Virtual Lab Manual

Biosafety Simulation

Synopsis
In the Biosafety simulation, you will be introduced to the concept of laboratory
containment, which is a very special type of laboratory required to work with hazardous
pathogens.

Laboratory containment
Containment in this context refers to the requirements set out by various bodies to safely
manipulate such pathogens. In this simulation, you will be able to experience a containment
level 3 (Biosafety level 3 or BSL3) research laboratory.

Learn about lab safety

You will become familiar with the basic protocols utilized to ensure safety, and you will be
presented with the specific basic safety rules for this type of laboratory (e.g. personal
protective equipment (PPE) and primary containment). In addition, you will learn how every
part of the laboratory is designed to avoid contamination. This includes learning about why
the pressure in the laboratory needs to be controlled at all times (secondary containment),
and how the microbiological safety cabinets are designed to offer protection).

Identify the hazard

In the Biosafety simulation you will be presented with a hypothetical case where you will
need to identify a potential bioterrorism agent that is classed as hazard group three
microorganism.

Please note that this simulation has been developed broadly around UK regulations. Some
protocols may differ depending on the specific regulations.

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Learning Objectives
At the end of this simulation, you will be able to…
● Understand how a Biosafety containment level III laboratory is constructed (e.g. air
flow and pressure control)
● Understand the basic safety rules of a Biosafety containment level III laboratory (e.g.
use of safety equipment)
● Handle microorganisms in a Biosafety containment level III laboratory
● Understand the concept of fumigation and how this is executed within a
microbiological safety cabinet.

Techniques in Lab
● Containment Level 3 good practice and aseptic technique
● Bacteria inoculation
● Microscopy

Theory
Anthrax
Anthrax, known as Bacillus anthracis is a Gram-positive, rod-shaped bacterium that is well
known for its use in past bioterrorism attacks, although it is naturally found in the soil.

Anthrax is classified as a Hazard group 3 pathogen due to its ability to cause disease and
potential to spread to the community. If left untreated, all types of anthrax can cause
death. However, effective prophylaxis or treatments are usually available.

Anthrax has been used as a bioweapon for its ability to form spores that are highly
resistant to heat, drying and common disinfectants such as 95% ethanol: the common
disinfectant of a Biosafety level II lab.

Figure 1: Gram stain of Bacillus anthracis.

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Anthrax spreads indirectly, via its spores, which can be picked up from the ground by
grazing animals. As a weapon, the spores are easily transported and can spread through
direct contact, and they are also light enough to spread through the air.

Bacillus anthracis can produce toxins that are very toxic to the human body, causing severe
human disease and ultimately death. Additionally, the presence of a polyglutamic capsule,
makes it more difficult for the immune system to fight it. Luckily, anthrax is very rarely
transmitted from person to person.

Laboratory biosafety
The principles of preventing unintentional exposure to pathogens and toxins, or their
accidental release determine laboratory biosafety. Laboratory biosafety levels are
categorized as follows:

Figure 1. Table with biosafety levels

GMP = Good microbiological practice. BSC = Biosafety cabinets

Biosafety level designations are based on lab design and practice required for working with
agents from the various hazard groups. In the UK Biosafety lab is referred to as a
Containment lab.

Setting up culture in BSL 3

Culturing in a Biosafety level III lab works in the same way as culturing in a normal lab.
However, more care is needed. For this reason, no flip top tubes are used and the “little
finger technique” is applied to open screw top lids safely when pipetting. Class III mode is
deemed safer when cultures exceed 25 mL, because the agent won't be so concentrated.

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