P3 Biosafety Lab
A. DESCRIPTION OF THE CBGP P3 LABORATORY FACILITY (A / ES / 13 / I-19)
The facility is designed to prevent that the quarantine pathogens escape from inside. In order to achieve this, the installation has:
a) a) Air recirculation circuit, which is filtered through HEPA filters. When these filters become clogged, the installation must be stopped and desinfected. Then, the filters must be changed and finally the facility may be restart.
b) b) Three zones of negative differential pressure. When any of the doors is opened, the air comes always from outside. Thereby, the probability that the infectious agent escapes is decreased. The three zones are: entrance hall, entrance and exit corridors well differentiated and the laboratory. There is a room with a biological safety hood inside the laboratory. This room must remain closed. All the open manipulations of the infectious agent must be done inside the safety hood.
Notes: the facility is not watertight. Due to the differential pressure, there is always a flow from the outside. This also facilitates the entry of both flying and crawling insects. There is a mosquito trap and cockroach trap in the entrance hall.
B. COMPONENTS OF THE P3 FACILITY (A/ES/13/I-19):
- Exterior entrance door. It is always locked by the key. There is only one copy in use that must be collected and returned from the concierge.
- Entrance and exit doors to hallway and locker room. All doors are interlocked, so none of the doors can be opened if another door is already opened.
- Entrance and exit corridors. These areas act as locker rooms. In addition, they provide another differential pressure zone. The wardrobes have double doors so the objects that have been left athe the entrance, can be collected at the exit.
- Main laboratory. Contains: poyatas, shelves on poyatas, low furniture, fridge, freezer, autoclave and temperature control.
- Phytotron room. It is normally left opened to facilitate the thermal balance. It also contains a -80ºC freezer.
- Biosecurity chamber room. This room contains the chamber and a poyata. It should always be closed, especially when someone is using the chamber.
- Pile and drain It contains a BioWaste that alerts when the tank is full.
- SAS to Exchange objects with the outside.
- Facility controls. They are external, located next to the entrance door. In a normal way, they should not be touched. Unless a maintenance is required, the facility will work continuously and the controls will not be deactivated.
1. Exterior entrance door
2-3. Entry and exit doors, hallway / locker room
4. Main laboratory
5. Phitotron room
6. Biosecurity chamber room
7. Pile and drain
8. SAS (exchang of objects with the outside)
9. Controls of the facility
C. EQUIPMENT OF THE P3 FACILITY (A/ES/13/I-19):
In the facility, there are the following detailed equipment available:
4. Main laboratory (pag. 2)
Electrophoresis tray and power supply Shaker-incubator
Stereomicroscope Campingaz Magnetic stirrer Vortex
Desktop computer Plate reader
Autoclave Fridge-Freezer Liquid effluent decontamination system
Flow hood. Class II UNE EN 12469 Magniflying glass
5. Phitotron room
Freezer Vacuum cleaner
Plants growth chamber
D. OPERATION IN THE P3 FACILITY (A/ES/13/I-19):
- All operations are governed by the principle that the infectious agent must not leave the facility under any circumstances.
- In case of operations with human pathogens, the operator safety also prevails.
- The facility is designed so that the infectious agent cannot be expelled through the air. This is avoid thanks of both, the continuous filtering air through HEPA filters and the pressure barriers that establish one-way flow air INWARD.
- 4. There are only two alternative escape routes for the infectious agent: the objects and the operators.
- Uncontaminated objects can freely enter into the facility.
To remove them, it must be assumed by default, that they are contaminated and they can transmit the infectious agent. Therefore, they can only leave the facility if: :
- They have already been decontaminated.
- They have been isolated in a closed container with a Surface free of infectious agent. Only then, the decontamination of the object can be done outside the facility.
Sterilization. The material will be left in closed autoclave bags for the subsequent sterilization.
- Sterilization in the autoclave of the facility: ATTENTION, this autoclave is not absolutely reliable. It is recommended times of 1 hour. Once it is cooled, it can be passed through the SAS and throw it away.
- Sterilization in an external autoclave. For this, the contaminated material will be left in a double autoclave bag and passed through the SAS. From there, it will go directly to the autoclave with a normal cycle. It should not be sterilized with other materials. It can be thrown away when the cycle ends.
- Chemical decontamination. If the object has been in contact with the infectious agent, it will be treated with a disinfectant and then removed by the SAS.
- SAS. If the object has simply been in the facility and it has not been in contacted with the infectious agent, it will go through the SAS. Any object that does not fit in the SAS, should be placed in a closed plastic container. The walls of this container must be chemically disinfected and the container will removed by the people exit (por example, infected plants must be transported in closed plastic boxes to be carried to the biosafety greenhouse. These boxes must be also chemically disinfected and removed by the exit of the people).
- Sterilization. The material will be left in closed autoclave bags for the subsequent sterilization.
- The facility is designed to contain all kinds of infectious agents. However, the normal use of the facility is focus on working with infectious agents that are not harmful for people. When the infectious agent can be harmful, additional precautions have to be taken. For working with SARS-CoV-2, a specific procedure is established: PROCEDURE WORK IN THE SARS-CoV-2 LABORATORY.
- Entry. Tights and head covers must be used to maintain the cleanliness of the room. The lab coat must be changed in the facility and they can only come out for being washed. Using single-use lab coats above the usual ones it is advisable.
- Exit. We must assume we can carry the infectious agent on the lab coat, gloves, etc. That is why it is necessary all of these clothes leave the facility in closed bags for external treatment (autoclave for disposables and industrial laundry for non-disposables).