1. What is the Specific Pathogen Free (SPF) animals?
Reproducibility is one of the most important things in scientific studies. Acquiring low reproducible study result is not only loosing time and budget but also threating executing scientists’ reliability. In the case of the animal experiment, infectious disease is one of the biggest objectives of the reproducibility as a scientific study. If the subject animals got infectious diseases, the following study result should be different from the study, which used infected animals. Therefore, it is very important for experimental animals not to infect any problematic microorganisms to acquire high accuracy and reliable scientific study result.
As a de facto standard, most of the advanced animal experimental research organizations specify such “problematic” microorganisms, caused of not only the infectious diseases and changing the study result but also increasing animal death, reproductive deterioration, and biohazard for researchers, and control their animal room microbiological environment not to have the specified ones. Such microbiological controlled animals are so-called Specific Pathogen Free (SPF) animals.
2. How is CLEA Japan SPF animals?
CLEA Japan, Inc. is the first organization to provide SPF animals in Japan. During our over 50 years’ experience for managing SPF, we have developed our own know-how with various microbiological control technics. Currently, most of the organizations, which have a SPF concept for their animal rooms, can receive CLEA Japan animals directly. (Except they have special study needs or specific microorganisms, which are not existed in Japan.)
The below is the CLEA Japan microbiological monitoring list. We execute the monitoring test for the following microorganisms.
>Click here to see pdf
*CLEA Japan can monitor any microorganisms as your requirement. Please ask us your needs.
3. CLEA Japan methods of SPF environment establishment
■Vinyl Isolator (VI)
VI is a smallest size of SPF keeping unit in CLEA Japan. Currently, we use around 200 small VIs (~35 animals/1VI) and 50 large VIs (~300 animals/1VI). We use them for small-size contract breeding service and caring research subject animals and foundation colonies especially for important animal lines. In addition, we use VI for establishing germ free and gnotobiotic environment, required stricter microbiological control.
Because all caring procedures are completed inside VI, it is easy to minimize the infectious risks.
Requires very high and experienced VI handling and management skills and technics and more time for doing all procedure than other SPF environmental methods. It is also limited the executions such as complicated surgical treatment and etc.
■Individual Ventilated Cage (IVC)
IVC is an SPF condition keeping equipment, able to ventilate clean air by individual lacks or even cages. Animal caring procedures such as changing bedding, diets, and drinking water bottles are executed inside the IVC special caring station/equipment.
The caring technics are easier than VI.
Depend on the number of animals, front-end cost is higher than VI
bioBubble is a assembly type clean room system developed by bioBubble, Inc., U.S. It can be constructed any part of buildings/spaces without any animal facility specialized fanctions for SPF room with matching any space availability. CLEA Japan uses it for storing animals, delivered from outside and better not to share the air for other animal rooms before its health confirmation, and contract research spaces, required to accept researchers and goods of customers frequently.
Any part of buildings can be an SPF animal room.
Higher front-end and running cost. (But it is much lower than constructing whole animal facility)
CLEA Japan calls “BS room” for SPF environment animal room, designed for controlling inside microbiological condition. The rooms are controlled inside air condition with central management system with large HEPA filters that this can cut per animal caring cost by its scale merit. CLEA Japan uses BS rooms for caring production colonies such as the animals for sales.
Beneficial for huge amount of animal caring by scale merit.
High front-end cost (Need to construct as an animal facility from the first.)
4. Tips for CLEA Japan SPF environment management
CLEA Japan manages various Standard Operating Procedures (SOPs) and monitorings in major procedures for maintaining SPF environment. Below are examples.
* These are the SOPs related to keeping SPF condition only.
- SOP of entering/taking-out animals
- SOP of sterilization
- SOP of disinfection
- SOP of entering SPF animal room
- SOP of delivering animals
- SOP of animal production
- SOP of animal management
■Vinyl Isolator (VI)
The difference of VI management from the other systems are 1) all inside VI handlings are done by VI gloves, cannot touch inside things directly and 2) all VI carrying in and out are through a VI specialized sterilization tin. So that these procedures are lot more than other SPF systems and require accuracy and certainty in every steps.
VI management has majorly 2 phases such as an establishment and operation. The summary of the establishment procedures are Leak test 1 > Cage bringing > Filter set > Inside sterilization 1 (Peracetic acid spraying and application) > Inside sterilization 2 (Peracetic acid spraying again with doing leak test 2 together) > Sterilization confirmation test > Start to use. Each leak test and spraying takes 1 day leaving for securing certainty of its sealability and sterilization effect.
The operation phase also requires VI glove handling for all inside manipulations and aseptic manipulation for animals and goods bringing in and out works such as other VI, specialized container, and sterilization tin connection. It is required to prepare accurate and detailed SOPs and strict observance by proficient and experienced personnel.
Expiration date for use:
- Small VI: 10 years
- Large VI: 5 years
- HEPA filter of Small VI: 3.5 years
- Side attached filter of Sterilization tin: 9 months
bioBubble maintains an SPF environment by air flow that it is not sealed from outside, so there are different biological control methods from other SPF systems.
Ensure filter confirmations:
bioBubble needs to maintain air filters strictly. It is essential if filters are surely creating clean air sufficiently. For example, there are 2 major maintenance procedures such as pre-filter cleaning and filter wind speed confirmation.
Caution of personnel traffic lines and procedure sequences:
Because it is not sealed from outside, it is required to comply the “Working from clean to dirty” policy, common rule for animal facility process priority. It means personnel should not go backward from outside to the clean side and disinfect all required things before entering other place in each time. For example, personnel requires to change used gloves, disinfect hands and sole of the shoes by ethanol spray, and blow dust by air spray gun to move into other section each time. In addition, personnel should strictly comply executing SOPs of bioBubble entrance and even disinfect before getting out from clean side in case of failing to keep SPF condition unknowingly.
Caution of entering goods and equipment:
There are bioBubble specialized transfer containers for bringing necessary goods inside. The goods are put into the container and autoclaved with it. Then the container is connected to bioBubble connecting sleeve. The inside of the connecting sleeves are sprayed ethanol from a spraying hole of the sleeve. The goods are brought into inside bioBubble after taking enough time to disinfect inside sleeve completely. With the sterilization effect confirmation by autoclave and ethanol, it is required to be very careful if whole bringing procedure is correctly proceeding.
Inside air of BS room is segregated from outside so it is not so difficult to keep SPF condition if procedures of goods bringing and personnel entering are correctly executed. The major cautions of keeping SPF are as below:
1. Facility deterioration check
2. Sterilization quality check
3. Avoid bringing by personnel
CLEA Japan covers the item 1 and 2 by routing confirmation procedures with making checklists and reports of the related as below:
- Daily report of temperature and humidity
- Sterilization management report (for Diets)
- Sterilization management report (Autoclave execution)
- Animal transfer report
- Environmental confirmation checklist
- Facility/equipment maintenance report
- Facility checklist for holiday
About the item 3, CLEA Japan makes detailed SOP of entering animal room and observance the processes following the SOP strictly. In addition, personnel skin should be covered all over the body when they are inside BS animal rooms even the skin part is carefully disinfected.
■ Question / Inquiry
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5. Related products / Equipment / Facility
Autoclave is an essential equipment for keeping SPF environment. Autoclave can sterilize necessary goods for animal care inside SPF room such as diets, drinking water, beddings, cleaned cages, and etc. by high pressure and temperature steam. There are various types and outside boiler may be necessary depend on the products.
All goods must be sterilized once these should be brought inside SPF animal rooms and diets, beddings, and drinking water are included. Especially diets must keep these nutrients enough to keep animals healthy even these are autoclaved. CLEA Japan supplies diets, which can be surely keeping enough nutrients for the needs of the animal health status for studies after autoclaving.
* CLEA Japan can supply autoclaved diets by autoclaving at our Thailand facility. Please contact us about our autoclaving service.
Air Shower is a device installed at the doorway to a clean room for removing dusts and germs attached to entering personnel. CLEA Japan installed it in front of immunodeficiency animal rooms to avoid bringing specific infectious microorganisms, which are additionally specified for immunodeficiency animals.
■Vaporized Hydrogen Peroxide (VHP) sterilizer
VHP sterilizer makes VHP gas inside its chamber and sterilizes necessary goods for SPF animal rooms. It is used for the goods which cannot be autoclaved such as electronic equipment, plastic products, and etc.
■Question / Inquiry
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5. Please contact us if you have any question about SPF environment.
- About SPF room establishment (Recommended installation equipment by animal colony size)
- Advises of SPF animal room management
- Sales of the products for keeping SPF environment
- Sales of SPF animals