Biosafety And Biocontainment

The risk assessment is to be done by the owners / key stakeholders of the lab. Please refer to the answer to question 2 below which provides the details of how a risk assessment is to be carried out. Under Indian law, the Department of Biotechnology has clear guidelines on the implementation of the Biosafety rules and regulations.

Please refer to these links (link-1 & link-2) for further details.

Every country will have their own regulations and one needs to check the local regulations around the biosafety and follow the same. DBT in India provides the necessary certification/permission.

Risk assessment is a process and is a team effort. For performing a risk assessment in any lab practically, the team should necessarily comprise of the key stakeholder of the lab, engineering & maintenance engineer, safety person, microbiologist, and lab scientist or operator. The process as explained in the slide deck involves first the identification of the risks in terms of the organism in use, the processes to be followed, the potential routes of exposures, and the protocols to be followed in the lab. Once the identification is carried out a risk matrix is created to map each activity and identified risk against its severity and probability of occurrence. The matrix helps identity each activity into different categories of risk and hence the BSL level can then be assigned by the team. Once the BSL level is assigned, the team needs to work out the preventive measures for accidental exposure of the organism to the worker or environment and surroundings. Remember the hierarchy of control explained and apply the controls. Carry out a review with the team and identify the training requirements of the facility. Once this is done, you can get the risk assessment reviewed by an expert or the local regulatory body. You could also alternatively hire a consultant to do the risk assessment.

The purpose of biosafety is to prevent the biohazard from contaminating the person working with the biohazard and to protect the accidental release or exposure of the same to the environment and surroundings. The negative pressure in the BSC allows the air to flow away from the worker and is filtered while moving out of the BSC. Hence, the BSC needs to be under negative pressure.

This is based on the microorganisms being used in the labs and there cannot be a generalization on the same. A proper risk assessment needs to be carried out and then the Biosafety Level needs to be assigned. Generally, DHHs and Teaching hospitals would need a BSL – 1 or BSL – 2 category labs. If a DHH or a hospital is involved in research of exotic or higher risk category microbe then based on risk assessment, the same becomes applicable. For any lab, there is no generalization. Please carry out a thorough risk assessment and only then assign the Biosafety level.

In terms of decontamination, apart from decontaminating autoclaves, one could deactivate the material chemically by the use of a tested decontamination agent. Materials may also be properly bagged and sealed with surface decontamination of the bag and then sent for biomedical waste disposal through incineration.

This depends on the design of the lab. A proper cascade needs to be designed to achieve the correct negative pressure in the lab. This is a process to be taken up during facility design. There is no set level of negative pressure. However, good guidance would be to have a 10-15 KPa differential between adjacent rooms and progressively designed with a cascade.

This is dependent on a few factors:
i) Type of microorganism.
ii) Number of unit operations being performed.
iii) Number of people required to work in a lab.
iv) Area required for decontamination systems, drainage systems, HVAC systems, and exhaust systems.

Hence, there is no hard and fast rule around this. The aim should be to optimise the space to the minimum as possible without compromising on the safety and design of such labs.

The Biosafety cabinets need to comply to the requirements as specified in American Standard NSF 49; European Standard EN 12469; British Standard BS 5726; Australian Standard AS2252 and Japanese standard JIS K 3800.

There is no sufficient data on Corona Virus and its mutants and hence it is not possible to state whether BPE filters will work to contain SARS-CoV-2 Virus.

For a BSL 3 facility, we generally use the following types of BSC cabinets – Class II Type A1; Class II Type A2; Class II Type B1; Class II, Type B2 and Class III. Of these, Class II type A1 is near obsolete. For the Type A2, a thimble duct is permissible since the percentage of the exhaust is only 30% and there is 70% recirculation. In all other cases, hard duct is recommended since the control plenum is surrounded by a negative plenum and hence requires an external blower for suction to exhaust. A hard duct provides better integrity and also permits easier balancing of the airflow if a hard duct is used.

UV lamps are not recommended in BSC nor are they necessary. If installed, UV lamps must be cleaned weekly to remove any dust and dirt that may block the germicidal effectiveness of the ultraviolet light. The lamps should be checked weekly with a UV meter to ensure that the appropriate intensity of UV light is being emitted. UV lamps must be turned off when the room is occupied to protect eyes and skin from UV exposure, which can burn the cornea and cause skin cancer. If the cabinet has a sliding sash, close the sash when operating the UV lamp. Heat decontamination is not a must have but can be installed since heat decontamination is good methodology decontamination.

A manual fumigation method with proper procedural control is sufficient. Care will need to be taken to have all surfaces decontaminated and hence a good process control and monitoring are required. However, if one can afford a decontamination system through the HVAC system of the room, it is safer and consistent. Such systems can be automated.

Isolators and Bio Safety Cabinets are different concepts. The prime aim of a BSC cabinet is to protect the personnel and the environment with low importance given to product/sample safety. However, in case of the isolator, there is equal importance given to the product, personnel and environmental safety. The isolator is a word generally used for chemical entities while Biosafety is a concept used for biological entities. The Biosafety cabinet type III is based on a concept of negative pressure isolators. Hence, usage of a negative pressure isolator for BSL labs at level 4 and sometimes at level 3 can be acceptable. However, one needs to assess the costs involved. Isolators with positive pressure cannot be used in Biosafety labs.

Any electronic item in the lab needs to be compatible with the bio-sporicidal agent and standard disinfectants used in the lab. One could use wipes, or sprays to decontaminate the electronic item. The chemical compatibility of the electronic item to the proposed decontamination agents needs to be taken into consideration while ordering the equipment.

The question here is not clear. If the question is that do we apply clean room standards to a Biosafety lab. The answer is yes. Clean room design provides means of maintaining microbiological cleanliness in an area and it is good to apply the same to biosafety labs. This is generally applied to all microbiology labs in the industry.

Ideally BSL 4 labs should be isolated buildings with proper control on all entry and exit into the facility. While there are no specific guidelines on the neighbourhood or space requirements of the BSL 3 & 4 facilities, it is ideal to create sufficient buffer space equivalent to 1:1 or 2 around the lab area.

In India there are 4 facilities as available in the general domain:
i) High Security Animal Disease Laboratory (HSADL), Bhopal.
ii) Centre for Cellular and Molecular Biology, Hyderabad.
iii) All India Institute of Medical Sciences, New Delhi which has labs at BSL 1 to 4.
iv) Microbial Containment Complex at the National Institute of Virology (NIV) in Pune.

Fumigation using vaporized hydrogen peroxide is a safer and recommended method. The control of humidity, concentration, cycle time and temperature are key to a successful fumigation process

70% of Alcohol is a good germicidal agent for surface cleaning. Hydrogen peroxide is a safe agent for fumigation/decontamination of the room. As a general rule, it is a good practice to rotate the germicidal agents since it prevents any resistance by microorganisms to a given germicidal agent. The usage of commercially available germicidal agents with a rotational plan recommended by such suppliers is a good choice.

No

70% of Alcohol is a good germicidal agent for surface cleaning. Hydrogen peroxide is a safe agent for fumigation/decontamination of the room. As a general rule, it is a good practice to rotate the germicidal agents since it prevents any resistance by microorganisms to a given germicidal agent. The usage of commercially available germicidal agents with a rotational plan recommended by such suppliers is a good choice.

UV lamps are not recommended in BSC nor are they necessary. If installed, UV lamps must be cleaned weekly to remove any dust and dirt that may block the germicidal effectiveness of the ultraviolet light. The lamps should be checked weekly with a UV meter to ensure that the appropriate intensity of UV light is being emitted. UV lamps must be turned off when the room is occupied to protect eyes and skin from UV exposure, which can burn the cornea and cause skin cancer. If the cabinet has a sliding sash, close the sash when operating the UV lamp. Heat decontamination is not a must have but can be installed since heat decontamination is a good methodology decontamination.

There are consultants available who can help with the validation of the BSL 3 & BSL 4 laboratories. However, one has to bear in mind that the owners of the laboratory are responsible primarily for the same. The validation needs to be done in accordance with the guidelines specified by the local authority (either under department of health and safety or if available department of biotechnology), WHO, CDC, BMBL and NIH.

Please find the links to the guidelines from WHO, CDC for your reference.

Within the specified guidelines, except for the autoclaves used for decontamination or kill (sterilization) no SAL limits are specified. The BSL III labs are generally graded D or C based on risk assessment and the cleanliness required + application for which it is being used and hence one needs to follow the clean room guidelines specified for microbiology lab under the FDA.