Micro Lab

Microbiology
Laboratory ONLINE Class
MPH@2020
Microbiology
Laboratory
Safety Rules
Lesson 1
MPH@2020
Microbiology Laboratory
Safety Rules
●
Never work alone in the laboratory without
permission and prior knowledge of the
instructor.
●
Do not engage in rowdy, playful, or
unprofessional activities in the laboratory. This
includes not being disrespectful of your
instructor or classmates.
●
Work surfaces must be disinfected at the
beginning and at the end of every laboratory
period.
https://www.actx.edu/biology/microbiology-laboratory-safety-rules#: MPH@2020
Safety Rules
4. All students must wash their hands at the
beginning of lab and at the end of every
laboratory period.
5. No eating or drinking is permitted in the
laboratory. No open food or beverage can be
taken into the laboratory.
6. Lab benches are to be kept free of extraneous
items while conducting experiments. This
includes personal items such as backpacks, cell
phones, and unnecessary books.
Safety Rules
7. Students must wear closed-toe shoes that
cover the top of the foot, and appropriate
clothing, at all times in the laboratory.
8. Students must wear examination gloves
when handling microorganisms. Wear lab
aprons or lab coats as advised by your
instructor. Wear safety glasses when
handling bacterial broth cultures, doing Gram
stains, and as otherwise advised by your
instructor.
Safety Rules
9. Keep hands away from your face, eyes, and
mouth when working with chemicals or
microorganisms. This includes not applying
cosmetics, not adjusting contact lenses, and
not biting your finger nails.
10. If any chemicals or other agents splash into
your eyes, immediately go to the nearest sink
and flush your eyes with water.
Safety Rules
11. Take special precaution when working with open
flames. Loose hair, clothing, dangling jewelry,
and nearby paper must be secured. Do not
leave a heat source (hot plate or Bunsen burner)
unattended. Keep containers of alcohol,
acetone, or other flammable liquids at a safe
distance from flames.
12. Report ANY and ALL accidents, spills,
BREAKAGES, or injuries to the instructor, no
matter how trivial they appear.
Safety Rules
13.Any pregnant or immunocompromised student must notify
the instructor of the course. A pregnant student is required
to wear safety glasses and 2 sets of examination gloves
when handling any bacterial broths or cultures.
14.Do not remove cultures, reagents, or other materials from
the laboratory unless specific permission from the instructor
has been granted.
Safety Rules
15.Do not use any lab equipment without instruction and
authorization from the instructor. Report any damaged or
broken equipment to your instructor immediately.
16.Students must assume that all of the organisms that they
work with in this laboratory are potential pathogens
(disease-producing microbes). However, our laboratory
does not require the use of any highly infectious human
pathogens.
Proper Waste
Disposal
MPH@2020
Proper Waste Disposal
1. Any student's laboratory manual or

personal paper must be secure not to
contaminate with microorganisms.
2. Do not dispose of cultured plates and agar
plates in the sink or wastebasket. Before
disposal, culture plates must be autoclave.
3. Disposed of in a yellow-colored waste bag
all contaminated gloves and other materials
or biohazard waste container.
4. Wrap with box broken contaminated glass and label before

autoclaving. Place broken uncontaminated glass to the
designated receptacle for broken sharps and use a dust-pan
when cleaning up broken glassware.
5. Disposed biohazardous wastes in a biohazard waste
container.
6. Students must not leave the laboratory and must not touch
any equipment such as microscopes, any personal items
such as cell phones, or any doorknobs while wearing
examination gloves.
7. Disposed uncontaminated gloves in the regular trash and
disposed of contaminated gloves in a biohazard waste
container.
6. Students must not leave the laboratory and must not touch
any equipment such as microscopes, any personal items
such as cell phones, or any doorknobs while wearing
examination gloves.
7. Disposed uncontaminated gloves in the regular trash and
disposed of contaminated gloves in a biohazard waste
container.
Activity 1
Microbiology Laboratory Safety Rules
1.Write down five more laboratory safety rules. Explain

each.
2.Add five ways in disposing of microbial contaminated
items and explain each.
3.Attach illustration to each laboratory rules and proper
waste disposal.
MPH@2020
Laboratory
Equipment
Use in
Microbiology
Lesson 2
MPH@2020
Laboratory Equipment Use in
Microbiology
MPH@2020
Microbiology
MPH@2020
Microbiology
Inoculating loop Inoculating needle Petri Dish
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Activity 2:
Laboratory Equipment Use in Microbiology
1.Add all additional laboratory equipment and

apparatus needed in the microbiology laboratory.
2.Provide pictures of each laboratory equipment and
apparatus.
3.Give the use of each equipment and apparatus.
4.Give the proper aseptic technique before and after
using these equipment and apparatus.
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Control of
Microorganisms
Lesson 3
MPH@2020
Control of Microbial
Growth
Control of microbial growth in the
pharmaceutical preparation is crucial.
Microbial growth affects the overall
condition of the product. Product
deterioration, loss of potency, and initiate
infection to the user are some of the
effects of microbial contamination.
Prevention of contamination is being done,
such as aseptic technique and disinfection
(D'Ugo,2016).
MPH@2020
Growth
The control of microbial growth is significantly
important not only in the pharmaceutical industry
but as well as in food and agriculture
(Alnaimat,2012).
Two basic ways:
1.By killing microorganisms or
2.By inhibiting the growth of microorganisms
MPH@2020
Growth
Control of growth usually involves
the use of physical or chemical
agents which either kill or prevent
the growth of microorganisms
(Alnaimat,2012).
This could be:
1.Bactericidal
2.Bacteriostatic
MPH@2020
Growth
Agents which kill cells are called cidal agents;
agents which inhibit the growth of cells (without
killing them) are referred to as static agents
(Alnaimat,2012).
Sterilization is the complete destruction or
elimination of all viable organisms (in or on an
object being sterilized). Sterilization procedures
involve the use of heat, radiation or chemicals, or
physical removal of cells.
MPH@2020
Growth
Methods of Sterilization
(Alnaimat,2012).
Heat: most important and widely used..
1.Incineration: burns organisms and
physically destroys them. Used for
needles , inoculating wires, glassware,
etc. and objects not destroyed in the
incineration process.
MPH@2020
Growth
(Alnaimat,2012).
2.Boiling: 100°C for 30 minutes. Kills
everything except some
endospores. To kill endospores,
sterilize the solution, very long or
intermittent boiling is required.
MPH@2020
Growth
(Alnaimat,2012).
3.Dry heat (hot air oven): 160°C/2hours
or 170°C/1hour. Used for glassware,
metal, and objects that won't melt.
MPH@2020
Growth
Methods of Sterilization (Alnaimat,2012).
4.Pasteurization : is a process of heating a food, which is
usually a liquid, to a specific temperature for a predefined
length of time and then immediately cooling it after it is
removed from the heat. This process slows spoilage due to
microbial growth in the food. Unlike sterilization, pasteurization
is not intended to kill all microorganisms in the food. Instead, it
aims to reduce the number of viable pathogens so they are
unlikely to cause disease.
MPH@2020
Growth
5.Autoclaving (steam under pressure
or pressure cooker): 121°C for 15
minutes (15 Ib/sq pressure). Good for
sterilizing almost anything, but heat-labile
substances will be denatured (degrade)
or destroyed.
MPH@2020
Growth
Irradiation: usually destroys or distorts
nucleic acids. Ultraviolet light is usually
used (commonly used to sterilize the
surfaces of objects), although x-rays and
microwaves are possibly useful.
MPH@2020
Growth
Filtration: involves the physical removal
(exclusion) of all cells in a liquid or gas,
especially important to sterilize solutions
which would be denatured by heat (e.g.
antibiotics, injectable drugs, amino acids,
vitamins, etc.)
Denaturation is the alteration of a protein shape through some form of external stress (for example, by
applying heat, acid or alkali), in such a way that it will no longer be able to carry out its cellular function.
MPH@2020
Growth
Chemical and gas:
Alcohol 70% : Antiseptics and disinfectants
Formalin:( Formaldehyde) is an organic compound with the formula CH2O,
used as disinfectants.
Ethylene oxide gas : This highly reactive gas (C2H4O) is flammable, toxic,
and a strong mucosal irritant, used for contaminated medical tools.
Halogens: Chlorine, iodine, and derivatives of these halogens are suitable
for use as disinfectants. Chlorine and iodine show a generalized
microbicidal effect and also kill spores
MPH@2020
The Effectiveness of Hand
washing
In the microbiology laboratory, handwashing
is an effective way to protect the microbial
cultures, the staff, and the community.
A laboratory session should start with
washing hands with soap or detergent to
remove transient microbes that might
contaminate culture.
Likewise, the last thing to do before leaving the area is to wash
thoroughly and disinfect the hands to remove pathogens.
Laboratory Manual in Microbiology and Parasitology by Bognot; 7th ed,2015 MPH@2020

washing
Proper handwashing by clinical personnel is
the most effective method of controlling
infections, especially nosocomial ones.
The structure of and layer of oil on the skin

prevent the removal of microorganisms.
Simple handwashing using soap helps

remove oil while scrubbing with a brush for 7
– 8 minutes eliminates both transient and https://www.youtube.com/watch?
v=3PmVJQUCm4E
resident microbes.
Laboratory Manual in Microbiology and Parasitology by Bognot; 7th ed,2015 MPH@2020
washing
https://www.youtube.com/watch?v=3PmVJQUCm4E MPH@2020
Activity 3:
Proper Hand Washing
Do the proper hand washing procedure at home and send a

picture while doing the procedure.
Answer the following:
1. Is there a proof that hand washing by health personnel

can help reduce the patient's risk of contracting nosocomial
infection? Explain your answer.
2. Enumerate the steps in proper hand washing.
MPH@2020
The
Microscope
Lesson 4
MPH@2020
The Microscope
The early pioneers of microscopy opened a window into the

invisible world of microorganisms. But microscopy continued
to advance in the centuries that followed.
In 1830, Joseph Jackson Lister created an essentially
modern light microscope.
The 20th century saw the development of microscopes that
leveraged nonvisible light, such as f luorescence microscopy,
which uses an ultraviolet light source, and electron
microscopy, which uses short- wavelength electron beams.
MPH@2020
Parts of
Microscope
MPH@2020
Computing total
Magnification
The ocular lenses typically magnify images 10 times (10 ). At the
other end of the body tube are a set of objective lenses on a
rotating nosepiece. The magnif ication of these objective lenses
typically ranges from 4 to 100 , with the magnif ication for each
lens designated on the metal casing of the lens. The ocular and
objective lenses work together to create a magnif ied image. The
total magnif ication is the product of the ocular magnif ication times
the objective magnif ication:
Total magnif ication = ocular magnif ication × objective magnif ication
Ex. TM = (10x) x (40x); TM = 400x
Molly Smith, Lab Manual: Microbiology for Allied Health Students, 2018 MPH@2020
How to use a microscope?
1.Turn the revolving nose piece so that the lowest power objective lens
(eg. 4x) is clicked into position.
2.Place the microscope slide on the stage and fasten it with the
stage clips.
3.Look at the objective lens and the stage from the side and turn
the focus knob so the stage moves upward. Move it up as far as
it will go without letting the objective touch the coverslip.
4.Look through the eyepiece and move the focus knob until the
image comes into focus.
https://www2.mrc-lmb.cam.ac.uk/microscopes4schools/microscopes2.php MPH@2020
5.Adjust the condenser and light intensity for the greatest amount of
light.
6.Move the microscope slide around until the sample is in the
centre of the f ield of view (what you see).
7.Use the f ine focus knob to place the sample into focus and
readjust the condenser and light intensity for the clearest image
(with low power objectives you might need to reduce the light
intensity or shut the condenser).
8.When you have a clear image of your sample with the lowest power
objective, you can change to the next objective lenses. You might
need to readjust the sample into focus and/or readjust the condenser
and light intensity. If you cannot focus on your specimen, repeat
steps 3 through 5 with the higher power objective lens in place. Do
not let the objective lens touch the slide!
9.When f inished, lower the stage, click the low power lens into
position and remove the slide.
Youtube: https://www.youtube.com/watch?v=SUo2fHZaZCU MPH@2020

Activity 4
1.Enumerate the parts of the microscope and give its
functions
2.When is the oil immersion objectives used? What kind of oil
is used?
3.When is the concave surface of the mirror used?
4.Name other types of microscope and discuss the principles
involved in their application?
5.What is the differences between brightfield and darkfield
microscopy?
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Morphology and
living state of
microorganisms
Lesson 5
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Morphology of Bacteria
Cocci
S.Aureus
Microscopic view without stain
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Bacilli
E.Coli
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Vibrio
Vibrio cholerae
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Spirochete
Treponema palladium
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Microorganisms in Living
State
Bacteria move by means of flagella
whereas protozoans use either the flagella,
cilia, or pseudopods for movement.
True motility should not be mistaken for
Brownian movement which is the random
movement produced by currents in liquids
causing organisms to vibrate and move.
Motile organisms can be observed in their Pseudomonas using a single polar
natural living state using wet mount flagellum for movement
method or hanging drop preparation. http://www.youtube.com/embed/EWj2T

GsTQEI
MPH@2020
Microorganisms in Living
State
The wet mount method involves placing a
drop of specimen on a glass slide and
covering it with a slip.
In hanging drop method, a cover slip with a
drop of the specimen is inverted over a
slide with concavity at the center (hanging
drop slide).
Both preparation are examined under the
microscope.
MPH@2020
Microorganisms in Living State
https://www.facebook.com/100001512
735584/videos/3701216496605405/
MPH@2020
Activity 5
Morphology and living state of
microorganisms
1.Give five examples of microorganisms base on shape.
(Cocci, Bacilli, vibrio, spirochete)
2. Aside from observing motility, what are the other uses of
the wet mount and hanging drop method?
3. Prokaryotic organisms like bacteria use the flagella for
movement. What is the basic structure of a flagellum?
4. Aside from flagella, the other structure for motility are
pseudopods. What are pseudopods? Name some
organisms using these structure for movement.
MPH@2020
Staining
Techniques in
Microbiology
Lesson 6
MPH@2020
Staining Technique in
Microbiology
Studying bacteria and other microorganisms in their natural
state can be difficult. Aside from being extremely small,
bacteria and microorganisms are also color less and
transparent when examined under the microscope.
To visualize them, stains or dyes are used to impart color and
provide contrast to their surroundings.
Stain used in microbiology are either basic, acidic, or neutral.
They are salt composed of a positive and a negative ion.
Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

Staining Technique in Microbiology
The chromatophore is the colored ion that impart color.

In general, dyes are classified as acidic if the chromatophore
has a negative charge. Acid dyes react with the positively
charge components of the cell like the cytoplasm. Because of
this, acidic dyes are referred to as cytoplasmic dyes.
Example:
1.Nigrosin
2.India ink

On the other hand, Basic dyes, also called nuclear stains, have
positive charged chromophores that stain those cell parts that
are negatively charged.
Neutral stains, on the other hand, consist of a mixture
Basic dyes includes: of acidic and basic dyes.
1.Crystal violet
2.Methylene blue
3.Safranin
4.Malachite green

There are three staining techniques commonly used.

This are the:
1.Simple ( commonly used stain = crystal violet or any basic
dye)
2.Differential (step by step: crystal violet, gram's iodine and
safranin)
3.Special or structural staining ( step by step: malachite
green and safranin)

https://www.youtube.com/watch?v=n5fXIpJUgD4
Simple staining: Prior to the application of stain, a thin film

of organisms is prepared, dried and fixed
a single basic dye is used to onto a clean slide. This is called a smear
color and highlight the entire
organism.
Bacteria smear
https://www.youtube.com/watch?v=on5-5oQNNqo
Differential staining:
Distinguish bacteria according to their reaction to particular
stains. An example of very useful differential staining technique
is the gram stain procedure that classified bacteria as either
Gram-positive or Gram negative.
https://www.youtube.com/watch?v=AZS2wb7pMo4

Special or structural staining:

Use to color a specific part or
structure of microorganism, like
the endospore, flagellum or
capsule
https://www.youtube.com/watch?v=JVn6ZRFtARs
Activity 6:
Make a table of all bacteria belong to gram-negative and

gram-positive. (Observe proper writing of scientific names)
Example table:
Gram-positive bacteria Gram negative bacteria
Stain color: blue Stain color: pink
1. Staphylococcus aureus 1. Escherichia coli
2. 2.

Microbiological
Culture Media
Preparation
Lesson 7
MPH@2020
Microbiological Culture Media
preparation
Microbes, including bacteria and fungi, need nutrients like
carbon, nitrogen and other growth factors in order to survive.
In the laboratory, the nutrient preparations for culturing
microorganisms are called media.
This media maybe in:
1.Liquid
2.Semi-solid
3.Solid form

preparation
The major difference among them is the presence of
a solidifying agent (agar).
Liquid Culture Media (broth)
- Use to grow microbes at a high cell density
Semi-solid Culture Media
- is jelly-like due to the lower concentration of
solidifiers
- it is useful in determining bacterial motility as
well as isolating microbes
preparation
Solid Culture Media (agar plate)
- is used for surface growth to observe colony
formation and for pure culture isolation (nutrient
agar, blood agar)
A broth medium, when supplemented with agar,
becomes a solid medium that can be placed in a
petri dish.

preparation
Agar slant
When a liquid medium containing agar is poured
into a test tube and allowed to harden in a slant
position.
Agar deep or agar butt
If the medium is allowed to harden in an upright
position. Similar to a slant but creates a deep zone
for aerobic and anaerobic microorganism.

preparation
All culture media must be sterilized and be free from other
forms of life including endospore. There are three basic ways
of sterilizing media and other supplies, namely:
1.Autoclaving
2.Dry heat sterilization
3.UV radiation
The most common approach is autoclaving, which involves
sterilizing items and media through moist heat or steam at
121ºC, in a pressure of 15 psi (pounds per square inch) for
15 minutes or longer, depending on the item.
preparation
Preparation of
Agar Plate
https://www.youtu
be.com/watch?
v=cneascR3OEc
Activity 7
Microbiological Culture Media preparation
Give the step by step procedure in the preparation of the
following culture media:
1. Nutrient broth (NB)
2. Nutrient agar (NA)

Inoculation
Method: Aseptic
Culture
Techniques
Lesson 8
MPH@2020
Inoculation method: Aseptic Culture
Techniques
In the laboratory, microbes must be cultured in
order to facilitate identification and to examine
their growth and metabolic characteristics.
Microbes are inoculated or introduced into
various forms of culture media to keep them
alive for study.
Inoculation must be done through aseptic technique to prevent
contamination with unwanted microbes or contaminants.
This technique usually involves:

Techniques
This technique usually involves:
1. Disinfecting the working areas
2. Minimizing contacts with contaminates
3. Using flame to eliminate microbes which
might enter vessels such as test tubes and
petri dish when they are opened.

Techniques
There are three inoculation methods used for
isolation of bacteria:
1. Streak plate method
2. Spread plate method
3. Pour plate method
Spread plate and Pour plate method are quantitative

techniques that allows determination of microorganisms
population in a sample.

Techniques
Streak plate method is the most common
isolation technique that uses sterile, heat
resistant, and none-corroding nichrome wire
attached to an insulated handle called
inoculating loop or inoculating needle in
transferring bacteria in different culture media.
For special purposes, cultures may also be transferred with

sterile cotton swabs, pipettes, glass rods, or syringes,
depending on the medium and technique.

Techniques
https://www.y
outube.com/w
atch?
v=bRadiLXkq
oU

Activity 8
Inoculation method: Aseptic Culture Techniques
Answer the following questions:

1. What is the importance of sterilizing the inoculating loop
and needle before and after the inoculation process?
2. What is the purpose of flame-sterilizing the mouth of the
test tube and the side of the petri dish during inoculation?
3. How can one be sure that the culture media are sterile or
free from contamination?
4. What are the precautionary measures to be observed
before, during and after applying the aseptic culture technique?
Bacterial
Culture
Characteristics
in Different
Microbiological
Media
Lesson 9
MPH@2020
Bacterial Culture Characteristics in
Different Microbiological Media
The cultural characteristics of organisms like bacteria pertain
to their macroscopic appearance on different kinds of culture
media.
Microbes are cultured or grown into culture media to increase
their number during the incubation period.
After a suitable period, some of the dispersed cells develop
into colonies.
A colony is a population of cells arising from a single bacterial
cell in a solid medium. It is visible to the naked eye.

Bacterial growth in different culture media is characterized by
its appearance.
Colonial characteristics and the amount of growth after
incubation are essential information in the identification of
bacteria.
The morphology of colonies can be described by size, shape,
color, edge, or margin.
These features are observed by looking at the top of the
colony itself

Colony elevation is apparent when viewed from the sides at
the eye level.
The growth of bacteria in broth is described based on the
turbidity of the broth after incubation.

http://www.medical-labs.net/bacterial-colony-morphology-2-887/ MPH@2020
Some examples of SHAPES:
Circular Irregular Punctiform
http://www.uwyo.edu/molb2021/virtual-edge/lab01/colony_morphology.html MPH@2020
Some examples of ELEVATION:
Convex Umbonate https://www.morton-

pub.com/sites/default/files/promos/customl
Elevation of bacterial colonies ab_cover_wpages_micro.pdf
may be thin or thick.

http://www.uwyo.edu/molb2021/virtual-edge/lab01/colony_morphology.html MPH@2020
Margin or edge and forms:
The periphery of bacterial colonies may have different patterns
depending on the species:
Some examples of MARGIN:

Size:
Only isolated or well spread colonies should be measured. The
time at which the measurement is made must be noted since
young colonies are smaller than older ones.
(a) Pinpoint – extremely small colony, measuring
only a fraction of milliliters in diameter
or less than 1 mm.
(b)Large – colonies measuring 5 to 10 mm in
diameter.

Surface texture
● Smooth – shiny and glistering
● Rough – dull, granular, matte (flat w/o shine)
● Mucoid – slimy, gummy
● Wrinkled – crumpling, folding

Consistency:
This growth feature can be determined by touching the colony
USING an INOCULATING NEEDLE.
● Butyrous – the colony has a butter-like consistency
● Viscous or stringly – a portion of the colony may come off the
agar surface
● Rubbery – the whole colony comes off the agar surface.
● Dry, brittle, or powdery – the colony breaks when touch by
needle.

Optical feature:
● Opaque – impenetrable to light (not transparent)
● Transparent – a clear image is seen and as if there is no
intervening material.
● Translucent – permits the passage of light.

Activity 9:
Bacterial Culture Characteristics in Different
Microbiological Media
Group Activity:
Submit a slide presentation of
each example bacterial colony
base on forms, elevation, and
margin on the table.

Chemical
Control of
Microorganisms
Lesson 10
MPH@2020
Chemical Control of Microorganisms
The growth of ( potentially harmful)

microorganisms can be controlled using
various chemicals such as bleach,
ammonia, chlorine, iodine and alcohol.
Chemical that control the growth of
microbes are divided into two main
groups, namely antiseptic and
disinfectants.

Antiseptic and disinfectants are non-

selective in their action, hence inhibiting
or killing all types of bacteria unlike other
chemical agent such as antibiotics that
target specific bacterial cells.
Antiseptic: used on living tissues such
as skin to reduce the population of
microbes present.

Disinfectants are potentially harmful,

are used on inanimate objects (floor,
countertops).

Relative effectiveness of various chemical agents against

bacterial cell may be evaluated using filter paper disk method.
This test determine which chemical agent is effective in certain
situation.

Activity 10:
1. Give examples of Antiseptic.

2. Give examples of Disinfectants.
3. Enumerate the characteristics of a good antiseptic.
4. Enumerate the characteristics of good disinfectants
according to the CDC.

The Kirby-Bauer
Disk Diffusion
Method
Lesson 11
MPH@2020
The Kirby-Bauer Disk Diffusion
Method
The chemotherapeutic agents used to treat
diseases are collectively referred as
antimicrobial agents. An antimicrobial agents
is any chemical drug used to treat infection,
either by inhibiting or killing pathogens in vivo.
These antimicrobial agents include antibiotics,

substances produced by microorganisms like
bacteria or fungi effective in eliminating the
growth of other microbes.
Method
Although originally produced by
microbes, many antibiotics are now
synthesized or manufactured in
pharmaceutical laboratories due to
the increasing demands of clinical
laboratories to determine antibiotic
susceptibility or resistance of a wider
spectrum of pathogens.

Method
One of the most widely used methods to
determine the susceptibility of the
microorganisms to antimicrobial agents is the
Kirby-Bauer disk diffusion method.
The principle of this method is dependent on the
inhibition of multiplication of microbes on the
surface of a solid medium by an antimicrobial
agent which diffuses into the medium from a
filter paper.

Method
Thus, for a microbe which is truly sensitive
(susceptible) to an antimicrobial agent, a zone
of inhibition around the disc impregnated with
the agent will be observed .
Beyond this zone, an unaffected area of normal
growth (lawn) of the microbes is seen. Accurate
measurement of zone diameter in nearest
millimeter is necessary to interpret this test
property.

Method
Table of
zone of inhibition
https://www.researchgate.net/figure/Zone-diameter-interpretive-standards-chart-for-the-determination-of-antibiotic-sen_tbl1_261636878 MPH@2020
Activity 11:
The Kirby-Bauer Disk Diffusion Method
1.Differentiate an antimicrobial agent from an antibiotic.

2.Why do some microorganisms develop resistance to certain
antibiotic agents?
3.Why are there non-responsive patients to antibiotics when in
fact these antibiotics are originally effective based on clinical
sensitivity testing?

Oligodynamic
Action of Metal
Against
Bacteria
Lesson 12
MPH@2020
Oligodynamic Action of Metal
Against Bacteria
Oligodynamic action is the ability of some
metals, notably silver, zinc, and copper to kill or
inhibit the growth of microorganisms such as
bacteria when used in very dilute solution.
It means that selected metals and metal
compounds in small quantities of water have
the ability to change and finally eliminate
bacteria. Also, these metallic elements have
been observed to inactivate enzymes and inhibit
bacterial growth.
Oligodynamic Action of Metal
Against Bacteria
https://pubmed.ncbi.nlm.nih.gov/393905
7/#:~:text=subtilis%20and
%20Legionellaceae%20exhibited
%20the,rods%20was%20the%20most
%20resistant.
MPH@2020
Hospital-
Acquired
Disease:
Nosocomial
Infection
Lesson 13
MPH@2020
Hospital-Acquired Disease:
Nosocomial Infection
Hospital-acquired infections or
nosocomial infections are caused
by various bacteria, viruses, or
even fungi when a patient is
undergoing medical care.
Persons exposed to the physical
environment of a hospital like
medical and support staff, and
even people visiting the hospital
can acquire nosocomial infections.
Infections are a major problem in
most healthcare setting today.
Several studies revealed that they
can be acquired by prolonged
hospital stay or confinement that
last approximately one to two
weeks.
Manifestation may become clinically
apparent either during the hospital
stay or after discharge.
Pathogens that can cause
nosocomial infection often develop
resistance to standard antimicrobial
making therapy and control more
difficult and costly.

Micro Lab

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Microbiology

Laboratory ONLINE Class

1. Any student's laboratory manual or

4. Wrap with box broken contaminated glass and label before

1.Write down five more laboratory safety rules. Explain

Inoculating loop Inoculating needle Petri Dish

1.Add all additional laboratory equipment and

Laboratory Manual in Microbiology and Parasitology by Bognot; 7th ed,2015 MPH@2020

The structure of and layer of oil on the skin

Simple handwashing using soap helps

Do the proper hand washing procedure at home and send a

Answer the following:

1. Is there a proof that hand washing by health personnel

2. Enumerate the steps in proper hand washing.

The early pioneers of microscopy opened a window into the

Youtube: https://www.youtube.com/watch?v=SUo2fHZaZCU MPH@2020

Microscopic view without stain

Microscopic view without stain

Microscopic view without stain

Microscopic view without stain

method or hanging drop preparation. http://www.youtube.com/embed/EWj2T

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

The chromatophore is the colored ion that impart color.

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

There are three staining techniques commonly used.

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

Simple staining: Prior to the application of stain, a thin film

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

Special or structural staining:

Make a table of all bacteria belong to gram-negative and

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

Spread plate and Pour plate method are quantitative

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

For special purposes, cultures may also be transferred with

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

Answer the following questions:

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

Circular Irregular Punctiform

Convex Umbonate https://www.morton-

may be thin or thick.

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

● Viscous or stringly – a portion of the colony may come off the

● Dry, brittle, or powdery – the colony breaks when touch by

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

The growth of ( potentially harmful)

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020

Antiseptic and disinfectants are non-

Bognot, Laboratory Manual in Microbiology and Parasitology 2 nd ed;2015 MPH@2020