Topic 10: Control of microorganisms
Aims
• Describe the different physical, chemical, mechanical and biological methods used to
control microorganisms.
Outcomes
• List (Knowledge): Physical, mechanical and chemical methods used for controlling
microorganisms.
• Demonstrate (Application): the mechanisms of microbial control by the different
methods.
• Compare (and contrast) (Analysis): actions of disinfection, antisepsis and sterilisation.
• Distinguish between (Analysis): cidal and static agents.
• Identify the structures of viruses, fungi and bacteria, and their functions,
replication processes
Control of microbial
growth
➢ It is necessary in many practical situations & significant advances in agriculture,
medicine, food science
➢ Control of microbial growth means to inhibit or prevent growth of microorganisms
➢ Physical control: incineration, moist heat like boiling, steam under pressure,
pasteurization, sterilization using the autoclave;
➢ Chemical agents: factors affecting the effectiveness of chemical agents;
➢ canning and commercial sterilization; factors affecting the effectiveness of physical
agents;
➢ Disinfection, antisepsis, sanitization;
➢ Mechanical methods: Filtration
Control of Microbial Growth:
Definitions
• Sterilization: Killing or removing all forms of microbial life
(including endospores) in a material or an object.
• Heating is the most used method of sterilization
• Commercial Sterilization: Heat treatment that kills endospores
of Clostridium botulinum the causative agent of botulism, in
canned food.
• Does not kill endospores of thermophiles, which are not pathogens
and may grow at temperatures above 45oC.
Definitions
➢ Disinfection: Reducing the number of pathogens to a level at which they do not
present a risk to patients or clients
➢ Microorganisms to the point where they no longer cause diseases. Usually
involves the removal of vegetative or non-endospore forming pathogens.
➢ May use physical or chemical methods.
➢ Disinfectant: chemical substance or compound used to inactivate or destroy
microorganisms on inanimate objects.
➢ Antiseptic: antimicrobial substance or compound that is applied to living
tissue/skin to reduce the possibility of infection, sepsis, or putrefaction.
➢ Degerming: Mechanical removal of most microbes in a limited area. Example:
Alcohol swab on skin.
➢ Sanitization: Use of chemical agent on food-handling equipment to meet public
health standards and minimize chances of disease transmission. E.g.: Hot soap
& water.
Terms used for Microbial control
Microbicidal agents : Causes microbial death
❖ Bactericide/bacteriocide
❖ Sporocide
❖ Fungicide
❖ Virucide
Microbistasis: Prevents microbial growth
❖ Bacteriostatic
❖ Fungistatic
Microbial control methods/Controlling microorganisms
Reduce or destroy undesirable microbes in a given area
1. Physical
2. Chemical
3. Mechanical
Mode of action of antimicrobials:
Cellular ➢ Cell wall
Targets of ➢ Cell membrane
➢ Cellular synthetic processes (DNA, RNA)
Control ➢ Proteins
1. Heat – Moist vs Dry
Methods of 2. Cold temperatures
Physical
Control 3. Desiccation
4. Radiation
Physical Agents
1. Heating
Means using of temperature to ensure destruction of all microorganisms
Used for sterilizing Used for
biohazardous waste, glassware,
Used for needles, surgical dressings, metal, and
inoculating glassware, and many types objects that
wires, glassware, of microbiological media, won't melt,
etc liquids, etc etc.
1. Heat (Moist) Moist heat – use of hot water or steam mode
of action
➢ denaturation of proteins
➢ destruction of membranes
➢ destruction of DNA
Sterilization
➢ Steam under pressure
➢ Autoclave
➢ 15 psi/121oC/10-40 min
➢ Steam must reach surface of item being sterilized!
1. Heat (Moist) Pasteurization
➢ heat applied to kill potential agents of infection and spoilage without
destroying the food flavor or value
➢ 63°C - 66°C for 30 minutes
Batch method
➢ 71.6°C for 15 seconds
Flash method
Not sterilization
➢ kills non-spore-forming pathogens and lowers overall microbe count
➢ does not kill endospores or many nonpathogenic microbes
2. Heat (Dry) Dry heat - higher temperatures than moist heat
Incineration
• 600 - 1200oC
• combusts & dehydrates cells
dry ovens
• 150-180oC
• coagulate proteins
Bunsen burner
• 1560oC
• Dehydrates cells and removes water
• Can also sterilize
Thermal Death Measurements
Thermal death time (TDT)
➢ shortest length of time required to kill all microbes at a specified temperature
Thermal death point (TDP)
➢ lowest temperature required to kill all microbes in a sample in 10 minutes
Dry heat (160-180oC, 2-4h); Canning (110-121oC, 25-100 min)
2. Cold Temperatures
➢ Microbistatic refrigeration 0-15oC
➢ slows the growth of microbes
➢ refrigeration 0-15oC
➢ freezing <0oC
➢ used to preserve food, media and cultures
Freezing
Liquid nitrogen (-196oC)
3. Desiccation gradual removal of water from cells
➢ reduced water activity
➢ leads to metabolic inhibition
➢ not effective microbial control
➢ many cells retain ability to grow when water is reintroduced
4. Radiation Ionizing radiation & Nonionizing radiation
➢ deep penetrating power breaks DNA
➢ gamma rays, X-rays, cathode rays
➢ used to sterilize medical supplies & food products
Nonionizing radiation
• little penetrating power to sterilize air, water & solid surfaces
• UV light creates thymine pyrmidines
• interfere with DNA replication
➢ At 250-260nm
➢ Creates AT dimerization resulting in lethal mutations.
Ionizing
Gamma rays and x-rays
Creates free radicals which are toxic
5. Filtration
➢ Gases and liquids can be filter sterilized.
Oxygen tension
➢ Creates anaerobic conditions; thus preventing aerobic growth.
Gases
➢ Fumigation of premises, clothes e.g. Propylene oxide
Sound waves
➢ Sound waves of frequency > 20 000 cycle/second kills bacteria and some viruses
➢ Used to disinfect equipment
Factors which affect the effectiveness of a physical agents
➢ Time or duration;
➢ Temperature;
➢ Microbial load;
➢ Nature of microorganisms;
➢ Previous history of the microorganism;
➢ Age of growth;
➢ pH;
➢ Presence/absence of humidity;
➢ Presence of protective compounds organic material, some ions.
Chemical control
• Disinfectants eg Savlon, ethanol, dettol, hydrogen peroxide etc
• Effectiveness of chemicals
-Time or duration;
-Concentration of disinfectant;
-microbial load; Desirable qualities of chemicals:
➢ rapid action in low concentration
-nature of microorganisms;
➢ solubility in water or alcohol;
-Previous history of microorganisms; ➢ Stable;
➢ broad spectrum, low toxicity;
-Age of growth;
➢ penetrating
-pH; ➢ noncorrosive and non-staining
-Protective compounds e.g organic material, some ions. ➢ affordable and readily available
Chemical Control Of Microbial Agents
➢ Halogens Chemical Control
➢ Phenolics
➢ Chlorhexidine
➢ Alcohols
➢ Hydrogen peroxide
➢ Detergents & soaps
➢ Heavy metals
➢ Aldehydes
➢ Gases and aerosols
Chlorination of water
➢ Hypochlorite
➢ Ozone
➢ Iodine
Preservation of foods
➢ Salt
➢ Sugar
➢ Organic acids
➢ Sulfur dioxide, ethylene oxide
Levels of Chemical Decontamination
High-level germicides
• kill endospores
• devices that are not heat sterilizable and intended to be used in sterile
environments (body tissue)
Intermediate-level
• kill fungal spores (not endospores), tubercle bacillus, and viruses
• used to disinfect devices that will come in contact with mucous membranes but
are not invasive
Low-level
• eliminate only vegetative bacteria, vegetative fungal cells, and some viruses
• clean surfaces that touch skin but not mucous membranes
Biological agents/Antibiotics
Efficiency of Antibiotics
Antibiotics effective against only a limited
range of organisms; narrow-spectrum
antibiotics effective only against gram-
negative bacteria
Standard procedures for
determining antibiotics
sensitivity on test
microorganisms
Class of antibiotics (according to mode of action)
1. -Cell wall inhibitors
Penicillin
Vancomycin
2. -Inhibitors of protein synthesis
Tetracycline
Chloramphenicol
3. -Inhibitors of membrane function
Polymyxin
4. -Nucleic acid inhibitors
Quinolones
5. -Other inhibitors
sulfa drugs
Sterile:
• Inanimate objects free of all life
Sterilization:
• destroys all viable microbes including viruses & endospores
Disinfection:
• destroy vegetative pathogens
• not endospores
Sanitization:
• cleansing technique that mechanically removes microbes to safe
levels
Degerming
• removing organisms from an object’s surface