CONTROL OF MICROBES IN THE ENVIRONMENT 2.

Hospitals – must be scrupulous in their control of

microorganisms
Terminology of Microbial Growth: 3. Microbiology Laboratories – must use rigorous methods of
controlling microorganism
1. Antiseptic – reduction in the number of microorganisms and
4. Foods and Food Production Facilities – e.g heat treatment,
viruses, particularly potential pathogens, on living tissue
irradiation, chemicals
2. Aseptic – refers to an environment or procedure free of
5. Oher Industries – product quality or safety; cosmetics,
pathogenic contaminations
deodorants, etc.
3. –cide and –cidal – suffixes indicating destruction of a type of
microbe Microbial Death Rates:
4. Degerming – removal of microbes by mechanical means
5. Disinfection – destruction of most microorganisms and Microbial Death – the permanent loss of reproductive ability under
viruses in nonliving tissue ideal environmental conditions
6. Pasteurization – use of heat to destroy pathogens and
Microbial death rate – a technique for evaluating the efficacy of an
reduce the number of spoilage microorganisms in foods and
antimicrobial agent; constant overtime for any particular MQs
beverages
under a particular set of conditions
7. Sanitization – removal of pathogens from objects to meet
public health standards Action of Antimicrobial Agents:
8. –statis and –static – suffixes indicating inhibition, but not
complete destruction, of a type of microbe 1. Alteration of cell walls and membranes
9. Sterilization – destruction of all microorganisms and viruses 2. Damage to proteins and nucleic acids
in or on an object The Selection of Microbial Control Methods:
“Sterility” – is such an absolute term 1. Inexpensive
Decontamination – treatment used to reduce the number of 2. Fast-acting
disease-causing microbe to a level that is considered safe to handle 3. Stable
4. Harmless
Preservation – the process of inhibiting the growth of
microorganisms in products to delay spoilage Factors Affecting the Efficacy of Antimicrobial Methods:

Situational Considerations: 1. Site to be treated

2. Time (length of exposure)
1. Daily life – e.g washing and scrubbing with soaps and 3. Number of MQs
detergents 4. Presence of extraneous matter
 5. Proper exposure Most Susceptible to Most Resistant:
6. Concentration of antimicrobial agent
7. Environmental conditions Enveloped viruses – Gram positive bacteria – Non enveloped viruses
8. Temperature – Fungi – Gram negative bacteria – Active stage protozoa
(trophozoites) – Cysts of protozoa – Mycobacteria – Bacterial
9. pH
endospores
The Selection of Antimicrobial Control Procedure:
Most Resistant to Less Resistant:
1. Type of Microorganism (bacterial endospores of Bacillus
and Clostridium; waxy cell walls of Mycobacterium species, Prions – Endospores of bacteria – Mycobacteria – Cysts of protozoa
Pseudomonas, naked viruses) – Vegetative Protozoa – Gram negative bacteria – Fungi, including
most fungal spore forms – Viruses without envelopes – Gram
2. Numbers of MQs initially present (it takes more time to kill
a large population of bacteria than it does to kill a small positive bacteria – Viruses with lipid envelopes
population) Methods for Evaluating Disinfectant and Antiseptics:
3. Environmental Conditions (e.g pH, temperature, presence
of fats and organic materials, dirt, grease, blood and other a. Phenol Coefficient
fluids) - The ratio that compares a given agent’s ability to
4. Potential Risk of Infection (critical items: needles, scalpels, control microbes to that of phenol under
biopsy forceps; semi-critical items: GIT endoscopes and standardized conditions
endotracheal tubes; non-critical: countertops, stethoscopes, - A phenol coefficient greater than 1.0 indicates that
blood pressure cuffs) an agent is more effective than phenol, and the
larger ratio, the greater the effectiveness
Relative Susceptibility of MQs: b. Use-Dilution Test
- Dips several metal cylinders in broth cultures of
 Classification of Germicides
bacteria and briefly dries at 37°C
a. High-level germicides
- The most effective agent is the one that entirely
- Kill ALL pathogens including bacterial endospores
prevent microbial growth at the highest dilution
b. Intermediate-level germicides
- The current standard test
- Kill fungal spores, protozoan cysts, viruses and
c. Disk-Diffusion Method (Kirby-Bauer Method)
pathogenic bacteria, but not bacterial endospores
- Demonstrates the effectiveness of disinfectants and
c. Low-level germicides
antiseptics through the zone of inhibitions (an area
- Eliminate vegetative bacteria, fungi, protozoa, and
where bacteria do not grow)
some viruses
- Used to disinfect items that only contact the skin of
patients, such as furniture and electrodes
 b. Autoclaving (Pressure Cooking)
- 15mins at 121°C
- Autoclave : Sterilization of medical and laboratory
supplies that can tolerate heat and moisture;
Pressure cooker: sterilization of canned food
c. Pasteurization
- 15sec at 72°C
- Destruction of all pathogens and most spoilage
microbes in dairy products, fruit juices, beer, and
d. In-Use Test wine.
- Swabs are taken from actual objects and are then d. Ultrahigh-temperature sterilization
inoculated into appropriate growth media then are - 1-3sec at 140°C
examined for microbial growth - Sterilization of dairy products
Physical Methods of Microbial Control (1/2) Dry Heat
Health Related Methods a. Hot air
a. Thermal Death Point - 2h at 160°C or 1h at 171°C
- The lowest temperature that kills all cells in a broth - Sterilization of water-sensitive materials such as
in 10 minutes powders, oils, and metals
b. Thermal Death Time b. Incineration
- The time it takes to completely sterilize a particular - 1sec at more than 1000°C
volume of liquid at a set temperature - Sterilization of inoculating loops, flammable
c. Decimal Reduction Time (D) contaminated medical waste, and diseased
- The time required to destroy 90% of the microbes in carcasses
a sample B. stereothermophilus – preferred for monitoring steam sterilization
Moist Heat B. pumilus – most resistant to gamma radiation
B. subtilis – most resistant to ethylene oxide and dry heat processes
a. Boiling
- 10mins at 100°C Physical Methods of Microbial Control (2/2)
- Disinfection of baby bottles and sanitization of a. Refrigeration
restaurant cockware and tableware - 0-7°C
- Preservation of food
b. Freezing
- <0°C
- Long-term preservation of foods, drugs, and
cultures
c. Desiccation (drying)
- Varies with amount of water to be removed
- Preservation of food
d. Lyophilization (freeze drying)
- -196°C for a few minutes while drying
- Long-term storage of bacterial cultures
e. Filtration
- Filter retains microbes
- Sterilization of air and heat-sensitive ophthalmic
and enzymic solutions, vaccines, and antibiotics Chemical Methods of Microbial Growth (1/2)
f. Osmotic Pressure
a. Phenol (carbolic acid)
- Exposure to hypertonic solutions
- Original surgical antiseptic, now replaces by less
- Preservation of food
odorous and injurious phenols
g. Ionizing radiation (electron beams, gamma rays, X-rays)
b. Phenolics (chemically altered ohenol; bisphenols are
- Seconds to hours of exposure (depending on
composed of a pair of limited phenolics)
wavelength of radiation)
- Disinfectants and antiseptics
- Sterilization of medical and laboratory equipment
c. Alcohols
and preservation of food
- Disinfectants, antiseptics, and as a solvent in
h. Nonionizing Radiation ( ultraviolet light)
tinctures
- Irradiation with 260 nm wavelength radiation
d. Halogens (iodine, chlorine, bromine, and fluorine)
- Disinfection and sterilization of surfaces and of
- Disinfectants, antiseptics, and water purification
transparent fluids and gases
e. Oxidizing Agents (peroxides, ozone, and peracitic acid)
- Disinfectants, antiseptics for deep wounds, water
purification, and sterilization of food-processing and
medical equipment
f. Surfactants (soaps and detergents)
- Soaps; degerming; detergents; antiseptics
Biguanides (Chlorhexidine)
- Widely used as an antiseptic in soaps and lotions,
and more recently impregnated into catheters and
surgical mesh
Peroxygens (Hydrogen peroxide and Peracetic acid)
- Hydrogen peroxide is used to sterilize containers for
aseptically packed juices and milk
- Peracetic acid is widely used to disinfect and
sterilize medical devices
Phenolic Compounds (Triclosan and Hexaclorophene)
- Triclosan is used in variety of personal care
products, including toothpaste, lotion, deodorant
soap
- (Biphenolics) Lysol (Triclosan)
- Hexaclorophene is highly effective against
Staphylcoccus aureus, but its use is limited because
it can cause neurological damage; causes brain
damage in infants; absorbed through the skin; not
advisable for pregnant women
Quats
- Are cationic detergents that are non-toxic enough
to be used to disinfect food preparation surfaces;
attracted to cell surface
Selecting the Appropriate Germicidal Chemical
1. Toxicity – by nature, germicides are at least somewhat toxic
to humans and the environment
2. Activity in the Presence of Organic Matter – hypochlorite is
easily inactivated by organic matter unlike phenolics
3. Compatibility with the Material Being Treated – e.g
electrical equipment, metals and rubbers
4. Residue – can be toxic and corrosive
5. Cost and availability
6. Storage and stability
7. Environmental risk – germicides must be neutralize before
disposal
Halogens – tincture of iodine and iodophores can be used as
disinfectants or antiseptics
Chemical Methods of Microbial Growth (2/2)
a. Heavy Metals (arsenic, zinc, mercury, silver, and copper,
etc.)
- Fungistats in paints; silver nitrate creams; surgical
dressings, burn creams, and catheters; coppers,
algicide in water reservoirs, swimming pools, and
aquariums
b. Aldehydes (glutaraldehyde and formaldehyde)
- Disinfectant and embalming fluid
c. Gaseous agents ( ethylene oxide, pimpylene oxide, and
beta-propiolactone)
- Sterilization of heat and water-sensitive objects
d. Antimicrobics
- Disinfectants and treatment of infectious diseases
Chemical Preservatives:
1. Formaldehyde in shampoos
2. Quaternary ammonium compounds in toothpaste
3. Thimerosal in contact lens solution
4. Phenol derivatives in leather belts
5. Benzoic, sorbic, propionic acids in foods duch as bread, CONTROL OF MICROBES IN THE BODY
cheese and juice
6. Nitrate and nitrite in processed meats Clinical Considerations in Prescribing Antimicrobial Drugs
- Is converted to nitrosamines during frying in hot oil
1. Readily available
or by metabolic activities of intestinal bacteria;
2. Inexpensive
shown to be potent CARCINOGENS
3. Chemically stable
4. Easily administered
5. Nontoxic and non-allergenic
6. Selectively toxic

A bacterial cell wall is composed of a macromolecule of

peptidoglycan composed of NAG-NAM chains that are cross-linked
by peptide bridges between the NAM subunits.

New NAG and NAM subunits are inserted into the wall by enzymes,
allowing the cell to grow. Normally, other enzymes link new NAM
subunits to old NAM subunits with peptide crosslinks.

Spectrum of Action

- Is the number of different kinds of pathogens a drug

acts against
a. Narrow-spectrum drugs
b. Broad-spectrum drugs

Efficacy

a. Diffusion Susceptibility Test (Kirby-Bauer Method)

b. Minimum Inhibitory Concentration (MIC)
- Is the smallest amount of drug that will inhibit
growth and reproductions of the pathogen
- Determine via a broth dilution test (turbidity)
c. Minimum Bacterial Concentration (MBC) Test
- The lowest concentration of drug for which no
growth occurs in the Subcultures
Safety and Side Effects:
a. Toxicity
b. Allergies
c. Disruption of normal microbiota
Characteristics of Chemotherapeutic Agents:
Retarding Resistance
1. Highly toxic to a large number of pathogens.
2. No toxicity to the host.
3. Does not induce the development of antibiotic resistance in
mutant MQs.
4. Will not induce hypersensitivity in the host.
5. Does not interfere with the host’s normal defense
mechanism.
Factors Influencing Therapeutic Value:
a. The susceptibility of the pathogen to the chemotherapeutic
agent.
b. The drugs spectrum of activity
c. Possible adverse reactions to the drug
d. The drugs distribution within the body
e. Metabolism of the drug in the body
f. Duration of treatment
g. Interactions with other drugs the patient maybe taking
The Development of Resistance in Populations
a. New mutations of chromosomal genes
b. By acquiring resistance genes on extra-chromosomal
pieces of DNA called R-plasmids or R-factors via
transformation, transduction, or conjugation.
Multiple Resistance and Cross Resistance
- Multiple resistance is resistance to more than one
drug at a time
- Cross resistance is resistance to one drug, and
confers resistance to similar drugs
1. First, sufficiently high concentrations of the
drug can be maintained in a patient’s body for
long, enough time to kill all susceptible cells and
inhibits others long enough for the body’s
defenses to defeat them.
2. Then, use antimicrobial agent in combination so
that pathogens resistant to one drug will be
killed by second, and vice versa.
Synergism – one drug enhances the effect of the
second drug; e.g penicillin and streptomycin
Antagonistic – interfering with each other
3. Lastly, is to limit the use of antimicrobial agent
to necessary cases.