CONTROLLING MICROBIAL GROWTH IN THE ENVIRONMENT

Terminologies for Microbial Control

1. Sterilization - removal or destruction of all microbes including viruses and bacterial

endospores, in or on an object.
2. Aseptic - describes an environment or procedure that is free of contamination by
pathogens.
3. Disinfection - the use of physical or chemical agents known as disinfectants to inhibits
or destroy microorganisms, especially pathogens.
4. Sanitization- the process of disinfecting plates and utensils used by the public to reduce
the number of pathogenic microbes to meet acceptable public health standards.
5. Pasteurization- the use of heat to kill pathogens and reduce the number of spoilage
microorganisms in food and beverages. Examples: milk, fruit juice, wine, and beer are
commonly pasteurized.
6. -statis or -static - suffixes indicating inhibition but not complete destruction of a type of
microbe.
7. -cide or -cidal - refers to agents or methods that destroy or permanently inactive a
particular type of microbe.

Microbial Death Rates

- Microbial death as the permanent loss of reproductive ability under ideal environmental
conditions.
- One technique for evaluating the efficacy of an antimicrobial agent is to calculate the
microbial death rate.
- Usually found to be constant over time for any particular microorganism under a
particular microorganism under a particular set of conditions.

Antimicrobial Action
Two basic categories:
1. Disrupt the integrity of cells by altering their cell walls/ cytoplasmic membranes.
2. Interrupt cellular metabolism and reproduction by interfering with proteins and
nucleic acids structure

Alteration of Cell Walls and Membranes

Cell wall: counteracting effects of osmosis when in hypotonic solution

Effects of physical or Chemical agents:

● inability to protect cell from osmosis and or death.
Viral envelope- attachment of virus to target cell:
● Damage prevents viral replication
● Lack of envelope- greeted tolerance of harsh environment and antimicrobials
Damage to Proteins and Nucleic Acids
- Proteins regulate cellular metabolism
- Like enzymes in most metabolic reactions
- Forms structural components in membranes and cytoplasm
● Broken down by extreme heat (or radiation) and chemicals - denatured proteins
● Denatured proteins - cease to function and bring about cell death

The Selection of Microbial Control Methods

- Ideal agents do not exist.
● Inexpensive, fast acting and stable during storage

Factors Affecting the Efficacy of Antimicrobial Methods

- Site to be treated
● Extreme heat can’t be applied on humans and animals
● In medical procedures
● Instruments should be sterilized
● Disinfection - surface of mucus membranes
- Relative Susceptibility of Microorganism
● Microbial death rate is usually constant for a particular agent against a single microbe

Microorganism Susceptibility
● Bacterial endospores - Bacillus and Clostridium - the most resilient forms of life
● Species of mycobacteria: eg. Mycobacterium tuberculosis - waxy lipid- strong
disinfectants and heat
● Cyst of Protozoa - prevents entry of disinfectant, drying and radiation and heat

Prions- are infectious proteins that cause degenerative diseases of the brain, are more
resistant than any cell or virus.

Prion Diseases
● Several conditions fall under the term prion diseases
● A prion is type of protein that can trigger normal proteins in the brain to fold abnormally
● Affect both humans and animals and are sometimes transmitted to humans by infected
meat products

Germicidal effectiveness
● Prions - heating to 482oC for 4 hours to denature
● High- germicides that kill all pathogens, including bacterial endospores
● Intermediate- fungal spores, protozoan cysts,
○ viruses, and pathogenic bacteria, but not bacterial endospores
● Low- eliminate vegetative bacteria, fungi, protozoa and some viruses
○ Disinfect items that contact only skin of patients, such a furniture and electrodes.
Environmental conditions
● Such as temperature and pH
● Warm disinfectants: chemicals react faster at higher temperatures
● Acidic Conditions - enhance the antimicrobial effect of heat
○ Household chlorine bleach, are more effective at low pH

Organic Materials
● Fat, feces,vomit,blood
● Intercellular matrix of biofilms
● Interfere with the penetration of heat, chemicals and some forms of radiation
● In some cases there materials, inactive chemical disinfectants

Biosafety Levels
● CDC guidelines for four levels of safety in microbiological laboratories dealing with
pathogens:
● BSL 1- minimal precautions: Hand Washing
● BSL 2- mod. Hazardous agents; extreme prec to contaminates obj. Or sharps
● BSL3- hepa filter
● BSL4- most secure; sealed airlocks and multiple showers with uv light room

Methods for Evaluating Disinfectants and Antiseptics

(Scientist have developed several methods to measure the efficacy of antimicrobial growths)

● Phenol- is an antiseptic used during surgery in the late 1880s. Calculating a ratio
that compares the agent’s ability to control microbes
● Phenol coefficient - greater than 1.0 indicates that an agent is more effective
than phenol
● Use-dilution test- the most effective agent is the one that entirely prevents
microbial growth at the highest dilution.
● In-use-tests - provide accurate determination of an agents efficacy under real life
conditions

Physical Method of Microbial Control

- Exposure of the microbes to extremes of heat and cold, desiccation, filtration, osmotic
pressure, and radiation.

Heat Related Methods

- High temperatures denature proteins, interfere with the integrity of cytoplasmic
membranes and cell walls, and disrupt the function and structure of nucleic acids.
 - The thermal death point is the lowest temperature that kills all cells in 10 minutes. While
thermal death time is the time that it takes to complete sterilize a particular volume of
liquid at a set temperature.
- Decimal Reduction time (D) is the time required to destroy 90% of the microbes in a
sample.
Moist heat - kills by denaturing proteins and destroying nucleic acids
➔ Boiling- kills the vegetative cells of bacteria and fungi, the trophozoites of protozoa, and
most viruses within 10 minutes at sea level
➔ Autoclaving- pressure is applied to boiling water to prevent the escape of heat in steam
➔ Pasteurization - heating beer and wine to destroy the microorganism that causes
spoilage (not sterilization)
● Brucella melitensis - undulant fever/brucellosis
● Mycobacterium bovis - bovine tuberculosis
● Escherichia coli - diarrhea
● Ultra high temperature sterilization - flash heating milk or other liquids to rid them of
all living microbes
○ Heating beverages at higher temperature than pasteurization
● Dry Heat- powders and oil, cannot be sterilized by boiling or steam; some metal objects,
can be damaged by repeated exposure to stem
● dry heat requires higher temperature for longer times

- Complete incineration - the ultimate means of sterilization ; contaminated

Refrigeration and Freezing

- Food preparation and storage, the most convenient method of microbial control is either:
- Refrigeration (temperatures between 0 degree celsius and and 7 degree celsius)
- Freezing (temperature below 0 degree celsius)

PROCESS:
Decreases microbial metabolism, growth, and reproduction because chemical reactions occur
more slowly at low temperature and because liquid water is not available at subzero
temperatures ( usually mesophiles; except psychrophilic) .

Desiccation and Lyophilization

- Desiccation or drying preserve such as foods as fruits and vegetables, grains, nuts, and
yeasts
- Inhibits the spread of most pathogens, including the bacteria that cause syphilis,
gonorrhea, and the more common forms of bacterial pneumonia and diarrhea
- Lyophilization - freezing and drying, to preserve microbes and other cells for many
years
Filtration
- Filtration - the passage of liquid through a material that traps and removes microbes.
- Filtration traps microbes larger than the pore size allowing smaller microbes to pass
through
- HEPA filters are mounted in the air ducts of some operating rooms, rooms occupied by
patients with airborne diseases such as tuberculosis, and rooms of immunocompromised
patients , such as burn victims and AIDS patients.

Osmotic Pressure
● Use of high concentration of salt or sugar in foods to inhibit microbial growth
● The removal of water inhibits cellular metabolism because enzymes are fully functional
only in aqueous environments
● Fungi have a greater ability than bacteria to tolerate hypertonic environments with little
moisture, which explains why jelly in your refrigerator may grow a colony of Penicillium
mold but is mold likely to grow the bacteria Salmonella

Radiation
● Physical method of microbial control
● Lonizing Radiation-Electron beams, gamma rays and some X-rays, all of which
● Have wavelengths shorter than 1nm
● Nonionizing Radiation-Ultraviolet (UV) light, visible light infrared radiation, and radio
waves
○ UV light has sufficient energy to be a practical antimicrobial agent

Chemical Methods of Microbial Control

● Phenol and phenolics
● Alcohols- —-- bactericidal, fungicidal, and virucidal agents enveloped viruses; however,
they are not effective against fungal spores or bacterial endospores; intermediate- level
disinfectants
○ Alcohols denature proteins and disrupt cytoplasmic membranes
○ Pure alcohol is not effective antimicrobial- denature of proteins requires water
○ 70% to 90% alcohol; control microbes
○ Evaporate rapidly; leave no residue but disadvantage may not contact microbes
long enough to be effective
○ Alcohol based antiseptics; more effective than soap for bacteria on hands but not
effective on some virus; eg. diarrhea- causing noroviruses
○ Swabbing the skin with alcohol prior to an injection removes more microbes
○ By physical action (degerming) then by chemical actin.
● Halogens- iodine; betadine;chlorine; NaOCI; chlorine dioxide- gas;bromine(pools/hot
tubs); fluorine
● Oxidizing Agents - H202- catalase neutralizes it on open wound; ozone; percetic -
sterile medical;equipment
● Surfactants- soaps and detergents
 ● Heavy Metals- silver-wounds; copper-pools
● Aldehydes-formalin-embalming, hospital
● lGaseous Agents - ethylene oxide- hospitals; explosive and poisonous
● Enzymes - prionzymes

● Antimicrobial Drugs

Development of Resistant Microbes

● There is little evidence that the extensive use of such products adds to human or animal
health, but it does promote the development of strains of microbes resistant to
antimicrobial chemicals
● While susceptible cells die, resistant cells remain to proliferate
● Scientific have already isolated trains of pathogenic bacteria, including M. tuberculosis,
P. aeruginosa, E. coli and S. aureus, that are less susceptible to common disinfectants
and antiseptics