Q．What is BioSafety Level (BSL)?
A．BSL is an indicator of the strictness of safety management in laboratories and facilities that handle microorganisms and pathogens. The BSL of a laboratory that handles a pathogen is determined according to the risk posed by that pathogen. BSL is divided into four levels, with the higher the number, the stricter the safety measures. BSL-1 and BSL-2 are the basic measures for containing pathogens, and BSL-4 is the highest level of safety control. BSL-4 pathogens include Ebola virus and Lassa virus.
Q．Are BSL-4 facilities operational in Japan?
A．The Murayama Branch of the National Institute of Infectious Diseases (NIID) (in Musashimurayama City, Tokyo) has a BSL-4 facility mainly for the diagnosis of infectious diseases. , It was legally designated as a BSL-4 facility (in legal terms, a “Facility for Possessing Specified Class I Pathogens, etc.”) in August 2015 by the Ministry of Health, Labour and Welfare based on the Infectious Diseases Control Law. It was also designated in July 2019 for the import of specified Class I pathogens, etc. It is currently in operation. It is the only BSL-4 facility that is already in operation.
Q． How many BSL-4 facilities are in operation around the world?
A．There are more than 54 BSL-4 facilities in operation or scheduled to be in operation in over 20 countries around the world*. There are BSL-4 facilities in India, Taiwan, China and South Korea in the Asian region. The United States, Germany, Italy also have BSL-4 facilities, and some of them are located on university campuses. New BSL-4 facilities are currently being planned or constructed around the world. Most of the new BSL-4 facilities that have been built in recent years are suit-type laboratories.
*Virus, Vol. 72, No. 2, pp139-148, 2022
Q．What types of BSL-4 laboratories are there other than suit-type (Positive Pressure Suits(PPS))?
A．There are two types of BSL-4 laboratories: glove box type and PPS type. A glove box is a sealed box in which samples are handled by the hand inserted through a rubber glove. The box is also called a Class III cabinet. BSL-4 pathogens are contained within the box in a glove box type laboratory. In a suit-type laboratory, the experimenter wears a high-airtight, PPS and performs the experiment in a worktable (Class II cabinet) with an open front. The laboratory is required to be equipped with a high-airtight structure and a chemical shower for when leaving. The pathogens are contained not only in the Class II cabinet but also in the laboratory itself.
Q．Is the BSL-4 facility at Nagasaki University currently in operation?
A．At present, the BSL-4 facility does not possess specified Class I pathogens, etc. At present, we are in the process of conducting research and development using pathogens of BSL-3 or lower as preparation for research and development using specified Class I pathogens, etc.
Q．What are the plans for using the BSL-4 facility at Nagasaki University?
A．We are conducting research to develop diagnostic techniques and therapeutic strategies for infectious diseases caused by BSL-4 pathogens, as well as research to clarify the mechanisms by which diseases occur. Another important objective is to train researchers who will work to control infectious diseases. We also plan to conduct tests on samples from patients suspected of being infected with BSL-4 pathogens in the future.
Q．Please tell me the structure, building area, and total floor area of the BSL-4 facility at Nagasaki University.
A．The structure is reinforced concrete, with some parts made of steel framed reinforced concrete. The building area is about 1,300 square meters, and the total floor area is about 5,100 square meters.
Q．When do you plan to bring in Specified Class I Pathogens, etc. into the BSL-4 facility?
A．It is supposed to take a considerable amount of time to be able to actually transport Specified Class I Pathogens, etc. into the facility. Before the transport, we need coordination with the institution which already holds and manages Specified Class I Pathogens, etc. and arrangements for transportation. It is also necessary to obtain designation or approval from the Minister of Health, Labour and Welfare for transportation. The timing of the delivery and the specific kinds of pathogens etc., cannot be forementioned.
Q．How are Specified Class I Pathogens, etc. transported?
A．When transporting Specified Class I Pathogens, etc., the transport must be entrusted to a transport company with the necessary expertise and technology. The pathogens must be carried out under the supervision of the Public Safety Commission, in accordance with the Infectious Diseases Control Law.
Q．Why is it necessary to conduct research on Specified Class I pathogens, etc. , such as the Ebola virus?
A．When we consider the expansion of areas where infectious diseases are prevalent due to climate change as well as the increase in international mobility due to the growth of international exchange, we believe that it is extremely important to respond to infectious diseases that are prevalent around the world to ensure the safety and reassurance of our country. The 2014-16 Ebola outbreak in West Africa spread to Europe and the United States. Marburg Virus Disease was first discovered in Germany. Crimean-Congo hemorrhagic fever has also occurred in Europe. There has been one case of a person returning to Japan who developed Lassa fever. Therefore, these should not simply be dismissed as endemic diseases of remote areas of Africa. It would not be surprising if these infectious diseases were to enter Japan at any time in the future.
Research on specified class I pathogens may be applicable to other infectious disease control measures. For example, the development of therapeutic drugs and vaccines for Ebola hemorrhagic fever was immediately put to use in the outbreak of COVID-19. This has been a significant help in the fight against the COVID-19 pandemic.
Specifically, Remdesivir, a drug developed for Ebola, has been shown to be a very effective antiviral drug against COVID-19 (as long as it is administered early). Establishment of new modalities has also contributed to the rapid development and practical application of COVID-19 vaccines.
As mentioned above, research into the Ebola virus and other Class I pathogens is also expected to be of great use in the fight against other infectious diseases. Thus, we believe it is important to conduct research into Specified Class I Pathogens, etc., in a BSL-4 facility.
Q．If an unknown infectious disease were to emerge, would a BSL-4 facility be necessary to deal with it?
A．In the event of an outbreak of an unknown infectious disease, the properties of the pathogen are unknown in the early stages. Thus, the risk of handling the pathogen in the laboratory is assessed based on the characteristics of the pathogen, the situation (transmissibility, severity, etc.) of the infectious disease in the community and the characteristics of similar pathogens. As a result, if it is determined that the highest level of safety management is necessary to ensure the containment in the laboratory and the safety of workers, it will be necessary to handle the unknown pathogen in a BSL-4 facility. Therefore, BSL-4 facilities are also necessary for dealing with unknown infectious diseases (sometimes referred to as “Disease-X”) that may arise in the future.
Q．Does Nagasaki University have a history of research on infectious diseases and the potential to do so?
A．Nagasaki has functioned as an international city open to the world since the Edo period. On the other hand, they were also the first to suffer the effects of the emerging infectious diseases of the time, such as cholera, measles, smallpox and influenza, which were brought to Japan from overseas via Dejima, the artificial island in Nagasaki used as a trading post. The Nagasaki Igaku Denshusho (later changed to the Kojima Yojosho and now the Nagasaki University School of Medicine), which was the modern Western-style hospital and medical school, was founded in 1857. The treatment and prevention of infectious diseases such as cholera formed an important part of its educational and medical activities. This infectious disease research has now been taken over by the Nagasaki University School of Medicine and the Nagasaki University Institute of Tropical Medicine. In this way, we believe that Nagasaki University has the potential and many years of accumulated experience in research and education in diagnosis, treatment and prevention methods, making it a base for infectious disease research not only in Japan but also around the world.
Q．Are there any examples of how Nagasaki University's infectious disease research potential has been put to use in the fight against COVID-19?
A．Nagasaki University has promptly developed a rapid testing system for COVID-19 using the fluorescence LAMP method, a gene amplification technique. This testing system can detect the CCPID-19 genes from patients in a short time. The device used in this system is lightweight, compact, and easy to use, so it can be used not only in a building of medical institutions but also can be carried to areas where infection is spreading. Since this testing system was developed early in the pandemic, it was possible to quickly test all passengers on the cruise ship “Costa Atlantica” when it arrived at Nagasaki Port. Consequently, the outbreak on the ship was brought under control without any deaths.
Q．What kind of research do you plan to conduct once the BSL-4 facility is fully operational?
A．We are aiming at making progress in research and development into highly lethal infectious diseases and emerging infectious diseases that are not being researched on a global scale, and also creating world-leading results in research into Specified Class I Pathogens, etc. For this goal, we plan to conduct research into the analysis of the mechanisms of infection and onset of diseases such as Ebola hemorrhagic fever and Crimean-Congo hemorrhagic fever, the development of therapeutic drugs and vaccines, and responses to unknown infectious diseases that may emerge in the future. Until Specified Class I Pathogens, etc. are introduced and the BSL-4 facility begins fully operational, we will conduct preparatory experiments using BSL-3 or BSL-2 pathogens for the above research.
Through the above research, we also plan to train personnel who can handle Specified Class I Pathogens, etc. Specifically, we aim to establish a systematic education and training program for the first time in Japan for researchers and those involved in the maintenance and management of facilities who will use the Positive Pressure Suit (PPS)-type BSL-4 facility. Then we will train personnel with extensive knowledge and experience in the safe handling of Specified Class I Pathogens, etc.
Q．It says that experiments using BSL-3 or BSL-2 pathogens will be conducted before possession of specified Class I pathogens, etc., but what is the significance of this?
A．The BSL-3 or lower pathogens used in the experiments here are similar in nature to BSL-4 viruses. For example, they have similar characteristics in terms of how they invade cells, how they multiply, and how the human immune system is involved in this process. In addition, the process of analyzing BSL-3 or lower pathogens also serves to familiarize researchers with the use of equipment and procedures. Starting with experiments using BSL-3 or BSL-2 pathogens is extremely important as a preparation before handling BSL-4 pathogens.
Q．Is there any risk of the viruses in the air in the BSL-4 laboratory leaking out?
A．BSL-4 facilities have a structure in which a sealed BSL-4 laboratory is installed inside a building. The laboratory is kept at a lower pressure than the outside. Thus the air inside the laboratory does not flow out to the outside. Exhaust air from the laboratory is discharged outside after being filtered through double high-efficiency (HEPA; High Efficiency Particulate Air) filters to remove even the finest particles.
Q．I heard that BSL-4 laboratory are highly airtight, and negative pressure is kept in the laboratory. What does negative pressure mean? Also, please tell us about the mechanism that prevents the leakage of pathogens by negative pressure.
A．Negative pressure means that the air pressure inside the laboratory is lower (i.e. negative) than the air pressure outside the laboratory. The laboratory is built with thick concrete walls and has a special sealing process, etc., so it has extremely high airtightness and prevents the air inside the laboratory from leaking outside. In addition, the ventilation system in the laboratory ensures that the air inside is constantly replaced with clean air, and the exhaust air from the laboratory is cleaned by a HEPA filter before being discharged. This prevents any pathogens, which may be contained in the air in the laboratory, from leaking out of the laboratory.
Q．Is there any risk that the virus could leak into the environment with wastewater from the BSL-4 laboratory?
A．At BSL-4 facilities, all wastewater from the laboratory area is first sterilized by high-pressure steam, and then chemically disinfected with a chemical agent. This process completely sterilize (kills) any viruses contained in the wastewater.
Q．Is there any risk that the virus will stick to the experimenter and leak outside?
A．When the experimenter leaves the laboratory, they decontaminate their laboratory suit (Positive Pressure Suits(PPS)) with a shower of disinfectant. At this stage, even if any viruses are on the suit, they will be sterilized (killed).
Q．Is there any risk of infected experimental animals escaping from the BSL-4 facility?
A．Experimental animals are kept in individually ventilated cages. Furthermore, it is necessary to pass through several rooms to get from the animal room to the exit of the laboratory area. The doors at the front and back of the several rooms are designed not to open at the same time. Thus, multiple systems are in place to prevent the escape of animals.
Q．You should have an emergency power supply in case of power outages due to natural disasters, shouldn’t you?
A．Emergency power supply has been installed in BSL-4 facilities in preparation for power outages due to natural disasters, including flooding. It has a capacity that can maintain the facilities' pathogen containment equipment normally. It is also redundant so that it can operate even if one unit fails.
Q．Is there any risk of the experimenter getting infected with the virus?
A．During experiments, the experimenter wears Positive Pressure Suits(PPS) (experimental suits), so they are completely isolated from the environment inside the laboratory. In addition, there are rules in place in the laboratory, such as not using sharp instruments as much as possible, to ensure the safety of the experimenters. Measures have been taken to ensure that the risk of infection to the experimenter is as low as possible, both in terms of equipment and in terms of work rules.
Q．What does “Positive Pressure” mean in the context of the Positive Pressure Suit (PPS)? Also, please explain how experimenters are protected by the PPS.
A．”Positive Pressure” means that the air pressure inside the Positive Pressure Suit (PPS) is higher (＝positive) than the air pressure outside the PPS. PPS has an airtight construction with no leaks and protects the experimenter from pathogens by physically isolating them from the laboratory environment. The positive pressure means that even if there is a leak at the surface of the PPS, the air in the laboratory will be pushed out so that it does not enter the inside of the PPS. This allows experimenters to carry out their activities without coming into contact with pathogens in the laboratory air.
Q．How heavy is the Positive Pressure Suit (PPS)?
A．The PPS weighs about 7 kg (varies depending on size).
Q．Why are Positive Pressure Suits (PPS) yellow?
A．In experiments, there are cases where you may handle culture fluid (red) or blood that contains viruses, etc. If any of these fluids containing viruses get on PPS, it is necessary to be able to see them easily. The color of PPS is yellow, as it is a color with relatively high contrast to the color of these fluids.
Q．What measures are being taken to minimize the possibility of human error?
A．Nagasaki University allows access to the BSL-4 laboratory to those who have fulfilled the conditions stipulated in the safety management regulations, such as by attending training courses. In addition, when conducting experiments in a BSL-4 laboratory, there are rules that require at least two people to be present at all times to check each other's actions, and that the experiment time is limited to six hours per day to prevent excessive fatigue. This ensures to prevent human error. Furthermore, we aim to prevent human error by preparing and regularly reviewing safe procedures for work in the laboratory, and by conducting regular training.
Q．What do they do if an experimenter feels unwell during the experiment ?
A．When entering the BSL-4 laboratory, multiple experimenters enter together to support each other's experiments. Staff constantly monitor the inside of the laboratory with surveillance cameras and other equipment. Thus, they can respond immediately when an experimenter feels unwell. The experimenters also conduct emergency response training in preparation for such cases. In addition, we are prepared for emergencies, such as conducting transport training in collaboration with Nagasaki University Hospital, which is a designated medical institution for class I infectious diseases within the same university. Then staff who enter and exit BSL-4 facilities are going to take AED training and cardiopulmonary resuscitation courses.
Q．How is access to and from the BSL-4 facility controlled?
A．Access to and from BSL-4 facilities is strictly controlled. The facility has a multi-layered personal authentication system, including security cards, and is also equipped with surveillance cameras and an access control system. Thus only authorized personnel are allowed to enter.
Q．Is there any risk that the viruses used in the experiments will be stolen or taken away?
A．Only those who have been registered and approved in advance are allowed to enter the building according to the rules for the BSL-4 facility. Furthermore, access to and from the BSL-4 laboratory is strictly controlled (as mentioned in the previous Q). Multiple researchers work at the same time, and they monitor each other when handling the virus. The situation inside and outside the laboratory is constantly monitored by surveillance cameras, etc. In addition, we plan to conduct regular training with the police in case of suspicious person, etc.
Q．What kind of information security measures are in place for the internet at the BSL-4 facility?
A．The network within the BSL-4 facility uses a closed network system that is separate from the internet and is only accessible within the building. In addition, there are strict restrictions on bringing electronic devices into the facility.