Experimental animals can be divided into four levels according to the experimental animal microbiological control standards:
Primary animal (CV) refers to a general animal whose microorganism is not specifically controlled. It is required to exclude pathogens of zoonosis and a small number of pathogens of experimental animals. In order to prevent infectious diseases, certain measures should be taken when raising and breeding experimental animals, and the results of the tests should be guaranteed to be reproducible (ie regardless of the different operators, at the same time, with the same strain Animals tested in accordance with the prescribed experimental procedures can achieve almost the same results).
Secondary cleansing animals (CL) require the elimination of zoonotic diseases and pathogens of major infectious diseases of animals.
There are no special pathogens (SPF) in the third grade, and it is required to exclude the prescribed pathogens. For sterilization and sterilization, high-efficiency air filter sterilization method, ultraviolet sterilization method, triethylene glycol vapor spray method and lithium chloride can be used.
Aqueous spray method.
Grade 4 sterile animals (GF) or biological animals (GN). Sterile animals are required to have no microorganisms detectable by existing methods. Animals are required to implant one or several known microorganisms on a sterile animal body. On pathological examination,
The four types of experimental animals also have different pathological examination criteria.
The first-level appearance is healthy, and the main organs should not have lesions.
In addition to the primary indicator, the secondary examination revealed no secondary microbial pathogens.
There are no special pathogens in the third grade. No secondary or tertiary pathogens.
Grade 4 contains no secondary or tertiary microbial pathogens. The spleen and lymph nodes are the histological structures of sterile animals.
In summary, there are different requirements for the design and management of animal houses in different levels of experimental animals. Sterile, known bacteria and animals without specific pathogens need to be kept in a sterile or aseptic environment as possible.
Currently known internationally as a barrier environment, a barrier is used to separate animals from the surrounding polluted environment, just as the fetus is in the uterus of the mother. This kind of environment is divided into five categories: isolation system, barrier system, semi-barrier system, open system and laminar flow system from the perspective of controlling microorganisms.
The A isolation system is a system for feeding animals in a container with operating gloves for feeding sterile animals and living animals. The interior maintains a level 100 cleanliness as required by the microorganisms, but the room and operator it is set up do not have to be considered as a sterile room.
The B barrier system uses a sterile clean room of about 10,000 to 100,000 levels as a breeding room, which is mainly used for long-term feeding and breeding of animals without special pathogens. Strict management, such as showering, changing clothes, etc.
The C-half barrier system relaxes the management of people and things in the barrier system when entering and exiting the room. The plane composition is roughly the same as the barrier system.
The D-layer flow rack system cage is placed in clean horizontal laminar air. Often used for small-scale rearing, but there is a risk of contamination during feeding, handling and handling in general rooms. Can be used for the supplement of the semi-barrier.
The E-open system is a system that eliminates pollution in the room for people, things, air, etc., but usually requires some degree of cleaning management.
The requirements for various systems for environmental indicators of laboratory animals are listed below:
Project indicator
Open System (CV) Simple Barrier System (CL) Barrier System (SPF) Isolation System (GF, GN)
Temperature / °C 18~29 18~29 18~29 18~29
Daily temperature difference / °C 8 3 3 3
Relative humidity /% 40~70 40~70 40~70 40~70
Number of air changes / (time / h) 10~15 10~15 10~15
Air flow rate / (m / s) 0.18 0.18 0.18
Pressure difference / Pa 20~50 20~50 20~50
Cleanliness / grade 10,100,100
Number of colonies / [piece / (dish · h)] 12.2 2.45 0.49
Mass concentration of ammonia / (mg / m3) 14 14 14 14
Noise / dB ≤ 60 ≤ 60 ≤ 60 ≤ 60
Illumination / lx 150~300 150~300 150~300 150~300
The purification measures to achieve the animal room environmental indicators are:
A. Air Purification System Because the animal room is a place where experimental animals are active for a long time, indoors cannot be sterilized by fumigation in the breeding conditions. Generally, they can only be washed with a weaker disinfectant. In order to avoid microbial infestation, air purification is the primary condition;
B. The new wind prevents cross-contamination and deodorization are two problems. For this reason, most of the clean air-conditioning systems must adopt a new style of wind, but because the purification system has a large number of air changes and a new wind, the energy consumption problem is outstanding;
C. Deodorization is not only necessary for the external environment and staff, but also for the experimental animals themselves. Because of the microbial contamination in the indoor exclusion zone, the animal's own excreta and the ammonia, hydrogen sulfide, alcohol and other gases generated by the secondary conversion of microorganisms,
It is also an important pollutant.
2. The general environmental factor control range of the animal house
(1) The temperature target value is 21~27 °C. The Pharmacopoeia is specified at 17~28 °C. It can vary depending on the animal species.
(2) Humidity 45% to 55%. The Ministry of Health has a quota of 40% to 60%.
(3) The air flow rate is 10~25cm/s. Avoid direct blowing.
(4) The number of air changes is 6~15 times/h. The fresh air is 2/3.
(5) Positive pressure in the air pressure clean area, negative pressure in the infected area.
(6) Environmental cleanliness level The general animal breeding room is greater than 100000.
(7) Illumination 150~300lx (artificial lighting).
(8) Noise 40~50dB (when there is no animal), 60 dB for animals and less than 70 dB for the Ministry of Health.
(9) Odor ammonia 20ppm.
3. Basic requirements for animal room design
(1) Separation and sanitation
(2) The layout is divided into a preparation area, a feeding area, and an experimental area. Require separate flow of people, logistics, and animals (one-way process);
Purification, sterilization, and pest control are required.
(3) The building requires clean corridors, breeding rooms, contaminated corridors and other clean rooms.
(4) The air conditioning system has controllable temperature and humidity, airflow velocity and distribution, and has a certain amount of ventilation and air pressure.
(5) Illuminated windowless animal rooms use fluorescent lamps, and windowed animal rooms can be fitted with glass windows to filter out UV rays. Request 12
H bright, 12h dark.
(6) The water supply has drinking water and refined water.
Four. Animal room settings
According to the needs of different types and different levels of experimental animals, the animal rooms, activity sites and corresponding auxiliary rooms of the corresponding facilities are established. The site should be chosen to be quiet, clean and free of adverse external influences. Animal rooms must be well-lit, well-ventilated, clean and tidy, without water; ceilings and walls should be easy to clean and disinfect; external walls, roofs, ceilings, doors and windows, and outside pipes must be free of external animals, mosquitoes and other pests. In. The indoor temperature is controlled at 18~29°C, the relative humidity is 40%~70%, and the noise is less than 60dB. The density of animals in the cage should not be too large, and there should be air supply, ventilation, cooling and warming equipment. There must be dedicated sewage and drainage facilities to prevent the spread of pathogens. All kinds of cages should be cleaned and disinfected regularly. The litter should be disinfected by high temperature and high pressure or drug and replaced in time to make the indoor ammonia concentration below 15mg/m3.
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