STERILISATION AND DISINFECTION

-BHOOMIKA S
III RD BDS
CONTENTS
 INTRODUCTION
 TERMINOLOGIES
 HISTORY OF STERILISATION
 CLASSIFICATION OF STERILISING AGENT
 PHYSICAL AGENTS
 CHEMICAL AGENTS
 DISINFECTANTS
 BIOLOGICAL CONTROL
 STERILISATION IN DENTISTRY
 INFECTION CONTROL
 WASTE MANAGEMENT
 RECENT ADVANCES
 CONCLUSION
 REFERENCES
 INTRODUCTION
• Microorganisms are found in surroundings, in inanimate
objects and on surface of human body it causes
contamination, infection and decay .It becomes necessary to
remove or destroy them.
• In spite of scientific advances in understanding pathogenesis
of various dental diseases. Infection remain as antagonists for
surgeons.
• So, every attempt is made to minimize infection rates in
wards, hospital and operation theatre .
TERMINOLOGIES
• STERILISATION-process by which an article ,surface or medium is freed of all living
organisms either in vegetative or spore state.
• DISINFECTION- destruction of all pathogen organism by physical or chemical means.
• ANTISEPTICS-agents that can safely applied on the skin or mucous membrane to
prevent infection by inhibiting growth of bacteria.
• BACTERICIDAL-substances that can kill bacteria.
• BACTERIOSTATIC-prevent multiplication of bacteria .
• DECONTAMINATION –process of rendering an article area free of contaminants
including microbial, chemical ,radioactive and other hazardous material free from
an area .
HISTORY OF STERILISATION
• HIPPOCRATES OF COS (460BC) disproved the idea that disease was
punishment of sin. He advocated irrigation of wounds with wine or
boiled water.

• IGNAS ,an Hungarian obstetrician ,advocated in 1847 the value of hand

washing and fingernail scrubbing.
• LOUIS PASTEUR in 1862 ,French chemist and microbiologists publishes his
finding on how germs cause disease, later uses to develop pasteurization
process.

• JOSEPH LISTER, an English physician, uses a carbolic solution spray as he

operated ,he then used in wound.

• CHARLES CHAMBERLAND, developed the first pressure sterilizer or

autoclave in 1876
• ROBERT KOCH and his associates in 1881 on the
disinfecting properties of steam and hot air mark
the beginning of science of disinfection and
sterilization .

• AESCULAP, created the first rigid container. made

of stainless steel. He manufactured chrome plated
container for safe transport of sterile instruments.
CLASSIFICATION OF STERILISING
AGENTS
 CHEMICAL AGENTS PHYSICAL AGENTS

ALCOHOLS: ETHYL, ISOPROPYL .

DRY HEAT : FLAMING, INCERINATION
ALDEHYDES : FORMALDEHYDE
AND HOT AIR
GLUTARALDEHYDE.
MOIST HEAT : BOILING
ORTHOPHTALDEHYDE
STEAM AT ATMOSPHERIC PRESSURE
PARACETIC ACID
 STEAM ABOVE
HYDROGEN PEROXIDE
ATMOSPHERIC
HYPOCHLOROUS ACID
PRESSURE
DYES
HALOGENS
FILTRATION :CANDLES
PHENOLS
 ASBESTOS
SURFACE ACTIVE AGENTS
 MEMRANES
METALLIC SALTS
RADIATION
GASES:ETHYLENE OXIDE
,BETAPROPIOLACTONE
 PHYSICAL AGENTS
• HEAT DRY HEAT MOIST HEAT

FACTORS INFLUENCING

N
TIO
STERILISATION BY HEAT ARE PROTEIN DENATURATI
NATURE OF HEAT

AC
DENATURATI ON AND

OF
TEMPERATURE AND TIME ON COAGULATIO

SM
NUMBER OF N OF

NI
MICROORGANISMS PROTEINS
HA
TYPE OF MATERIAL FROM EC
M
WHICH ORGANISM ERADICATED.
 DRY HEAT
• FLAMING : Holding instruments over a bunsen flame till
they become red hot .
• Inoculating loop dipped in disinfectant before flaming
• E.g : INOCULATING LOOP OR WIRE,TIP OF FORCEPS
AND SEARING SPATULAS.
 INCINERATION
• This is an excellent method for terminal sterilization for
destroying biomedical waste .
• But, polystyrene emit clouds of dense smoke that pollutes
environment .
• E.g :plastics such as PVC and polyethylene.
DESIGN AND
CONSTRUCTI
ON OF
BIOMEDICAL
WASTE
INCINERATO
R
 HOT AIR OVEN
• This Is most widely used method by dry heat.
• Principle: BY conduction.
• Temperature: 160 c for 2 hours .
• Materials sterilized- Glassware
,forceps
Scissors and
scalpels
All glass
syringes, swabs

Oils ,fats, grease

• Precautions
• Not to be overloaded.
• Must be fitted with fan for even distribution of hot air.
• Materials to be sterilized to be dry .
• Rubber materials will not withstand the temperature.
• Allowed to cool for 2 hrs before opening .

ADVANTAGES DISADVANTAGES

ECONOMICAL IT HAS LESS PENERATING

POWER.

DOES NOT RUST METALS

EASILY MONITORED
 GLASS BEAD AND HOT SALT STERILISER

• This method employs a heat transfer device.

• Media used are glass beads, molten metal and salt.
• Temperature achieved is of 220 °c .
• Submersion of small instrument such as endodontic files, burs sterilised in
10 seconds.
• A warm up time of 20 min is atleast given for ensuring uniform heating.
 MOIST HEAT

TEMPERATURE BELOW TEMPERATURE AT 100°

C TEMPERATURE ABOVE
100° C Boiling
Pasteurization of 100 °C
milk. Tyndallisation Autoclave
Inspissations Steam sterilization
at 100
 PASTEURISATION OF MILK

 The milk heated at 63 °c for 30 minutes (holder method).

 Heated at 72°c for 15-20 seconds (flash process).
 And sudden cooling to 13 °c or lower.
 These process helps in elimination of all non sporing pathogens such as
mycobacterium ,brucella and salmonellae .
 Coxiella brucella is severely heat resistant and may survive .
 INSPISSATOR

• The materials are rendered sterile by heating at 80-85 °c for half an

hour on 3 successive days in inspissator.
• E.G: Lowenstein-Jensen and Loffler serum .

Bacterial vaccines Serum or body

inactivated at fluids for 1 hr at
60°c for 1 hr 56° c

ALL NON SPORING Streptococcus feocalis

and staph aureus
KILLED AT 60 °C killed at 60 °c for 60
IN 30 MIN min
 BOILING
• Vegetative bacteria killed almost immediately at 90-
100°c (spores withstand temperature).
• In case ,where boiling is adequate immerse it in water for 10 to 30 min.
• Not recommended for instruments used in surgical procedure.
• Hard water not used for boiling.
• Instead, addition of 2 % sodium bicarbonate is added to render it soft.
 STEAM AT ATMOSPHERIC PRESSURE

 An atmosphere of free steam is used to sterilise culture media (decompose if subject to high
temperature).
 Also called KOCH OR ARNOID STEAMER.
 TYNDALLISATION OR INTERMITTENT STERILISATION

 Method used for media containing sugar or gelatin.

 An exposure to 100 °c for 20 minutes on 3 successive days used.
 PRINCIPLE:
 First exposure kills all vegetative bacteria and spores .Since, they are ain favourable medium ,will germinate and
killed on subsequent exposure to 100°c for 20 m,inutes.

 However, fails to kill spore of certain species ,especially thermophiles.

 AUTOCLAVE
• PRINCIPLE: “WHEN WATER BOILS ,ITS VAPOUR PRESSURE
IS EQUAL TO THAT OF SURROUNDING ATMOSPHERE.”
• As pressure inside closed vessel increases, temperature at
which water boils also increases forming steam.
• Steam contact cooler surface condenses latent heat .This
process continues until temperature of fluid inside is
similar to that of steam outside.
• TEMPERATURE: 121°C ,TIME : 15 MIN ,PRESSURE :15
PSI
• PRESSURE TEMP TIME
(PSI) (°C) (min)
MATERIALS STERLISED
15 121 15
LINEN,INSTRUMENTS,LA
20 126 10 B WARE,MEDIA AND
20 134 3 PHARMACEUTICAL
PRODUCTS,AQUEOUS
SOLUTIONS.
• AUTOCLAVES USED IN HEALTH CARE SETUP INCLUDE
• lab autoclave
• Hospital dressing sterilizers
• Instrument steriliser
• Rapid cooling sterilizers.
CLASSIFICATION SUITABLE FOR USED BY
PROCESSING
DOWNWARD Unwrapped soild
DISPLACEMENT (N instruments for
TYPE) immediate use
VACCUM TYPE (S) Items specified by Medical surgeries,
autoclave pediatrist
manufacturer. Used
for single wrapped
soild and hollow
items

B TYPE VACCUM Wrapped and Dentists, plastic

unwrapped solid surgeons.
instruments
 TYPES OF AUTOCLAVE
• DOWNWARD DISPLACEMENT (GRAVITY DISPLACEMENT).
• POSITIVE PRESSURE DISPLACEMENT.
• NEGATIVE PRESSURE DISPLACEMENT (MOST ACCURATE).
• TRIPLE VACCUM AUTOCLAVE .
.
PRECAUTIONS DURING
AUTOCLAVING
ENSURE COMPLETE AIR REMOVAL FOR 121°C
●
INSTRUMENT STORED IN DAMP CONDITION LEADS TO TARNISH AND COROSSION

ENSURE LOOSE PACKING IN CHAMBER

●
TIGHTLY SEALED MATERIALS MAY BECOME DANGEROUSLY PRESSURISED CAUSING INJURY IF REMOVED
• ADVANTAGE • DISADVANTAGE
 ECONOMICAL i. MOISTURE RETENTION
 GOOD PENETRATION ii. CAUSES CORROSION
 SHORT CYCLE TIME iii. CARBON STEEL
 EASY MONITOR DAMAGED
iv. DESTRUCTION OF HEAT
 NO SPECIAL CHEMICAL
SENSITIVE MATERIALS
USED
FILTRATION
• IT helps remove bacteria from heat labile liquids
such as sera and solution of sugars and antibiotic.
• Bacterial vaccines obtained by passing through
filters.
• TYPES OF FILTERS

CANDLE USED FOR

INDUSTRIAL AND
FILTERS DRINKING PROCESS
E.G :
DIATAMACEOUS
EARTH
SINTERED LOW ABSORPTIVE
PROPERTY AND
GLASS FILTERS CLEANED EASILY
MEMBRANE MADE OF
CELLULOSE ESTERS
FILTERS ANS USED FOR
STERILISATION AND
STERILITY TEST .
 RADIATION
NON IONISING IONISING GAMMA

USES LONGER •USES SHORTER •FORM OF PURE

WAVELENGTH AND WAVELENGTH AND ENERGY PENETRATE
LOW ENERGY HIGH ENERGY DEEP AND KILLS MICRO
ORGANISM
•INFRARED RAYS USED oTHIS RADIATION oBENEFITS
FOR RAPID MASS FORM OF X- RAYS oPRECISE DOSING
STERILISATION DAMAGE DNA

•E.G :SYRINGES AND NO APPRECIABLE RISE •RAPID PROCESSING

CATHETERS IN TEMPERATURE •UNIFORM DOSE
DISTRIBUTION ‘

•UV RAYS FOR COLD STERILISATION •DOSIMETRIC RELEASE

DISINFECTING CLOSED
AREAS LIKE OT AND
LABS
 CHEMICAL AGENTS
•ACTION BY
•PROTEIN COAGULATION
•DISRUPTION OF CELL MEMBRANE
•REMOVAL OF FREE SULPHADRYL MOLECULE
•SUBSTRATE COMPETITION.
 CLASSIFICATION OF INSTRUMENTS

SEMI CRITICAL NON CRITICAL

CRITICAL INSTRUMENTS
INSTRUMENTS INSTRUMENTS

PENETRATE SOFT CONTACT THE

TISSUE MUCOUS MEMBRANE
COME IN CONTACT
CONTACTS BONE BUT WILL NOT
WITH INTACT SKIN
PENETRATE SOFT
TISSUE

IN CONTACT WITH E.G:MOUTH

E.G : X RAYS, LIGHT
BLOOD .E.G : SURGICAL MIRRORS,IMPRESSION
SCISSORS,DENTAL BURS,
HANDLES,COUNTER
TRAYS,PROBE,TWEEZE
FORCEPS, BONE GRAFT TOPS
RS
 ALCOHOLS
• Ethyl or isopropyl alcohol are most frequently used
• Mainly as skin antiseptics at conc. of 60 to 90 % in water.
• Isopropyl alcohol is more bactericidal and less volatile.
• Methyl alcohol effective against fungal; spores used for cleaning
cabinets and incubators .
 AQUEOUS QUATERNARY AMMONIUM COMPOUNDS

• Benzalkonium chloride (zephiran) commonly used as antiseptic

and disinfectant.
• It is reported that spectrumn of activity is limitedly largely to gram
positive and some gram negative.
• It is strongly surfactant that increases bacterial permeablity of
bacterial cell wall and escape of pottasium and nitrogen .
 ALDEHYDES

• FORMALDEHYDE
• It is markedly bactericidal, sporicidaland virucidal.
•To preserve anatomical specimens
•10% formalin containing 0.5% sodium
tetraborate used to sterilise clean
instrument
•To sterilise heat sensitive catheters
and fumigation of wards

•Under properly controlled condition

•Clothing, bedding,furniture is
disinfected .
• GLUTARLDEHYDE:Action especially against tubercle, fungi and viruses.
used to sterilise cystoscopes,bronchoscopes,rubber
anesthetic tube,endotracheal tubes and metal instrument.

• ORTHOPHTHALDEHYDE:It has bactericidal activity.

used to cleanse endoscopes between patient
as it is effective.
• PARACETIC ACID:Has good sterilisation effect on bacteria,
particularly antibiotic resistant bacteria such as methicillin resistant
staphylococcus aureus,vancomycin resistant enterococcus and clostridium
dificile.

• HYPOCHLOROUS ACID:ACTIVE AGAINST BIOFILMS AND MICRO

ORGANISM WITH BIOFILMS.
 DYES

• TWO GROUPS OF DYES

• Aniline dye and Acridine dye extensively used as skin and
wound antiseptics.
• Both are bacteriostatic in high dilution but low bactericidal
activity.
• More active against gram positive.
 HALOGENS

• IODINE •USED AS SKIN DISINFECTANT

•BACTERICIDAL AND MODERATE ACTION
AGAINST SPORES
•ACTIVE AGAINST TUBERCLE BACILI
•IODOPHORS HAVE BETTER ACTION .

• CHLORINE : •THEY ARE MARKEDLY BACTERICIDAL AND

VIRUCIDAL.
•AS DISINFECTANT IN POOLS ,DAIRY
SOURCES.
•CHLORAMINE IN SURGICAL DRESSING .
 PHENOLS
• WIDELY USED AS DISINFECTANTS IN HOSPITAL.
• COMMONLY USED AS LYSOL AND CRESOL.
• CHLOROHEXIDINE IS A RELATIVELY NON TOXIC ,SKIN
ANTISEPTIC AND WOUND DRESSING .
• CHLOROPHENOLS OR CHLOROXYPHENOLS ARE LESS ACTIVE
FORM OF PHENOL .
 GASES

• ETHYLENE OXIDE :
• It is a colourless liquid with a boiling point of 10.7°c.
• It is highly inflammable and explosive at concentration more than 3 %
• In combination with 10 % carbon dioxide it is explosive.
• It acts by alkylating amino,carboxyl,hydroxyl,and sulphydryl groups in protein molecule.
• Diffuses through many types of porous material and readily penetrates plastic.
•Heart lung machines
•Respirators
•Suture materials
•Dental equipment
•Books
•Clothing
• FORMALDEHYDE:THIS IS EMPLOYED FOR FUMIGATION OF
OPERATION THEATRE AND OTHER ROOMS.
• FORMALDEHYDE GAS IS GENERATED BY ADDING 150 gm of Kmno4 to 280 ml
formalin.
• Sealed room is left unoperated for 48 hrs after fumigation.
• BETAPROPRIOLACTONE:THIS IS A CONDENSATION PRODUCT OF
KETANE AND FORMALDEHYDE.
• NO LONGER USED AS IT IS CARCINOGENIC.
 DISINFECTANTS

BORIC ACID : USED AS

ANTISEPTIC FOR EYES AND
WEAK DISINFECTANT. • OXIDISING AGENTS
MUCOUS MEMBRANE. • HYDROGEN PEROXIDE AND
ETHYLENE OXIDE.
•SALTS OF HEAVY METALS:AS •LOWER TEMPERATURE GAS
•OZONE STERLIZATION
•DETERGENTS:SURFACE
ACTIVE AGENTS

•BETA PROPIOLACTONE
 LEVEL OF DISINFECTION
LEVEL OF ITEMS TIME DISINFECTANT
DISINFECTION

HIGH LEVEL CRITICAL AND SEMI >=20 MIN, GLUTALDEHYDE,HYD

CRITICAL ROGEN
ITEMS(EXCEPT PEROXIDE,PARACETI
THERMOMETERS ) C ACID.

INTERMEDIATE SEMICRITICAL AND <=10 MIN ALCOHOLS,IODOPHO

LEVEL NON CRITICAL RS,PHENOLS,CHLORI
(EXCEPT NS.
ENVIRONMENTAL
SURFACES)

LOW NON CRITICAL ALCOHOL,IODOPHO

LEVEL(ENVIRONMEN RS,PHENOLICS.
TAL SURFACE)
 BIOLOGICAL CONTROL

THERMOCOUPLE
THERMOMETRIC TESTING .
Recording is taken from
thermocouple placed in
test pack pf towels and one
from chamber. Speed at
which steam penetrate.
AUTOCLAVE TAPE
Tape printed with specific
ink and undergo change at
particular temperature.
Forms basis of bowie-dick
test for high vacuum
sterilizers.

SPORES OF NON
PATHOGENIC
ORGANISM BROWNS TEST
Organism used is an Ampoules that
aerobic spore of Bacillus contain chemicals.
thermophilus.this spore Changes from red
are killed at 121°c after 15 through amber to
min and couture green
 STERILISATION IN DENTISTRY

• THE FOUR ACCEPTED METHOD OF STERILISATION IN

DENTAL OFFICE
• STEAM PRESSURE STERILISATION
• CHEMICAL VAPOUR STERILISATION
• DRY HEAT STERILISATION
• ETHYLENE OXIDE STERILISATION
 STERILISATION OF MOST COMMONLY USED MATERIALS IN DENTISTRY

• DENTAL RADIOGRAPHY:CONTAMINATION OCCURS MOSTLY FROM SALIVA

• PRECAUTION TO BE TAKEN TO PLACE FILM PACKETS AND FILM HOLDERS IN SPECIAL TRAY.
• FILM HOLDING DEVICE TO BE RINSED IN RUNNING WATER TO REMOVE SALIVA.
• WIPE THE X RAY TUBE HEAD,EXPOSURE SELECTOR,TIME BUTTON WITH DETERGDENTS.

• BIOPSY SPECIMEN:BIOPSY COLLECTION AND TRANSPORTATION CAN BE A

SOURCE OF INFRCTION.
• KEPT IN STURDY ENVIRONMENT WITH SEVERE LID.

• IMPRESSION TRAY: THEY ARE STERLILISED USING AUTOCLAVE FOR METALS.

• ETHYLENE OXIDE FOR PLASTIC TRAY.
 BIOFILMS

• Tubes containing hand pieces, air /water syringe and

ultrasonic scalers unit harbor a wide range of
microorganism.
• Measures taken are :
• Anti- retraction valves.
• Bacterial filter
• Chemical disinfection.
• Aspirators.
• IMPLANTS : PRE STERILISED WITH GAMMA RADIATION .
• STEAM STERILISERS ARE NOT USED AS IT RESULTS IN CONTAMINATION OF SURFACE
WITH ORGANIC SUBSTANCES.
• RADIO FREQUENCY GLOW DISCHARGE OR PLASMA STERLISING USED.

• SUTURES :PRE STERILISED BY GAMMA RADIATION .

• RESTERILISED BY
• SOAK FOR COMPLETE 10 MIN COMPLETELY IMMERSED IN PROVIDONE IODINE 10%
THEN RINSE WITRH WATER.
• ETHYLENE OXIDE –STEAM STERILISATION
 BURS

• DIAMOND AND CARBIDE BUR

• AFTER USE THEY ARE PLACED IN 0.2% GLUTARLDEHYDE AND SODIUM
PHENATE(SPORICIDIN)FOR AT LEAST 10 MINUTES.
• STERILISE IN ULTRASONIC BATH.

• STEEL BURS
• MAY GET DAMAGED BY AUTOCLAVING
• CAN BE STERILISED BY CHEMICAL VAPOR STERILISEROR GLASS BEAD STERILISER AT 230°C FOR
20-30 SECONDS.
INFECTION CONTROL
 BASIC CONCEPT OF INFECTION CONTROL
• PREVENT SPREAD OF INFECTION FROM CLINICIAN TO PATIENT .
• PREVENT SPREAD OF INFECTION FROM PATIENT TO CLINICIAN .
• PREVENT SPREAD OF INFECTION FROM ONE PATIENT TO
ANOTHER.
 OPERATING ROOM PROCEDURES

• The ceiling,walls and floors are regularly disinfected

after contaminated site.
• The access to operation theatre is done with special
scrub dress.
• A surgical cap to tie hair completely .
• Mask is then placed and tied over nose and mouth .
• Conductive shoes are usually used as they build up
static electricity.
• Operation theatres are fumigated with use of
pottasium permagnate reaction technique.
• The chemical used is 40 % formaline and fumigator
set for 30 min.
• EYE WEAR :
• It is advisable to wear eye wear protection during
any procedure that is likely to generate droplets of
blood .
• Protect mucous membrane of eye from blood
borne diseases.
 Foot wear
• Staff membrane should wear clean,comfortable
and antistatic shoes.
• Footwear should avoid producing bellows effect .
• Construction to be robust to protect feet from
sharp injury .
HAND DISINFECTION
WEARING GLOVES
WASTE MANAGEMENT
 RECENT ADVANCES IN STERILISATION
• PEROXIDE VAPOUR STERILISATION :An aquaeous
hydrogen peroxide solution boils in heated vapour
and flows as vapour in sterilisation chamber.
• PLASMA STERILISATION: Using photolytic,
chemical, and electric action effectively kills micro
organisms.
• OZONE STERILISATION : it is newest method and
most compatible with stainless steel .
• Cycle approximately for 4.5hrs at temp 85°f.
 NEWER DISINFECTANTS

• Persistant antimicrobial drug coating applied on

inamniate object (surfacine)
• A high level disinfectant with reduced exposure time
(orthopthaldehyde).
• Superoxide water.
 CONCLUSION
ARTICLES STERILISED METHOD OF STERILISATION OR
DISINFECTION

• DISPOSABLE SYRINGES GAMMA RADIATION AND ETHYLENE OXIDE.

• GLASS SYRINGES (NON DISPOSABLES) BOILING AT 100 °C,AUTOCLAVE,HOT AIR

OVEN.

• SURGICAL NEEDLES HOT AIR OVEN ,AUTOCLAVE

• CYTO SCOPE, ENDOSCOPE GLUTARALDEHYDE,ETHYLENE OXIDE.

• HEART LUNG MACHINE ETHYLENE OXIDE

• APRONS,GLOVES, CATHETERS AUTOCLAVING

• DRESSING,LINEN (GOWNS) AUTOCLAVING

• CLINICAL THERMOMETER,STETHOSCOPE SOAP WATER/ETHYL ALCOHOL AT 70 %

• OPERATION THEATRE FORMALDEHYDE, UV RAYS

•CHITTEL FORCEPS SAVLON

•SHARP INSTRUMENTS 5%CRESOL

•SPUTUM AUTOCLAVE, INCERINATION ,BURNING

•CULTURE MEDIA AUTOCLAVE

•BLOOD BAGS MICROWAVE. INCINERATOR

•THROAT SWAB HOT AIR OVEN

•ANTIBIOTICS,SERA FILTRATION

•DISINFECTION OF SURFACE HYPOCHLORITE

•METAL INOCULATING WIRE READ HEAT IN BUNSEN BURNER

•OILS, POWDERS HOT AIR OVEN