Bacterial Nutrition, Growth

and Genetics

May 15, 2008

MD 3
Microbiology
Elem % of dry
Source Function
ent weight
Carb organic compounds
50 or CO2 Main constituent of cellular material
on
H2O, organic
Oxyg Constituent of cell material and cell water; O2 is electron
20 compounds, CO2, and
en acceptor in aerobic respiration
O2

Nitro NH3, NO3, organic Constituent of amino acids, nucleic acids nucleotides, and
14
gen compounds, N2 coenzymes

Hydr H2O, organic

8 Main constituent of organic compounds and cell water
ogen compounds, H2
Phosp inorganic phosphates Constituent of nucleic acids, nucleotides, phospholipids,
3 (PO4)
horus LPS, teichoic acids
Sulfu SO4, H2S, So, organic Constituent of cysteine, methionine, glutathione, several
1
r sulfur compounds coenzymes
Potas Main cellular inorganic cation and cofactor for certain
1 Potassium salts
sium enzymes
Magn Inorganic cellular cation, cofactor for certain enzymatic
0.5 Magnesium salts
esium reactions
Calci Inorganic cellular cation, cofactor for certain enzymes and
0.5 Calcium salts
um a component of endospores
Trace elements

metal ions required by certain cells in

such small amounts that it is difficult
to detect (measure) them
not necessary to add them to culture
media as nutrients
usually act as cofactors for essential
enzymatic reactions in the cell.
Mn, Co, Zn, Cu, and Mo.
All living organisms require a
source of energy.
Phototrophs: use radiant energy/light
as energy source
Heterotrophs: use organic carbon as
the source
Lithotrophs: use inorganic compounds
as energy source
Autotrophs: use CO2 as energy
source
Growth factors are small amounts of
organic compounds which is not
synthesized by the bacteria, but essential
for its growth
1. purines and pyrimidines: required for
synthesis of nucleic acids (DNA and RNA)
2. amino acids: required for the synthesis
of proteins
3. vitamins: needed as coenzymes and
functional groups of certain enzymes
Satellitism: Haemophilus influenzae
and Staphylococcus aureus
Auxotrophs

Mutant strains of bacteria that require

some growth factor not needed by the
wild type (parent) strain
Example:
E. coli trp-.
Common vitamins required in the
nutrition of certain bacteria.
Nicotinic acid
Pantothenic acid
Riboflavin (B2)
Vitamin K
CULTURE MEDIA

isolation and maintenance of pure

cultures of bacteria
identification of bacteria according to
their biochemical and physiological
properties.
Liquid media are used for growth of
pure batch cultures
solid media are used widely for the
isolation of pure cultures, for
estimating viable bacterial populations
usual gelling agent for solid or
semisolid medium is agar
Types of Culture Media

Chemically-defined (synthetic) medium

is one in which the exact chemical
composition is known.

Complex (undefined) medium is one in

which the exact chemical constitution of the
medium is not known.
complex media usually contain complex
materials of biological origin such as blood
or milk or yeast extract or beef extract
Selective medium is one which has a
component(s) added to it which will inhibit
or prevent the growth of certain types or
species of bacteria and/or promote the
growth of desired species.
One can also adjust the physical conditions
of a culture medium, such as pH and
temperature, to render it selective for
organisms that are able to grow under
these certain conditions
Thayer-martin media
 Differential medium distinguishes
between different types of bacteria
based on some observable trait in
their pattern of growth on the medium
An enrichment medium contains some
component that permits the growth of
specific types or species of bacteria,
usually because they alone can utilize the
component from their environment.
Oxygen requirements

Obligate aerobes
-require O2 for growth
Obligate anaerobes
-O2 is a toxic substance, which either
kills or inhibits their growth.
-fermentation, anaerobic respiration,
bacterial photosynthesis, or
methanogenesis.
Facultative anaerobes (or
facultative aerobes)
-fermentation or anaerobic respiration
-aerobic respiration.
Aerotolerant anaerobes
-exclusively anaerobic (fermentative)
type of metabolism
-insensitive to the presence of O2.
 Superoxide Catala Peroxid
Group
dismutase se ase

Obligate aerobes and most facultative

+ + -
anaerobes (e.g. Enterics) 

Most aerotolerant anaerobes (e.g.

+ - +
Streptococci)

Obligate anaerobes (e.g. Clostridia,

- - -
Methanogens, Bacteroides)
always consider the optimum pH for
growth of a desired organism and
incorporate buffers in order to
maintain the pH of the medium
Most diagnostic media for the growth
and identification of human pathogens
have a pH near 7.
Effect of temperature
Psychrophiles: grow at 0 C
Psychrotrophs: grow at 0 C but its
optimum temperature is 37C
Mesophiles: grow at 37 C (body
temperature)
Thermophiles: grow between 45 to
70C
Hyperthermophiles/extreme
thermophiles; grow at 80 to 115 C
Mild halpohiles: require 1-6% salt
Moderate halophiles: require 7-15% salt
Extreme halophiles: require 16-30% salt
Halotolerant: bacteria which grow at
moderate salt concentration, however they
grow best in the absence of salt
Osmophiles : grow in high sugar
environments
Xerophiles: grow in dry environments( lack
or absence of water)
Bacterial Genetics

The genetic material of bacteria and

plasmids is DNA.
Bacterial viruses (bacteriophages or
phages) have DNA or RNA as genetic
material.
The two essential functions of genetic
material are replication and
expression.
Most bacteria have a haploid
genome:
single chromosome
circular, double stranded DNA
linear chromosomes have been found
in Gram-positive Borrelia and
Streptomyces spp.
PLASMIDS

Plasmids are extrachromosomal

genetic elements in bacteria.
smaller than the bacterial
chromosome.
encode traits that are not essential for
bacterial viability
replicate independently of the
chromosome.
PLASMIDS

supercoiled, circular, double-stranded

DNA molecules, but linear plasmids
have also been demonstrated in
Borrelia and Streptomyces.
Conjugative plasmids

code for functions that promote

transfer of the plasmid from the donor
bacterium to other recipient bacteria
F plasmids
promote transfer of the bacterial
chromosome from the donor
bacterium to other recipient bacteria
Copy number:average number of
molecules of a given plasmid per bacterial
chromosome
control medically important properties of
pathogenic bacteria
resistance to one or several antibiotics
production of toxins
synthesis of cell surface structures required
for adherence or colonization
R plasmids
Plasmids that determine resistance to
antibiotics
Representative toxins encoded by
plasmids:
heat-labile and heat-stable enterotoxins
of E coli
exfoliative toxin of Staphylococcus
aureus
tetanus toxin of Clostridium tetani.
BACTERIOPHAGES
bacterial viruses or phages
infectious agents that replicate as obligate
intracellular parasites in bacteria
extracellular phage particles are
metabolically inert
consist principally of proteins plus nucleic
acid (DNA or RNA).
protective shell (capsid) surrounding the
tightly packaged nucleic acid genome
ECLIPSE PERIOD: immediately after infection
during which intracellular infectious phage cannot
be detected.
LATENT PERIOD: interval from infection until
extracellular progeny start to appear
RISE PERIOD: interval from the end of the latent
period until all phage are extracellular.
BURST SIZE: The average number of phage
particles produced by each infected cell; is
characteristic for each virus and often ranges
between 50 and several hundred.
VIRULENT PHAGES: production of phage
progeny and death of the host bacteria.

Lysogeny is a specific type of latent viral infection

in which the phage genome replicates as a
prophage in the bacterial cell
production of infectious phage does not occur
immune to superinfection by the virus which they
harbor as a prophage.
TEMPERATE PHAGES
Some temperate phages contain
genes for bacterial characteristics that
are unrelated to lytic phage
development or the lysogenic state
expression of such genes: phage
conversion (or lysogenic conversion).
production of diphtheria toxin by
Corynebacterium diphtheriae.
TRANSFORMATION
uptake and integration of naked DNA
from the environment into the host
cell
homologous recombination with the
chromosome of the host.
Need nucleases to recognize and
breakdown these foreign DNA.
Haemophilus and certain Bacillus
LOG PHASE
Transformation can be induced
artificially in the lab.
Incubation of E.coli at 4C, in the
presence of calcium ions, then a short
exposure to 42C
HOMOLOGY: pieces of DNA can only
recombine with the chromosome if
there is a high degree of nucleic acid
similarity
TRANSDUCTION
Generalize: phage will pick up any portion
of the bacterial chromosome at random
( not specific for a particular chromosomal
segment).
penicillinase gene of Staphlococci is
located in a plasmid, and the phages can
pick up that plasmid and through
transduction, pass it on to another cell,
thus transferring the penicillanase gene to
other staphylococci species.
SPECIALIZED OR RESTRICTIVE
TRANSDUCTION
phages have a specific insertion site
on the chromosome and can pick up
only a short length of DNA containing
a few genes on either side of this
site.
CONJUGATION
This is the most important
mechanism for widespread
transfer of genetic information
between bacteria.
direct transfer of bacterial DNA
between organisms
requires CELL-TO-CELL CONTACT.
Most conjugation is PLASMID-
MEDIATED: F plasmid
In a very small proportion of cells, the
F plasmid is incorporated into the
bacterial chromosome.
the entire bacterial chromosome acts
like a F plasmid.
 high frequency recombination strains
(Hfr strains).
This leads to 2 processes:
1. The F plasmid and the entire
bacterial DNA undergoes conjugation
with an F(-) cell.
2. The F plasmid is excised along with a
portion of the bacterial DNA, hence
the F plasmid is termed F prime (F’)
TRANSPOSONS
segments of DNA that can move from one
site in a DNA molecule to other target sites
in the same or a different DNA molecule
independent of generalized recombination.
cause mutations and transfer new genes
interrupts the linear sequence of a gene;
inactivates
cause deletions, duplications, and
inversions of DNA segments
not self-replicating genetic elements