Microbicidal - Agents that used to kill

microorganisms
Control of Microbial Growth
 Germicide
Module 5
 Sporicide
 Fungicide
Terminology of Microbial Control  Virucide

Microbistatic - Agents used to inhibit growth

Commercial and multiplication or reproduction of
Sterilization Disinfection
Sterilization microorganism

Sepsis – Decay or putrid. Bacterial

Antisepsis Degermining Sanitation
Microbicidal Microbistatic Sepsis
Sterilization - The removal or destruction of all
forms of microbial life including endospores.
 Heating is the most common method of
killing microbes
 Membrane Filtration is the removal of
microbes from gases or liquids
 Steam under pressure
 Sterilizing gas like ethylene oxide
Commercial Sterilization – Sufficient heat
treatment to kill endospores of Clostridium
botulinum in canned food.
 More resistant endospores of thermophilic
bacteria may survive but they will not
germinate and grow under normal storage
condition
Disinfection – Destructive of vegetative
pathogens on inert substance or surface.
 Chemicals – “DISINFECTANT”
 IV Radiation
 Boiling water or steam
 Methods may either physical or chemical
Antisepsis - Destructive of vegetative
pathogens on living tissue
contamination.
 Aseptic - Area or object is free of
pathogens
 Asepsis - Absence of significant
contamination
 Aseptic Technique - Important in surgery
to minimize contamination from the
instruments, operating personnel, and
the patient
ACTIONS OF MICROBIAL CONTROL AGENTS
Alteration of Membrane Permeability
 PLASMA MEMBRANE
 Damage to the lipids or proteins
by antimicrobial agents
 Effect: Cellular contents to leak
into the cellular medium.
Interferes with the growth of the
cell.
Damage of Proteins and Nucleic Acids
 PROTEINS
 Enzymes
 Susceptible to breakage by heat or
certain chemicals (Hydrogen Bonds
and Covalent Bonds)
 NUCLEIC ACIDS
 DNA and RNA
 Damage caused by heat, chemicals
and radiation is lethal to the cell
(Can no longer replicate).

 Chemical – “ANTISEPTIC” MICROBIAL CONTROL

 Treatment is always by chemical
antimicrobials Physical Chemical
Methods Methods
Degermining - Mechanical removal rather than  Heat  Phenols and
killing of most microbes in the limited area  Filtration Phenolics
 Low Temperature  Biguanides
 Removal of microbes from the skin around (Cold)  Halogens
an injection site by an alcohol-soaked  High Pressure  Alcohol
swab  Desiccation  Heavy metals and
 Osmotic Pressure their compounds
Sanitation - Treatment intended to lower  Radiation  Surface active
microbial counts on eating and drinking agents
utensils to safe public health levels.  Organic Acids
 Aldehydes
 High temperature washing
 Gaseous Sterilants
 Dipping into chemical disinfectant
 Peroxygens
 PHYSICAL METHODS
HEAT
 The most practical, efficient and inexpensive
method of controlling microbial growth
 Factors affecting heat efficiency:
Temperature and Time
 THERMAL DEATH POINT (TDP) - Lowest
temperature at which all the
microorganisms in a particular liquid
suspension will be killed in 10 minutes
 THERMAL DEATH TIME (TDT) - Minimal length
of time for all bacteria in a particular liquid
culture to be killed at a given temperature
 Types: Moist Heat, Dry Heat Sterilization,
Pasteurization
MOIST HEAT– MOA: Protein denaturation
(coagulation)
 Boiling – Kills vegetative bacterial and
fungal pathogens and almost all
viruses within 10 minutes; less effective
on endospores. Applications: dishes,
basins, various equipment
 Autoclave - Very effective method of
sterilization: 121ºC at 15psi; all
vegetative cells and their endospores
are killed in about 15 min.
Applications: microbial media,
utensils, and others that can
withstand temperature and pressure
DRY HEAT
 Direct Flaming – MOA: burning
contaminants to ashes. Very effective
method of sterilization. Uses:
inoculating loop
 Incineration – MOA: burning to ashes.
Very effective method of sterilization.
Uses: paper cups, contaminated
dressings, animal carcasses and
wipes.
 Hot-air Sterilization – MOA: Oxidation.
Very effective method of sterilization
but requires temp of 170ºC for 2
hours. Uses: empty glassware,
instruments, needles and glass
syringes
PASTEURIZATION - MOA: Protein
denaturation. Temperature is 72ºC for
about 15 seconds that kills pathogens
and most nonpathogens. Uses: milk,
cream, beer, wine
FILTRATION - MOA: Separation of bacteria from
suspending liquid. Removes microbes by
passage of a liquid or gas through a screen-like
material. Uses: sterilizing liquids (enzymes,
vaccines) that are destroyed by heat.
COLD – MOA: decreased chemical reactions
and possible changes in proteins. Uses: food,
drug and culture preservation
 Refrigeration – Has bacteriostatic effect
 Deep-freezing - Effective method for
preserving microbial cultures in which
cultures are quick-frozen between -50ºC
and -95ºC
 Lyophilization - Most effective method
for long term preservation of microbial
cultures; water removed by high
vacuum at low temperature
HIGH PRESSURE - MOA: alteration of molecular
structure of proteins and carbohydrates.
Preservation of colors, flavors, nutrient values.
Uses: fruit juices
DESSICATION - MOA: disruption of metabolism.
Involves removing of water from microbes.
Bacteriostatic. Uses: food preservation
OSMOTIC PRESSURE - MOA: plasmolysis. Results
in loss of water from microbial cells. Uses: food
preservation
RADIATION
 Ionizing - MOA: destruction of DNA. Not
widespread in routine sterilization. Uses:
sterilizing pharmaceuticals and medical
and dental supplies.
 Non-ionizing - MOA: damage to DNA.
Radiation not very penetrating. Uses:
control of closed environment with UV
(germicidal) lamp.
PHENOL AND PHENOLICS - MOA: disruption of
plasma membrane, denaturation of enzymes.
 Phenol - Seldom used as a disinfectant
or antiseptic because of its irritating
qualities and disagreeable odor.
Uses: rarely used except as a standard
comparison
 Phenolics - Derivatives of phenol that
are reactive even in the presence of
organic material. Example: cresol (O-
phenylphenol). Uses: environmental
surfaces, instruments, skin surfaces, and
mucous membranes
 Biphenols - Broad spectrum, but most
effective against gram-positives.
Example: triclosan; hexachlorophene
Uses: disinfectant hand soaps and skin
lotions
BIGUANIDES - Chlorhexidine is a member of
biguanide group. MOA: disruption of plasma
membrane. Bactericidal to gram-positive and
gram negative. Uses: skin disinfection,
especially for surgical scrubs
HALOGENS - May act alone or as components ALDEHYDES - MOA: protein denaturation. Very
of inorganic and organic compounds. effective antimicrobials
Uses:
 Iodine - MOA: inhibits protein function
and oxidizing agent. Uses: effective  Glutaraldehyde – disinfection of
antiseptic available as tincture and an medical equipment
iodophor  Formaldehyde – excellent disinfectant
 Chlorine - MOA: forms strong oxidizing
GASEOUS STERILANTS - MOA: protein
agent hypochlorous acid that alters
denaturation (Excellent sterilizing agent for
cellular components. Uses: disinfect
agents that can be damaged to heat)
water (gas); disinfect dairy equipment,
Example: Ethylene oxide.
eating utensils, household items and
glassware (chlorine compounds) PEROXYGENS - “Oxidizing agent”.
MOA: oxidation (very effective against oxygen
ALCOHOLS - MOA: protein denaturation and
sensitive anaerobes, contaminated surfaces
lipid dissolution (Bactericidal, Fungicidal, Not
and some deep wounds)
effective against endospores or non-
Uses:
developed viruses). Examples: ethanol and
isopropanol Uses: swabbing the skin with  Ozone – supplement for chlorination
alcohol before injection; thermometer and  Hydrogen peroxide – poor antiseptic
other instruments. but good disinfectant
 Peracetic acid – food processing and
HEAVY METALS AND THEIR COMPOUNDS - MOA:
medical equipment
denaturation of enzymes and other essential
proteins. Examples: silver, mercury, copper
Uses:

 Silver Nitrate – prevent gonorrheal

ophthalmia neonatorum
 Merbromine – disinfects skin and
mucous membranes
 Copper sulfate – algicide

SURFACE-ACTIVE AGENTS

 Soaps - MOA: mechanical removal of

microbes trough rubbing. Uses: skin
degermining and debris removal
 Acid-Anionic Detergents - MOA:
enzyme inactivation or disruption. Non-
toxic, noncorrosive, fast-acting.
Uses: sanitizers in dairy and food
processing industries
 Cationic detergents (Quaternary
ammonium compounds) - MOA:
enzyme inhibition; protein denaturation;
disruption of plasma membrane.
Bactericidal, bacteriostatic, fungicidal
and virucidal against enveloped viruses.
Uses: antiseptic for skin, instruments,
utensils, rubber goods. Ex: benzalkonium
chloride

ORGANIC ACIDS - MOA: metabolic inhibition,

mostly affecting molds (action is not related to
their acidity). Widely used to control molds and
some bacteria in foods and cosmetics
Uses:

 Sorbic acid and benzoic acid –

effective at low pH
 Parabens – cosmetics and shampoos
 Calcium propionate – bread