FACTORS THAT AFFECT MICROBIAL GROWTH IN VITRO

AVAILABILITY OF NUTRIENTS
 Nutrients are various chemical compounds that all living organisms require in
order to sustain life.
 Organisms will obtain energy from these chemical sources by breaking chemical
bonds.
 Nutrients also serve as sources of carbon, oxygen, hydrogen, nitrogen,
phosphorus, and sulfur as well as other elements (e.g., sodium, potassium,
chlorine, magnesium, calcium, and trace elements such as iron, iodine, and zinc)
that are usually required in lesser amounts.

*About two dozen of the approximately 92 naturally occurring elements are

essential to life.
MOISTURE
 Water is essential for life. Cells are composed of 70% to 95% water.
 All living organisms require water to carry out their normal metabolic processes,
and most will die in environments containing too little moisture.
 However, there are certain microbial stages like bacterial endospores and
protozoan cysts that can survive the desiccation (complete drying process).
TEMPERATURE
 Optimum growth temperature is the temperature at which the organism grows
best.
 When the temperature is below the minimum growth temperature, the
microorganism ceases to grow. When the temperature is above the maximum
growth temperature, the microorganism will die.
 The temperature range at which an organism grows can differ greatly from one
microbe to another and is determined by the enzymes present within the
organism.
 Thermophiles are microorganisms that grow best in high temperature.
 Hyperthermophiles or extreme thermophiles are organisms that favor
temperatures above 100oC.
 Pyrolobus fumarii is an archaeon that can survive in an environment with a
temperature of 113oC.
Temperatures at which various organisms can be found in bodies of water
at Yellowstone National Park
Temperature Organisms
163 F (73 C) or lower
o o Cyanobacteria
144oF (62oC) or lower Fungi
 140oF (60oF) or lower Algae
133oF (56oC) or lower Protozoa
122oF (50oC) or lower Mosses, crustaceans, insects
80oF (27oC) or lower Fish

 Mesophiles are microbes that grow best at moderate temperature. This includes
most of the species that grow on plants and animals and in warm soil and water.
Most pathogens and members of the indigenous microbiota are mesophiles
because they grow best at normal body temperature.
 Psychrophiles prefer cold temperature. These microbes thrive in an
environment that is 130C, has 20% salinity, and contains high concentrations of
ammonia and sulfur.
 Psychotrophs is one group of psychrophiles that has an optimum growth
temperature of 4oC (refrigerator temperature).
 Psychroduric organisms are microorganisms that prefer warm temperatures but
can tolerate or endure very cold temperatures and can be preserved in the
frozen state.
Categories of Bacteria Based on Growth Temperature
Category Minimum Optimum Growth Maximum
Growth Temperature Growth
Temperature ( C)
0 Temperature
(0C) (0C)
Thermophiles 25 50-60 113
Mesophiles 10 20-40 45
Psychrophiles -5 10-20 30

pH
 pH refers to the hydrogen ion concentration of a solution (acidity or alkalinity of
the solution).
 Acidophilic microbes (acidophiles) prefer a pH of 2 to 5. This includes those that
can live in the human stomach and in pickled foods. Fungi also prefer acidic
environment.
 Alkaliphiles prefer an alkaline environment (pH > 8.5). Example of which are
those found inside the intestine (pH approx. 9), in soils laden with carbonate,
and in the so-called soda lakes.
 Vibrio cholerae is a bacterium that causes cholera. It is the only human pathogen
that grows well above pH 8.
Osmotic Pressure and Salinity
 Osmotic pressure is the pressure exerted on a cell membrane by solutions both
inside and outside the cell.
 The ideal situation for a cell to survive is that the pressure inside the cell is equal
to the pressure of the solution inside the cell.
 Osmosis is defined as the movement of a solvent (e.g., water), through a
permeable membrane, from a solution having a higher concentration of solute to
a solution having a lower concentration of solute.
 Hypertonic solutions refer to the greater concentration of solutes outside a cell
than the concentration of solutes inside the cell.
 If the cell is a human cell such as an erythrocyte, the loss of water causes the cell
to shrink; this shrinkage is called crenation, and the cell is said to be crenated.
 If the cell is a bacterial cell, having a rigid cell wall, the cell does not shrink. Instead,
the cell membrane and cytoplasm shrink away from the cell wall (plasmolysis)
and inhibits bacterial cell growth and multiplication.
 Hypotonic solutions refer to the greater concentration of solutes inside the cell
than the concentration of solutes outside the cell.
 If the cell is a human cell such as an erythrocyte, the increased water within the
cells causes the cell to swell (cellular edema). If enough water will enter, the cell
will burst (lyse). Hemolysis is the bursting of erythrocytes.
 If a bacterial cell is placed in a hypotonic solution such as distilled water, the cell
may not burst because of the rigid cell wall, but the fluid pressure within the cell
increases greatly.
 If the pressure becomes so great that the cell ruptures, the escape of cytoplasm
from the cell is referred to as plasmoptysis.
 Isotonic solutions refer to equal concentrations of solutes both inside and outside
the cell. In an isotonic environment, no plasmolysis or plasmoptysis will occur and
the cell has normal turgor (fullness).
 Halophilic organisms are microbes that prefer salty environments such as the
concentrated saltwater found in the Great Salt Lake and solar salt evaporation
ponds. V. cholerae and other Vibrio species are halophilic.
 Haloduric organisms are those that do not prefer to live in salty environments byt
are capable of surviving there such Staphylococcus aureus.
Barometric Pressure
 Most bacteria are not affected by minor changes in barometric pressure. Some
thrive at normal atmospheric pressure (about 14.7 psi).
 Piezophiles thrive deep in the ocean and in oil wells, where the atmospheric
pressure is very high.
Gaseous Atmosphere
 Obligate aerobes prefer the same atmosphere that humans do (about 20%-21%
oxygen and 78%-79% nitrogen, with all other atmospheric gases combined
representing <1%).
 Microaerophiles also require reduced concentrations of oxygen at around 5%.
 Obligate anaerobes are killed by the presence of oxygen.
 Thus, the types and concentrations of gases present in a particular environment
determine which species of microbes are able to live there.
 Capnophiles require increased concentrations of carbon dioxide (usually from 5%
to 10%) to obtain maximum growth in the laboratory.