Module 08: Controlling Microbial Growth in Vitro Flashcards
What are the factors that affect microbial growth?
(1) Availability of Nutrients
(2) Moisture
(3) Temperature
(4) pH (acidity and alkalinity)
(5) Osmotic pressure and salinity
(6) Barometric pressure
(7) Gaseous atmosphere
All living organisms require this to sustain life. They are known as energy sources.
Nutrients
How do organisms obtain energy?
Organisms obtain energy by breaking chemical bonds.
Under moisture, this is known as an essential for life. It is needed to carry out normal metabolic processes.
Water
What are certain microbial stages that can survive the drying process (desiccation)?
(1) Bacterial Endospores
(2) Protozoal Cysts
How does temperature affect microbial growth?
Every organism has an optimum growth temperature. The temperature (and pH) ranges over which an organism grows best are largely determined by the optimum temperature and pH ranges of its enzymes.
These are organisms that grow best at high temperatures.
Thermophiles
These are microbes that grow best at moderate temperatures (eg. 37 degrees Celsius, human body temperature).
Mesophiles (found in warm soil and water like plants and animals)
These are microbes that prefer cold temperatures (like deep ocean water).
Psychrophiles
This is a particular group of psychrophiles, prefer refrigerator temperature (4 degrees Celsius) like bread molds.
Psychrotrophs
These prefer warm temperatures but can endure very cold or even freezing temperatures. They can also be preserved in freezing temperatures.
Psychroduric organisms
This refers to the acidity or alkalinity of a solution.
pH
What pH do most microorganisms prefer?
A neutral or slightly alkaline growth medium (pH 7.0 to 7,4)
These type of microorganisms prefer a pH of 2 to 5. They thrive in highly acidic environments, such created by sulfurous gases in hydrothermal vents and hot springs.
Acidophiles (eg fungi)
These type of microorganisms prefer a pH of >8.5.
Alkaliphiles (such as those in the intestine ph 9 and soda lakes along with soils laden with carbonate)
This is the pressure that is exerted on a cell membrane by solutions both inside and outside the cell.
Osmotic pressure
This is the movement of a solvent, through a permeable membrane, from lower concentration of solutes (dissolved substances) to a higher concentration of solutes.
Osmosis
When the concentration of solutions in the external environment is greater than that of the solutes inside the cell, the solution in which the cell is suspended is said to be what?
Hypertonic
This is a condition in which the cell membrane and the cytoplasm of a cell shrink away from the cell wall.
Plasmolysis (this occurs when a bacteria with a rigid cell walls are placed in a hypertonic solution - so this can inhibit microbial growth and multiplication)
When the concentration of solutes outside a cell is less than that of solutes inside a cell, the solution in which the cell is suspended is said to be what?
Hypotonic
If a bacterial cell is placed into a hypotonic solution, it may not burst. If it does burst, the cytoplasm escapes, this process is known as what?
Plasmoptysis
Why doesn’t the bacterial cell burst when placed in a hypotonic solution?
Because of the rigid cell wall (plants and bacteria)
A solution is said to be this when the concentration of the solutes outside a cell is equal to the concentration of solutes inside the cell.
Isotonic (normal turgor)
Organisms that prefer to live in salty environments are called what?
Halophilic organisms (vibrio cholerae)
Organisms that do not prefer to live in salty environments but are capable of surviving there called what?
Haloduric organisms (Staphylococcus aureus)
Microbes that can survive in high atmospheric pressure (<14.7 psi) are known as what?
Piezophiles
These type of microorganisms prefer the same atmosphere that humans do (~20%–21% O2 and 78%–79% N2, and <1% other gases).
Obligate aerobes
These type of microorganisms require reduced concentrations of oxygen (~5% O2).
Microaerophiles
These type of microorganisms are killed by the presence of oxygen.
Obligate anaerobes
These types of microorganisms require increased concentrations of CO2 (5%–10% CO2).
Capnophiles
This is known as the increase of number of organisms rather than the increase in their size.
Bacterial Growth (growth refers to multiplication and proliferation)
Bacteria divide by ____________ (one cell divides to become two cells) when they reach their optimum size.
Binary fission (to produce two daughter cells after DNA replication)
Binary fission will continue in what conditions?
(1) Binary fission continues through many generations until a colony is produced on solid culture medium.
(2) Binary fission continues for as long as there is a sufficient supply of nutrients, water, and space.
The time it take for one cell to become two cells is called what?
Generation time (eg. E coli, Staphylococcus spp., and Streptococcus spp.. = 20 minutes)
These are used in microbiology laboratories to culture or grow bacteria.
Media or medium
Media prepared in laboratories to culture bacteria are referred as what? This is because they do not occur naturally.
Artificial media or synthetic media
This type of medium is one in which all ingredients are known; this is because the medium was prepared in the laboratory by adding a certain number of grams.
Chemically defined medium (culture media can be liquid or solid).
This is a broth or solid containing a rich supply of special nutrients that promote the growth of fastidious organisms, for example, chocolate agar.
Enriched Medium
This has added inhibitors that discourage growth of certain organisms while allowing the growth of a desired organism, for example, PEA agar
Selective Medium
This medium permits the differentiation of organisms that grow on the medium, for example, MacConkey agar.
Differential Medium
What are some various categories of media that are not mutually exclusive?
(1) Blood Agar is enriched
(2) Differential Medium
This medium is a popular liquid medium in bacteriology laboratories; it supports the growth of all categories of bacteria from obligate aerobes to obligate anaerobes.
Thioglycollate (THIO) broth
Explain the nature of Thioglycollate (THIO) broth.
There is a concentration gradient of dissolved oxygen in the tube; organisms grow only in that part of the broth where the oxygen concentration meets their needs.
This is a selective and differential medium. Lactose fermenters are pink; nonlactose fermenters are white.
MacConkey agar
This is a selective and differential medium. S. aureus is able to ferment mannitol changing the color of the medium from red to yellow.
Mannitol Salt Agar
Obligate anaerobes will grow where there is?
0% oxygen
Microaerophiles will grow where there is?
5% oxygen
Obligate aerobes will grow where there is?
20% to 21% oxygen
These are inoculated with clinical specimens (i.e., specimens collected from patients with a suspected infectious disease).
Culture Media
This involves adding a portion of a specimen to the medium. This is accomplished using a sterile inoculating loop.
Inoculation
This is an approached practiced when it is necessary to exclude microbes from a particular area (e.g., when inoculating culture media).
Aseptic Techniques
Unwanted organisms are referred to as ___________; the growth medium or plate is said to be contaminated.
Contaminants
What should be done before inoculation?
(1) The sterility of the media must be maintained before inoculation.
(2) Avoid touching the surface of the agar.
This minimizes contamination and protects the laboratorian.
Inoculating media within a biologic safety cabinet
What happens when the media is inoculated?
After media are inoculated, they must be placed into an incubator which will maintain the appropriate atmosphere, temperature, and moisture level; the process is known as incubation.
This is the process by which the media will be placed into a incubator which will maintain the appropriate atmosphere, temperature, and moisture level.
Incubation
What are the three (3) types of incubators?
(1)A CO2 incubator (contains 5%–10% CO2)
(2) A non-CO2 incubator (contains room air)
(3) An anaerobic incubator (the atmosphere is devoid of oxygen)
This incubator contains 5%–10% CO2.
CO2 incubator (It is important to keep in mind that a CO2 incubator contains oxygen (about 15%–20%) in addition to CO2. Thus, a CO2 incubator is not an anaerobic incubator.)
This incubator contains room air; thus it contains about 20% to 21% of oxygen.
A non-CO2 incubator
This incubator contains an atmosphere that is devoid of oxygen.
An anaerobic incubator
How do you determine the bacterial population counts?
Microbiologists sometimes need to know how many bacteria are present in a particular liquid at a given time (e.g., to determine bacterial contamination of drinking water).
One can determine either the total number of bacterial cells or the number of viable (living) cells.
This can be used to determine growth by measuring the turbidity of the medium.
Spectrophotometer (As bacteria increase in number, the liquid
becomes turbid (cloudy) and less light passes through.)
This is used to determine the number of viable bacteria in a liquid sample by making serial dilutions of the liquid and inoculating onto nutrient agar; after overnight incubation, the number of colonies is counted.
Viable plate count
This for any particular species of bacterium may be determined by growing a pure culture of the organism in a liquid medium at a constant temperature.
Population growth curve