MICR 21 LEC

MICROBIOLOGY AND
PARASITOLOGY MAJOR CATEGORIES OF MICROBES
MIDTERM REVIEWER
Acellular Microbes
- also called infectious particle (virus,
MICROBIOLOGY AND prions)
PARASITOLOGY: INTRODUCTION
Cellular Microbes
- include all bacteria, all archaea, some
MICROBIOLOGY algae, all protozoa, and some fungi.
- the study of the dominant form of life on
Earth — Microbes. Samples
Pics
Microorganisms
- Life forms too small to be seen by the
unaided human eye.
- Many are undfifferentiated single-celled CATEGORIES OF MICROBES
organisms, but some can form complex
structures, and some are even multicellular. Organisms and biological entities studied by
- Typically live in complex microbial microbiologists can be
communities
Cellular includes:
STUDYING MICROBIOLOGY Fungi - eg. yeasts, molds
- microscopy is foundational to microbiology Protists - e.g. algae, protozoa, slime molds
Microscopy is the use of a microscope, thus allows us to Bacteria - e.g. Escherichia coli
investigate certain microscopic forms of life. Archaea - e.g. Methanogens
- the cultivation of microorganisms is also
foundational to microbiology Acellular includes:
Viruses - composed of protein and nucleic acid
Microbial Culture Viroids - composed of RNA
- collection of cells that have been grown in Satellites - composed of nucleic acid enclosed
or on a nutrient medium in a protein shell
Prions - composed of protein
Medium
- a liquid or solid nutrient mixture that
PATHOGENS
contains all of the nutrients required for a
- disease-causing microorganisms
microorganism to grow - only about 3% of known microbes are
pathogenic
Colony
- visible group of microorganism grown from Vast majority of known microbes are
a media nonpathogens — microbes that do not cause
disease.
MICROBIAL CELLS
- PICTURE

Graciel Aronce
BSM 2-3
 HISTORY OF MICROBIOLOGY

Cyanobacteria (3.5 bya - billion years ago)

- First microorganism on earth

EARLIEST KNOWN INFECTIOUS

DISEASES
• The earliest known account of a
“pestilence” occured in Egypt about 3180
translation: “ang dumi mong bata ka”
BC
• Around 1900 BC, near the end of the
IMPORTANCE OF STUDYING Trojan War, the Greek army was decimated
MICROBIOLOGY by an epidemic of wat is thought to have
been Bubonic plague
• INDIGENOUS MICROBIOTA • A disease thought to be smallpox occurred
- nagkakaroon ng kaalaman sa mga in China around 1122 BC.
microorganisms na nasa katawan ng tao • Epidemics of plague occurred in Rome in
- origin 790, 710, and 640 BC and in Greece
around 430 BC.
• ENVIRONMENTAL IMPACT
- Syphilis - Sexually Transmitted Infection
caused by Spirochete Treponema
• BIOTECHNOLOGY Pallidum
• It made its first appearance in Europe in
• INFECTIOUS DISEASES AND 1493
MICROBIAL INTOXICATIONS • Many people believe that syphilis was
carried to Europe by Native Americans who
were brought to Portugal by Christopher
NOTE: Columbus
Invitro - laboratory growth • The name “syphilis” was not given to the
Invivo - animals disease until 1530

THE EBERS PAPYRUS

INFECTIOUS DISEASE MICROBIAL
INTOXICATION • Describing epidemic fevers, was
discovered in a tomb in Thebes, Egypt
A pathogen colonizes a A pathogen produces a • It wa written around 1500 BC
person’s body. toxin in vitro.
| |
The pathogen causes a A person ingests the
disease toxin causes a disease
| |
PIONEERS IN THE SCIENCE OF
This type of disease is This type of disease is MICROBIOLOGY
known as an infectious known as a microbial
disease. intoxication
Anton Van Leeuwenhoek (1632-1723)

Graciel Aronce
BSM 2-3
- Father of Microbiology, Father of - Discovered the infectious agents that
Bacteriology, Father of Protozoology caused silkworm diseases
- As a hobby, he ground tiny glass lenses, - Made significant contributions to the germ
which he mounted in small metal frames, theory of disease—the theory that specific
thus creating what today are known as microbes cause specific infectious diseases
Single-lens microscope or simple (Bacillus anthracis & Mycobacterium
microscopes tuberculosis)
- The letters of his recorded observation he - Championed changes in hospital practices
sent to the Royal Society of London finally to minimize the spread of disease by
convinced scientists of the late 17th century pathogens.
of the existence of microbes. - Developed vaccines to prevent chicken
cholera, anthrax, swine erysipelas (a skin
”My method for seeing the very small animalcules I do disease), rabies in dogs and successfully
not impart to others; nor how to see very many
used to treat human rabies (Joseph
animalcules at one time. This I keep for myself alone.”
Meister).
The THEORY OF SPONTANEOUS
SWAN-NECK FLASK EXPERIMENT
GENERATION (Abiogenesis)
- His experiment that debunked the Theory
- On the basis of observation, many of the
of Spontaneous Generation
scientists of that time believed that life could
develop spontaneously from non-living
- insert diagram -
material (decaying corpses, soil, and
swarm gases)
“life only comes from life”
- For more than 2 centuries, from 1850, this
theory was deebated and tested.

THE THEORY OF BIOGENESIS

(CONTINUOUS GENERATION)
Robert Koch (1843-1910)
- First proposed by a German Scientist
German physician
named Rudolf Virchow in 1858.
- Made numerous contributions to the
- Life can only arise from pre-existing life
science of microbiology.
- Proved that the anthrax bacillus (B.
anthracis), which had been discovered
Louis Pasteur (1822-1895)
earlier by other scientists, was truly the
French Chemist
cause of anthrax
- His contribution are considered by many
- Discovered that B. anthracis produces
people to the foundation of the science of
spores, capable of resisting adverse
microbiology and a cornerstone of modern
conditions.
medicine
- Develop methods of fixing, staining, and
- Alcoholic fermentation
photographing bacteria
- Forms of life that could exist in the
- Developed methods of cultivating bacteria
absence of oxygen (“aerobes” and
on solid media
“anaerobes”)
> R.J. Petri - invented petri dish
- Pasteurization

Graciel Aronce
BSM 2-3
> Frau Hesse - suggested the use of agar
pathogen
(a polysaccharide obtained from seaweed) must be
as a solidifying agent. reisolated
- He and his colleagues established an and shown
experimental procedure to prove that a to be the
specific microbe is the cause of a specific same as the
infectious disease. This scientific procedure, original.
published in 1884, be came known as
Koch’s Postulates.

KOCH’S POSTULATES

Theoretical aspects Experiment PARASITOLOGY

al aspects - scientific discipline concerned with the
Postulates: Laboratory study of the biology of parasites and
Tools parasitic diseases.
1. The
suspected Medical Parasitology
pathogen Microscopy, - concerned primarily with parasites of
must be staining
humans and their medical significance, as
present in
all cases of well as their importance in human
the disease communities
and absent Laboratory
from healthy cultures PARASITES: Mode of Development

2. The Endoparasite (Ascaris Lumbricoides)

suspected
pathogen Experiment
must be al animals Ectoparasite (Dog tick)
grown in
pure culture. Erratic Parasite (Entamoeba Histolytica)

3. Cells Obligate Parasites (Head louse)

from a pure Laboratory
culture of reisolation
Facultative Parasite (Naegleria Fowler)
the and culture
suspected
pathogen Accidental Parasite (Dipylidium Caninum)
must cause
disease in a Permanent Parasites (Sarcoptes Scabies)
healthy
animal.
Temporary Parasite (Leeches)
4. The
suspected Spurious Parasite (Monocystis Lumbrici)

Graciel Aronce
BSM 2-3
 - considered as the most interesting
observations made in the Middle Ages
IMPORTANCE OF STUDYING - He gave the first recognizable description
PARASITOLOGY of the liver fluke

• PUBLIC HEALTH
• AGRICULTURAL AND ECONOMIC
IMPACT PIONEERS IN THE SCIENCE OF
PARASITOLOGY

RECORDED IDENTIFICATION OF Francesco Redi (1626-1697)

PARASITES Italian Physician (FATHER OF
PARASITOLOGY)
Egyptian Ebers Papyrus (c. 1550 BC) - One of the most important researchers of
- Describes the larger ectoparasites and the 17th century
what is now recognized as Dracunculus - He described and drew lice and ticks from
medinensis (guinea worm) nematodes in animals and human, and described the
humans, but without understanding their life donkey louse
cycles. - Refuted the concept of spontaneous
generation with a very simple but highly
Corpus Hippocratorum (5th and 4th effective experiment
Century BCE)
Hippocrates of Cos (c. 460-370 BCE) ”Everything which we know in past or present
times…came solely from the true seeds of the plants
- provides the first scientific observations
and animals themselves.”
about the clinical perception and treatment
of helminthic diseases. Edward Tyson (1651-1708)
English Physician
- performed a detailed examination of
De Lumbricis (6th Century AD) Ascaris lumbricoides which showed that
Alexander of Tralles (c.526-605) there were two sexes
- Deals with the topic of intestinal worms - His conclusion that sexual reproduction
- Fundamental work in the history of early took place in such simple animals was an
parasitology observation that again rejected the idea of
spontaneous generation in helminths.
De Animalibus (1478)
Albert Magnus (c. 1200-1280) Ronald Ross (1857-1932)
- recorded parasitic worms in horses, British army surgeon
dogs, falcons, and fish - elucidated the whole of the transmission
cycle in culicine mosquitoes and birds
A Newe Treate or Treatise Most infected with Plasmodium relictum in 1897.
Profytable for All Husbandmen (1532)
Charles Louis Alphonse Lavéran
Sir Anthony Fitzherbert (c. 1470-1538) (1845-1922)

Graciel Aronce
BSM 2-3
French physician, pathologist, and
parasitologist Nucleocapsid - Consists of a genome of
- discovers the causative parasites in the either DNA or RNA, surrounded by a capsid
blood of malaria patients in 1880. (protein coat), which is composed of many
- Traité des fiévres palustres small protein units called capsomeres.
- won the Nobel Prize for Physiology or
Medicine in 1907.
FOUR DIFFERENT TYPES OF VIRUSES
BASED ON STRUCTURE

MICROBIAL DIVERSITY: ACELLULAR Helical

AND PROKARYOTIC MICROBES - shaped much like hollow tubes with protein
walls.
- A single type of protomer associates
● Acellular Infectious Agents
together in a helical or spiral arrangement to
● The Domain Bacteria
produce a long, rigid tube
● The Domain Archaea
- The RNA genetic material is wound in a
spiral and positioned toward the inside of
Acellular Microbes
the capsid where it lies within a groove
- Lacks cellular structure and cellular
formed by the protein subunits.
organization
- Devoid of life
Icosahedral
- Pathogenic agent
- more prevalent than the helical
architecture
ex. Virus, viroids, prions
- made of 5 or 6 protomers
- constructed from ring- or knob-shaped
Virus
units called capsomers
Virus - venom or poisonous fluid
- resembles the geometric figure
- Infectious acellular entities that are
icosahedron
metabolically inert
- Obligate intracellular parasites
- Lack genes and enzymes necessary for
Spherical
energy production
- roughly spherical but somewhat variable
- Infectious microbe consisting of a segment
shape even though their nucleocapsid can
of nucleic acid (either DNA or RNA)
be either icosahedral or helical.
surrounded by a protein coat (capsid).
- enclosed in a membrane known as
- Extremely small and simple in structure
envelope (Lipid membrane)
(10 to 300 nm or 1 micrometer e.g. Ebola
- typically less virulent
virus)

Complex
Virions
- have capsid symmetry that is neither
- The simplest complete virus particles
purely icosahedral nor helical
- Specifically refer to the virus particle
- possess tails and other structures
existing outside the host cell
- Dorman state (Latency)

Graciel Aronce
BSM 2-3
- have complex, multilayer walls
surrounding the nucleic acid Categories of Bacteriophages:
Post-Bacterial Cell Invasion

ORIGIN OF VIRUS Virulent Bacteriophages

Theory No. 1 - Lytic cycle
- Virus existed before cells - The totality of the process takes less than
1 hour (<1 hr)
Theory No. 2
- Viruses represent ancient derivatives of Temperate Phages (Lysogenic Phages)
degenerate cells or cell fragments - Do not immediately initiate the lytic cycle,
but rather, their DNA remains integrated into
Viruses are NONLIVING ENTITIES the bacterial cell chromosome, generation
after generation.
Bacteriophages
Phagein - eat LYTIC CYCLE
- specifically infect and replicate within (APBAR)
bacteria cells Attachment, Penetration, Biosynthesis,
- obligate intracellular parasites Assembly, Release
- the most abundant organisms in the
biosphere Attachment
- utilized to treat illnesses that are caused O minutes
by a variety of bacteria - Absorption of phage to the surface of the
- Transduction bacterial cell.
- Attached to a protein or polysaccharide
Three Categories of Bacteriophages: molecule (receptor) - Strain specific
Based on Morphology |
Penetration
Icosahedron Bacteriophages 1 minute
- An almost spherical shape with 20 - Phage injects is DNA into the bacterial
triangular facets cell, acting much like a hypodermic needle
- The smallest icosahedron phages are - The capsid remains on the outer surface of
about 25 nm in diameter. the cell
- Taking over of the host’s cell’s “Machinery”
Filamentous Bacteriophages |
- Long tubes formed by capsid (protein coat) Biosynthesis
proteins assembled into a helical structure 10 minutes
- can be up to about 900 nm long - Phage genes are expressed, resulting in
the production of viral pieces.
Complex Bacteriophages - The host’s cell’s enzymes, nucleotides,
- Icosahedral heads attached to helical tails amino acids, and ribosomes are used to
- may also possess base plates and tail make viral DNA and viral proteins.
fibers |
Assembly

Graciel Aronce
BSM 2-3
 25 minutes - Viral nucleic acid “dictates” what occurs
- Viral pieces are assembled to produce within the host cell
virions.
- Viral DNA is packaged up into capsids Biosynthesis
| - Many viral pieces (viral nucleic acid and
Release viral proteins) are produced.
30 minutes - Some do not initiate biosynthesis right
- The host cell bursts open and all of the away, but rather, remain talent within the
new virions escape from the cell (lysis) host cell for variable periods.
- 50-1,000 new virions
Assembly
- Fitting the virus pieces together to produce
complete virions.

Release
- Some escape by destroying the host
leading to cell destruction and some of the
symptoms associated with infection with
that particular virus.
- Budding (Enveloped virus by pieces of cell
membrane)
Animal Virus
- Viruses that infect humans and animals
- May consist solely of nucleic acid
surrounded by a protein coat (capsid) or Inclusion Bodies
may be more complex - Remnants or collections of viruses
- Often seen in infected cells and are used
Multiplication of Animal Virus as a diagnostic tool to identify certain viral
(APUBAR) diseases.
Attachment, Penetration, Uncoating, - Found in the Cytoplasm (cytoplasmic
Biosynthesis, Assembly, Release inclusion bodies) or within the nucleus
(intranuclear inclusion bodies), depending
Attachment on the particular disease
- Attached to cells bearing appropriate
protein or polysaccharide receptors on their Latent Virus Infection
surface. - The virus remain latent for extended
period and persist within the host cell
Penetration - Limited by the defense system of the
- Enters the host cell, sometimes because Human body (Phagocytes and Interferon
the cell phagocytizes the vitus produced by virus-infected cells)
- e.g. Varicella-zoster virus - Shingles
Uncoating
- Viral nucleic acid escapes from the Antiviral Agents
capsids (protein coat)

Graciel Aronce
BSM 2-3
- Developed to interfere with virus-specific - Usually transmitted via insects, mites,
enzymes and virus production by either nematodes, infected seeds, cuttings, tubers,
disrupting critical phases in viral cycles or and contaminated tools.
inhibiting the synthesis of viral DNA, RNA,
or proteins Viroids
- Antibiotics function by inhibiting certain - consist of short, naked fragments of
metabolic activities within cellular single-stranded RNA (about 300-400
pathogens. It may be prescribed in an nucleotides in length) that can interfere with
attempt to prevent secondary bacterial the metabolism of plant cells and stunt the
infections that might follow virus infection. growth of plants, sometimes killing the
plants in the process
Oncogenic Viruses - Transmitted between plants in the same
- Viruses that cause cancer manner as viruses
- Epstein-Barr Virus (nasopharyngeal
carcinoma, Burkitt lymphoma, and B-cell Potato spindle tuber
lmphoma)
- Human herpes virus (Kaposi sarcoma) Prions
- small infectious proteins that apparently
Human Immunodeficiency Virus (HIV) cause fatal neurological diseases in animals
- Enveloped, single stranded RNA virus - Believed to be the most resistant to
- From genius Lentiviruses, family disinfectants
Retroviridae (retroviruses) - Stanley B. Prusiner - studied the role of
- Able to attach to and invade cells bearing these proteinaceous infectious particles in
receptors (CD4) that virus recognizes disease)

Mimivirus (Mimics) Mad cow disease

- Extremely large double-stranded DNA
virus (7 nm thick capsid with a diameter of
750 nm)
- Recovered from Amoebas Domain Bacteria Identification and Classification
- It “mimics” the bacteria
• 23 phyla • Stain reaction
- Observable using standard compound light • 32 classes • Cell shape and morphological arrangement
microscope • 5 subclasses • Motility
• 77 orders • Colony morphology
- Some of its genes code for translation of • 14 suborders • Atmospheric and nutritional requirements
protein and DNA repair enzymes • 182 families • Biochemical and metabolic activities
• 871 genera • Specific enzymes that the organism
- Contains several genes for sugar, lipid, • 5,007 species produces
• Pathogenicity
and amino acid metabolism
- Reported to cause some cases of Human
pneumonia

Plant Viruses
- more than 1,000 different viruses cause
plant disease resulting in huge economic
losses

Graciel Aronce
BSM 2-3
 DOMAIN BACTERIA TREE PHENOTYPE Arrange Descripti Appeara Example Disease
ment on nce
CATEGORIES
Diplococci Cocci in Neisseria Gonorrhe
• Reproduce by Binary • Gram-positive with pairs
gonorrhea
a
Fission cell wall
• After binary fission, the • Gram-negative with Streptococc Cocci in Streptococcus
pyogenes
Strep
i chains throat
daughter cells may separate cell wall
completely from each other • Lacks cell wall
Staphyloco Cocci in Staphylococc Boils
or may remain connected, cci us aureus
clusters
forming various morphologic
arrangements. Tetrad A packet Micrococcus Rarely
luteus
of 4 cocci pathogeni
Organism’s generation c
time - The time it takes for
one cell to split into two Octad A packet Sarcina Rarely
of 8 cocci ventriculi pathogeni
cells. c

CELL MORPHOLOGY
• Cocci
• Bacilli
• Spirilla

3 BASIC SHAPES OF BACTERIA

Bacterial shape and morphological

arrangement

2 Bacilli (Rectangular or Rod-shaped)

1 Cocci (Round or Spherical)
Morphological Arrangement
Morphological Arrangement • diplobacilli = pair
• diplococci = pair • streptobacilli = chain
• streptococci = chain • coccobacilli = short rods/ovals
• staphylococci = clusters
• tetrads = pack of 4 E.g. Haemophilus influenzae
• octads/sarcina = pack of 8

Graciel Aronce
BSM 2-3
 STAINING PROCEDURES

STAINING PROCEDURES - utilized to

examine bacteria
• Smear - bacterial samples are smeared on
to glass microscope slide
• Air-dry
- important to ensure that there is no excess
water from the smear and that smear is
Spirilla (Curved and spiral-shaped
completely dry before heat fixing
bacilli) - rigid and flexible
- helps the bacterial sample to adhere
properly to the slide preserving the
Morphological Arrangement
morphology and structural integrity of the
• Curved (comma-shaped)
cell
(Vibrio spp. (species))
• Fix
- Cholera
Subdivided into two:
• gull-wing (resembles a bird)
> Heat fixation - passing the smear through
(Campylobacter spp.)
a Bunsen burner flame
- Diarrhea
- during heat fixing, any remaining water
• Spirochetes - helical and flexible
could boil which potentially causes cell
> Tightly coiled
rupture and altering cellular morphology
(Treponema pallidum)
- excess heat can destroy the morphology
- Syphilis
of the cell
> Less tightly coiled
(Borrelia spp.)
> Methanol fixation
- Lyme disease
- flooding the smear with absolute
methanol for 30 secs
- a more satisfactory fixation technique
Cell Wall-Deficient (CWD) Bacteria
• Lose their characteristic shape due to
Fixation serves 3 purposes:
adverse growth conditions preventing the
• It kills the organisms.
production of normal cell walls
- ensures that the samples are no
• Some may revert to their original shape
longer viable (Capable of surviving
when placed in favorable growth conditions
or living successfully) reducing the
risks of contamination or infection
Genus Mycoplasma
• It preserves the morphology (shape)
• No cell wall
- important for accurate identification
• Resistant to antibiotics
and observation of the sample
• It anchors the smear to the slide
Pleomorphism - the ability to exist in a
variety of shapes

Graciel Aronce
BSM 2-3
 - crucial part of preventing the smear DIFFERENTIAL STAINING PROCEDURES
from being wash away during the
staining procedure 1 Gram Staining
Dr. Hans Christian Gram while working in a
laboratory in Berlin Hospital developed a staining technique
STAINING TECHNIQUES that would enable him to see bacteria in the lung tissues of
Specific stains and staining techniques are patients who had died of pneumonia
utilized to observe bacterial cell morphology - Developed in 1883 (Observation of Bacteria in
a patient’s lung tissue die of pneumonia)
- The most important staining procedure in
• SIMPLE STAINING PROCEDURE bacteriology laboratory, as it differentiates the
- sufficient to determine bacterial shape and gram + and the gram - bacteria:
morphologic arrangement
- uses single stain solution The organism’s gram reaction serves as the
important clue when attempting to learn the
Simple Bacterial Staining Technique identity of a particular bacterium. The color of
• With a flamed loop, smear a loopful of bacteria the bacteria at the end of the gram staining
suspended in broth or water onto a slide procedure depends on the chemical composition
• Allow slide to air-dry of their cell wall:
• Fix the smear with absolute (100%) methanol Blue to Purple: If the bacteria will not
• Flood the slide with the stain (Methylene blue) decolorized during the decolorization step
Methylene blue (or Methylthioninium
chloride) Pink to Red: If the crystal violet is removed from
- applied to the fixed smear the cells during decolorization step and the cells
- positively charged basic tighted binds to the are subsequently stained with safranin (red dye)
negatively-charged component of the
bacterial cell resulting in a blue stain
Gram-positive (blue to purple stain)
- enhances the visibility of bacterial cells and
reveals structural details facilitating improved - thick layer of peptidoglycan in the cell
observation of morphology and arrangement walls of the bacteria makes it difficult to
• Rinse with water and blot dry with bibulous remove the crystal violet iodine complex
paper or paper towel. during the decolorization step
- After adding the stain, the sample will be -
rinsed with water to remove excess dye, Gram-negative (pink to red stain)
enhancing the contrast between the stained - thin layer of peptidoglycan in the cell
cells and the background allowing for a walls
clearer visualization of cellular structures and
the morphology
Peptidoglycan - mainly a polysaccharide
- Sample is then dried to fix the dye within the
composed of two chemical subunits found only
cell and examine under the microscope
in the bacterial cell wall.
• Examine the slide with 100x microscope
objective, using a drop of immersion oil directly
🟣
Legend:
on the smear
🔴
- Gram POSITIVE
- Gram NEGATIVE
COLORLESS (decolorized) - Gram NEGATIVE
STRUCTURAL STAINING PROCEDURE
- Used to observe bacterial capsules, spores,
and flagella

Graciel Aronce
BSM 2-3
 DIFFERENCES BETWEEN GRAM-POSITIVE AND
- Heat is necessary because the cell walls
GRAM-NEGATIVE BACTERIA of mycobacteria contain pax which
prevents the stain from penetrating the
Gram- Positive Gram-Negative
Bacteria Bacteria cell. The heat softens the wax and able
to , a decolorizing agent which is a
Color at the end Blue to purple Pink to red mixture of acid and alcohol is then used
of the Gram
Staining in an attempt to remove red color from
Procedure the cell. Most other bacteria are
decolorized by the acid-alcohol
Peptidoglycan in Thick layer Thin layer
cell walls treatment and are said to be non-acid
fast.
Teichoic acids Present Absent
and lipoteichoic
acids in cell walls - Commonly utilized for Mycobacterium spp.
(species) classified as acid-fast bacteria due to the
Lipoposaccharid Absent Present
resistance to conventional staining with aniline dye
e in cell walls
such as the gram stains & their ability to retain dye
even after treatment with acid alcohol decolorizers.
This resistance is attributed to the unique
composition of their cell wall, which contains high
Examples: level of glycolic acids forming a thick lipid-rich and
hydrophobic barrier that … the cell wall impermeable
to gram staining. As a result mycobacterium either
Gram-Positive Bacilli
remains unstained or exhibit inconsistent staining
(Clostridium tetani) - causative agent of tetanus
results.

- Useful for Tuberculosis laboratory due to its

Loosely-coiled Gram-Negative Spirochetes specificity to mycobacterium tuberculosis
(Borrelia burgdorferi) - causes lyme disease (causative agent of tuberculosis)
transmitted by an infected tick > acid-fast bacilli (AFB) in sputum specimen

CHARACTERISTICS OF SOME IMPORTANT

2 Acid-Fast Stain PATHOGENIC BACTERIA
Paul Ehlrich - pioneered the
- Developed in 1882
STAINING MORPHOL BACTERIU DISEASE(S
- Carbol fuchsin (Ziehl-Neelsen) stain + Heat REACTION OGY M )
> a basic bright red dye is used for
bacterial staining procedure Gram-positi Cocci in Staphyloco Wound
ve clusters ccus aureus infections,
> Introduced by Franz Ziehl as a primary bollus,
dye and further enhanced by Friedrich pneumonia
Cocci in Streptococc
Neelsen by incorporating heat to chains us Strep
facilitate dye penetration pyogenes throat,
> developed the modern Ziehl-Neelsen scarlet
Diplococci fever
in 1882
- This staining technique involves initial
application of carbol fuchsin bacterial
smear and then applying heat by
passing a Bunsen burner flame beneath
the slide until the carbol fuchsin sticks

Graciel Aronce
BSM 2-3
 - If organism is a
Endospore spore-former
stains (terminal or
subterminal
spores)

Differential Gram stain - Gram-positive

vs.
Gram-negative

Acid-fast - Acid-fast or
stain Non acid-fast

MOTILITY

- ability to “swim”
- often associated with the presence of flagella
or axial filaments

Monotrichous (Mono: One)

- possess single flagellum at one end of the cell

SUMMARY OF BACTERIAL STAINING
PROCEDURES Lophotrichous (Lopho: Crested at one end)

- multiple flagella clustered at one end of the cell

Amphitrichous (Amphi: on both sides)

- bacteria that has a single flagellum at both

ends

Peritrichous (Peri: around the surface)

- bacteria that have flagella distributed all over

the surface of a cell
Category Example Purpose

Simple Staining with To determine

methylene size, shape, and TWO DIFFERENT WAYS IN
blue morphology DEMONSTRATING BACTERIAL MOTILITY
Structural Capsule - Whether the
stains organism is 1 STABBING BACTERIA INTO A TUBE OF
encapsulated SEMISOLID AGAR

- If organism
Flagella possesses
stains flagella & its no.
and loc

Graciel Aronce
BSM 2-3
 • Microaerophilic aerobes
- require reduced oxygen concentrations
(usually around 5% oxygen)

• Obligate anaerobes
- do NOT require oxygen for life and
reproduction
- Growth of bacteria in a semisolid agar results
in turbidity or cloudiness. • Aerotolerant anaerobes
Non-motile organisms - only exhibit growth - do NOT require oxygen, grows better in the
along the stab line resulting in turbidity (measure absence of oxygen, but can survive in
of relative clarity of a liquid) restricted to the line. atmospheres containing molecular oxygen.

Motile organisms - spread from the stab line • Facultative anaerobes

producing turbidity throughout the medium. - can thrive in an atmosphere with or devoid of
oxygen, any where from 0% O2 to 20% to 21%
2 HANGING DROP TECHNIQUE O2

Easy Way to Remember: “OF FAME”

This acronym helps recall each category’s oxygen requirement:

● O: Obligate Aerobes – Oxygen-only

● F: Facultative Anaerobes – Flexible with oxygen
- A drop of bacterial suspension is placed on a glass ● F: Aerotolerant Anaerobes – Fine without oxygen, not
affected by it
cover slip which is then inverted over a depression ● A: Obligate Anaerobes – Avoid oxygen
slide, upon microscopic examination: ● M: Microaerophiles – Minimal oxygen needed
Motile bacteria within the hanging drop moving in ● E: Environment placement – Remember where they grow
in liquid culture:
various directions
○ Top: Obligate Aerobes
○ Top and throughout: Facultative Anaerobes
COLONY MORPHOLOGY (appearance) ○ Evenly: Aerotolerant Anaerobes
○ Bottom: Obligate Anaerobes
A mound or pile of bacteria (bacterial colony) ○ Just below the top: Microaerophiles
- Includes the size, color, overall shape,
elevation, and the appearance of the edge or the
margin of the colony.
NUTRITIONAL REQUIREMENTS
• Elements
Size of colonies is determined by the organism’s
- carbon, hydrogen, oxygen, sulfur, phosphorus,
rate of growth (generation time)
and nitrogen
- potassium, calcium, iron, manganese,
magnesium, cobalt, copper, zinc, and uranium
ATMOSPHERIC REQUIREMENTS
• Vitamin requirements
- classification of bacteria based on their
• Organic substances
relationships to oxygen (O2) and carbon dioxide
(CO2)

• Obligate aerobes
- require an atmosphere containing about 20%
to 21% oxygen

Graciel Aronce
BSM 2-3
Easy Way to Remember: “C H E M O G R • Tested by injecting the organism into mice or
A P H” cell cultures.

This acronym highlights the main components bacteria need to grow: Genetic Composition
- molecular diagnostic procedures
● C – Carbon source
● H – Hydrogen and Water (H₂O) ● DNA probes
● E – Energy source (Light for Phototrophs, Chemicals for ● 16S rRNA sequencing
Chemotrophs)
● M – Minerals (e.g., iron, potassium, magnesium)
● O – Oxygen requirement (depending on the bacteria’s Unique Bacteria
type)
• Rudimentary Bacteria
● G – Growth factors (like vitamins and amino acids)
● R – Reduced Nitrogen (for making proteins and nucleic • Formerly thought to be a virus
acids) - Rickettsias, chlamydias, and
● A – Autotrophs and Heterotrophs (types based on carbon
source) mycoplasmas
● P – Phosphorus (for ATP and nucleic acids)
● H – Helpful elements (trace elements and cofactors)
• Rickettsia and Chlamydia
Mnemonic Summary: - Gram-negative
Use "CHEMOGRAPH to remember the essentials for bacterial - Obligate intracellular pathogens
growth: Carbon, Hydrogen, Energy, Minerals, Oxygen, Growth
factors, Reduced nitrogen, Autotroph/Heterotroph, Phosphorus,
Helpful elements." RICKETTSIA
Howard T. Ricketts

Fastidious organisms
• Leaky cell membranes
• Especially demanding nutritional requirements
• Must live inside another cell to retain all
• Special enriched media
necessary cellular substances
• Arthropod-borne disease
- Lice, fleas, and ticks
BIOCHEMICAL & METABOLIC ACTIVITIES
• Rash-producing disease
• Bacteria growth led to production of waste
products and enzyme secretions
CHLAMYDIA
• In particular environments, some bacteria are
Ludwig Halberstädter and Stanislaus von
characterized by the production of certain
Prowazek
gasses (e.g. CO2, H2S, O2, CH4)
• Identification of certain types of bacteria in the
• Energy parasites
laboratory are done by inoculation into various
- Able to produce adenosine triphosphate
substrates
(ATP) molecules
• Determine whether they possess the enzymes
• Obligate intracellular pathogens
necessary to break down those substrates.
- Aerosis or by direct contact between
hosts
Group B Streptococcus (GBS)

MYCOPLASMA
PATHOGENICITY
Louis Dienes and Geoffrey Edsall
- characteristics that enable bacteria to cause
disease
• Smallest of the cellular microbes
• Pleomorphic
• Capsules, pili, or endotoxins (Gram-negative
• Pleuropneumonia-like organisms
bacteria)
• Free-living or parasitic
• Secretion of exotoxins and exoenzymes
• Resistant to treatment with penicillin and other
antibiotics

Graciel Aronce
BSM 2-3
• Can be cultured on artificially media - Purple and Green bacteria
- fried egg colonies

CYANOBACTERIA
HUMAN DISEASES CAUSED BY UNIQUE
• Site of photosynthesis
BACTERIA
- Thylakoids
• Phycobilisomes
ESPECIALLY LARGE BACTERIA
- Complex protein pigment aggregates
• Believed to be the first organism capable of carrying out oxyge
Thiomargarita namibiensis
photosynthesis
- colorless, marine, sulfide-oxidizing bacterium
- Fossil records reveal that cyanobacteria already in existe
- 100 to 300 micrometer, but may be as large as
3.3 to 3.5 billion years ago
750 micrometer (0.75 mm)
• Vary widely in shape
• Water bloom (pond scum)
Epulopiscium fishelsoni
- mild or no wind
- 80 micrometer wide, 600 micrometer (0.6 mm)
- balmy water temp (15º - 30ºC)
long
- pH of 6-9
- abundance of the nutrients nitrogen & phosphorus
Metabacteria
• Nitrogen fixation
- 20 to 30 micrometer in length

ESPECIALL SMALL BACTERIA

Nanobacteria TAXONOMIC DOMAIN ARCHAEA

- 20 nm in diameter CLASSIFICATION
- Found in soil, minerals, ocean water, human • Archaebacteria
and animal blood, human dental calculus • 2 phyla (Archaeobacteria)
• 8 classes • Ancient
(plaque), arterial plaque, and even rocks
• 12 orders • Closely related to
(meteorites) of extraterrestrial origin Eukaryotes
• 2 families
- Some scientists believe that these tiny • Vary widely in shape
• 69 genera
structures were formed by geological, rather • 217 species • Extremophiles
than biological processes. - Methanogens
• Possess cell walls
- No
peptidoglycan
PHOTOSYNTHETIC BACTERIA
• Purple bacteria, Green bacteria, and
Cyanobacteria (Blue green algae)
• Some may revert to their original shape when
placed in favorable growth conditions EXAMPLES OF EXTREMOPHILES
• Capable of converting light energy into
chemical energy Type of Extreme Name given to these
Environment Types of
Oxygenic photosynthesis Extremophiles
• Photosynthesis that produces oxygen
- Cyanobacteria Extremely acidic Acidophiles

Extremely alkaline Alkaliphiles

Anoxygenic photosynthesis
• Photosynthesis that does not produce oxygen

Graciel Aronce
BSM 2-3
 Extremely hot Thermophiles dioxide as
an energy
Extremely cold Psychrophiles source

Extremely salty Halophiles Store energy Yes Yes

in the form
Extremely high Piezophiles (formerly of starch
pressure barophiles)
Compose of No Most
roots, (bryophytes,
stems, and such as
leaves mosses, are
the
exception)
MICROBIAL DIVERSITY:
EUKARYOTIC MICROBES Cell walls Most Yes
contain (exceptions
cellulose include
diatoms and
ALGAE (plural) / ALGAL (singular) dinoflagellate
- Photosynthetic, eukaryotic organisms s; Euglena
- referred to as protist and Volvox
- Phycology (or Algology) do no have
cell walls)
CHARACTERISTICS AND CLASSIFICATION
Method of Both asexual Sexual
• Cell consist of reproductio and sexual
- cytoplasm, a cell wall (usually), a cell n
membrane, a nucleus, plastids,
ribosomes, mitochondria, and Golgi Contain a No Most
bodies vascular (mosses and
- Pellicle system to other
transport bryophytes
- Stigma (or Eyespot) - light sensing
internal are
- Flagella fluids avascular)
- Lack true roots, stems, and leaves

CHARACTERISTICS AND CLASSIFICATION

Similarities and Differences between Algae • Not all algae are microorganisms
and Plants • May be arranged in colonies or strands
• Produce energy by Photosynthesis
ALGAE PLANTS - Few species use organic nutrients, and
others survive with very little sunlight
Eukaryotic Yes Yes
• Most algal cell walls contain cellulose
Photosynthe Yes Yes
tic Classification based on the type of
photosynthetic pigments:
Cells Yes Yes • Green
contain
• Golden (or Golden Brown)
chlorophyll
• Brown
Use carbon Yes Yes • Red

Graciel Aronce
BSM 2-3
 (In the absence of light, euglena can obtain
MICROSCOPIC ALGAE nutrients by ngesting food through cyostome )
- cystostome
• Diatoms - Lack of cell wall but has pellicle (for protection)
- Tiny, usually unicellular algae that live in both - stigma and a single flagellum
freshwater and seawater
- Important members of Phytoplankton MACROSCOPIC ALGAE
- Cell walls contain Silicone Dioxide (SiO2)
giving a glass-like structure • Brown Kelp
- Diatomaceous earth - produced by deposits of - up to 10 m in length
diatoms which is applied in filtration system, - found along ocean shores
insulations
• Green scum
• Dinoflagellates - found floating on ponds
- Microscopic, unicellular, flagellated, often
photosynthetic algae • Slippery green
- Important members of the phytoplankton - found on wet rock
- Producing most of our atmospheric oxygen and
serving as key links in food chains
- Fire algae
- Responsible for Red tides (harmful algal
blooms or HAB which gives the water reddish
green produced toxins that can be hardly
dangerous to many organsms)

MICROSCOPIC ALGAE (Green Algae)

• Desmids
- unicellular algae

• Spirogyra
- filamentous alga
ALGAE
• Chlamydomonas Medical Significance
- unicellular, biflagellated alga
- containing one chloroplast and a stigma Genus Prototheca
• Enter wounds especially those located in feet
• Volvox - Protothecosis
- Multicellular alga, often referred to as colonial - small subcutaneous lesion
alga or colony - crusty, warty looking lesion
- 60,000 interconnected, biflagellated cells, • Enters the lymphatic system
arranged to form a hollow sphere - Debilitating, sometimes fatal infection,
- move through the water in rolling motion due to especially in immunosuppressed individuals
coordinated dating of their flagella • Phycotoxins

• Euglena
- Possesses both features of algae and protozoa PROTOZOA
- Contains chloroplasts, is photosynthetic, and • Eukaryotic organisms
stores energy in the form of starch • referred to as protist

Graciel Aronce
BSM 2-3
• Protozoology Protozoas
Subdivided based to their Method of
CHARACTERISTICS AND CLASSIFICATION Locomotion
• Unicellular, ranging in length from 3 to 2,000
micrometer • Amebae (Amoeboid Protozoans)
- Free-living organisms, found in soil and water - Pseudopodia or “false feet” - cytoplasmic
- Protozoal cells are more animal-like extensions
• Cells consists of: - Ameboid movement
- Cell membranes, nuclei, endoplasmic - phagosomes
reticulum, mitochondria, Golgi bodies, - Phagocytes
lysosomes, centrioles, and food vacuoles - Pinocytosis
• Some possess pellicle, cytostomes, contractile
vacuoles, pseudopodia, cilia, and flagella. • Ciliate (Ciliated)
• No chlorophyll - Cilia
• Ingest whole algae, yeasts, bacteria, and - Oar-like motion
smaller protozoans as their source of nutrients - Most complex of all protozoa
- Others live on dead and decaying organic - Blepharisma, Didinium, Euplotes,
matter Paramecium, Stentor, and Vorticella spp.
• No cell wall
• Some flagellates and some ciliates: • Flagellate (Flagellated)
- Pellicle (protection) & Cytostome (food - whip-like flagella
ingestion) - Kinetosome (or Kinetoplast)
Pellicle - a substitute of protozoan for not - Wave-like motion
having a cell wall which is a thickened cell - some are known to be pathogenic
membrane
• Some pond water protozoa (amebae & • Non-motile (Sporozoans)
paramecium) - sporozoa - lacks pseudopodia, flagella, or cilia
- Contractile vacuole - Plasmodium spp.
• Vorticella spp. (pond water ciliates) - Malaria
- contractile stalk (myoneme)
• Parasitic protozoa
- Break down and absorb nutrients from the
body of the host in which they live
- Pathogens
- Trichomoniasis
• Mutualistic Symbiotic relationship
- E. coli is one of the first bacteria to colonize
neonates at birth (Human gut)

PROTOZOAN LIFE CYCLE

PROTOZOA
• Cyst Stage
Medical Significance
- nonmotile, dormant, survival stage
- like bacterial spores
Entamoeba histolytica
• Amoebiasis
• Trophozoite Stage
- the motile, feeding, and dividing stage
Naegleria fowleri

Graciel Aronce
BSM 2-3
• primary amebic meningoencephalitis - Asexual spores
Balantidium coli • Some species of fungi produce both asexual
• Dysentery in underdeveloped countries and sexual spores
• Fungal spores are very resistant structures that
Trichomonas vaginalis (STI) are carried great distances by wind
• Tricomoniasis - Heat, cold, acids, bases, and other chemicals
- Allergen
Cryptosporidium parvum
• Cryptosporidiosis PHYLA OF FUNGI

Zygomycotina (Zygomycota)
FUNGI • Common bread molds
• Kingdom fungi • Fungi causing food spoilage
• Mycology - encompasses the study of fungi
Chytridiomycotina (Chytridiomycota)
CHARACTERISTICS AND CLASSIFICATION • Water molds
• The saprophytic fungi - Lives in water and soil
- Living on organic matter in water and soil • Not considered to be true fungi by taxonomists
• The parasitic fungi
- Living on and within animals and plants Ascomycotina (Ascomycota)
• Live on many unlikely materials • certain yeasts and some fungi that cause plant
- Deterioration of leather and plastics and disease
spoilage of foods
- Essential in the production of cheeses, Basidiomycotina (Basidiomycota)
alcoholic beverages, and other foods, as well as • some yeasts, some fungi that cause plant
certain drugs & antibiotics diseases, and the large “fleshy fungi” live in the
• Include yeasts, molds, and mushrooms woods
• Saprophytes - mushrooms, toadstools, bracket fungi,
- Secretes digestive enzymes and decompose puffballs
material into absorbable nutrients
• Not photosynthetic Deuteromycotina (Deuteromycota)
• Cell wall contains chitin • Fungi imperfecti
• Hyphae • Contains fungi having no mode of sexual
- Mycelium (Thallus) reproduction
• Septate hyphae • Important molds
- cytoplasm within the hypha is divided into - Aspergillus and Penicillium
cells by septa
• Aseptate hyphae
- cytoplasm within the hypha is not divided into
cells YEAST
- Coenocytic - unicleated cytoplasm • Unicellular organisms
• Lack mycelia
REPRODUCTION • Blastospores (blastoconidia)
• Budding • Reproduced by budding
• Hyphal extension - occasionally do so by a type of spore
• Formation of spores formation
- Sexual spores • Pseudohypha
- produced by the fusion of two gametes • Chlamydospores (chlamydoconidia)
- ascospores, basidiospores, zygospores

Graciel Aronce
BSM 2-3
• Found in soil and water and on the skins of Key Differences Between Prokaryotes and
many fruits and vegetables Eukaryotes
• Louis Pasteur
- Discovered naturally occurring yeasts on the Feature Prokaryotes Eukaryotes
skin of grapes and other fruits and grains were
Nucleus No true True nucleus
responsible for fermentation processes
nucleus with nuclear
(DNA is membrane
free-floating)
SUMMARY:
Size Smaller (1-10 Larger
micrometers) (10-100
Prokaryotic Microbes:
micrometers)
● Definition: Simple, single-celled
Organelles No Membrane-b
organisms without a nucleus or other
membrane-b ound
membrane-bound organelles.
ound organelles
● Main Characteristics: DNA is
organelles (mitochondria
free-floating within the cell, not enclosed
, ER)
within a nucleus.
● Examples: Bacteria (like E. coli) and DNA Circular DNA Linear DNA
Archaea (like methanogens). Structure in
● Key Points: Prokaryotes are ancient chromosome
and structurally simpler than eukaryotes. s

Eukaryotic Microbes: Reproductio Asexual Both asexual

n (binary and sexual
● Definition: More complex cells with a fission) (mitosis and
nucleus that houses the cell’s DNA, and meiosis)
they have membrane-bound organelles
(e.g., mitochondria). Examples E. coli, Amoeba,
● Main Characteristics: DNA is enclosed Archaea yeast, algae
within a nuclear membrane, and the cell
contains organelles with specialized
functions. Examples of Each Type
● Examples: Protozoa (like Amoeba),
fungi (like yeast), and algae. ● Prokaryotic Microbes:
● Key Points: Eukaryotes can be ○ Bacteria: Single-celled
single-celled or multicellular, and they’re organisms, often studied in
generally larger and more complex than microbiology. Example:
prokaryotes. Staphylococcus aureus (causes
skin infections).
○ Archaea: Single-celled, often
extremophiles. Example:
Methanogens (produce
methane in anaerobic
environments).

Graciel Aronce
BSM 2-3
 ● Eukaryotic Microbes:
○ Protozoa: Single-celled
organisms, often found in water.
Example: Paramecium (moves
using cilia).
○ Fungi: Includes yeasts and
molds. Example:
Saccharomyces cerevisiae
(brewer’s yeast).
○ Algae: Photosynthetic
organisms found in water.
Example: Chlamydomonas (a
green alga).

Easy Way to Remember: “PRO vs.

EU”

Using the mnemonic “PRO-NO; EU-DO” helps

to remember the main differences:

● PRO-NO: Prokaryotes have No

nucleus and No organelles.
● EU-DO: Eukaryotes Do have a
nucleus and Do have organelles.

Recap Summary

● Prokaryotic Microbes: Simple,

single-celled, no nucleus, small, and
reproduce asexually. Examples:
Bacteria and Archaea.
● Eukaryotic Microbes: Complex, have a
nucleus and organelles, larger, and
reproduce both sexually and asexually.
Examples: Protozoa, fungi, algae.

Graciel Aronce
BSM 2-3