HAZARDS IN THE WORKPLACE: BIOHAZARDS

The Clinical Laboratory contains a variety of hazards, many of which are capable of producing serious injury or life-
threatening disease

Definition of Terms:
a. Hazards
- Refers to anything in the environment that has the potential to cause harm
b. Risk
- Possibility of something bad or unpleasant (such as an injury or loss) will happen

For example, a sharp needle is a hazard, but if no one is using it, the needle will not pose any risk.
c. Occupational Safety and Health Act (OSHA)
- Aims to provide all employees (clinical laboratory personnel included) with a safe work environment
- Enacted by the US congress in 1970 but has widely been adopted and used as a basis by other countries to
come up with their own regulations concerning safety in the workplace.

d. Occupational Safety and Health Administration (also OSHA)

- Governing body responsible for ensuring and monitoring the implementation of the standards set by the above-
mentioned act
o Conduct on-site inspections
o Determine whether an employer is complying with the mandatory standards

BIOHAZARDS

Biohazards
- Include all pathogen/ disease-causing microorganisms and the toxins that they produce

Biorisk
- Risk associated to biological toxins or infectious agents

Understanding how microorganisms are transmitted (chain of infection) is essential to preventing infection

CHAIN OF INFECTION
A. 6-part model

Components:
i. Infectious agent / Pathogen
ii. Reservoir
o Animate/ inanimate object where the infectious agent is found normally living
o E.g: Human reservoir, Animal reservoir, Inanimate reservoir
iii. Portal of exit
o How the microorganism escaped from the reservoir

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 iv. Mode of Transmission
o Method of transmission/ route of transmission
o Types
a. Contact Transmission
i. Direct contact
 Actual contact/ close proximity of the infected individual and the susceptible host
 Types:
o Horizontal Transmission: Transmission from a person to person within a group;
No specificity when it comes to the involved hosts
o Vertical Transmission:
 “Mother-to-baby” transmission
 Types:
 Prenatal/ Transplacental Transmission: Transfer of infectious
agent happens BEFORE birth usually by crossing the placenta
 Perinatal Transmission: Transfer happens during passage
through the birth canal

ii. Indirect contact

 Types:
o Transmission by Fomites
 Fomites: Non-living object that may transmit an infectious disease (E.g.
tissues, handkerchiefs, towels, doorknobs, bedding, etc. )
o Droplet transmission
 Droplets: Respiratory particles of moisture containing an infectious agent
 Usually droplets having diameter of >5 micrometers and capable only of
traveling short distances (<1 meter)

b. Common Vehicle Transmission

 Transfer of infectious agents via a medium (soil, water, air)
 Types:
 Airborne transmission
o Pathogen is spread through droplet nuclei (remains after evaporation of
droplets)
o Droplet nuclei have diameter less than 5 micrometers and capable of travelling
distances greater than 1 meter
NOTE DIFFERENCE BETWEEN DROPLET AND AIRBORNE TRANSMISSION
 Waterborne transmission
 Soil-borne transmission
c. Vector borne Transmission
 Vector: Invertebrates capable of harbouring infectious agent
 Types of vectors:
 Mechanical vector: Carry and transport infectious agent; Agent WILL NOT develop while
being transported by the vector
 Biological vector: Infectious agent WILL develop while being transported by the vector

v. Portal of entry
 Routes through which the pathogen enter the host
 Preferred portal of entry: If not followed, will result to milder form of a disease/ non-development
of disease
 E.g. Development of cholera after the causative agent enters via the gastrointestinal
tract; Development of pulmonary tuberculosis after the causative agent enters via the
respiratory tract
vi. Susceptible host

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 B. 3-part model

BIOSAFETY
Biosafety vs. Biosecurity
Biosafety protects people from germs while biosecurity protects germs from people.
Biosafety
o WHO: “Containment principles, technologies, and practices that are implemented to prevent unintentional
exposure to pathogens or toxins, or their accidental release”
Biosecurity
o WHO: “The protection, control, and accountability for valuable biological materials within the laboratories,
in order to prevent their unauthorized access, loss, theft , misuse, diversion, or intentional release

Guidelines & regulations set by the Centers for Disease Control and Prevention (CDC) to prevent exposure to Biohazards
A. Universal Precautions (UP)
o Treats all blood and blood-contaminated samples are potentially infectious
o Main problem: Did not treat other bodily fluids that are not visibly contaminated with blood as potentially
infectious

B. Body Substance Isolation (BSI)

o Consider all body fluids and moist body substances to be potentially infectious
o Main problem: Do not recommend handwashing following removal of gloves unless visual contamination
is present

C. Standard Precautions (SP)

o Combined major features of UP and BSI
o Essentials include proper handwashing, using personal protective equipment (PPE), and proper
environmental control

Essentials of Standard Precautions

A. Handwashing
o Single most effective way of controlling the spread of infectious diseases
o WHO: Scrubbing of the hands should last for at least 20 seconds
o DOH: Should last for at least 15 seconds (2 Happy Birthday Songs)
o Steps:
 Wet your hands with clean, running water. Turn off the tap and apply soap.
 Lather your hands by rubbing them together with the soap.
 Scrub your hands for at least 20 seconds

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 *Proper Scrubbing of Hands

1. Rub hands palm to palm 2. Right palm over left dorsum with 3. Palm to palm with fingers
interlaced fingers and vice versa interlaced

4. Backs of fingers to opposing 5. Rotational rubbing of left thumb 6. Rotational rubbing, backwards and
palms with fingers interlaced clasped in right palm and vice versa forward with clasped fingers of right
hand in left palm and vice versa
 Rinse hand with water
 Dry hand with single-use towel and use towel to turn off the faucet

B. Personal protective equipment (PPE)

o Protective clothing, helmets and other garments designed to protect the wearer’s body from injury or
infection
o Include:
 Laboratory gown, Scrub suits, Respirators, Medical mask, Goggles, Face shield, Head cover,
Rubber boots and overshoes
WHO GUIDELINES: Steps to Put On PPE
1. Put on the scrub suit in the changing room
2. Put on rubber boots. If not available, make sure you have closed, puncture and fluid resistant shoes and
put on overshoes
3. Place the impermeable gown over the scrubs
4. Put on face protection
a. Put on a medical mask
b. Put on goggles or a face shield
5. If available, put a head cover
6. Perform hand hand washing
7. Put on gloves (over cuff)
8. If an impermeable gown is not available, place waterproof apron over gown
WHO GUIDELINES: Steps to remove PPE
1. Remove waterproof apron and dispose of safely. If the apron is to be reused, place it in a container with
disinfectant.
2. If wearing overshoes, remove them with your gloves still on (If wearing rubber boots, see step 4)
3. Remove gown and gloves and roll inside-out and dispose of safely
4. If wearing rubber boots, remove them (ideally using the boot remover) without touching them with your
hands. Place them in a container with disinfectant.

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 5. Perform hand hygiene
6. If wearing a head cover, remove it now (from behind the head)
7. Remove face protection
a. Remove face shield or goggles (from behind the head)
b. Remove mask from behind the head. When removing the mask, untie the bottom string first and
top string next
8. Perform hand hygiene

C. Biosafety Cabinets (BSC)

o Device that encloses a workplace in such a way that protects the workers from exposure to aerosols that
may potentially contain infectious disease agents
o Air that may contain infectious agent is sterilized by passing through the HEPA filter
*HEPA filter
 High Efficiency Particulate Air Filter
 Pores having a diameter of 0.3 um
 Removes air-suspended materials having diameter greater than 0.3 um; Capable of removing up
to 99.97% of air-suspended materials
o Classes:
a. Biosafety Cabinet Class I
 Aka Open Front BSC
 Principle:
o Allow room (unsterilized) air to pass into the cabinet and around the working area
and the materials within
 Does not protect the material and sample from possible contamination
o Before being exhausted from the cabinet: Air passes through HEPA filter,
sterilizing the air to be exhausted
 Protects the worker and the environment from potentially infectious
aerosols

b. Biosafety Cabinet Class II

 Aka vertical laminar flow hoods (technically a misnomer since it is flow hoods are
different from BSCs
 Principle:
o Air passes through HEPA filter BEFORE flowing over the materials within
 Protects the specimen from contamination
o Air to be exhausted first passes through HEPA filter
 Protects the worker and the environment from potentially infectious
aerosols

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 Difference between Biosafety Cabinet Class I and Biosafety Cabinet Class II
Biosafety Cabinet Protection of sample from Protection of the worker from infectious
contamination aerosols
I No Yes
II Yes Yes

 2 Types of BSC II
o Class IIA: 70% of air is recirculated to the working area
o Class IIB: Majority of the air is exhausted and is NOT recirculated

c. Biosafety Cabinet Class III

 Completely enclosed and are equipped with glove ports; Infectious material within is
handled with rubber gloves that are attached and sealed to the cabinet

BIOSAFETY LEVELS (Based on CDC Biosafety in Microbiological and Biomedical Laboratories)

1. Biosafety Level I
o Minimal Risk; Includes the essentials of Standard Precautions
o Does not utilize Biological Safety Cabinets, only needs hand washing facilities
o Biosafety Level 1 agents:
 Include those that have no known potential for infecting people
 Used in laboratory teaching exercises for beginning-level students of microbiology
 Example: Bacillus subtilis
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 2. Biosafety Level II
o Moderate Risk; Includes all those in BSL 1 practices
o In addition to BSL 1 practices, includes the following principles:
 Limiting access to the laboratory during working procedures
 Training laboratory personnel in handling pathogenic agents
 Direction by competent supervisors
 Performing aerosol-generating procedures in BSC Class I or BSC Class II
o Biosafety Level 2 agents:
 Common human pathogens ; Includes those that are being commonly sought in clinical
specimens
 Examples; Hepatitis B ( BSL II requires Hepa B vaccination among all workers), HIV, Salmonella

3. Biosafety Level III

o High risk; Includes all practices listed in BSL 2
o In addition to BSL 2 practices, includes:
 Controlled access
 Decontamination of all wastes
 Decontamination of laboratory clothing before laundering
 BSC Class I, Class II, or Class III
o Biosafety Level 3 agents:
 Indigenous or exotic agents that may cause serious or lethal disease primarily via inhalation of
aerosols
 Includes:
 Viruses that are unlikely to be met in a routine clinical laboratory
 Cultures growing Mycobacterium tuberculosis
 Mold stages of systemic fungi

4. Biosafety Level IV
o Extreme risk; Includes the practices listed in BSL 3
o In addition to BSL 3 practices, also includes:
 Clothing change before entering
 Shower on exit
 Special protective clothing, BSC Class III
o Biosafety Level 4 agents:
 Pose life- threatening risks, maybe transmitted via aerosols
 Infections are frequently fatal, for which there are no vaccines or treatments
 Include only certain viruses of the arbovirus, arenavirus, filovirus group

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 BSL Agents
1 Not known to consistently cause
diseases in healthy adults
2  Agents associated with
human disease
 Routes of transmission
include percutaneous injury,
ingestion, mucous membrane
exposure
3 Indigenous or exotic agents that
may cause serious or potentially
lethal disease through the
inhalation route of exposure
4  Dangerous/ exotic agents
which post high individual risk
of aerosol-transmitted
laboratory infections that are
frequently fatal, for which
there are no vaccines or
treatments
 Agents with a close or
identical antigenic
relationship to an agent
requiring BSL-4 until data are
available to redesignate the
level
 Related agents with unknown
risk of transmission
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Practices
Standard microbiological practices
BSL-1 practices plus:
 Limited access
 Biohazard warning signs
 “Sharps” precautions
 Biosafety manual defining
any needed waste
decontamination or medical
surveillance policies
BSL-2 practice plus:
 Controlled access
 Decontamination of all waste
 Decontamination of
laboratory clothing before
laundering
BSL-3 practices plus:
 Clothing change before
entering
 Shower on exit
 All material decontaminated
on exit from facility
Biohazard and Biosafety
Primary Barriers and Facilities
Safety Equipment
 No primary barriers required Laboratory bench and sink
 PPE: Laboratory coats and gloves; required
eye, face protection, as needed
Primary barriers: BSL-1 plus:
 BSCs or other physical  Autoclave
containment devices used for all
manipulations of agents that
cause splashes or aerosols of
infectious materials
 PPE: Laboratory coats, gloves,
face and eye protection, as
needed
Primary barriers: BSL-2 plus:
 BSCs or other physical  Physical separation from
containment devices used for all access corridors
open manipulations of agents  Self-closing, double-door
 PPE: Protective laboratory access
clothing, gloves, face, eye and  Exhausted air not
respiratory protection, as needed recirculated
 Negative airflow into the
laboratory
 Entry through airlock or
anteroom
 Hand washing sink near
laboratory exit
Primary barriers: BSL-3 plus:
 All procedure conducted in Class  Separate building or
III BSC isolated zone
 If Class III BSC is not available:  Dedicated supply and
Class I or II BSCs in combination exhaust, vacuum, and
with full-body, air-supplied decontamination systems
positive pressure suit
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In addition to Classification by Biosafety Levels, infectious agents are also classified by risk groups.

Risk Group Classification NIH Guidelines WHO Laboratory Biosafety Manual

Risk Group 1 Agents not associated with disease in healthy adult humans
Risk Group 2 Agents associated with human disease that is rarely serious
and for which preventive or therapeutic interventions are
often available.
Risk Group 3 Agents associated with serious or lethal human disease for
which preventive or therapeutic interventions may be
available (high individual risk but low community risk).
Risk Group 4 Agents likely to cause serious or lethal human disease for
which preventive or therapeutic interventions are not usually
available (high individual risk and high community risk).
No or low individual and community risk
A microorganism unlikely to cause human or animal disease.
Moderate individual risk; low community risk
A pathogen that can cause human or animal disease but is
unlikely to be a serious hazard to laboratory workers, the
community, livestock or the environment. Laboratory exposures
may cause serious infection, but effective treatment and
preventive measures are available and the risk of spread of
infection is limited.
High individual risk; low community risk
A pathogen that usually causes serious human or animal
disease but does not ordinarily spread from one infected
individual to another. Effective treatment and preventive
measures are available
High individual and community risk
A pathogen that usually causes serious human or animal
disease and can be readily transmitted from one individual to
another, directly or indirectly. Effective treatment and preventive
measures are not usually available

**Bioterrorism
- Aka Biological attack
- Biological agents (microbes or toxins) used as weapons to further personal or political agendas

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