Oleh:

Tim Dosen Mikrobiologi Farmasi - ISTN

 Berikut adalah beberapa istilah dalam
pengendalian mikroorganisme

A. BIOCIDE
 A general term describing a chemical agent,

usually broad-spectrum, that inactivates

microorganisms.
 Biocides can be antiseptics, disinfectants, or

preservatives.
The activity of biocides against microorganisms
depends on:
1) the external physical environment;
2) The nature, structure, composition, and
condition of the microorganism itself
3) the ability of the microorganism to degrade or
inactivate the biocide.
B. BACTERIOSTATIC
 A specific term referring to the property by which

a biocide is able to inhibit bacterial multiplication;

multiplication resumes upon removal of the agent.
 The terms “fungistatic” and “sporostatic” refer to

biocides that inhibit the growth of fungi and

spores, respectively.
C. BACTERICIDAL
 A specific term referring to the property by which a

biocide is able to kill bacteria.

 Bactericidal action differs from bacteriostasis only in

being irreversible; ie, the “killed” organism can no

longer reproduce, even after being removed from
contact with the agent.
 In some cases, the agent causes lysis (dissolution) of

the cells; in other cases, the cells remain intact and

may even continue to be metabolically active.
 (The terms “fungicidal,” “sporicidal,” and “virucidal”

refer to the property whereby biocides are able to kill

fungi, spores, and viruses, respectively).
D. STERILIZATION
• A physical or chemical process that completely
destroys or removes all microbial life, including
spores.

E. DISINFECTION
• is the destruction of pathogenic microorganisms by
processes that fail to meet the criteria for
sterilization.
• Disinfection uses chemical agents to kill pathogens
with varying efficiency known as DISINFECTANTS.
• Disinfectants are not necessarily sporicidal, but are
sporostatic, inhibiting germination or outgrowth.
F. SEPTIC
 Characterized by the presence of pathogenic

microbes in living tissue.

G. ANTISEPTIC
 are disinfectant agents that can be used on body

surfaces such as the skin or vaginal tract to

reduce the numbers of normal flora and
pathogenic contaminants.
 They have lower toxicity than disinfectants used

environmentally but are usually less active in

killing vegetative organisms.
H. ASEPTIC
 Characterized by the absence of pathogenic

microbes.
 describes processes designed to prevent

microorganisms from reaching a protected

environment.

I. SANITIZATION
 is a less precise term with a meaning somewhere

between disinfection and cleanliness. It is used

primarily in housekeeping and food preparation
contexts.
J. PRESERVATION
 The prevention of multiplication of

microorganisms in formulated products,

including pharmaceuticals and foods.

K. ANTIBIOTICS
 Naturally occurring or synthetic organic

compounds which inhibit or destroy

selective bacteria
A. DAMAGE TO DNA
 A number of physical and chemical agents act by
damaging DNA; these include ionizing radiations,
ultraviolet light, and DNA-reactive chemicals.
 Among the last category are alkylating agents and
other compounds that react covalently with purine
and pyrimidine bases to form DNA adducts or
interstrand cross-links.
B. PROTEIN DENATURATION
 a number of noncovalent linkages such as ionic,
hydrophobic, and hydrogen bonds. This state is
called the tertiary structure of the protein;
 it is readily disrupted by a number of physical or
chemical agents, Denaturation known as the
disruption of the tertiary structure of a protein.
 Proteins exist in a folded, three-dimensional state
determined by intramolecular covalent disulfide
linkages and causing the protein to become
nonfunctional.
C. DISRUPTION OF CELL MEMBRANE OR WALL
 Substances that concentrate at the cell surface
may alter the physical and chemical properties
of the membrane, preventing its normal
functions and therefore killing or inhibiting the
cell.
 The cell wall acts as a corseting structure,
protecting the cell against osmotic lysis. Thus,
agents that destroy the wall (eg, lysozyme) or
prevent its normal synthesis (eg, penicillin) may
bring about lysis of the cell.
D. REMOVAL OF FREE SULFHYDRYL GROUPS
• Enzyme proteins containing cysteine have side chains
terminating in sulfhydryl groups.
• In addition to these, coenzymes such as coenzyme A and
dihydrolipoate contain free sulfhydryl groups.
• Such enzymes and coenzymes cannot function unless the
sulfhydryl groups remain free and reduced.
• Oxidizing agents thus interfere with metabolism by forming
disulfide linkages between neighboring sulfhydryl groups:
Many metals such as mercuric ion likewise interfere by
combining with sulfhydryls: There are many sulfhydryl
enzymes in the cell; therefore, oxidizing agents and heavy
metals do widespread damage.
E. CHEMICAL ANTAGONISM
 The interference by a chemical agent with the normal
reaction between a specific enzyme and its substrate is
known as “chemical antagonism.”
 The antagonist acts by combining with some part of the
holoenzyme (either the protein apoenzyme, the mineral
activator, or the coenzyme), thereby preventing attachment
of the normal substrate.
 for example, carbon monoxide and cyanide combine with
the iron atom in heme-containing enzymes and prevent
their function in respiration.
 Sterilisasi merupakan proses penghancuran atau
penghilangan mikroorganisme, termasuk bakteri
dan sporanya.
 Sterilisasi medium kultur dan peralatan kerja
dalam kerja mikrobiologi sangatlah penting untuk
isolasi dan perawatan mikroorganisme.
 Dalam proses pengobatan dan operasi, sterilisasi
alat, obat-obatan dan barang lainnya penting
untuk pencegahan infeksi.
Sterilisasi dapat dilakukan secara
I. Fisika
A. Heat/Pemanasan
1) Dry heat = pemanasan kering
2) Moist heat = pemanasan basah
B. Radiations /Radiasi
1) Ultraviolet radiations = UV
2) Ionizing radiations
C. Filtration/Penyaringan
II. Kimia
A. Ethylene oxide
B. Disinfectants
 Sterilisasi panas merupakan metode yang paling
dipercaya dan banyak digunakan untuk bahan
yang tahan panas.
 Metode sterilisasi panas dengan penggunaan
uap air disebut sterilisasi panas lembap/basah;
sedangkan tanpa kelembapan (tanpa
penggunaan uap air) disebut sterilisasi panas
kering.
 Panas kering: suhu 160 C – 180 C
 Panas basah: suhu 115 C – 134 C
• Proses sterilisasi panas ini terdiri atas tiga tahap,
yaitu:
1. Tahap pemanasan (heating stage): peningkatan
suhu bahan yang disterilisasi
2. Tahap sterilisasi (holding stage): waktu yang
diperlukan untuk proses sterilisasi
3. Tahap pendinginan (cooling stage): waktu yang
diperlukan untuk penurunan suhu bahan yang
disterilisasi
FACTORS INFLUENCING STERILIZATION BY HEAT
1.The temperature and time: they are inversely
related, shorter time is sufficient at high
temperatures.
2.Number of microorganisms and spores: The
number of survivors diminished exponentially with
the duration of heating
3.Depends on the species, strains and spore
forming ability of the microbes.
4. Thermal death point is the lowest temperature
to give complete killing in aqueous suspension
within 10 minutes.
5. Depends on the nature of material: a high
content of organic substances generally tends to
protect spores and vegetative organisms against
heat.
6. Presence of organic or inorganic disinfectants
facilitates killing by heat
7. pH also plays an important role in the killing of
microorganisms
Mechanism of killing by dry heat
Mematikan organisme dengan cara

mengoksidasi komponen sel ataupun

mendenaturasi enzim.
Dry heat is employed for glassware; syringes,

metal instruments and paper wrapped goods,

which are not spoiled by high temperatures.
It is also used for anhydrous fats, oils and powders

that are impermeable to moisture.

Tidak dapat digunakan untuk bahan-bahan yang

terbuat dari karet atau plastik.

 Metode ini tidak memerlukan air sehingga tidak
ada uap air yang membasahi alat atau bahan yang
disterilkan
 Ada dua metode sterilisasi panas kering, yaitu
dengan insinerasi (pembakaran menggunakan api
bunsen, suhu 350 C), dan panas Oven (suhu 160-
170 C).
 RED HEAT
 Inoculating wires, points of forceps and searing spatulas
are sterilized by holding them in the flame of Bunsen
burner until they are seen to be red-hot.

 FLAMING
 This method is used for sterilizing scalpel, mouth of
culture tubes, glass slides etc.
 It involves passing of an article through Bunsen flame
without allowing it to become red-hot.
STERILIZATION BY DRY HEAT: OVEN
 This is the main means of sterilization by
dry heat.
 Exposure at a temperature of 160 °C for 2-
3 hours is generally employed.
 Low penetrate
• Moist heat kills the organisms by coagulating and
denaturing their enzymes and structural protein.
• Moist heat is used for the sterilization of culture
media, and all other materials through which
steam can penetrate
• Moist heat is more effective than dry heat
Moist heat can be employed at:
 Temperature below 100 °C:
 Industri makanan: 60-800 C
 Susu: 630 C selama 30 menit atau 720 C, 15 menit
 Plasma manusia: pasteurisasi 600 C, selama 10 menit
 Temperature of 100 °C
 Peralatan dan cairan: selama 5-10 menit
 Perebusan menggunakan air mendidh 1000 C selama 10
menit efektif untuk sel-sel vegetatif dan spora eukariot,
tetapi tidak efektif untuk endospora bakteri.
 Temperature above 100 °C
 autoklaf
o In Pasteurization of milk the temperature employed is either
63 °C for 30 minutes or 72 °C for 20 seconds.
o All nonspore-forming pathogens in milk like Salmonellae,
M.tuberculosis are killed.
 High temperatures
can cause damage
to the taste,
texture and
nutritional value of
many food
substances and in
such instances, it is
sufficient to
destroy vegetative
cells by a process
of pasteurisation.
[Sumber: Barraquio, 2014]
 It is the method most widely used for sterilization of
culture media and surgical supplies
 Prinsip: terjadi koagulasi yang lebih cepat dalam
keadaan basah dibandingkan kering.
 Dapat membunuh mikroorganisme dengan cara
mendenaturasi enzim atau mengkoagulasi protein
pada enzim dan membran sel mikroorganisme.
 Normally autoclaving: 115 °C for 15 minutes
 An effect similar to that achieved by autoclaving
can be obtained by a method called intermittent
steaming or tyndallisation.
 This is used for those substances or materials
that might be damaged by the high
temperatures used in autoclaving.
 The material is heated to between 90 and 1000 C
for about 30 minutes on each of three successive
days, and left at 370 C in the intervening periods.
 Metode sterilisasi dengan
menghilangkan kandungan
air
 A desiccator is used for the
cooling of heated objects
and for the storage of dry
objects that must not be
exposed to the moisture
normally present in the
atmosphere.
Ultraviolet Light and Ionizing
Radiation
 Ultraviolet (UV) light in the wavelength range
240 to 280 nm is absorbed by nucleic acids and
causes genetic damage, including the formation
of the thymine dimers
 UV radiation has very poor penetrating powers; a
thin layer of glass, paper or fabric is able to
impede the passage of the rays.
 UV lamps are commonly found in food
preparation areas, operating theatres and
specialist areas such as tissue culture facilities,
where it is important to prevent
contamination.
 Because they are also harmful to humans
(particularly the skin and eyes), UV lamps can
only be operated in such areas when people
are not present.
Ionizing radiation carries far greater energy than
UV light. It, too, causes direct damage to DNA
and produces toxic free radicals and hydrogen
peroxide from water within the microbial cells.
Ionisingradiations have a shorter wavelength and
much higher energy, giving them greater
penetrating powers.
Use for surgical supplies, food
 Cathode rays and gamma rays from cobalt-
60 are widely used in industrial processes,
including the sterilization of many disposable
surgical supplies such as gloves, plastic
syringes, specimen containers, some
foodstuffs, and the like, because they can be
packaged before exposure to the penetrating
radiation.
 Gamma radiation has been approved for use in
over 40 countries for the preservation of food,
which it does not only by killing pathogens and
spoilage organisms but also by inhibiting
processes that lead to sprouting and ripening.
 Irradiation is not suitable for some foodstuffs,
such as those with a high fat content, where
unpleasant tastes and odours result.
 Ionising radiations have the great advantage
over other methods of sterilisation that they
can penetrate packaging.
 Liquids that would be damaged by heat such as
serum and antibiotic solutions can be sterilized
by filtration
 When fluids are passed through bacteria
stopping filters, they are made free from
bacteria.
 It is useful for making preparations of soluble
products of bacterial growth such as toxins
 Efficient filters should be able to retain Serratia
marcescens
 Filtersused to be made from materials such as
asbestos and sintered glass, but have been largely
replaced by membrane filters, commonly made of
nitrocellulose or polycarbonate
A pore size of 0.22μm is commonly used; this will
remove bacteria plus, of course, anything bigger, such
as yeasts; however, Mycoplasma and viruses are able
to pass through pores of this size.
 With a pore size 10 times smaller than this, only the
smallest of viruses can pass through, so it is
important that an appropriate pore size is chosen for
any given task.
 There are different kinds of filters
1. Earthenware candles - called Berkfield &
Chamberland filters
2. Asbestos and asbestos-paper discs filters -
called Seitz filters
3. Sintered glass filters
4. Cellulose membrane filters
5. Fibre glass filters.
 SEITZ FILTER

SINTERED GLASS FILTER

ASBESTOS DISCS
 Membrane filters
 Made up of nitro-cellulose membranes
 Made with different grades of porosity by
adjusting the concentration of constituents
• The use of the gas ethylene oxide, however, is effective
against bacteria, their spores and viruses.
• It is used for sterilising large items of medical
equipment, and materials such as plastics that would be
damaged by heat treatment.
• Ethylene oxide is particularly effective in sterilising
items such as dressings and mattresses, due to its great
powers of penetration.
• In the food industry, it is used as an antifungal fumigant,
for the treatment of dried fruit, nuts and spices.
 The materials to be treated are placed in a special
chamber which is sealed and filled with the gas in a
humid atmosphere at 40–50 ◦C for several hours.
 Ethylene oxide is highly explosive, so it must be
used with great caution; its use is rendered safer by
administering it in admixture (10 per cent) with a
non-flammable gas such as carbon dioxide.
 It is also highly toxic, so all items must be
thoroughly flushed with sterile air following
treatment to remove any trace of it.
 Ethylene oxide is an alkylating agent; it denatures
proteins by replacing labile hydrogens such as those
on sulphydryl groups with a hydroxyl ethyl radical
 Disinfection
is the elimination or inhibition of
pathogenic microorganisms in or on an object so that
they no longer pose a threat.
A disinfectant is a chemical agent used to disinfect
inanimate objects suchbas work surfaces and floors.
 Disinfectantsare incapable of killing spores within a
reasonable time period, and are generally effective
gainst a narrower range of organisms than physical
means.
 Decontamination is a term sometimes used
interchangeably with disinfection, but its scope is
wider, encompassing the removal or inactivation
of microbial products such as toxins as well as the
organisms themselves.
 The lethal action of disinfectants is mainly due to
their ability to react with microbial proteins, and
therefore enzymes.
 Ethanol
and isopropanol are most commonly used at a
concentration of 70 per cent.
 Aswell as denaturing proteins, alcohols may act by
dissolving lipids, and thus have a disruptive effect on
membranes, and on the envelope of certain viruses.
 Bothbacteria and fungi are killed by alcohol
treatment, but spores are often resistant because of
problems in rehydrating them; there are records of
anthrax spores surviving in ethanol for 20 years!
 The
use of alcohols is further limited to those
materials that can withstand their solvent action.
 Chlorine is an effective disinfectant as a free gas,
and as a component of chlorine releasing
compounds such as hypochlorite and chloramines.
 Chlorine gas, in compressed form, is used in the
disinfection of municipal water supplies,
swimming pools and the dairy industry.
 Sodium hypochlorite (household bleach) oxidises
sulphydryl (−SH) and disulphide (S−S) bonds in
proteins.
 Iodine acts by combining with the tyrosine
residues on proteins; its effect is enhanced by
being dissolved in ethanol (1 per cent I2 in 70 per
cent ethanol) as tincture of iodine, an effective
skin disinfectant.
 Its use is being superseded by iodophores
(Betadine, Isodine), in which iodine is combined
with an organic molecule, usually a detergent, to
combat bacteria, viruses and fungi, but not
spores.
 Since it is highly toxic, phenol’s use in the
disinfection of wounds has long since been
discontinued, but derivatives such as cresols and
xylenols continue to be used as disinfectants and
antiseptics.
 These are both less toxic to humans and more
effective against bacteria than the parent
compound.
 Phenolics act by combining with and denaturing
proteins, as well as disrupting cell membranes.
 Their advantages include the retention of activity
in the presence of organic substances and
detergents, and their ability to remain active for
some time after application; hence their effect
increases with repeated use.
 Familiar disinfectants such as Dettol, Lysol and
chlorhexidine (Hibitane, Hibiscrub) are all phenol
derivatives.
 A surfactant reduces the tension between two
molecules at an interface.
 Surface active agents or surfactants, such as soaps
and detergents, have the ability to orientate
themselves between two interfaces to bring them
into closer contact
 The value of soap has less to do with its
disinfectant properties than with ability to
facilitate the mechanical removal of dirt and
microorganisms.
 It does this by emulsifying oil secretions, allowing
the debris to be rinsed away.
 Detergents may be anionic (negatively charged),
cationic (positively charged) or nonionic.
 Cationic detergents such as quaternary
ammonium compounds act by combining with
phospholipids to disrupt cell membranes and
affect cellular permeability.
 Mikroorganisme memiliki sensitivitas berbeda-
beda terhadap metode sterilisasi tertentu.
 Endospora bakteri resisten terhadap panas,
iradiasi dan detergen.
 Mycoplasma dan virus tidak dapat dihilangkan
dengan filter steril (0,2 μm)