THIRD YEAR

B.Sc Operation Theatre and anaesthesia technology

STERILIZATION PROCEDURES

Bio medical waste :

 Biomedical waste is any kind of waste containing infectious (or potentially infectious)
materials.It may also include waste associated with the generation of biomedical
waste that visually appears to be of medical or laboratory origin (e.g., packaging,
unused bandages, infusion kits, etc.), as well research laboratory waste containing
biomolecules or organisms that are mainly restricted from environmental release.
 As detailed below, discarded sharps are considered biomedical waste whether they
are contaminated or not, due to the possibility of being contaminated with blood and
their propensity to cause injury when not properly contained and disposed of.
 Biomedical waste may be solid or liquid. Examples of infectious waste include
discarded blood, sharps, unwanted microbiological cultures and stocks, identifiable
body parts (including those as a result of amputation), other human tissue,
used bandages and dressings, discarded gloves, other medical supplies that may have
been in contact with blood and body fluids, and laboratory waste that exhibits the
characteristics described above.
 Waste sharps include potentially contaminated used (and unused
discarded) needles, scalpels, lancets and other devices capable of
penetrating skin.
 Biomedical waste is distinct from normal trash or general waste, and differs from
other types of hazardous waste, such as chemical, radioactive, universal or industrial
waste. Medical facilities generate waste hazardous chemicals and radioactive
materials. While such wastes are normally not infectious, they require proper disposal.
Some wastes are considered multihazardous, such as tissue samples preserved
in formalin.
Four types of Biomedical Waste :
General Medical Waste
General medical waste is the lion’s share of medical waste in a facility and is
not typically considered hazardous. This includes paper, plastic, and office waste. These can
disposed of regularly and don’t require any special handling.

Infectious Medical Waste

Infectious waste is just what the name suggests: it is waste materials that
can pose a risk of infection to humans, animals, and the overall environment. This includes
blood-soaked bandages, sharps waste, surgical waste, human or body parts, cultures, and
swabs. Each state has comprehensive rules for the management of infectious waste,
including requirements for storage, transport, disposal, licensing, and processing.

Hazardous Medical Waste

Hazardous waste is dangerous waste but is not considered infectious to
humans. Believe it or not, sharps falls into this category as well, at least sharps that have not
been used, because they have the ability to puncture or harm the user. Chemotherapy agents
fall into this category, as well as chemicals, such as solvents, mercury in thermometers, and
lead in paint.

Radioactive Medical Waste

Simply put, radioactive waste is waste that contains radioactive material. In a
medical setting, this includes radioactive therapies for tests such as thallium stress tests, and
other nuclear medicine therapies to treat certain cancers. Nuclear medicine uses radiation to
provide diagnostic information about the functioning of a person’s specific organs, or to treat
them.

Medical waste must be collected by a licensed medical waste hauler, as this

waste disposal is closely monitored and regulated in most states. The waste must be treated
and rendered harmless before it can be recycled or thrown away.

Management and treatment :

 Biomedical waste must be properly managed and disposed of to protect the
environment, general public and workers, especially healthcare and sanitation
workers who are at risk of exposure to biomedical waste as an occupational hazard.
 Steps in the management of biomedical waste include generation, accumulation,
handling, storage, treatment, transport and disposal.
 The development and implementation of a national waste management policy
can improve biomedical waste management in health facilities in a country
 Disposal occurs off-site, at a location that is different from the site of generation.
Treatment may occur on-site or off-site. On-site treatment of large quantities of
biomedical waste usually requires the use of relatively expensive equipment, and is
generally only cost effective for very large hospitals and major universities who
have the space, labour and budget to operate such equipment.
 Off-site treatment and disposal involves hiring of a biomedical waste disposal
service (also called a truck service) whose employees are trained to collect and haul
-2-
away

-3-
 biomedical waste in special containers (usually cardboard boxes, or reusable plastic bins)
for treatment at a facility designed to handle biomedical waste
Storage :
Storage refers to keeping the waste until it is treated on-site or transported off-site
for treatment or disposal. There are many options and containers for storage. Regulatory
agencies may limit the time for which waste can remain in storage. Handling is the act of
moving biomedical waste between the point of generation, accumulation areas, storage
locations and on-site treatment facilities. Workers who handle biomedical waste must
observe standard precautions.
Treatment :
 The goals of biomedical waste treatment are to reduce or eliminate the waste's
hazards, and usually to make the waste unrecognizable. Treatment should render the
waste safe for subsequent handling and disposal. There are several treatment
methods that can accomplish these goals.
 Biomedical waste is often incinerated. An efficient incinerator will destroy
pathogens and sharps. Source materials are not recognizable in the resulting ash.
 An autoclave may also be used to treat biomedical waste. An autoclave uses steam
and pressure to sterilize the waste or reduce its microbiological load to a level at
which it may be safely disposed of. Many healthcare facilities routinely use an
autoclave to sterilize medical supplies.
 If the same autoclave is used to sterilize supplies and treat biomedical
waste, administrative controls must be used to prevent the waste operations from
contaminating the supplies.
 Effective administrative controls include operator training, strict procedures,
and separate times and space for processing biomedical waste.
 Microwave disinfection can also be employed for treatment of Biomedical wastes.
Microwave irradiation is a type of non-contact heating technologies for disinfection.
Microwave chemistry is based on efficient heating of materials by microwave
dielectric heating effects.
 When exposed to microwave frequencies, the dipoles of the water molecules present
in cells re-align with the applied electric field. As the field oscillates, the dipoles
attempts to realign itself with the alternating electric field and in this process, energy
is lost in the form of heat through molecular friction and dielectric loss.
 Microwave disinfection is a recently developed technology which provides
advantage over old existing technologies of autoclaves as microwave based
disinfection has less cycle time, power consumption and it requires minimal usage of
water and consumables as compared to autoclaves.

For liquids and small quantities, a 1–10% solution of bleach can be used to disinfect
biomedical waste. Solutions of sodium hydroxide and other chemical disinfectants may
also be used, depending on the waste's characteristics. Other treatment methods include
heat, alkaline digesters and the use of microwaves.
For autoclaves and microwave systems, a shredder may be used as a final treatment
step to render the waste unrecognizable. Some autoclaves have built in shredders.
-4-
Cleaning instruments :
I. Use stiff plastic cleaning brushes (nylon, etc.) Do not use steel wool or wire
brushes except specially recommended stainless steel wire brushes for instruments such as
bone files, or on stained areas in knurled handles.
II. Use only neutral PH(7) detergents because if not rinsed off properly, low PH
detergents will cause breakdown of stainless protective surface and black staining. High PH
detergent will cause surface deposit of brown stain, which will also interfere with smooth
operation of the instrument.
III. Brush delicate instruments carefully and, if possible, handle them totally
separate from general instruments.
IV. Make sure all instrument surfaces are visibly clean and free from stains and
tissue. This is a good time to inspect each instrument for proper function and condition.
Check and make sure that :
o Scissors blades glide smoothly all the way (they must not be loose when in
closed position). Test scissors by cutting into thin gauze. Three quarters of the
length of blade should cut all the way to the scissor tips, and not hang up.
o Forceps (pickups) have properly aligned tips.
o Hemostats and Needle Holders do not show light between the jaws, lock
and unlock easily, joints are not too loose.
o Check Needle Holders for wear on jaw surfaces.
o Suction tubes are clean inside.
o Retractors function properly.
o Cutting instruments and knives have sharp,
o undamaged blades.

V. After scrubbing, rinse instruments thoroughly under running water. While

rinsing, open and close Scissors, Hemostats, Needle Holders and other hinged instruments to
make sure the hinge areas are rinsed out, as well as the outside of the instruments.

1. RINSING
Immediately after surgery, rinse instruments under warm (not hot) running water.
Rinse should remove all blood, body fluids and tissue.

2. CLEANING
(If not done immediately after rinsing, instruments should be submerged in a solution
of water and neutral PH(7) detergent.)

A. Ultrasonic Cleaning
For micro and delicate instruments, use
manual cleaning (step C).
Instruments should be processed in a cleaner for the full recommended cycle time –
usually 5 to 10 minutes.
Place instruments in open position into the ultrasonic cleaner. Make sure that "Sharp"
(scissors, knives osteotomes, etc.) blades do not touch other instruments.
All Instruments have to be fully submerged.
Do not place dissimilar metals (stainless, copper, chrome plated, etc.) in the same
-5-
cleaning cycle.
Change solution frequently – at least as often as manufacturer recommends.
Rinse instruments after ultrasonic cleaning with water to remove ultrasonic
cleaning solution.

B. Automatic Washer Sterilizers

Follow manufacturers recommendations but make sure instruments are lubricated after
last rinse cycle and before sterilization cycle.

C. Manual Cleaning
Most instrument manufacturers recommend ultrasonic cleaning as the best and most
effective way to clean surgical instruments, particularly those with hinges, locks and
other moving parts. If ultra sonic cleaning is not available observe the following steps.

3. AFTER CLEANING
If instruments are to be stored, let them air dry and store them in a clean and dry
environment.

4. AUTOCLAVING
If instruments are to be reused or autoclaved:
A. Lubricate all instruments which have any "metal to metal" action such as scissors,
hemostats, needle holders, self-retaining retractors, etc. Recommend surgical lubricants such
as instrument milk are best. Do not use WD-40, oil or other industrial lubricants.
B. Put instruments up for autoclaving either individually or in sets.
Individual Instruments
Disposable paper or plastic pouches are ideal. Make sure you use a wide enough
pouch (4" or wider) for instruments with ratchet locks such as needle holders and
hemostats so the instrument can be sterilized in an open (unlocked) position.
Instrument Sets
Unlock all instruments and sterilize them in an open position. Place heavy instruments
on bottom of set (when two layers are required).
Never lock an instrument during autoclaving. It will not be sterile as steam cannot
reach the metal to metal surfaces. The instrument will develop cracks in hinge
areas because of heat expansion during the autoclave cycle.
Do not overload the autoclave chamber as pockets may form that do not permit steam
penetration. Place towel on bottom of pan to absorb excess moisture during autoclaving.
This will reduce the chances of getting "Wet packs". Make sure the towels used in
sterilization of instruments have no detergent residue and are neutral – PH(7) if immersed in
water.
This can be a real problem as laundries frequently use inexpensive but high PH(9-13)
detergents and do not properly rinse out or neutralize those detergents in the final
wash/rinse cycle.
Also, some times bleaches such as Clorox are added and are not neutralized.

-6-
CAUTION –
 At the end of the autoclave cycle – before the drying cycle – unlock autoclave door
and open it more than a crack (about 3/4"). Then run dry cycle for the period
recommended by the autoclave manufacturer.
 If the autoclave door is opened fully before the drying cycle, cold room air will rush
into the chamber, causing condensation on the instruments. This will result in water
stains on instruments and also cause wet packs.

5. COLD STERILIZATION
Most cold sterilization solutions render instruments sterile only after a 10 hour
immersion. This prolonged chemical action can be more detrimental to the surgical
instruments than the usual 20 minute autoclave cycle.
If the instruments need to be "disinfected" only, cold sterilization is okay as
disinfection will place in only 10 minutes.
But keep in mind the difference between:
STERILE – an absolute term (no living organism survives)
DISINFECTED – basically clean.
For instruments with Tungsten Carbide inserts (Needle Holders, Scissors, Tissue
Forceps), we do not recommend use of solutions containing Benzyl Ammonium Chloride
which will destroy the Tungsten Carbide Inserts.

Needle sterilization :
New needles that touch unsterile surfaces, such as a table or your hands, are no longer
sterile. Make sure to thoroughly wash your hands, and to use new surgical gloves before use.
Steam or boiling water is the best way to sterilize a needle you're planning to use for the
removal of a shallow splinter.

Newborn Care in the Hospital.

In the hospital, a routine evaluation called the Apgar test is used to identify whether
a baby needs urgent medical care. The test scores on five areas, including breathing rate and
skin coloration.Newborns also receive a vitamin K shot and eye drops

The hepatitis B vaccine protects against the hepatitis B virus, which causes liver
damage. The hepatitis B vaccine is a series of three shots. Your baby will usually receive
the first shot after delivery, and the next two shots by 18 months of age.

PRINCIPLES OF STERILIZATION & DISINFECTION. Sterilization is

the killing or removal of all microorganisms, including bacterial spores,
which are highly resistant.

sterilization refers to the set of operations that are developed to

eliminate or kill all forms of living beings that are contained in an
object or substance
Every heat-resistant material that is compatible with moisture should be
autoclaved. This is the principal method used in a SP
-7-
 Every heat-resistant material that is incompatible with moisture should
be sterilized by dry heat.
Sterilization with gaseous chemical methods should be carried out in
chambers with automated cycles that provide safety for the user and
guarantee the processes.
Sterilization by immersion in liquid chemical methods, which are
carried out manually, will always be the last method of choice.
These processes are difficult to control, have major possibilities of
recontamination during rinsing or drying, and do not allow for storage

Sterilization methods
o Physical methods: dry heat and moist heat.
o Chemical methods: liquids and gases (ethylene oxide).
o Physical-chemical methods: low-temperature steam
(formaldehyde) and gas plasma (hydrogen
peroxide).

MOIST HEAT :
Moist heat or steam sterilization describes sterilization techniques that uses hot air that
is heavily laden with water vapor

Description
Moist heat sterilization processes sterilize using hot air that is heavily laden with water
vapor, which plays the most important role in the sterilization. Boiling a sample for
30 minutes or more will kill virtually all vegetative cells present, but will not kill spores,
which can germinate shortly thereafter and resume growth. Therefore, boiling is an
insufficient method to achieve sterilization.

Action on micro-organisms :
Moist heat causes destruction of micro-organisms by denaturation of macromolecules,
primarily proteins. Destruction of cells by lysis may also play a role. While "sterility"
implies the destruction of free-living organisms which may grow within a sample,
sterilization does not necessarily entail destruction of infectious matter. Prionsare an example
of an infectious agent that can survive sterilization by moist heat, depending on conditions.

Validation
To facilitate efficient sterilization by steam and pressure, there are several methods of
verification and indication used; these include color-changing indicator tapes and
biological indicators.
When using biological indicators, samples containing spores of heat-resistant
microbes such as Geobacillus stearothermophilis are sterilized alongside a standard
load, and are then incubated in sterile media (often contained within the sample in a
glass ampule to be broken after sterilization).
-8-
 A color change in the media (indicating acid production by bacteria; requires the
medium to be formulated for this purpose), or the appearance of turbidity(cloudiness
indicating light scattering by bacterial cells) indicates that sterilization was not
achieved and the sterilization cycle may need revision or improvement.

Methods used
Tyndallization
A more effective method is Tyndallization, which uses three successive steam
treatments to achieve sterilization over the course of three days. This works by killing
vegetative cells, allowing germination of surviving spores, and killing the resulting
vegetative cells before they have time to form further spores. Any surviving spores from the
first treatment, or incidentally formed spores during the first incubation period, are killed in
a third steaming cycle.

High pressure
A more commonly used method when extended heat is not a concern is to use
an autoclave or pressure cooker. When sterilizing in this way, samples are placed into a
steam chamber on a shelf or raised floor, and the chamber is closed and heated so that steam
forces air out of the vents or exhausts. Pressure is then applied so that the interior
temperature reaches 121 °C (250 °F), and this temperature is maintained for between 15 and
30 minutes.
This elevated temperature and pressure is sufficient to sterilize samples of any
commonly encountered microbes or spores. The chamber is then allowed to cool slowly or
by passive heat dissipation; it is rare for forced cooling to be applied, or for pressure to be
vented deliberately. Pressure sterilization is the prevailing method used for medical
sterilization of heat-resistant tools, and for sterilization of materials for microbiology and
other fields calling for aseptic technique.
In cases when items need to be sterilized for immediate use, flash
sterilization may be employed. Flash techniques generally run for the minimum time,
temperature, or pressure, and may sacrifice some safeguards, such as the abilities to validate
with biological indicators or prevent contamination.Additional protocols are generally
taken to mitigate the sacrifices; flash sterilization equipment is often kept in an operating
room's sterile field, steam-penetrative protective packaging may be used to
prepackage items, and specially designed rigid sterilization container systems can be
reused.

Sterilizing agent:
Saturated steam at a pressure that is higher than the normal level.

Mechanism of action:
It acts by microbial death due to the denaturation of proteins, which is produced by the
action of the temperature and saturated steam. The temperature at which denaturation
occurs varies inversely with the amount of water present.

-9-
DRY HEAT STERILIZATION :
It uses hot air that is either free from water vapor or has very little of it, where this
moisture plays a minimal or no role in the process of sterilization.
Process
 The dry heat sterilization process is accomplished by conduction; that is where heat
is absorbed by the exterior surface of an item and then passed inward to the next
layer. Eventually, the entire item reaches the proper temperature needed to achieve
sterilization.
 The proper time and temperature for dry heat sterilization is 160 °C (320 °F) for 2
hours or 170 °C (340 °F) for 1 hour or in the case of High Velocity Hot Air sterilisers
190°C (375°F) for 6 to 12 minutes.
 Items should be dry before sterilization since water will interfere with the process.
Dry heat destroys microorganisms by causing denaturation of proteins.
 There are two types of hot air convection (Convection refers to the circulation
of heated air within the chamber of the oven) sterilizers:
 Gravity convection
 Mechanical convection

Mechanical convection process

A mechanical convection oven contains a blower that actively forces heated air
throughout all areas of the chamber. The flow created by the blower ensures uniform
temperatures and the equal transfer of heat throughout the load. For this reason, the
mechanical convection oven is the more efficient of the two processes.

High Velocity Hot Air :

An even more efficient system than convection uses deturbulized hot air forced
through a jet curtain at 3000ft/minute.

Instruments used for dry heat sterilization :

Instruments used for dry heat sterilization include hot air oven, incineration or
burning, flaming, radiation, microwave, bunsen burner, and glass bead sterilizer.

Effect on microorganisms :
Dry heat denatures the proteins in any organism, causes oxidative free radical
damage, causes drying of cells, and can even burn them to ashes, as in incineration.

Materials:
 Sharp stainless steel instruments (scissors and tweezers)
 Needles, crystal syringes, tubes, glass pipettes, heat-stable powders.
 Liquids and substances that are liposoluble and water-resistant such as
 oils, silicone, paraffin, vaseline, creams and talcum powders.

Sterilizing agent:
Hot air
Mechanism of action:
Microbial death occurs by causing denaturation of proteins due to presence of moisture.
- 10 -
EO STERILIZATION :
Ethylene oxide sterilization is a chemical process consisting of four primary
variables: gas concentration, humidity, temperature and time.
Mechanism of action:
EO is an alkylating agent that disrupts the DNA of microorganisms, which prevents
them from reproducing.
Stages of sterilization by EtO:
 Conditioning and humidification
 Entrance of the gas
 Exposure to the gas
 Evacuation
 Aeration
 Sterilization temperatures range from 35°C - 55°C and exposure times
range from 1 hour 20 minutes and 4 hours.
 The aeration process that should be implemented is carried out at 40°C - 60°C for 6 to
12 hours.
 This results in a total duration for the entire process of 8 to 16 hours.
 It is worth pointing out that implementation is carried out under the premise that lower
temperatures require longer aeration times
 Sterilization by EtO is recommendable provided that it is automated.

Advantages and disadvantages of the method:

Advantages:
EtO is a substance with a high level of diffusion and penetration, which permits
high versatility for the sterilization of heat-sensitive materials.

Disadvantages:
It is highly toxic to living things and can cause local reactions on skin and mucous
membranes and systemic toxic effects with clinical manifestations such as dyspnea,
- 11 -
 cyanosis, gastrointestinal disorders, hemolysis, necrosis, mutagenesis and
carcinogenesis.
Due to these adverse effects, it is considered a highly dangerous substance and its use
should be restricted to adequately trained personnel.
There are no chemical indicators that can monitor the concentration of Eto during the
sterilization cycle.
Symptoms associated with exposure to ethylene oxide:
Initially: irritation of the eyes, respiratory tract, nose and throat, with a "peculiar taste.
Late: headache, nausea, vomiting, dyspnea, cyanosis, pulmonary edema, weakness,
EKG abnormalities, urinary excretion of biliary pigments.
Skin irritation and burns through direct contact.
Elevated absolute white blood cell count and decline in hemoglobin values
The union of EtO produces a toxic compound called ethylene glycol, which depresses
the central nervous system and has renal toxicity.

Aeration:
The aeration of objects sterilized by EtO permits the desorption of the gas
Metal objects do not require aeration. However, the packaging used does.
The proposed aeration time for all materials is:

EtO Measurement :
Eight-hour controls should be carried out twice a year
Fifteen-minute controls should be carried out 4 times a year
This instrument or monitor, which looks like a dosimeter, should be placed as close as
possible to the operator's face, as if it were an "identification card”
Subsequent to exposure, the monitor should be sent for the corresponding
reading of the limit value of exposure.

H2O2 gas plasma vapo sterilization : (FUMIGATION)

 Hydrogen peroxide sterilization, also known as hydrogen peroxide gas sterilization, is
a low temperature sterilization process commonly used to sterilize heat-sensitive
devices. Once the sterilization cycle has completed, the vapor is vacuumed from the
chamber and converted to water and oxygen.
 It is most often used for plasma cleaning and sterilization. Hydrogen Peroxide in
liquid form is most known for its cleaning or bleaching properties. When an
aqueous solution of Hydrogen Peroxide is dosed into a plasma machine vacuum
chamber it evaporates and disperses.
 A hydrogen peroxide sterilization cycle typically requires less time than
alternative forms of sterilization, such as ethylene oxide sterilization. A hydrogen
peroxide sterilization process involves H2O2 vapor filling the sterilizer chamber,
contacting and sterilizing exposed device surfaces.
- 12 -
Previous years questions :

Aldehyde disinfectants :
(Formaldehyde, Paraformaldehyde, Glutaraldehyde) Formaldehyde – and its
polymerized solid paraformaldehyde have broad-spectrum biocidal activity and are both
effective for surface and space decontamination. As a liquid (5% concentration),
formaldehyde is an effective liquid decontaminant.

Antisepsis :
Antisepsis is the process of preventing the growth of infectious germs, like
bacteria, viruses and fungi. The substance used in antisepsis is called an antiseptic, which is
used to kill a broad range of infections.

Bio medical segregate according to 1998 bio medical rules :

1. Bio-medical waste shall not be mixed with other wastes.
2. Bio-medical waste shall be segregated into containers/bags at the points of
generation in accordance with Schedule II prior to its storage transportation, treatment and
disposal. Biological indicators :
Biological indicators (within a PCD) are often used for routine monitoring,
qualification and load monitoring of a steam sterilizer. Biological indicators are designed to
- 13 -
demonstrate whether the conditions during a steam (autoclave) cycle were adequate to
achieve a defined level of microbial inactivation.
examples :
anchor worm, salmon louse

Bowie dick test :

The Bowie-Dick Test is a standard operational test by which hospitals and
laboratories can demonstrate proper air removal from their pre-vacuum autoclave chamber. It
is primarily useful for testing pre-vacuum cycles that are sterilizing wrapped goods or packs.

Decontamination stages :
 Physical cleaning.
 Water purification.
 Ultrasonic cleaning.
 Disinfection.
 Antisepsis.
 Sterilization

Fumigation :
 Wear a gown, cap, mask and utility gloves
 Clean blood spills, remove waste, clean and disinfect items used in surgery
 Inspect all surfaces in the OT in detail for visible soiling/dust. Clean any soiling with
an HLD
 Wipe and clean all equipment completely i.e. wipe the entire OT table, OT lights,
trolleys, anaesthesia machine
 Lastly, clean and mop the floor twice (scrub by hand or a floor scrubber machine if
possible) with a high-level disinfectant beginning at the end farthest from the door and
moving towards it
 Cover all electronic equipment with plastic overs. The fogged liquid should not enter
them
 Turn off the ventilation system
 Fog the OT with "an aldehyde based product containing Glutaraldehyde and
chemically bound formaldehyde as principal disinfecting agents" e.g. Bacillocid*
until a fog is seen in the air. Stop the fogging and exit from the OT with the machine
 Thereafter, the OT should be closed
 The OT should not be entered after it has been closed down for the day (except for
emergency cases).

Principles of hand washing :

Hands of staff are the most common route by which microorganisms are transferred
between patients.Transient microorganisms acquired by touch are readily removed by soap
and water, and by alcohol-based hand rubs or gels.An effective technique for routine
handwashing involves three stages: preparation, washing and rinsing, and drying.
principles : (asepsis)
 Only sterile items are used within sterile fields. All articles used in anoperation have b
een sterilized previously.
- 14 -
  Persons who are sterile touch only sterile items/ areas; persons who are notsterile touc
h only unsterile items/ areas.
 If in doubt about sterility of anything, consider it unsterile.
 Nonsterile persons avoid reaching over a sterile field; sterile persons avoid
leaning over unsterile area.
 Tables are sterile only at table levels.
 Gowns are considered sterile only from waist to shoulder level in front, andthe sleev
 The edge of anything that encloses sterile contents is unsterile.
 Sterile persons keep well within sterile areas.
 Nonsterile persons keep away from sterile areas.
 Sterile field is created as close as possible to the time of use.
 Sterile areas are continuously kept in view.
 Destruction of integrity of microbial barriers results in contamination.Moisture can ca
use contamination.
 When microorganisms cannot be eliminated , they must be kept to an
irreducible minimum.

Incineration :
Incineration is a waste treatment process that involves the combustion of organic
substances contained in waste materials. Incineration and other high-temperature waste
- 15 -
treatment systems are described as "thermal treatment". Incineration of waste materials
converts the waste into ash, flue gas and heat.

Laminar flow :

In a laminar flow hood the air is passed

through a HEPA (High Efficiency
Particulates Air) filter which removes all
airborne contamination to maintain sterile
conditions......Now the sterile air flows into the
working (flasking) area where you can do all
your flasking work without risk of
contamination.

Ozone sterilization :
Ozone is the most powerful oxidative agent that occurs naturally. With its extra free
radical oxygen molecule, ozone is able to destroy germs, viruses, and microbes that may
cause surface or air contaminations.
advantages :
highly efficient at killing bacteria, viruses, and protozoa and is always generated on-site. It
does not require transportation or storage of dangerous materials. As it disinfects, it oxidises
inorganic and organic impurities such as iron and manganese.
disadvantages:
Low dosages may not effectively inactivate some viruses, spores, and cysts. * Ozonation is
more complex than other disinfection technologies. Ozone is very reactive and corrosive,
thus requiring corrosion-resistant material, such as stainless steel.

Pasteurisation :
Pasteurization or pasteurisation is a process in which water and certain packaged and
non-packaged foods (such as milk and fruit juice) are treated with mild heat, usually to less
than 100 °C (212 °F), to eliminate pathogens and extend shelf life.

Physical method of sterilization :

fumigation + h2o2 gas vap

Post exposure prophylaxis :

PEP (post-exposure prophylaxis) means taking antiretroviral medicines
(ART) after being potentially exposed to HIV to prevent becoming infected. PEP should be
- 16 -
used only in emergency situations and must be started within 72 hours after a recent possible
exposure to HIV.

Sunlight sterilization :
It is responsible for spontaneous sterilization in natural conditions. In tropical
countries, the sunlight is more effective in killing germs due to combination of
ultraviolet rays and heat. By killing bacteria suspended in water, sunlight provides
natural method of disinfection of water bodies such as tanks and lakes.

Hot air oven :

Hot air ovens are electrical devices which use dry heat to sterilize. They were
originally developed by Pasteur. Generally, they use a thermostat to control the temperature.
Their double walled insulation keeps the heat in and conserves energy, the inner layer being
a poor conductor and outer layer being metallic.
There is also an air filled space in between to aid insulation. An air circulating fan
helps in uniform distribution of the heat. These are fitted with the adjustable wire mesh
plated trays or aluminium trays and may have an on/off rocker switch, as well as indicators
and controls for temperature and holding time.
The capacities of these ovens vary. Power supply needs vary from country to
country, depending on the voltage and frequency (hertz) used. Temperature sensitive tapes or
biological indicators using bacterial spores can be used as controls, to test for the efficacy of
the device during use.

Advantages and disadvantages :

They do not require water and there is not much pressure build up within the
oven, unlike an autoclave, making them safer to work with. This also makes them more
suitable to be used in a laboratory environment. They are much smaller than autoclaves but
can still be as effective. They can be more rapid than an autoclave and higher temperatures
can be reached compared to other means.
As they use dry heat instead of moist heat, some organisms like prions, may not be
killed by them every time, based on the principle of thermal inactivation by oxidation.

Usage :
A complete cycle involves heating the oven to the required temperature,
maintaining that temperature for the proper time interval for that temperature, turning the
machine off and cooling the articles in the closed oven till they reach room temperature. The
standard settings for a hot air oven are:
 1.5 to 2 hours at 160 °C (320 °F)
 6 to 12 minutes at 190 °C (374 °F)
...plus the time required to preheat the chamber before beginning the
sterilization cycle. If the door is opened before time, heat escapes and the process becomes
incomplete. Thus the cycle must be properly repeated all over.

These are widely used to sterilize articles that can withstand high temperatures and not get
burnt, like glassware and powders. Linen gets burnt and surgical sharps lose their sharpness.

- 17 -
Nosocomial infection (hospital acquired infection ):
Types :
Hospital-acquired pneumonia
Ventilator-associated pneumonia
Urinary tract infection
Gastroenteritis
Puerperal fever
Central line-associated blood stream infection

Organisms :
Staphylococcus aureus
Methicillin resistant Staphylococcus aureus
Candida albicans
Pseudomonas aeruginosa
Acinetobacter baumannii'
Stenotrophomonas maltophilia
Clostridium difficile
Escherichia coli
Tuberculosis
Vancomycin-resistant Enterococcus
Legionnaires' disease

PREVENTION :
Sterilization :
Sterilization goes further than just sanitizing. It kills all microorganisms on equipment and
surfaces through exposure to chemicals, ionizing radiation, dry heat, or steam under pressure.

Isolation:
Isolation is the implementation of isolating precautions designed to prevent transmission of
microorganisms by common routes in hospitals. Because agent and host factors are more
difficult to control, interruption of transfer of microorganisms is directed primarily at
transmission for example isolation of infectious cases in special hospitals and isolation of
patient with infected wounds in special rooms also isolation of joint transplantation patients
on specific rooms.

Hand washing :
 Hand washing frequently is called the single most important measure to reduce the
risks of transmitting skin microorganisms from one person to another or from one
site to another on the same patient.
 Washing hands as promptly and thoroughly as possible between patient contacts and
after contact with blood, body fluids, secretions, excretions, and equipment or articles
contaminated by them is an important component of infection control and isolation
precautions.
 The spread of nosocomial infections, among immune compromised patients is
connected with health care workers' hand contamination in almost 40% of cases,
and is a challenging problem in the modern hospitals.
- 18 -
  Two categories of micro-organisms can be present on health care workers' hands:
transient flora and resident flora.
 The first is represented by the micro-organisms taken by workers from the
environment, and the bacteria in it are capable of surviving on the human skin
and sometimes to grow. The second group is represented by the permanent micro-
organisms living on the skin surface .
 Improving patient hand washing has also been shown to reduce the rate of
nosocomial infection. Patients who are bed-bound often do not have as much access
to clean their hands at mealtimes or after touching surfaces or handling waste such as
tissues.
 All visitors must follow the same procedures as hospital staff to adequately control
the spread of infections. Moreover, multidrug-resistant infections can leave the
hospital and become part of the community flora if steps are not taken to stop this
transmission.

Gloves :
In addition to hand washing, gloves play an important role in reducing the risks of
transmission of microorganisms.
Gloves are worn for three important reasons in hospitals.
 First, they are worn to provide a protective barrier for personnel,
preventing large scale contamination of the hands when touching blood,
body fluids, secretions, excretions, mucous membranes, and non-intact
skin.
 Second, gloves are worn to reduce the likelihood that microorganisms present
on the hands of personnel will be transmitted to patients during invasive or other
patient-care procedures that involve touching a patient's mucous membranes and
nonintact skin.
 Third, they are worn to reduce the likelihood that the hands of personnel
contaminated with micro-organisms from a patient or a fomite can transmit those
micro-organisms to another patient. In this situation, gloves must be changed
between patient contacts, and hands should be washed after gloves are removed.

Surface sanitation :
Sanitizing surfaces is part of nosocomial infection in health care environments.
Modern sanitizing methods such as Non-flammable Alcohol Vapor in Carbon Dioxide
systemshave been effective against gastroenteritis, MRSA, and influenza agents. Use
of hydrogen peroxide vapor has been clinically proven to reduce infection rates and risk of
acquisition. Hydrogen peroxide is effective against endospore-forming bacteria, such
as Clostridium difficile, where alcohol has been shown to be ineffective.

Antimicrobial surfaces :
Micro-organisms are known to survive on inanimate ‘touch’ surfaces for extended
periods of time. This can be especially troublesome in hospital environments where patients
with immune deficiencies are at enhanced risk for contracting nosocomial infections.

Touch surfaces commonly found in hospital rooms, such as bed rails, call buttons, touch
- 19 -
plates, chairs, door handles, light switches, grab rails, intravenous poles, dispensers (alcohol
gel, paper towel, soap), dressing trolleys, and counter and table tops are known to be

- 20 -
contaminated with Staphylococcus, MRSA (one of the most virulent strains of antibiotic-
resistant bacteria) and vancomycin-resistant Enterococcus (VRE). Objects in closest
proximity to patients have the highest levels of MRSA and VRE. This is why touch surfaces
in hospital rooms can serve as sources, or reservoirs, for the spread of bacteria from the
hands of healthcare workers and visitors to patients.

Types of sterilization :
5 Laboratory Sterilisation Methods
 WET HEAT (Autoclaving) The method of choice for sterilisation in most labs is
autoclaving; using pressurised steam to heat the material to be sterilised.

 DRY HEAT (Flaming, baking) Dry heating has one crucial difference from
autoclaving.

 FILTRATION.

 SOLVENTS.

 RADIATION.

THE END

PREPARED BY ,
P.M. AMJATH ALTHAF (B.Sc operation theatre and anaesthesia technology )
Contact : 88708 25976

- 21 -