Bacteria nutrition, metabolism

and growth.
Dr. Edet E. Udo PhD
Department of Microbiology
Faculty of Medicine, Kuwait University.
• 1. Gram- positive bacteria contain a thick layer of peptidoglycan (T/F)
• 2. The periplasmic space is present in both Gram-positive and Gram-
negative bacteria ( T/F)
• 3. Bacterial LPS has both antigenic and virulent properties (T/F)
• 4. Staphylococcus is Gram-positive cocci in clusters (T/F)
• 5. A bacterial spore germinates into two daughter cells ( T/ F).

• Match the following

• 1. Gram- positive cell wall contains____________
• 2. Gram positive cocci in chains __________________
• 3. Has no cell wall ________________
• 4. A primary pathogen __________________________
• 5. An opportunistic pathogen _________________________

• E.coli, Klebsiella spp, Streptococcus, Haemophilus influenzae,

Teichoic acid, Vibro cholerae, Mycoplasma,
Bacteria nutrition, metabolism and growth.

• Objectives
• Define nutrition, explain the nutritional
requirements of bacteria.
• Define bacterial growth.
• Discuss the phases of microbial growth and
their relationship to generation time
• Define metabolism and the fundamental
differences between anabolism and catabolism
• Describe cultivation of bacteria in the
laboratory
 Bacterial nutrition

• Definition:
• Nutrition
• Nutrition is a process by which organisms
acquire chemical substances (Nutrients) used
in cellular activities such as metabolism and
growth.
Organisms differ in the use of particular
elements, their source and chemical form.
Microbial growth
Microbial growth refers to both the increase in
cell size and number of cells in a population
Metabolism
includes all the biochemical reactions
that occur in the cell.
It consists of anabolic and catabolic
reactions.
• Categories of essential nutrients
• Macronutrients: required in relatively
large quantities e.g. proteins,
carbohydrates
• Micronutrients or trace elements:
required in smaller amounts.
• E.g. zinc, manganese.
• Are involved in enzyme functions and
maintenance of protein structure.
 Bacterial growth
Cell division:
– binary or transverse
division.
• During binary fission,
– the parent cell enlarges,
duplicates its
chromosomes and forms
a central transverse
septum that divides the
cell into two daughter
cells.
 Cell division
Generation or doubling time:
The average generation time for
bacteria is 30-60 minutes under
optimum conditions.

Most pathogens such as Staphylococcus

aureus and Escherichia coli double in 20
– 30 minutes.

The longest generation time requires

days. E.g. Mycobacterium leprae that
causes leprosy doubles in 20 to 30
days.
Planes of division
The growth curve
 The growth curve
• The growth curve is a graphic representation
of closed population of bacteria overtime
• This occurs in four phases, lag, log, stationary
and decline phases.
• 1. The lag phase,
• Cells adjust to new environment.
• There is no change in the number of cells but
metabolic activity is high leading to increase in
cellular components.
• 2. The log or exponential phase,
• Bacteria multiply at the fastest rate possible
under the conditions provided.
• Are susceptible to cell wall active antibiotics
• Form metabolic end products
• 3. The stationary phase
• there is an equilibrium between cell division
and cell death caused by :
• decrease in nutrient,
• increase in cell population and accumulation of
metabolic waste / end products e.g
antibiotics.
• Sporulating cells initiate spore formation
• 4. Death or Decline phase
• The number of death cells exceeds the
number of new cells formed due to lack of
nutrients and accumulation of toxic waste
 Factors affecting bacterial
• growth
Nutritional requirements
• Chemical: water, carbon source, nitrogen, minerals,
oxygen, growth factors
• Carbon Source:
• All bacteria require carbon for growth.
• Bacteria can been classified on the basis of their
carbon source.
– Autotrophs: use carbon dioxide as the sole
source of carbon
– Heterotrophs: use more complex organic
compounds such as carbohydrates and amino
acids as source of carbon
– Photoautotroph- energy need is supplied by
light
– Chemoautotroph - energy is extracted from
inorganic substances
• Inorganic nutrients (ions):
– Contain no carbon and hydrogen atoms :
phosphates, potassium, magnesium,
nitrogen, sulfur, iron and numerous trace
metals.
• Organic nutrients:
• contain carbon and hydrogen atoms. Include
carbohydrates, lipids amino acids, Nucleic
acids etc.
• Carbohydrates:
• are used as the initial carbon source for many
biosynthetic pathways and as electron donors
(energy source) by many bacteria.
• Amino acids
• are important source of carbon and nitrogen.
 Nutritional factors
• Phosphorus
• is present as phosphates salts.
• They function in energy metabolism and as
constituents of nucleic acids, phospholipids,
teichoic acids, ATP, etc
• Minerals:
• K, Mg, Ca, Fe are required in relatively high
levels.
• Function as cofactors in enzyme reactions and
as cations they act as buffers within the cells
• Vitamins,
• purines and pyrimidines ( accessory
growth factors) function as coenzymes
 Physical factors
• Oxygen requirement:
• Bacteria can be divided into five groups on the
basis of their oxygen requirements
• 1. Obligate or strict aerobes:
• The growth of bacteria is inhibited by absence
of oxygen An example of a strict or obligate
aerobe is Pseudomonas aeruginosa
• 2. Obligate anaerobes:
• Growth is inhibited by the presence of
oxygen.
– Examples of obligate anaerobe are Clostridium spp
and Bacteriodes spp.
• 3. Facultative anaerobes
• are able to grow in the presence or absence
of molecular oxygen. E.g. include
staphylococci, streptococci, Enteococci, etc.
• 4. Microaerophilic bacteria:
• grow best under increased carbon dioxide
tension. Examples include Neisseria
gonorrhoeae, Haemophilus influenzae
• 5. Aerotolerant bacteria:
• can survive (but not grow) for a short period
of time in the presence of atmospheric
oxygen
• Tolerance to oxygen is related to the ability
of the bacterium to detoxify superoxide and
hydrogen peroxide produced as bye products
of aerobic respiration.
• 1. Superoxide dismutase,
• which converts superoxide ( a toxic
metabolite) into hydrogen peroxide is present
in aerobic and aerotolerant bacteria but not in
obligate anaerobes.

• 2. Catalase,
• which converts hydrogen peroxide into water
and oxygen is also present in all aerobic
bacteria but is lacking in aerotolerant
organisms. Strict anaerobes lack both
enzymes.

• O2- + O2- -superoxide dismutase H2O2

(Hydrogen peroxide) + O2-
• H2O2 + H2O2 Catalase-------2H2O +O2
• Temperature:
• There are three critical temperature ranges for
growth:
• (a) Minimum temperature-
• (b). Maximum temperature:
• ©. Optimum temperature:
• Psychrophiles:
• Has optimum temperature below 15 C but
capable of growth at 0 C
• Mesophiles:
– grow at a range of 20 –40 C. Include most
bacterial pathogens with optimum temp. at
37 C
• Thermophiles:
– microbes that has optimum temperature
above 45 C with a general range of 45-80 C
• Most thermophiles form spores e.g. Bacillus
steareothermophilus
• pH:
• Optimum pH for most bacteria is near pH 7.0
(pH 6.5- pH 7.5)
• Bacteria can be classified as alkalinophiles,
neutrophiles or acidophiles according to their
degree of tolerance to pH changes
 Osmotic pressure:
• When a microbial cell is in a
hypertonic solution cellular
water moves out of the cell
through the cell membrane to
the hypertonic solution.
• This osmotic loss of water
causes shrinkage of the cell
PLASMOLYSIS
• In a hypotonic solution such as
in distilled water, water will
enter the cell and the cell may
be lysed by such treatment
(PLASMOPTYSIS).
• Halophiles
• require high salt concentrations
for growth. Some bacteria can
tolerate 15% salt. E.g. S. aureus
 Bacterial metabolism
• Metabolism consists of catabolic and anabolic
reactions.
– Anabolic reactions are energy-requiring
subset of metabolic reactions, which
synthesize large molecules from smaller
ones.
– Catabolic reactions are the energy –
releasing subset of metabolic reactions,
which degrade or breakdown large
molecules into smaller ones.
• Metabolism is best considered in three stages
– 1. Energy metabolism
– 2. Respiratory metabolism
• 1. Energy metabolism
– Energy used by bacteria is primarily
produced by fermentative and/ or
respiratory metabolic pathways.
• Metabolic reactions are catalyzed by enzymes
– Anaerobic metabolism (fermentative
metabolism
– A. Glycolytic ( Embden-Meyerhof )
pathway
– The major glucose utilization pathway.
– Glycolysis- the metabolism of glucose to
yield pyruvic acid and 2 ATP molecules
– B. Entner-Doudoroff pathway
• Aerobic metabolism ( respiration)
• The Krebs cycle
• Involves the the metabolism of 2-carbon
• Biosynthetic pathways
– Used to build small molecules with nitrogen,
sulphur and other minerals into amino
acids, purines, pyrimidines, polysaccharides
and lipids
 Bacterial metabolism
• Uses of energy
• For biosynthetic activities e.g. cell wall
synthesis, protein synthesis, nucleic acid
synthesis etc
• Membrane transport and movement e.g active
transportation.
• Uses of metabolic products in the
laboratory
• Metabolic end products e.g. pyruvic acid, lactic
acid, mixed gases, alcohols, are used for
bacterial identification. (Biochemical tests) e.g.
oxidase tests, catalase tests.
 Cultivation of bacteria in the
laboratory
• Culture media:
• Any material prepared for the
growth of bacteria in the
laboratory.
• They can be broth (liquid) or
• solid media ( contains a
gelling substance such as agar)
• Microbes that grow on a culture
medium are known as a culture.
• When grown on solid media, the
growth is called colony .
• Growth in a liquid medium is
demonstrated by turbidity.
 Types of Culture media:
• Defined synthetic medium:
– contains known quantities of specific
nutrients
– contain nutrients of reasonably well-
– Complex medium:
– known composition that varies from batch
to batch
• Enrichment medium: (liquid medium)
– is used to encourage the growth of a
particular organism in a mixed culture
• Enriched media: (Solid medium,)
– contain additional nutrients to support the
growth of fastidious organisms. E.g blood
agar and chocolate agar
• Chocolate agar contains lysed blood.
• The lysis releases intracellular nutrients such as haemin (X factor) and
the coenzyme nicotinamide adenine dinucleotide (NAD or V factor)
into the agar for use by fastidious microorganisms such as
Haemophilus species and Neisseria gonorrhoeae.

• Selective media:
• contains salts, dyes or other chemicals that inhibit the growth
unwanted microorganisms.
• Differential media:
• contain chemicals that allow the distinction between different types of
organisms e.g. Lactose in MacConkey agar.

• Preservation of bacterial cultures:

• By refrigeration,, lyophilization, storage in liquid nitrogen
Culture media