Evolve.

Adapt.
Overcome.
CEFI is now ready.

SAFETY IN THE
MICROBIOLOGY
LABORATORY
PREPARED BY: MA. JOY CHRISTINE R. AGUILAR, RMT, MLS (ASCPi)CM
STANDARD PRECAUTIONS
•The precautions required that all patients and
body fluids except sweat are treated as
potentially infectious and hazardous.
 EXPOSURE CONTROL PLAN
• The plan that identifies tasks that are hazardous to employees and
promotes employee safety. This is the responsibility of the
Microbiology supervisor and Laboratory Director.

• 5 most common laboratory acquired infections:

1. Shigellosis
2. Salmonellosis
3. Tuberculosis
4. Brucellosis
5. Hepatitis
EXPOSURE CONTROL PLAN
• Males and younger employees (17 to 24 years old) are involved
in more laboratory-acquired infections than females and older
employees (45 to 65 yrs. old)
• Post exposure investigation is done by the Microbiology
Supervisor and Safety officer.
• Most important practice to prevent the spread of disease:
Handwashing
The essentials of Standard Precautions and safe laboratory work
practices are as follows:
• Do not eat, drink, smoke, or apply cosmetics.
• Do not insert or remove contact lenses.
• Do not bite nails or chew on pens.
• Do not mouth-pipette.
• Limit access to the laboratory to trained personnel only.
• Assume all patients are infectious for all bloodborne pathogens.
• Use appropriate barrier precautions to prevent skin and mucous membrane exposure, including
wearing gloves at all times and masks, goggles, gowns, or aprons if splash or droplet formation is a
risk.
• Thoroughly wash hands and other skin surfaces after removing gloves and immediately after any
contamination.
• Take special care to prevent injuries with sharp objects, such as needles and scalpels.
ROUTES OF INFECTION
 SAFETY PROGRAM I N THE CLINICAL
LABORATORY
EXPOSURE CONTROL PLAN
-Identifies tasks that are hazardous to employees and promotes employee safety
through the use of the following:
• Employee education and orientation
• Appropriate disposal of hazardous waste
• Standard Precautions
• Engineering controls and safe work practices, as well as appropriate waste disposal and use
of BSCs
• Personal protective equipment (PPE)
• Post-exposure plan involving the investigation of all accidents and a plan to prevent
recurrences.
 EMPLOYEE EDUCATION AND
ORIENTATION
• Each institution should have a safety manual that is reviewed
by all employees and a safety officer who is knowledgeable
about the risks associated with laboratory acquired
infections.
• Hand washing should be emphasized for all laboratory
personnel.
• All employees should also be offered, at no charge, the
HBV vaccine and annual skin tests for tuberculosis.
 DISPOSAL OF HAZARDOUS WASTE
• All materials contaminated with potentially infectious agents must
be decontaminated before disposal.
• Infectious waste may be decontaminated by the use of an
autoclave, incinerator, or any one of several alternative waste-
treatment methods.
• Infectious waste (agar plates, tubes, reagent bottles) should be
placed into two leak-proof, plastic bags for sturdiness; this is
known as double-bagging in common laboratory jargon.
• Pipettes, swabs, and other glass objects should be placed into rigid
cardboard containers before disposal
 DISPOSAL OF HAZARDOUS WASTE
• Broken glass is placed in thick boxes lined with plastic
biohazard bags; when full, the box is incinerated or
autoclaved.
• Sharp objects are placed in Sharps containers, which are
autoclaved or incinerated when full.
 ENGINEERING CONTROLS
- Controls that isolate or remove the hazard from the workplace.

- Examples of Engineering Controls:

• Use of closed tube sampling by laboratory equipment
• Use of safety needles and single-use holders
• Eyewash stations
• Emergency showers
• Plastic shield barriers
 WORK PRACTICE CONTROLS
- Defined by OSHA as “altering how a task is performed to reduce the
likelihood of exposure to infectious agents”.

- Examples of Work Practice Controls:

• No mouth-pipetting
• No eating, drinking, smoking, or applying cosmetics in the laboratory
• Disinfection of workstations at the end of each shift and after any spill of
infectious material
• No recapping or breaking of contaminated needles
• Disposal of needles in an appropriate puncture-resistant container
PERSONAL PROTECTIVE EQUIPMENT (PPE)
- Specialized clothing or equipment that is worn by an employee for protection.
Examples of PPE:
• Plastic shield or goggles • Impervious gowns
• Disposal containers for sharp objects • Laboratory coats
• Holders for glass bottles • Disposable gloves
• Trays in which to carry smaller • Masks
hazardous items (e.g., blood culture • Safety carriers for centrifuges
bottles) (especially those used in the AFB
• Handheld pipetting devices laboratory)
• HEPA respirators
 POSTEXPOSURE CONTROL
• All laboratory accidents and potential exposures must be reported to the
microbiology supervisor and safety officer, who will immediately arrange to
send the individual to employee health or an outside occupational health
physician.
• The safety committee or, at minimum, the laboratory director and safety officer
should review the accident to determine whether it could have been prevented
and to delineate measures to be taken to prevent future accidents.
• Immediate medical care is of foremost importance; investigation of the
accident should take place only after the employee has received appropriate
care.
• The investigation of the accident and corrective action should be documented in
writing in an incident report
 BIOLOGICAL SAFETY CABI NET

• A device that encloses a workspace in such a way as to protect

workers from aerosol exposure to infectious disease agents. Air
that contains the infectious material is sterilized, either by heat,
ultraviolet light, or most commonly, by passage through a HEPA
filter that removes most particles larger than 0.3 µm in diameter.
• BSCs are designated by class according to the degree of biological
containment they afford.
 CLASSES OF Biosafety Cabinet
1. Class I BSC
- Allows room (unsterilized) air to pass into the cabinet and around the area
and material within, sterilizing only the air to be exhausted.
- Has negative pressure, and is ventilated to the outside, and is usually
operated with an open front.
2. Class II BSC
- Sterilize air that flows over the infectious material, as well as air to be
exhausted.
- Also known as vertical laminar flow BSC.
 CLASSES OF Biosafety Cabinet
a. Class IIA
- Self-contained cabinet
- 70% of the air is recirculated.
- Has a fixed opening.
- Most common BSC used in hospital clinical microbiology laboratory.
b. Class IIB
-The exhaust air is discharged outside the building.
- Have a variable sash opening.
- It is selected if radioisotopes, toxic chemicals, or carcinogens will be used
 CLASSES OF Biosafety Cabinet
3. Class III BSC
- Completely enclosed cabinets.
- With negative pressure.
- Afford the most protection to the worker.
- Air coming into and going out of the cabinet is filter sterilized, and the
infectious material within is handled with rubber gloves that are attached and
sealed to the cabinet.
 BIOSAFETY LEVELS
1. BIOSAFETY LEVEL 1 AGENTS
- Agents that have no known potential for infecting healthy people. Used in
laboratory teaching exercises for beginning-level students in microbiology.
- Examples: Bacillus subtilis, Mycobacterium gordonae; Laboratory work
using these agents can be conducted on open benchtops.
2. BIOSAFETY LEVEL 2 AGENTS
- Includes the most common microorganisms associated with laboratory-
acquired infections.
- Examples: Bacillus anthracis, Yersinia pestis, Salmonella, Shigella,
Staphylococcus, HBV, HIV; The recommended BSC is Class II.
 BIOSAFETY LEVELS
3. BIOSAFETY LEVEL 3 AGENTS
- Agents that are either indigenous or exotic. Have the potential for aerosol
transmission.
- Examples: Mycobacterium tuberculosis, Brucella spp., Francisella tularensis,
systemic fungi, rickettsiae, and specific viruses; Has an available vaccine or
therapy.
4. BIOSAFETY LEVEL 4 AGENTS
- Agents that are dangerous and exotic. Have a high risk of causing life-threatening
infections, can be transmitted by aerosols, or have an unknown risk of
transmission. Primarily used in research facilities.
- Examples: Arenavirus, filovirus; The recommended BSC is Class III. Do not
have an available vaccine or therapy.
 CHEMICAL SAFETY
• Hazard Communication Standard (Right to Know Law)
– The lab, by the law must provide written policies for the safe use of hazardous
chemicals (OSHA)
• Material Safety Data Sheet
– Contains information of the nature of the chemical, precautions in handling a
spilled chemical, and disposal recommendations.
• Chemical Hygiene Plan –
• Each laboratory should have a chemical hygiene plan that includes guidelines
on proper labeling of chemical containers, manufacturer’s material safety data
sheets, and the written chemical safety training and retraining programs.
FIRE SAFETY
• Each laboratory is required to post fire evacuation plans that
are essentially blueprints for finding the nearest exit in case
of fire. Exit ways should always remain clear of obstructions,
and employees should be trained to use fire extinguishers.
 FIRE SAFETY
• TYPES OF FIRE EXTINGUISHERS
1. Type A – used for trash, wood, and paper
2. Type B – used for chemical fires
3. Type C – used for electrical fires
4. Type ABC – found in most laboratories
• IMPORTANT ACTIONS IN CASE OF FIRE (RACE)
1. Rescue any injured individuals.
2. Activate the fire alarm.
3. Contain (smother) the fire, if feasible (close fire doors).
4. Extinguish the fire, if possible.
 REGULATORY AGENCIES
• Occupational Safety and Health Administration (OSHA)
• Centers for Disease Control and Prevention (CDC)
• The Joint Commission
• Clinical Laboratory Standards Institute (CLSI)
• College of American Pathologists (CAP)
 CONTROL OF MICROORGANISM
• Sterilization is a process that kills all forms of microbial
life, including bacterial spores.

• Disinfection is a process that destroys pathogenic

organisms, but not necessarily all microorganisms or spores.

Sterilization and disinfection may be accomplished by

physical or chemical methods.
PHYSICAL METHODS OF STERILIZATION

1. Incineration
- Most common method of treating infectious waste
- Hazardous material is burned to ashes at temperatures of
870° to 980°c.
 PHYSICAL METHODS OF STERILIZATION
2. Moist Heat
- Steam under pressure.
- Used to sterilize biohazardous trash and heat-stable objects with the use of an
autoclave.
- Fastest and simplest physical method of sterilization.
- Moist heat in the form of saturated steam under 1 atmosphere (15 psi [pounds per
square inch]) of pressure causes the irreversible denaturation of enzymes and
structural proteins.
- Two most common sterilization temperatures are 121°C (250°F) and 132°C (270°F).
- The autoclave must be monitored periodically to measure its effectiveness.
- Bacillus stearothermophilus (Geobacillus stearothermophilus) spore indicator strips,
which are quite resistant to the effects of the autoclave, can be used for this purpose.
 PHYSICAL METHODS OF STERILIZATION
3. Dry Heat
- Used to sterilize items such as glassware, oil, petrolatum, or powders with the
use of an oven
- Requires longer exposure times (1.5 to 3 hours) and higher temperatures than
moist heat (160° to 180°C)

4. Ionizing Radiation
- Used for sterilizing disposables such as plastic syringes, catheters, or gloves
before use
- Short wavelength and high-energy gamma rays are used in ionizing radiation
from microwaves and radiograph machines.
 PHYSICAL METHODS OF STERILIZATION
5. Filtration
- Method of choice for antibiotic solutions, toxic chemicals, radioisotopes,
vaccines, and carbohydrates, which are all heat sensitive.
- Filtration of liquids is accomplished by pulling the solution through a cellulose
acetate or cellulose nitrate membrane with a vacuum.
- Filtration of air is accomplished using high efficiency parti-culate air (HEPA)
filters designed to remove organisms larger than 0.3 µm from isolation rooms,
operating rooms, and biological safety cabinets (BSCs).
 CHEMICAL METHODS OF STERILIZATI ON
1. Ethylene Dioxide Gas
- Most common chemical sterilant.
- Used to sterilize heat-sensitive objects.
2. Formaldehyde Vapor & Vapor-Phase Hydrogen Peroxide
- Have been used to sterilize HEPA filters in BSCs.
3. Glutaraldehyde
-Used for sterilizing medical equipment
- Sporicidal (kills spores) in 3-10 hours.
- Does not corrode lenses, metal, or rubber.
 CHEMICAL METHODS OF STERILIZATI ON
4. Peracetic Acid
- Has been used for the surface sterilization of surgical instruments.
- Effective in the presence of organic material.

*Cold sterilization refers to the use of glutaraldehyde or peracetic acid

in the sterilization process.
 PHYSICAL METHODS OF DISINFECTION
1. Boiling
- Kills vegetative bacteria
- 100°C for 15 minutes
2. Pasteurization
- Kills food pathogens
- 63°C, 30 min (Batch method)
- 72°C, 15 secs (Flash method)
3. Nonionizing Radiation
- UV rays are long wavelengths and low energy.
- They do not penetrate well and organisms must have a direct surface exposure, such as the
working surface of a BSC, for this form of disinfection to work.
 CHEMICAL METHODS OF DISINFECTION
1. Alcohols
- Ethyl alcohol and isopropyl alcohol : Either is best used on the skin as an
antiseptic or on thermometers and injection vial rubber septa as a
disinfectant.
- Not sporicidal; have poor activity against certain nonenveloped viruses. It
is inactivated by the presence of organic material.
 CHEMICAL METHODS OF DISINFECTION
2. Aldehydes
a. Formaldehyde (Formalin)
- Used to disinfect biosafety hoods. (This type of procedure should be left to professionals.)
- Although it can be used as a chemosterilizer in high concentrations, its usefulness is limited by
its irritability factor and its potential carcinogenicity. Not recommended as disinfectant or
sterilant on a routine basis.
b. Glutaraldehyde
- When used as a 2% solution, it is germicidal in approximately 10 minutes and sporicidal in 3-10
hours.
- Kills microorganisms by inactivating DNA and RNA through alkylation of sulfhydryl and amino
groups.
- Sterilizer of choice for medical equipment that is not heat-stable and cannot be autoclaved
as well as for material that cannot be sterilized with gas.
 CHEMICAL METHODS OF DISINFECTION
3. Halogens
- Frequently used disinfectants.
a. Iodophor
- Combination of iodine and a neutral polymer carrier that increases the agent’s solubility, which
allows the slow release of iodine.
- Povidone-iodine (Betadine) – best-known iodophor; mainly used as an antiseptic
- Commonly used as skin preparations from sites where blood is to be drawn.
4. Heavy Metals
- Rarely used in clinical applications ; Slowly bactericidal; primarily bacteriostatic
- Heavy metals containing mercury are no longer recommended.
- An eyedrop solution containing 1% silver nitrate is still instilled in the eyes of newborns to
prevent infections with Neisseria gonorrhoeae.
 CHEMICAL METHODS OF DISINFECTION
5. Quaternary Ammonium Compounds
- Cationic, surface-active agents, or surfactants, that work by reducing the
surface tension of molecules in a liquid.
- Inactivated by excess organic matter. Limited to disinfection of noncritical
surfaces (e.g., benchtops, floors).
6. Phenolics
- Stable, biodegradable, and relatively active in the presence of organic material.
- At lower concentrations, phenolics are able to disrupt enzyme systems.
- Mainly used in the disinfection of hospital, institutional and household
environments
 THANK YOU!!!
GOOD LUCK ON YOUR EXAMS ☺