Principles of Med Lab Science 1

WEEK NUMBER 7 / VIDEO LECTURE (MR. MATT S. ATTAWAY) / TRANSCRIBED BY: NIÑA DE LA CRUZ & ANGEL ULBE
BASIC CONCEPTS OF LABORATORY BIOSAFETY BIOSECURITY
AND BIOSECURITY I “PROTECT BAD BUGS AGAINST BAD PEOPLE"

TOPIC OUTLINE  The aim is to protect these biohazardous or

1 Biosafety vs Biosecurity microbiological agent from the deliberate misuse or
2 Protection of the Laboratory Worker release by bad people. So some people may have
3 Biosafety Cabinets sinister thoughts/plans in sabotaging governments
4 Levels of Biosafety or institutions that they may get access to diseases
5 Spill Management and try to release them to the community or to the
public.

BIOSAFETY VS BIOSECURITY  Small pox is a very dangerous and deadly disease

before, but it has been eradicated through
BIOSAFETY vaccination efforts (small pox doesn’t exist
 Application of safety precautions that reduce a anymore). However, there are still strains or samples
laboratorian's risk of exposure to a potentially of small pox that are kept in high biosecurity
infectious microbe and limit contamination of the facilities.
work environment and, ultimately, the community.
 For example, what if there are people that are
crazy enough to gain access to these diseases and
BIOSECURITY
spread it to the community.
 The discipline addressing the security of
microbiological agents and toxins and the threats
posed to human and animal health, the
environment, and the economy by DELIBERATE
misuse or release. SVERDLOVSK ANTHRAX LEAK

SOVIET CITY OF SVERDLVOSK

 Like in a James Bond film's antagonist, they stop  Now called Ekaterinburg, and is roughly 850 miles
security of microbiological agents and toxins to east of Moscow.
destabilize countries and threaten government.
Biosecurity should be taken seriously. ANTHRAX LEAK
 On April 2, 1979, there was an unusual ANTHRAX
Fortunately, stringent policies and measures are always OUTBREAK initially attributed to tainted meat.
in place when there are risks of these kinds of breaches.
Therefore, the probability of biosecurity breach from  It affected 96 people and killed at least 64 of them
happening is relatively low than the risk of a breach in in the Soviet City of Sverdlovsk.
biosafety
 The first victim died after four days; the last one
 As future laboratory technicians or laboratory died six weeks later.
technologists, one must learn to uphold these measures
in order to protect yourselves and your community WHO CAUSED THIS?
 Soviet officials - They claimed that the deaths
SIMPLE STATEMENTS TO DIFFERENTIATE THE TWO were caused by "intestinal anthrax from tainted
TERMS meat", a story some influential American Scientists
BIOSAFETY also found believable.
"PROTECT PEOPLE AGAINST BAD BUGS"
 They are really the ones at fault according to the
 Bad bugs are your microbiological or confession of the Soviet president 13 years later
biohazardous agent.
PRESIDENT CARTER OFFICIALS
 Biosafety is only concerned about the protection  They suspected that the "outbreak was caused by
of the laboratory worker from these microbiological an accidental release of Anthrax from a suspected
agents, natural protection of people against the bad biological weapons facility in the city.”
bugs because as future laboratory workers, you will
be manipulating these microbiological agents.  The Carter administration accused the Soviet
government that the anthrax outbreak was not
caused by the tainted meat but by a biological

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Principles of Med Lab Science 1
WEEK NUMBER 7 / VIDEO LECTURE (MR. MATT S. ATTAWAY) / TRANSCRIBED BY: NIÑA DE LA CRUZ & ANGEL ULBE
weapons facility. In the side of the Soviet PRIMARY SECONDARY
government, they could also accuse that the Carter CONTAINMENT CONTAINMENT
administration that they were trying to be political  Provides immediate  Intended to protect
because the Soviet government and America at this protection to workers in the laboratory worker,
time were not very friendly with each other. the biological the community, and the
laboratory from environment from
 It wasn't until 13 years later in 1992, the Soviet exposure to chemical unintended
President, Boris Yeltsin, admitted, without going into and biological hazards. contamination with a
details, that the Anthrax outbreak was a result of biological hazard.
military activity at the facility. Around that time,
Yetsin admiited that the military facility was  Examples would be  Facility architectural
responsible for the incident. BSCs (Biosafety features, Facility
cabinets), Ventilation mechanical system
 Russia allowed a team of Western scientists to go Equipment, Animal
to Sverdlvosk to investigate the outbreak. However, Isolation Cages, Vials,  Physical structure of
when they allowed the scientists to go there, they Flasks the facility will serve as
have already made some cover-ups about the the secondary
incident. containment.

 After completing the investigation, the Western  For example, if your

team of scientists concluded that the outbreak was facility is placed in a
indeed caused by release of an aerosol of Anthrax location far from
pathogen at the military facility, but they were still residential areas. If your
unable to determine what specific activity caused facility is in a a gated
the release. community and it has
follow the strict DOX
system for the
 It goes to show the roles that laboratory personnel management of the
plays in the immediate community where the practices ventilation
or deviation of set standards can lead to grave
repercussions that may affect lives permanently  DOX system =
secondary
 The number of deaths weren't minimal containment

 It makes you think what could have happened if it  Ventilation

was accidental or deliberately done to the community. equipment =
primary
 It is also worth noting based from the historical event containment
that governments could really have a degree of cover-
up just to save face.

 As future laboratory personnel, you just need to be

able to have the best practice so none of these will PROTECTION OF THE LABORATORY WORKER
happen
 As part of the primary containment, the protection
of the laboratory worker is of utmost importance, so
CONTAINMENT LABORATORY DESIGN manipulation of biohazards and microbiological agents
cannot be done without the intervention of the
 It is important to understand that a laboratory has a laboratory worker.
containment design. For this lecture, we would be
focusing heavily on primary containment as it is where  It is important that they are protected when they are
we, laboratory scientists or technologists, are most doing their tasks or duties.
involved with

 Containment – refers to the measures that would

keep these microbiological or biohazardous agents
from getting into the community of environment

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Principles of Med Lab Science 1
WEEK NUMBER 7 / VIDEO LECTURE (MR. MATT S. ATTAWAY) / TRANSCRIBED BY: NIÑA DE LA CRUZ & ANGEL ULBE
LABORATORY COVERALLS
PERSONAL PROTECTIVE EQUIPMENT (PPE)

 Worn to minimize exposure to hazards that cause

serious workplace injuries and illnesses

 Includes eye or face protection, body protection,

gloves, respiratory protection

I. EYE AND FACE PROTECTION

SAFETY GOGGLES

 Advances in the PPE design has led to the modern

coveralls

 Offer more protection than laboratory gowns

 Commonly used in chemistry laboratory and not because they fully cover the body
routinely used in the clinical lab
 It is disposable and is able to cover more areas of
FACE SHIELDS the body

 Advantage of wearing one is that there are no

risks of getting contamination when performing your
tasks and or getting infected by something because
lab gowns are usually reworn

 Most often used face protection because it fully III. GLOVES

covers the face, unlike safety goggles  To be protecting your hands which you will be
using to manipulating microbiological agents, gloves
 It is routinely used in the lab, especially with the are necessary
ongoing COVID-19 pandemic.
 You also need to wear gloves when you prepare
 Medical Technologists are the one that perform stains. Stains are used to dye tissue and blood. As
the nasopharyngeal swab and oropharyngeal swab MTs, we need to dye tissues and blood so that there
for those who would like to be tested for RT-PCR for would be distinction to help us better identify these
COVID-19. The Medical Technologists collecting the structures.
samples will be wearing face shields as well.
NITRILE OR LATEX GLOVES

II. BODY PROTECTION

LABORATORY GOWNS

 Most common type

 Are a form of clean gloves in the clinical labs

 There are also surgical gloves, but they are sterile

 One of the most common body protection and used in surgery only

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Principles of Med Lab Science 1
WEEK NUMBER 7 / VIDEO LECTURE (MR. MATT S. ATTAWAY) / TRANSCRIBED BY: NIÑA DE LA CRUZ & ANGEL ULBE
CHEMICAL RESISTANT GLOVES SURGICAL MASKS

 Offer the least amount of protection but the level

 These are not routinely used in performing of protection offered by it is usual enough to prevent
laboratory tests but it is of great use when preparing respiratory transmission of disease
reagents for testing as most reagents used in the
laboratory tests are very strong chemicals.  You just have to wear it properly

 Even with the advent of technology, we still N95/KN95 MASKS

sometimes need to prepare solutions in the
laboratory

 To preserve biopsy specimen and tissue, you need

to have formalin. In some laboratories, you need to
prepare the formalin concentration that best
preserve these biopsy or tissue specimens. Usually,
in order for the laboratory to save money, the
administration will buy the concentrated formalin
and it will be diluted. Here, you will need to wear  They are routinely used in the laboratory unlike
chemical resistant gloves to prepare. the respirators

 Generally, these gloves are not used in the clinical  Offer more protection than surgical masks
lab but is used in preparing reagents
 They are called N95 because the filter out 95% of
very small particles that are about 0.3 microns in
size.
IV. RESPIRATORY PROTECTION
 It is also a must since the nose and mouth is the  N95 and KN95 have the same filter capability. The
most common route by which infectious agents may difference between the two is N95 are approved to
enter your system be used in the medical settings in the US. While the
KN95 are the Chinese standards. Before, KN95
 The most important thing about masks is wearing weren't allowed to be used in the US. Unfortunately,
it properly. It doesn't matter If you have a lot of due to the declining supply of PPEs, especially with
masks but if you don't wear it properly, it won’t give N95 respirators, the CBC has eventually approved
you the optimum protection that it could offer the use of KN95 masks in the US.

RESPIRATORS  Unlike the surgical masks, N95 and KN95 has to

be fitted

 Usually, it would be bought directly. However, in

the clinical setting (hospitals), there is sizing so that
it would be fitted accurately on clinical or hospital
worker

 Usually used in biosafety level 3 or 4 laboratories

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Principles of Med Lab Science 1
WEEK NUMBER 7 / VIDEO LECTURE (MR. MATT S. ATTAWAY) / TRANSCRIBED BY: NIÑA DE LA CRUZ & ANGEL ULBE
USE OF FILTERED TIPS
EQUIPMENT TO PREVENT EXPOSURE

PIPETTES
 Has a removable tip

 It is used to transfer liquids

 Before, it was acceptable that the method of

transferring liquids was through mouth pipetting.
Here, there would be a glass pipette and it would be
attached to a rubber tubing. To suck up the liquid,
you need to put the rubber tubing in your mouth to
draw up so that the fluid will rise. It was acceptable
before but now, it is prohibited because of the risk of
getting disease.

 We now have the modern pipettes  These are disposable

MICROPIPETTES  When you pipette fluids, it will just stay in the

 It is called as such as it is able to dispense very filtered tips
small amount of volumes because in the clinical labs,
we only need small amounts to perform testing. FINGER REST

 1 mL = 1 thousand microliters is more than enough

to perform tests

FEATURES
LESS RESISTANT PLUNGERS

 For ergonomics, it wouldn't slip in your hand.

Thereby, you wouldn’t drop or spill fluids

TIP EJECTORS

 Plunger is the top part

 You press this part to dispense liquid or release to

suck up the solution or fluid

 It needs to be non-resistant so that you will not

really exert much force and that when we pipette, it
will be precise

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Principles of Med Lab Science 1
WEEK NUMBER 7 / VIDEO LECTURE (MR. MATT S. ATTAWAY) / TRANSCRIBED BY: NIÑA DE LA CRUZ & ANGEL ULBE
 It is incorporated with the blue IMBALANCE SENSOR (not seen in the picture)
 When you are going to centrifuge one test tube,
 You just have to press it and the pipette tip will you need to put a counterbalance on the opposite
automatically be detached from the pipette body so side so it would be balance when it turns.
you can directly discard the tip and you don’t have
to physically touch it. Then you can use another filter  If it detects that there is an imbalance, it will give
tip to pipette another solution a warning and it wouldn't push through the
centrifugation process
 Less contact, less exposure

BIOSAFETY CABINETS
CENTRIFUGE
 Is an essential part in the clinical laboratory
because a lot of test would require the preparation
of samples. To prepare samples, you need a
centrifuge because some tests would require you to
separate blood components, such as serum plasma,
red cells

 To separate those blood components, you need to

centrifuge them so that it would be fast. You can
always opt to let them stand But it would take hours
that it would be separated by means of gravity

FEATURES  The terms biological safety cabinet or biosafety

LOCKING LIDS IF ROTOR IS MOVING cabinet has been widely used to describe a variety of
containment devices equipped with HEPA filter(s),
designed to protect personnel from biohazard

 30% of the class II Biosafety cabinets installed in

Southeast Asia are poorly designed, incorrectly
installed, not verified and maintained, and not operated
properly (Whistler T. et al, Applied Biosafety, JABSA
2016, 21, 121-127)

 These lids will lock when they close and it would  It’s okay if its poorly designed, incorrectly installed,
automatically lock once the rotor will turn not verified and maintained because that’s a problem
of the laboratory management. However, if it is not
AEROSOL PROOF ROTO LIDS operated properly, that’s a problem with the laboratory
worker.

 From the earliest laboratory acquired typhoid

infection to hazards posted by the bioterrorism,
antibiotic-resistant bacteria, and rapidly mutating
viruses threats the workers' safety have stimulated the
development and refinement of workstations on which
infectious microorganisms could be safely handled

 Due to the increasing risk of these infections, it has

 There Is a gasket and when you close the lid, it will been a requisite to find ways to protect the laboratory
perfectly fit like a rubber gasket workers.

 It prevents aerosol from spreading or aerosolizing  Early prototype clean air cubicles were designed to
during centrifugation. But ideally, you need to put a protect the materials being manipulated from
cap on your test tubes. environmental or worker-generated contamination.
Before, they made clean air cubicles to protect

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Principles of Med Lab Science 1
WEEK NUMBER 7 / VIDEO LECTURE (MR. MATT S. ATTAWAY) / TRANSCRIBED BY: NIÑA DE LA CRUZ & ANGEL ULBE
whatever you are trying to manipulate so there won’t  Plenum
be contamination.
 Chamber where air is either purified,
 However, the clean air cubicles weren't designed to released into the duct work or released
protect the worker from the risk associated with the back into the laboratory.
manipulation of potentially hazardous materials.
 The red arrows = inflow
 Meaning to say they have made this clean air
cubicles to protect whatever microbiological  It produces a stream of air that directs the
agent that you are trying to manipulate from microbiological agents or airborne agents
getting contaminated, so the result won’t be on where to go. From there, it will go up to
erroneous. From here, there is the realization the plenum and be filtered where it will be
to protect both things, the microbiological released back into the duct work or into the
agent that we are manipulating from getting laboratory itself.
contaminated and we need to protect the one
manipulating from getting infected.  The filtration system usually is composed of a pre-
filter and a HEPA filter

CLASS I BIOSAFETY CABINET  Green arrow = outside room air

 Can easily come through the front opening

of the cabinet

 A stream of inward air moving into the

cabinet contains aerosols generated during
microbiological manipulation. It will contain
it. It will then pass through a filtration
system that traps all air-borne particles
and contaminants. Finally, clean
decontaminated air is exhausted from the
 Has the most basic and rudimentary design of all cabinet.
biological safety cabinetry available today –
 Naturally, this Class I Air Cabinet allows the entry
 It does not prevent samples being handled in the of outside air into the BSC. There is the possibility of
cabinet from being exposed to contaminants that cross-contamination that may affect experimental
may be present in room air. consistency.

 Equipped with "Inflow" - prevents the aerosol  Consequently, the scope and classification of
generated during microbiological manipulations to Class I Cabinet are limited because of the risk of
escape through the front opening cross-contamination. Now, it is largely considered to
be obsolete so it’s not acceptable. Although, it can be
 Sustainable for working with Microbiological used for educational purposes
agents assigned to biosafety levels 1, 2, and 3
CLASS II BIOSAFETY CABINET

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Principles of Med Lab Science 1
WEEK NUMBER 7 / VIDEO LECTURE (MR. MATT S. ATTAWAY) / TRANSCRIBED BY: NIÑA DE LA CRUZ & ANGEL ULBE
 The Class II Type A biological safety cabinet is the purpose that you are doing, you need to change the
most common Class II cabinet. It is also the most airflow or the manner by which the plenum
common safety cabinet of all the different types recirculates the air back into the system.
available.

 Class I is not usually used because its basic

and the risk of cross-examination is
relatively high. Clinical laboratories tend
not to use them and even if they want to use
them, the policy is that licensing bodies do
not allow you to operate if you are using a
Class I Biosafety so you need to use Class II.
There are also limited or there's not many
laboratories that will have a Biosafety Level
4 that would require a Class III Cabinet, but
Class III are expensive. So that’s why Class
II, which is the middle ground, is the most
common or used.  Green arrow = flow of room air into the biosafety
cabinet
 Has "Inflow"- flows through the front inlet grill
 When it enters the biosafety cabinet, it gets
 Has "Downflow" - vertical laminar inside to the inlet grill and it wont able to
(Unidirectional) HEPA-filtered air stream that contaminate your work area
descends downward, continuously flushes the
cabinet interior of airborne contaminants  Red arrows = inflow

 Suitable for work with microbiological agents  Stream of air that which aims to have the
assigned to BSL 1, 2, and 3. airborne aerosols back into the inlet grill so
that it wont contaminate the biosafety
 It has a common plenum from which 30% cabinet.
of the air is exhausted and 70% is
recirculated to the work area as the  Blue arrow = downflow of air
downflow.
 Usually comprised of the 70% recirculated
 If traced amount of toxic chemicals are air coming from the plenum
employed as an adjunct to microbiological
processes, Type A Cabinets should be  It really cleans or maintain the air curtain to
exhaust ducted. If you are using toxic make sure that there is no circulating
chemicals, you need to change the airborne pathogens while manipulating
recirculation. mircobiological agents. This eliminates the
risk of cross-contamination and risk of
infecting the laboratory worker
 The exhaust should 100% be ducted outside
of the laboratory because its design is to
CLASS III BIOSAFETY CABINET
recirculate 70% of the air as downflow into
the work area. So if you're using toxic
chemicals, then it would recirculate the
toxic chemicals.

 Toxic chemicals are usually not filtered by

HEPA filter because HEPA filter will only filter
microbiological agents.

 Exhaust HEPA Filtration will only removes airborne

aerosols, including biohazards but not chemical
fumes. If you are trying to manipulate
microbiological agents and at the same time, you
are using toxic chemicals for whatever test or

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Principles of Med Lab Science 1
WEEK NUMBER 7 / VIDEO LECTURE (MR. MATT S. ATTAWAY) / TRANSCRIBED BY: NIÑA DE LA CRUZ & ANGEL ULBE
 Provides absolute level of safety, which cannot be Then you close it and perform air
attained with Class I and Class II Cabinets purification or decontaminate it. From
there, you can manipulate the equipment.
 Made of welded metal construction and are
designed to be gas tight.

 Work is performed through glove ports in front

of the cabinet

 Negative pressure relative to the ambient

environment is maintained within the cabinet

 Negative pressure provides a fail-safe

mechanism in case physical containment is
compromised

 A supply of HEPA filtered air provides product  Exhaust air back into the laboratory. However, air
protection and prevents cross contamination of may be exhaust air into a dedicated duct work
samples. Exhaust air is usually HEPA filtered and system into the external environment. They are
incinerated frequently specified for work, involving the most
lethal biological hazards.
 Product is referred to as microbiological
agents  As you can see, the outside air directly enters into
the plenum. It has nowhere to enter through the front
 Sometimes, these scientists are working on opening anymore. The only opening is through the
microbiological agents that can be also glove ports, whichare gas-tight
considered as products. They are trying to
manufacture products, such as vaccines or  There is inflow and downflow
antibiotics. Some antibiotics can be derived
from microbiological agents, which can be  There is a dunk tank
considered as products.

 The level of safety on this cabinet shouldn’t

be taken lightly so there is strong protection WORKING IN A BIOSAFETY CABINET

 Exhaust air is usually HEPA-filtered and  As medical laboratory scientists or technicians, we

incinerated because if there are airborne would be usually working in a BSC but not usually in a
pathogens, it would be killed. If incineration Class III BSC but most often, you will be working in a
is not possible, then the alternative way is Class II BSC.
to do double HEPA filtration in series.

 Suitable for work with microbiological agents

assigned to BSL (Biosafety Level) 1,2,3, and 4.

 Materials are transferred into the cabinet through

a pass-through unit or inducted tank, which is
installed on the side of the work area.

 Since its gas tank, it is closed and can be

accessed through glove ports. The only way
for you to get your materials or
microbiological agents from the outside in
order to bring it inside the safety cabinet is
through a dunk tank or a pass-through unit

 This is where you will put materials or other

equipment, such as wireloops or pipette.

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Principles of Med Lab Science 1
WEEK NUMBER 7 / VIDEO LECTURE (MR. MATT S. ATTAWAY) / TRANSCRIBED BY: NIÑA DE LA CRUZ & ANGEL ULBE
BEFORE USE 2 CLEAN AS YOU GO
1 CLEAN INTERIOR PANELS AND 3 PROTECT THE AIR CURTAIN
MATERIALS/ITEMS PLACED INSIDE  If it’s capable of inflow or downflow, you need
 You need to clean them before you put them to protect the areas wherein air streams are
inside flowing. You need to not block those vents that
are producing these air streams. If you block
2 PURGE AIR FOR 3+ MINUTES them, you are effectively blocking the capacity of
 These biosafety cabinets have this feature the BSC to flush these airborne pathogens, which
where you can purge the air or filter. So you can can cause contamination in whatever you are
remove the air contaminants, that are present in working with
the air
4 DO NOT USE FLAME INSIDE THE BSC
3 ADJUST SASH TO WORKING HEIGHT How do you decontaminate wireloops?
 There is a glass covering to protect you from
the microbiological agent or from getting  There are small incinerators that you can use
infected with whatever you are working with. to put in your metal wireloops so it would be
heated, which leads it to be decontaminated or
 It can be raised up or down. Fully down to sterilized.
close it and purge the air. After purging, you
need to raise it to the working height. The red line  Before, the risk of having an alcohol lamp
indicates the level to which it is designed to be inside the BSC is that you sometimes forget to
raised when you are working with the BSC. turn on the air streams, then it would become
too hot. If the work area becomes too hot, it
 This is the area where you will place your might damage the HEPA filters and could lead to
hands or materials. Before manipulating the an ineffective filtering of the air. You may think
microbiological agents, you must make sure to that the BSC is working properly but in reality, it
place everything you need inside. is not so you render yourself to be infected.

 It happened before in Velez that a BSC got

caught in fire because an intern forgot about an
alcohol lamp that was turned on inside the BSC

AFTER USE
1 CLEAN INTERIOR PANELS AND ITEMS STORED
OUTSIDE
 Before removing them from the BSC, you
need to clean them inside the BSC.

2 PURGE AIR FOR 3+ MINUTES

3 TURN ON UV IF SAFE
 It has a UV lamp or feature to decontaminate
the workstation or the insides of the cabinet

4 ACCOMPLISH USAGE LOG

 So, if you have usage logs, you need to fill them
up so that you are able to document what you
did inside the BSC. If there are damages or
DURING USE changes in performance of the cabinet, you can
1 DESIGNATE CLEAN, WORKING, AND DIRTY take note.
AREAS
 So that there would be a level of organization,
you need to assign these areas. It can be from
left to right and vice-versa depending on which
direction you are comfortable with

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Principles of Med Lab Science 1
WEEK NUMBER 7 / VIDEO LECTURE (MR. MATT S. ATTAWAY) / TRANSCRIBED BY: NIÑA DE LA CRUZ & ANGEL ULBE
HEPA FILTERS  Continuous filter sheet or HEPA filter is split
or folded
 Early versions of ventilated cabinets did not have
adequate or controlled directional air movement. They  Aluminum separator = corrugated
were characterized by mass air flow into the cabinets. aluminum structures are placed to give
Albeit, with widely varying volumes across openings. shape to the filter so it wont stick with one
another
 Mass air flow in the cabinet grow contaminated air
away from the laboratory worker. So this was the  Filter frame = to give its shape. Square or
forerunner of the Class I Biosafety Cabinet. However, rectangular structure
since the air was unfiltered, the cabinet was
contaminated with environmental microorganisms and
 Found in all BSC classes
other undesirable particulate matter.
 Although, there are some biosafety cabinet
 It has come to a point that they wanted to protect
level I that do not have HEPA filters.
whatever microbiological agent that they are working
Generally, modern biosafety cabinets have
with from getting contaminated so that’s why they
these HEPA filters
made the clean air cubicles. Next, they wanted this
protect the laboratory workers as well. They had the
mass air flow produced in the cabinets. However, the
LEVELS OF BIOSAFETY
problem with the mass air flow, coming from the
outside, it contaminated the work, which led to a
dilemma. BIOSAFETY LEVEL 1
 Suitable for work involving agents not known to
 Control of airborne particulate materials became consistently cause disease in immunocompetent
possible with the development of filters. Now, they have adult humans
the idea to filter the air before recirculating it back. This
efficiently removed microscopic materials from the air  Categorizing biosafety level needed based
on what you working with
 HEPA filter was develop to create dust-free working
environments in the 1940s  Present minimal potential hazard to laboratory
personnel and the environment.

 Special containment devices or equipment, such

WHAT IS HEPA FILTER? as BSC, are not generally required

 If you are manipulating agents that needs

to be manipulated in the cabinet, you need
to have it as well

 Maybe used as determined by appropriate

risk assessment. There are no special
practices required here.

 Work is typically conducted on open bench tops

using standard microbiological practices

 It doesn’t require you to have BSC but if

appropriate risk assessment tells you to
 Removes the most penetrating particle size have one, you need to have one.
(MPPS) of 0.3 ϻm (micrometers) with an efficiency of
at least 99.97% BIOSAFETY LEVEL 2
 Suitable for work involving agents that pose
 Disposable dry filter, constructed of borosilicate moderate hazards to personnel and the
microfibers cast into a thin sheet, much like a piece environment
of paper.

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Principles of Med Lab Science 1
WEEK NUMBER 7 / VIDEO LECTURE (MR. MATT S. ATTAWAY) / TRANSCRIBED BY: NIÑA DE LA CRUZ & ANGEL ULBE
 The key indicator in determining the change room and you must change your
biosafety level depends on what type of clothes between the two-self closing doors
agents are being manipulated
BIOSAFETY LEVEL 4
 Access to the laboratory is restricted while work is  Required for work with DANGEROUS and EXOTIC
being conducted agents that pose a high individual risk of aerosol-
transmitted laboratory infections and
 Laboratory personnel have specific training lifethreatening disease that is frequently fatal, for
in handling specific pathogens and are which there are no vaccines or treatments, or related
supervised by scientists that are competent agent with unknown risks of transmission.
in handling infectious agents and
associated procedures  Agents with a close or identical pathogenic
relationship to agents requiring BSL 4
 All procedures in which infectious aerosols or containment must be handled at the BSL 4
splashes may be created are conducted in BSCs or until sufficient data are obtained to either
other physical containment equipment confirm continued work at the BSL 4 level or
redesignate the level of that identical agent
 Laboratory personnel must be provided medical
surveillance, as appropriate, and offered available  Laboratory staff must have specific and thorough
immunization for agents handled or potentially training in handling extremely hazardous infectious
present in the laboratory. agents

 For example, if you are working in BSL 2 and  BSL-4 Cabinet and Suit Laboratories have special
you are handling yellow fever strains, the engineering and design features to prevent
positive agent for yellow fever or hepatitis microorganisms from being disseminated into the
A, you need to be immunized or vaccinated environment.
for those diseases to mitigate the risk of you
getting infected from the manipulation of
those infected agents.

BIOSAFETY LEVEL 3
 Applicable to clinical, diagnostic, teaching,
research, or production facilities where work is
performed with indigenous or exotic agents that may
cause serious or potentially lethal diseases through BIOSAFETY LEVEL 4 LABORATORY MODELS
the inhalation route of exposure SUIT LABORATORY

 Laboratory personnel must have specific training

in handling pathogenic and potentially lethal agents
and must be supervised by scientists who are
competent in handling in infectious agents and
associated procedure

 All procedures involving the manipulation of

infectious materials must be conducted within BSCs
or other physical containment devices.

 A BSL-3 laboratory has special engineering and

design features
 This is where the scientists are wearing a suit and
 One of the special engineering and design provided with air for them to breathe
features require the laboratory to be
separated from areas that are open to  Manipulation of agents must be performed in a
unrestricted traffic flow within the building. class II BSC 2
Laboratory is restricted and the access to
the laboratory is through two self-closing  Personnel must wear a positive pressure
doors. A clothing change is required. You supplied air protective suit.
are provided with an ante room or a

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Principles of Med Lab Science 1
WEEK NUMBER 7 / VIDEO LECTURE (MR. MATT S. ATTAWAY) / TRANSCRIBED BY: NIÑA DE LA CRUZ & ANGEL ULBE
 All manipulations of infectious agents must be
performed within a biosafety cabinet or other
primary barrier system

 Workers must wear laboratory clothing, such as

scrub suits, before entering the room used for
donning positive pressure. All laboratory clothing
must be removed in dirty site change room before
entering the personnel shower

 You cannot wear your outside clothes and

wear the positive pressure suit. You need to
change into another article of clothing, such
as scrubs, then you wear the positive
pressure suit.

 The suit is already prepared with gloves,

but you are still required to wear disposable
gloves underneath to ensure that if there
are tears or breakages in the outer suit, you
can still be protected. Disposable gloves
must not be worn outside the change area.

CABINET LABORATORY BIOSAFETY LEVEL 1

 Skin bacteria are normal bacteria in your skin.
You can study or manipulate them in a BSL-1
Laboratory

 Yeast are also not really clinically significant in

immunocompetent adult humans because as
already known, fungi are not very effective in
infecting humans but only to those who are
immunocompromised.
 Manipulation of agents must be performed in a
class III BSC BIOSAFETY LEVEL 4
 Smallpox already has a vaccine. If you get
 BSL 4 laboratory consists of either a separate infected, it will still pose a life-threatening disease
building or a clearly demarcated and isolated zone
within a building

 Rooms in the facility must be arranged to ensure

SPILL MANAGEMENT
sequential passage through an inner dirty changing
room, a personal shower, and outer clean changing
SPILLS
room upon exiting the rooms.
 One of the most common cause of biohazard
spread is through biological spills.
 It must have a pass-through dunk tank,
fumigation chamber or equivalent decontamination
method so that materials and equipment that SPILL MANAGEMENT
cannot be decontaminated in the autoclave can be  In order to manage any biological spills, you need
safely removed from the cabinets certain materials to effectively or efficiently clean up
any spills
 The dunk tank must be capable of being
fumigated with chemical disinfectants so
that the materials can be clean prior to
entering the BSC Level 3 and prior to SPILL KIT
removing them from BSC Level 3
 An important feature of the laboratory and must
always be updated and available at all times.

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Principles of Med Lab Science 1
WEEK NUMBER 7 / VIDEO LECTURE (MR. MATT S. ATTAWAY) / TRANSCRIBED BY: NIÑA DE LA CRUZ & ANGEL ULBE
 Always updated = it should be regularly  Example: A chemical has spilled, and
replenished. An example would be what if you need to leave the room in order to
there was a spill yesterday and they did not disperse it before you clean up the spill.
replenish the items in the spill kit. If a spill were If you spill formalin, its painful to the
to happen today, the tendency is that you eyes so you need to disperse it for a
cannot efficiently clean the spill. The nature of while.
the agent of that spill should be cleaned up
immediately If liquid spill has contaminated clothing, remove
2 contaminated clothing, turning exposed areas
inward, and place in a biohazard bag.
COMPOSITION OF BASIC SPILL KIT
Concentrated household bleach 3 Wash all exposed skin with soap and water.

 Also known as sodium hypochlorite

1
 Brand names: Zonrox, Chlorox, or CLEAN-UP OF BIOSAFETY LEVEL 2 SPILL
Winrox Allows aerosols to disperse for at least 15
1
minutes before re-entering the laboratory
Spray bottle for making 10% bleach solutions 2 Assemble clean-up materials from spill kit
3 Put on protective clothing
 You need to have the concentrated
household bleach and prepare the 10% Depending on the nature of the spill, it may be
4
solution as you use it because sodium advisable to wear an N-95 Respirator.
hypochlorite or 10% bleach solution is
unstable. So when they are open to air, Pick up any sharp objects with forceps or tongs
5
2 chlorine evaporates at a higher rate and discard in a sharps container
from the solution, rapidly reducing the Cover the area of the spill with paper towels or
concentration of chlorine 6
other absorbent materials

 Chlorine is the one capable killing the Using mechanical means, scoop the absorbed
microorganisms or microbiological 7 spill material and discard in a biohazard bag for
agents. So you need to prepare it fresh subsequent autoclaving

Forceps, autoclave broom, and dust pan, or

other mechanical devices for handling sharps

3  So that you won’t remove the sharp

materials with your hands because you
might harm yourself

Paper towels or other suitable absorbent

4 material
Biohazard bags for the collection of
5 contaminated spill clean-up items

6 Utility gloves and medical examination gloves

7 Face protection
8 Sharps container

WHAT TO DO IN A BIOSAFETY LEVEL (BSL-2)

SPILL
1 If an agent poses an inhalation risk, quickly leave
the room. Notify others to leave. Most agents
used at BSL-2 Level are not airborne pathogens.
Close door, and post with a warning sign.

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