Control of Microorganism

Md. Ariful Islam Juel

Lecturer, LE, KUET
11.1 Controlling Microorganisms
 General Considerations in Microbial Control

 Sterilization: the complete removal or destruction

of all microbial life used on inanimate objects
 Disinfection: The destruction or removal of
vegetative pathogens but not bacterial endospores.
Usually used only on inanimate objects.
 Antisepsis: Chemicals added to body surface to
destroy or inhibit vegetative pathogens.
 Decontamination: the mechanical removal of most
microbes from an animate or inanimate surface
Microbial Control Methods
Relative Resistance of Microbial Forms

 Primary targets of microbial control: microorganisms

that can cause infection or spoilage that are constantly
present in the external environment
 Highest resistance
 Bacterial endospores
 Moderate resistance
 Protozoan cysts, fungal sexual spores, naked viruses,
resistant vegetative bacteria
 Least resistance
 Most bacterial vegetative cells, fungal spores, enveloped
viruses, yeasts.
Comparison of Resistance
Terminology and Methods of Microbial Control

 Sterilization
 Removes all viable microorganisms including
viruses and bacterial endospores
 Material is said to be sterile

 Usually reserved for inanimate objects

 Mostly performed with heat

 Sometimes chemicals called sterilants are used

Disinfection
 The use of a physical process or chemical agent
(disinfectant) to destroy vegetative pathogens
 Does not destroy bacterial endospores

 Usually used only on inanimate objects

 Also removes toxins

 5% bleach solution, boiling water, iodine solutions

Antisepsis

 Antiseptics: applied directly to exposed body

surfaces to destroy or inhibit vegetative pathogens
 Sepsis: the growth of microorganisms in the blood
and other tissues
 Asepsis: any practice that prevents the entry of
infectious agents into sterile tissues
The Agents Versus the Processes

 –cide: to kill
 Bactericide: chemical that destroys bacteria (not endospores)
 Fungicide: a chemical that can kill fungal spores, hyphae, and
yeasts
 Virucide: a chemical that inactivates viruses
 Sporicide: can destroy bacterial endospores
 Germicide and microbicide: chemical agents that kill
microorganisms
 Stasis and static: to stand still, prevent multiplication
 Bacteristatic, Fungistatic
 Microbiostatic: materials used to control microorganisms in
the body, for example
Decontamination
 Used when actual sterilization isn’t needed but need to
decrease the risk of infection or spoilage (ex. food
industry)
 Sanitization: any cleansing technique that
mechanically removes microorganisms to reduce
contamination to safe levels
 Sanitizer: compound such as soap or detergent that
sanitizes
 Sanitary: may not be free from microbes but are safe
for normal use
 Degermation: reduces the numbers of microbes on
the human skin (ex. alcohol wipes)
How Antimicrobial Agents Work: Their Modes
of Action

 The Cell Wall

 The Cell Membrane

 Protein and Nucleic Acid Synthesis

 Protein Function
Antimicrobial Effects on Cell Well

 Block its synthesis

 Digest it

 Break down its surface

 The cell becomes fragile and is lysed easily.

Cell Membrane

 All microorganisms
have cell membrane
 Many viruses as well

 Disrupt cell membrane=

cell loss of selective
permeability
 Detergents (surfactants)
disrupt membrane
Protein and Nucleic Acid Synthesis
 Any level can be affected: Replication, Transcription,
and/or Translation
 Some agents bind to ribosomes to stop translation

 Some agents bind irreversibly to DNA preventing

transcription and translation
 Mutagenic agents
Protein Function

 Proteins must be in
native state
 Disrupt the native state
(denature)
 Break the bonds of
secondary or tertiary
structures
 Coagulation: heat,
alcohol, acids, phenolics
11.2 Methods of Physical Control

 Heat as an Agent of Microbial Control

 Generally, elevated temperatures are microbicidal
and lower temperatures are microbiostatic
 Can use moist heat or dry heat

 Moist heat is more effective

Mode of Action
 Moist coagulation and denaturation
 Dry remove water from organisms, incineration
Moist vs. Dry Heat
Common Methods of Moist Heat Control

 Steam under pressure

 Nonpressurized steam

 Pasteurization

 Boiling water
Steam Under Pressure

 Pressure raises the

temperature of steam
 Autoclave is used

 Most efficient pressure-

temperature combination for
sterilization: 15 psi which
yields 121°C
 Kills all endospores

 Home pressure cookers- same

principle
Nonpressurized Steam
 Intermittent sterilization or tyndallization

 This process involves steaming media for 30 minutes

each day for three successive days and incubating them
in between.
 The principle is that the spores which may escape first
steaming will germinate at incubator temperature and
will be killed in the next steaming.
 A single exposure is not sufficient to kill all spores

 Used some for canned foods and certain media

preparations
Pasteurization
 Used to disinfect beverages

 Heat is applied to liquids to kill potential agents of infection

and spoilage, while retaining the liquid’s flavor and food
value
 Special heat exchangers
 Flash method: expose to 71.6°C for 15 seconds

 Batch method: expose to 63°C to 66°C for 30 minutes

 Does not kill endospores or thermoduric microbes

 Pasteurized milk is NOT sterile

Boiling Water
 For disinfection and not sterilization

 Expose materials to boiling water for 30 minutes

Dry Heat: Hot Air and Incineration
 Incineration
 Ignites and reduces
microbes to ashes and gas
 Common practice in
microbiology lab-
incineration on
inoculating loops and
needles using a Bunsen
burner
Dry Oven
 Usually an electric oven

 Coils radiate heat within an enclosed compartment

 Exposure to 150°C to 180°C for 2 to 4 hours

 Used for heat-resistant items that do not sterilize well

with moist heat
 Glassware, metallic instruments
Modes of Action of Ionizing Versus Nonionizing
Radiation

 Ionizing: ejects
electron, causing ions
to form
 Nonionizing: excites
atoms but does not
ionize them
 Ionizing Radiation

 Gamma rays, most penetrating

 X rays, intermediate

 cathode rays, least penetrating

Applications of Ionizing Radiation

 Food products

 Medical products

 Main advantages:
 Speed

 Penetrating power

 No heat
Nonionizing Radiation: Ultraviolet Rays

 Wavelength approximately 100 nm to 400 nm

 Germicidal lamp: 254 nm

 Not as penetrating as ionizing radiation

 Powerful tool for destroying fungal cells and

spores, bacterial vegetative cells, protozoa, and
viruses
Applications of Ultraviolet Radiation

 Usually disinfection
rather than sterilization
 Hospital rooms,
operating rooms,
schools, food prep
areas, dental offices
 Treat drinking water or
purify liquids
 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

Filtration
Liquid

 Pass liquid or gas through

a filter with sufficiently Filter Pore

small pore size

 HEPA – high-efficiency
particulate air Filter

 Advantages: No thermal Sterilized Vacuum

fluid

damage
Pump suction
(a)

 Disadvantages: viruses
not eliminated and must
be either liquid or gas
(b)
b: © Fred Hossler/Visuals Unlimited
11.3 Chemical Agents in Microbial Control

 Approximately 10,000 different antimicrobial

chemical agents are manufactured
 Approximately 1,000 used routinely in health care
and the home
 Occur in liquid, gaseous, or solid state

 Tinctures: solutions dissolved in pure alcohol or

water-alcohol mixtures
Concentration and Time Needed
Phenol and its Derivatives

 Cresols, detols, lysols are common example.

 A 2% solution of lysol is generally used as a

germicidal lotion for contaminated material after use
in bacteriological laboratory.
 0.5% phenol is used for vaccine preservation.
 Alcohols

 Ethyl alcohol is effective(bactericidal) against

vegetative bacteria but not against spores.
Methylated spirit is commonly used. 70% alcohol in
water is bactericidal.
 Formaldehyde, a derivative of methyl alcohol can
be used in gaseous form in disinfecting rooms &
rubber goods. Formaline is a powerfull disinfecting
agents.
Alcohols as Antimicrobial Agents

 Only ethyl and isopropyl alcohols are suitable for

microbial control
 Mechanism of action depends in part upon its
concentration
 Does not destroy bacterial spores at room
temperature but can destroy resistant vegetative
forms
 More effective in inactivating enveloped viruses
than nonenveloped viruses
Hydrogen Peroxide and Related Germicides
 Germicidal effects are due
to the direct and indirect
actions of oxygen
 Oxygen forms hydroxyl free
radicals which are highly
toxic and reactive to cells
 Can be harmful to tissue

 Bactericidal, virucidal, and

fungicidal
 In higher concentrations is
sporicidal
Chemicals with Surface Action: Detergents

 Act as surfactants
 Cationic detergents are more
effective because the
positively charged end binds
well with the predominantly
negatively charged bacterial
surface proteins
 Soaps are weak
microbicides but gain
germicidal value when
mixed with agents such as
chlorhexidine or iodine
Heavy Metal Compounds

 Hg, Ag, Au, Cu, As, and Zn have been used

 Oligodynamic action: having antimicrobial effects in
exceedingly small amounts
 Bind onto functional groups of proteins and inactivating
them
 Drawbacks to using metals in microbial control:
 Can be very toxic to humans
 Often cause allergic reactions
 Large quantities of biological fluids and wastes neutralize
their actions
 Microbes can develop resistance to them
Germicidal Effect of Heavy Metals
Aldehydes as Germicides

 –CHO functional group

on the terminal carbon
 Glutaraldehyde and
formaldehyde
(formalin- aqueous
solution)- most often
used in microbial
control
Gaseous Sterilants and Disinfectants

 Ethylene oxide (ETO)

 Propylene oxide

 Chlorine dioxide

 Block DNA replication, protein function

Dyes as Antimicrobial Agents
 Primary source of certain drugs used in
chemotherapy
 Aniline dyes (crystal violet and malachite green) are
very active against gram-positive species of bacteria
and various fungi
 Yellow acridine dyes (acriflavine and proflavine)
sometimes used for antisepsis and wound treatment
 Limited applications because they stain and have a
narrow spectrum of activity
Acids and Alkalis
 Very low or high pH can destroy or inhibit microbial
cells
 Limited in applications due to their corrosive,
caustic, and hazardous nature
Main Ingredients Antimicrobial Products