CONTROL OF
MICROBIAL GROWTH
 I. INTRODUCTION
A.    IMPORTANCE
• 1.  prevent and control infectious diseases in
humans, animals and plants.
• 2.  preserve food.
• 3.  prevent contaminating microbes from
interfering with certain industrial processes.
• 4.  prevent contamination of pure culture

B.   MEANS OF CONTROL
• 1.  Destroy or inhibit disease-causing microbes.
• 2.  Block the source, route and vector of transmission of
agents.
• 3. Protect an infected person from the consequences of
disease by building up body’s defenses and administration
of appropriate chemotherapeutic drugs.
DEFINITION OF TERMS

1.       STERILIZATION
         - Complete destruction of all living organisms
         - Include cells, viable spores and viruses
         STERILE---devoid of microbial life
         - Done through heat, autoclave, gas, chemicals and
radiation

2.       DISINFECTION
         - Destruction or removal of pathogens only from
nonliving objects by physical or chemical methods.
3.       – CIDAL and –STATIC
         -cidal------killing agents
         -static-----inhibits growth and reproduction

4.       ASEPSIS - absence of pathogens on living tissues

            SEPSIS - growth of pathogens on living tissues

5.       TECHNIQUES
         Aseptic technique  remove all pathogens
         Sterile technique  remove all microorganisms
     FACTORS AFFECTING EFFECTIVENESS OF
ANTIMICROBIAL PROCEDURES

1.     Length of time
2.     Temperature
3.     Concentration
4.     Nature and number of microbes and spores
present (BIOBURDEN)
5.     Presence of organic matter
Ex. Feces, blood, vomitus and pus
 II. PHYSICAL ANTIMICROBIAL METHODS
A.     HEAT
·  Advantages: Practical, efficient and
inexpensive
·   Factors: Temperature and Time
·
THERMAL DEATH POINT (TDP)
- Lowest temperature that kill all
organisms w/in a specified period of time

THERMAL DEATH TIME (TDT)

- Length of time needed to sterilize at a
specified temperature .
• 1.       MOIST HEAT
-- Kills microorganisms by protein coagulation
-- Method and Effectivity:
a. Boiling—10 mins. at 70 C
b. Steaming—10 to 30 mins. at 90 to 100 C
-- Heat resistant microbes
a. Bacterial endospores
(e.g. Anthrax, Tetanus, Gas Gangrene, Botulism)
b. Mycobacteria
c. Viruses (e.g. Hepatitis)
--Affected by altitude (longer boiling time)

• TYNDALIZATION/FRACTIONAL STERILIZATION
-- Endospores of bacteria are destroyed
by boiling and cooling 3x
2. PRESSURIZED STEAM (AUTOCLAVE)
-- Uses steam under pressure
• -- inc Pressure: dec temperature: dec time
• -- Effectivity: 15 psi at a temperature
of 121.5 C for 20 mins.
• -- Destroys all microbial life

3. DRY HEAT
• -- Kills by oxidation effects
METHODS:
a. Flaming - hold end of the loop in the yellow portion
of a gas flame
b. Incineration (Burning) -- destroy contaminated
disposable materials
c. Hot Air Sterilization —Effectivity: 160 - 165 C for
2 hours OR 170 - 180 C for 1 hour
• 4. PASTEURIZATION
• --Mild heating to kill the organism w/o seriously damaging
the taste of the product
--At least 72 C for only 15 secs. OR 62 C for 30 secs.

B. COLD
        --Cold temperature and freezing
        -- MICROBIOSTATIC METHOD - slow metabolic
activities

C. DRYING (DESSICATION)
       --Water is removed that makes microbes unable
to grow or reproduce but can remain viable

       --LYSOPHILIZATION (FREEZE-DRYING)
--Microbes are frozen at temperature -54 to –72 C
and the water is removed by high vacuum
 D.   RADIATION
1.  IONIZING RADIATION
--Gamma rays, x-rays or high electron beams
--Radiation  ionized water  highly reactive
hydrogen  radicals  destruction of DNA
--Not widespread in routine sterilization
--Used for sterilizing pharmaceutical, medical, dental
supplies and food processing plants
2.  NON-IONIZING
--Ultraviolet (UV) light (240 to 280 nm)
Ex. Sunlight, UV lamp
--Damages DNA (form thymine dimmers)
--Used to control microbes in air
--MAJOR DISADVANTAGES:
a.  Organisms to be killed must be directly exposed
b.  Damage to human eyes
c. Burns and skin cancer (prolonged exposure)
 E. ULTRASONIC WAVES
--Used for delicate equipment
--Sound waves mechanically dislodge organic
debris on instruments and glassware

F. FILTRATION
--Filters, separates cells, larger viruses, bacteria
and certain microbes from liquid or gases
--Cellulose ester membranes
--Pore size: 0.22 micro removes all EXCEPT
Chlamydia, Mycoplasma, Rickettsiae, viruses
--FACTORS OF EFFECTIVENESS:
1. Concentration of chemical
2. Contact time
3. pH of Solution
4. Temperature
 III. CHEMICAL ANTIMICROBIAL METHODS

· Use of chemical agents to inhibit the growth of

microorganisms, either temporarily or permanently

· Characteristics of Good Chemical Antimicrobial Agent

1. Kill pathogens within reasonable period and in
specified concentrations
2. Non-toxic, non-corrosive and non-destructive
3. Water soluble and easy to apply
4.  Inexpensive and easy to prepare
5.  Stable in dissolved or solid form
6. Stable to pH and temperature changes
A. AGENTS THAT DAMAGE THE CELL MEMBRANE
• - Release of small metabolites and interference of
active transport and energy metabolism

• 1. Surface Active Disinfectants (SADS)

A.  Quaternary Ammonium Compounds (quats)
-- Cationic agents
-- Bactericidal to gram (+) species
- Antibacterial activity is reduced in presence of
organic matter
        --E.g. Benzalkonium Chloride (Zephiran)
       Cetyl pyridinium Chloride (Cepacol)
     B. Soap and Detergents
         -- Anionic agents dissociate to yield a (-) ion
-- most effective against gram (+)
2. Phenolic Compounds
--Rapidly bactericidal at low concentration
a. Cresols—Ex. Lysol
b. Diphenyl Compound—bactericidal at high
concentration
-- Ex. Hexachlorophene used in germicidal soaps and
antiperspirants
3. Alcohol
--Skin disinfectant (not for instrument
sterilization)
--Active against gram (+), gram (-) and AFB
--Most effective at 50 to 70% concentration
-- Ex. Aliphatic alcohol (Ethanol), isopropyl alcohol
A. AGENTS THAT DENATURE PROTEINS
--Acids, alkalis, alcohol, acetone
--Causes unfolding of polypeptide chainchains
become
randomly and irregularly coiled
--Used as preservatives (benzoic, lactic, Acetic, Citric)
B. AGENTS THAT MODIFY FUNCTIONAL GROUPS OF
PROTEINS AND NUCLEIC ACIDS
-- Bactericidal activity decreased by organic
substances
1. Heavy Metals - soluble salts of Hg, Ag, As and
other metals
a. Mercurials
- Useful antiseptic agents
- Unreliable skin disinfectant
--Ex. Metaphen, Merthiolate, Mercurochrome
b.  Silver Compounds
- Bactericidal
- Silver Nitrate 1% - prophylaxis for opthalmia
neonatorum (Credes prophylaxis)
--Silver sulfadiazine -- burns
2.  Oxidizing agents
a. Halogens
- Bactericidal and effective against sporulating
organism
- Iodine - skin disinfection (superior to other
agents)
- Chlorine - water disinfectant Ex. Chlorox
b.  H2O2
- Weak Antiseptic
- For cleansing of wounds
- Brief germicidal action
3.  Dyes
·   Stain bacteria and inhibitory at very high
dilutions
·   Basic dyes are the most effective
·   Inactivated by serum and other CHONS
·   Usually used in dermatologic treatment
--Ex. Triphenylmethane Dyes, Acridine dyes
4.  Alkylating Agents
· Bactericidal in all including spore formers
a. Formaldehyde
--Formalin - preserving fresh tissue
--As a gas - decontaminate rooms, buildings, fabrics
and instruments
b. Ethylene Oxide
-- Used in gaseous sterilization
c. Glutaraldehyde
-- Cold sterilant for surgical instruments
-- 10x more bactericidal and sporocidal than
formaldehyde
IV. CHEMOTHERAPEUTIC TREATMENT
Antimicrobial agents
--- chemotherapeutic agents used to treat
infectious diseases

Ideal Antimicrobial agents

A.     Kill or inhibit growth of pathogens
B.     Cause no damage to host
C.     Cause no allergic reaction in host
D.     Be stable when stores in solid or liquid form
E.     Remain in specific tissues
F. Kill pathogens before they mutate or
become resistant
A.  ANTIBACTERIAL AGENTS
· Some are bacteriostatic, some are bacteriocidal

1. Inhibition of Cell Wall Synthesis

 Penicillin, Cephalosphorin, Bacitracin, Vancomycin

2. Inhibition of Protein Synthesis

A. 30s subunit---Aminoglycoside, Tetracycline
B. 50s subunit---Chloramphenicol, Lincomycin,
Erythromycin, Clindamycin

3. Disruption of cell membrane—Polymyxin

4. Inhibition of bacterial enzyme activity

 Sulfonamides, Trimethoprim

5. Inhibition of nucleic acid---Quinolone

B. ANTIFUNGAL AGENTS
1.Bind with cell membrane sterols
(Ex. Nystatin, Amphotericin B)
2. Interfere with sterol synthesis
(Ex. Clotrimazole, miconazole)
3. Block Mitosis or nucleic acid (Ex. Griseofulvin)

C. ANTIPROTOZOAL AGENTS
1. Interfere with DNA and RNA synthesis
Ex. Chloroquine, pentamidine, and quinacrine
2. Interfere with protozoal metabolism
Ex. metronidazole

D. ANTIVIRAL AGENTS
-- Inhibits viral replication within the cell
Ex. Acyclovir, valaciclovir, amantadine,
zidovudine, lamivudine, saquinavir