BGB- 201- Introduction to

Microbiology
Unit-1
History of microbiology
Prokaryotic & Eukaryotic cell
Ultra structure of bacteria
Gram positive and Gram negative bacteria
Cyanobacteria
Archaea
Mycoplasma
PPLO
 Introduction to Microbiology
• Defined as the study of organisms and agents too small (<1mm in
diameter) to be seen clearly by the naked eye [exception :
eukaryotic microbes like bread moulds, filamentous algae ]
• Microorganisms are diverse
• Classification according to five kingdom system-
Monera- ( Bacteria, cyanobacteria – blue green algae)
Protista - (unicellular algae- chlamydomonous, protozoa)
Fungi – (Fungi)
Animalia- (Porifera, coelentrata, Platyhelminthes, aschihelminthes,
annelida, arthropoda, mollusca, Echinodermata, Chordata)
Plantae- multicellular algae, bryophytes, pteridophytes,
gymnosperms, angiosperms
[Microorganisms except viruses were placed in first three kingdoms]
rRNA comparision studies led to the division of
organisms into three domains-
Bacteria (true bacteria, eubacteria)
• single celled prokaryotes
• No nucleus or membrane bound organelles
• Genetic information is stored in a single loop of DNA
• Cell wall contain structural molecule peptidoglycan
• Abundant in soil, water, air, major inhabitants of skin,
mouth, and intestines. Beneficial roles in biosphere
(Cycling dead plants and animals), cyanobacteria
(produce significant amount of oxygen by
photosynthesis)
• They divide by Binary Fission (into two equal halves)
 Structure : Bacteria

A plasmid is a small DNA molecule within a cell that is physically

separated from a chromosomal DNA and can replicate independently.
They are most commonly found in bacteria as small circular, double-
stranded DNA molecules
Archaea-

• Prokaryotes but different from bacteria (Unique rRNA

sequences)
• Cell wall lack peptidoglycan
• Live in extreme environment and divided into three groups-
Methanogens- produce methane as waste product from
respiration
Halophiles (halo-salty, philes-loving) like those in Great salt lake
and Dead sea
Thermophiles –(Therma- heat, philes- loving)- live in hot
sulfurous water, such as hot springs at Yellowstone National
Park.
• Non pathogenic
Eucarya

• Include microorganisms that are classified as

Protists and fungi
• Include animals and plants
Protists-
larger than procaryotes (include unicellular algae like
chlamydomonas, protozoa, slime molds, and water molds)
Algae
• photosynthetic eukaryotes and together with the
cyanobacteria produce about 75% of the planet’s oxygen.
[As photosynthesizers, algae need light, water, and carbon dioxide
for food production and growth]
• Cell wall is made of cellulose
• abundant in freshwater and salt water, in soil, and in association
• with plants (Lichen)
• foundation of aquatic food chains.
Protozoa
• Unicellular eukaryote, animal-like i.e. motile. Have
pseudopods (false feet)- these are extentions of the
cytoplasm
• Long appendages- flagella, Filamentous- cillia
• For nutrition- ingest organic matter and other microbes
• found in many different environments - some are
inhabitants of the intestinal tracts of animals, where they
aid in digestion of complex materials and synthesis of
vitamins( B and K) , Photosynthetic protozoa(Euglena)
• Few cause disease in humans and other animals .
• Reproduce by sexual and Asexual reproduction
• Sexual reproduction : male and female gamete fuse to
form zygote→ embryo→offspring (inherit the features
of both parents)
• Asexual reproduction : single organism produces
genetically identical offspring
Benefits:
rapid population growth
One organism can make whole population
Less likely to become extinct
Water molds

• found in the surface water of freshwater sources and

moist soil.
• feed on decaying vegetation- logs and mulch
Fungi

• Range from unicellular forms (yeasts) to multicellular

( molds and mushrooms)

• Molds and mushrooms form thin threadlike structures

called hyphae. They absorb nutrients from their
environment, including the organic molecules (as a source
of carbon and energy)
• Because of their metabolic capabilities, many fungi play
beneficial roles-(making bread rise, producing antibiotics,
and decomposing dead organisms)
• Other fungi cause diseases in plants, human and animals
Viruses

• Smallest of all microbes (10,000 times smaller than a typical

bacterium)
• Acellular
• Need host cell machinery in order to replicate
• Cause many animal and plant diseases - smallpox, rabies,
influenza, AIDS, the common cold, and some cancers.
• They have their own classification system
 History
• Earlier there was a suggestion that disease was caused by some
invisible living creatures
• the first person to publish accurate observations of
microorganisms was the Antony van Leeuwenhoek (1632–1723)
of Delft, The Netherlands
• Microscopic observations were not sufficient enough so
techniques of isolating and culturing of microorganisms in lab
came into picture
• Theory of Spontaneous Generation - that living organisms could
develop from nonliving matter.
[This theory was challenged by the Italian physician Francesco Redi
(1626–1697)]
• Italian physician Francesco Redi placed meat in three
containers-Uncovered, covered with paper, covered with a
fine gauze to exclude flies .Then observed its ability to
produce maggots spontaneously.

Thus the generation of maggots by decaying meat resulted from

the presence of fly eggs, and meat did not spontaneously
generate maggots
Golden Era in microbiology

Louis Pasteur (1822-1895)

• Father of Medical Microbiology
• Developed swan neck shaped tubes to trap germs on
the walls of curved necks
• But when the necks were broken off microbial growth
commenced immediately
• Pasteur (1897) suggested that heating at 62.8 (145 F)
for 30 min. rather than boiling is enough to destroy
the undesireable organisms and also retaining the taste
of the product – Pasteurization
• Developed pasteurization, fermentation, effective
vaccines for anthrax and rabies
Robert Koch
• Demonstrated the role of bacteria in disease
• Perfected the technique of isolating bacteria in pure
culture
• He used gelatin to prepare the solid media. It failed
because-
• Gelatin is a protein , It gets digested by many bacteria
capable of producing proteolytic enzymes
• Also it melts when temperature gets above 25C
Fanne Eilshiminus Hesse (1850-1934)
• One of Koch’s Assistant proposed the use of Agar as the
solid media due to its higher melting points (96C) and
solidifying (40-45C) points
Koch’s Postulate
 Germ Theory
• The Germ theory of disease, also called the
pathogenic theory of medicine, is a theory
that proposes that microorganisms are the
cause of many diseases.
• Highly controversial when first proposed
• Germ theory was validated in the late 19th
century
• Now a fundamental part of modern medicine
and clinical microbiology
1843-1910
 Uses of Microbes
• Marine microorganisms form basis of food chain
• Use of soil microbes in recycling
• Role of microbes in photosynthesis
• Intestinal microbes help in digestion and synthesis of
vitamins
• Microbes in commercial applications- chemicals
(acetone),vitamins(B2, B12), enzymes, alcohols, and
many drugs.
• Role in food industry- vinegar, pickles, soy sauce,
cheese, yogurt, bread, and alcoholic beverages.
• Microbes are also manipulated to synthesize cellulose,
digestive aids, drain cleaner, therapeutic substances
such as insulin
Prokaryotes
 Prokaryotes : (Bacteria)
• Bacteria are unicellular
• Multiply by binary fission
• Differentiated on the basis of –Morphology
(shape), Chemical composition (staining
method), nutritional requirements, biochemical
activities, source of energy
• Shape : Coccus (spherical) , baccilus(rod shaped),
spiral (twisted)
• Diameter 0.2-2.0µm in diameter
• Length 2-8 µm
 Division in one plane

Division in
two plane

Division in
three plane

Division in
multiple
plane
 Prokaryotes : Structures external to cell wall
Glycocalyx- (capsule, slime layer , extracellular
polysaccharide)
• Gelatinious polysaccharide covering
• Capsules may protect pathogens from phagocytosis
• Capsules enable adherance to surfaces, help to provide
nutrition
Flagella :
• Long filamentous appendages consisting of a filament,
hook and basal bodies
• Help in locomotion
• Motile bacteria exhibit taxis-
+ve taxis----- towards attractant
-ve taxis is away from repellent
 Bacterial Flagella: various arrangements

Flagellar protein called H antigen distinguish between variations within species.

There are at least 50 different H antigens for E.coli
• Axial filaments
Spiral cells that move by the means of an axial filaments
(endoflagellam) are called Spirochetes
They are similar to flagellum except that they are wrap around
the cell

• Fimbriae and Pilli

Help cells to adhere to the surfaces
Pilli are involved in motility and DNA transfer
Pilli are longer than Fimbriae and are only 1-2 per cell
Axial Filaments Fimbriae
 Prokaryotes :Cell wall
• Surrounds the plasma membrane and protects cell from changes in
water pressure
• Bacterial cell wall consist of peptidoglycan, a polymer consisting of
NAG and NAM and short chains of amino acids
• Penecilin interfers with peptidoglycan synthesis
• Gram positive cell wall consists of many layers of peptidoglycan
• Gram negative bacteria have lipopolysaccharide- lipoprotein
phospholipid outer membrane surrounding a thin peptidoglycan
layer
• Outer membrane protects the cell from phagocytosis, Penicilin,
lysozyme and other chemicals
• Porins- are specific channel proteins that allow the allow the
exchange of small molecules across the outer membrane
Prokaryotes : Structure internal to cell wall
Prokaryotes : movement across membrane
 Diffusion

KMnO4 crystal in water

Osmosis
Cytoplasm

Nucleoid

Ribosomes
Endospores :
Eukaryotic cells
Eukaryotes : Cell wall and glycocalyx
 Eukaryotes : Plasma Membrane
• Eukaryotic plasma membrane consist of
phospholipid bilayer containing proteins- Intrinsic
and extrinsic proteins
• It also contain carbohydrates attached to proteins-
glycoprotein (which is lacking in prokaryotes)
• Eukaryotic cell move materials across plasma
membrane by passive process and active transport
(utilizing ATP), endocytosis (phagocytosis,
pinocytosis, receptor-mediated endocytosis)
• Cytoplasm – has a cytoskeleton and
chemically similar to prokaryotes

• Ribosomes- 80S ribosomes are found in

cytoplasm or attached to rough
endoplasmic reticulum
Eukaryotes : Organized Organelles
Difference between Gram+ve and Gram-ve bacteria
Archaea (Greek archaios means ancient)
• Best known for growing in restricted habitats eg.
Hypersaline or high temperatures
[Have special structural, chemical and metabolic
adaptations which enable them to grow in extreme
environments]
• Methanogenic and sulphate –reducing archaea have
unique cofactors that participate in Methanogenesis
• They are spherical, rod shaped, spiral, lobed,
triangular, plate shaped, irregular (Pleomorphic)
• Some are single cell, others form filamentous or
aggregates
• Range in diameter from 0.1-15µm (some filaments can
grow upto 200µm in length)
 Archaeal cell wall and membrane
• Can stain either gram +ve or gram –ve
• Some methanogenic archaea have pseudomeurine ( a peptidoglycan
like polymer thtat is cross-linked with L-amino acids)
• Have complex polysaccharide similar to Chondroitin Sulphate of
animal connective tissue
• Other distinctive feature of archae is membrane lipids
• They have branched chain hydrocarbons attached to glycerol by
ether-linkages
Thermophilic archaea sometimes link two glycerol groups to form long
tetraether chains
Diether chains- 20c long
Tetraether Chain- 40C long
• The chain lengths are adjusted by cyclizng chain to form
Penta cyclic rings
• Such penta cyclic rings are used by thermophilic archaea
in maintaining the liquid balance of membrane at high
temp. Also presence of phospholipid in memb
• Multiplication by binary fission, budding, fragmentation
or other mechanisms
• They can be aerobic, facultative anaerobic or strictly
anaerobic
• Nutritionally range from chemolithotrophs to
organotroph
• Include mesophiles, hyperthermophiles that can grow
above 1000C
 Ecology
• Archae are mostly found in “extreme
environments” like areas with high/low
temperature, pH, high conc, of salts, anoxic, dark,
under high pressure
[34% of prokaryotic biomass is due to Archaea in
Antartic coastal water]
• Some archaea are symbionts in digestive tract of
animals
• Archaeal gene sequences have been found in soil,
and tropical ocean surface water
 Genetics
• Some of archaeal genetics is similar to
bacteria and some resemble Eukarya
• Genome is smaller than that of bacteria
• Sign of archaeal diversity is due to the
variation in GC content (about 21%- 68%)
• Archaea have few plasmids
• Evidence of horizontal gene transfer between
thermophilic bacteria and archaea
• Comparative genomics – (completely sequence
genome of archaea, bacteria , eukarya revealed
that –
1. 30% of all genes are shared b/w archaea abd
eukarya which encoded for proteins involved in
transcription, translation and DNA metabolism
2. Large no. of genes shared by archaea and
bacteria and are involved in metabolic pathways
 Archaeal DNA replication
• Replication is bidirectional
• Replication origin is flanked by genes encoding
eukaryotic like initiation protein and few
archaea have multiple origin
• Some archaeal chromosomes are like bacteria
(like circular with single origin of replication)
whereas some have eukaryotic like histone
proteins that bind to form nucleolar like
structure
 Archaea: Metabolism
• Some are organotrophs, autotrophs,
phototrophs
• Little is known about biosynthetic pathways
Archaea is different in many ways from bacteria
and eukaryotes-
• Cell wall structure and chemistry
• Membrane lipid structure
• Molecular biology
• Metabolism
 Mycoplasmas
• Highly pleomorphic becoz lack cell wall and produce filaments that
resemble fungi (Mykes= fungi, Plasma=formed)
• Cells of genus mycoplasma are very small ranging in size from 0.1-
0.25 µm with cell volume that is only 5% of that of a typical bacillus
• Their size and plasticity allows them to pass through the filters that
retained bacteria. They were originally considered to be viruses
• They are smallest self-replicating organism that are capable of self
existance
• DNA studies revealed that genetically resemble gram positive
bacteria
• Eg M. Pneumoniae (cause common form of pneumoniae),
Spiroplasma (they have tight cork screw morphology , plant
pathogen, parasite of plant-feeding insects), ureaplasma (hydrolyze
urea in urine and cause urinary tract infections)
M.Pneumoniae
 Mycoplasma

• They can be grown in artificial media with

necessary nutrition and physical requirements
• The colonies are less than 1mm in diameter
and have characteristic “fried egg”
appearance when observed under microscope
• Fried egg colonies -Stain intensely with neutral
red or tetrazolium or methylene blue.
 Cyanobacteria (oxygenic
photosynthetic bacteria)
• They are named becoz of their blue-green (cyan)
pigmentation
• They do not carry oxygenic photosynthesis as
eukaryotic plants and algae do)
• They are capable of fixing nitrogen from atmosphere
• Nitrification :[Nirobacter oxidase convert ammonium
(NH4+) to nitrite (NO2-) which is further converted by
nitosomonas to nitrates (NO3-). This nitrate is
important in agriculture becoz this form of nitrogen is
easily mobile in soil and easily taken up by plants
• This activity is located in specialized cells called
heterocysts that can fix nitrogen
• Its species that grow in water contain gas
vaccoules which give them bouyancy
• Some move on solid surface by gliding mobility
• They are diverse and range from unicellular which
divide by binnary fission to colonial forms that
divide by mutiple fission to filamentous forms
that reproduce by fragmentation of filaments
• They are very important to environment
 Pleuropneumonia-like organisms
(PPLO)
• Now the name for Mycoplasma is pleuropneumonia
-like organisms (PPLO), broadly referring to organisms
similar in colonial morphology and filterability to the
causative agent (a mycoplasma)
• Several organisms with similar morphological
characteristics and cultural properties to
mycoplasmas have been isolated. These are
commonly referred to as pleuropneumonia-like
organisms or PPLO.
• Group of mycoplasmas produce extremely tiny
colonies on agar plates, and are called the T-strains.