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Asepsis, Sterilizationand Disinfection
Pradeep B Patil
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Sushruta 1500BCE
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Hippocrates (460-377 BC)
Separated medicine from
philosophy
Irrigated wounds with wineor boiled water
Foreshadowing ofAsepsis
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Joseph Lister
chemical antiseptics tocontrol surgery relatedinfections
disinfect surgical equipmentand supplies
Microorganisms are simple forms of life commonly made up
of a single cell only seen with a microscope.
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Pasteur (1862)
Discovered bacterium
Developed Pasteurizationprocess
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Mikulicz-Radecki (1887)---face masks
Furbringer (1889)-----------hands-scrubbingHalsted (1890)---------------surgical-gloves
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Basic terms
Sterilization
Disinfection
Antisepsis
Bacteriostasis
Asepsis
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Microbial Control
Degerming: Removing microbes from a limited area
Sanitization: Lowering microbial counts on eating
utensils
Biocide/germicide: Kills microbes
Bacteriostasis: Inhibiting, not killing, microbes
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Effectiveness of Treatment
Depends on
Number of microbes
Environment (organic matter, temperature, biofilms)
Time of exposure
Microbial characteristics
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Actions
Alteration of membrane permeability
Damage to proteins
Damage to nucleic acids
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1. Medical asepsis
Wash hands
Sanitize & disinfect equipment
2. Use of PPE
Gloves, Mask, Gown
3. Deal with waste & disposal
OSHA --- deals with waste in facility
EPA --- deals with waste outside facility
4. Sterilize instruments
5. Spillage
Standard Precautions
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Handwashing
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Using alcohol based hand rub
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SPILLS
should be promptly cleaned up
clean up = * gloved employee
* paper towels
* infectious waste container after clean up :
disinfect area with bleach solution
household bleach = 5.25% sodium hypochlorite
bleach solution = 1:10 dilution
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Surgical Asepsis
Surgical Asepsis and Sterilization
Steam Sterilization (Autoclave)
Dry Heat Sterilization
Gas Sterilization
Chemical Sterilization
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1.Chemical agentsAlcohols,Chlorine, Formalin
Suitable for skin and instruments
2. Radiation
3. Dry Heat
Ultraviolet and Ionizing Radiation
Suitablefor sterile room and inoculation hood
Direct flaming: e.g. inoculating loop
Hot-air sterilization :160, 2h in hot air oven
4. Moist Heat12130min in autoclaveSuitable for medium and instruments
5. FiltrationRemoval of bacteria by filter medium
Used for heat sensitive materials and filtrated air
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Heat sterilizationhow it works?
Dry heat- protein oxidation
Moist heat- protein denaturation
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Wrapping for Sterilization Techniques involves sanitizing (washing & cleaning)
make sure things are in proper working order
these materials allow steam or chemical vapors to penetrate,
but do not allow airborne or surface contaminants to enter
muslin = cotton fabric----------------Shelf Life = 1 month
special paper ------------------------- Shelf Life = 1 month
special disposable envelopes ------ Shelf Life = 3 months
Technique for paper & muslin :
Diamond; Bottom; Right Side; Left Side ; Fold Up
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Flaming
Hot air oven
-170C for 1 hour
-140C for 3 hours
.
Dry Heat
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1. Pasteurization( below 100C)
Destroys pathogens without altering the
flavor of the food.
Low temperature (holding method): 63 , 30 min
High temperature (flash method) : 72 , 20sec
Moist Heat
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2. Boiling (at 100C)
-killing most vegetative forms of bacteria
-10 min or longer time
3. Autoclaving (above 100C)
-killing both vegetative organisms and endospores
-121-132 oC for 15 min or longer
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Moist Heat vs Dry Heat
Moist heat Dry heat
Penetrating potency higher lower
Temp. for protein clotting lower higher
Extra heat released yes no
from condensation
Sterilizing potency: Moist heatDry heat
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Why is moist heat more efficient than
dry heat ?
Conductivity. Moisture conducts the heat better than a
dry system.
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Sterilization processes
Batch sterilization
Continuous sterilization
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Filtration
Sterilize solutions that may be
damaged or denatured by high
temperatures or chemical
agents.
The pore size for filtering bacteria, yeasts, and
fungi is in the range of 0.22-0.45 m(filtration
membranes are most popular for this purpose).
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Air Sterilization
Very large volumes of sterile air is required in many aerobic
fermentation process.
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Methods for air sterilization
Radiation
High temperature
Electrostatic bacteria removal
Filtration
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WASTES1. Medical 2. Contaminated 3. Non-Medical
Medical ( Infectious) Waste 1. laboratory waste = chemicals, detergents, containers
2. pathology specimens = human tissue, blood, excrement, secretions
3. surgical specimens = human parts removed at surgery / autopsy 4. sharps = any device having rigid areas capable of cutting / piercing
5. liquid body waste = stool, excretions, secretions
6. contaminated waste things that have been used on body (dressings)
Non-Medical Waste - non-patient care waste - kitchen waste
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Aseptic Technique
1. Remove all jewelry (e.g., watches, rings, bracelets,
necklaces.
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2. Put on non-shedding coats, gowns, or coveralls (hospital
scrubs);
head and facial hair covers; face masks; and shoe covers.
Note that it is important to follow the sequence of items
indicated in this step.
Aseptic Technique
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Personnel protective equipment
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Keyboard and mouse with proper barrier protection
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3. Scrub hands and arms to the elbows thoroughly with
an antiseptic cleanser
(e.g., povidone-iodine or chlorhexidine gluconate).
Aseptic Technique
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4. Clean the laminar flow hood with isopropyl alcohol.
The alcohol must remain in contact with the surface for
30 seconds prior to compounding any sterile product.
Aseptic Technique
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5. Place only essential materials under the airflow hood
no paper, pens, or labels.
Remove the selected syringe(s) from its overwrap, attach aneedle, then discard the waste.
Aseptic Technique
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6. Scrub again and glove.
Aseptic Technique
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7. Swab or spray needle-penetration closures on vials,
injection ports, and other materials.
Aseptic Technique
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8. Prepare the sterile product by
withdrawing the medication from vials or ampules
introducing it into the IV container.
Aseptic Technique
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9. Complete a quality check of the product for
container integrity and leaks,
solution cloudiness,
particulates,
color of solution
proper preparation of product.
Aseptic Technique
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Figure 10.2
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HANDLING AND DISPOSAL OF
HAZARDOUS AGENTS Receipt and Storage of Hazardous Agents:
Hazardous drugs should be delivered directly to the
storage area, inventoried, and, if necessary, refrigerated.
Access to storage areas and work areas for hazardous
materials should be limited to specified trained personnel.
S# Waste class Type of container Colour
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S# Waste class Type of container Colour
1. Human anatomicalwaste, Animal wasteSolid (biomedicalwaste)
Plastic Yellow
2. Microbiology andBiotechnology waste
Plastic Red
3. Waste sharp Plastic bagpuncture proof
containers
Blue/WhiteTranslucent
4. Discarded medicinesand Cytotoxic waste
Chemical waste(solid)Incineration waste
Plastic bag Black
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Reporting Guidelines
National Notifiable Disease Surveillance System
Examples
HIV/AIDS Cholera
Rubella
Hepatitis Legionellosis Lyme disease
Mumps Smallpox
Tetanus Tuberculosis
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Sterilization of Surgical Instruments
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1906
1933
1947Steam Sterilization
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Early type of hot air sterilizer
Microwave Oven (1947)
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Advanced Sterilizer
Low temperature system for endoscopic devices
(1989)
Plasma sterilizing system
(1993)
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Steam Sterilization
Steam must contact items surface
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Filtration
HEPA removes
microbes >0.3 m
Membrane filtration
removes microbes
>0.22 m
Ph i l M th d f Mi bi l
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Physical Methods of Microbial
Control
Low temperature inhibits microbial growth
Refrigeration
Deep-freezing
Lyophilization
High pressure denatures proteins
Desiccation prevents metabolism
Osmotic pressure causes plasmolysis
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Figure 7.5
Radiation
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Radiation
Ionizing radiation (X rays, gamma rays, electron
beams)
Ionizes water to release OH
Damages DNA
Nonionizing radiation (UV, 260 nm)
Damages DNA
Microwaves kill by heat; not especially antimicrobial
Principles of Effective
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Principles of Effective
Disinfection
Concentration of disinfectant
Organic matter
pH
Time
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Biguanides
Chlorhexidine
Disrupt plasma membranes
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Halogens
Iodine
Tinctures: In aqueous alcohol
Iodophors: In organic molecules
Alter protein synthesis and membranes
Chlorine
Bleach: Hypochlorous acid (HOCl)
Chloramine: Chlorine + ammonia
Oxidizing agents
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Table 7.6
Alcohols
Ethanol, isopropanol
Denature proteins, dissolve lipids
Require water
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Heavy Metals
Ag, Hg, and Cu
Silver nitrate may be used to prevent
gonorrheal ophthalmia neonatorum
Silver sulfadiazine used as a topical cream onburns
Copper sulfate is an algicide
Oligodynamic action Denature proteins
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Chemical Food Preservatives
Organic acids
Inhibit metabolism
Sorbic acid, benzoic acid, and calcium propionate
Control molds and bacteria in foods andcosmetics
Nitrite prevents endospore germination
Antibiotics Nisin and natamycin prevent spoilage of cheese
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Aldehydes
Inactivate proteins by cross-linking with
functional groups (NH2,OH,COOH,
SH)
Use: Medical equipment
Glutaraldehyde, formaldehyde, and ortho-
phthalaldehyde
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Gaseous Sterilants
Denature proteins
Use: Heat-sensitive material
Ethylene oxide
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Infrared rays
Temperature =160
Time = 60-120 min
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Supercritical Fluids
CO2 with gaseous and liquid properties
Use: Medical implants
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Peroxygens
Oxidizing agents
Use: Contaminated surfaces
O3, H2O2, peracetic acid
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Figure 7.11
Microbial Characteristics
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Table 7.7
Endospores and Mycobacteria
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Disposal container for contaminated sharps
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