MICROBIAL GROWTH AND NUTRITION

Prof. Cheryl Lyn C. Ayuste LPT., M.A.

MICROBIAL GROWTH
 Growth takes place on two conditions
 cell synthesizes new cell components and increases in size
 number of cells in the population increases
 Growth of most microorganisms occurs by binary fission.
 Cell division and chromosome replication are coordinately regulated.
MICROBIAL GROWTH
Binary Fission - asexual form of reproduction
- growth of an individual cell continues until cell divides into two new
cells; a cell divides into two, two to four, four to eight, etc.
MICROBIAL GROWTH
Microbial growth is affected by
two major factors:

 Environmental: temperature, pH,

Osmotic conditions

 Chemical: Proper concentrations of

C, H, O, N, P, S, some trace elements,
and some organic cofactors
 MICROBIAL GROWTH
Population Growth - can be measured through the increase of the number
of microbial cells or increase in the number of microbial mass.
- is studied by analyzing the growth curve of a
microbial culture

Growth Rate – change in cell number or cell mass per unit time

Generation - interval for the formation of two cells from one

Generation time – time required for the formation of two cells from one
 MICROBIAL GROWTH

Generation or doubling time

 The time required for a complete fission cycle
 Each new fission cycle or generation increases the population by a factor of 2

 As long as the environment is favorable, the doubling effect continues at a constant rate
 The length of the generation time- a measure of the growth rate of an organism
 Average generation time- 30 to 60 minutes under optimum conditions
 Can be as short as 10 to 12 minutes
This growth pattern is termed exponential.
MICROBIAL GROWTH CURVE

The four phases of the growth curve are identified on the curve.
MICROBIAL GROWTH CURVE: LAG PHASE
 no immediate increase in cell number occurs however cells
in the culture are synthesizing new components.

Variety of reasons for Lag Phase

 cells may be old and depleted of ATP,
 cofactors and ribosomes must be synthesized before
growth can begin.
medium may be different from the one the microorganism
was growing in previously
new enzymes would be needed to use different nutrients
possibly the microorganisms have been injured and
require time to recover.
Whatever the causes, eventually the cells begin to
replicate their DNA, increase in mass, and finally divide.
MICROBIAL GROWTH CURVE: EXPONENTIAL OR
LOG PHASE  microorganisms are growing and dividing at the
maximal rate
 genetic potential,
 nature of the medium
 environmental conditions.

growth rate is constant

completing the cell cycle and doubling in number at
regular intervals
population is uniform in terms of chemical and
physiological properties
exponential phase cultures are usually used in
biochemical and physiological studies
MICROBIAL GROWTH CURVE: EXPONENTIAL OR
LOG PHASE  balanced growth
 all cellular constituents are manufactured at constant rates
relative to each other.

 Unbalanced Growth - change in nutrient level or

environmental condition
 rates of synthesis of cell components vary relative to one
another until a new balanced state is reached

 Shift-up - culture is transferred from a

nutritionally poor medium to a richer one
 there is a lag while the cells first construct new ribosomes to
enhance their capacity for protein synthesis

 Shift-down - culture is transferred from a rich

medium to a poor one
 there is a lag in growth because cells need time to make the
enzymes required for the biosynthesis of unavailable
nutrients
MICROBIAL GROWTH CURVE: EXPONENTIAL OR
LOG PHASE
 Once the cells are able to grow again,
balanced growth is resumed and the culture
enters the exponential phase.

These shift-up and shift-down experiments

demonstrate that microbial growth is under
precise, coordinated control and responds
quickly to changes in environmental
conditions.
MICROBIAL GROWTH CURVE: EXPONENTIAL OR
LOG PHASE
 MICROBIAL GROWTH CURVE: STATIONARY PHASE
 population growth eventually ceases and
the growth curve becomes horizontal
 total number of viable microorganisms
remains constant
 result from a balance between cell division
and cell death or
population may simply cease to divide but
remain metabolically active
 MICROBIAL GROWTH CURVE: STATIONARY PHASE
Reasons for Stationary Phase
 nutrient limitation
 accumulation of toxic waste products
 response to starvation for survival
 morphological changes such as endospore
formation
 decrease somewhat in overall size
 protoplast shrinkage
 nucleoid condensation
 MICROBIAL GROWTH CURVE: STATIONARY PHASE
The more important changes during starvation
are in gene expression and physiology.

Starving bacteria frequently produce a variety

of starvation proteins which make the cell much
more resistant to damage
 increase peptidoglycan crosslinking
 cell wall strength
 Dps (DNA-binding protein from starved cells) protein
protects DNA
 MICROBIAL GROWTH CURVE: STATIONARY PHASE
starved cells become harder to kill
More resistant to
 Starvation
 damaging temperature changes
 oxidative and osmotic damage
 toxic chemicals such as chlorine.

 These changes are so effective that some

bacteria can survive starvation for years.
 MICROBIAL GROWTH CURVE: STATIONARY PHASE
 There is even evidence that Salmonella enterica
serovar typhi and some other bacterial
pathogens become more virulent when starved.

 These considerations are of great practical

importance in medical and industrial
microbiology.
MICROBIAL GROWTH CURVE: STATIONARY PHASE
 MICROBIAL GROWTH CURVE: DEATH PHASE
 decline in viable cells following the stationary
phase
 It was assumed that detrimental environmental
changes such as
 nutrient deprivation
 buildup of toxic wastes caused irreparable harm and loss
of viability

That is, even when bacterial cells were

transferred to fresh medium, no cellular growth
was observed.
Because loss of viability was often not
accompanied by a loss in total cell number, it
was assumed that cells died but did not lyse.
MICROBIAL GROWTH CURVE: DEATH PHASE
MICROBIAL NUTRITION
 Uptake of nutrients that are acquired from the environment and are used for growth
and metabolism.
 Microorganisms (or microbes) vary significantly in terms of the source, chemical form,
and amount of essential elements they need.
MICROBIAL NUTRITION: BASIC NUTRIENTS FOR
GROWTH
ATP for cellular processes
Carbon is necessary for the production of
many macromolecules (proteins, lipids, and
carbohydrates)
Oxygen for metabolism
Nitrogen for amino acid synthesis
Sulfur for vitamins, amino acids, structural
stability of proteins
MICROBIAL NUTRITION: BASIC NUTRIENTS FOR
GROWTH
 Phosphorous makes ATP and membranes
Trace elements are used for metabolic
reaction in the cell and cell component
stabilization.
 Ex.
cobalt Co
potassium K
molybdenum Mo
magnesium Mg
manganese Mn
calcium Ca
iron Fe
zinc Zn
MICROBIAL NUTRITION: BASIC NUTRIENTS FOR
GROWTH
Organic growth factors such as vitamins, amino acids, and nucleic acids some growth
factors cannot be synthesized by own cellular processes.
Water - water activity
 HOW MICROORGANISMS OBTAIN NUTRIENTS?
Heterotroph: uses organic
carbon source
Autotroph: uses inorganic
carbon dioxide
Phototroph: uses light as
energy source
Chemotroph: uses
chemical compounds (ie.
glucose)
 Saprobe
 Parasite
MICROBIAL NUTRITION GROWTH NUTRITION
CULTURING MICROORGANISMS
Inoculum introduced into medium (broth or
solid)
 Environmental specimens
 Clinical specimens
 Stored specimens

Culture – refers to act of cultivating

microorganisms or the microorganisms that
are cultivated
CULTURE MEDIA
Growth medium is used to grow
microorganisms

It contains everything bacteria need to

grow outside the body and under
laboratory conditions.

Provides nutrients for the microorganisms

CULTURE MEDIA

Chemically defined - medium is a growth medium suitable for the microorganisms in

which all of the chemical components are known.
Natural- the exact chemical composition is not known is called natural or empirical
culture media.
 Examples- Milk, urine, diluted blood, vegetable juices, meat extracts, beef and tomato juice, blood
etc.

Living – consists of living cells or tissue; strictly for parasitic bacteria and viruses
which cannot be cultures in non living culture
CULTURING MICROORGANISMS
Special Culture Techniques
 Techniques developed for
culturing microorganisms
 Animal and cell culture
 Low-oxygen culture
 Enrichment culture
CULTURING MICROORGANISMS
CULTURE MEDIA
Enriched Media - added nutrient encourages
the growth of microorganisms.
CULTURE MEDIA
Selective Media - Selects form a
microorganism while inhibiting
most others
Phenol Ethanol Agar - is a selective
medium used to cultivate Gram positive
organisms
Deoxycholate Agar - solid bacteriological
growth medium used for isolation of
enteric pathogens.
 CULTURE MEDIA
Differential Media – Allows for the
differentiation of microorganisms based on
action that occurs on the media or a color
change within the media that is based on a pH
change.
Mannitol Salt Agar - The MSA will select for organisms
such as Staphylococcus species which can live in areas of
high salt concentration (plate on the left in the picture
below).
MacConkey Agar - It is designed to selectively isolate
Gram-negative and enteric bacteria and differentiate
them based on lactose fermentation. Lactose fermenters
turn red or pink on McConkey agar, and non-fermenters
do not change color.
 ASSIGNMENT:
Next Synchronous Meeting : Quiz No. 1
Recitation on the Uptake of Nutrients by the Cell

Worksheet 1: Techniques in Measuring Microbial Growth

Worksheet 2: Major Nutritional Types of Microorganisms
Worksheet 3: Manipulating Microbial Growth (Culture Media)

Reading Assignment 1: Latest researches on Microbial Growth

Reading Assignment 2: Latest researches on Bacterial Cultivation