LESSON 1: Introduction to Microbiology Applied Microbiology

Microbiology- study of single-celled organism FIELD APPLICATION

and AGENTS too small to be seen clearly by the Medicine Control of infectious
naked eye. disease
Agriculture Microbial diversity
Microorganism- single celled organism and its effects on the
soil processes and
MICRON NANOMETER
productivity
Fundamental unit of Unit length in metric
Industry/Biotechnology Production of
length which is the system equal to one-
antibiotics, alcohols,
millionth part of billionth of meter.
enzymes and other
meter.
important chemical
AKA micrometer Represent by the
products
(𝛍𝐦) symbol of (nm)
1 micron =1 𝛍𝐦 = 1 nanometer= 1nm=
10^-6m 10^-9m Scope of Microbiology
1 micron = 1/1000 1nm = 1/1000 𝛍𝐦
mm 1.It is concerned with the form, structure,
physiology, reproduction, metabolism, and
identification of microorganisms.
Basic Specialization Under Microbiology
2.Includes the study of their distribution in
VIROLOGY- study of viruses nature.
BACTERIOLOGY- study of bacteria 3.Microorganisms as models in some biological
MYCOLOGY- study of fungi (e.g. mushrooms phenomenon.
and yeasts) 4.Utilization of microorganisms in the
PHYCOLOGY- study of algae (e.g. blue-green industries.
algae and cyanobacteria) Modern Era of Microbiology
PROTOZOOLOGY- study of protozoan Genomics

Transcriptomics
Scope of Microbiology Proteomics
Anatomy and morphology Metabolomics
Physiology and metabolism

Diversity Molecular Microbiology

Evolution 21th century – age of biology and
Ecology biotechnology

Application
Distribution of Microorganism in and on Earth -Developed acetone=butanol-ethanol
fermentation process, which produces
HABITAT PERCENT OF TOTAL
acetone through fermentation using
Marine subsurface 66
Clostridium acetobutylicum
Terrestrial subsurface 26
-1st president of Israel in 1949
Surface soil 4.8
Ocean 2.2
7. Medicine and related health science
All other habitat 1.1
Note: Estimated total number of bacterial cells: a. Pathogens (cause diseases)
2.5 x 1030 cells b. Antibiotics insulin, growth,
hormones, enzymes, vaccines
Importance Of Microorganism

1. Food production
FOOD/BEVERAGE MICROBE
Lactose-free milk Propionibacterium
freundenrechii
Wine Saccharomyces
cerevisiae
Saccharomyces spp.
Vinegar Acetobacter aceti
Nata de coco Acetobacter xylinum
Sour dough Saccharomyces
cerevisiae
Rice cake (puto) Streptococcus spp.
Angkak (red rice) Monascus purpureus
Tempeh (soybean, Rhizopus spp. Trivia
Indonesia) Erythromycin- discovered in 1949 by Dr,
Abelardo Aguilar from Streptomyces erythreus
2. Symbionts
Symbiodinium- coral association Eli Lilly Co. marketed the antibiotic as Ilosone in
3. Recycling of chemical elements in the 1952.
soil, water and air
8. Pollution control
4. Basis of food chain in the oceans, lakes
Bioremediation and biodegradation
and rivers
9. Genetic engineering/biotechnology Bt
5. Role in the photosynthesis
corn and other transgenic plants
Prochlorophytes- major contributor of
productivity in the open oceans
6. Commercial application
Poly-β-hydroxybutyrate (PHD)
And poly- β-hydroxyvalerate (PVD)
Copolymer for production of
microbial plastic

TRIVIA
Chaim Weizman- father of industrial
fermentation
 10. Microbial forensic demonstrating that maggots come from eggs of
11. Biofuels flies.
12. Research tools
a. Less space for growth, less
maintenance
b. Fast growth
c. Very wide range of biochemical
action
d. Their metabolic processes are found
in higher forms
e. Growth conditions can be altered
f. Can be observed in great detail
while undergoing metabolism
g. Versatile in their synthetic use John Needham (1745)- argued that microbes
arose spontaneously in broth from a “life force.”
LESSON 2: History and Developments in the (prove spontaneous theory)
Field of Microbiology Lazzaro Spallanzani (1729–1799)- disprove
1546 those of Needham. Life originates from a “life
force” that was destroyed during Spallanzani’s
Girolamo Fracastoro (1478- 1553) – disease was extended boiling. Any subsequent sealing of the
caused by invisible living creatures (germs of flasks then prevented new life force from
contagion; seminaria morbis- seeds of disease) entering and causing spontaneous generation.
1684 Rudolf Virchow (1858)-
Antoni Van Leeuwenhoek (1632-1723)- Theory of Biogenesis- living things can only
observed bacteria and protozoan using his come from other living things. (counter-
homemade microscopes hypothesis to spontaneous theory)
“The Golden Age of Microbiology” Louis Pasteur (1822-1895)-
Was driven by search for the ff. answers

1. Is spontaneous generation of microbial

life possible?
2. What causes fermentation?
3. What causes disease?
4. How can we prevent infection and
disease?

Spontaneous Generation Theory

- living organisms could arise from nonliving

matter (a) French scientist Louis Pasteur, who
definitively refuted the long-disputed theory of
Francesco Redi (1626-1697)- challenge the spontaneous generation. (b) The unique swan-
theory of spontaneous generation by neck flask feature used in Pasteur’s experiment
allowed air to enter the flask but prevented the pathogen must culture
entry of bacterial and fungal spores. (c) be grown in
Pasteur’s experiment consisted of two parts. In pure culture
the first part, the broth in the flask was boiled Cells from pure Experimental
to sterilize it. When this broth was cooled, it culture of the animal
remained free of contamination. In the second suspected
part of the experiment, the flask was boiled pathogen must
cause disease
and then the neck was broken off. The broth in
in a healthy
this flask became contaminated.
animal
Further Support against SGT The suspected Experimental
pathogen must animal
John Tyndall and Ferdinand Cohn- Tyndall's be reisolated
research emphasized the importance of heat- and shown to
resistant forms, and Cohn's discovery of be the same as
endospores provided insights into the the original
mechanisms by which some microorganisms
could survive adverse conditions.

Germ Theory of Disease

(Robert Koch)

It states that microorganisms known as

pathogens or "germs" can cause disease.

Joseph Lister (1827-1912)- found a way to

prevent infection in wounds during and after
surgery. He was the first to apply the science of
Germ Theory to surgery.

Robert Koch (1843-1910)- discovered the

bacteria that causes anthrax, septicaemia,
tuberculosis and cholera.

Koch’s Postulate Advances in microbiology by Koch

1. Simple staining techniques for bacterial cells

and flagella.
Postulate Tools
The suspected Microscopy, 2. 1st photomicrograph of bacteria.
pathogen must staining
3. 1st photomicrograph of bacteria in diseased
be present in
tissue.
all cases of the
disease and 4. Techniques for estimating the number of
absent from bacteria in a solution based on the number of
healthy colonies that form after inoculation onto a
animals.
solid surface.
The suspected Laboratory
5. The use of steam to sterilize growth media.
6. The use of Petri dishes to hold solid growth
media.
7. The use of agar as a solidifying agent.
8. Aseptic laboratory techniques such as
transferring bacteria b/w media with heat-
sterilized platinum wire.
9. Elucidation of bacteria as distinct species.
10.Development of nutrient broth and agar.
1798/1796
Key People in Microbiology
1798/1796
Edward Jenner- introduced a vaccination
procedure for smallpox
Variolation (Asia)- Dried smallpox scabs were
blown into the nose of an individual who then
contracted a mild form of the disease.
Ignaz Semmelweis- proposed that
handwashing could prevent the spread of
infection
Walther and Angelina/Fanny Hesse (suggested
replacing gelatin with agar) - early contributors
to bacteriology.
Gelatin- animal bones, cartilage, and skin
Gulaman-made of processed Gracilaria
seaweed; or carrageenan derived from other
farmed seaweed species like Eucheuma and
Kappaphycus alvarezii.
Richard Petri- inventing the device known as
the Petri dish.
Alexander Fleming (6 August 1881 – 11 March
1955)- discovered penicillin (a medication used
to manage and treat a wide range of infections)
Selman Waksman-he was awarded the Nobel
Prize in Physiology or Medicine for
"Ingenious, systematic, and successful studies
of the soil microbes” that led to the discovery of
streptomycin.
Martinus Willem Beijerinck- pioneer of general
microbiology, first discovered a pathogen that
was smaller than a bacterium. He called it a
virus and is therefore the father of virology.
Sergei Winogradsky- founder of modern
microbiology and the first microbial ecologist.
Molecular Microbiology
Carl Woese- uncovered the 'third domain of
life'. He achieved this by defining Archaea (a
group of single-cell prokaryotic organisms) – by
phylogenetic taxonomy of 16S ribosomal RNA,
a technique pioneered by him.
Thomas Brock- discovered temperature-
resistant bacteria named Thermus aquaticus
(best growth at 65–70 °C (149–158 °F), but can
survive at temperatures of 50–80 °C (122–176
°F)
Karl B. Mullis- invented the polymerse chain
reaction (PCR) method (a technique to make
many copies of a specific DNA region in vitro (in
a test tube rather than an organism)
LESSON 3: Microbial Evolution
Stromatolites- layered sedimentary formations
that are created mainly by photosynthetic
microorganisms such as cyanobacteria
Cyanobacteria- special categories of bacteria
that can perform photosynthesis.
 Endosymbiotic Hypothesis of Eukaryotic
Evolution

Origin of Cellular Life

1. Surface Origin Hypothesis-

2. Subsurface Origin Hypothesis- States that life

originated at hydrothermal springs on ocean
floor (hydrothermal vents)

Structure and Function of the RNA

passive genetic messengers responsible for:
1. transcribing genetic code from DNA
molecules
2. carrying that code to other sites in the cell
for the production of proteins.
LESSON 4: Introduction to Microbial
Systematics and Taxonomy
Systematic or phylogeny- study of evolution
history of origin
Taxonomy- put organism into categories or
taxa, to show degree of similarities

Domain-phylum-class-order-family-genus-
species

Parts/Branches of Taxonomy
1. Classification- arrangement of organism

Inti groups or taxa on the basis of mutual

similarity (phenetic classification) or
evolutionary relatedness (phylogenetic
classification)

NOTE: Phospholipid Membrane vesicle enclose 2. Nomenclature- assignment of names in

the replication machinery. taxa accordance to international rules
3. Identification- process of determining
Last Universal Common Ancestor that particular isolate belongs to a
(LUCA) Madigan recognized taxon

Taxonomic Approach
1. Phenotypic- based on observable
characteristics
 negative) to Staphylococcus (catalase
positive)
b. Test for indole- presence is
detected by adding Kovacs’ reagent.
Red color on the surface is positive an
orange-yellow color is negative test for
indole. Used to differentiate E-coli
(indole positive) Enterobacter (indole
negative)
c. Fermentation reaction
d. Bacteria sensitivity test
e. Analytical Profile Index (API) test strips

2. Phylogenetic- based on evolutionary

relationships rather than general
resemblance.
- use of evolutionary chronometers
such as rRNA sequences, etc

note: Gene encoding for the 16S ribosomal RNA

(rRNA) for prokaryotes, 18S rRNA for eukaryotes

RNA as the evolutionary chronometer

1. Universally distributed

2. Functionally constant

3. Sufficiently conserved

4. Adequate length to provide a deep view of

evolutionary relationships

3. Genetic or molecular – based on

organism’s genes, chromosomal gene
exchange, proteins, nucleic acids, etc

a. DNA-DNA hybridization

a. Slide catalase test – place a drop of

hydrogen peroxide. a positive shows
gas bubbles; a negative reveals an
absence of gas bubbles (used to
differentiate Streptococcus (catalase
b. DNA profiling Polyphasic taxonomy = phenotypic +
phylogenetic + genotypic
Taxonomic Rank
Note: Species relatedness: 97% or greater in 16S
Rank Example
rRNA and 70% or greater DNA-DNA
Domain Bacteria
hybridization
Phylum Proteobacteria
Class y-Protobacteria
Order Enterobacteriales
Family Enterobacteruaceae LESSON 5: The Major Divisions of Life Microbial
Genus Shigalla Diversity
Species S. dysenteriae

Sources of Carbon, Energy and Electron

Biological Species Concept
Carbon Autotrophs - CO2 Sole or
a group of interbreeding or potentially sources principal
interbreeding natural populations that are biosynthetic
reproductively isolated from other groups carbon source
Heterotrophs - Reduces,
Bacterial Species
performed organic
a collection of strains that share many stable molecules from
properties (phenotypic and genotypic) in other organism
common and differ significantly from other Energy Phototrophs - light
groups of strains. sources
Chemotrophs - oxidation of
Strain organic or
inorganic
a population of organisms that descends from compounds
a single organism or a pure culture Electron Lithotrophs - reduced inorganic
Biovar Source molecules
Organotrophs - organic molecules
variant strain characterized by biochemical or
physiological difference
Phototrophs- With pigments for conversion of
Morphovars light energy to chemical energy
variant strain characterized by morphological Two forms:
difference
1. oxygenated photosynthesis
Serovar (cyanobacteria and algae)
2. anoxygenic photosynthesis (purple
have distinctive antigenic properties
bacteria, green bacteria, heliobacteria)
Type Strain
Chemoorganotrophs- oxidize organic chemicals
strain of the species, usually one of those first for metabolism - obtain energy from organic
studied and often more carefully characterized compounds aerobically (with oxygen) or
anaerobically (without oxygen)
Chemolithotrophs- oxidize inorganic chemicals Fungi
for metabolism only among prokaryotes. Aspergillus
Alternaria
Domain Bacteria Mucor
Rhizopus
Trichoderma
Candida
Saccharomyces
Schizosaccharomyces
Kluyveromyces

Virus- submicroscopic infectious agent that

Phylum replicates only inside the living cells of an
Aquificae organism.
Thermotogae
Thermodesulfobacteria
Deinococcus- Thermus
Chrysiogenetes
Chloroflexi
Thermomicrobia
Nitrospirae
Deferribacteres
Cyanobacteria formerly known as
blue-green algae

Chlorobi
Proteobacteria Largest phylum of
bacteria known
Firmicutes
Planctomycetes
Chlamydiae
Spirochaetes
Tenericutes
Fibrobacteres
Acidobacteria
Bacteroidetes
Verrucomicrobia
Domain Archaea
 Subviral Particles Spirilium
Viroids- infectious RNA molecules (246 to 399
nucleotides) affecting plants (e.g. coconut
cadang-cadang viroid)
Spirillum volutans
Prions- infectious proteins affecting animals
Spirochete
(e.g. Creutzfeldt-Jakob Disease, Bovine
Spongiform, Encephalopathy)

LESSON 6: Cell Shape and Size Treponema pallidum

Budding and
Distinguishing Characteristics of The
appendaged bacteria
Prokaryotic Cells

1. DNA not enclosed within a membrane

2. Lack other membrane-bound organelles

Caulobacter crescentus
3. DNA not associated with histone proteins Filamentous bacteria
4. Cell walls almost always contain the complex

polysaccharide peptidoglycan

5. Reproduction usually by binary fission Chloroflexus aurantiacus

Shapes of Bacterial Cells Cossobacillus

Shape Bacteria
Coccus Staphy
lococc
us Haemophilus influenzae,
aureus Gardnerella vaginalis, and
Chlamydia trachomatis
Rod/ Bacillus Actinomycete
mycelium

Lactobacillus casei
Pleiomorphic
Vibrios

Vibrio Prahaemolyticus
Mycoplasma pneumoniae
 Hyphomicrobium Sarcinae
with hyphae and bud

Sarcina ventriculi, Sarcina

ureae
Staphylococci
Square bacterium

Staphylococcus aureus
Single bacillus
Haloquadratum walsbyi
Star-shaped bacteria

Stella humosa, Stella Coxiella burnetii, Moraxella

vacuolata bovis, Klebsiella
rhinoscleromatis
Diplobacilli
Cell Arrangement

Cell Arrangement
Diplococci
Streptobacilli

Moraxella catarrhalis,
Neisseria gonorrhoeae
Streptococci Streptobacillus moniliformis
Palisades

Streptococcus pneumoniae
Tetrad

Aerococcus, Pediococcus and Corynebacterium

Tetragenococcus diphtheriae
 Prokaryotic cells are small LESSON 7: Functional Anatomy of the
Prokaryotic Cell (Cell Wall)
1.Higher surface-to-volume ratio
Cell Wall
2.Higher growth rate
complex, semirigid structure composed of
3.Higher mutation rate
macromolecular network called the
Cell size and volume of representative peptidoglycan (murein)
prokaryotic cells
Responsible for:

1. Characteristic shape of the cell

2. Protection from osmotic lysis

3. Point of anchorage for flagella

4. Contribute to pathogenicity

5. Protection from toxic substances

6. Site of action of several antibiotics

7. Used to differentiate major types of bacteria

Epulopiscium Gram positive Gram negative
fishelsonii (Purple) (Pink)
Staphylococcus aureus Escherichia coli

Thiomargarita
namibiensis

Peptidoglycan (murein)- polysaccharide

composed of two sugar derivatives (N-
acetylglucosamine and N-acetylmuramic acid)
Mycloplasma
and amino acids (L-alanine, D-alanine, D-
genitalium
glutamic acid, Lysine/Diaminopimelic acid
(DAP)

Gram-positive CW

consists of 20 to 80 nm thick homogenous

Nanoarchaeum peptidoglycan or murein layer outside the
equitans plasma membrane.
(400 nm)
Also contain large amounts of teichoic acids
(lipoteichoic acids/wall teichoic acids)-polymers
of glycerol or ribitol joined by phosphate groups
 Gram-negative CW LESSON 8: Functional Anatomy of the
Prokaryotic Cell
consists of 1 to 3 nm peptidoglycan layer
surrounded by a 7 to 8 nm thick outer Structures internal to the CW
membrane (lipopolysaccharide; LPS)
1. Plasma (Cytosplasmic) membrane –
thin structure lying inside the CW and
enclosing the cytoplasm of the cell

Functions:

1. selectively permeable barrier

2. mechanical boundary of cell

3. nutrient and waste transport

4. location of many metabolic processes

5. detection of environmental cues for

chemotaxis
Periplasm – loose network of peptidoglycan;
gel-like consistency

- contains hydrolytic enzymes, binding proteins

for nutrient processing and uptake,
chemoreceptors.

Porins - transmembrane proteins as channels

for solutes
2. Cytoplasm (cytoplasmic matrix) –
substance lying between PM and
nucleoid- 70- 80% water containing
proteins, carbohydrates, lipids, Aquaspirillum magnetotacticum
inorganic acids and low molecular
weight compounds 4. Nuclear area (nucleoid; nucelar body;
3. Inclusion bodies (Cell inclusions) – nuclear region; chromatin body)-
granules of organic or inorganic irregularly shaped region composed of
materials; storage of carbon, phosphate 60% double-stranded DNA, 30% RNA
and other substances: and 10% protein
A. metachromatic granules (coll. volutin)-
reserve of inorganic phosphates
B. polysaccharide granules- glycogen and
starch
C. lipid inclusions- –ex. poly-β- 5. Internal membrane structures –
hydroxybutyric acid membranous structures
D. sulfur granules
E. carboxysomes- contain the enzyme mesosomes – invaginations of the PM in the
ribulose 1,5-diphosphatecarboxylase shape of vesicles, tubules or lamellae
F. gas vacoules- aggregates of enormous
numbers of small, hollow, cylindrical
structures called gas vesicles
G. cyanophycin granules- stores exta
nitrogen
H. magnetosomes – contain magnetites
(Fe3O4), greigite (Fe3S4) pyrite (FeS2)

6. Ribosomes – very complex structures

made up of protein and rRNA; site of
protein synthesis

NOTE: prok: 70S R – 30S / 50S

 7. Endospores – special resistant, Types
dormant structures
1. Capsules – well organized and not easily
washed off

- protect bacteria against desiccation,

phagocytosis; adherence to surface

- may be a source of nutrition

Spores
Terminal spores

2. Slime layers – zone of diffuse, unorganized

materials that is easily removed

Subterminal pores 3. Flagella – filamentous appendages that

propel bacteria

Types
monotrichous

Central spores

Lophotrichous (tuft)
LESSON 9: Functional Anatomy of the
Prokaryotic Cell

Structures external to the CW

1. Glycocalyx – general term used for

substances that surround the cell

- viscous, gelatinous polymer composed of

polysaccharides, polypeptides or both

- primarily for attachment to surfaces

peritrichous 4.Fimbriae – thinner than flagella;
consists of proteins called pilin; at the
poles of cells or evenly distributed over
the entire surface; attachment to surfaces

Amphitrichous-

5. Pili – longer than fimbriae; about 1 to

Movement 10 per cell; join bacterial cells for the
a. Peritrichous transfer of DNA from one cell to
another

b. Polar

Gliding motility
Slime extrusion

3. Axial filaments- bundles of fibrils that

arise at the ends of the cell beneath the outer Oscillatoria (cyanobacteria)
motility proteins
sheath and spiral around the cell

- allows motility for corkscrew movements of

spirochetes
Flavobacterium johnsoniae
type IV pili;
protein adhesion
complex

Myxococcus xanthus
 Microbial Taxes
1. Chemotaxis- chemical stimulus
2. Phototaxis- light
3. Aerotaxis- air
4. Osmotaxis- oxygen
5. Hydrotaxis- water
6. Magnetotaxis- magnet

LESON 10: The Fungus as an Organism

Homokaryotic – nucleus is generally identical

(derived from a single unnucleated spore)

Heterokaryotic- usually a result of anastomosis

(fusion) and therefore the nucleus are
genetically different

Dikaryotic- with two distinct nuclei (in contrast

with monokaryotic)

Dimorphic- fungi which can switch between

mycelial or yeast forms

Pseudohyphae- intermediate forms between

yeast and hyphae form

Thallus- body of the fungus

Hyphae- branching filaments of fungal body

Note: mainly chitosan
Mycelia- collection of hyphae

Yeast- unicellular
 Strand type
(Sclerotinia spp)

ergots of rye
(Claviceps purpurea)

Hyphal Aggregates

Hyphal Aggregates

Mycelial strands

Rhizomorphs

Sclerotium

Common ancestor
Chytridiomycota
(were probably the
Types of Sclerotia first to branch away)

Types
Loose type
(Rhizoctonia species)
Allomyces, water
molds,

Terminal type
(Botrytis cinerea)

(Rhizopus, bread
molds, Mucor
Basidiomycota Sexual Fruiting Bodies
are most closely (sporocarp) of
related to each other ascomycetes (cup
than to the other fungi)
phyla [=-60 Cleistothecium

Mushroom, rusts,
smuts
Ascomycota

Neurospora, yeast, sac Perithecium

fungi

Apothecium

Pseudothecium

Cleistothecium- is a fully-enclosed fruiting

structure. These
typically have bag-like asci; some split open to Ascobolus
release their spores and are called furfuraceus
chasmothecia

Chlorioactis geaster

Perithecium- flask-shaped fruiting structures, Pseudothecium- asci are produced in a cavity or

often microscopic and/or embedded within locule buried within astroma of fungal mycelium
either the substrate it is fruiting in or a fungal
structure called a stroma.

Podospora

Ventiria inaequalis

Xylaria

Apothecium- cup-shaped with the hymenium

fully exposed (lining the interior of the cup),
though this can be inverted and take on strange
shapes Aspergillus- genus consisting of several hundred
mold species
Microstoma
floccosum

Pithya vulgaris
Phialides- sporogenous cells that produce Coleosporium
conidia through a specialized apical budding
process

Symmetrospora
oryzicola

Ustilago maydis

Exobasidium

Moniliella sp.

Sexual fruiting bodies of basidiomycetes

(basidiocarps)
Wallemia

Phallus

Examples of
Basidiomycota
Puccinia iridis Clavaria

Phleogena
Amylostereum
faginea
Clavariadelphis Asexual Spores

Spores
Chlamydospores- large
and
Pycnoporus thick-walled

Blastospores–
Russula
produced by budding

Boletus
Microconidia-
uninucleate,
non-septate, hyaline,
lunate with thin
Lycoperdon cell wall

Macroconidia- septate
Phialospores - spores
that form on
the tip of the phialide
of a fungus
Schizophyllum commune-

Porospore- the
developing spore
emerges through a
distinct ‘pore’ in
the hyphal wall
 LESOON 11: Culture Media and Pure Culture
Techniques

Culture- strain or kind of organism growing in

laboratory medium

Culture media- solution of various nutrients

suitable for the growth of a microorganism

Pure culture- culture containing a single kind of

microorganism

Culture media classifications based on:

Pilobolus kleinii a. Physical state

1. liquid- water-based solution without

solidifying agents

- broth, milk, infusion

2. semisolid- – w/ clotlike consistency at

ordinary room temp; w/ small amount of
solidifying agent (agar or gelatin) but does not
form firm substance

3. solid- w/ firm surface (solidifying agents:

agar, gelatin, silica gel, etc)
Glomeromycota- newly established phylum that
comprises about 230 species, all of which are
involved in close associations with the roots of
trees.

A good solidifying agent:

• not utilized by microorganisms

• does not inhibit microbial growth
• does not liquefy at room temperature

b. Chemical composition

1. Synthetic or defined media– compositions

are chemically-defined; composed of organic
and inorganic compounds with molecular
contents specified by means of exact formula
2. Non-synthetic or complex media- with produce yellow colonies with yellow zones
ingredients that are not chemically-defined, not
-Coagulase-negative staph produce small pink
simple or pure compounds and with no exact
or red colonies w/o color change to the medium
chemical formula
Blood Agar -5% sheep blood lysed by
hemolysins (exotoxins) –

hemolysis patterns:

- Beta– complete

Alpha hemolysis– partial

Gamma- none

Eosin Methylene Blue Agar- selective and

differential medium used to isolate fecal
coliforms (metabolize lactose in the media)

Chocolate agar- for growing fastidious

respiratory bacteria which need growth factors
such as nicotinamide adenine dinucleotide
(factor V or NAD) and hemin (factor X), which
are inside red blood cells

c. Functional type

Mac Conkey Agar- to isolate and differentiate

enterics based on their ability to ferment
lactose

Mannitol Salt Agar- Coagulase-positive staph

Pure culture – population of cells arising from a
single cell
Contaminants – unwanted microorganisms
Plating techniques
1. Streak plate
2. Spread plate
3. Pour plate
LESSON 12: Requirements for Microbial
Growth
Requirements for Microbial Growth
1. Chemical - carbon and nitrogen, minerals,
organic growth factors
2. Physical – temperature, pH, solutes and
water activity. Oxygen concentration, pressure,
radiation
3. microbial interaction
Nutrients – substances used in biosynthesis and
energy production
Macroelements/macronutrients – required by
organisms in large amounts
Growth factors – organic compounds required
in only very small amounts
Physical Requirements for Growth
1. Temperature
2. pH
Most bacteria and protozoa – neutrophiles
Fungi and algae – prefer slightly acidic
environment
3. Solutes and water activity
Xerophiles – can grow under very dry
conditions
Osmophiles – will grow at high sugar
concentrations/ tolerant of high NaCl
concentrations
Osmotolerant organisms – grow best at high
water activity but also tolerant of high sugar
concentrations
4. Oxygen – (Catalase test?)
Obligate aerobes require oxygen to live;
they are growing at the top of the tube of
nutrient broth.
Facultative anaerobes are bacteria that
grow with or without oxygen but grow best
in oxygen, with most growth near the top of
the tube. These bacteria are growing
throughout the tube of nutrient broth.
Obligate anaerobes do not grow in the
presence of oxygen, shown growing at the
bottom of the tube.
Aerotolerant anaerobes bacteria are not
affected by oxygen levels and can grow
equally well in aerobic or anaerobic
conditions (and anything in between).
Microaerophilic bacteria grow best in low
concentrations of oxygen, just below the
surface of the nutrient broth in the tube.
 Facultative bacteria – neither aerobic nor
anaerobic
Methods of culturing microorganisms – The
a. facultative aerobe – prefers anaerobic Five “I” s
conditions but grows aerobically
1. Inoculation
b. facultative anaerobe – prefers oxygen
rich conditions but grows anaerobically 2. Incubation
c. Obligate aerobes and facultative
anaerobes – w/superoxide dismutase 3. Isolation
and catalase 4. Inspection
d. Aerotolerant microorganisms – may
lack catalase, almost always have 5. Identification
superoxide dismutase
e. All strict anaerobes – lack both
enzymes or have them in very low
concentrations
5. Pressure

barotolerant – survive and adapt to extreme

pressure

barophilic – grow rapidly at high pressures

6. Radiation-radiation resistant life form

(Deinococcus radiodurans)

Constraints in working with microorganisms:

1. Habitats with complex associations of

microbes

2. Must be grown in artificial conditions

3. Invisible and widely distributed