I.

INTRODUCTION

Microbiology
- study of all living organisms that are too small to be seen by the naked eye
- Latin “mikros” small “bios” life, “logia” study of
- ultimately, the methods used on how to manipulate these minute organisms and how they
can affect our daily lives.

II. BRANCHES OF MICROBIOLOGY

● By Taxonomy (BINMVPPP)
○ Bacteriology- study of bacteria
○ Immunology- study of immune system: relationship between their pathogens
(bacteria and virus) to their host.
○ Nematology- study of nematodes (roundworms)
○ Mycology- deals with various fungal microorganisms (fungi, yeast, mold)
○ Virology- involves multicellular and unicellular virus
- only become living when host attaches
○ Phycology- study of algae
-single cell organisms like amoeba
○ Protozoology- study of protozoa
○ Parasitology- study of parasites

● By Type of Applied Microbiological Research (FAMMP)

○ Food Microbiology-study of microbes that spoil food and cause food-borne
illnesses. It is also the study of food production(beer fermentation)
○ Agricultural Microbiology- study of microorganisms that interact with plants and
soil
○ Microbial Biotechnology- involves microbes in industrial and consumer products
○ Medical Microbiology- study of microorganisms responsible for human disease
○ Pharmaceutical Microbiology- study of microorganisms used in pharmaceutical
products such as vaccines and antibiotics

III. RELEVANCE TO NURSING

● Proper specimen handling - avoid contamination and spread of disease
● Strict compliance to sanitation procedures- Prevent spread of infection
● Vaccination- Prevent spread of infection
● Proper patient management- proper administration of medicine

IV. HISTORY OF MICROBIOLOGY (ARLF RJJ LS)

● Anton Van Leeuwenhoek
- Father of Microbiology
- First to publish observations about bacteria by looking at water with a microscope named it
“ANIMALCULES” (now called animal and plant cells)
- Also discovered spermatozoa, erythrocytes, single celled algae and ciliates
-
● Robert Koch
- “Father of Modern Bacteriology”
- Discovered anthrax disease cycle (contracted through contaminated meat or animals,
leads to sepsis, hemorrhagic meningitis and damage to multiple organs in the body)
- Discovered bacteria for tuberculosis and cholera
 - Discovered tuberculin skin test
- Germ Theory

● Louis Pasteur
- French chemist and microbiologist
- Discovered principles of Vaccination(for anthrax, cholera and rabies), Microbial
Fermentation (all fermentation caused by microorganism like how yeast ferments alcohol
and sugar) and Pasteurization
- Ended debate with Theory of Spontaneous Generation by boiling meat broth in a swan
neck flask (tindog-clear; higda; cloudy)
-
● Francisco Redi
- “Father of Modern Parasitology”
- Theory of Biogenesis
- states that all complex living things comes from living things by reproduction
- Life does not arise from non living material
- This is done by experimentation on 3 jars:
1. Open jar with fresh meat; infested with maggots after few days
2. Sealed jar filled with fresh meat; no signs of maggots
3. Jar covered with gauze pad; maggots stayed on top of gauze pad
-
● Robert Hooke
- First used the word “CELL” by viewing microscopic cavities of cork
-
● Jan Baptiste Evan Helmont
- Known for his support and additional information to Theory of Spontaneous Generation
- He experimented using mice and wheat kernels in racks for 3 weeks; he noticed na
nidaghan daw ang mice pero delulu ra diay siya CHAR!
-
● John Needham
- He agreed with Theory of Spontaneous Generation by boiling broth infused with animal
and plant matter; hoping to kill all microorganisms
- Observed that broth had become cloudy and single drop contained numerous microscopic
creatures
-
● Lazzaro Spallanzani
- He disagreed with John Needham by doing the same experiment but this time, boiled
broth in a prolonged time
- He agreed with Theory of Spontaneous Generation but experiment results were
contradictory
-
● Shibasaburo Kitasato
- Discovered bacterium that causes the bubonic plague Pasteurella pestis, but now
Yersinia pestis together with Alexander Yersin (by dealing with cadavers of rats, he
examined lymph nodes)
- Demonstrated importance of antitoxin to prevent diseases such as diphtheria and tetanus
- Discovered serum therapy together with Von Behring; injected susceptible animals like
dog and cats; antitoxins from blood of animals that were exposed to tetanus.

Other Notes:

● Theory of Spontaneous Generation was first made by Aristotle. It states that living things
can arise from nonliving things
 V. HISTORY OF CLASSIFICATION OF ORGANISMS

Taxonomy- systematic classification of living organisms

TAXONOMISTS:

● Aristotle
- classified living things as plants and animals
● Carl Linnaeus
- introduced the classification system thru systemic method (taxonomic
nomenclature), giving the organism a genus and species
● Ernst Henrich Haeckel-
- 3 different categories according to function:
1. Animalia (Animals)
2. Plantae (Plants)
3. Protista (Eukaryotes)
● Carl Woese
- 3 domains system/kingdoms:
1. Archaebacteria
2. Eubacteria
3. Eukaryotes
a. Protista
b. Fungi
c. Animalia
d. Plantae

Prokaryotes Similarities Eukaryotes

● Unicellular ● Have DNA ● Unicellular or multicellular
● Absent nucleus ● Have ● nucleus is present
● Circular DNA ribosomes ● linear DNA
● lack membrane ● Have ● has membrane
● bound organelles cytoplasm ● bound organelles
● reproduce both sexually ● Have ● cell division by mitosis
and asexually glycocalyx ● examples are plant and animal
● cell division by binary ● Have cell cells,including humans
fission membrane
● examples are bacteria and
archaea celis
 CHARACTERISTICS PROKARYOTES EUKARYOTES

1. Nucleus No: Nucleoid only Yes

2. Membrane Bound No Yes

Organelles

3. Cell Wall containing Yes No

Peptidoglycan

4. Plasma Membrane Yes Yes

5. Cytoplasm No?? Yes

6. Ribosomes Yes but smaller in size Yes but larger

7. Cell Division Binary Fission Mitotic Division

8. Chromosomes Single , Short-Lived Multiple, Long lived, Processed

Unprocessed RNA mRNA

9. Sexual Reproduction No Meiosis Yes: Meiosis

10. Glycocalyx Yes: AS A CAPSULE OR SLIME Yes: COMPOSED OF CHO;

LAYER/ REGULATES STRENGTHEN CELL SURFACE
PERMEABILITY OF ; signal reception between cells
NUTRIENTS;provides a and the environment.
protective coat from host factors

Common bacteria and their associated infection/disease

● Mycobacterium tuberculosis- Causative agent of Tuberculosis

● Vibrio cholerae- Causative agent of Cholera
● Corynebacterium diphtheriae- causative agent of diphtheria
● Treponema pallidum- Causative agent of Syphilis
● Clostridium tetani- Agent of tetanus
● Bacillus anthracis- Causative agent of anthrax disease
● Escherichia coli- Most common cause of UTI.
● Chlamydia trachomatis- Causative agent of chlamydia.
● Mycobacterium leprae- Causative agent of leprosy
● Salmonella typhi- Causative agent of typhoid fever.
● Candida albicans- Causative agent of oral/ genital thrush.
● Yersinia pestis - Causative agent of plague.
● Bacillus cereus- Bacteria associated with food-poisoning.
● Neisseria gonorrheae- Causative agent of an STI, gonorrhea
● Streptococcus pneumoniae- Bacteria causing Community Acquired Pneumonia(CAP)
● Methicillin Resistant Staphylococcus aureus (MRSa)- Leading cause of Hospital
Acquired Pneumonia(HAP)
● Staphylococcus epidermidis - Most frequent contaminant of blood units.
● Streptococcus pyogenes- Common bacteria causing sore throat and tonsils.
 VI. FIVE BASIC GROUPS OF MICROORGANISMS
1. Prokaryotes (Bacteria)
2. Eukaryotes (Fungi, Mold, Yeast)
3. Protozoan (Parasites)
4. Metazoan (Helminths)
5. Virus

VII. CRITERIAS USED IN CLASSIFYING

1. Morphology
2. Locomotor structures
3. Colony Characterestics
4. Metabolism
5. Staining Characteristics
6. Endospores
7. Nutrition
8. Growth Characteristic
9. Genetic Characteristics

VIII. BACTERIAL CELL ANATOMY

● Layers Outside the Cell Wall

○ Flagella
○ Capsule
○ Pili/Fimbriae

●
Layers within the Cytoplasm
○ Cytoplasm/Protoplasm
○ Nucleoid
○ Ribosomes
○ Endospore
○ Inclusion Granules
○ Cytoplasmic Membrane
OUTSIDE
A. CAPSULE
- is a polysaccharide layer that completely envelopes the cell.
- well organized and tightly packed, which explains its resistance to staining under the
microscope.
- Offers protection from a variety of threats to the cell.
- enhance the ability of bacterial pathogens to cause disease and can provide
protection from phagocytosis
- It can help in attachment to surfaces and determines the virulence factor of
organisms.
Disease Causing Organisms
•S. Pneumoniae – capsulated
•B. anthracis – capsule of pure D- glutamic acid
•Y. pestis – capsules of mixed amino acids.
 B. APPENDAGES
a. FLAGELLA
- protein appendages for locomotion (Movement of bacteria)
- consists of a basal body, hook, and a long filament composed of a
polymerized protein called flagellin
- 6 KINDS
1. Atrichous- no flagella
2. Monotrichous- flagella located at one pole
3. Amphitrichous- single flagella at each pole
4. Lopotrichous- tuft of flagella at both poles
5. Peritrichous- flagella around the cell
6. Cephalotrichous-- two or bunch of flagella are attached to one
end/pole.
b. PILI
- shorter and straighter than flagella;
- composed of subunits of a protein called pilin.
- found mainly on gram-negative organisms
- one of the important functions of a pili is to form attachment between male and
female bacteria during conjugation. A pili of this type is called a Conjugative pili/
sex pili.

c. ENDOSPORES
- A resistant asexual spore that develops inside some bacteria cells.
- Metabolically inactive bacterial cells that are highly resistant to dessication, heat,
UV and Gamma radiation and various chemicals.
- contain calcium dipicolinate, which aids in heat resistance in the core
- helpful to identify some species of bacteria,
- Examples:
- Bacillus and Clostridium Specialized DNA – allowing to survived without
nutrient
- Cortex - contains an inner membrane known as the core. The inner
membrane that surrounds this core leads to the endospore's resistance
against UV light and harsh chemicals that would normally destroy microbes
- DNA repair enzymes – repair damaged during germination.

WITHIN:
A. CYTOPLASM
- Roles:
- Protein synthesis
- energy production
- signal transduction
- transportation of metabolites and molecules from organelles across the cell
- provision of structural support to organelles of the cell
- regulation of cell signaling
- structural support for the cell itself.
- The cytoplasm of both eukaryote and prokaryote cells consists of a gelatinous liquid known
as cytosol. The cytosol is made up of a mixture of colloidal proteins which include
enzymes, carbohydrates, small protein molecules, ribosomes and ribonucleic acid (RNA).
- Eukaryotes: External to the nuclear membrane and internal to the cellular membrane.
- Prokaryotes: Internal to the cellular membrane.
B. NUCLEOID
- single long circular double stranded DNA molecule devoid of highly conserved histone
protein (prokaryotes only have this)
- The histone is present in eukaryotes, therefore, results the eukaryotic DNA into the
beaded structures (i.e. nucleosomes)
- Two types of nuclear bodies can be observed:
1. Envelope associated nucleoid - a large amount of RNA, proteins, lipids and
peptidoglycan are found,
2. Envelope free nucleoid. - contains less amount of RNA, proteins, lipids and
peptidoglycan are found,

- Host bacteria have a single, circular chromosome that is responsible for replication ,
although few species have 2 or more.

C. PLASMIDS
- A plasmid contains 5-100 genes that determine several biological functions. Under certain
circumstances, they provide special characteristics to the bacterial cell and help them in
survivability.
- They may even lose without harming the bacterial cell.
- Self replicating
- Plasmids are the circular DNA molecule but in resting stage helix twists in right hand
direction at every 400-600 base pairs and forms supercoils.
- The twisted form is called covalently closed circular- DNA.
- After cleaving the twists this form is converted into an open circular form of double stranded
DNA molecule
- *2 DNA possible for a bacteria/virus? : BDNA and plasmid
- Responsible for Antibiotic Resistance and Toxin Production
- 4 TYPES
1. RESISTANCE- antibiotic resistance
2. VIRULENCE- make good bacteria into harmful/pathogenic bacteria if it sense danger
(as a defense mechanism)
3. DEGRADATIVE- enzymes breaking chemical components
4. COL- only involves Escherichia coli

D. RIBOSOMES
- Microscopic “FACTORIES” found in all cells including bacteria.
- a complex cellular mechanism used to translate genetic code into chains of amino
acids
- Long chains of amino acids fold and function as proteins in cells.
- Difference
- Prokaryotic Ribosomes – some antibiotic inhibits its function thus killing the
bacteria.
- Eukaryotic Ribosomes – antibiotics will not inhibit its function ; it will not kill the
organism.

E. CYTOPLASMIC MEMBRANE
- The plasma membrane or cytoplasmic membrane functions to protect the watery, gel-like
interior of the bacterial cell. It completely surrounds the cell and is mostly made of fat
and protein molecules.
- Main functions of the bacterial plasma (cytoplasmic) membrane:
- Protects the inner components of the cell
 - Selective permeability (which also allows nutrients to enter and waste to exit) (By
PHOSPHOLIPID BILAYER)
- Materials move across the bacterial cytoplasmic membrane by : PASSIVE
DIFFUSION, ACTIVE DIFFUSION, CYTOLYSIS:
I. PASSIVE DIFFUSION
- the diffusion of substances across a membrane.
- spontaneous process and cellular energy is not expended.
- Molecules will move from where the substance is more concentrated to where it is less
concentrated.

II. FACILITATED DIFFUSION

- is a type of passive transport that allows substances to cross membranes with the
assistance of special transport proteins.
- Some molecules and ions such as glucose, sodium ions, and chloride ions are unable
to pass through the phospholipid bilayer (hydrophobic and hydrophilic) of cell
membranes. By the use of ion channel proteins and carrier proteins that are embedded in
the cell membrane, these substances can be transported into the cell.
● CHANNEL PROTEINS (AQUAPORINS) –allows water to diffuse at a very fast rate.

III. OSMOSIS
- is a special case of passive transport. In osmosis, water diffuses from a hypotonic (low
solute concentration) solution to a hypertonic (high solute concentration) solution.
- Generally speaking,the direction of water flow is determined by the solute concentration
and not by the nature of the solute molecules themselves.
- TYPES:
- HYPERTONIC -Fluid would flow from the area of low solute concentration (the
blood cells) to an area of high solute concentration (water solution). As a result, the
blood cells will shrink.
- ISOTONIC - Fluid would flow equally between the blood cells and the water
solution. As a result, the blood cells will remain the same size.
- HYPOTONIC - Fluid would flow from the area of low solute concentration (water
solution) to an area of high solute concentration (the blood cells). As a result, the
blood cells will swell and may even burst

IV. ACTIVE TRANSPORT

- Active transport is the process of transferring substances into, out of, and between cells,
using energy.
- Active transport is most commonly accomplished by a transport protein that undergoes a
change in shape when it binds with the cell’s “fuel,” a molecule called (ATP).
- TYPES OF ACTIVE TRANSPORT
- ANTIPORT PUMPS - Antiport pumps are a type of transmembrane co-transporter
protein. They pump one substance in one direction, while transporting another
substance in the opposite direction.
- SYMPORT PUMPS - a substance that “wants” to move from an area of high
concentration to low concentration down against its concentration gradient is used to
“carry ” another substance against its concentration gradient.
- ENDOCYTOSIS – cell uses proteins in its membrane to fold the membrane into the
shape of a pocket. This pocket forms around the contents to be taken into the cell.

V. CYTOLYSIS
- Also known as osmotic lysis,
 - occurs when a cell bursts and releases its contents into the extracellular environment due
to a great influx of water into the cell, far exceeding the capacity of the cell membrane to
contain the extra volume.
- This is a concern particularly for cells that do not have a tough cell wall to resist internal
water pressure.

- TYPES OF CYTOLYSIS
- ENDOCYTOSIS--large items, or large amounts of extracellular fluid, may be taken into a
cell by engulfing
- Receptor- mediated - a cell’s receptor may recognize a specific molecule that the cell
“wants” to take in,and form a vesicle around the area where it recognizes the
molecule.
- In other types of endocytosis, the cell relies on other cues to recognize and engulf a
particular molecule.
- EXOCYTOSIS- the cell creates a vesicle to enclose something inside the cell, for the
purpose of moving it outside of the cell, across the membrane.
- This most commonly occurs when a cell wants to “export” an important product, such
as cells that synthesize and export enzymes and hormones that are needed
throughout the body

IX. FUNCTIONS ASSOCIATED WITH BACTERIAL CYTOPLASMIC MEMBRANE &

DIVISOME

DIVISOME

-
Bacterial cells are critically dependent on their ability to divide.
-
large and highly dynamic molecular machine that carries out the process of division
-
“If it divides, it multiplies”
-
Roles
- Energy production
- Contain bases of bacterial flagella used in motility.
- Waste removal
- Formation of endospore
-
X. MORPHOLOGY
A. COCCI
- meaning "berries"; spherical, oval, or flattened on one side
● cocci in clusters- Staphylococcus sp.
● cocci in chains- Streptococcus sp.
● cocci in pairs(diplococci) w/ pointed ends- Streptococcus pneumoniae
● cocci in pairs(diplococci) w/ kidney bean shape- Neisseria sp.
● cocci in groups of four(tetrads)- peptococcus (micrococcus)
● cocci in packets of eight(cuboidal)- Sarcina sp
-
B. BACILLI
- rod shaped bacteria
● rods with squared ends- Bacillus sp.
● rods with round ends- Salmonella sp.
● club-shaped- Corynebacterium sp/
● fusiform-Fusobacterium sp.
● comma-shaped-Vibrio sp.
-
 C. SPIROCHETES
- spiral in form
● tightly-coiled - Leptospira sp. and Treponema sp.
● relaxed/loosely -coiled - Borrelia sp.
-
D. PLEOMORPHIC -
- lacking distinct shape
● Mycoplasma sp.

XI. METABOLISM
- Is used to describe all the chemical reaction that occurs in a cell. Bacteria rely on enzymes
for their biochemistry.
- Endoenzymes (which work within the cell)
- Exoenzymes (produced inside the cell and then transported to the outside where they
facilitate the preliminary digestion of high molecular weight substrates that do not pass
readily through the cell membrane)

● Lipolytic Bacteria- hydrolyzes triglycerides due to the production of extracellular

lipases. ( e.g. Staphylococcus, Micrococcus, Pseudomonas )
● Saccharolytic Bacteria – metabolizes complex CHO under aerobic and anaerobic
conditions and liberate energy. ( e.g. Bacillus, Clostridium, Aeromonas, Pseudomonas)
● Proteolytic Bacteria– they produce protease enzyme. (e.g. some in the family
Enterobacteriaceae , Clostridium, Flavobacterium)

XII. METABOLIC CHARACTERISTICS

● OXYGEN
- molecular O2 is very reactive
- it can form hydrogen peroxide (H2O2), superoxide radicals, singlet oxygen, and hydroxyl
radical (OH-)
- Enzymes possessed by bacteria to break down oxygen products:

a. Catalase- breaks down hydrogen peroxide

b. Peroxidase- breaks down hydrogen peroxide

c. Superoxide dismutase-breaks down the superoxide radical

XIII. GROWTH CHARACTERISTICS: Classification of organisms based on O2 requirement and

their characteristics

ORGANISMS O2 REQUIREMENT ENZYMES PRESENT

1. Obligate aerobe can't survive without oxygen catalase, peroxidase,

superoxide dismutase

2. Facultative anaerobe primarily aerobic, but can catalase, superoxide

grow in the absence of oxygen dismutase

3. Microaerophilic only aerobic growth; O2 superoxide dismutase

anaerobe required at low conc. ;grow
 maximally at pO2 of <0.5% to
0.3%

4. Aerotolerant anaerobe only anaerobic growth, superoxide dismutase

continues growth in the
presence of O2

5. Obligate anaerobe can't live with oxygen none

XIV. NUTRITION
A. CARBON AND ENERGY SOURCE
1. Phototrophs- use light as an energy source
2. Chemotrophs- use chemical compounds as an energy source
3. Autotrophs- (aka litotrophs) use inorganic sources such as ammonium and sulfide as an
energy source
4. Heterotrophs- (aka organotrophs) use organic carbon (lipids, proteins and carbohydrates)
as an energy source
5. Chemoheterotrophs- use chemical and organic compounds such as glucose for energy -
include all medically important bacteria
6. Capnophilic organisms- prefer an environment with increased CO2 (3%-10%)
7. Salt loving organisms- Staphylococcus aureus survives high salt concentration 7.5% -
Vibrio sp.

II. TEMPERATURE REQUIREMENT

A. Psychrophilic(cold loving) - grow below 10°C (found in refrigerators or blood bags or ICU
and may cause pneumonia; -ex: Listeria monocytogenes, Yersinia enterolitica
B. Mesophilic- grow at 20-40°C; best at 30-40°C
C. Thermophilic( heat loving)- grow at 50-55°C
D. Hyperthermophilic- optimum growth at 80°C or more; most of the organisms live in the
hot springs associated. with volcanic activity and sulfur usually important for their
metabolic processes.

XV. STAINING CHARACTERISTICS

- the morphology and staining rxn of the organism are significant clues to its identity
A.GRAM STAINING METHOD
- Hans Christian Gram developed the Gram staining method in 1884.
- Gram staining procedure uses four chemicals; crystal violet, iodine, alcohol,
and safranin, to stain bacteria.
- This differential staining technique separates most bacteria into two groups based
on cell wall composition”
1. Gram-positive bacteria stains purple or blue
● GRAM POSITIVE (+) ORGANISMS
- Principle : Involves the ability of the bacterial cell wall to retain the crystal violet dye
during solvent treatment.
- Gram Positive microorganisms have THICK LAYER of PEPTIDOGLYCAN ( TECHOIC and
LIPOTEICHOIC ACID complexes which are INSOLUBLE to Alcohol Decolorizer, the CV
Complex remains intact giving blue-violet color.
- (e.g. Staphylococci, Streptococci, Pneumococci, C.diphtheriae, B. anthracis)

2. Gram-negative bacteria stains red/pink

● GRAM NEGATIVE (-) ORGANISMS
- Principle : The cell wall is composed of thin layer of a particular substance
PEPTIDOGLYCAN covered by an outer membrane of LIPOPROTEIN and
LIPOPOLYSACCHARIDES containing ENDOTOXIN which is INSOLUBLE to Acid
decolorizer, it losses the CV Complex giving pink – red in color.
- (e.g. E.coli, Legionella, Neisseria spp.)
 ● REAGENTS:
ØCrystal Violet – primary stain.
ØGrams Iodine - mordant ( strengthens the affinity of the bacteria with the CV
ØAcetone Alcohol – Decolorizer
ØSafranin – Counter stain

● STEPS:
1. Bacterial Smear Preparation
a. Take a clean grease free slide.
b. Transfer a loop of the sample (for example, sputum, CSF, or pus) to the
microscope slide. If performing a Gram stain from a bacterial colony, first put
a drop or a few loopful of water and emulsify the bacterial colony in the water
drop.
c. Spread the sample to an even-thin film.
d. Air dry the sample, and once the sample gets air dried,
e. heat fix the smear by passing it through a flame three times.

2. Staining Procedures
a. Flood the heat-fixed smear with CV for 1min.
b. Rinse with tap running water.
c. Flood with the mordant for 1min
d. Wash slides indirectly with tap water until the excess comes off.
e. Flood it with decolorizer .
f. Rinse gently with water
g. Flood slide with counterstain, safranin.
h. Rinse slide gently and Air dry.

B. ACID FAST STAIN

- a differential stain used to identify acid-fast organisms such as members of the genus
Mycobacterium and Nocardia.
- Principle :The primary stain used in acid-fast staining, carbolfuchsin, is lipid-soluble and
contains phenol, which helps the stain penetrate the cell wall. This is
further assisted by the addition of heat. The smear is then rinsed with a very strong
decolorizer, which strips the stain from all non-acid-fast cells but does not permeate the cell
wall of acid-fast organisms. The decolorized non-acid-fast cells then take up the
counterstain. ( e.g.Mycobacterium spp)

● REAGENTS:
ØCarbol Fuchsin – primary stain
ØPhenol (Carbol fuchsin mixture) – mordant
ØAcid Alcohol ( 90-95 % with HCl 3-5 %)- decolorizer
ØMethylene Blue or Brilliant Cresyl Green - counterstain

● STEPS:
1. .Air dry and heat fix a thin film of microorganisms. Allow the slide to cool.
2. Flood the slide with Carbolfuchsin. Steam the slide with a Bunsen burner over the sink.Let
the slide set for 5 minutes. Rinse with water.
3. Flood slide with Acid Alcohol for 30 seconds. Rinse with water.
4. Counterstain by flooding the slide with Methylene Blue for 30 seconds. Rinse with water.
 5. Air dry
6. View organisms using the oil immersion objective of your microscope.

XVI. COLONY CHARACTERESTCS

COLONY :
- visible mass of microbial cell.
- A single bacterial colony indicates a group of bacterial cells or a bacterial mass.
- In a colony, all the bacterial cells originate from a single mother cell and look identical
to each other.

COLONIAL MORPHOLOGY : distinct characteristics of the microbial mass formed on the nutrient
base. The distinctive features of a microbial colony include shape, elevation, color, margin etc

● SIZE: Punctiform, Small, Moderate, Large

● COLOR: Orange, Red-pink, Black, Brown, White
● OPACITY: Transparent, Translucent, Opaque, Iridescent
● TEXTURE: Slimy, moist, Matte, brittle, Shiny, viscous, Dry, mucoid
● SHAPE: Circular, Punctiform, Irregular, Filamentous, Rhizoid, Spindle
● ELEVATION: Raised, Convex, Flat, Umbonate, Crateriform, Pulvinate
● MARGIN: Entire, Undulate, Filiform, Curled, Lobate, Erose

Other Notes:
● CHO- carbohydrates
● CHON- protein
● Virulence factor- severity of how causative agents could harm you
● Salmonella typhi associate with Mary Mallon “Typhoid Mary”; tanan kilutuan namatay
● Virus- does not have cytoplasm and only has DNA and RNA which can be single or double
stranded. Does not have ribosomes