U1 LAB: CONTROL OF MICROBIAL GROWTH Flashcards
Factors affecting Microbial Growth
- Type of Organism
- Microbial load
- Concentration of agent
- Presence of organic material
- Nature of surface (to be disinfected)
- Contact time
- Temperature
- pH
- Compatibility of disinfectants
This is the causative agent of Mad Cow Disease/CJD and is made up of proteins.
Prions (most resistant, need to burn for it to be killed)
This refers to the absence of microbes in an area or object.
Asepsis
This refers to the methods employed to prevent entry of organisms to body, or equipment used for microbial cultivation.
Aseptic Technique
This involves chemical disinfection of living tissues, as well as in treatment of wounds.
Antisepsis
Chemical agent used in antisepsis
Antiseptic
This is a condition in which microbial growth and multiplication is halted or inhibited.
Bacteriostasis
Bacteriostatic methods include?
refrigeration, dyes
These are chemical sterilants used to destroy all forms of life.
Biocide
Other term for biocide
Chemical sterilants
These are inadvertently introduced to specimens or culture.
Contaminant
This is the process of removal of pathogenic microorganisms so materials are safe to handle/dispose.
Decontamination
This refers to the removal of transient microorganisms from the skin.
Degerming
This is the process of destruction of pathogenic microorganisms except spores, prions, and others.
Disinfection
Degerming methods include?
mechanical cleansing, use of antiseptics
This refers to the reduction of pathogens on eating utensils to safe public health levels.
Sanitization
Sanitization methods include?
mechanical cleansing, use of chemicals
This is the destruction of all forms of life including bacterial spores.
Sterilization
This is the lowest temperature in which all bacteria in culture will be killed in 10 minutes.
Thermal Death Point
This is the minimal length of time in which all bacteria in liquid suspension will be killed at a given temperature.
Thermal Death Time
Methods of control
Physical Means and Chemical Means
Physical Means require the use of?
heat
Chemical Means require the use of?
alcohol
This is the most commonly used physical method.
Heat
2 types of heat
Moist Heat, Dry Heat
Principle of Dry Heat
Oxidation
Examples of Dry Heat
Oven (for glasswares, 3 hours)
Dry Heat
longer exposure time, ___ temperatures
higher
This is used to sterilize glasswares.
Hot Air Oven
Temp requirements for dry heat
160-180C for 1.5-3 hours
Examples for open flame
Bunsen burner, Alcohol lamp
This is used for the sterilization of inoculating loops, needles, mouth of tubes.
Open Flame/Flame sterilization
This is the only way to destroy prions, and is used for disposal of hospital infectious waste.
Incineration (Incinerator)
Incineration is outlawed by?
RA 8749 (Clean Air Act of 1999)
Other term for Moist Heat
Heat under Steam Pressure
Principle of moist heat
Coagulation of Proteins
T/F: Dry heat is more effective than moist heat
False
Moist Heat
_____ time of exposure
shorter
Moist heat is the sterilization method choice for?
heat stable objects
Temp requirements for Autoclaving
121C, 15psi (1 atm) for 15 minutes
This kills all microorganisms except prions.
Autoclave
T/F: In opening the autoclave, you must let it cool down first to decrease pressure inside.
True
This is a form of disinfection under heat that doesn’t kill spores.
Boiling
Temp requirements for boiling
100C for 10-15 minutes
This eliminates food borne pathogens responsible for food spoilage.
Pasteurization
Methods under Pasteurization
- Batch Method
- Flash Method
- High tempt short time
Batch Method
63C for 30 minutes
Flash Method
72C for 15 seconds
High Tempt Short Time
72C-140C then 72C for <5sec
This refers to a flowing steam, is fractional and intermittent.
Tyndallization
Tempt requirements for Tyndallization
100C for 30 mins for 30 days
1st day of Tyndallization
kills vegetative states of bacteria
2nd day of Tyndallization
endospores enter vegetative state
3rd day of Tyndallization
kills endospores in vegetative state
This is used for sterilization of high protein media.
Inspissation
Tempt requirements for inspissation
75-80C for 2 hours on 3 consecutive days
This physical method is used to seperate bacteria from liquids or air.
Filtration
Filtration of liquids is done through:
- Berkefield
- Morton
- Seitz
- Chamberland-pasteur
- Membrane filters
Berkefield uses?
diatomaceous earth
Morton uses?
sintered glass
Seitz uses?
asbestos pad
Chamberland-Pasteur uses?
unglazed porcelain
Membrane filters use?
Cellulose esters
Critical sterilization uses?
0.22um filter
The use of liquid filtration is?
Sterilization of heat sensitive solutions
Filtration of air is through?
HEPA filters
HEPA stands for?
High Efficiency Particulate Air
HEPA filters are able to remove what size of organisms?
> 0.3um
Use of HEPA filters can be seen in?
Laboratory hoods, rooms of immunodeficient patients
This physical method can either be ionizing or non ionizing.
Radiation
MOA of Radiation
alkylation of nucleic acid
Ionizing uses what rays?
Gamma rays / Electron beams
Ionizing
Short wavelength, ______ energy
high
Use of Ionizing radiation
Sterilization of disposable materials
Non Ionizing uses what rays?
UV rays
T/F: UV rays have poor penetration
True
Use of Non ionizing radiation
Sterilization of surfaces
This is a physical method used to remove water to stop metabolic action of bacteria/
Dessication (dehydration)
This is a physical method that uses high concentrations of sugar/salts.
High osmotic pressure
High osmotic pressure can be done through?
Plasmolysis
This is a bacteriostatic physical method.
Exposure to low temperature
This chemical is used for cold sterilization.
Ethylene Oxide
Ethylene oxide utilizes a?
gas chamber with 450-700 mg/L volume
Minimum time for Ethylene oxide
2 hours
Principle of Ethylene oxide
Alkylation of nucleic acids
These gaseous chemicals have a shorter time of killing.
Hydrogen Peroxide and Periacetic Acid
MOA of Alcohol
Protein Denaturation and Lipid Dissolution
Alcohol can penetrate the?
plasma membrane
Examples of alcohol
70% Ethanol
MOA of Aldehydes
Alkylation of Nucleic Acids, Protein Denaturation
Examples of Aldehyde
- 2% Glutaraldehyde
- 40% Formalin
This is a disinfectant under Aldehydes.
2% Glutaraldehyde
This has a bactericidal property under Aldehydes.
40% Formalin
This can be used as an alternative for those who are allergic to Iodine.
Chlorhexidine
MOA of Chlorhexidine
Cytoplasmic Membrane Destruction
Uses of Chlorhexidine
Commonly used antiseptic in surgery
MOA of Halogens
Oxidation of cellular components, Protein denaturation
T/F: Iodine is under Halogens.
True
Forms of Iodine
Tincture or Iodophores
This is the form of iodine that is used for antiseptics, and is composed of alcohol and iodine solutions.
Tincture
This is a form of iodine with an added neutral polymer carrier.
Iodophors
This form of iodine is preferrable, and must be diluted.
Iodophors
This halogen is one of the oldest and common disnifectants.
Chlorine
Chlorine is in the form of what, and is not used as sterilants?
Hypochlorite
Dilution for Chlorine
1:10 dilution of 5.25%
Water Sterilization is done through?
- Addition of Bleach
- Addition of Chlorine
- Collection for Water Testing
Addition of Chlorine is done with what, and where?
Copper Sulfate Pentahydrate, swimming pools and in drinking water
Addition of Bleach
2-3 drops in 1 liter of water for 30 minutes
What is added in collection for water testing?
Sodium Thiosulfate
This chemical method is rarely used, and has a bacteriostatic property.
Heavy Metals
MOA of Heavy Metals
Protein and enzyme inactivation
Mercury
Merthiolate
Silver
1% Silver Nitrate (Crede’s Prophylaxis)
Copper
Copper Sulfate (anti-algae)
Heavy metals
- Mercury
- Sulfur
- Copper
MOA of Oxidizing agents
Oxidation of cellular components
Example of oxidizing agents
Hydrogen peroxide, Ozone
These refer to chemically substituted phenol compounds, less toxic and more effective.
Phenolic Compounds
MOA of Phenolic Compounds
Protein denaturation, cell membrane and enzyme inactivation
Examples of phenolic compounds
Ortho-phenylphenol and ortho-benzyl-para-chlorophenol
Uses of phenolic compounds
Hospital and institutional disinfection
This chemical method is effective against Gr (+), has a residual effect and is toxic.
Hexachlorophene
MOA of Hexachlorophene
interruption of electron transport, enzyme inhibition, membrane destruction
This is effective against Gr (+), and is not affected by organic compounds.
Chloroxylenol
MOA of Chloroxylenol
Cell wall disruption, enzyme inactivation
QUATS stands for?
Quaternary Ammonium Compounds
These are surface active agents which reduce surface tension, and is inactivated by organic compounds and hard water.
QUATS
MOA of QUATS
Protein denaturation, cell membrane destruction
Examples of QUATS
- Benzalkonium Cl (Zephiran)
- Cetylperidium Chloride
QUATS are resistant to?
Mycobacterium tuberculosis
These are used for mechanical action by rubbing and scrubbing.
Soap and Detergents
Soap and detergents possess antibacterial ingredients such as?
Triclocarbon or Triclosan
What is used to check effectiveness of physical methods?
Spore strips (placed before autoclaving)
Spore strips contain the?
endospore
Complete sterilization of spore strips is characterized by?
blackened spore strip
Effectiveness of Physical Methods
Hot-air oven
Bacillus subtilis var. niger
Effectiveness of Physical Methods
Ethylene gas
Bacillus subtilis var. globigii
Effectiveness of Physical Methods
Ionization Radiation
Bacillus pumilus
Effectiveness of Physical Methods
Autoclave
- Geobacillus/Bacillus stearothermophilus
Effectiveness of Chemical Methods
Determination of Phenol Coefficient