UNIVERSITY INSTITUTE OF PHARMA

SCIENCES
Pharmaceutical Microbiology
20PST-292

Introduction to Microbiology
Dr. Gaurav Joshi
GRAPHY DISCOVER . LEARN . EMPOWER
Microbiology - The science that studies very small living things
Usually requires a magnification tool – the microscope
Some organisms are large though – Helminths – worms
Sub groups of Microbes we will study
Bacteria
Archaea
Fungi
Protozoans
Algae
Viruses
Multicellular animal parasites – Helminths
Bacteria
Fungi
 Protozoans
Giardia Ameba
 Viruses
Bacteriophage Avian Flu
 Helminth
Tapeworm Ascaris round worm
Various disciplines of study within microbiology:
Bacteriology, Mycology, Parisitology, Immunology,
Epidemiology,
Biotechnology
Virology
Environmental Microbiology
Bioremediation
Historical review of the Science of Microbiology
Robert Hook – 1665 – Englishman, used a primitive compound (two magnifying lenses)
microscope, reported that life’s smallest units were little boxes – Cells, his work
started the process of the development of the Cell theory of life
Hook’s microscope
Antoni Van Leeuwenhoek –1673 - probably the first person to observe living cells with
a simple microscope, amateur scientist, ground his own lenses and described what we
know today as bacteria – rod shaped , spiral shaped , etc. “animalcules”
Antoni Van Leeuwenhoek
Francesco Redi – 1668 – opposed the prevailing theory of
Spontaneous Generation,
maggots in meat , He used covered jars to show that maggots came
from flies –strong evidence against spontaneous generation
Now we teach the theory of Biogenesis – Life comes from Life
But issue of Spontaneous Generation was actively believed for many
more years
Francesco Redi
Edward Jenner is credited with first vaccine – in epidemics of smallpox during the late
1700’s he observed that milk maids didn’t get the disease, cattle had a similar
disease – cowpox, milk maids had cow pox lesions, but not small pox, he purposefully
took scrapings from cowpox blister and scraped a 8 year old volunteer. With the
material – child got mild illness but not small pox,

Vaccination comes from Latin word “vacca” meaning cow. Jenner laid the foundation
for Pasteur’s later work with other vaccinations.
(Vaccinia virus similar to Variola smallpox virus, today a possible bioterror weapon –
genetically engineered variola?)
Jenner and vaccination
Pasteur – French sceintist that dealt the death blow to the spontaneous generation
theory.

He devised the ingenious curved necked flasks that prevented contaminated air from
reaching boiled beef broth – the broth remained uncontaminated even though
exposed to the air

He was very lucky – no endopores present, or it would have failed

(resitant to boiling)
1. He developed process we call Pasteuriztion – he heated wine to kill
contaminating microbes – cured sick wine (today we heat treatment to kill
pathogens in milk also)
2. He proved that fermentation was caused by a microbe – yeast
3. He developed vaccines for rabies and anthrax. Vaccines led to immunity
to diseases that routinely killed many people, used to help people long
before they understood how they even worked (science of Immunology)
4. He began the revolution in science that led to the Golden Age of
Microbiology (from 1857-1914)
Robert Koch - Developed Koch’s postulates – important technique for determining the
actual microbial cause agent of a disease – more later, German, contemporary of Pasteur, several
very important contributions
1. He discovered the tuberculosis bug (tubercle bacillus, Mycobacterium tuberculosis)
2. He discovered the cause of anthrax (Bacillus anthracis) – from blood of dead cattle, cultured
bacteria in pure culture, injected bacteria in live cattle and they died, then again cultured the
bacteria in pure culture. This led to the
establishment of a procedure for determining microbial cause of disease (see p. ____-for
modern application of Koch’s postulates)
Koch’s postulates
Koch’s and Pasteur’s work helped establish the “Germ Theory
of Disease” - that
microorganisms cause disease (in people, animals, and even
plants)
Iwanowski 1892 - Discovered that plant disease can be caused by small
organisms that
were so small they passed through filters , Tobacco mosaic virus (TMV) was
later
identified as the cause - beginning of virology (Today we have discovered
new and
weird things like viroids, prions) See next slide
Microbiology – Chapter 1
Paul Ehrlich, German doctor, wanted to find a “magic bullet” an agent that
would kill
the disease agent without hurting the patient (see the “Coming plague”
video, part 1)
1. Developed Salvarsan, “salvation from syphilis”agent
2. This was an arsenical – arsenic compound, that was effective against
syphilis
Antimicrobial agent, medicine to treat a microbial disease, it was
chemical – chemotherapy
• Syphilis spirochete: T. pallidum Paul Ehrlich
Joseph Lister – 1860’s, English surgeon that applied ideas of the germ
theory to surgery, remembered the work of Semmelweis in Hungary in the
1840’s, if a Dr. would wash their hands childbed fever was prevented. He
knew that phenol would kill bacteria; put 2 and 2 together and treated
wounds with phenol - and no infection (phenol is the basic agent of
today’s – Lysol)
1. First antiseptic use in surgery, chemicals used as agents on tissue
before surgery
(tissue treated with an antimicrobial agent – antiseptic, betadine)
disinfectants are chemicals, used on a surface
2. Also proved that microbes cause surgical infections (today’s scourge
–MRSA)
Semmelweis
Lister
• Antisepsis in surgery
Alexander Fleming - Scottish physician and bacteriologist - 1928
Observed mold growing on a bacteria culture, there was a ring of clearing around the mold where
the bacteria didn’t grow, the mold was later found to be a Penicillium species and the naturally
secreted chemical was called penicillin, an antibiotic
1. Antibiotics are natural agents
2. Synthetic drugs are chemicals produced in labs (sulfas)
3. Problems with them - toxicity, resistance, allergic reactions
4. Fleming’s work - shelved until early WWII, sulfas were failing, needed penicillin to cure battle
field wounds
5. Now have thousands of antibiotics and synthetics (and a significant problem – resistance)
Flemming and Penicillium
 Salk - Polio vaccine, 1950’s polio was a scary epidemic, Salk developed a
vaccine by
treating the virus with formalin (IPV) inactivated polio virus
Sabin 1963 live Polio virus vaccine, attenuated –altered virus, OPV-oral
polio vaccine
The work done on polio revolutionized the science of virology and we are
seeing
the results today in advances with Hepatitis and HIV viral infections - tissue
culture and other techniques
Avery and Macleod, DNA is genetic material
Watson and Crick, DNA, 1953
DNA Double helix

• Double Helix
Salk; IPV
Sabin, OPV
 Microbiology – Chapter 1
Jacob and Monod – 1965 Did research on RNA and protein synthesis in bacteria - last
necessary step in understanding how genetics works on a cellular level (Replication,
Transcription, Translation – protein synthesis – expression of traits)
Modern science thrives today only on the laid foundation of thousands of men and
women who did mundane routine and often boring lab science. Don’t forget that we
are
here today because we stand on the shoulders of Giants – who were people just like
you
and me. (Who are the Giants today? CDC, USAMRID, young Joe or Mary the graduate
student?, maybe you?)
Microbiology – Chapter 1
Microbiology – Chapter 1
• Classification, Taxonomy, Binomial Nomenclature - Yucchy!!!!! No
one likes it. Just do it!!!
The Classification of Living Things

• Living organisms are assigned to groups based upon their similarities.

• Systematics is the discipline of identifying and classifying

organisms.
Domains
• The highest – largest category, recent addition
• 3 domains
• 1. Archaea – ancient “bacteria”, unicellular like bacteria, also simple cell
structure (prokaryote – no nucleus) but have distinct metabolism (chemistry)
allowing them to exist in “extreme” environments
• 2. Bacteria – unicellular, prokaryote, found everywhere (Old kingdom name
– Monera)
• 3. Eukarya – unicellular to multicellular, complex and organized cells with
nuclei and organelles (mitochondria)
Domain Archaea
• Archaea are single- celled
organisms that lack a membrane-
bound nucleus. - Prokaryote

• Archaea can be found in

environments that are too hostile
for other life forms.
Domain Bacteria
• Bacteria are single- celled
organisms that lack a membrane-
bound nucleus. (Prokaryote also)

• Bacteria are found almost

everywhere on the planet Earth.
Domain Eukarya
• The cells of all eukaryotes have a membrane-bound nucleus.
Members of the Domain Eukarya are further categorized into one of
four Kingdoms. (know these kingdoms)
Microbiology
• Check your notes; Older 5 kingdom scheme is still widely used
• Monera – bacteria (Prokaryotic)
• Protista – Protozoans (Eukaryotic)
• Fungi - yeast, molds, etc. (Eukaryotic)
• Plant – photosynthetic producers (Eukaryotic)
• Animals – heterotrophic consumers (Eukaryotic)
Categories of Classification
Domain
Kingdom
Phylum
Class
Order
Family
Genus
Species
Categories of Classification
Domain - Dumb
Kingdom - King
Phylum - Philip
Class – Came
Order - Over
Family - For
Genus - Good
Species - __
Spaghetti?
Categories of Classification

Domain Most inclusive

Kingdom
Phylum
Class
Order
Family
Genus
Species
Least inclusive
Categories of Classification
Scientific Names
• Binomial (two name) Genus first and first letter capitalized, then
species not capitalized. If written or typed – either underline or
italicize
• Genus name, species name
• Homo sapiens - italicized
• Homo troglodytis (Your EX?) – underlined
• Canis familiaris – Your puppy (except poodles)
• Felis domesticus – Your Kitty (some strange people have to have Felis leo or
Felis tigris)
• Canis latrans – Invites your kitty over for lunch
• Canis lupus - ____________? guess
Know for test
• 3 domains Archaea, Bacteria, Eukarya
• 4 Eukarya Kingdoms
• Protista – Protozoans like Ameba and Paramecium (producers and consumers)
• Fungi – yeasts, molds, mushrooms (consumers)
• Plants – complex producers, trees, grasses
• Animals – complex consumers
Classification of Bacteria
• Somewhat different: a clinical rapid ID is often important when trying to
find causative agent of a disease
• Bergey’s manual: Manual is in lab for a reference when doing unknown
• Developed on 1940’s for grouping bacteria according to standard
diagnostic lab techniques available at the time (such as Grams stain)
Classification of Bacteria
• Gram + cocci Gram - bacilli
Classification of Bacteria
• Gram - Spirochete Gram + bacilli
Classification of Bacteria
• The manual divides bacteria into 4 groups or divisions on the basis of
their Cell Wall
• 1. Gram + (stain violet)
• 2. Gram - (destain, and are counterstained pink or reddish color)
• 3. Bacteria that lack a cell wall (mycoplasma)
• 4. organisms that have a cell wall lacking “peptidoglycan” (archaeobacteria –
Now called “Archaea”
Classification of Bacteria
• More modern methods now used:
• 1. DNA studies, genomics, gene probes
• 2. using Bacterial viruses “bacteriophages”
• 3. Serology – antibody – antigen reactions (like blood typing)

• Examples: Salmonella typhi

• Salmonella typhimurium
• Azotobacter vinelandii 12837
• E. coli 0157H7 nasty strain of E. coli
 STRUCTURE OF BACTERIA
• Smaller and simpler in structure than eukaryotic cells, with no
recognizable organelles.
• All of the activities performed by organelles also take place in
bacteria, but they are not carried out by specialized structures.
• The small size, simple design, and broad metabolic capabilities
of bacteria allow them to grow and divide very rapidly and to
inhabit and flourish in almost any environment.
 STRUCTURE OF BACTERIA
• They were first seen under a microscope by Anton van
Leeuwenhoek in 1676.
• As microscopes have improved, scientists have come to
understand bacterial cell structure better.
 Bacterial cell structure
• organized into 3 categories :
• Internal Structures: Cytoplasm, nucleoid, bacterial
chromosome, plasmid, ribosomes, and storage granules
• Cell envelope: cell membrane, peptidoglycan cell wall or an outer
lipid membrane (only found in Gram-negative cells)
• External structures (appendages & coverings): flagella,
fimbriae, sex pilus and glycocalyx
 Intracellular structures
• Cytoplasm
• Chromosome
• Plasmid
• Ribosomes
• Inclusion bodies
 Cytoplasm
• Portion of the cell that lies within the PM
• substances within the plasma membrane, excluding the genetic
material.
• Gel-like matrix composed of mostly water(4/5 th ), enzymes,
nutrients, wastes, and gases
• Contains cell structures - ribosomes, chromosome, and plasmids , as
well as the components necessary for bacterial metabolism.
• It is relatively featureless by electron microscope - although small
granules can be seen.
• carries out very important functions for the cell -
growth, metabolism, and replication .
 Constituents

– Proteins including enzymes

– Vitamins
– Ions
– Nucleic acids and their precursors
– Amino acids and their precursors
– Sugars, carbohydrates and their derivatives
– Fatty acids and their derivatives
 Nucleoid
• Unlike the eukaryotic (true) cells, bacteria do not have a
membrane enclosed nucleus.
• The nucleoid is a region of cytoplasm where the chromosomal
DNA is located.
• It is not a membrane bound nucleus, but simply an area of the
cytoplasm where the strands of DNA are found.
 Plasmids
• small extra-chromosomal DNA
• contain genes for antibiotic resistance or virulence.
• Structure Similar to most bacterial chromosomes, but considerably
smaller.
• plasmids are covalently closed circular DNA
• In a few species linear plasmids have been found.
• Size : Chromosomal DNA is typically about 4000 kb,
• plasmid DNA ranges from 1-200 kb.
• Number of plasmids: 1-700 copies of plasmid in a cell.
 Plasmid Function
• The function of plasmids is not always known, but they are not normally
essential for survival of host, although their presence
generally gives the host some advantage.
• Antibiotic resistance - Some plasmids code for proteins that degrade
antibiotics-a big advantage for pathogens.
• Some encode for proteins which confer virulence factors on the host.
For example- E. coli plasmid Ent P307 codes for an enterotoxin which
makes E. coli pathogenic.
• Conjugative plasmids - These allow exchange of DNA between bacterial
cells.
 Plasmids
• Plasmids and the associated traits can be transferred between bacteria,
even from one bacterial species to another.
• Plasmids are not involved in reproduction.
• Plasmids replicate independently of the chromosome.
• Plasmids are passed to other bacteria by two means.
• For most plasmid types, copies in the cytoplasm are passed on to
daughter cells during binary fission.
 Plasmids
• Other types of plasmids ,form tube like structure at the surface called a
pilus that passes copies of the plasmid to other bacteria during
conjugation, a process by which bacteria exchange genetic information.
• Plasmids have been shown to be instrumental in the transmission of
special properties, such as antibiotic drug resistance, resistance to heavy
metals, and virulence factors necessary for infection of animal or plant
hosts.
• The ability to insert specific genes into plasmids have made them
extremely useful tools in the area of genetic engineering/RDNA
Technology .
 Ribosomes- protein synthesis machinery
• Consists of RNA and protein
• Abundant in cytoplasm
• Often grouped in long chains called polyribosomes.
• give the cytoplasm of bacteria a granular appearance in EM.
• smaller than the ribosomes in eukaryotic cells-but have a similar
function
• Bacterial ribosomes have sedimentation rate of 70S; their subunits
have rates of 30S and 50S.
• The unit used to measure sedimentation velocity is Svedberg
 Ribosomes
• They translate the genetic code from the molecular language of nucleic
acid to that of amino acids—the building blocks of proteins.
• Bacterial ribosomes are similar to those of eukaryotes, but are smaller and have a slightly different
composition and molecular structure.
• Bacterial ribosomes are never bound to other organelles as they sometimes are bound to the
endoplasmic reticulum in eukaryotes, but are free-standing structures distributed throughout the
cytoplasm.
• There are sufficient differences between bacterial ribosomes and eukaryotic ribosomes that some
antibiotics will inhibit the functioning of bacterial ribosomes, but not a eukaryote's, thus killing
bacteria but not the eukaryotic organisms they are infecting.
• Streptomycin binds 70S ribosome and stops protein synthesis but it can not bind 80S ribosome of
eukaryotes and thereby eukaryotic cell remains unaffected.
 Bacterial Chromosome - Genophore
• The bacterial chromosome consists of a single, circle of
deoxyribonucleic acid.
• DNA is double stranded- two strands line up antiparrallel to
each other and the bases are linked together with hydrogen
bonds.
• It includes most of the genetic material of the organism .
 Bacterial Chromosome
• Unlike the DNA in eukaryotic cells, which resides in the nucleus,
DNA in bacterial cells is not sequestered in a membrane-bound
organelle but
appears as a long coil distributed through the cytoplasm.
• In many bacteria the DNA is present as a single, circular
chromosome and in some cases the DNA is linear rather than
circular.
• some bacteria may contain two chromosomes
 Bacterial Chromosome
• As in all organisms, bacterial DNA contains the four
nitrogenous bases adenine (A), cytosine (C), guanine (G), and
t
• The amount of DNA in bacterial chromosomes ranges from
580,000 base pairs in Mycoplasma gallinarum to 4,700,000
base pairs in E. coli to 9,140,000 base pairs in Myxococcus
xanthus.
 Inclusion bodies
• Inclusion bodies: Bacteria can have within their cytoplasm a variety of
small bodies collectively referred to as inclusion bodies.
• Some are called granules and other are called vesicles.
• Inclusions are considered to be nonliving components of the cell that do
not possess metabolic activity and are not bounded by membranes.
• The most common inclusions are glycogen, lipid droplets, crystals,
and pigments.
 Inclusion bodies - Granules
• Granules: Densely compacted substances without a membrane
covering.
• Nutrients and reserves may be stored in the cytoplasm in the form of
glycogen, lipids, polyphosphate, or in some cases, sulfur or nitrogen for
later use.
• Each granule contains specific substances, such as glycogen (glucose
polymer) and polyphosphate (phosphate polymer, supplies energy to
metabolic processes).
• Sulfur bacteria contains reserve granules of sulfur.
• These granules are depleted in starvation.
 Inclusion bodies-vesicles
• Some aquatic photosynthetic bacteria and cyano bacteria have rigid
gas-filled vacuoles and it helps in floating at a certain level - allowing
them to move up or down into water layers with different light
intensities and nutrient levels.
• Some magnetotactic bacterium, eg. Aquaspirillium magnetotacticum ,
stores Magnetitite (Ferric oxide). The presence of such magnetic
inclusions enables these bacteria to responds to magnetic fields.
 Microcompartments
• Microcompartments are widespread, membrane- bound
organelles that are made of a protein shell that surrounds and
encloses various enzymes.
• Carboxysomes are protein-enclosed bacterial microcompartments
that contain enzymes involved in carbon fixation.
• Magnetosomes are bacterial microcompartments, present in
magnetotactic bacteria, that contain magnetic crystals.
 Cell Envelope

• Plasma Membrane
• Periplasmic Space
• Cell Wall
• Outer membrane
 Plasma Membrane
• Phospholipid bilayer surrounding the cytoplasm and
regulates the flow of substances in and out of the cell.
• Consists of both lipids and proteins.
• Protects the cell from its surroundings.
• Selectively permeable to ions and organic molecules and
controls the movement of substances in and out.
• numerous proteins moving within or upon this layer are
primarily responsible for transport of ions, nutrients and waste
across the membrane.
 Periplasmic space
• Gram-negative bacteria : spacebetween the cytoplasmic
membrane and the cell wall and space found between cell
wall and the outer membrane
• Periplasm may constitute up to 40% of the total cell
volume in G-ve species.
• Gram-positive bacteria : space between the
cytoplasmic membrane and the cell wall.
• The periplasm is filled with water and proteins and is
reminiscent of the cytoplasm.
 Periplasmic Space
• However periplasm contains proteins and other
molecules distinct from those in the cytoplasm
because the membrane prevents the free exchange
between these two compartments.
• Periplasmic proteins have various functions in
cellular processes including: transport, degradation,
and motility.
• Periplasm controls molecular traffic entering and
leaving the cell.
[Link]
PERI PLASMIC SPACES
 Cell wall
• Outer covering of most cells that protects the bacterial cell
and gives it shape (spherical, rod and spiral).
• Composed of peptidoglycan (polysaccharides + protein)
• Mycoplasma are bacteria that have no cell wall and
therefore have no definite shape.

[Link]

CELL WALL
 Cell wall
• Peptidoglycan - molecule found only in bacterial cell walls.
• The rigid structure of peptidoglycan gives the bacterial cell shape,
surrounds the plasma membrane and provides prokaryotes with
protection from the environment
• From the peptidoglycan inwards all bacterial cells are similar.
• Going further out, the bacterial world divides into two major
classes: Gram-positive and Gram-negative .
• Amount and location of peptidoglycan in the cell wall
determines whether a bacterium is G+ve or G-ve
Peptidoglycan = (polysaccharides + protein),
• Peptidoglycan (murein) - huge polymer of interlocking chains
composed of similar monomers.
• peptidoglycan is made from polysaccharide chains cross-
linked by peptides containing D-amino acids
• The backbone of the peptidoglycan molecule is
composed of two derivatives of glucose:
• N-acetylglucosamine (NAG)
• N-acetlymuramic acid (NAM).
• The NAG and NAM strands are connected by inter peptide
bridges.
 Gram-positive Cells

• G+ve bacteria possess thick cell wall containing many

layers of peptidoglycan and teichoic acids.
• In G+ ve cells, peptidoglycan is the outermost structure
and makes up as much as 90% of the thick compact
cell wall.

[Link]

GRAM POSITIVE VS GRAM NEGATIVE

 Gram-negative

• G-ve bacteria have relatively thin cell wall consisting of

few layers of peptidoglycan surrounded by a second lipid
membrane containing lipopolysaccharides and
lipoproteins
• Peptidoglycan makes up only 5 – 20% of the cell wall
and is not the outermost layer, but lies between the
plasma membrane and an outer membrane.
 Gram Staining
• Developed in 1884 by Danish scientist Christian Gram.
• It is a differential stain.
• In this, bacteria are first stained with crystal violet, then
treated with a mordant - a solution that fixes the stain
inside the cell.
• Bacteria are then washed with a decolorizing agent, such as
alcohol, and counterstained with safranin, a light red dye.
[Link]

Gram Staining
 Gram Staining
• Gram-positive bacteria are those that are stained dark
blue or violet by Gram staining.
• Gram-negative bacteria cannot retain the crystal violet
stain, instead take up the counterstain and appearred or
pink.
• The walls of gram-positive bacteria have more
peptidoglycans than do gram-negative bacteria. Thus,
gram-positive bacteria retain the original violet dye and
cannot be counterstained.
 Cell wall
• If the bacterial cell wall is entirely removed, it is called a
protoplast while if it's partially removed, it is called a
spheroplast.
• Antibiotics such as penicillin inhibit the formation of
peptidoglycan cross-links in the bacterial cell wall.
• The enzyme lysozyme, found in human tears, also digests the
cell wall of bacteria and is the body's main defense against
eye infections. [Link]

CELL WALL
 outer membrane
[Link]

• Similar to the plasma membrane, but is less permeable .

• This membrane has tiny holes or openings called porins.
• Porins block the entrance of harmful chemicals and antibiotics, making G-ve bacteria
much more difficult to treat than G+ve cells.
• Composed of lipopolysaccharides
[Link]
(LPS).
• LPS is a harmful substance classified as an endotoxin.
• Lipopolysaccharides, which acts as an endotoxin, are composed of polysaccharides
and lipid A (responsible for much of the toxicity of G-ve bacteria).
• These differences in structure can produce differences in antibiotic susceptibility
• Ex: vancomycin can kill only Gram +ve bacteria and is ineffective against Gram -ve
pathogens, such as Haemophilus influenzae or Pseudomonas aeruginosa.
 External structures
……………………..
• Flagella …………………………………...
[Link]

• Pili/fimbriae
• Capsule/slime layer
 Flagella
• Singular: flagellum
• Long, whip-like semi-rigid cylindrical structures that aids in
cellular locomotion
• Function much like the propeller on a ship.
• about 20 nm in diameter and up to 20 micromts in length.
• Diameter of a prokaryotic flagellum is about 1/10 th of that of
eukaryotic.
• Flagella are driven by the energy released by the transfer of
ions down an electrochemical gradient across the cell
membrane.
 Flagella
• Made up of protein subunits called flagellin.
• Each flagellum is attached to cell membrane with the
help of proteins other than flagellin.
• The basal region has a hook like structure and a
complex basal body. The basal body consists of a central
rod or shaft surrounded by a set of rings.
 Flagella
• Bacterial spp differ in the number and arrangement of
flagella on their surface.
• Bacteria may have one, a few, or many flagella in different
positions on the cell.
• Monotrichous - single flagellum
• amphitrichous a flagellum at each end lophotrichous -
clusters of flagella at the poles of the cell
• peritrichous - flagella distributed over the entire surface
of the cell .
Arrangement of flagella
 Flagella
• Motile bacteria are attracted or repelled by certain stimuli in
behaviors called taxis: these include chemotaxis, phototaxis,
and magnetotaxis.
• The flagella beat in a propeller-like motion to help the
bacterium move toward nutrients; away from toxic chemicals;
towards the light (photosynthetic cyanobacteria).
• Prokaryotes exhibit a variety of movements:
move , swim ,tumble ,glide, swarm in response to
environmental stimuli.
 FIMBRIAE AND PILI
• Hollow, hair like structures made of protein
• Involved in attachment to solid surfaces or to other cells and
are essential for the virulence of some bacterial pathogens.
• Fimbriae fine filaments of protein just 2–10 nm in diameter
and up to several micrometers in length.
• They are distributed over the surface of the cell, and
resemble fine hairs when seen under the electron
microscope.
[Link]
 FIMBRIAE AND PILI

• Pili: (sing. pilus) are cellular appendages, slightly larger than

fimbriae
• Involved in attachment to surfaces.
• Specialized pili, the sex pili, allows the transfer of genetic
material from one bacteria to another in a process called
conjugation where they are called conjugation pili or "sex pili".
• type IV pili - generate movement.
• Helps in colonization and pathogenicity.
 Glycocalyx
• Glycocalyx : sticky coating produced by many bacteria
covering the surface of cell.
• The glycocalyx is composed of polysaccharides (sugars)
and proteins.
• The bacterial glycocalyx has 2 forms
• a highly structured rigid capsule
• a disorganised loose slime layer -
• Capsules are found on many pathogenic bacteria
 Glycocalyx

• The glycocalyx has several functions including : protection,

attachment to surfaces and formation of biofilms.
• The glycocalyx helps protect the bacteria cell by preventing
immune cells from attaching to it and destroying it through
phagocytosis.

[Link]
 Bacterial reproduction
• Cell growth and reproduction by cell division are tightly linked in
unicellular organisms.
• Bacteria grow to a fixed size and then reproduce through binary
fission, a form of asexual reproduction
• Under optimal conditions, bacteria can grow and divide extremely
rapidly, and bacterial populations can double as quickly as every 9.8
minutes.
• In cell division, two identical clone daughter cells are produced.
• Budding involves a cell forming a protrusion that breaks away and
produces a daughter cell [Link]
 Binary fission
• Most prokaryotes reproduce by a process of binary fission, in which the cell grows in
volume until it divides in half to yield two identical daughter cells.
• Each daughter cell can continue to grow at the same rate as its parent.
• For this process to occur, the cell must grow over its entire surface until the time of cell
division, when a new hemispherical pole forms at the division septum in the middle of
the cell.
• The septum grows inward from the plasma membrane along the midpoint and forms
as the side wall which pinches inward, dividing the cell in two.
• In order for the cell to divide in half, the peptidoglycan structure must be different
in the hemispherical cap than in the straight portion of the cell wall, and different wall-
cross-linking enzymes must be active at the septum than elsewhere.
[Link]
 Binary fission

• Binary fission begins with the single DNA molecule replicating and
both copies attaching to the cell membrane.
• Next, the cell membrane begins to grow between the two DNA
molecules. Once the bacterium just about doubles its original size, the
cell membrane begins to pinch inward.
• A cell wall then forms between the two DNA molecules dividing the
original cell into two identical daughter cells

Microbiology - Bacteria Growth, Reproduction, Classification

[Link]
 Budding
• A group of environmental bacteria reproduces by budding.
• In this process a small bud forms at one end of the mother cell
• As growth proceeds, the size of the mother cell remains about constant, but the
bud enlarges.
• When the bud is about the same size as the mother cell, it separates. This type
of reproduction is analogous to that in budding fungi, such as brewer’s yeast
(Saccharomyces cerevisiae).
• One difference between fission and budding is that, in the latter, the mother cell
often has different properties from the offspring.
• Ex: In some strains, mother cells have a flagellum and are motile, whereas the
daughter buds lack flagella.
Budding of Yeast Cells
[Link]
BACTERIAL RECOMBINATION
 Three mechanisms of genetic recombination

• Conjugation
• Transformation
• Transduction
genetic recombination strategies of bacteria CONJUGATION, TRANSDUCTION AND TRANSFORMATION
[Link]

Transformation, Conjugation, Transposition and Transduction

[Link]
 CONJUGATION

• Two bacterial cells come together and mate such that a gene transfer occurs
between them.
• Can only occur between cells of opposite mating types.
– The donor (or "male") carries a fertility factor (F+).
– The recipient ("female") does not (F−).
• One cell, the donor cell (F+), gives up DNA; and another cell, the recipient cell (F−), receives the DNA.
• The transfer is nonreciprocal, and a special pilus called the sex pilus joins the donor and recipient
during the transfer.
• The channel for transfer is usually a special conjugation tube formed during
contact between the two cells.
• The DNA most often transferred is a copy of the F factor plasmid.
• The factor moves to the recipient, and when it enters the recipient, it is copied to produce a
double-stranded DNA for integration.
 BACTERIAL TRANSFORMATION

• Discovered by Frederick Griffith in 1928.

• Many bacteria can acquire new genes by taking up DNA molecules (ex:
plasmid) from their surroundings.
• When bacteria undergo lysis, they release considerable amounts of DNA
into the environment.
• This DNA may be picked up by a competent cell- one capable of taking
up the DNA and undergoing a transformation.
• To be competent, bacteria must be in the logarithmic stage of growth, and a
competence factor needed for the transformation must be present.
 BACTERIAL TRANSDUCTION
• Bacterial viruses ( bacteriophages) transfer DNA fragments
from one bacterium (the donor) to another bacterium (the
recipient).
• The viruses involved contain a strand of DNA enclosed in an
outer coat of protein.
 After a bacteriophage enters a bacterium, it may encourage the bacterium to make
copies of the phage.
 At the conclusion of the process, the host bacterium undergoes lysis and releases new
phages. This cycle is called the lytic cycle.
Under other circumstances, the virus may attach to the bacterial chromosome and
integrate its DNA into the bacterial DNA. It may
remain here for a period of time before detaching and continuing its replicative process.
This cycle is known as the lysogenic cycle.
Under these conditions, the virus does not destroy the host bacterium, but remains in a
lysogenic condition with it. The virus is called a temperate phage, also known as a
prophage.
 At a later time, the virus can detach, and the lytic cycle will ensue.
 It will express not only its genes, but also the genes acquired from the donor
bacterium.
GROWTH OF BACTERIA
 Bacterial Growth
• Growth of Bacteria is the orderly increase of all the
chemical constituents of the bacteria.
• Multiplication is the consequence of growth.
• Death of bacteria is the irreversible loss of ability to
reproduce.

Microbiology - Bacteria Growth, Reproduction, Classification

[Link]
 Generation /doubling time
• Generation time (g) : The time it takes the cells to double.
• The average generative time is about 20-30 minutes in majority of
medically important bacteria.
• They are some exceptions among pathogenic bacteria.
• Mycobacterium tuberculosis - 18 hrs.
• Mycobacterium leprae -10-20 days
• Length of generative time is in direct dependence on the length of
incubation period of infections.
 Growth Kinetics
• Bacterial growth follows four phases.
• lag phase
• log phase
• stationary phase
• death phase
Bacterial Growth Curve
[Link]
 Lag phase
• Immediately following the seeding of a culture medium.
• A period of adaptation for the cells to their new environment
• cells are adapting to the high-nutrient environment and preparing for fast
growth.
• The lag phase has high biosynthesis rates, as proteins and metabolic
intermediates are built up in adequate quantities for rapid growth &
multiplication to proceed.
• New enzymes are synthesized.
• A slight increase in cell mass and volume, but no increase in cell number.
 Duration of the lag phase varies with

- the species
- size of inoculum - Prolonged by low inoculum volume, poor inoculum
condition (high % of dead cells)
- age of inoculum
- Nature of the culture medium (Prolonged by nutrient- poor medium)
- And environmental factors like temperature, pH etc
 Log/Exponential growth phase

• In this phase, the cells have adjusted to their new environment and multiply
rapidly (exponentially)
• The bacteria will grow and divide at a doubling time characteristic of the
strains and determined by the conditions during the exponential phase.
• During this phase, the number of bacteria will increase to 2n, in which n is
the [Link] generations.
• Balanced growth –all components of a cell grow at the same rate.
Deceleration growth phase

Very short phase, during which growth decelerates due to either:

• Depletion of one or more essential nutrients
• The accumulation of toxic by-products of growth (e.g. Ethanol in yeast
fermentations)
• Period of unbalanced growth: Cells undergo internal restructuring to
increase their chances of survival
 Stationary Phase

With the exhaustion of nutrients or build-up of toxic waste substances and

secondary metabolic products in the medium , the bacteria stop growing and
enter the stationary phase.
- The growth rate equals the death rate – The number of progeny cells
formed is just enough to replace the number of cells that die.
- There is no net growth in the organism population – The viable count remains stationary as
an equilibrium exists between the dying cells and newly formed cells.
 Death Phase

- Phase of decline
- The living organism population decreases with time, due to a lack of
nutrients and accumulation of toxic metabolic by-products.
- Cell death may also be caused by autolytic enzymes.
 Generation times

Bacterium Medium Generation Time (minutes)

Escherichia coli Glucose-salts 17
Bacillus megaterium Sucrose-salts 25
Streptococcus lactis Milk 26
Streptococcus lactis Lactose broth 48
Staphylococcus aureus Heart infusion broth 27-30

Lactobacillus acidophilus Milk 66-87

Rhizobium japonicum Mannitol-salts-yeast extract 344-461

Mycobacterium tuberculosis Synthetic 792-932

Treponema pallidum Rabbit testes 1980
Factors Required for Bacterial Growth

The requirements for bacterial growth are:

(A) Environmental factors

(B) Sources of metabolic energy.
 Environmental Factors
Affecting Bacterial Growth
microbiology factor affecting bacterial growth
[Link]
 Nutrients
• Nutrients in growth media must contain all the elements
necessary for the synthesis of new organisms.
• Hydrogen donors and acceptors
• Carbon source
• Nitrogen source
• Minerals : sulphur and phosphorus
• Growth factors: amino acids, purines, pyrimidines; vitamins
• Trace elements: Mg, Fe, Mn.
 Temperature
• Microorganisms are sensitive to temperature changes

– Usually unicellular
– Enzymes have temperature optima
– If temperature is too high, proteins denature, including enzymes, carriers
and structural components

• Temperature ranges are enormous (-20 to 100oC)

Temperature
– Organisms exhibit distinct cardinal temperatures (minimal,
maximal, and optimal growth temps)
– If an organism has a limited growth temperature range =
stenothermal (e.g. N. gonorrhoeae)
– If an organism has a wide growth temperature range =
eurythermal (E. faecalis)
Temperature
Psychrophiles can grow well at 0oC, have optimal
growth at 15oC or lower, and usually will not grow
above 20oC
• Arctic/Antarctic ocean
• Protein synthesis, enzymatic activity and transport systems
have evolved to function at low temperatures
• Cell walls contain high levels of unsaturated fatty acids (semi-fluid
when cold)
Temperature
– Psychrotrophs can also grow at 0oC, but have growth optima between
20oC and 30oC, and growth maxima at about 35oC

• Many are responsible for food spoilage in refrigerators

– Mesophiles have growth minima of 15 to 20oC, optima of 20 to 45oC, and

maxima of about 45oC or lower
• Majority of human pathogens
Temperature
–Thermophiles have growth minima around 45oC, and
optima of 55 to 65oC
• Hot springs, hot water pipes, compost heaps
• Lipids in PM more saturated than mesophiles.
–Hyperthermophiles have growth minima around 55oC
and optima of 80 to 110oC
• Sea floor, sulfur vents
Effect of temperature
Temperature optima of bacteria
 pH
– pH is the negative logarithm of the hydrogen ion
concentration

– Acidophiles grow best between pH 0 and 5.5

– Neutrophiles grow best between pH 5.5 and 8.0
– Alkalophiles grow best between pH 8.5 and 11.5
– Extreme alkalophiles grow best at pH 10.0 or higher
 pH
– Sudden pH changes can inactivate enzymes and damage
plasma membrane
• Reason for buffering culture medium, usually with a weak
acid/conjugate base pair (e.g. KH2PO4/K2HPO4 – monobasic
potassium/dibasic potassium)
Bacterial growth at various pH
 pH profiles for some prokaryotes
Organism Minimum pH Optimum pH Maximum pH
Thiobacillus thiooxidans 0.5 2.0-2.8 4.0-6.0
Sulfolobus acidocaldarius 1.0 2.0-3.0 5.0
Bacillus acidocaldarius 2.0 4.0 6.0
Zymomonas lindneri 3.5 5.5-6.0 7.5
Lactobacillus acidophilus 4.0-4.6 5.8-6.6 6.8
Staphylococcus aureus 4.2 7.0-7.5 9.3
Escherichia coli 4.4 6.0-7.0 9.0
Clostridium sporogenes 5.0-5.8 6.0-7.6 8.5-9.0
Erwinia caratovora 5.6 7.1 9.3
Pseudomonas aeruginosa 5.6 6.6-7.0 8.0
Thiobacillus novellus 5.7 7.0 9.0
Streptococcus pneumoniae 6.5 7.8 8.3
Nitrobacter sp 6.6 7.6-8.6 10.0
 Oxygen concentration

– Obligate aerobes are completely dependent on

atmospheric O2 for growth
• Oxygen is used as the terminal electron acceptor for electron
transport in aerobic respiration
– Facultative anaerobes do not require O2 for growth, but do
grow better in its presence
– Aerotolerant anaerobes ignore O2 and grow equally well
whether it is present or not
 Oxygen concentration

• –Obligate (strict) anaerobes do not tolerate O2 and die in its

presence.
• –Microaerophiles are damaged by the normal atmospheric level of
O2 (20%) but require lower levels (2 to 10%) for
growth
 Oxygen and growth
Environment

Group Aerobic Anaerobic O2 Effect

Obligate Aerobe Growth No growth Required (utilized for aerobic

respiration)
Growth if Required but at levels below 0.2
Microaerophile No growth
level not too atm
high
Obligate Anaerobe No growth Growth Toxic

Not required for growth but utilized

Facultative Growth Growth
when available
(An)aerobe

Aerotolerant Growth Growth Not required and not utilized

Anaerobe
Anaerobic growth chambers
 Water availability
• Water is solvent for biomolecules, and its availability is critical for cellular
growth
• The availability of water depends upon its presence in the atmosphere
(relative humidity) or its presence in solution or a substance (water
activity, (Aw))
• Aw of pure water (100%) is 1.0; affected by dissolved solutes such as
salts or sugars.
• Microorganisms live over a range of aW from 1.0 to 0.7. The aW of human
blood is 0.99; seawater = 0.98; maple syrup = 0.90; Great Salt Lake = 0.75.
Water activities in agricultural soils range between 0.9 and 1.0.
Moisture Content and Water Activity
[Link]
 Effect of salt on growth

Environmental Influences on Microbial Growth-Salt Tolerance & pH testing

[Link]
 Pressure
–Barotolerant organismsare adversely

• affected by increased pressure, but not as severely as are nontolerant

organisms
• –Barophilic organisms require, or grow more rapidly in the presence of
increased pressure
•–Light: Optimum condition for growth is darkness.
 Radiation

-Ultraviolet radiation damages cells by causing the formation of

thymine dimers in DNA.
– Ionizing radiation such as X rays or gamma rays are even more
harmful to microorganisms than ultraviolet radiation
• Low levels produce mutations and may indirectly result in
death
• High levels are directly lethal by direct damage to
cellular macromolecules or through the production of oxygen
free radicals
 (B) Sources of Metabolic Energy

• Mainly three mechanisms generate metabolic energy.

These are
• Fermentation
• Respiration and
• Photosynthesis.
An organism to grow, at least one of these mechanisms
must be used.
Continuous Culture
Techniques
Used to maintain cells in the exponential growth
phase at a constant biomass for Concentration
extended periods of time
• Conditions are met by continual provision of
nutrients and removal of wastes = OPEN SYSTEM
• Constant conditions are maintained
• Balanced and Unbalanced
Growth
• Balanced (exponential) growth occurs when all cellular
components are synthesized at constant rates relative to one
another
• Unbalanced growth occurs when the rate of synthesis of
some components change relative to the rate of synthesis of
other components.
– This usually occurs when the environmental conditions
change
 MORPHOLOGICAL
CHARACTERISTICS OF
BACTERIA
SIZE- SHAPE-ARRANGEMENT
Microbiology of Bacterial Morphology & Shape
[Link]
 MORPHOLOGY
• Bacteria display a wide diversity of shapes and sizes called
morphologies
• Cannot be seen with human eyes (microscopic)
• Their presence was only first recognized in 1677, when the Dutch naturalist Antonie van
Leeuwenhoek saw microscopic organisms in a variety of substances with the aid of
primitive microscopes.
• Now bacteria are usually examined under light microscopes capable of more
than 1,000-fold magnification
• Details of their internal structure can be observed only with the aid of much more
powerful transmission electron microscopes.
• Unless special phase-contrast microscopes are used, bacteria have to be stained
with a coloured dye so that they will stand out from their background.
 Size
• Bacteria are the smallest living creatures
• Most bacteria are 0.2 um in diameter and 2-8 um in length.
• Bacterial cells are about one tenth the size of eukaryotic cells
• are typically 0.5 – 5.0 micrometres in length.
• Giant bacteria for example, Thiomargarita namibiensis, Titanospirillum
namibiensis and Epulopiscium fishelsoni — are up to half a mm long and are visible
to the unaided eye
• E. fishelsoni reaches 0.7 mm.
• Among the smallest bacteria are members of the genus Mycoplasma,
which measure about 0.1 to 0.25 μm in diameter, as small as the largest
viruses.
• Some bacteria may be even smaller, but these ultramicro bacteria are not
well-studied.
 Size
• E. coli, a normal inhabitant of the intestinal tract of humans and animals, is
about 2 μm long and 0.5 μm in diameter
• spherical cells of Staphylococcus aureus - up to 1 μm in diameter.
• the rod-shaped Bordetella pertussis, causative agent of whooping cough - 0.2 to
0.5 μm in diameter and 0.5 to 1 μm in length
• corkscrew-shaped Treponema pallidum, causative agent of syphilis averaging only
0.15 μm in diameter but 10 to 13 μm in length.
• Some bacteria are relatively large, such as Azotobacter, which has diameters of 2
to 5 μm or more
• cyanobacterium Synechococcus averages 6 μm by 12 μm
• Achromatium, which has a minimum width of 5 μm and a maximum length
of 100 μm, depending on the species.
 Cell Shape
• Bacteria come in a wide variety of shapes.
• Coccus – are spherical or oval cells.
• Bacillus - are round-ended cylinder shaped cells.
• Vibrios comma shaped ,curved rods and derive the name from
their characteristic vibratory motility
• Spirilla – are rigid spiral forms(coil).
• Spirochetes - are long, slender, and flexible spiral forms(from speira
meaning coil and chaite meaning hair)
• Filamentous – resembles radiating rays of sun
 Cell Shape
• coccobacilli - Some bacilli are so short and fat that they look like
cocci and are referred to as coccobacilli.
• A small number of species even have tetrahedral
or cuboidal shapes.
• More recently, bacteria were discovered deep under the Earth's
crust that grow as long rods with a star-shaped cross-section.
• The large surface area to volume ratio of this morphology may give
these bacteria an advantage in nutrient-poor environments.
 Cell Shape
• is generally characteristic of a given bacterial species
• but can vary depending on growth conditions.
• Some bacteria have complex life cycles involving the production of stalks
and appendages (e.g. Caulobacter) and some produce elaborate
structures bearing reproductive spores (e.g. Myxococcus, Streptomyces).
• Bacteria generally form distinctive cell morphologies when examined
by light microscopy and distinct colony morphologies when grown on
Petri plates.
• These are often the first characteristics observed by a microbiologist to
determine the identity of an unknown bacterial culture
 Cell Shape
• This wide variety of shapes is determined by the bacterial cell
wall and cytoskeleton
• Shape of the cell is important because it can influence the
ability of bacteria to acquire nutrients, attach to surfaces,
swim through liquids and escape predators
 Arrangement of cells
• Cellular arrangements occur singularly, in pairs, in chains and in
clusters.
 Bacilli
• Diplobacilli (2 cells), tetrad (4 cells), palisade (two cells
arranged parallel) or sterptobacilli (chain arrangement)e.g
[Link] and Salmonella.
Cocci
Bacilli
Other shapes
 CULTURAL
CHARACTERISTICS OF
BACTERIA
Cultural characteristics & Colony morphology of Bacterial colony
[Link]
• Culture techniques are designed to promote the growth and
identify particular bacteria,while restricting the growth of the other
bacteria in the sample.
• In the laboratory, bacteria are usually grown using solid growth
media such as agar plates or liquid media such as broth.
• Solid, agar-based media can be used to identify colonial characteristics
(shape, size, elevation, margin type) and to isolate pure cultures of a
bacterial strain
• liquid growth media are used when measurement of growth or large
volumes of cells are required.
• Growth in stirred liquid media occurs as an even cell suspension,
making the cultures easy to divide and transfer
• isolating single bacteria from liquid media is difficult.
• The use of selective media (media with specific nutrients added or
deficient, or with antibiotics added) can help identify specific org’s.
• Most laboratory techniques for growing bacteria use high levels of
nutrients to produce large amounts of cells cheaply and quickly.
• However, in natural environments nutrients are limited, meaning that
bacteria cannot continue to reproduce indefinitely
 Cultural characteristics
Basic conditions for cultivation
• Optimum environmental moisture. It is possible to cultivate bacteria in
liquid media or solid media with a gelling agent (agar) binding about 90%
of water.
• Optimum temperature for cultivation of bacteria of medical importance
is about 370C. Saprophytic bacteria are able to grow at lower temperatures.
• Optimum pH of culture media is usually 7.2-7.4 Lactobacillus sppneed acid
pH and vibrio cholera needs alkaline pH for the growth.
• Optimum constituents of bacteriological culture media.
• All culture media share a number of common constituents necessary to
enable bacteria to grow in vitro.
 Optimum Quantity of oxygen in
cultivation environment.
• Bacteria obtain energy either by oxidation or by fermentation i.e., oxidation – reduction
procedure without oxygen.
• Bacteria are classified into four basic groups according to their relation to
atmospheric oxygen:
• Obligate aerobes: Reproduce only in the presence of oxygen
• Facultative anaerobes : reproduce in both aerobic and anaerobic environments. Their
complete enzymatic equipment allows them to live and grow in the presence or
absence of oxygen.
• Obligate anaerobes: grow only in the absence of free oxygen (i,e unable to grow and
reproduce in the presence of oxygen). Some species are so sensitive that they die if
exposed to oxygen.
• Anaerobic Aerotolerant: microbes do not need oxygen for their growth and it is not
fatal for them
 Colony morphology
• Form - the basic shape of the colony ex: circular,
filamentous etc.
• Size – The diameter of the colony.
• Elevation - This describes the side view of a colony. Turn the Petri dish
on end.
• Margin/border - magnified shape of the edge of the colony
• Surface - colony appearance
ex: smooth, glistening, rough, wrinkled or dull.
• Opacity - ex transparent (clear), opaque, translucent (like looking
through frosted glass), etc.
• Colour (pigmentation) ex: white, buff, red, purple, etc.
 Colony morphology
• Colony morphology is a method that scientists use to describe the
characteristics of an individual colony of bacteria growing on agar in
a Petri dish. It can be used to help to identify them.
• Each distinct colony represents an individual bacterial cell or group
that has divided repeatedly. Being kept in one place, the resulting
cells have accumulated to form a visible patch.
• Most bacterial colonies appear white or a creamy yellow in colour,
and are fairly circular in shape.
 Effect of media
• different types of media, which contain different nutrients can
affect the cultural characteristics of bacteria.
• Some types of media are much more nutritive and will encourage hearty
growth. Some types of media may restrict growth.
• Colonial morphology may also be affected by the temperature at which
the bacteria is incubated. Some bacteria grow better at body
temperature and grow weakly at room temperature, or vice versa.
• Some bacteria express certain characteristics, such as the formation of
pigment, more strongly at some temperatures than at others.
CLASSIFICATION
OF BACTERIA
 Bacterial Classification Based on Shapes
• Bacilli: Rod shaped bacteria.
• Diplobacilli, tetrad , palisade (two cells arranged parallel) or sterptobacilli
(chain arrangement). e.g. [Link] and Salmonella

• Coccus: Spherical or oval cells shaped bacteria which is further classified as

monococcus, diplococci, streptococci ,Staphylcocci
e.g. Staphylococcus and Streptococcus

• Spiral: Spiral shaped bacteria are called spirilla

e.g. Treponema and Borellia
sub divided into spirilla (rigid spiral forms) and
spirochetes(flexible spiral forms).
• Comma shaped: Vibrio
• Branching filamentous forms : Actinomycetes
 Bacterial Classification Based on Staining
Methods
• Gram positive bacteria - take up crystal violet dye and retain their
blue or violet color.
Gram negative bacteria - do not take up crystal violet dye, and
thus appear red or pink.
 Classification Based on Respiration
• Aerobic Respiration : sugars are broken down in the presence of
oxygen to produce carbon dioxide, water, and energy.
• Anaerobic Respiration : anaerobic respiration breaks down
sugars and releases energy in the absence of oxygen.
• anaerobic respiration is typically slower and less efficient than
aerobic respiration.
• anaerobic respiration involves chemicals other than oxygen and
carbondioxide.
 Classification Based on Respiration
• Facultative Anaerobic Respiration : Facultative Anaerobes are
able to perform either aerobic / anaerobic respiration
depending on the oxygen content of their environment.
• Ex: Coliform bacteria
• Microaerophiles : sugars are broken down in the presence of
minute amounts of oxygen to produce energy.
 Classification Based on Environment
• Mesophiles - which require moderate temp to survive. Neutrophiles -
require moderate conditions to survive. Extremophiles - can survive in
extreme conditions. Acidophiles - which can tolerate low pH conditions.
Alkaliphiles - which can tolerate high pH conditions. Thermophiles -
which can resist high temperature. Psychrophiles - can survive extremely
cold conditions. Halophiles - can survive in highly saline conditions.
Osmophiles - can survive in high sugar osmotic conditions.
 Classification Based on Flagella
• Atrichous (no flagella),
• monotrichous (uni flagella)
• amphitrichous (bi flagella)
• polytrichous (more flagella)
 Classification Based on Spore Formation

• spore forming
• non-spore forming
 Classification Based on their association with host

• Beneficial
• Pathogenic
• Harmless
 Nutritional Source
• bacteria are also classified based on the type of energy source
utilized by them for survival.
• Autotrophs: obtain the carbon it requires from carbon dioxide
• Photoautotrophs: directly use sunlight in order to produce sugar
from carbon dioxide.
• Chemoautotrophs : depend on various chemical reactions.
• use inorganic energy sources, such as hydrogen sulfide,
elemental sulfur, ferrous iron, molecular hydrogen, and
ammonia.
 Nutritional Source
• Heterotrophs : Heterotrophic bacteria obtain sugar from the
environment they are in (ex: the living cells or organisms they
are in).
• symbiotic
• saprophytes
• parasite
 Capsule
• Capsulated
• Encapsulated
 Classification of bacteria
• With over millions of bacteria present in the planet, it is not an easy job
to identify, isolate and study a particular species or particular bacteria as
such.
• Microbiologists categorized bacteria based on basic and important
factors making all the bacteria fall under any one of the categories and
thus making the process of isolation and identification much easier.
• Bacteria are classified based on various factors like shape (morphology),
Cell wall structure, Respiration (metabolism), type of nutritional source,
characteristic and environmental factor.
 Bacteria are classified based on various
factors

• shape (morphology)
• Cell wall structure
• Respiration (metabolism)
• type of nutritional source
• characteristic
• environmental factor etc.
• Chemostat
– A continuous culture device that maintains a constant growth
rate by:
• supplying a medium containing a limited amount of an essential nutrient at
a fixed rate
• removing medium that contains microorganisms at the same rate
– As fresh media is added to the chamber, bacteria are removed
– Limiting nutrients control growth rates
– Cell density depends on nutrient concentration
• Turbidostat

A continuous culture device that regulates the flow rate of media through the
vessel in order to maintain a predetermined turbidity or cell density

• There is no limiting nutrient

• Absorbance is measured by a photocell (optical sensing device)
REFERENCES
THANK YOU

For queries
Email: tejinder.e7881@[Link]