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Biosafety Levels and Their Meaning
Lab Academy
- Bioprocessing
- Cell Biology
- Centrifugation
- Safety & Biosafety
- Centrifuges & Rotors
- Essay
Working with biological material can sometimes pose a danger to laboratory staff and the environment. For this reason, staff must correctly handle material according established standards. These standards comprise four biosafety levels (BSLs) that typically apply to biological materials. Depending on the level, laboratory staff will have to possess a certain set of skills for handling potentially dangerous material. Aerosols forming during standard lab procedures like pipetting, mixing, and centrifuging poses the greatest potential risk for infection. To minimize the risk of infection from bioaerosols, primary and secondary barriers are used. A primary barrier could, for instance, be a biosafety cabinet (BSC). A secondary barrier might be an autoclave (BSL-2) or, in a higher-level situation, an installation inside a facility like a dedicated anteroom or ventilation system might serve as the barrier. Infectious material might include bacteria, viruses, cell cultures, parasites, or particular types of fungi. Depending on the security standard in place, special training may be required to handle these infectious materials. The laboratory supervisor is responsible for correctly educating laboratory staff.
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Biosafety level 1 (BSL-1)
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Examples of typical BSL-1 biological material
- Bacteria: Agrobacterium tumefaciens, Escherichia coli K-12, Pseudomonas fluorescens, Bacillus subtilis
- Viruses: canine adenovirus types 1 and 2, Bovine enterovirus
- Fungi: Saccharomyces cerevisiae
- Certain cell cultures> Endoparasites of animals and humans (e.g., some nematodes)
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Biosafety level 2 (BSL-2)
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Examples of typical BSL-2 biological material
- Bacteria: Chlamydia pneumoniae, Enterobacter spp., Mycoplasma pneumonia, Yersinia pseudotuberculosis
- Viruses: Herpes simplex virus, HIV (when performing routine diagnostic procedures or working with clinical specimens)
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Biosafety level 3 (BSL-3)
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Examples of typical BSL-3 biological material
- Bacteria: Yersinia pestis, Brucella abortus, Chlamydia psittaci, Pseudomonas mallei
- Viruses: West Nile fever, herpes B, hepatitis A
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Biosafety level 4 (BSL-4)
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Examples of typical BSL-4 biological material
- Ebola-Virus, Lassa-Virus, Marburg-Virus
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Requirements for standard laboratory equipment in BSL labs
Centrifugation
- Centrifuge vessels (aerosol-tight vessels required): decontamination after use required; as an alternative, sterile, single-use vessels may be used
- Aerosol-tight rotors
- Aerosol-tight caps that can be removed from the centrifuge for secure transport
- Use of vessels with high thermal, centrifugation, and chemical stability
- Imbalance sensors and emergency stop feature
- Detailed information in the operating manual about correct loading of the rotor
- Appropriate cleaning and disinfection protocols for rotors and rotor chambers provided by the manufacturer
- Clear manufacturer instructions for opening the centrifuge lid should power fail
Mixing
- Appropriate mixing technology used by the mixer to avoid splashes
- Use of robust, tight-closing, single-use vessels that can be autoclaved
- Use of vessels with high thermal and chemical stability
Incubation
- Regular filter checks
- No fan inside to avoid contamination
- Easy-to-clean chamber
- Copper chamber to avoid contamination
Pipetting
- Cleaning and disinfection procedures provided by the manufacturer
- Nonreactive pipettes that can be autoclaved
- Handling information provided by the manufacturer to avoid contaminations (air-cushion pipettes)
- Use of filter tips to avoid contamination of pipette interior
- Use of positive displacement tips to avoid contamination
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