Biosafety levels

A biosafety level is the level of the biocontainment precautions required to

isolate dangerous biological agents in an enclosed facility. The levels of
containment range from the lowest biosafety level 1 to the highest at level 4.
The term "containment" is used in describing safe methods for managing
infectious materials in the laboratory environment where they are being
handled or maintained. The purpose of containment is to reduce or eliminate
exposure of laboratory users, other people, and the outside environment to
potentially hazardous agents. Biocontainment can be classified by the relative
danger to the surrounding environment as biological safety levels (BSL). As of
2006, there are four safety levels. These are called BSL1 through BSL4.
Biosafety level 1 (BSL-1)
BSL-1 is the lowest security level for handling biological material. This kind of
material poses no or only a low risk to healthy adult humans and presents
minimal potential hazard to laboratory personnel and the environment. BSL-1
laboratories do not have to be separated from the rest of a building. Laboratory
workers can do their work on open bench tops, and there is no need to use
special equipment. Standard microbiology practices usually suffice to protect
laboratory workers and other employees in the building. This means, for
example, that no mouth pipetting is allowed and splashes and aerosol
formation should be avoided. Spills have to be cleaned up immediately and all
work surfaces should be decontaminated each time work is finished. Eating,
drinking, and smoking are not allowed in the lab. To protect themselves,
workers generally wear eye protection, gloves, and a lab coat. It is
recommended that access to the laboratory be limited. In general, an autoclave
is only required when working with genetically modified organisms or modified
genetic elements (e.g., plasmid vectors). Biohazard signs are posted whenever
infectious agents are present. 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)
Safety protocols for biosafety level 1 labs — which require only standard
microbial practices — include:
 Mechanical pipetting (no mouth pipetting allowed)
 Safe sharps handling
 Avoidance of splashes or aerosols
 Daily decontamination of all work surfaces when work is complete
 Regular handwashing
 Prohibition of food, drink, and smoking materials
 The use of personal protective equipment (PPE), such as goggles, gloves,
and a lab coat or gown
 Biohazard signs
BSL-1 labs also require immediate decontamination after spills. Infectious
materials should also be decontaminated prior to disposal, generally through
the use of an autoclave.
Biosafety level 2 (BSL-2)
All activities in a BSL-2 laboratory require higher security standards than in a
BSL-1 laboratory. The biological material used in a BSL-2 laboratory consists of
bacteria, viruses, and organisms associated with human diseases. The potential
pathogenic or infectious organisms subject to BSL-2 standards pose a moderate
hazard to healthy adult humans. However, because of their potential to cause
human disease, great care must be taken to prevent percutaneous injury, and
employees are subject to regular medical surveillance if necessary. The main
difference in the work procedures followed in a BSL-1 laboratory and a BSL-2
laboratory is that employees in a BSL-2 laboratory will use a BSC as a primary
barrier for potentially hazardous aerosols. These aerosols are generally formed
during standard work procedures like mixing, pipetting, and centrifuging, so
these activities should be carried out under a BSC. At a minimum, an autoclave
has to be present in the lab as a second barrier. Windows open to the outside
are not recommended. A sink for washing hands must be placed in direct
proximity to the exit door. The laboratory should be easy to clean and
decontaminate. Inside the laboratory, special lab coats, gloves, and face and
eye protection should be worn. Special precaution must be taken with sharps
that can cause injury. Access to BSL-2 laboratories must be restricted.
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)
In addition to the safety protocols established for BSL-1 labs, BSL-2 labs are
subject to the following safety controls:
 The use of PPE, including lab coats, gloves, eye protection, and — in
some cases — face shields
 All procedures that could cause infection from aerosols or splashes must
be performed within a biological safety cabinet.
 Decontamination of infectious materials prior to disposal, generally
through the use of an autoclave
 Self-closing, lockable doors
 Access to a sink and eyewash station
 Biohazard warning signs
Access to a biosafety level 2 lab is far more restrictive than to a biosafety level
1 lab. Outside personnel, or those with an increased risk of contamination, are
often restricted from entering the area while work is underway.
Biosafety level 3 (BSL-3)
BSL-3 involves handling indigenous or exotic agents that may cause serious or
potentially lethal diseases through inhalation and/or pose a serious threat to
the environment. Compared with the BSL-2 lab, BSL-3 labs require additional
security precautions: Only limited access to the laboratory is allowed.
Moreover, laboratory personnel remain under medical surveillance and are
offered appropriate immunizations for the agents handled or potentially
present in the laboratory. All laboratory waste has to be disinfected, and the
clothes worn in the lab must be decontaminated before laundering. Walls,
ceilings, and windows must be sealed, and floors must be nonskid and resistant
to chemicals. The design, operational parameters, and procedures of a BSL-3
facility must be verified and documented prior to the start of operations.
Facilities must be documented and verified again a minimum of once a year. In
terms of a primary barrier, all open manipulations of biological material must
be done under a BSC (preferably Class II or Class III) or in a closed containment
system, and special protective clothing must be worn. BSL-3 laboratories must
have separate exits with self-closing doors as the second barrier. Moreover, an
air-ventilation system providing negative air flow is required that does not
recirculate exhaust.
Examples of typical BSL-3 biological material
 Bacteria: Yersinia pestis, Brucella abortus, Chlamydia psittaci,
Pseudomonas mallei
 Viruses: West Nile fever, herpes B, hepatitis A
Common safety controls within a BSL-3 lab include:
 The use of PPE, including goggles and gloves; respirators may also be
required
 The use of solid-front wraparound gowns, scrub suits, and/or coveralls is
often required
 Access to a hands-free sink and eyewash station available near the exit
 Sustained directional airflow to draw air into the laboratory from clean
areas toward potentially contaminated areas (exhaust air cannot be
recirculated)
 Self-closing set of locking doors with access away from general building
corridors
 Access to a BSL-3 laboratory is restricted and controlled at all times.
Biosafety level 4 (BSL-4)
BSL-4 entails handling very dangerous and exotic biological material that may
cause infection inside the laboratory as the result of aerosols. Other agents can
have a very similar antigenic relationship to existing BSL-4 material, but there is
insufficient data available to designate the level for these agents. Biological
material also handled at this level relates to existing infectious material where
the disease transmission is unknown. In addition to the precautions taken at
the BSL-3 level, BSL-4 requires additional protection: All clothes must be
changed before entering the lab, and employees must shower before leaving.
In terms of primary barriers, all work has to be carried out in a Class III BSC or
in a Class I or II BSC in combination with the lab worker wearing a full body, air-
supplied positive pressure suit. A BSL-4 laboratory is usually located in a
separate building or in a specific insolated area of a building; this isolation
serves as a second barrier. This level also requires dedicated supply and
exhaust ventilation equipment with vacuum and decontamination systems.
Depending on the material and requirements, special installation might also be
necessary.
Examples of typical BSL-4 biological material
 Ebola-Virus, Lassa-Virus, Marburg-Virus
In addition to biosafety level 3 considerations, biosafety level 4 laboratories
must follow these safety protocols:
 Personnel must change clothing before entering the facility and shower
upon exiting
 All materials must be decontaminated before leaving the facility
 Personnel must wear the PPE from lower BSL levels, as well as a full-
body, air-supplied, positive pressure suit
 Access to a Class III biological safety cabinet
BSL-4 labs are extremely isolated, often located in an isolated and restricted
zone of a building or in a separate building entirely. BSL-4 labs also feature a
dedicated supply of exhaust air, as well as vacuum lines and decontamination
systems.

Lab safety and good laboratory practice (GLP)

GLP (Good Laboratory Practices) is generally defined in regulating laws from
various jurisdictions as: “a set of rules and criteria for a quality system
concerned with the organizational process and the conditions under which
non-clinical health and environmental safety studies are planned, performed,
monitored, recorded, reported and archived.”
Good lab practices are an integral part of conducting research safely.
Engineering controls can limit exposure to hazards and PPE can protect a
researcher's body, but making sure your behavior doesn't expose you or your
colleagues to risks is important. These safe lab practices cover many of the
common routes of exposure but are by no means a comprehensive list.
Dress code safety rules
As you’d expect, laboratory dress codes set a clear policy for the clothing
employees should avoid wearing in order to prevent accidents or injuries in the
lab. For example, skirts and shorts might be nice for enjoying the warm
weather outside, but quickly become a liability in the lab where skin can be
exposed to heat or dangerous chemicals.
 Always tie back hair that is chin-length or longer.
 Make sure that loose clothing or dangling jewelry is secured, or avoid
wearing it in the first place.
 Never wear sandals or other open-toed shoes in the lab. Footwear
should always cover the foot completely.
 Never wear shorts or skirts in the lab.
 When working with Bunsen burners, lighted splints, matches, etc., acrylic
nails are not allowed.
No Food or Drink
 Consuming food in the lab can pose many hazards.
 Eating or drinking in the lab can, first and foremost, increase your risk of
exposure to hazardous materials.
 Food or drink can leave a mess increasing the risks for contamination of
your experiments and potentially attracting pests.
 Eating or drinking in the lab can also be a distraction that can lead to a
spill or more serious incident.
Wear Your PPE and Proper Lab Attire
  Lab coat, gloves, eye protection, and appropriate attire should be worn
at all times in the lab.
 Long pants and shoes completely covering the top of the foot should be
worn at all times when working in the lab.
 Lab coats will protect your clothes and your skin from splashes, spills, or
other exposures to chemical or biological agents, and flames in some
cases.
 Safety glasses or goggles will protect your eyes from physical of chemical
harm. Skin will heal after minor burns or lacerations but your eyes will
not. Eyes are fragile and safety glasses take about three seconds to put
on, an eye injury can be permanent.
 Gloves protect your skin from hazardous materials your hands may come
into contact with. However, exposure can occur when removing gloves
and disposing of them. Follow the steps in the video below to properly
remove any gloves used in the lab.
Good Hygiene
 Wash hands after handling any hazardous materials, before and after
eating, and before leaving the lab.
 Keeping personal items separate from lab work. This will prevent spread
of hazardous reagents and cut off a potential exposure route.
 Do not apply cosmetics while in the lab. Applying anything to your face,
especially around your mouth or eyes, pose a significant risk of exposure.
 Dry and cracked skin can provide a route to exposure. Using lotion to
keep the skin on your hands healthy can help prevent exposure.
Use Proper Storage Containers
 This applies to individual containers, storage cabinets, and waste.
 Storing organic solvents in plastic bottles can compromise the container,
just like acids in metal containers or HF in glass. Chemicals should be
stored in containers made of materials that will not react.
 Large volumes of flammable chemicals must be stored in fire rated
cabinets. Acids and caustics should ideally be stored in separate cabinets
lined with plastic to prevent any vapours from reacting with the metal
housing. Chemicals known to react violently when mixed should be
stored separately.
  As with chemical storage, waste should be stored in non-reactive
containers, or containers with non-reactive liners.
Label Your Work Space
 All containers should be labelled with their contents. This is crucial so
those working near you and anyone visiting the lab will know what
hazards may be present. Ideally the hazards present should be included
on any label.
 Any research process with a particular hazard should also be labelled
with that hazard.
Participate in Safety Exercises
 Ensure all lab members are familiar with the lab's safety equipment.
 Make sure everyone knows where the nearest fire pull station,
extinguisher, spill kit, first aid kit etc.
 Know how to get out of your building and where to go after and
evacuation.