Bacterial Classification, Anatomy, Nutrition, Growth, Metabolism and
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Transcript Bacterial Classification, Anatomy, Nutrition, Growth, Metabolism and
Bacterial Classification,
Anatomy, Nutrition,
Growth, Metabolism and
Genetics
Classification Systems in the Prokaryotes
1.
2.
3.
4.
5.
6.
Macroscopic morphology
•
Colony appearance & color
•
Texture & size
Microscopic morphology
•
Cell shape, size
•
Staining
Physiological / biochemical characteristics
•
Enzymes
Chemical analysis
•
Chemical compound of cell wall
Serological analysis
1.
Ag/ Ab binding
Genetic and molecular analysis
•
G + C base composition
•
Nucleic acid sequencing and rRNA analysis
G + C base composition
Low G+C Gram-Positive Bacteria
Clostridia
Mycoplasmas
High G+C Gram-Positive Bacteria
Corynebacterium
Mycobacterium
Bacterial Taxonomy Based on
Bergey’s Manual
Bergey’s Manual of Determinative
Bacteriology – five volume resource
covering all known procaryotes
based on genetic information –
phylogenetic
two domains: Archaea and Bacteria
five major subgroups with 25 different phyla
classification
Major Taxonomic Groups of Bacteria
Vol 1A: Domain Archaea
primitive, adapted to extreme habitats and modes of
nutrition
Vol 1B: Domain Bacteria
Vol 2-5:
2 - Phylum Proteobacteria – Gram-negative cell
walls
3 - Phylum Firmicutes – mainly Gram-positive with
low G + C content
4 - Phylum Actinobacteria – Gram-positive with high
G + C content
5 – Loose assemblage of phyla – All gram negative
Species and Subspecies
Species
bacterial
cells which share overall similar pattern of
traits
Subspecies
Strain
or variety
culture derived from a single parent that differs in
structure or metabolism from other cultures of that
species
E. coli O157:H7
Type
subspecies
that can show differences
Bacterial Shapes, Arrangements, and Sizes
Typically described by one of
three basic shapes:
coccus
Spherical
bacillus
Rod
coccobacillus
vibrio
spirillum
Helical, twisted rod,
Spirochete
Bacterial Shapes, Arrangements, and Sizes
Arrangement of cells dependent on pattern of
division and how cells remain attached after
division:
cocci:
singles
diplococci
tetrads
chains
irregular clusters
cubical packets
bacilli:
chains
palisades
Cocci
Bacilli
Bacterial anatomy
Generalized structure of a prokaryotic cell
Appendages: Cell Extensions Flagella
3 parts
filament
long, thin, helical structure
composed of proteins
Hook
curved sheath
basal body
stack of rings firmly anchored
in cell wall
rotates 360o
1-2 or many distributed
over entire cell
Flagellar Arrangements
monotrichous
single flagellum at one
end
lophotrichous
small bunches arising
from one end of cell
amphitrichous
flagella at both ends
of cell
peritrichous
flagella dispersed over
surface of cell, slowest
Fig. 4.4
Movement by flagella
Polar
Rotates counterclockwise
Cell swims forward in
runs
Reverse will stop it
Peritrichous
All flagella sweep
towards one end
Chemotaxis
Internal Flagella Axial Filaments
aka Periplasmic
Endoflagella
Spirochetes
enclosed between cell
wall and cell membrane
of spirochetes
Appendages for Attachment Fimbrae
fine hairlike bristles
from the cell surface
function in adhesion
to other cells and
surfaces
Appendages for Mating Pili
rigid tubular structure
made of pilin protein
found only in Gram
negative cells
Functions
joins bacterial cells for DNA
transfer (conjugation)
Adhesion
to form biofilms and
microcolonies
The Cell Envelope
External covering outside the cytoplasm
Composed of few basic layers:
glycocalyx
cell wall
cell membrane
Maintains cell integrity
The Cell Membrane
fluid layer of phospholipid and protein
phospholipid molecules are arranged in a bilayer
Hydrophobic fatty acid chains in the phospholipids form a
permeability barrier
The Bacterial Surface Coating
Glycocalyx
Coating of molecules
external to the cell wall
Made of sugars and/or
proteins
functions
attachment
inhibits killing by white
blood cells
receptor
The Bacterial Surface Coating
Glycocalyx
2 types:
1. slime layer loosely organized
and attached
2. capsule - highly
organized, tightly
attached
Cell Wall
Four Groups Based on Cell Wall
Composition:
1. Gram
positive cells
2. Gram negative cells
3. Bacteria without cell walls
4. Bacteria with chemically unique cell walls
Structure of the Cell Wall Peptidoglycan
macromolecule
composed of a
repeating framework
of long glycan chains
cross-linked
by short
peptide fragments
provides strong,
flexible support
keep
bacteria from
bursting or collapsing
because of changes
in osmotic pressure
Gram Positive Cell Wall (1)
Consists of
a thick, homogenous
sheath of peptidoglycan
tightly bound acidic
polysaccharides
teichoic acid and
lipoteichoic acid
Periplasmic space
cell membrane
Gram Negative Cell Wall (2)
Consists of
an outer membrane
containing
lipopolysaccharide (LPS)
periplasmic space
thin shell of peptidoglycan
periplasmic space
cell membrane
Protective structure while
providing some flexibility
and sensitivity to lysis
Gram Negative Cell Wall
LPS
endotoxin that may
become toxic when
released during infections
may function as receptors
and blocking immune
response
contains porin proteins in
upper layer
Regulates molecules
entering and leaving cell
The Gram Stain
Important basis of bacterial
classification and identification
Practical aid in diagnosing infection
and guiding drug treatment
Differential stain
Gram-negative
lose crystal violet and stain red from
safranin counterstain
Gram-positive
retain crystal violet and stain purple
Atypical Cell Walls
Some bacterial groups lack typical cell wall
structure
Mycobacterium and Nocardia
Gram-positive cell wall structure with lipid mycolic
pathogenicity
high degree of resistance to certain chemicals and dyes
basis for acid-fast stain
Some have no cell wall
Mycoplasma
cell wall is stabilized
pleomorphic
by sterols
acid
Chromosome
single, circular, doublestranded DNA molecule
contains all the genetic
information required by a
cell
DNA is tightly coiled
around a protein
dense area called the
nucleoid
central subcompartment in
the cytoplasm where DNA
aggregates
Plasmids
small circular, doublestranded DNA
stable extrachromosomal
DNA elements that carry
nonessential genetic
information
duplicated and passed on to
offspring
replicate independently from the
chromosome
Plasmids
may encode antibiotic
resistance, tolerance to toxic
metals, enzymes & toxins
used in genetic engineering
readily manipulated &
transferred from cell to cell
F plasmids allow genetic
material to be transferred from
a donor cell to a recipient
R plasmids carry genes for
resistance to antibiotics
Storage Bodies Inclusions & Granules
intracellular storage
bodies
vary in size, number &
content
Examples:
Glycogen
poly-b-hydroxybutyrate
gas vesicles for floating
sulfur
Endospores
resting, dormant cells
produced by some G+ genera
Clostridium, Bacillus & Sporosarcina
resistance linked to high levels of
calcium & certain acids
longevity verges on immortality
25 to 250 million years
pressurized steam at 120oC for
20-30 minutes will destroy
Endospores
have a 2-phase life cycle
sporulation
formation of endospores
Germination
vegetative cell
endospore
return to vegetative growth
withstand extremes in heat,
drying, freezing, radiation &
chemicals
Endospores
• stressed cell
• undergoes asymmetrical cell division
• creating small prespore and larger
mother cell
• prespore contains:
•
mother cell matures the prespore
into an endospore
•
•
Cytoplasm
DNA
dipicolinic acid
then disintegrates
environmental conditions are again
favorable
protective layers break down
• spore germinates into a vegetative
cell
•
Microbial nutrition,
growth, and
metabolism
Obtaining Carbon
Heterotroph
organism
that obtains carbon in an organic
form made by other living organisms
proteins, carbohydrates, lipids and nucleic
acids
Autotroph
an organism that uses CO2 (an inorganic
gas) as its carbon source
not dependent on other living things
Growth Factors
organic compounds
that cannot be
synthesized by an
organism & must be
provided as a nutrient
essential amino acids,
vitamins
Carbon Energy
source source
photoautotrophs
CO2
sunlight
chemoautotrophs
CO2
Simple
inorganic
chemicals
photoheterotrophs
organic
sunlight
Nutritional types
Chemo
Chemical compounds
Photo
light
chemoheterotrophs organic
Metabolizing
organic cmpds
Types of Heterotrophs
Saprobes
Parasites / pathogens
Obligate
Nutritional Movement
Osmosis
Facilitated diffusion
Active transport
Endocytosis
Phagocytosis
Pinocytosis
Extracellular
Digestion
digestion of complex
nutrient material into
simple, absorbable
nutrients
accomplished through
the secretion of
enzymes (exoenzymes)
into the extracellular
environment
Environmental Influences on
Microbial Growth
1.
2.
3.
4.
5.
6.
temperature
oxygen requirements
pH
Osmotic pressure
UV light
Barophiles
1. Temperatures
Minimum temperature
lowest
temperature that
permits a microbe’s growth
and metabolism
Maximum temperature
highest
temperature that
permits a microbe’s growth
and metabolism
Optimum temperature
promotes
the fastest rate of
growth and metabolism
Temperature Adaptation Groups
Psychrophiles
optimum temperature 15oC
capable of growth at 0 - 20oC
•
•
Mesophiles
optimum temperature 40oC
Range 10o - 40oC (45)
most human pathogens
•
•
•
Thermophiles
optimum temperature 60oC
capable of growth at 40 - 70oC
•
•
Hyperthermophiles
Archaea that grow optimally
above 80°C
found in seafloor hot-water
vents
2. Oxygen Requirements
Aerobe
requires oxygen
Obligate aerobe
cannot
grow without
oxygen
Anaerobe
does
not require oxygen
Obligate anaerobe
Facultative anaerobe and aerobe
capable
of growth in the absence OR
presence of oxygen
Fluid thioglycollate media can be used to
test an organism’s oxygen sensitivity
Gas chamber
3. pH
The pH Scale
Ranges
from 0 - 14
pH below 7 is acidic
pH
pH
[H+] > [OH-]
above 7 is alkaline
[OH-] > [H+]
of 7 is neutral
[H+] = [OH-]
3. pH
Acidophiles
Neutrophiles
optimum pH is relatively to
highly acidic
optimum pH ranges about
pH 7 (plus or minus)
Alkaphiles
optimum pH is relatively to
highly basic
4. Osmotic Pressure
Bacteria 80% water
Require
water to grow
Sufficiently hypertonic media at concentrations
greater than those inside the cell cause water
loss from the cell
Osmosis
Fluid leaves the bacteria causing the
Causes the cell membrane to separate
Plasmolysis
Cell
cell to contract
shrinkage
extreme or obligate halophiles
Adapted
to and require high salt concentrations
5. UV Light
Great for killing bacteria
Damages the DNA
(making little breaks)
in sufficient quantity can kill
the organisms
in a lower range causes
mutagenisis
Endospores tend to be
resistant
can survive much longer
exposures
6. Barophiles
Bacteria that grow at
moderately high hydrostatic
pressures
Barotolerants
Grows at pressures from 100500 Atm
Barophilic
Oceans
membranes and enzymes
depend on pressure to
maintain their threedimensional, functional shape
400-500
Extreme barophilic
Higher than 500
Microbial Associations
Symbiotic
organisms
live in close nutritional relationships;
Mutualism
Obligatory
Dependent
Both members benefit
Commensalism
One member benefits
Other member not harmed
Parasitism
Parasite is dependent and benefits
Host is harmed
Microbial Associations
Non-symbiotic
organisms
are free-living
relationships not required for survival
Synergism
members cooperate and share nutrients
Antagonism
some member are inhibited or destroyed by others
Microbial Associations
Biofilms
Complex relationships
among numerous
microorganisms
Develop an extracellular
matrix
Adheres cells to one
another
Allows attachment to a
substrate
Sequesters nutrients
May protect individuals in
the biofilm
Microbial Growth in Bacteria
Binary fission:
Prokaryotes
reproduce
asexually
one cell becomes two
basis for population growth
Process:
parent cell enlarges
duplicates its chromosome
forms a central septum
divides the cell into two
daughter cells
Population Growth
Generation / doubling time
time required for a complete
fission cycle
Length of the generation time
is a measure of the growth rate
of an organism
Some populations can grow from
a small number of cells to several
million in only a few hours!!
Prokaryotic Growth
Bacterial Growth Curve
•
lag phase
•
•
logarithmic (log) phase
•
•
•
Exponential growth of the population occurs
Human disease symptoms usually develop
stationary phase
•
•
no cell division occurs while bacteria adapt to their new
environment
When reproductive and death rates equalize
decline (exponential death) phase
•
accumulation of waste products and scarcity of resources
Other Methods of Analyzing
Population Growth
Turbidity
Direct microscopic count
Coulter counting
Turbidity
Direct Microscopic Count
Electronic Counting
Microbial genetics
Genomes
Prokaryotic Genomes
Prokaryotic
chromosomes
Main portion of DNA, along
with associated proteins
and RNA
Prokaryotic cells are
haploid (single
chromosome copy)
Typical chromosome is
circular molecule of DNA in
nucleoid
DNA Replication in Prokaryotes
Genetic Recombination in Prokaryotes
Genetic recombination
occurs when an organism acquires
and expresses genes that
originated in another organism
Genetic information in prokaryotes
can be transferred vertically and
horizontally
Vertical gene transfer (VGT)
transfer of genetic material from
parent cell to daughter cell
Horizontal gene transfer (HGT)
transfer of DNA from a donor cell to
a recipient cell
Three types
Bacterial conjugation
Transformation
Transduction
DNA Recombination Events
3 means for exogenous genetic
recombination in bacteria:
Conjugation
2. Transformation
3. Transduction
1.
Transmission of Exogenous
Genetic Material in Bacteria
conjugation
requires the attachment of two
related species & formation of a
bridge that can transport DNA
transformation
transfer of naked DNA
transduction
DNA transfer mediated by
bacterial virus
1. Conjugation
transfer of a plasmid or chromosomal fragment from
a donor cell to a recipient cell via direct connection
Gram-negative
cell donor has a fertility plasmid
(F plasmid, F′ factor)
allows the synthesis of a conjugation (sex) pilus
recipient
cell is a related species or genus
without a fertility plasmid
donor transfers fertility plasmid to recipient
through pilus
F+ and F-
Physical Conjugation
2. Transformation
chromosome fragments from a
lysed cell are accepted by a
recipient cell
genetic
code of DNA fragment is
acquired by recipient
Donor and recipient cells can be
unrelated
Useful tool in recombinant DNA
technology
Transformation of Insulin Gene
human insulin gene isolated and cut from its
location on the human chromosome
using a restriction enzyme
plasmid is cut using the same restriction
enzyme
desired DNA (insulin gene) and plasmid DNA
can be joined using DNA ligase
plasmid now contains the genetic instructions
on how to produce the protein insulin
Bacteria can be artificially induced to take up
the recombinant DNA plasmids and be
transformed
successfully transformed bacteria will
contain the desired insulin gene
transformed bacteria containing the insulin
gene can be isolated and grown
As transformed bacteria grow they will produce
the insulin proteins coded for the recombinant
DNA
Insulin harvested and used to treat
diabetes
3. Transduction
DNA is transferred from one
bacterium to another by a
virus
Bacteriophages
Virus
that infects bacteria
consist of an outer protein
capsid enclosing genetic
material
serves as a carrier of DNA
from a donor cell to a recipient
cell
Other ways genetics can change:
Transposons
Mutations
Transposons
Special DNA segments that have the
capability of moving from one location
in the genome to another
“jumping genes”
Can move from
one chromosome site to anotherr
chromosome to a plasmid
plasmid to a chromosome
May be beneficial or harmful
Changes in traits
Replacement of damaged DNA
Transfer of drug resistance
Mutations
Result of natural
processes or induced
Spontaneous
mutations
heritable changes to the
base sequence in DNA
result from natural
phenomena such as
radiation or uncorrected
errors in replication
UV
light is a physical
mutagen that creates a
dimer that cannot be
transcribed properly
Nitrous acid is a chemical mutagen that converts
adenine bases to hypoxanthine
Hypoxanthine
base pairs with cytosine instead of thymine
Base analogs bear a close resemblance to
nitrogenous bases and can cause replication errors
Point Mutation
Result of spontaneous or
induced mutations
affects just one base pair
in a gene
Base-pair substitutions
result in an incorrect base in
transcribed mRNA codons
Base-pair deletion or
insertion
results in an incorrect
number of bases
Repair Mechanisms
Attempt to correct mistakes or damage in the DNA
Mismatch repair involves DNA polymerase
“proofreading” the new strand
removing mismatched nucleotides
Excision repair
involves
cutting out
damaged DNA
replacing it with
correct nucleotides