CENTRO ESCOLAR UNIVERSITY

SCHOOL OF MEDICAL TECHNOLOGY

MOLECULAR BIOLOGY
LABORATORY

NAME: ____________________________ DATE: ______________

YR/SECTION: ______________________ GROUP: ____________
ACTIVITY # 2
BIOSAFETY AND PPE

LEARNING OUTCOMES
1. Recognize distinctions between Biosafety Levels (BSL) identify PPE used for working in the
lab under BSL classifications
2. Demonstrate and assess the proper use of Personal Protective Equipment (PPE)

PRE LAB-DISCUSSION
Biosafety is the application of safety precautions that reduce a scientists’ risk of exposure to a
potentially infectious microbe and limit contamination of the work environment.

Biosafety levels (BSL), which provides minimum standards for safe handling of microbes at
each level. BSLs are defined and containment practices are detailed by the Centers for Disease
Control and Prevention (CDC) for laboratories in the United States.
We designate most of our labs under 4 special hazard categories called biosafety levels (BSLs).
Each level has specific controls for containment of microbes and biological agents. Each
biosafety level builds on the controls of the level before it.
All of these levels follow “standard microbiological practices”: which are those practices
common to all labs, which include not eating, drinking, or applying cosmetics, washing hands
after working in the lab, routinely decontaminating work area.

TYPES OF BSL LEVELS

BSL1: If you work in a lab that is designated a BSL-1, the microbes there are not known to
consistently cause disease in healthy adults and present minimal potential hazard to
laboratorians and the environment.
An example of a microbe that is typically worked with at a BSL-1 is a nonpathogenic strain of E.
coli.
 CENTRO ESCOLAR UNIVERSITY
SCHOOL OF MEDICAL TECHNOLOGY
MOLECULAR BIOLOGY
LABORATORY

Work can be done on an open lab bench, requires a sink. Requires personal protective
equipment (PPE) such as lab coats, gloves, eye protection.

BSL 2: BSL-2 builds upon BSL-1. If you work in a lab that is designated a BSL-2, the microbes
there pose moderate hazards to laboratorians and the environment.
The microbes are typically indigenous and associated with diseases of varying severity. An
example of a microbe that is typically worked with at a BSL-2 laboratory is Staphylococcus
aureus.
It includes various bacteria and viruses that cause only mild disease to humans, or are difficult
to contract via aerosols in a lab setting, such as Clostridium difficile, most Chlamydiae, hepatitis
A, B, and C, influenza A viruses, Salmonella.
BSL-2 differs from BSL-1 in that: laboratory personnel have specific training in handling
pathogenic agents and are directed by scientists with advanced training; access to the
laboratory is limited when work is being conducted; extreme precautions are taken with
contaminated sharp items; and certain procedures in which infectious aerosols or splashes may
be created are conducted in biological safety cabinets or other physical containment equipment.

BSL-3: is required for work involving indigenous or exotic agents, and they can cause serious or
potentially lethal disease that are transmitted through the air (via aerosols).
Respiratory transmission is the inhalation route of exposure. Laboratory personnel must receive
specific training in handling pathogenic and potentially lethal agents, and must be supervised by
scientists competent in handling infectious agents and associated procedures.
Lab personnel are under medical surveillance and might receive immunizations for microbes
they work with. All procedures involving the manipulation of infectious materials must be
conducted within Biosafety Cabinets (BSCs) or other physical containment devices, or by
personnel wearing appropriate personal protective equipment (e.g respirators).
A BSL-3 laboratory has special engineering and design features that prevent the release of
microorganisms to the environment. Facilities have hands free sink, exhaust air cannot be
recirculated, entrance is through two sets of self-closing and clocking doors.
Microbes that are worked on in BSL3 facilities are Mycobacterium tuberculosis which cause
tuberculosis, etc.
 CENTRO ESCOLAR UNIVERSITY
SCHOOL OF MEDICAL TECHNOLOGY
MOLECULAR BIOLOGY
LABORATORY

BSL-4 labs build on the containment requirements of BSL-3 and is the highest level of biological
safety. BSL-4 labs are required for work with dangerous and exotic agents that pose a high
individual risk of life-threatening disease, aerosol transmission or unknown risk of transmission.
The microbes in BSL4 labs cause infections that are frequently fatal and generally there are no
vaccines or treatments for these infections. There are only a small number of such labs in the
U.S (<10) and the world.
Laboratory staff must have specific and thorough training in handling extremely hazardous
infectious agents. Access to the laboratory is controlled by the laboratory supervisor. All
handling of agents must be performed in a gas tight Class III Biosafety Cabinet or by personnel
wearing a positive pressure protective suit.
BSL-4 Laboratories have special engineering and design features to prevent microorganisms
from being released into the environment. The lab is in a separate building or isolated restricted
zone of the building, and has a dedicated supply and exhaust air, as well as vacuum lines and
decontamination systems.
Personnel mush change clothing before entering and shower upon exiting. Most of the
pathogens worked on are viruses: Crimean-Congo hemorrhagic fever caused by Ebola, Junin,
Lassa, Machupo, Marburg viruses , and tick-borne encephalitis virus complex (including
Absettarov, Hanzalova, Hypr, Kumlinge, Kyasanur Forest disease, Omsk hemorrhagic fever,
and Russian Spring-Summer encephalitis).
 CENTRO ESCOLAR UNIVERSITY
SCHOOL OF MEDICAL TECHNOLOGY
MOLECULAR BIOLOGY
LABORATORY

Personal Protective Equipment (PPE)

Personal Protective Equipment, or PPE, is the clothing and equipment that forms the last line of
defense between you and harmful materials in the laboratory environment. It’s essential that you
know what you should be wearing, when you should be wearing it, and how it should be stored,
cleaned, maintained and disposed of.

Basic PPE provided in the laboratory includes: Disposable gloves, lab coat, safety glasses.

Disposable gloves
serve as a barrier between your hands and any chemical, biological, or physical hazards that
can enter your body through your skin. These gloves cannot protect you from all barriers, and
most disposable gloves we use in a lab are made of some synthetic material, such as nitrile or
latex.
Any prolonged exposure to a chemical agent can permeate these types of gloves, so it is critical
that you safely remove your gloves if you are exposed to a chemical spill and it comes in
contact with your gloves.
Gloves should be changed when they are contaminated by biological or chemical hazards.
There are many other types of gloves, such as ones for handling very hot items out of the
autoclave, or for very cold items out of a -80degC freezer or for handling liquid nitrogen. The
type of glove you use depends on the work you are doing in the lab.

Lab coats
protect the user's skin and personal clothing from accidental contact with biological or chemical
hazards. They also prevent the spread of contamination outside of the lab (provided they are
not worn outside the lab).
Lab coats serve as a removable barrier in the event of a spill or splash of hazardous
substances. They should fit the user well (not too large or it will get caught on things and not too
small as it may hinder movement), be long-sleeved, a secure cuff, knee-length or longer, fire
resistant material, high buttons to provide exposure of the chest or neck area.
They come in a variety of materials and provide varying degrees of protection. There are splash
resistant coats, static free coats, chemical resistant coats and flame-resistant coats.
Most lab coats we use in a teaching lab are made of a synthetic material that is fire resistant.
Always select a coat that provides the type of protection that is appropriate for your needs.
 CENTRO ESCOLAR UNIVERSITY
SCHOOL OF MEDICAL TECHNOLOGY
MOLECULAR BIOLOGY
LABORATORY

Safety glasses/goggles
provide a barrier to your eyes and prevent exposure to chemical (eg chemical reagents)
physical (eg dust, flying objects), and biological (eg bacterial culture splashes) hazards.
Most microbiology labs have standard safety glasses or goggles that wrap around the eyes and
avoid splashes, they may have features such as UV light barriers, impact features, and some
are designed to be worn over your regular glasses, these are called "Over the glasses" (OTG)
safety glasses.
There are also labs that have high intensity light sources such as UV light, lasers and the
appropriate safety glasses must be used.

Face shields
should be worn whenever the entire face needs protection. Such as when there is a potential
that an aerosol of chemical or biological hazardous material may be created or whenever
chemical or biohazards could splatter, or whenever there is the potential for flying particles or
sparks.
A face shield should always be worn whenever handling tissue samples or animals where there
is the potential for infectious transmission. Safety glasses or goggles should always be worn
underneath a face shield for maximal protection.

Masks
that are disposable may need to be worn, especially during the COVID-19 pandemic and will
limit the transmission of infectious agents. Disposable masks come in a variety of materials and
levels of protection.
Typical facemasks are loose-fitting, disposable masks that cover the nose and mouth, such as
surgical masks and nuisance dust masks. Facemasks are not approved by the National Institute
for Occupational Safety and Health (NIOSH) for protection against any regulated hazardous
material.

Facemasks help stop droplets from being spread by the person wearing them. They also keep
splashes or sprays from reaching the mouth and nose of the person wearing the facemask and
are therefore useful when cleaning up spills of infectious materials.
 CENTRO ESCOLAR UNIVERSITY
SCHOOL OF MEDICAL TECHNOLOGY
MOLECULAR BIOLOGY
LABORATORY

They are not designed to protect you against breathing in gases, vapors, or very small particles.
Facemasks should be used once and then disposed of.
There are different types of masks used for different purposes, specialty disposable face masks
such as the N95 or KN95 masks are respirators which are approved by NIOSH for use against
certain selected airborne particulates when used as part of a respiratory protection program.

PROPER DONNING & DOFFING OF PPE

1. BSL - 1 & BSL – 2

DONNING
: LABORATORY GOWN
: MASK OR RESPIRATOR
: Goggle or Face shield
: Gloves
DOFFING
: GLOVES
: GOGGLES OR FACE SHIELD
: GOWN
: MASK OR RESPIRATORY

2. BSL – 3
DONNING
: SHOE COVER
: LAB GOWN
: RESPIRATOR
: FACE SHIELD
: HAIR CAP
 CENTRO ESCOLAR UNIVERSITY
SCHOOL OF MEDICAL TECHNOLOGY
MOLECULAR BIOLOGY
LABORATORY

: INNER GLOVES
: OUTER GLOVES

DOFFING
: OUTER GLOVES
: FACE SHIELD
: HAIR CAP
: LAB GOWN
: RESPIRATOR
: SHOE COVER
: INNER GLOVES

ACTIVITY

DEMONSTRATE THE PROPER DONNING AND DOFFING OF PPE BASED ON THE TYPE
OF BSL ORGANISM BEING ASKED

RUBRICS

CRITERIA SCORE
DONNING (10)
PROPER ORDER OBSERVED (5)
PROPER APPLICATION OBSERVED
(5)
DOFFING (10)
PROPER ORDER OBSERVED (5)
PROPER APPLICATION OBSERVED
(5)
 CENTRO ESCOLAR UNIVERSITY
SCHOOL OF MEDICAL TECHNOLOGY
MOLECULAR BIOLOGY
LABORATORY

POST LAB QUESTIONS (10pts)

1. Aside from protecting lab workers, what other importance does Biosafety provide?
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2. What is the difference between Biosafety from Biosecurity?

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PREPARED BY: PETER V. SAMONTE, RMT, MSc, MLS(ASCPi)CM

REFERENCES
1. WHO. World Health Organization Biosafety Series. https://www.who.int/ihr/publications...deo-
series/en/
2. https://www.osti.gov/servlets/purl/1658519