Laboratory biosafety means the protection of people and environment from exposure to contagious biological material generated in the laboratory. Laboratory biosafety is part of the laboratory safety work. Issues regarding laboratory biosafety can be put to the department's contact person for laboratory biosafety, or to the biosafety administrator at KI.
Rules and directions
The specific regulations that, above all, control the work on laboratory biosafety in Sweden, are included in the Swedish Work Environment Authority's provisions regarding:
- Microbiological Work Environment Risks– Infection, Toxigenic effect, Hypersensitivity
- Contained Use of Genetically modified Micro-organisms
You will find links to these provisions under "Acts and Provisions - work environment and health".
There is no Swedish book to be used for more detailed information within the subject of laboratory biosafety, however, the WHO has published a manual in English.
Definitions - Laboratory biosafety
Laboratory biosafety: the protection of people and environment from exposure to contagious biological material generated in the laboratory. Further information can be found under Biosecurity.
Micro-organism: A micro-organism is defined as "every microbiological unit, cellular or non-cellular, which is able to reproduce, or transfer genetic material". They include, but are not limited to, bacteria, viruses, protozoa and cell cultures of higher organisms. Note that cell cultures from higher organisms in this context are considered micro-organisms, but that tissue/multicellular organisms are not covered by the regulations.
Risk group: Biological agents are divided into four groups, where a micro-organism is classified as a low risk if it is unlikely to cause severe infection in humans, and the possible resulting infections will be mild. A list of the classification of specific micro-organisms can be found in the Provision "Microbiological Work Environment Risks– Infection, Toxigenic effect, Hypersensitivity" (link).
Note that the classification of the micro-organism is not related to the severity of the consequences a possible contamination would have in society.
Containment level: Laboratories are divided into four containment levels, depending on which protective measures they contain. A list of the specific protective measures required for the various containment levels can be found in the provision regarding Microbiological work environment risks. The various containment levels have been adapted for the use of micro-organisms with similar risks (refer to risk group definition).
Also refer to "Permits and reports for handling micro-organisms".
GMM: GMM is defined as "a micro-organism, in which the genetic material has been altered in a manner that would not occur naturally through mating or natural recombination", in accordance with Section 2, Ordinance SFS 2000:271, interpreted in the provision "Contained Use of Genetically modified Micro-organisms".
Note that tissue/multicellular organisms are not considered to be micro-organisms, and are instead covered by the regulations for genetically modified organisms (GMOs). The gene modification must not necessarily have been integrated in the genome, or constructed in the user's own activities. Also refer to "Permits and reports for GMMs".
Blood infection: It is primarily Hepatitis B and C as well as HIV that can be transmitted through human blood handling. Note that blood infection is not only transmitted through blood handling, but also through material that has been contaminated by blood, such as tissue, exudate, spinal fluid, urine, pipette tips etc. Cell cultures may also transmit blood infections, refer to "Cell cultures and blood handling".
Laboratory biosecurity: describes how the handling and storage of micro-organisms should be carried out, in order that they cannot intentionally be spread and thereby cause harm to human beings or environment.
Risk assessments Biosafety
The Swedish Work Environment Authority's provisions regarding Microbiological work environment risks (look under "Acts and provisions – work environment and health") describe how, and when, to make a documented risk assessment for micro-organisms which have not been genetically modified. For genetically modified organisms (GMM), a risk assessment must always be carried out.
Further information on page "Registration, permits and risk assessment for GMM". The risk assessment shall be carried out in order that everyone shall have a safe work environment, and to raise risk awareness.
Micro-organisms that cause infectious diseases in humans are classified, in accordance with increasing hazardousness, into risk groups 2-4. The ones that do not cause infectious diseases are classified as risk group 1. They may still cause ill-health due to hypersensitivity or toxigenic effects. Examples of micro-organisms that do not normally cause infectious diseases are cell cultures, as well as E. coli, which has been adapted to the laboratory (i.e., not pathogenic).
In appendix 2 B to the provision "Microbiological work environment risks", there is a list of bacteria, viruses, fungi and parasites, classified in risk groups 2-4, to be used as a guideline.
A risk assessment must always be made for microorganisms in risk groups 2-4. A risk assessment might be necessary for micro-organisms in risk group 1. It is up to the user to evaluate whether a micro-organism in risk group 1 is in need of a more thorough written risk assessment.
The Biosafety Committee at KI has created a risk assessment form for microorganisms, called BARA (Biological Agents Risk Assessment). The form can be used to assess the risk of handling micro-organisms (including cell cultures). Please note that an extra form for handling microorganisms in animal experiments, which is called "B2 Risk Assessment for Animal Handling" is inlcuded in BARA and may be used for this type of work.
For risk assessment of blood and other huaman sample material, the Biosafety Committee has created a form called HUMRA (Human Sample Material Risk Assessment).
Note that the BARA form should not be used for microorganisms that have been genetically modified, refer instead to "GMM-applications".
Notification/permit is required for all microorganisms in risk group 2 and higher. Also refer to "Applications to handle microorganisms".
When assessing the risk of chemicals, refer instead to "Risk assessment – Chemicals".
Protective measures Biosafety
The protective measures that should be chosen for the handling of biological material depends on the manner in which the material transmits infection. Often, chemical and biological material are handled simultaneously, which must be taken into consideration. The choice of protective measure shall be defined during the risk assessment. The risk assessment should be made on a form, BARA when handling biological material.
Further information can be found under "Risk assessment".
The risk assessment for the experiment shall state whether gloves need to be used, and if so, which type. When choosing gloves for microbiological work, both penetration ability and resistance against cuts shall be taken into consideration. For biological material, there are three classes of gloves. Gloves in the three classes may consist of different thickness and different material, which have differing degrees of resistance to chemicals and puncturing.
Your hands shall always be washed and disinfected upon taking off the gloves after microbiological work. The disinfectant destroys the protective properties of the glove. Remember that the gloves become infectious, change them often. Note that not all microbiological work necessarily must be carried out wearing gloves.
The examination gloves are in a box marked CE, and most often the code EN455. These have been developed for patient protection. They have not been tested by an independent source. On the other hand, the producer guarantees that only a certain percentage of the gloves have holes in them. If the box reads AQL 1.5, it means that the producer has tested that 1.5 per cent, or less, of the gloves have holes in them. This type of glove, therefore, gives a false sense of security in the laboratory. It does not claim to protect against chemicals. One example is the Evercare from SelfaTrade.
Gloves for individual protection
The box with the gloves (or alternatively, the actual glove, if they are packaged individually) to be used for your personal protection, is marked with the code EN374. These are sub-grouped from one to three. EN374-1 is very similar to an examination glove, but may be used when handling milder chemicals, such as detergents. For the handling chemicals and micro-organisms, in which a higher level of protection is required, you often need gloves marked EN374-2 or EN374-3. These also have symbols with biological hazard/E-flask/beaker with a question mark.
The ones with a biological hazard symbol have been tested with water leakage tests (i.e., not with micro-organisms), the ones with a E-flask/beaker with a question mark, have been tested for various chemicals, and have passed the test for some of them. Note that different glove materials are resistant to different chemicals, refer to "Protective measures - Chemical Safety" for an explanation of which gloves are suitable for what chemicals.
Gloves marked EN374-2 /-3 have been tested by independent organisations. Examples of gloves marked EN374-2, as well as the biological hazard symbol, are the Touch N Tuff nitrile gloves.
Gloves that have been tested to protect against micro-organisms
Only the gloves marked with ASTM F1671-97b (American standards, for which there is no European equivalent), have been tested for penetration by micro-organisms. One example of gloves with this marking is Eco Shield, from the SHIELD company.
When carrying out microbiological work at KI, protective clothing, such as a laboratory coat, shall always be worn. The clothing shall be removed when leaving the work area. When transporting micro-organisms, e.g., within the department, but accessing public areas, micro-organisms shall be stored in a transport container or similar (refer to "Transport of dangerous goods and biological specimens"), and protective clothing shall be removed.
Ventilated work areas
The risk assessment for the experiment shall state whether ventilated work areas need to be used, and if so, which type. It is important to differ between microbiological safety cabinets (also known as biological safety cabinets) and ventilated work areas adapted to handling chemicals. A microbiological safety cabinet is adapted for working with micro-organisms. Among other things, it has an HEPA filter, in order to purify the outgoing air. The majority of the air circulates, most often, but not always, inside the microbiological safety cabinet, and HEPA filtered air blows from the inner ceiling of the microbiological safety cabinet (this is called a class 2 microbiological safety cabinet).
A ventilated work station adapted to handling chemicals (e.g., downflow bench, local extraction system and fume cupboards) have no HEPA filter, but may have, e.g., a coal filter, which does not protect against micro-organisms in the air. Ventilated work stations adapted to handling chemicals do not, generally, protect the environment against micro-organisms. It is particularly important not to use a ventilated work station that only provides product protection, where the HEPA filtered air is going from the material to the laboratory assistant. This can be tested by holding a piece of paper over the work surface, and see which way the paper moves.
It is also important to consider whether the ventilated work station is connected to the exhaust (the ventilation of the building) or not. A microbiological safety cabinet is often disconnected from the exhaust, and such a bench does not protect against chemicals, neither through, nor throught the air exhaust wich is instead recirculated in the room.
Further information on the safety posts can be found in the Swedish Work Environment Authority's memo on microbiological safety posts.
Monitoring ventilated work stations at KI
Ventilated work stations at KI that are connected to the ventilation of the building ("exhaust-connected"), are checked annually by Akademiska hus, with regard to the air volume and alarms. Akademiska hus does not monitor the ones that are not exhaust-connected on a regular basis. At the checks arranged by Akademiska hus, the HEPA filters are not checked. HEPA filters may be destroyed through exposure to chemicals.
Other methods to protect against infection
When the only requirement is a splash guard, which is often the case for blood handling, no additional ventilation is needed. In these cases, a piece of Plexiglas, visor or goggles, as well as mouth protection shall be used. The risk assessment determines which type of protection to use. In these situations, a fume cupboard is often a good method of protection.
When working with specific micro-organisms, for which there is a vaccine, as well as when doing work that entails a risk of blood infection, the laboratory assistant shall be offered vaccination, upon risk assessment/medical evaluation. Read more in the page "vaccination".
In the event of a spill of biological material, it is important to quickly take measures, in order that no harm comes to staff or persons in the surroundings:
1. Seal off the area. This should be done to avoid exposure to aerosols.
2. Read the risk assessment.
3. Contact any specialists for advice, such as the occupational health service, infection clinic or the biosafety administrator.
4. Decontaminate, if possible. Never let the cleaning staff decontaminate! They do not have the expertise required.
The risk assessment regulates how to decontaminate. It takes approx. 5-10 minutes for all the aerosols to settle. Most often, the area should be covered with paper towels or equivalent material, after which a disinfectant chemical shall be poured onto the paper towels. It is important that the chemical has a sufficient contact time, i.e., 5-10 minutes. Thereafter, the material shall be handled as contagious. The surface should be disinfected again, and then washed with a non-oxidizing chemical, such as ethanol or detergent.
Note that the disinfected chemical shall be proven effective against the biological material in question.
In the event of a spill on a microbiological safety cabinet, item 1 can normally be skipped. In the event of a contamination of a centrifuge or incubator, the lid/incubator door shall instead be closed, in order for any aerosols to settle before it is cleaned in accordance with items 2-4 above.
Blood has a buffering ability, and therefore, ethanol is not sufficient in the event of a spill, and Virkon, hypochlorite or equivalent shall be used. Any incident shall be reported as an anomaly, as well as to the immediate manager. If a person has been exposed, it is often not enough to report the incident as an anomaly. In such a case, refer to "Reporting incidents" .
In the event of a chemical spill, refer instead to.....
Handling of samples with possible risk of microorganisms in risk group 4
Due to the ongoing Ebola outbreak, KI’s Biosafety committee would like to clarify that no samples suspected to contain risk group 4 microorganisms should be handled at Karolinska Institutet’s premises.
Samples from geographical regions with active outbreaks of risk group 4 pathogens should be avoided unless all risks of infection can be excluded. If there is any suspicion of risk group 4 microorganism in a sample, such a sample shall directly be safely transported to the Public Health Agency of Sweden. KI’s Biosafety committee refers to the guidelines from Karolinska University Laboratory (KUL) regarding how to manage a sample with suspected or confirmed contamination of Ebola or other viral hemorrhagic fever.
About transport to the Public Health Agency (pdf, in Swedish)
- Biosafety delegates 2018 (pdf, 136.85 KB)
- Risk assessment form BARA incl. animal handling (docx, 64.94 KB)
- Risk assessment form HUMRA (docx, 61.86 KB)
- Hantering på KUL av prov med Ebolamisstanke eller annan viral hemorrhagsisk feber (pdf, 219.37 KB)
- Rollbeskrivning för institutionernas biosäkerhetsombud och delegering (pdf, 177.23 KB)
- Anvisningar vid icke hanterade avvikelser och incidenter inom miljö, arbetsmiljö och säkerhet (pdf, 190.63 KB)
- KI blood rules and other human sample materials (pdf, 395.23 KB)