MB Module 1

Republic of the Philippines
Laguna State Polytechnic University

Province of Laguna
LSPU Self-Paced Learning Module (SLM)

ISO 9001:2015 Certified
Level I Institutionally Accredited

Learning Outcomes
Student Learning Strategies
Online Activities A. Online Discussion via Zoom

(Synchronous/ You will be required to attend in a 6-Hour class discussion on the
Asynchronous) scheduled topic. To have access to the Online Discussion, refer to this
link: ____________________.
The online discussion will happen on October 12 or 13 2020, 10 am-12

pm
(For further instructions, refer to your Google Classroom and see the
schedule of activities for this module)
B. Learning Guide Questions:

1. What is microbiology?
2. What are microbes?
3. How can be these microbes be beneficial to humans?
Note: The insight that you will post on online discussion forum using Learning
Management System (LMS) will receive additional scores in class participation.
Offline Activities Lecture Guide

(e-Learning/Self- History and Scope of Microbiology
Paced)
What is Microbiology?
© Image ID: 63709654
 study of organisms and agents too small to be seen by the naked eye
(<1mm)
 study of their distribution in nature, their relationship to each other and
to other living organisms, their effects on human being and on other
animals and plants, their abilities to make physical and chemical
changes in our environment, and their reactions to physical and
chemical agents
 Microbiology is about microbial cells and how they work, especially the
LSPU SELF-PACED LEARNING MODULE: TECHNOLOGY FOR TEACHING AND LEARNING

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bacteria, a very large group of very small cells that, collectively, have
enormous basic and practical importance.
 Microbiology is about diversity and evolution of microbial cells; about
how different kinds of microorganisms arose and why.
Level I It is also
Institutionally about what microorganisms do in the world at large, in soils
Accredited
and waters, in the human body, and in animals and plants.

 One way or another, microorganisms affect and support all other forms
of life, and thus microbiology can be considered the most fundamental
of the biological sciences.
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The Science of Microbiology

 understanding the living world of microscopic organisms
 applying our understanding of microbial life processes for the benefit of
humankind and planet Earth
Microorganisms are
 single-celled microscopic organisms like
Bacteria
Fungi
Algae
Protozoa
Viruses (acellular)
Microorganisms:
 First living organisms on planet
 live everywhere life is possible
more numerous than any other kind of organisms
 global ecosystem depends on their activities
 influence human society in many ways
1. First living organisms on planet

Microorganisms were the first entities on Earth with the
properties of living systems.

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The cyanobacteria had given importance in biological evolution

because of oxygen – a waste product of their metabolism –
prepared the planet for more complex life forms.
2. Live everywhere life is possible
Microbial life is all around us.
Examination of natural materials such as soil or water invariably
reveals microbial cells.
Unusual habitats such as boiling hot springs and glacial ice are
also teeming with microorganisms.
3. More numerous than any other kind of organisms
The diversity and abundance of microorganisms in microbial
communities is controlled by the resources (foods) and
conditions (temperature, pH, oxygen content, and so on) that
prevail in their habitat.
The interaction among microbial populations could be beneficial,
neutral or harmful.
For example, the metabolic waste products of one group of
organisms can be nutrients or even poisons to other groups of
organisms.
Habitats differ markedly in their characteristics, and a habitat
that is favorable for the growth of one organism may be harmful
for another.
4. Global ecosystem depends on their activities
An ecosystem is greatly influenced and, in some cases, even
controlled by microbial activities.
Microorganisms carrying out metabolic processes remove
nutrients from the ecosystem and use them to build new cells.
At the same time, they excrete waste products back into the
environment. Thus, microbial ecosystems expand and contract,
depending on the resources and conditions available
Over time, the metabolic activities of microorganisms gradually
change their ecosystems, both chemically and physically.
For example, molecular oxygen is a vital nutrient for some
microorganisms but a poison to others.
If aerobic (oxygen-consuming) microorganisms remove oxygen
from a habitat, rendering it anoxic (oxygen free), the changed
conditions may favor the growth of anaerobic microorganisms
that were formerly present in the habitat but unable to grow.
In other words, as resources and conditions change in a

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microbial habitat, cell populations rise and fall, changing the

habitat once again.
5. Influence human society in many ways
Microorganisms are closely associated with the health and
welfare of human beings; some microorganisms are beneficial,
and others are detrimental.
Microorganisms are involved in the making of yogurt, cheese and
wine;
In the production of penicillin, interferon and alcohol;
Yogurt is a fermented milk product that contains the
characteristic bacterial cultures Lactobacillus
bulgaricus and Streptococcus thermophilus.
The main (starter) cultures in yogurt are Lactobacillus
bulgaricus and Streptococcus thermophilus.
The function of the starter cultures is to ferment lactose (milk
sugar) to produce lactic acid.
The increase in lactic acid decreases pH and causes the milk to
clot or form the soft gel that is characteristic of yogurt.
The fermentation of lactose also produces the flavor compounds
that are characteristic of yogurt.
Lactobacillus bulgaricus and Streptococcus thermophilus are the
only 2 cultures required by law (CFR) to be present in yogurt.
Other bacterial cultures, such as Lactobacillus
acidophilus, Lactobacillus subsp. casei, and Bifido-bacteria may
be added to yogurt as probiotic cultures.
Probiotic cultures benefit human health by improving lactose
digestion, gastrointestinal function, and stimulating the immune
system.
Detrimental: Microorganisms can cause diseases, spoil food and
deteriorate materials like iron pipes, glass lenses, and wood
pilings.
Importance of microorganisms
 Microbiology is at the center of many important aspects of human and
veterinary medicine, agriculture, and industry.
 For example, although animal and plant infectious diseases are typically
microbial, many microorganisms are essential to soil fertility and
domestic animal welfare.
 Some microorganisms produce biofuels.
 Many large-scale industrial processes, such as the production of

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antibiotics and human proteins, rely heavily on microorganisms.

 Natural gas (methane) is a product of the anaerobic degradation of
organic matter by methanogenic microorganisms
 Ethyl alcohol (ethanol), which is produced by the microbial
fermentation
Level I Institutionally Accredited of glucose from feedstocks such as sugarcane or
cornstarch, is a major motor fuel in some countries
 Waste materials such as domestic refuse, animal wastes, and cellulose
can also be converted to biofuels by microbial activities and are more
efficient feedstocks for ethanol production than is corn.
 Thus, microorganisms affect the everyday lives of humans in both
beneficial and detrimental ways.
Basic Microbiology
 Virology-Viruses and subviral particles
 Microbial physiology - Nutrition, metabolism
 Microbial genetics - Genes, heredity, and genetic variation
 Microbial biochemistry - Enzymes and chemical reactions in cells
 Microbial systematics - Classification and nomenclature
 Molecular biology - Nucleic acids and protein
 Microbial ecology-Microbial diversity and activity in natural habitats;
biogeochemistry
Applied Microbiology
 Medical microbiology - Causative agents of disease; diagnostic
procedures; diagnostic procedures for identification of causative agents;
preventive measures
 Immunology – immune systems
 Biotechnology – production of human proteins by genetically

engineered microorganisms
 Food and dairy microbiology - Food preservation and preparation;

foodborne diseases and their prevention
 Industrial microbiology - Production of medicinal products such as

antibiotics and vaccines; fermented beverages, industrial chemicals;
production of proteins and hormones by genetically engineered
microorganisms; large scale production

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 Agricultural microbiology - Soil fertility; plant and animal diseases
 Plant microbiology and plant pathology – The study of the

interactions
ISO 9001:2015 Certified between microorganisms and plants and plant pathogens.
 Soil microbiology – The study of those microorganisms that are found

in soil.
 Veterinary microbiology – The study of the role in microbes in

veterinary medicine or animal taxonomy.
 Environmental microbiology – The study of the function and diversity

of microbes in their natural environments. This involves the
characterization of key bacterial habitats such as the rhizosphere and
phyllosphere, soil and groundwater ecosystems, open oceans or
extreme environments (extremophiles). This field includes other
branches of microbiology such as: microbial ecology (microbially-
mediated nutrient cycling), geomicrobiology, (microbial diversity),
water microbiology (the study of those microorganisms that are found
in water), aeromicrobiology (the study of airborne microorganisms)
and epidemiology (the study of the incidence, spread, and control of
disease
 Aquatic microbiology – microbial processes in waters and

wastewaters, drinking water safety
Immunization, Antiseptics, and Antibiotics

 Understanding microbes gives us the ability to fight pathogens using
immunization, antiseptics, and antibiotics.
 Surprisingly, most microbes are not harmful to humans.
 In fact, they are all around us and even a part of us.
 However, some microbes are human pathogens; to combat these, we
use immunization, antiseptics, and antibiotics.
 Immunization is the process by which an individual’s immune system
becomes fortified against an agent (known as the immunogen)
 Immunization is the process by which an individual is made immune or
resistant to an infectious disease.
 Vaccines stimulates the body’s own immune system to protect the
person against subsequent infection or disease.
 When the immune system is exposed to molecules that are foreign to
the body, it will orchestrate an immune response

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 It will also develop the ability to respond quickly to subsequent

encounters with the same substance, a phenomenon known as
immunological memory.
 Therefore, by exposing a person to an immunogen in a controlled way,
the body
Level I Institutionally can learn to protect itself: this is called active immunization.
Accredited
 Vaccines against microorganisms that cause diseases can prepare the

body’s immune system, thus helping it fight or prevent an infection.
 The most important elements of the immune system that are improved
by immunization are the T cells, the B cells, and the antibodies B cells
produce.
 Memory B cells and memory T cells are responsible for the swift
response to a second encounter with a foreign molecule.
 By immunizations, some infections and diseases have been almost
completely eradicated throughout the United States and the world.
 For example, polio was eliminated in the U.S. in 1979.
 Active immunization and vaccination have been named one of the “Ten
Great Public Health Achievements in the 20th Century.”
 Antiseptics are antimicrobial substances that are applied to living
tissue to reduce the possibility of infection, sepsis, or putrefaction.
 Their earliest known systematic use was in the ancient practice of
embalming the dead.
 Some antiseptics are true germicides, capable of destroying microbes
(bactericidal), while others are bacteriostatic and only prevent or
inhibit bacterial growth.
 Antisepsis is the process of reducing the number of microorganisms on
skin, mucous membranes, or other body tissue by applying an
antimicrobial (antiseptic) agent.
 Antibiotic is a drug that fights infections caused by bacteria in both
humans and animals.
 Antibiotics fight these infections either by killing the bacteria or making
it difficult for the bacteria to grow and multiply.
 Antibiotics do not have any effect on viruses or fungi. Microbicides that
destroy virus particles are called viricides or antivirals
 The word “antibiotic” was first used in 1942 by Selman Waksman and
his collaborators to describe any substance produced by a
microorganism that is antagonistic to the growth of other
microorganisms in high dilution.
 With advances in medicinal chemistry, most of today’s antibacterial are

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semisynthetic modifications of various natural compounds.

 Resident flora are microorganisms that live in the deeper layers of the
skin and within hair follicles and cannot be completely removed, even
by vigorous washing and rinsing with plain soap and clean water.
Level I In most
Institutionally cases, resident flora is not likely to be associated with
Accredited
infections; however, the hands or fingernails of some health care

workers (HCWs) can become colonized by microorganisms that do
cause infection (e.g., Staphylococcus aureus, gram-negative bacilli, or
yeast), which can be transmitted to patients.
 Transient flora are microorganisms acquired through contact with
individuals or contaminated surfaces during normal, daily activities.
 They live in the upper layers of the skin and are more amenable to
removal by hand hygiene.
 They are the microorganisms most likely to cause health care-associated
infections
The future of microbiology
 infectious diseases
 new and improved industrial processes
 microbial diversity and microbial ecology
History of Microbiology

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© Art Collection 3 / Alamy Stock Photo image ID:

HYAYHE
A drawing of the microscope used by Robert Hooke in 1664. The lens was fitted at
the end of an adjustable bellows (G) and light focused on the specimen by a
separate lens (1

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image credit:
imagefamousscientists.org/robert-hooke/
credit: FIGURE 4.2 https://schoolbag.info/biology/concepts/19.html
This drawing of a mold that was growing on
the surface of leather, together with other
drawings and accompanying text published The Van Leeuwenhoek microscope. A replica
by Robert Hooke in Micrographia in 1665, of Antoni van Leeuwenhoek’s microscope.
1673-1723, Antoni van Leeuwenhoek (Dutch)
were the first descriptions of microorganisms.
described live microorganisms that he
The round structures contain spores of the
observed in teeth scrapings, rainwater, and
mold.
peppercorn infusions.
Copyright © 2009 Pearson Education Inc publishing as Copyright ©: 2009 Pearson Education Inc publishing as
Benjamin Cummings Benjamin Cummings
Leeuwenhoek’s drawings of bacteria Photomicrograph of a human blood smear

published in 1684. Even from these taken through a van Leeuwenhoek
simple drawings we can recognize microscope. Red blood cells are clearly
several shapes of common bacteria: A, C, apparent.
F, and G, rods; E, cocci; H, packets of
cocci.
Drawing by Ferdinand Cohn of large

filamentous sulfur-oxidizing bacteria
Beggiatoa mirabilis. The small
granules inside the cells consist of
elemental sulfur, produced from the

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Copyright © 2009 Pearson Education Inc publishing as Benjamin Cummings
 Cohn was the first to identify the granules as sulfur in 1866.

Level I A cellAccredited
Institutionally of B. mirabilis is about 15 m in diameter.
 Beggiatoa moves on solid surfaces by a gliding mechanism and in so
doing, cells often twist about one another.
 Cohn is credited with many other accomplishments.
 He laid the groundwork for a system of bacterial classification,
including an early attempt to define a bacterial species, an issue still
unresolved today, and founded a major scientific journal of plant and
microbial biology.
 He strongly advocated use of the techniques and research of Robert
Koch, the first medical microbiologist. Cohn devised simple but effective
methods for preventing the contamination of culture media, such as the
use of cotton for closing flasks and tubes.
 These methods were later used by Koch and allowed him to make rapid
progress in the isolation and characterization of several disease-causing
bacteria.
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The Golden Age of Microbiology 1857-1914
Beginning with Pasteur’s work, discoveries included the relationship

between microbes and disease, immunity, and antimicrobial drugs
The Germ Theory of Disease
 1835: Agostino Bassi showed a silkworm disease was caused by a
fungus.
 1865: Pasteur believed that another silkworm disease was caused by a
protozoan.

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 1840s: Ignaz Semmelwise advocated handwashing to prevent

transmission of puerperal fever from one patient to another.
 1860s: Joseph Lister used a chemical disinfectant to prevent surgical
wound infections after looking at Pasteur’s work showing microbes are
in theAccredited
Level I Institutionally air, can spoil food, and cause animal diseases.
 1876: Robert Koch provided proof that a bacterium causes anthrax and
provided the experimental steps, Koch’s postulates, used to prove that a
specific microbe causes a specific disease. Koch was a physician and
Pasteur’s young rival
 A young milkmaid informed the physician Edward Jenner that she could
not get smallpox because she had already been sick from cowpox.
 1796: Edward Jenner inoculated a person with cowpox virus. The
person was then protected from smallpox. Called vaccination from
vacca for cow. The protection is called immunity
Vaccinations
 produced from avirulent microbial strains
 produced from live viruses
 produced from viral particles
cont….(history)
 1910: Paul Ehrlich developed a synthetic arsenic drug, salvarsan, to
treat syphilis.
 1930s: Sulfonamides were synthesized.
 1928: Alexander Fleming discovered the first antibiotic. He observed
that Penicillium fungus made an antibiotic, penicillin, that killed S.
aureus.
 1940s: Penicillin was tested clinically, and mass produced.
Classification of Microbes
Taxonomy
 The science of classifying organisms
 Provides universal names for organisms
 Provides a reference for identifying organisms
Copyright ©: 2004 Pearson Education Inc, publishing as Benjamin Cummings

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Taxonomic Hierarchy
Domain
Kingdom
Phylum
Level I Institutionally Accredited Class
Order
Family
Genus
Species
Classification of Microbes
 Eukaryotic species:
A group of closely related organisms that breed among
themselves
Eukaryotic microbes are an extraordinarily diverse group,
including species with a wide range of life cycles, morphological
specializations, and nutritional needs.
Although more diseases are caused by viruses and bacteria than

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by microscopic eukaryotes, these eukaryotes are responsible for

some diseases of great public health importance.
Eukaryotes include such microorganisms as fungi, protozoa, and
simple algae
Level I Prokaryotic
Institutionally Accredited species:
Microbes within the domains Bacteria and Archaea are all

prokaryotes (their cells lack a nucleus)
Bacteria are found in nearly every habitat on earth, including
within and on humans.
Most bacteria are harmless or helpful, but some are pathogens,
causing disease in humans and other animals.
 Viral species:
Viruses are the smallest of all the microbes.
Viruses are acellular (not composed of cells).
Prokaryotic species:
Bacteria (or Eubacteria)

 Most abundant on earth
 They are nitrogen fixers and recycle carbon
 No membrane bound organelles
Archaea
 The entire subclass of archaea are also prokaryotes, mostly remarkable
because of their ability to thrive in very harsh environments. An
example of archaea is the Sulfolobus acidocaldarius archeobacterium
that lives in extremely acidic mud pots in geothermally active areas.
 Methanogens
 Halophiles
 Hyperthermophiles
© Life Science Clipart - My Adventure to The Science Center
Example of some microbes

Prokaryotes Eukayotes
Escherichia coli Paramecium caudatum
Salmonella Euglena acus
Streptococcus Saccharomyces
 S. pneumoniae  Saccharomyces cerevisiae
 S. pyogenes
 S. mutans

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Streptomyces Aspergillus
 Streptomyces griseus  Aspergillus fumigatus
 S. aureofaciens
 S. rimosis
 S. erythraeus
 S. venezuelae
Helicobacter pylori Penicillium marneffei

Staphylococcus aureus Neurospora
 Neurospora crassa
 Neurospora sitophila
 Neurospora intermedia
 N. tetrasperma and N. discreta
Methanobrevibacter smithii
Halobacterium salinarum
Thermococcus litoralis
Sulfolobus acidocaldarius
Microbes and Human Disease

 Bacteria were once classified as plants which gave rise to use of the
term flora for microbes.
 This term has been replaced by microbiota.
 Microbes normally present in and on the human body are called normal
microbiota.
 Normal microbiota prevents growth of pathogens.
 Normal microbiota produces growth factors such as folic acid and
vitamin K.

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 Resistance is the ability of the body to ward off disease.

 Resistance factors include skin, stomach acid, and antimicrobial
chemicals.
 When a pathogen overcomes the host’s resistance, disease results.
Level I Emerging
Institutionally Accredited Infectious Diseases (EID): New diseases and diseases
increasing in incidence
 Multidrug Resistance (MDR) - When a single bacterium is resistant to
more than one antibiotic
Major Taxonomic Groups of Bacteria per Bergey’s manual
 Gracilicutes – gram-negative cell walls, thin-skinned
 Firmicutes – gram-positive cell walls, thick skinned
 Tenericutes – lack a cell wall & are soft
 Mendosicutes – archaea, primitive procaryotes with unusual cell walls
& nutritional habits
MICROBIOLOGY LABORATORY: BASIC RULES AND REQUIREMENTS
Microbiology Lab Practices and Safety Rules
 Wash your hands with disinfectants when you arrive at the lab and
again before you leave.
 Wear laboratory coats in the lab. Students with long hair must put up
the hair.
 At the start and end of each laboratory session, students should
clean their assigned bench-top area with a disinfectant solution

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provided. That space should then be kept neat, clean, and

uncluttered throughout each laboratory period.
 Eating or drinking in the laboratory is not permitted. No mouth
ISO 9001:2015 pipetting.
Certified
 Label everything clearly. Sterilize equipment and materials.

 Avoid loose fitting items of clothing. Wear appropriate shoes in the
laboratory.
 Report any breakage of equipment to the instructor.
 Report any personal accidents such as cuts to the instructor at once.
 Turn off Bunsen burner when not in use.
 Discard all cultures and used glassware into the container labeled
CONTAMINATED. (This container will later be sterilized.) Plastic or
other disposable items should be discarded separately from
glassware in containers to be sterilized.
 Never place contaminated pipettes on the bench top.
 When you flame sterilize with alcohol, be sure that you do not have any
papers under you.
 Before beginning your laboratory exercise, wash off the bench top
with the disinfectant provided. When exercises are completed, wash
off the bench top again. Always wash your hands with soap and
water before leaving the laboratory.
 Before leaving the laboratory, see that all the equipments are in the
proper location and gas and water turned off.
 Purchase a fine point, waterproof marker and small roll of masking
tape. Use them to clearly label your cultures.
 If you should spill or drop a culture or if any type of accident occurs,
call the instructor immediately. Place a paper towel over any spill
and pour disinfectant over the towel. Let the disinfectant stand for
15 minutes and then clean the spill with fresh paper towels.
Remember to discard the paper towels in the proper receptacle and
wash your hands carefully.
 Disinfect work areas before and after use with 70% alcohol or fresh
10% bleach. Laboratory equipment and work surfaces should be
decontaminated with an appropriate disinfectant on a routine basis
and especially after spills, splashes or other contamination.
 Replace caps on reagents, solution bottles and bacterial cultures. Do
not open petri dishes in the lab unless absolutely necessary.
 Cultures are not to be removed from the laboratory unless the
instructor gives permission.

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 Always place culture tubes (broth and slants) in the upright position
in a rack or basket for incubation or disposal.
 Dispose off all solid waste materials in a biohazard bag and
autoclave it before discarding in the regular trash.
Level I TreatAccredited
Institutionally all cultures as potentially pathogenic, i.e., flood areas with
disinfectant if cultures are spilled, wash hands after contact and
notify your instructor at once.
 Read the instructions carefully before beginning an exercise. Also,
make sure you have all the materials needed for the exercise at hand
before you commence the experiment. Ask the instructor for
clarification of any points about which you are in doubt.
 Flame the inoculating loop or needle immediately before and after
use. If viscous material is present on the loop or needle, dry it at the
side of the flame before placing it directly in the flame.
 Laboratory notebooks must be kept up to date. Illustrations should be
done when requested.
 Make sure you consult the instructor to dispose of the cultures that
are not needed any longer. Remove all labels and markings from the
tubes before disposing of them; do not discard anything into the
sinks.
 Please inform your instructor if you have any medical condition that
could potentially affect your safety in the laboratory (eg: diabetes,
epilepsy, immunosuppression etc.). This information will help the
instructor to deal with any emergency that would arise. The
information will be treated confidentially, and it will not affect their
ability to participate in the laboratory activities.
 Be systematic and logical. Keep a faithful record of all the
experiments and observations. Update it regularly and submit it for
evaluation at the end of each exercise.
 Work either using laminar air flow chamber or light the burner at
least five minutes prior to making any inoculations and work near
the burner.
Basic requirements of a microbiology laboratory
A microbiology laboratory requires well-built rooms equipped with

glassware, tools and equipment. Some of the most important items
of equipment are the following.

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Common Glassware
The most important glassware used in a microbiological laboratory

are test tubes, culture tubes, Petri dishes, Measuring cylinder,
pipettes,
Level I Institutionally Accreditedglass spreader, Flasks, screw-capped glass bottles,
haemocytometer etc.
 Test Tubes, Culture Tubes and Screw-Capped Tubes

 These are made up of glass, one end of which is closed,
and another end is open.
 If the side wall of the open end is slightly curved
outside, it is called test tube; if the side wall is smooth,
it is called culture tube. When the side wall of the tube
has screws so that a plastic cap may be fitted, it is
called screw-capped tube.
 The test tubes are used for testing the chemicals such
as pH etc., culture tubes are used for preparation of
agar slants and purification of microorganisms. The
open end is plugged with non-absorbent cotton plug.
 Sometimes the microorganisms are purified and
preserved in screw-capped tubes.
 Petri dish
R. J. Petri, a student of the most renowned bacteriologist Robert
Koch devised this dish, hence called “Petri dish”.
It consists of two shallow glass dishes, the upper half or lid and the
lower half.
For the isolation and cultivation of different types of
microorganisms these dishes are used in all microbiological
laboratories.
According to the requirement, its diameter varies.
Molten agar medium is aseptically poured on the lower half of
the sterilized Petri dish and then covered with the upper half.
The petri dishes are sterilized by putting them in a Petri dish
container and in turn in an oven or autoclave.
 Pipette
It is a cylindrical and graduated glass apparatus.
It’s one end (lower side) tapers, while the other end
(mouthpiece) is normal. The middle portion is wider or of the

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same size as mouth end.

It is graduated with numbers 1, 2 . . . n.....10.
It has different measuring capacity such as 0.1, 0.5, 1, 5, 10 ml etc.
Hence measures different quantities.
It is used for transferring appropriate amount of liquid into other

containers.
It should be sterilized in an oven or autoclave before use by
keeping in pipette container after being plugged with cotton.
For safety point, liquid should be sucked by attaching pipette-sucker
at the normal end of the pipette.
Pipettes should be sterilized by keeping them first in a steel can and
is sterilized at 121°C for 30 minutes
 Glass Spreader
It is made by bending a glass rod and making an L-shaped structure.
It is used to spread evenly the microorganisms on agar surface
present in liquid medium.
The long arm is held in hand and the small arm is flame-sterilized
and put on agar surface.
It is brought forth and back so that microorganisms present in
liquid may be dissociated and evenly spread on the entire
surface of agar.
 Haemocytometer
This is a device used to measure the blood cells.
This is also used for counting other cells viz., spores, bacteria etc.
It consists of a number of chambers. Each big chamber has 1 X 1
X 0.1 mm = 0.1mm3 volume with an area of 1 X 1mm = 1mm 2.
The depth of chamber is 0.1mm. (1 X 1 X 0.1 mm = 0.1mm 3 =
0.0001cm3 = 10-4cm3 = 10-4 ml) Hence, the bacterial cell count in
the large chamber will be multiplied by 10 4 to give an estimate
of bacterial cell number/ml.
Each large chamber has 9 medium-sized chambers with 0.2 mm
length, 0.2 mm width and 0.1mm depth with a volume of 0.004
mm3.
Each medium sized chamber is divided into 25 small chambers
with 0.04 mm length, 0.04 mm width and 0.1mm depth with a
volume of 0.00016 mm3.
 Cleaning of Glass wares. Care should be taken that the glassware

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used in microbiology laboratory are neat and clean. After receiving

and before start of work, the glassware must be chemically cleaned
so that there should not be chemical deposits on its surface.
Moreover, dirty form, then by soaking overnight in chromic acid. If
the latter
Level I Institutionally is not effective, a mixture of concentrated nitric acid and
Accredited
sulphuric acid can be applied. All traces of the cleaner are then
removed by repeated rinsing in tap water followed by distilled
water. Sometimes, new glassware may contain some
bacterial/fungal spores that come with packing materials. The
glassware from factory also contains some amount of alkali, hence,
to remove the alkali, 2-3 % HCl is applied for 24 hours for
neutralization.
After the use of glassware, the medium is removed, and the

former is treated with 3% commercially available Lysol solution
followed by washing with boiling water. After drying, it can be
kept in oven for 3-6 hours at 140-180°C. The used pipettes,
slides, cover slips, petri dishes, etc. are also cleaned by this
method.
Chromic acid is widely used as cleaning agent for glassware. It is

a mixture of sodium dichromate and concentrated sulfuric acid
and possesses powerful oxidizing and solvent properties.
Preparation of Chromic acid
 First Method: Weigh 5g of sodium potassium dichromate and

dissolve in 5ml distilled water in a beaker (250ml). Add 100ml
concentrated H2SO4 slowly and stirring it constantly. The mixture is
allowed to cool at about 40°C and then stored in a dry glass
stoppered bottle.
 Second Method: Weigh 1g of K2Cr2O7 or Na2Cr2O7 and mix with
100ml of concentrated H2SO4. After stirring several times
(preventing from cake formation) allow the mixture to cool at 40°C
and store in a clean, dry stoppered bottle.
Tools in Microbiology Laboratory

The most common equipment are inoculation needle,
inoculation/transfer loop, Bunsen burner, autoclave (or pressure

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cooker), incubator, hot air oven, refrigerator, centrifuge,

spectrophotometer, magnetic stirrer, orbital shaker, hot plate,
Distillation water still, UV- lamp, water-bath, carbon dioxide
cylinder, single-pan balance with weights (for rough use), chemical
balance,
Level I Institutionally pH meter, colony counter, laminar air flow, electrophoretic
Accredited
apparatus, microscopes etc.
Inoculation Needle & Inoculation Loop

These are the most used tools.
Inoculation needle/loop is made up of a long platinum wire fixed
into a metallic rod.
A wire loop has a handle with steel screw shaft in which nichrome
or platinum wire is to be fitted.
The straight wire needle is used for transferring culture from
solid medium. Even smaller amount of liquid culture can be
manipulated by using straight needle.
The loop and wire are also used for picking small quantities of
solid materials from a microbial colony and can be used to
inoculate either a liquid or a solid medium. Both the loop and
straight wire must be flamed immediately after use to avoid
contamination.
Bunsen Burner
Sterilization of tools by using spirit lamp is called incineration.
Gas enters the burner at the base, and its supply is regulated
externally by the gas cock.
The amount of air can be controlled by rotating a sleeve that fits
over the holes in the body of the burner.
To keep the flame from blowing out special tips are frequently used
to fit over the top end of the barrel.
The proper method of lighting the burner is to close off the air
supply, turn on the gas and light. The flame will be large and
yellow. Gradually open the air intake until the flame takes a blue
colour.
Water Bath
Water bath is an instrument that is used to provide constant
temperature to a sample.
It consists of an insulating box made up of steel fitted with electrode
heating coil.

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The temperature is controlled through a thermostat.

In some of the water baths, plate form rotates, then it is called
water bath shaker. It is more useful to microbiologists because it
provides a uniform heat to the sample material meant for
incubation.
The main use of water bath is the incubation of samples at a desired
and constant temperature.
Laminar Air Flow Chamber

Laminar air flow is an apparatus consists of an air blower in the
rear side of the chamber which can produce air flow with
uniform velocity along parallel flow lines. There is a special filter
system of high efficiency particulate air filter (HEPA) which can
remove particles as small as 0.3 mm.
In front of the blower, there lies a mechanism through which air
blown from the blower produces air velocity along parallel flow
lines.
The laminar air flow is based on flow of air current of uniform
velocity along parallel flow lines which help in transferring
microbial cultures in aseptic conditions. Air is passed through
the filters into the enclosure and the filters do not allow any
kind of microbe to enter the system.
Inside the chamber one fluorescent tube and another UV tube
are fitted. Two switches for these tubes and a separate switch
for regulation of the air flow are fitted outside the LAF. Due to
uniform velocity and parallel flow of air current, pouring of
media, plating, slant preparations, streaking etc. are performed
without any kind of contamination.
Initially, dust particles are removed from the surface of the
laminar air flow with the help of smooth cloth containing
alcohol. Switch on the UV light for a period of 30 minutes so as
to kill the germs, if any present in the area of working space.
The front cover sheet of the apparatus is opened to keep the
desired material inside. The air blower is set at the desired
degree so that the air inside the chamber is expelled because the
air inside the chamber may be contaminated / bring
contaminants.
Sit properly in front of the chamber and wipe the working table
with alcohol to reduce the contaminants. All the work related to

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pouring, plating, streaking etc. are to be carried out in the flame

zone of the burner or spirit lamp.
In microbiology laboratory, horizontal type of laminar air flow is
used to supply the air through filter.

Precautions: Put off the shoes before entering to operate the
apparatus. Wash the hands with detergents or soap. One should not
talk inside the chamber while performing microbial culture
transfer, failing which chances of contamination may be more
which may come either through mouth, sneezing or air.
Incubator
An incubator is an instrument that consists of copper/steel
chamber around which warm water or air is circulated by
electric current or by means of small gas flame.
The temperature of the incubator is kept constant due to its control
by using thermostat.
The incubator is made up of double walled chamber adjusted to
a desired temperature. It is done by using an external knob
controlling the thermostat system. The gap between two walls is
insulated to check heat condition. A thermometer is inserted
from the top for recording the temperature.
Temperature greatly influences the microbial growth. Therefore,
instrument is generally designed that can allow the desired
microorganism to grow at a temperature.
It is operated to allow the microbial growth on a suitable
medium under proper temperature. In an incubator, the
variation in temperature should not be more than one degree.
Small square type incubators are better than large ones. If a
lower temperature than the room is required, the water is
circulated around the chamber to pass through an ice chest.
Precautions: the door of the incubator should be opened only

when necessary. If the tubes are to be incubated for a long time or
at higher temperature, the medium may become too dry due to
excessive evaporation. In such cases cotton plug should be pushed
inside the neck of the tube. The tube should be covered by a rubber
cap to cover the plug. If the petriplates are to be incubated for a

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long time, they may be placed in moist chamber with a damp sterile
cotton wool at the bottom.
Colony Counter
It is a device used for counting small or closely growing colonies on

the surface of media.
For accuracy, the lens fitted or mounted in it helps to see the
colonies.
The lens is movable on the box and can be adjusted to see the
colonies.
The petriplate is kept on a slanting platform meant for it and
illuminated with the help of light source from beneath.
The numbers of colonies are read out with the support of digital
reading meter.

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Inoculation Loop & Needle Bunsen Burner Water Bath
Laminar Air Flow
Colony Counter
Incubator
Fig. 1. Tools in microbiology laboratory
QUIZ

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Harmful Microorganisms Sorting. This allow students to examine

multiple microorganisms (microbes) and decide what type of microbes
caused
ISO 9001:2015 Certified that disease.
Further details to be posted on Friday, October 16, 2020 at 8:00 am

(Google Classroom)
Disease caused by Disease caused by virus Disease caused by

bacteria fungi/protists
Performance Tasks
\Homework:
Instructions: Listen to the “Rodney vs. Death” Radiolab Podcast. Click
https://www.wnycstudios.org/podcasts/radiolab/articles/312245-rodney-versus-death and listen to the podcast
then answer the following questions below Your grade will be based on how complete and correct your answers
are.
1. This podcast begins with the story of a teenage girl, named Jeanna, who developed mysterious medical
symptoms. Summarize the beginning of her illness, before she went to see the infectious disease expert in
Milwaukee. What significant piece of information led Jeanna’s pediatrician to refer her to an infectious disease
expert?
2. Once Jeanna was transferred to Dr. Willoughby of Milwaukee Children’s hospital, how did her symptoms progress?
3. Throughout history rabies has been a disease that is essentially 100% fatal. So how did the crazy historical “cures”

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described, such as application of a rooster’s anus to the bite wound, ever catch on?
4. How does the rabies virus enter the body and then travel to the brain, its target tissue? How is this different than
how most viruses find their target
ISO 9001:2015 tissue in the body?
Certified

5. If you are bitten by a rabid animal, you can still avoid developing rabies if you get vaccinated. At what point is it too
late for the vaccine to help?
6. What are the symptoms of rabies? Describe the course of the disease. Why would the symptom of hydrophobia
developed by rabies victims be advantageous for the virus?
7. What is excitotoxicity and what might it have to do with how a rabies infection affects the brain?
8. What unconventional treatment did Dr. Willoughby try with Jeanna and how does it relate to excitotoxicity in the
brain? Why did the doctor think that this untested treatment might work? What was his rationale? And finally, did
it work?
9. State the reason why majority of the doctors are abandoning and not recommending Milwaukee Protocol?
10. Jeanna’s treatment by Dr. Willoughby is now referred to at the Milwaukee Protocol. What is its current success
rate?
This assignment is adapted from the Virtual Microbiology Classroom (http://www.scienceprofonline.com/virtual-micro-main.html) on the free science education
website Science Prof Online (ScienceProfOnline.com). Visit the website to find more science education resources such as lecture PowerPoints, practice test
questions, review questions, science photos, videos and assignments.
Understanding Directed Assess
Rubric for Microorganisms Sorting

• Every correct answer is equivalent to 2 points
• Total of 60 points
Rubric for Homework
• Your grade will be based on how complete and correct your answers are.
• Total of 50 points

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Learning Resources

Madigan, M., Martinko, J., Stahl, D., and Clark, D. 2012. Brock Microbiology of
Microorganisms 13th Accredited
Level I Institutionally edition. Benjamin Cummings, USA.
Varghese, N., Joy, PP. (2014, January). Microbiology Laboratory Manual. Vazhakulam, Muvattupuzha,
Ernakulam District, Kerala, PIN-686 670
Port, Tami. (2016). Virtual Microbiology Classroom. Retrieved from Virtual Microbiology Classroom
(http://www.scienceprofonline.com/virtual-micro-main.html
Kaiser, Gary. (2020). Introduction to Microbiology. Retrieved from

https://bio.libretexts.org/Bookshelves/Microbiology/Book
%3A_Microbiology_(Kaiser)/Unit_1%3A_Introduction_to_Microbiology_and_Prokaryotic_C
ell_Anatomy/1%3A_Fundamentals_of_Microbiology/1.1%3A_Introduction_to_Microbiolog
y
Intellectual Property
This module is for educational purpose only. Under section Sec. 185 of RA 8293, which states, “The
fair use of a copyrighted work for criticism, comment, news reporting, teaching including multiple
copies for classroom use, scholarship, research, and similar purposes is not an infringement of
copyright.”
The unauthorized reproduction, use, and dissemination of this module without joint consent of the
authors is strictly prohibited and shall be prosecuted to the full extent of the law, including
appropriate administrative sanctions, civil, and criminal.

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Republic of the Philippines

Laguna State Polytechnic University

LSPU Self-Paced Learning Module (SLM)

Level I Institutionally Accredited

Student Learning Strategies

Online Activities A. Online Discussion via Zoom

The online discussion will happen on October 12 or 13 2020, 10 am-12

B. Learning Guide Questions:

Offline Activities Lecture Guide

LSPU SELF-PACED LEARNING MODULE: TECHNOLOGY FOR TEACHING AND LEARNING

and waters, in the human body, and in animals and plants.

The Science of Microbiology

1. First living organisms on planet

LSPU SELF-PACED LEARNING MODULE: TECHNOLOGY FOR TEACHING AND LEARNING

The cyanobacteria had given importance in biological evolution

LSPU SELF-PACED LEARNING MODULE: TECHNOLOGY FOR TEACHING AND LEARNING

microbial habitat, cell populations rise and fall, changing the

LSPU SELF-PACED LEARNING MODULE: TECHNOLOGY FOR TEACHING AND LEARNING

antibiotics and human proteins, rely heavily on microorganisms.

 Immunology – immune systems

 Biotechnology – production of human proteins by genetically

 Food and dairy microbiology - Food preservation and preparation;

 Industrial microbiology - Production of medicinal products such as

LSPU SELF-PACED LEARNING MODULE: TECHNOLOGY FOR TEACHING AND LEARNING

 Agricultural microbiology - Soil fertility; plant and animal diseases

 Plant microbiology and plant pathology – The study of the

 Soil microbiology – The study of those microorganisms that are found

 Veterinary microbiology – The study of the role in microbes in

 Environmental microbiology – The study of the function and diversity

 Aquatic microbiology – microbial processes in waters and

Immunization, Antiseptics, and Antibiotics

LSPU SELF-PACED LEARNING MODULE: TECHNOLOGY FOR TEACHING AND LEARNING

 It will also develop the ability to respond quickly to subsequent

 Vaccines against microorganisms that cause diseases can prepare the

LSPU SELF-PACED LEARNING MODULE: TECHNOLOGY FOR TEACHING AND LEARNING

semisynthetic modifications of various natural compounds.

infections; however, the hands or fingernails of some health care

LSPU SELF-PACED LEARNING MODULE: TECHNOLOGY FOR TEACHING AND LEARNING

ISO 9001:2015 Certified

Level I Institutionally Accredited

© Art Collection 3 / Alamy Stock Photo image ID:

LSPU SELF-PACED LEARNING MODULE: TECHNOLOGY FOR TEACHING AND LEARNING

ISO 9001:2015 Certified

Level I Institutionally Accredited

Leeuwenhoek’s drawings of bacteria Photomicrograph of a human blood smear

Drawing by Ferdinand Cohn of large

LSPU SELF-PACED LEARNING MODULE: TECHNOLOGY FOR TEACHING AND LEARNING

Copyright © 2009 Pearson Education Inc publishing as Benjamin Cummings

 Cohn was the ﬁrst to identify the granules as sulfur in 1866.

The Golden Age of Microbiology 1857-1914

Beginning with Pasteur’s work, discoveries included the relationship

LSPU SELF-PACED LEARNING MODULE: TECHNOLOGY FOR TEACHING AND LEARNING

 1840s: Ignaz Semmelwise advocated handwashing to prevent

Copyright ©: 2004 Pearson Education Inc, publishing as Benjamin Cummings

LSPU SELF-PACED LEARNING MODULE: TECHNOLOGY FOR TEACHING AND LEARNING

LSPU SELF-PACED LEARNING MODULE: TECHNOLOGY FOR TEACHING AND LEARNING

by microscopic eukaryotes, these eukaryotes are responsible for

Microbes within the domains Bacteria and Archaea are all

Bacteria (or Eubacteria)

Example of some microbes

LSPU SELF-PACED LEARNING MODULE: TECHNOLOGY FOR TEACHING AND LEARNING

Helicobacter pylori Penicillium marneffei

Microbes and Human Disease

LSPU SELF-PACED LEARNING MODULE: TECHNOLOGY FOR TEACHING AND LEARNING