Safety and

Specimen
Management
Chapter 4: Laboratory Safety

Gerald John A. Paz, RMT, MLS(ASCPi)

Sterilization and Disinfection

 Sterilization
 process that kills all forms of microbial life, including
bacterial spores.
 Destruction of all forms of life, including bacterial spores.
 all-or-nothing process
Sterilization and Disinfection

 Disinfection
 destroys pathogenic organisms, but not necessarily all
microorganisms or spores.
 eliminates a defined scope of microorganisms, including
some spores
 Disinfectants : for inanimate objects
 Antiseptic : applied to skin for eliminating or reducing
the number of bacteria present
Physical Methods of
Sterilization
 Incineration
 Moist heat
 Dry heat
 Filtration
 Ionizing (gamma) radiation
Incineration

 most common method of treating infectious waste

 Burned into ashes
 Temperature: 870° to 980°C.
 incineration is recommended for elimination of prions
and infective proteins.
 Disadvantages:
 Toxic air emissions
 presence of heavy metals in ash
Moist Heat

 Steam under pressure

 For biohazardous trash and heat-stable objects

 Fastest and simplest physical method of sterilization

Moist Heat

 Autoclave:
 media, liquids, and instruments:
 1 atm of pressure, or 15 psi,
 temperature of 121° C. At this temperature, all
microorganisms (an exception being prions) and their
endospores are destroyed within approximately
 15 minutes of exposure.
 Infectious medical waste:
 132°C for 30 to 60 minutes to allow penetration of the
steam throughout the waste and the displacement of air
trapped inside the autoclave bag.
 important factor is that the moisturized heat comes in
contact with the material
 Additional:
 Autoclave bag:
 Autoclave tape:
 Indicator: Bacillus stearothermophilus
 thermophilic species which can grow at 65°C and above. The
spores are highly resistant and are used to monitor autoclave
performance.
Dry Heat

 Usually used for glassware, oil, petrolatum, or powders

 requires much longer exposure times and higher

temperatures than moist heat
 1.5 to 3 hours
 160°to 180°C
Filtration

 accomplished through the use of thin membrane filters

composed of plastic polymers or cellulose esters
containing pores of a certain size
 For liquids:
 Cellulose acetate or cellulose nitrate membrane with a
vacuum
 liquid is pulled (vacuum) or pushed (pressure) through the
filter matrix

 For air:
 HEPA filters
 remove organisms larger than 0.3 μm from isolation rooms,
operating rooms, and biologic safety cabinets (BSCs)
Filtration

 Most bacteria, yeasts, and moulds are retained by pore

sizes of 0.45 and 0.80 μm; however, this pore size may
allow passage of Pseudomonas-like organisms, and
therefore a 0.22-μm size is available for critical
sterilizing

 Common applications: parenteral solutions, vaccines,

and antibiotic solutions
Ionizing/ non ionizing
radiation
 Ionizing radiation-- short wavelength and high energy;
gamma rays or electron beams

 Non-ionizing radiation– long wavelength and low

energy; ultraviolet light
Ionizing Radiation

 used by the medical industry for the sterilization of

disposable supplies, such as syringes, catheters, and
gloves
Non Ionizing Radiation

 Because of its poor penetrability, the usefulness of

nonionizing radiation is limited; it can be used to
disinfect surfaces, although the parameters (distance to
surface, potential microorganisms to be destroyed)
under which it is to be used need to be determined.
Chemical Methods of
Sterilization
 Ethylene oxide
 Formaldehyde vapor
 Vapor-phase hydrogen peroxide
 Glutaraldehyde
 Peracetic acid
Chemical agents exert their
killing effect by the following
mechanisms:
 Reaction with components of the cytoplasmic
membrane
 Denaturation of cellular proteins
 Reaction with the thiol (–SH) groups of enzymes
 Damage of RNA and DNA
Physical Methods of
Disinfection
 Boiling
 Pasteurization
 Nonionizing radiation
 Add’l
Low temperature holding (LTH)/ batch method:
- doesn’t affect the taste of food
- milk borne pathogens
High temperature short time (HTST)/ Flash Pasteurization
- quick heating and then immediate cooling
Ultra high temperature (UHT)
- 140o C for 3 seconds (cooled very quickly in a
vacuum chamber)
- milk can be stored at room temp for two months
without affecting its flavor
Chemical Methods of
Disinfection
 Alcohols
 Aldehydes
 Halogens
 Heavy metals
 Quaternary ammonium compounds
 Phenolics
Chemical safety
Material Safety Data Sheet

 Substance name
 Name, address, and telephone number of manufacturer
 Hazardous ingredients
 Physical and chemical properties
 Fire and explosion data
 Toxicity
 Health effects and first aid
 Stability and reactivity
 Shipping data
 Spill, leak, and disposal procedures
 Personal protective equipment
 Handling and storage
 Chemical spills:
 1st aid: flush the area with large amounts of water for at
least 15 minutes
 Do not neutralize chemicals
 Chemical handling:
 Always acid to water
#YouWereBornRight
 Yellow: reactivity or stability
hazard
 White: specific hazard
 Blue: Health hazard
 Red: Flammability hazard
Fire saftey
When a fire is discovered

#RACE

R – Rescue Rescue everyone in immediate

danger
A – Alarm Activate the fire alarm
C – Contain Close all the doors to
potentially affected areas
E – Extinguish Attempt to extinguish the fire
To operate a fire extinguisher

#PASS

P – Pull the pin

A – Aim at the base of fire
S – Squeeze handles
S – Sweep nozzle side to side
Electrical hazards
 Do not operate equipment with wet hands
 All electrical equipment must be grounded with three-
pronged plugs
 If electrical shock occurs, never touch the person or the
equipment involved
 Turn off the circuit breaker
 Unplug the equipment
 Move the equipment using a nonconductor object
 HANDLING OF COMPRESSED GASES
 BIOSAFETY
 EXPOSURE CONTROL PLAN
 EMPLOYEE EDUCATION AND ORIENTATION
 DISPOSAL OF HAZARDOUS WASTE
 STANDARD PRECAUTIONS
 ENGINEERING CONTROLS
BIOLOGIC SAFETY CABINET

 device that encloses a workspace in such a way as to

protect workers from aerosol exposure to infectious
disease agents
 type of containment barrier that protects the worker
from the aerosolized transmission of organisms.
 Class I Cabinets: Allow room (unsterilized air) to pass
to the cabinet
 Sterilize only the air to be exhausted
 Class II Cabinets: Laminar flow
 Sterilize air that flows + air exhausted
 Class III Cabinets: highest safety
 Air in and out is sterilized (HEPA)
 Material is handled with rubber gloves attached and sealed
to the cabinet
CLASSIFICATION OF BIOLOGIC
AGENTS
BASED ON HAZARD
 BSL-1
 BSL-2
 BSL-3
 BSL-4