MICROBIOLOGY AND PARASITOLOGY

Kathleen Jan M. Los Banez

BIO 300N
PRELIM REVIEWER  Eukaryotic
 Lacks cell walls
MICROORGANISM  Usually heterotrophic
 Unicellular
 Can reproduce asexually and sexually
MICROBIOLOGY  Moves by:
o Pseudopods
 specialized area of biology that deals with o Flagella
organisms too small to be seen without
o Cilia
magnification.
o Some are non-motile
PARASITOLOGY
FUNGI
 the study of parasites, their hosts, and the
 Eukaryotic
relationship between them.
 Cell wall has chitin
 Heterotrophic
 Unicellular (yeast) or
MICROORGANISMS
multicellular molds and
mushroom)
 Can reproduce asexually
 Linear DNA

HELMINTHS

 Flatworms and roundworms

 No cell wall
 Heterotrophic
 Can reproduce sexually and
BACTERIA asexually
 Have microscopic stages
 Prokaryotic
 Cell wall with peptidoglycan (most)
 Unicellular
 Reproduce by binary fission VIRUSES
(asexually)  Acellular
N. gonorrhoeae
 Circular DNA  Obligate intracellular
 Some are autotrophic, some are heterotrophic parasite
ARCHAEA  DNA or RNA
 May be enveloped or
 Prokaryotic naked
 Cell wall with lack peptidoglycan  A capsid (protein coat) is
 Unicellular required
 Reproduce by binary
fission (asexually)
 Extremophiles
o Thermophiles
o Halophiles
o Methanogens ALGAE

 Eukaryotic
 Cell wall has cellulose
 Photosynthetic
 Unicellular or multicellular
 Can reproduce sexually or
asexually
PROTOZOA  Often contains pigments:
green, red, or brown
 DARKFIELD MICROSCOPY

 Increases contrast without staining

by producing a bright image on a
MICROSCOPIC TECHNIQUES
darker background.
MAGNIFICATION VS RESOLUTION  Useful for viewing live specimens

PHASE CONTRAST MICROSCOPY

 Uses refraction and interference

caused by structures in the specimen
to create high contrast, high-
resolution images without staining.
 Useful for viewing live specimens,
and structures such as endospores
and organelles.

DIFFERENTIAL INTERFERENCE CONTRAST (DIC)

LIGHT MICROSCOPY MICROSCOPY

 MAGNIFICATION: up to about 1000×  Uses interference patterns to enhance contrast

 Uses visible or ultraviolet light to produce an between different features of specimen to produce
high-contrast images with 3D appearance.
image
 Useful for distinguishing structures within live and
TYPES unstained specimens and provide detailed
structures within cells.
 Brightfield microscopy
 Darkfield microscopy
 Phase contrast microscopy
 Differential interference contrast (DIC) microscopy
 Fluorescence microscopy
 Confocal microscopy

BRIGHTFIELD MICROSCOPY

 Commonly used in a wide variety of laboratory

applications as the standard method.
 Produces an image on a bright background

FLUORESCENCE MICROSCOPY

 Uses fluorescent stains to produce an image.

 Useful for identifying pathogens, to find particular
species, to distinguish living from
dead cells, or to find locations of
particular molecules within a cell

CONFOCAL MICROSCOPY

 Uses a laser to scan multiple z-planes successively,

producing numerous two-dimensional, high

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 resolution images at various depths that can be  Useful to observe the three-dimensional surface
constructed in a 3D image by a computer. details of specimens
 Useful for examining thick specimens such as
biofilms.

ELECTRON MICROSCOPY

 Magnification: 20,000-100,000×
 Uses electron beams focused with magnets to
produce an image.

TYPES

 Transmission Electron Microscopy (TEM)

 Scanning Electron Microscopy (SEM)

TRANSMISSION ELECTRON MICROSCOPY (TEM)

 Uses electron beams that pass

through a specimen to visualize small
images.
 Useful to observe small, thin
specimens such as tissue sections and
subcellular structures

SCANNING ELECTRON MICROSCOPY (SEM)

 Uses electron beams to visualize

surfaces.

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 EVOLUTION OF MICROBIOLOGY
≈3.5 BILLION YEARS AGO
 Microbially induced sedimentary structures were
found in Western Australia depicting earliest
record of bacterial community to exist.
 These microbial mats bind together sedimentary
grains to survive water current.
3180 B.C., 1122 B.C.
3180 B.C. - THE GREAT PESTILENCE
 Occurred during the reign of Pharaoh Mempses in
the First Dynasty of Egypt was the first recorded
epidemic in human history.
1122 B.C. - EARLIEST RECORD OF SMALL POX
 Caused by variola virus. Originated from China
and spread worldwide.
542 AD, 1347
THE BUBONIC PLAGUE
 Caused by bacteria Yersinia pestis transmitted by
rat flea.
 First epidemic occurred in the year 542 during the
Sasanian and Eastern Roman Empire killing 25-
50 million in the span of two centuries.
 Second epidemic killed 1/3 of European
population. Also called as "The Black Death"
LOUIS PASTEUR AND CHARLES CHAMBERLAND
MID-1600S
ZACHARIAS JANSSEN
 is credited with making one of the earliest
compound microscopes that has two lenses.
ROBERT HOOKE
 discovered the cell which heralded the Cell
Theory.
ANTON VON LEEUWENHOEK
 created a single-lens microscope which allowed
him to observe and provide accurate descriptions of
bacteria, protozoa, and fungi. He is also known
as the "Father of Microbiology"
1857-1867
LOUIS PASTEUR
 pioneered the formulation of Germ Theory of
Disease which states that microorganisms known
as pathogens or "germs" can lead to disease
(unsuccessful).
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 He also developed the process of pasteurization,
which became the basis of aseptic techniques.
 These theories and techniques were also used by
Joseph Lister to develop aseptic techniques during
surgery.
1876
ROBERT KOCH
 proved the Germ Theory of Disease which led to
formulation of Koch's Postulate.
 He successfully identified the following different
bacteria that caused anthrax, septicaemia,
tuberculosis, and cholera.
 He used revolutionary methods such as agar
culture and bacterial staining.
KOCH'S POSTULATES
 The microorganism must be found in abundance in
all organisms suffering from the disease but should
not be found in healthy
 organisms.
 The microorganism must be isolated from a
diseased organism and grown in pure culture.
 The cultured microorganism should cause disease
when introduced into a healthy organism.
 The microorganism must be re-isolated from the
inoculated, diseased experimental host and
identified as being identical to the original specific
causative agent.
1876
 used weakened dose of anthrax and cholera on
sheep and chickens to make them immune to the
disease.
 They went on to use this method to find a vaccine
for rabies (1885).
POST WORLD WAR II
1907 - PAUL EHRLICH
 formed an idea that it could be possible to kill
specific microbes (such as bacteria) without
harming the body itself.
 This lead to the formulation of Salvarsan, one of
the earliest antibiotic drug used for the treatment
of syphilis, caused by Treponema pallidum.
1928 - ALEXANDER FLEMING
 discovered penicillin, an antibiotic drug which
was responsible for enabling the control of many
 infectious diseases like tuberculosis, pneumonia,
and meningitis.

LATE 20TH CENTURY

1931

 The earliest electron microscope was developed.

1953

 DNA structure was discovered.

1977

 The first complete genomic sequencing of a

bacteriophage was accomplished.

2003

 The Human Genome Project was completed

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 TAXONOMY & CLASSIFICATION
TAXONOMY IS BASED ON THE ORGANISM'S

TAXONOMY GENOTYPE

● The orderly classification and grouping of ● genetic makeup of an organism

organisms into taxa (categories).
PHENOTYPE
● Involves three structured, interrelated categories:
nomenclature, classification, and identification. ● observable physical and functional features

EXAMPLES OF PHENOTYPIC CHARACTERISTICS

MACROSCOPIC MORPHOLOGY
● COLONY MORPHOLOGY
o the visual culture characteristics of a
bacterial colony on an agar plate.

MICROSCOPIC MORPHOLOGY
BACTERIAL NOMENCLATURE ● Refers to bacterial size, shape, arrangement (groups,
chains), and appendages.
● First word is the GENUS (capitalized)

● The second word is the SPECIES (not capitalized)

Both words are ITALICIZED.

STAINING CHARACTERISTICS
● Gram-negative (pink)

NAMES CAN ORIGINATE FROM ● Gram-positive (purple)

● the person who discovered it

● its appearance

● its natural environment or source

● the disease it causes

BIOCHEMICAL CHARACTERISTICS
● Nutritional requirements

● Antigenic markers

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 BACTERIAL MORPHOLOGY
● Susceptibility or resistance to antimicrobial agents
or chemicals
● Classification can be based on:
o ability to ferment
o carbohydrates
o carbon source used for growth
o enzymes present

 ANTIGENS
o a substance that triggers an immune response.
 SEROTYPING
o Classifying bacteria based on antigens present
using an antibody.
 Identifying the antigens present will allow us to
identify the bacteria.
 ANTIBIOGRAM PATTERNS
o classifying based on what type of antibiotics
bacteria are susceptible to.

BACTERIAL MORPHOLOGY

BACTERIAL SHAPES & ARRANGEMENTS

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 SPECIAL COMPONENTS OF GRAM-POSITIVE
CELL WALLS

TEICHOIC ACIDS

BACTERIAL CELL STRUCTURE  Function for the attachment of the organism to the
host cell.
 Provide tensile strength to the bacterial cell wall.

POLYSACCHARIDES

 Include neutral and acidic sugars.

 Play an important role as mediators of bacterial
interactions with the environment.

ENVELOPE STRUCTURES

BACTERIAL ENVELOPE
SPECIAL COMPONENTS OF GRAM NEGATIVE
● A complex multilayered structure that protect from CELL WALLS
their unpredictable and often hostile environment. OUTER MEMBRANE
● Composed of the outer membrane (glycocalyx), the
 Bi-layer structure composed of lipopolysaccharide
peptidoglycan cell wall, and the plasma membrane. (LPS).

GLYCOCALYX LIPOPROTEIN
 Outermost covering (external to the cell wall).
 Anchor the outer membrane to the peptidoglycan
 A GELATINOUS SUBSTANCE composed of layer and stabilizes the outer membrane.
polysaccharide or polypeptide, or both.
 CAPSULE PERIPLASMIC SPACE
o if strongly attached to the cell wall
 a fluid filled space between the outer membrane
 SLIME LAYER and the inner plasma membrane.
o if loosely attached  It contains enzymes for the breakdown of large
non-transportable molecules and enzymes that
detoxify and inactivate antibiotics

CELL WALL

 Principal component ➡️peptidoglycan

 Multi-layered in Gram (+) bacteria and single-
layered in Gram (-) bacteria.
 Protects the bacteria from osmotic damage and
plays an important role in cell division.

ACID-FAST BACTERIA CELL WALL

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 Acid-fast bacteria are gram-positive, but in addition  functions in cell division and is also involved in
to peptidoglycan, acid-fast cell wall contains large the secretion of substances produced by bacteria.
amounts of glycolipids, especially mycolic acids

(hydrophobic ➡️difficult to stain).

RIBOSOMES

 functions in protein synthesis.

 The hydrophobic nature of the cell wall protects the
bacteria from harsh chemicals such as strong acids GRANULES / INCLUSION BODIES
and detergents.
 serve for storage of food and energy.

PROJECTING STRUCTURES
FLAGELLA

 Thread-like structures that is made up of protein

flagellin. ENDOSPORES

 Function in bacterial motility.  Produced by some bacteria when they are placed in
a hostile environment.
PILI / FIMBRIAE  Composed of dipicolinic acid which confers
resistance to heat, drying, chemical agents, and
 Fine and short in comparison with flagella, and is
radiation.
made of protein called pilins.
 SPORULATION
 May also function in motility. o process of spore production.
 GERMINATION
COMMON PILI o spore begins to grow vegetative cells and
 for adherence to cell surface hyphae.

SEX PILI

 attachment to another bacterium during

conjugation.
AXIAL FILAMENTS

 Also called as end of flagella and are found

spirochaetes.

 Produce movement as it rotates which propel the

organism forward.

MICROBIAL GROWTH
REQUIREMENTS
CYTOPLASMIC MEMBRANE GROWTH
 Also called as the cell membrane or cell sac.
 Orderly and organized increase in the sum of all
 Selectively permeable.
components of the organism.
 In aerobic organisms, it is the site of the electron
 Entails the replication of all cellular structures,
transport chain and serves as the site of ATP
production. organelles, and components.
 Contains the enzymes needed for the biosynthesis  Bacterial growth

✅ Cell number
of DNA, cell wall components, and membrane
o
lipids
INTERNAL STRUCTURES o ❌ Organism size
NUCLEOID

 a chromatin-dense area that contains the bacterial

DNA, associated proteins, and RNA that are As in any living organism, microbes require certain nutrients
responsible for controlling the bacteria's activity and physical conditions that will promote their growth
and reproduction.
NUTRITIONAL REQUIREMENTS
MESOSOMES
CARBON

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 Carbon makes up the structural backbone or AEROBES
skeleton of all organic molecules.
 utilize molecular oxygen for energy production.
 Organisms can be classified based on carbon
source: OBLIGATE ANAEROBES
AUTOTROPHS (LITHOTROPHS)  microbes that cannot survive in the presence of
oxygen.
 utilize inorganic compounds (e.g., carbon dioxide)
and inorganic salts as their sole carbon source. FACULTATIVE ORGANISMS
HETEROTROPHS (ORGANOTROPHS)  can grow and survive under both aerobic and
anaerobic conditions.
 make use of organic substances like sugars or
glucose as their carbon source.
TEMPERATURE
NITROGEN, SULFUR, PHOSPHORUS  Enhanced enzyme activity requires certain
temperatures.
 Necessary for the synthesis of cellular materials
like proteins and nucleic acids.  Organisms can be classified based on their
temperature requirements:
 Nitrogen and sulfur are required for the synthesis
of proteins. THERMOPHILES
 Nitrogen and phosphorus are essential for the
synthesis of nucleic acids and ATP.  grow best at temperatures higher than 40 °C
 Approximately 14% of the dry weight of a
MESOPHILES
bacterial cell is nitrogen and about 4% is sulfur and
phosphorus.  require an optimal temperature of 20 °C–40 °C

PSYCHROPHILES
INORGANIC IONS
 require an optimum temperature of 10 °C–20 °C
MAGNESIUM

 stabilizes ribosomes, cell membranes, and nucleic pH

acids.
 Acidity or alkalinity of the bacterial environment
POTASSIUM affects growth.
 Organisms can be classified based on their pH
 required for the normal functioning and integrity of
requirements:
ribosomes.
ALKALOPHILES
CALCIUM
 grow best at pH 8.4– 9.0.
 an important component of gram positive bacterial
cell wall and contributes to the resistance of NEUTROPHILES
bacterial endospores against adverse environmental
conditions.  grow best in pH 6.5– 7.5

ACIDOPHILES

PHYSICAL REQUIREMENTS  grow best at pH < 6.0

MOISTURE/WATER
 The bacterial cell is composed mainly of water. It
serves as the medium from which bacteria acquire
their nutrients.
 It is essential for all living organisms. It functions
as a solvent, a temperature buffer and a metabolite
in living cells.
OSMOTIC CONDITIONS
 Determined by salt concentration which affects the
movement of solvent in and out of a permeable
membrane (e.g. cell membrane).
 Organisms can be classified based on their pH
requirements:

HALOPHILES
OXYGEN
 require high salt concentration for growth.
 Used by aerobic bacteria for cellular respiration.
OSMOPHILES
 Organisms can be classified based on their oxygen
requirements:

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 require high osmotic pressure for optimal growth.
(e.g. high sugar conc.)

BACTERIAL GROWTH CURVE

 Illustrates the phases in the growth of the

population of bacteria when they are grown in a
culture of fixed volume.

LAG PHASE

 Period of adjustment. No increase in cell number.

 Increased metabolic activity (1-4 hrs).

EXPONENTIAL PHASE

 Rapid cell Division.

 Doubling time is determined. (8hrs).

STATIONARY PHASE

 Period of equilibrium. Growth rate slows down.

Nutrients deplete.
 Waste accumulate.
 Bacterial death starts.
 Cell death = Living cells (months-yrs).

DEATH PHASE

 Period of rapid cell death.

 Cell death > Living cells. (hrs-days).

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