Shows D, Waggie K, Burich A, Zeng W, Escobar S,
Morrissey P, Viney JL. Helicobacter bilis infection
accelerates and H. hepaticus infection delays the development
of colitis in multiple drug resistance-deficient (mdr1a−/−)
mice. Am J Pathol. 2002;160:739–751. [PMC free article] [PubMed]
6.3 Maher JA, Schub T. Laboratory rodent transportation supplies. Lab
Animal. 2004;33(8):29–32. [PubMed]
6.4 Mangram AJ, Horan ML, Pearson L, Silver C, Jarvis WR. Guidelines for
prevention of surgical site infection, 1999. Infect Control Hosp
Epidemiol. 1999;20:247–278.
6.5 Martini L, Lorenzini RN, Cinotti S, Fini M, Giavaresi G, Giardino R.
Evaluation of pain and stress levels of animals used in experimental
research. J Surg Res. 2000;88:114–119. [PubMed]
END OF DOCUMENTS
You may like to read these links:
1. List of All SOPs and Documents for the Microbiology Laboratory
2. List of All SOPs and Documents for In-vitro Laboratory
3. List of All SOPs and Documents for the Animal House Facility
4. List of All SOPs and Documents for Laboratory Instruments and Equipment
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