Microbiology

(Author : Umossoh A. A.
CHAPTER 1
1.1 INTRODUCTION TO MICROBIOLOGY

Microbiology can be defined as a branch of science that deals with the study of
microscopic entities called microorganisms which includes bacteria, archaea, some
fungi, protozoa and viruses.
More so, it is concerned with the structure, function and classification of
microorganisms coupled with ways of both exploiting and controlling their activities.
Microbiology encompasses many discipline such as
● Virology - Study of viruses, its structures, functions and classification.
● Mycology - Study of fungi, its structures, functions and classification.
● Bacteriology - Study of bacteria, its structures, functions and

classification.
● Parasitology - Study of parasites, their host and relationship between
them.
● Public Health & Epidemiology – Study of diseases, its transmission,
distribution and control over a given population.
● Food Microbiology – Study of microorganisms both as food spoilage
agents and those useful in food production.
● Medical Microbiology – Study of the diagnosis, treatment and prevention
of the activity of microorganism in infectious disease.
● Environmental Microbiology – Study of microorganism and its harmful
and beneficial effects on the environment.
● Immunology and Immunochemistry – Study of the immune system and
the impact of the activities of microorganisms on it.
● Petroleum Microbiology – Study of microorganisms that can metabolize
or alter petroleum products.
● Soil and Agricultural Microbiology – Study of the impact of
microorganisms on soil and their effect on agricultural products and
agricultural activities.
● Phycology – Study of algae, its structures, functions and classification.
● Protozoology - Study of protozoa, its structures, functions and

classification.
● Pharmaceutical Microbiology – The use of microorganisms for inhibiting
contamination as well as development of pharmaceuticals.
● Veterinary Microbiology – Deals with the study of microorganisms that
causes diseases in animals.
● Aquatic Microbiology – Study of microorganisms of aquatic system either
fresh or marine water system.
1.2 TERMS IN MICROBIOLOGY

Antiseptic – A chemical antimicrobial agent that kills some or inhibits the growth and
reproduction of microorganisms when applied on the surface of a living tissue.
Antiseptic technique – These are set of procedures observed in a laboratory to prevent cross
contamination of research work.
Bactericidal – A substance of chemical complexity that kills bacteria out-rightly.

Bacteriostatic – A chemical substance that inhibits bacterial growth and reproduction.
Normal microbiota-These are microorganisms that are found living harmlessly mutualistic on
various parts of the body e.g. Staphylococcus sp on skin, Salmonella sp in the gastrointestinal
tract, Lactobacillus in the vagina.
Sterilizer – A physical process or chemical agent applied on inanimate object to kill all forms of
microbial life present on the surface.
Sterilization – A process that kills out-rightly all forms of microbial life and biological agents on
the surface of an inanimate object.
Pathogen – A biological agent that causes diseases when introduced into a susceptible heathy
host.
Disinfection – A process which eliminates most microbial life on a non living tissue or
inanimate or inert surface.
Disinfectant – A substance of chemical complexity that eliminates most microbial life when
applied on a non living tissue or inanimate or inert surface.
Germicide – Any substance that kills germs.

Pasteurization – Process of food treatment (dairy product, vegetables and fruits) with mild
heat to eliminate pathogen and extend shelf life.
Shelf life – Refers to the length of time that a commodity may be stored without reducing in
quality thus becoming unfit for use, consumption or sale.
Expiring date – A specified time after which a consumable product (food or medicine) should
not be used due to its altered quality and tendency to cause harm and damage if consumed.
Best before date (BB)- A specified time after which a consumable products is no longer in its
perfect shape from that date. It may just lose its freshness, taste, aroma or nutrients. It does
not necessarily mean that the product is no longer safe to consume.
Sterile – A state of total absence of microorganism on a surface.

Preservation – Process involved in maintaining a substance without alteration to its vital
chemical or physical characteristics.
Disease – A particular disorder or abnormal condition that induces negative effect on the
structure or function of all or part(s) of the body of an organism and is not due to an immediate
external injury.
Immunity – A state of non-susceptibility of an organism to diseases.

Reservoir – Population of microorganism in a specific environment in which they live, grow,
reproduce and depend on.
Endemic – A disease that exist permanently in a particular region or population e.g. Malaria in
Nigeria
Epidemic – The outbreak of disease in a particular region or population over a certain period
of time e.g. typhoid fever outbreak in USA 1911.
Pandemic – An epidemic of infectious disease that has spread across a large region/continent,
e.g. Ebola disease, Covid-19.
Vector – Any agent that carries and transfers infectious particles into another living organism,
e.g. Mosquito, Tsetse fly.
Infection – The invasion of an organisms’ body tissue by pathogenic agents to multiply and
apparently cause harm.
Pathogenicity- Refers to the ability of an organism to cause disease (i.e. harm the host).
Virulence- Refers to the degree of pathology caused by the organism.
Pathology- Refers to the study and diagnosis of disease through examination of organs,
tissues, bodily fluids, and whole bodies (autopsies).
Contamination – The presence of constituent impurities and unwanted elements on a

material, body or environment.
Decontamination – A process of treatment that renders a body surface safe to handle by

reducing the concentration of microbial contamination.
Vaccine – A biological substance constituted of weakened or killed form of microbes, its toxins
or its surface protein, that when introduced into a living host, stimulates production of
antibodies and induces immunity against one or several diseases.
Antigen – A substance of internal chemical complexity that is easily solubilized such that when
administered, provokes immune response, eliciting antibody production which confers lasting
immunity to the host.
Antibody – A chemical substance produced by the body to combat infectious agents as a result
of response to an antigenic stimulus.
Culture- A collection or population of microbial growth in a nutrient media.

Media- This is a liquid or solid chemically designed compound or mixture, engineered to
support the growth of microorganism.
Broth- This is a liquid chemically designed compound or mixture, engineered to support the
growth of microorganism.
Agar- A solidifying agent usually introduced to solidify liquid nutrient media in the laboratory.
Usually sourced from rhodophyta (red algae).
Inoculation- Process of transferring microbial cells from source to a media.
Inoculum- This is the microbial cell being transferred during Inoculation.
Serial Dilution- A stepwise dilution of microbial sample in a chronological order using a
physiological saline with the aim of reducing microbial concentration resulting in a barest
minimum standard essential for culturing microorganisms in a pure culture.
Colony- A visible mass of microorganisms originating from a single mother cell appearing
genetically alike.
Glass slide- A glass slide is a thin, flat, rectangular piece of glass that is used as a platform for
observing or view specimen in a microscope. A typical glass slide usually measures 25 mm wide
by 75 mm, or 1 inch by 3 inches long, and is designed to fit under the stage clips on a
microscope stage.
Smear- A sample of tissue, cells or other material taken from part of the body, spread thinly on
a microscope glass slide by a loop or swab for microscopic examination, typically for medical
diagnosis.
Recombinant: This refers to something formed by combining existing elements in a new

combination. Thus, the phrase recombinant DNA refers to an organism created in the lab by
adding DNA from another species.
Serum: The clear yellowish fluid obtained upon separating whole blood into its solid and liquid
components after it has been allowed to clot. Also called blood serum.
1.3 CONTRIBUTION OF SCIENTISTS TO MICROBIOLOGY
Antonio Van Leeuwenhoek in the 16th century is considered the “Father of Microbiology”. He
used homemade microscope to discover microbial entities in a drop of water from murky lake,
he later named these microbial entities “animalcules”. He discovered red blood cell, sperm cell
and bacteria not long after that.
Louis Pasteur a French chemist and microbiologist is considered “Father of Modern

Microbiology”. He discovered that microorganism causes fermentation in food product thus
originated the food preservation method called Pasteurization. He also developed vaccine
against anthrax and rabies.
Ferdinand Cohn a German biologist is considered the “Father of Modern Bacteriology”. He
discovered the heat resistant endospores of bacteria.
John Tyndall an Irish physicist discovered that bacteria exist in two forms, ones that are killed
by heat and heat resistant bacteria. He also discovered Tyndallization sterilization; a process for
sterilizing substances usually food substances that kills heat resistant endospores.
Robert Koch a German Doctor discovered causal organism of anthrax, tuberculosis, cholera
and typhoid. He also developed an easier way to observe bacteria using the Petri dish, and
developed the staining technique. In 1884, he developed a guideline called Koch’s postulate to
be used to establish cause of infectious disease.
Koch’s postulate states that
● Suspected pathogen must be present in all cases of disease and must not be present in
healthy individual
● The suspected pathogen must be isolated and grown invitro in a pure culture.
● Cells from pure culture of pathogen must cause disease in healthy individual when
introduced.
● The same pathogen must be re-isolated from infected organism.
Joseph Lister a British surgeon proposed that antiseptic surgical procedure reduces danger of
microbial infection after surgery, thus championed the use of carbolic acid as an antiseptic, such
that it became the first widely used antiseptic in surgery. He is acknowledged as “Father of
Antiseptic Surgery”.
Alexander Fleming a Scottish bacteriologist accidentally discovered the antibiotic in 1928,

when he came back from a vacation and found that a green mold called Pennicilium notatum
had contaminated Petri dishes in his lab and were killing some of the bacteria he'd been
growing. Penicillin was first called “mould juice.”
Ivanovsky & Beijerinck discovered infectious substances in tobacco plant and called it
tobacco mosaic virus (TMV). Thus the first virus ever discovered.
1.4 HISTORY OF THE DISCOVERY OF MICRO-

ORGANISM
▪ Theory of Spontaneous Generation (Abiogenesis)
Humans have been asking for millennia: Where does new life come from? Religion,
philosophy, and science have all wrestled with this question. One of the oldest
explanations was the theory of spontaneous generation, which can be traced back to
the ancient Greeks and was widely accepted through the Middle Ages.
It suggests that living creatures and organisms arises from nonliving matter or source
and that such occurrence is commonplace and regular. For instance, it was hypothesized
that certain forms of fleas and maggots could arise from inanimate matter such as dust
and decaying flesh respectively, mice simply appeared from grains stored in barns and
frogs arose from muddy banks of stale water.
This theory was logically synthesized by Aristotle who gathered and expanded on the
work of earlier philosophers.
The theory of spontaneous generation is in total contrast to the “Theory of
Equivocal Generation” and “Theory of Univocal Generation”.
While Equivocal Generation Theory suggests that living creatures and organisms arises
from unrelated living organisms such as tapeworms arising from the gut of man (Host),
Univocal Generation Theory on the other hand contrasts the former by suggesting that
living organisms arises from effective exclusive reproduction from genetically related
parent(s) of the same species such as man from man, this is known as Biogenesis.
▪ Conflict Arising from Spontaneous Generation

The idea of spontaneous generation brought about a great and concerned conflict
among scientist in the 17th and 18th century. While some supported it others debunked
and disproved its validity. Among the supporters were
John Needham (1748): Published a report of his experiment, in which he briefly boiled
a broth containing both plant, animal and microbial matter, hoping to kill all preexisting
microbe, he then sealed the flask. After few days he observed that the broth had
become cloudy and contained numerous microscopic creatures. He argued that the new
microbe must have arisen spontaneously. In reality however, he likely did not boil the
broth enough to kill all preexisting microbes because the boiling was brief.
Van Helmont: Proposed that mice could arise from rags and wheat kernels left in an
open container for weeks due to presence of mould.
Among the non-supporters were
Lazzaro Spallanzani (1751): Disagreed with Needham’s conclusion through carefully

executed experiment using heated broth. Both sealed and unsealed containers were
infused with animal and plant matter. He observed that heated but sealed containers
remained clear without any sign of spontaneous growth unless the flask was
subsequently opened to the air. This suggest that microbes were introduced into these
flasks from the air.
Francesco Redi (1668): Performed an experiment that was one of the first to refute
the idea that maggots spontaneously arise from meat left in open air. He predicted that
preventing flies access to meat would also prevent the appearance of maggots. He left
meat in six containers, two were open to air, two were covered with gauze, and two
were tightly sealed. His hypothesis was supported when maggots were observed in the
open container, but no maggot was observed in either the gauze covered container or
the tightly sealed container. He concluded that maggots could only develop when flies
were allowed to lay eggs on the meat, and that maggots were the offspring of flies not
the product of spontaneous generation.
ANALOGY OF REDI’S EXPERIMRNT (SOURCE: LUMENLEARNING.COM)

Louis Pasteur (1861): A prominent French chemist irrefutably disproved the theory of
spontaneous generation by an award winning intelligent experiment. He made a series
of flask with long twisted neck in which he boiled broth to sterilize it. His design allowed
air inside the flask to be exchanged with air outside, but prevented the introduction of
any airborne microorganisms, which would get caught by the twist and bends of the
flasks’ neck. He observed that sterilized broth in the twisted neck flask will remain
sterile as long as the twisted neck remained intact. However, should the neck be broken,
microbes would be introduced, contaminating the flask and allowing microbial growth in
the broth. He is credited for founding the assertion that “life only comes from life”. His
experiment earned him the prestigious Alhumbert Prize from the Paris Academy of
Sciences in 1862.
Never again was the theory of spontaneous generation argued over or believed.
He is considered the Father of modern microbiology.
ANALOGY OF PASTEUR’S EXPERIMRNT (SOURCE: LUMENLEARNING.COM)
1.5 GERM THEORY OF DISEASE

This is the currently accepted scientific theory for many diseases.
It states that “Many diseases are caused by the invasion, presence and actions of specific
microorganisms within the body”. It was proposed by Louis Pasteur.
Building on Pasteur’s work on germ theory, Robert Koch used experiment that proved that
Bacillus anthracis was the cause of anthrax. He extracted the bacterium from a sheep which
had died of anthrax, grew it invitro in a pure culture and injected a mouse with it. He observed
that the mouse developed the same disease as well. He went further to identify the bacterial
causes of tuberculosis and cholera.
Koch’s assistant Julius Richard Petri developed the Petri dish which makes the
observation of bacteria easier when grown in the laboratory.
SELF ASSESSMENT QUESTIONS

Multiple Choice
1. The study of diseases, its transmission, distribution and control over a given population
can be suitably referred to as
(a) Disease control (b) Public Health control (c) Public health and Epidemiology (d) All
of the above.
2. When the immune system and the impact of the activities of microorganisms on it is
being studied and investigated, this study can be suitably referred to as
(a) Physiology (b) Immunology and Immunochemistry (c) Anatomy (d)
Immunochemistry.
3. "The invasion of an organisms’ body tissue by pathogenic agents to multiply and cause
harm", this statement best fits which of the following terms. (a) Infection (b)
Disease (c) Invasion (d) Microbial Invasion.
4. The statement “life only comes from life” is accredited to which of the following
scientists (a) John Tyndall (b) Robert Koch (c) John Needham (d) Louis Pasteur.
5. With reference to the theory of spontaneous generation select the right pair.
(a) Spallanzani & Helmont (b) Pasteur & Redi (c) Needham & Pasteur (d) Needham &
Redi.
6. The use of series of flask with long twisted neck which prevented access of airborne
microorganisms into the flask to determine the authenticity of the theory of
spontaneous generation is accredited to (a) Louis Pasteur (b) Lazaro Spallanzani (c)
John Needham (d) Helmont.
7. _________ is considered the Father of modern microbiology. (a) Robert Koch (b)
Francesco Redi (c) John Lister (d) Louis Pasteur.
8. "Diseases are caused by the invasion, presence and actions of specific microorganisms
within the body”, this theory is called (a) Germ theory of diseases
(b) Theory of spontaneous generation (c) Theory of equivocal generation (d)
Theory of univocal generation.
9. According to Koch’s postulate, arrange the following in the right order
(i) Cells from pure culture of pathogen must cause disease in healthy individual when
introduced.
(ii) Suspected pathogen must be present in all cases of disease and must not be present
in healthy individual
(iii) The same pathogen must be re-isolated from infected organism.
(iv) The suspected pathogen must be isolated and grown invitro in a pure culture.
(a) ii, iii, i, iv (b) i, ii, iii, iv (c) ii, iv, I, iii (d) i, iv, iii, ii
10. The laboratory apparatus used for microbial growth and easy identification is______
and was developed by_______ (a) Conical flask, Edward Jenner (b)
Petri dish, Lillian Petri (c) Conical flask, Louis Pasteur (d) Petri dish,
Julius Petri.
11. Which of the following is incorrect
(a) Francesco Redi - Refuted the idea that maggots spontaneously arise from meat left
in open air. He predicted that preventing flies access to meat would also prevent the
appearance of maggots.
(b) Ferdinard Cohn - German biologist that discovered the heat resistant endospores of
bacteria.
(c) Antoni Van Leeuwenhoek - discovered red blood cells, spermatozoa cells and
infectious substances in tobacco plant and called it tobacco mosaic virus (TMV).
(d) Julius R. Petri - Developed the Petri dish for easy microbial growth and identification.
12. A chemical substance produced by the body to combat infectious agents as a result of
response to an antigenic stimulus is referred to as (a) Antigen (b) Antibody
(c) Immune (d) Blood
13. A Pandemic can be accurately defined as (a) The

outbreak of disease in a particular region or population (b) An epidemic of
infectious disease that has spread across a large region/continent
(c) A disease that exist permanently in a particular region or population (d) A
degenerative disease that is highly communicable and contagious and has the capacity
and potency to cause harm to a population of living things.
14. The Germ theory of diseases is accredited to (a) Louis Pasteur

(b) Ivanovsky (c) Antoni Van Leeuwenhoek (d) Robert Koch
15. Viruses were first discovered by (a) Louis Pasteur (b) Ivanovsky & Beijerinck
(c) Antoni Van Leeuwenhoek (d) Robert Koch
16. Sperm cells, red blood cells and animalcules were first discovered by (a) Louis Pasteur
(b) Ivanovsky & Beijerinck (c) Antoni Van Leeuwenhoek (d) Robert Koch
17. Which of the following disease is considered a Pandemic (a) Malaria (b) Covid - 19
(c) Dysentery (d) Measles.
18. A substance of chemical complexity that kills bacteria out-rightly is best called (a)
Bactericidal (b) Bacteriostatic (c) Bacterioinhibiting (d) None of the above.
19. Which of the following is incorrect (a)Epidemic – The outbreak of disease in a

particular region or population over a certain period of time (b)Endemic – A
disease that exist permanently in a particular region or population
(c)Pandemic – An epidemic of infectious disease that has spread across a large
region/continent (d) None of the above
20. A process for sterilizing substances usually food substances that kills heat resistant
endospores is called (a) Sterilization (b) Tyndallization (c) Disinfection
(d) Pasteurization
21. Which of the following is incorrect (a) Theory of spontaneous generation - It suggests
that some living creatures and organisms arises from nonliving matter or source
(b) Theory of Univocal generation - suggests that living organisms arises from
reproduction from genetically related parent(s) of the same species (c) Equivocal
Generation Theory - suggests that living creatures and organisms arises from unrelated
living organisms (d) Germ theory of disease - many diseases
are caused by the invasion, presence and actions of specific microorganisms within the
body
22. Serum can be defined as (a) A clear yellowish fluid obtained upon
separating coagulated blood into its solid and liquid components. (b) A clear
yellowish fluid obtained upon separating uncoagulated blood into its solid and liquid
components. (c) A clear yellowish fluid obtained upon separating dried coagulated
blood into its solid and liquid components. (d) A clear yellowish
fluid obtained upon separating dried blood into its solid and liquid components.
23. _________ proved that Bacillus anthracis was the cause of anthrax. (a) Robert Koch
(b) John Tyndall (c) Joseph Lister (d) Ferdinard Cohn
24. Antiseptic surgical procedures that reduces the chances of wound infection can be
credited to (a) Robert Koch (b) John Tyndall (c) Joseph Lister (d)
Ferdinard Cohn
25. In 1876, heat resistant form of bacteria was discovered by (a) Robert
Koch (b) John Tyndall (c) Joseph Lister (d) Ferdinard Cohn
26. Microbiology as a science started with (a) Louis Pasteur (b) Ivanovsky & Beijerinck (c)
Antoni Van Leeuwenhoek (d) Robert Koch
27. "Worms found on rotten meat originates from egg of flies and not from Spontaneous
Generation " this statement is credited to (a) Louis Pasteur (b) Lazaro
Spallanzani (c) Francesco Redi (d) Helmont.
28. Agar can be defined as (a) A solidifying agent usually introduced to

solidify liquid nutrient media in the laboratory. (b) A solidifying agent
usually introduced during food production in manufacturing companies. (c) A
solidifying agent usually introduced when population of microbial life in liquid nutrient
media exceeds the optimum. (d) A chemical agent usually used in the
laboratory.
29. Culture can be defined as (a)A collection or population of microbial growth in
a solid nutrient media. (b) A collection or population of
microbial growth in a liquid nutrient media.
(c) A collection or population of microbial growth in a nutrient media. (d) A way of life of
a people.
30. A biological substance constituted of weakened or killed form of microbes, its toxins or
its surface protein, that when introduced into a living host, stimulates production of
antibodies and induces immunity against one or several diseases (a) Antigen (b)
Serum (c) Plasma (d) Vaccine
31. Agricultural products both harvested and unharvested were observed to be attacked by
unseen agents resulting in rot and decay of these products. The most suitable
professional to be invited to diagnose the situation is a (a) Veterinary doctor (b)
Agricultural Microbiologist (c) Farm Manager (d) A microbiologist.
32. Agar a solidifying agent is sourced from (a) Algae (b) Rhodohytes (c)
Bryophytes (d) Tallophytes
33. In 1962, in the midst of an epidemic, individuals were quickly administered with
vaccines by certified health officials, this activity was solely aimed at
(a) Stimulating antigen production in the body (b)
Administering a cure to such disease. (c) Stimulating a
sustainable healing process. (d) Stimulating antibody
production in the body
34. A patient was observed to display rare and unusual signs and symptoms by a certified
health official. In order to diagnose the nature of such disease, it is expedient that the
services of a (a) Microbiologist be consulted (b) Epidemiologist be consulted
(c) Medical Microbiologist be consulted (d) Immunologist be
consulted
35. The scientist that finally laid to rest the theory of spontaneous generation was (a)
Robert Koch (b) John Tyndall (c) Joseph Lister (d) Louis Pasteur
36. The scientific study of algae is called (a) Algeology (b) Phycology (c) Mycology
(d) None of the above
37. The organic matter used by Francesco Redi in his experiment was a (a) Meat
(b) Pork meat (c) Fish (d) Plant product.
38. Identify the correct pair (a) Sterilization and Antiseptic (b)
Bacteriostatic and Sterilization (c) Sterilant and Disinfectant (d) Sterilization
and Bactericidal
39. Identity the correct pair (a) Robert Koch & Ivanovsky (b) John
Tyndall & Spallanzani (c) Louis Pasteur & Francesco Redi (d) Joseph Lister &
Louis Pasteur
40. The study and diagnosis of disease through examination of organs, tissues, bodily fluids,
and whole bodies is called (a) Epidemiology (b)
Pathology (c) Pathogenicity (d)Pathogenesis.
41. The discovery of penicillin is credited to (a) Robert Koch (b) John Tyndall
(c) Alexander Fleming (d) Joseph Lister
42. The microbial source of penicillin is (a)Pennicilium notatum (b)Pennicilium
pallidum (c) Treponema pallidum (d) Mycobacterium
leprae

Theory
1. Succinctly explain the theory of spontaneous generation and why people once accepted
it as an explanation for the existence of certain types of organisms
2. Explain the conflict that arose between scientists over the theory of spontaneous
generation
3. Amaka is a 19-year-old college student living in the dormitory. In January, she came
down with a sore throat, headache, mild fever, chills, and a violent but unproductive
(i.e., no mucus) cough which became highly contagious. To treat these symptoms,
Amaka began taking an over-the-counter cold medication, which did not seem to work.
In fact, over the next few days, while some of Amaka’s symptoms began to resolve,
however, her cough and fever persisted coupled with a congested respiratory tract, and
she felt very tired and weak. (a) Predict the most likely respiratory
disease Amaka is currently facing. (b) Predict the most likely causal organism of this
infectious disease. (c) Is it an endemic, pandemic or epidemic disease? Defend
your answer (d) Predict the most likely way Amaka got infected with
this disease. (e) Using Koch postulate, systematically enumerate the steps to
take to establish the cause of this infectious disease. (f) Enumerate 3
preventive measures to be taken against this disease
4. Tony a dedicated bricklayer lives in a water logged area in a Nigerian state. One day he
returned from work with severe symptoms of fatigue and head ache. After dinner he
self medicated and expected to be fine shortly. But he observed that after 6hours there
was no relief instead additional symptoms of joint pain, fever and bitter taste in mouth.
(a) Using this symptoms predict a name for Tony's ailment (b) Predict the
most likely way Tony got infected with this disease. (c) Is it an endemic,
pandemic or epidemic disease (d) Predict the most likely causal
organism of this disease. (e) Is it a communicable disease? Defend your
answer (f) Enumerate 3 preventive measures to be taken against this
disease.
5. Differentiate between the Theory of equivocal generation and Theory of univocal

generation with relevant examples.
6. Enumerate the contribution of the following scientists to microbiology. (i) Louis Pasteur
(ii) Antoni Van Leeuwenhoek (iii) Robert Koch (iv) John Tyndall (v) Lazaro
Spallanzani
7. Differentiate between Shelf life, Best before date and Expiring date.
8. Exposure to air is necessary for microbial invasion and growth. TRUE or FALSE. Justify
your answer
9. Theory of spontaneous generation suggests that some living creatures and organisms
arises from nonliving matter or source. TRUE or FALSE. Justify your answer.
10. All diseases are caused by the invasion of microorganisms into a host. TRUE or FALSE.
Justify your answer.
11. All infections will eventually lead to disease. TRUE or FALSE. Justify your answer.
ANSWERS (Multiple Choice)

1. C
2. B
3. A
4. D
5. B
6. A
7. D
8. A
9. C
10. D
11. C
12. B
13. B
14. A
15. B
16. C
17. B
18. A
19. D
20. B
21. A
22. A
23. A
24. C
25. D
26. C
27. C
28. A
29. C
30. D
31. B
32. B
33. D
34. C
35. D
36. B
37. A
38. D
39. C
40. B
41. A
42. C
THEORY
1. Make reference to chapter 1.4
3. (a) Covid - 19 (b)Corona Virus (c) Pandemic. It's a pandemic disease because it
has spread across many regions and continent during a particular time frame.
(d) Inhalation of aerosols from infected person.
(e)
● Suspected pathogen must be present in all cases of disease and must not be present in
healthy individual
● The suspected pathogen must be isolated and grown invitro in a pure culture.
● Cells from pure culture of pathogen must cause disease in healthy individual when
introduced.
● The same pathogen must be re-isolated from infected organism
(f)
● The use of effective nose covering or mask when in public space.
● Maintaining always effective social distance of about 2 metres when in public space.
● The practice and sustainance of personal hygiene and frequent hand washing.
4. (a) Malaria (b) A bite by an infected female anopheles mosquito. (c)

Endemic (d) Plasmodium malariae (e) No. It is
not communicable because it cannot be transferred or passed from one person to
another. (f)
● The practice and sustainance of personal hygiene and frequent sanitation of our
immediately environment.
● The use of treated and recommended mosquito net when asleep.
● Consistent fumigation of immediate environment using an insecticide.
8. True, because air contains a wide range of variety of impurities which includes
microorganisms.
9. False, because the theory of spontaneous generation suggests that ALL and not SOME
living creatures and organisms arises from nonliving matter or source.
10. False, because some diseases are not caused by the invasion of microorganisms e.g.
Diabetes, hypertension etc
11. False, not all infection eventually lead to disease as the host may have strong
immunity and not susceptible to the invading pathogen.
CHAPTER 2
2.1 TYPES AND TAXANOMIC GROUPINGS OF

MICROORGANISMS
Microorganisms can be suitably classified into 2 major groups namely: Prokaryote &
Eukaryotes.
Prokaryotes are living unicellular microscopic entities with a cell that lacks a well defined
nucleus, and have a simplified internal organization, thus their nuclear material is termed
“Nucleoid”. Example includes Bacteria and Archaea.
Eukaryotes are living entities usually multicellular and macroscopic while few are unicellular
and microscopic with a cell that has a well-defined nucleus, membrane bound organelles, and
complex internal organization. Example includes Fungi, Algae, Protozoans, Plant and Animal.
NB: Viruses are not considered to be in this classification because they are acellular and thus
are considered to be particles.
2.2 DIFFERENCES BETWEEN PROKARYOTES & EUKARYOTES

PROKARYOTIC CELL EUKARYOTIC CELL
It lacks a membrane bound nucleus, thus It has a well-defined membrane bound

possesses a nucleoid. nucleus.
It lacks the presence of cellular organelles. Cellular organelles are abundantly present.
The DNA is single and circular in shape and The DNA is many and linear in shape and
appearance. appearance.
It possess 70s ribosome. It possess 80s ribosome.
They are unicellular organisms. They are mostly multicellular but few are
unicellular.
Ribosomes are located on the plasma Ribosomes are located on or within cellular
membrane and not on or within any cellular organelles such as rough endoplasmic
organelle. reticulum, mitochondria, nucleus etc.
In photosynthetic cells, chlorophll is located In photosynthetic cells, chlorophyll is located

at the plasma membrane. at the chloroplast in the cytoplasm of the cell.
They reproduce only asexually through binary The reproduce asexually through spores,
fission. fragmentation etc. and sexually through
conjugation.
Their chromosomes are restricted to the Their chromosomes is restricted to the

nucleoid. nuclear membrane.
SIMILARITIES BETWEEN PROKARYOTES & EUKARYOTES.
● They both possesses genetic materials.
● They both have a cytoplasm.
● They both have a plasma or cell membrane.
● They both possesses ribosomes.
● They both possesses enzymes.

● They could both be parasitic though some are non-parasitic.
2.3 VIRUSES
A virus is an obligate intracellular parasite with genome of either RNA or DNA which is enclosed
in a particular protein coat called capsid that cannot be seen by light microscope but only by an
electron microscope. Viruses are obligate intracellular meaning they can only replicate and
survive inside a living host.
STUCTURE OF AN ENVELOPED VIRUS (SOURCE: BIOLOGY DICTIONARY.NET)
Viruses are acellular, meaning they are biological entities that do not have a cellular structure.
Therefore, they lack most of the components of cells, such as organelles, ribosomes, and the
plasma membrane. They much more smaller than bacteria with a diameter of 20 -
300nanometers.
A virus in its extracellular state is called a Virion. Viruses are considered non-living things
because they lack the ability to survive outside a living host.
A Viroid is the smallest infectious RNA molecule that has no capsid. They are much
smaller than viruses, and like viruses they can only replicate in a living cell.
A Prion is a proteinaceous infectious particle that is responsible for at least 7 neuro-

degenerative diseases.
Viruses, viroids, and prions are non-living things hence are termed agents or
particles.
HISTORY OF VIRUSES
The scientific study of viruses began in 19th century. Louis Pasteur and Edward Jenner
developed the first vaccine against viral infection although they did not know that viruses
existed. In 1892, Dmitry Ivanovsky used a filter that had pores small enough to retain bacteria
to show that sap from a diseased tobacco plant remained infectious to healthy tobacco plant
despite having been filtered.
Martinus Beijerinck called the filtered infectious substance a “virus” and
this is considered to be the beginning of virology.
Thomas Milton Rivers was the first to define virus as obligate parasite which means that viruses
cannot replicate and survive outside of a living host.
Wendell Stanley of University of California demonstrated that viruses are particles rather than a
fluid.
TERMS IN VIROLOGY
● Capsid: the outer protein shell of a virus.
● Envelope: an enclosing structure or cover, such as a membrane.
● Filamentous: to have the form of threads or filaments.
● Isometric: of, or being a geometric system of three equal axes lying at right angles to
each other (especially in crystallography)
● Capsomere: Any of the individual protein subunits of a viral capsid
● Icosahedral: of, relating to, or having the shape of an icosahedron
2.4 VIRAL CLASSIFICATION

Viruses can be classified as
● Enveloped or Non-enveloped Viruses.
● DNA or RNA viruses.
Enveloped viruses: These are viruses that has an outer wrapping or covering called envelope.
Thus envelope protects the genetic material in their life-cycle when traveling between host
cells. The envelopes are typically derived from portions of the host cell membranes, but include
some viral glycoproteins. (See diagram in 2.3)
Example of Enveloped viruses includes: Poxvirus (DNA), Herpesvirus (DNA), Hepadnavirus
(DNA), Flavivirus (RNA), Coronavirus (RNA), Rhabdovirus (RNA), Filovirus (RNA), Hepatitis D.
Non-enveloped Viruses: These are viruses that do not have viral envelopes covering their
central Capsid but are composed mainly of capsid protein and nucleic acid (DNA or RNA), vis-a-
vis nucleocapsid., which constitute an infectious unit. Whereas enveloped viruses are
composed of an envelope and nucleocapsid.
Non-enveloped viruses can be more resistant to changes in temperature, pH, and some
disinfectants than are enveloped viruses.
Example of non enveloped viruses includes: Papillomaviridae (DNA), Adenoviridae (DNA),
Picornaviridae (RNA), Caliciviridae (RNA).
2.5 SHAPES OF VIRUS

Helical – These viruses are composed of a single type of capsomer stacked around a central axis
to form a helical structure, which may have a central cavity, or hollow tube.
Icosahedral – Most animal viruses are icosahedral or near-spherical with icosahedral symmetry.
Prolate – This is an isosahedron elongated along one axis and is a common arrangement of the
heads of bacteriophages.
Envelope – Some species of virus envelop themselves in a modified form of one of the cell
membranes, either the outer membrane surrounding an infected host cell or internal
membranes such as nuclear membrane or endoplasmic reticulum, thus gaining an outer lipid
bilayer known as a viral envelope.
Complex – These viruses possess a capsid that is neither purely helical nor purely icosahedral,
and that may possess extra structures such as protein tails or a complex outer wall. Some
complex viruses are large enough to be visible with a light microscope.
2.6 BACTERIOPHAGE
Bacteriophage is a virus with double stranded DNA genome that infects and replicates within
bacteria and archaea, it was discovered and characterized by Frederick Twort and Felix
d’Herelle.
STRUCTURE OF A BACTERIOPHAGE (SOURCE: RESEARCHGATE.NET)
2.7 REPLICATION CYCLE OF VIRUSES IN HOST CELL.
REPLICATION CYCLE OF A VIRUS IN A HOST CELL (SOURCE: SPARKNOTE LLC)
All viruses share 6 basic steps in their replication cycles. These are;
1. Attachment 2. Penetration 3. Uncoating 4. Replication 5. Assembly 6.Release.
As shown above , the virus must first attach itself to the host cell. This is usually accomplished
through special glycoprotiens on the exterior of the capsid, envelope or tail.
Next, penetration occurs, either of the whole virus or just the contents of the capsid. If the
entire capsid enters, the genetic material must be uncoated to make it available to the cell's
replication machinery. Replication of genetic material takes place, as well as the production of
capsid and tail proteins. Once all of the necessary parts have been replicated, individual virus
particles are assembled and released. Release often takes place in a destructive manner,
bursting and killing the host cell.
Some viruses have a slightly more complicated replication cycle involving lytic and lysogenic
phases.
The lytic phase is similar to that described above, with virus particles infecting and being
replicated. In the lysogenic phase, however, viral genetic material that has entered the host cell
becomes incorportated in the cell and lies dormant. It is passed on to the progeny of the
infected cells. Eventually, the lytic phase will start again, and cells that were never infected
themselves, but carry the viral genetic material will begin to produce new virus particles.
2.8 FUNCTIONS OF THE STRUCTURES OF A VIRUS

ENVELOPE PROTEIN:
● Serves to identify and bind viruses to receptor sites on the host’s membrane.
VIRAL ENVELOPE:
● It consists of protein and phospholipid membranes derived from the host cell.
● Confers external protection to the viral particle against predation and harsh
environmental conditions.
● Helps viruses avoid the host immune system.
CAPSID:
● It determines the shape of a virus.
● it protects the nucleic acid from digestion by enzymes.
● Contains special sites on its surface that allows the virion to attach to a host cell.
● Provides proteins that enable the virion to penetrate the host cell membrane.
GENOME:
● Carries the genetic information if the viral particle.
● It exist as either DNA or RNA, Single Stranded or Double Stranded, Circular or Linear in
shape.
ANALYSIS OF VIRAL GENOME. (SOURCE: SCIENCE DIRECT)

2.9 Characteristics of Viruses
● They are obligate intracellular parasite, thus cannot survive outside a living host.
● They can infect both plant and animal cell even other microorganisms.
● Viruses that infect bacteria are called bacteriophages and those that infect fungi are
called mycophages and some that infect other viruses are called virophages.
● They are acellular, i.e. they contain no cytoplasm and cellular organelles.
● They are capable of mutation.
● They carry out no metabolism of their own and uses the host cell metabolic machinery.
● They cannot be grown in synthetic culture media as they lack metabolic machinery of
their own, but can be grown in embryonated eggs or cell cultures.
● They cannot survive when exposed to high temperature and can be crystallized under
very low temperature
● The combination of the viral capsid and nuclear material is known as Nucleocapsid.
● Their nuclear material is made up of either RNA or DNA but not both at same time.
● Their nuclear material is either double stranded or single stranded but not both at the
same time
● They can exist as enveloped i.e. having an envelope covering the nucleocapsid or non-
enveloped i.e. no envelope over the nucleocapsid.
● They exist in different shapes i.e. Isometric, Icosahedral, and Helical symmetry.
● The shape of a virus is determined by the shape of the capsid.
● Capsids are composed of many identical protein subunits called capsomeres.
● All bacteria and animal virus but not plant virus are able to attach to a specific receptor
site on a host cell through a process called Adsorption.
2.10 VIRAL DISEASES, CAUSATIVE AGENTS AND MODE OF INFECTION

VIRAL DISEASES CAUSATIVE AGENTS MODE OF INFECTION
Small pox Variola Virus ● Person to person

spread via infected
aerosols and air
droplets.
● Transmission may also
occur via
contaminated
clothing and bedding
(less common).
Yellow fever Flavi Virus ● Infected mosquitoes

of the Aedes aegypti
specie transmit the
virus from person to
person.
Rabbies Rhabdo Virus ● Virus is usually

transmitted to
humans by infected
saliva through the bite
of a rabid animal such
as dogs.
Ebola Virus Disease (EVD) Ebola Virus ● Ingestion of raw or

not fully cooked and
contaminated meats.
● However, human-to-
human transmission is
via close contact
which is the major
route of current
outbreaks.
Acquired Immuno-Deficiency Human Immunodeficiency ● Sharing of
Syndrome (AIDS). Virus contaminated
injection drug syringe
or needle.
● Sexual intercourse
with infected person.
● Transmission from
infected mother to
child.
● Exposure to infected
blood or blood
products.
● And transplantation
of infected organs or
tissues to healthy
individual.
Chicken pox Varicella zoster Virus ● Mostly airborne,

spread by coughing
and sneezing.
● Direct contact with
skin lesions.
Hepatitis B Hepatitis B Virus (HBV) ● Perinatal transmission

is the main route of
infection.
● Whereas in areas with
low HBV endemicity,
sexual contact
amongst high-risk
adults is the
predominant route.
Measles Measles Virus ● Direct contact from

person-to-person.
● Airborne spread as
infectious droplets
from the respiratory
secretions of a patient
with measles can
remain airborne for
up to two hours.
Polio Polio Virus ● Person-to-person

spread via the fecal-
oral route
Covid - 19 Corona Virus ● Predominantly

transmitted through
contact with infected
respiratory droplets
either as aerosol or as
fomites.

Multiple Choice
1. Viruses are said to be acellular because (a) They lack cellular structures (b) They
lack cellular metabolism (c) They have cellular structures (d) They have
cellular metabolism
2. The extracellular inert form of a virus is called (a) Prion (b) Virus (c) Virion
(d) Viroid
3. The combination of the viral capsid and nuclear material is known as (a)
Nuclearcapsid (b) Nucleocapsid (c) Nucleicapsid (d) None of the above
4. The smallest known infectious agent in existence is (a) Bacteria (b) Algae (c)
Fungi (d) Virus
5. The first step in virus replication cycle is (a) Penetration (b) Elongation (c)
Attachment (d) Uncoating
6. An infectious disease known to man that can be transmitted from person to person via
an infected Aedes aegypti mosquito is (a) Lassa Fever (b) Yellow fever (c)
Malaria (d) Cholera
7. Ade on his way back from school was accosted by a mad wild dog. He tried to escape
but was chased by the dog who left a wound on his leg after a successful bite. What
disease is he likely to contract? (a) Tissue Inflammation (b)
Rhabies (c) Chronic fever (d) Lassa Fever
8. The shape of a virus is determined by the (a) Capsid (b) Viral Envelope (c)
Capsomere (d) Viral Genome
9. The second step in virus replication cycle is (a) Penetration (b) Elongation (c)
10. The following except one are able to attach to a specific receptor site on a host cell (a)
Bacteria (b) Bacteria virus (c) Animal virus (d) Plant virus
11. The attachment of a virus to a specific receptor site on a host cell is through a process
called (a) Adhesion (b) Absorption (c)Adsorption (d) Attachment
12. Viruses exist in different shapes except
(a)Icosahedral (b) Isometric (c) Isohelical (d) Helical
13. The smallest infectious RNA molecule that has no capsid .(a) Prion (b) Virus
(c) Viroid (d) Virion
14. Viruses are best grown invitro in a laboratory using (a) Cell culture (b) Nutrient media
(c) Synthetic media (d) Media culture.
15. A viral structure that provides proteins that enable the virion to penetrate the host cell
membrane is called (a)Genome (b) Envelope (c) Tail (d) Capsid
16. The major distinguishing factor between eukaryotic and prokaryotic cell is in the
presence of (a) A well defined nucleus (b) Cellular organelles (c) 80s
ribosome (d) linear shaped DNA.
17. The third step in virus replication cycle is (a) Penetration (b) Release (c) Attachment
(d) Uncoating
18. Viruses are best considered to be (a) Eukaryotes (b) Monera (c) Prokaryotes
(d) Particles
19. Which of the following does not support the replication and existence of a virus (a)
Aquatic environment (b) High temperature environment (c) Low temperature
environment (d) Humid environment
20. Which of the following structures confers external protection to the viral particle against
predation and harsh environmental conditions. (a) Envelope (b) Genome
(c) Envelope protein (d) Capsid
21. From the physiology and metabolism of viruses it is evident that viruses are best fit to
be called (a) Autotrophs (b) Heterotrophs (c)
Chemoautotrophs (d) None of the above
22. These viruses possess a capsid that is neither purely helical nor purely icosahedral, and
that may possess extra structures such as protein tails (a) Helical viruses (b) Simple
viruses (c) Complex viruses (d) Icosahedral viruses
23. A bacteriophage is a virus with (a) Double stranded DNA genome (b) Double
stranded RNA genome (c) Single stranded DNA genome (d) Single stranded RNA
genome by
24. Which of the following supports the crystallisation of a virus (a) Aquatic environment
(b) High temperature environment (c) Low temperature environment (d) Humid
environment
25. Which among all viral structure carries the genetic information of the viral particle. (a)
Nucleus (b) Nucleoid (c) Genome (d) Chromosome
26. __________ viruses can be more resistant to changes in temperature, pH, and some
disinfectants. (a) Simple viruses (b) Non-enveloped viruses (c) Enveloped viruses
(d) Complex viruses
27. The last step in virus replication cycle is (a) Penetration (b) Release (c)
28. _________ developed the first vaccine against viral infection (a) Louis Pasteur (b)
Edward Jenner & Robert Koch (c) Louis Pasteur and Joseph Lister (d) Louis Pasteur &
Edward Jenner
29. The first ever discovered virus was from a (a) Tobacco plant (b) Cocoa plant
(c) Tobacco seed (d) Cocoa seed
30. The head and tail of a bacteriophage is joined together by a structure called (a)
Connecting fibre (b)Fibre (c) Sheath (d)Collar
31. One of the following is a good example of an enveloped virus with an RNA genome
(a)Pox virus (b)Herpes virus (c) Corona Virus (d) Adenoviridae
32. The definition of virus as an obligate parasite which means that viruses cannot replicate
and survive outside of a living host can be credited to (a) Ivanovsky (b) Martinus
Beijerinck (c) Thomas Rivers (d) Wendell Stanley
33. Viruses are considered non-living things because (a) they lack the
ability to survive outside a living host. (b) they lack the ability to replicate
(c) they lack the ability to reproduce (d) All of the above
34. The acronym AIDS stands for (a) Acquired Immuno-Deficiency Syndrome (b)
Acquired Immuney Deficiency Syndrome (c) Acquired Immuno-Deficiency Sickness (d)
Acquired Immuney Deficiency Sickness.
35. The causal organism of yellow fever is (a) Variola virus (b)Varicella zoster Virus (c)
Adenoviridae (d) Flavivirus
36. A proteinaceous infectious particle implicated in various neuro-degenerative diseases is
(a) Virus (b) Prion (c) Viroid (d) Virion
37. When a viral particle is observed using a light microscope which of the following
structures will be observed (a) Genome (b) Capsid (c) Envelope (d) None of the above
38. When a viral particle is observed using an electron microscope which of the following
structures will be observed (a) Genome (b) Capsid (c) Envelope (d) All of the above
39. Viruses that infect other viruses are called (a) Viriophage (b) Mycophage
(c)Parasite (d) Bacteriophage
40. Tolu, a cleaner at a health centre is tasked with eliminating viruses from the surfaces of
inert objects. What suitable chemical substance should be used (a) Antiseptic Substance
(b) Viricidal Substances (c) Disinfectant (d) Bleach.

Theory
1. Differentiate between the following (i) Viroid (ii) Virus (iii) Virion (iv) Prion
2. "A virus is an obligate intracellular parasite that is considered as a particle." Explain

each of the underlined term in the statement above.
3. On a windy day, Ade who is a popular healthy road traveller had a tragic road accident
around a rural environment and he required urgent blood transfusion. Due to absence
of quality medical care availability in such areas, he was unfortunately rushed to a
substandard medical facility filled with under trained medical personels. Unscreened
blood was transfused into his system with the aid of unsterilized needles. Fortunately,
Ade survived but after couple of months, he observed exponential weight loss, loss of
appetite, chronic cough with blood stained sputum, general body weakness, diarrhoea,
chronic fever, joint aches, headache etc. (a) Using this
symptoms predict a name for Ade's ailment. (b) Predict the most likely
causal organism of this infectious disease. (c) Is it an endemic, pandemic or
epidemic disease? Defend your answer (d) Predict the most likely way
Ade got infected with this disease (e) Enumerate 3 preventive
measures to be taken against this disease
4. With the aid of a diagram, show and explain two function each for the following viral
structures (i) Genome (ii) Envelope (iii) Envelope protein (iv) Capsid
5. Comprehensively explain the basic steps of replication of a virus in a host cell.
6. State 5 differences between Enveloped and Non-enveloped Viruses
7. Write a brief essay on the various shapes of viruses.
8. Explain any 5 viral disease under the following heading (i) Name of causal organism
(ii) Mode of transmission (iii) Preventive measures
9. Viruses are the smallest infectious particle on earth. TRUE or FALSE (b) Justify
your answer
10. State 5 Economic importance of viruses
11. Viruses may not be Prokaryotes or Eukaryotes but are living things. TRUE or FALSE
(b) Justify your answer

1. A
2. C
3. B
4. D
5. C
6. B
7. B
8. A
9. A
10. D
11. C
12. C
13. C
14. A
15. B
16. A
17. D
18. D
19. D
20. A
21. D
22. C
23. A
24. C
25. C
26. B
27. B
28. D
29. A
30. D
31. C
32. C
33. A
34. A
35. D
36. B
37. D
38. D
39. A
40. B
THEORY
1. Make reference to Chapter 2.3
2. Obligate indicates the vulnerability of a virus outside a living host thus viruses cannot
survive outside a host.
Intracellular indicates the virus survives by invading living cells of its host.
Parasite indicates the harmful ability and role viruses play when inside a host.
Particle indicates that viruses are considered non-living entities because they cannot
survive upon isolation from a host.
3. Acquired Immuno-Deficiency Syndrome.
Human Immunodeficiency Virus.
Pandemic, It's a pandemic disease because it has spread across many regions and
continent during a particular time frame.
Through the unscreened blood and/or the unsterilised needle.
● Sterilization of needles, clippers and other sharp object before use
● Effective testing and screening of blood sample before tranfusing into a patient.
● Abstinence from engaging in multiple sex partner and pre-marital sex.
4. Make reference to Chapter 2.3 & 2.8

9. True. They are the smallest infectious particle on earth because their diameter
measures 20 - 300 nanometers, the smallest diameter of microbial entity ever known
unlike bacteria which is 1 - 10 micrometer.
10. Viruses are implicated in several human, plant and animal diseases
Viral attenuated parts or its protein are modified and purified to produce vaccine which
confers active immunity on the recipient.
Viruses such as bacteriophage are employed in the infection and termination of
targeted bacteria cells especially in water purification
11. FALSE. Viruses are considered non-living things because they lack the ability to survive
outside a living host.
CHAPTER 3
3.1 BACTERIA
These are unicellular microscopic entities which possesses a peptidoglycan layer within it's cell
wall, a genetic material called nucleoid, and no cellular organelles thus are regarded as
Prokaryotes . They constitute a large domain of prokaryotic microorganisms. Bacteria have a
number of shapes, ranging from spheres to rods and spirals.
They are relatively small (1 – 10 µm) and simple, single-celled organisms that has existed on
earth for over 3.5 billion years. These prokaryotes are most-likely the greatest distribution of all
organisms.
They are ubiquitous meaning they are found everywhere from the soil to the air and even
water. In these various environments, bacteria can even withstand extreme temperatures and
chemically harsh environments.
Bacteria have developed numerous modes of nutrition. They can generally be classified as
either an autotroph or a heterotroph. These two nutritional modes can be further
subcategorized based upon whether they use light, inorganic or organic compounds. Most
prokaryotes, however, are heterotrophic, where they obtain nutrients from their surrounding
environment.
Bacteria were first observed by Antonie Van Leeuwenhoek in the year 1676, and he called them
'animalcules' (from Latin 'animalculum' meaning tiny animal). Allegedly, September 17, 1676
was the exact day when he reported the existence of bacteria.
Using single-lensed microscopes of his own design, he was the first to experiment with
microbes. Through his experiments, he was the first to relatively determine their size. Most of
the animalcules are now referred to as unicellular organisms, although he observed
multicellular organisms in pond water.
He was also the first to document microscopic observations of muscle fibers, bacteria,
spermatozoa, red blood cells, crystals in gouty tophi, and blood flow in capillaries.
STRUCTURE OF A BACTERIA CELL (SOURCE:

RESEARCHGATE.NET)
3.2 FUNCTIONS OF THE STRUCTURES OF A BACTERIAL CELL.

In the composition of a bacterial cell, there are numerous structures with different functions
which are arranged in this order;
FLAGELLA: A long thick whip-like structure composed of microtubules. It's made up of protein
flagellin and the filament ends with a capping protein. It's usually 20 nanometers thick.
Functions:
● It is useful in the locomotion of the cell.
● They propel cells in a whip-like motion.
PILI/FIMBRIAE: Hair-like appendages found on the surface of many bacteria and archea but not
all. They are made of protein and originate from cytoplasmic membrane. Pili are longer and
fewer in number than fimbriae. Functions:
● Enable cell adhere to a surface.
● Some generate motile forces e.g. type IV pilli (T4P).
● Sex pilli is involved in DNA transfer.
CAPSULE: A thick outermost structure that surrounds a bacterial cell, mostly present in Gram
negative bacteria and absent in Gram positive bacteria.
Functions:
● Protects the cell against predators and environmental assault..
● It assists the bacterial cell to twart the host defense immune system thus increases it's
virulence and pathogenicity.
● It aids attachment of bacterial cell to a surface.
CELL WALL: This is a rigid barrier that surrounds a bacterial cell and it's the second structure of
a gram negative bacteria but first structure of a gram positive bacteria. A thick layer of
peptidoglycan characterises the cell wall of gram positive bacteria.
Functions:
● Acts as a rigid barrier that prevents cytoplasmic contents from bursting out due to
osmotic pressure.
● It accommodates the peptidoglycan which provides rigidity to bacterial cell wall and
preventing the cell from lysing.
● Maintains shape and integrity of the cell.
● Acts as coarse filter for materials passing in and out of the cell.
PEPTIDOGLYCAN: This is the major structural component of a bacterial cell wall. It's a complex
molecule composed of alternating units of N-acetylmuramic acid (NAM) and N-
acetylglucosamine (NAG) cross-linked by short peptides, the result is a flat crosshatch pattern
that is strong and rigid yet open enough for movement of particles. It's substantially thicker in
Gram positive bacteria (20 - 80 nanometers) than in Gram negative bacteria (7 - 8 nanometers).
Functions:
● Confers rigidity on the cell wall preventing lysis of cell due to osmotic pressure.
● It also helps to counteract the osmotic pressure of the cytoplasm.
● Facilitates movement of particles in and out of the cell.
CYTOPLASMIC/CELL/PLASMA MEMBRANE: A phospholipid bilayer embedded with proteins

characterised by a hydrophilic head and hydrophobic tail.
Functions:
● Acts as a semi-permeable membrane that regulates passage of substances in and out of
the cell.
● Contains respiratory enzymes necessary for respiration.
● Contains pigments necessary for photosynthesis.
● Contains receptor proteins necessary to maintain cell integrity.
● Aids in ATP transfer.
● Contains antigen which is useful in pathogenicity.
CYTOPLASM: This is the internal cavity of bacterial cell comprises of cytosol (gel-like substance
enclosed within the cell membrane), cytoplasmic membrane and the genetic material nucleoid.
Functions:
● It's the site of metabolic activities e.g Glycolysis, cell division etc.
● Contains enzymes responsible for metabolism and the break down of waste.
● Also responsible for shape of the cell.
● It stores molecule necessary for cellular processes.
NUCLEOID: An irregularly shaped region within a bacterial cell, usually compacted to fit because
the length of the DNA chromosome is larger compared to the dimension of the cell.
Functions:
● Contains all or most of the genetic material of the cell.
● It includes enzymes and proteins that transcribe RNA & DNA.
● Aids in cell development and growth.
PLASMID: A small circular extra-chromosomal DNA structure capable of autonomous

replication. This extrachromosomal DNA is found in plasmids, which are small, circular, double-
stranded DNA molecules. Cells that have plasmids often have hundreds of them within a single
cell.
Functions:
● Carries genetic info that confers advantage to cell in certain conditions such as antibiotic
resistance.
● Facilitates replication of bacteria.
Other cellular content includes:

Ribosome (involved in protein synthesis). Gas vesciles (provides bouyancy to cell). Slime
(enables bacteria adhere to a surface).
3.3 SHAPES OF BACTERIA

Most common bacteria exist in two major shapes; spherical called coccus and cylindrical called
rod. A rod shaped bacteria is called Baccillus. A short rod shaped bacteria is called
coccobaccillus, a curved rod is called vibro, a curved rod long enough to form a spiral is called
spirillum, cocci that appears in pairs are called diplococci, long chain cocci is called
streptococci, cocci in clusters are staphylococci, cocci that divides forming cubical packet is
called sarcina. A long helical cell with a flexible cell wall and a unique mechanism of motility is
called Spirochete
3.4 FLAGELLA ARRANGEMENT IN BACTERIA CELL
From the figure above, there are five basic arrangement mode of flagella around a bacteria cell
and are labeled B - E. While some cells have no flagella others possesses one or more strands of
flagella. The figure above is explained below.
A- Atrichous: Not all bacteria are motile because they lack flagella and they are called
Atrichous Bacteria. Atrichous bacteria have no flagella. They move by means of gliding (e.g.,
Beggiatoa) or they don't move at all (e.g., cocci). Gliding bacteria will move only when they are
in contact with a solid plane. An example of gliding bacterium is Myxococcus. It decreases
surface tension at its posterior end by secreting a substance known as surfactant. The
difference in surface tension between the back and front of this bacterium causes it to glide.
B- Monotrichous or Polar Bacteria: Bacteria that possesses only one flagella at one end of
the cell (e.g., Vibro cholerae)
C- Lophotrichous Bacteria: They possesses tuft of flagella at one end of the cell (e.g.
Spirilla sp.)
D- Amphitrichous Bacteria: They posses single flagellum at both ends (e.g. Alcaligenes
faecalis)
D- Amphilophotrichous Bacteria: Bacteria cell that have tuft of flagella at both ends of
the cell.
E- Peritrichous: Bacteria with multiple flagella randomly surrounding the bacterial cell. All
the members of family Enterobacteriaceae, if motile, has peritrichous flagella. (e.g. Salmonella
typhi, Escherichia coli, Proteus sp.)
3.5 CULTURAL CHARACTERISTICS OF BACTERIA GROWTH ON SOLID AND

LIQUID MEDIA
Media Considerations
A culture medium must contain adequate nutrients to support bacterial growth. Minimally, this
would include organic compounds that can provide the building blocks necessary for cellular
reproduction. In many cases, predigested protein, such as hydrolyzed soy protein, serves this
purpose and will support the growth of many different bacteria.
Types of Media
● Complex/Undefined Media- This is a type of media that is not chemically defined,
formulations are generally referred to as complex media, to indicate that it is a mixture
with many components that are combined to support the growth of microorganism. It is
a media which has some complex ingredients, such as yeast extract or casein
hydrolysate, which consist of a mixture of many, many chemical species in unknown
proportions.
Undefined media are sometimes chosen based on price and sometimes by necessity –
some microorganisms have never been cultured on defined media
● Synthetic/ Defined Media- This is a medium in which all the chemicals used are known,
no yeast, animal, or plant tissue is present. A chemically defined medium is a growth
medium suitable for the culture of microbes or animal cells (including human) of which
all of the chemical components are known.
It represents the purest and most consistent cell culture environment. By definition
chemically defined media cannot contain either fetal bovine serum, bovine serum
albumin, or human serum albumin as these products are derived from bovine or human
sources and contain complex mixes of albumins and lipids.
● Selective Media- This is a type of media that contains additional substances such as an
antibiotic that may be selective for a particular type of bacteria by inhibiting most or all
other types and allowing growth of desirable bacteria, For example, if a microorganism
is resistant to a certain antibiotic, such as ampicillin or tetracycline, then that antibiotic
can be added to the medium in order to prevent other cells, which do not possess the
resistance, from growing. Example includes MacConkey agar, Thayer Martin's agar,
Lowenstein-Jensen media, Tellurite media (Tellurite inhibits the growth of most of the
throat organisms except diphtheria bacilli). Antibiotic may be added to a medium for
inhibition.
● Differential Media- This is a type of media that will have additional compounds that
permit an observer to distinguish among bacterial types based on differences in growth
patterns. This type of media uses the biochemical characteristics of a microorganism
growing in the presence of specific nutrients or indicators such as neutral red, phenol
red, eosin, or methylene blue added to the medium to visibly indicate the defining
characteristics of a microorganism. This type of media is used for the detection and
identification of microorganisms.
● Basal Media- This is a type of media that is used for growth of bacteria which do not
require further enrichment. Example includes Nutrient broth, nutrient agar, peptone
water. It can be used to culture Staphylococcus and Enterobacteriaceae.
● Enriched Media: This is a media type that is enriched usually by adding blood, serum or
egg, to support the growth of fastidious microoganism (microorganisms with complex or
particular nutrient requirement). Examples of enriched media includes blood agar and
Lowenstein-Jensen media. Streptococci grow in blood agar media.
● Indicator Media- A type of media with an indicator included in the medium. A particular
organism causes change in the indicator, e.g. blood, neutral red, tellurite. Example
includes Blood agar and MacConkey agar.
● Transport Media- These media are used when speciemen cannot be cultured soon after
collection. Example includes Cary-Blair medium, Amies medium, Stuart medium.
● Storage Media- Media used for storing the bacteria for a long period of time. Example
includes Egg saline medium, chalk cooked meat broth
Microbial Cultural Morphology

Microorganisms may show distinguishing gross morphologies when cultured on different
media. This macroscopic appearance of bacteria (characteristic growth patterns which can be
observed with the naked eye) is often used in their identification.
Selected strains of bacteria are inoculated on several types of media for the purpose of
observing and comparing their colonial growth characteristics.
By looking closely at the colonial growth on the surface of a solid medium, characteristics such
as surface texture, transparency, and the color or hue of the growth can be described. The
following three characteristics are readily apparent whether you’re looking at a single bacterial
colony or more dense growth, without the aid of any type of magnifying device.
Texture—Describes how the surface of the colony appears. Common terms used to describe
texture may include smooth, glistening, mucoid, slimy, dry, powdery, flaky etc.
Transparency—Colonies may be transparent (you can see through them), translucent (light
passes through them), or opaque (solid-appearing).
Color or Pigmentation—Many bacteria produces intracellular pigments which cause their

colonies to appear in a distinct color, such as yellow, pink, purple or red. Many bacteria do not
produce any pigment and appear white or gray.
Bacteriological Description of
Colonial Morphology
3.6 TEMPERATURE REQUIREMENTS OF BACTERIA
GRAPHICAL CHART OF TEMPERATURE REQUIREMENT OF

MICROORGANISMS (SOURCE: web2.mendelu.cz)
Bacteria can exist in a wide range of temperature from extreme cold to extreme hot. Each
prokaryote has a well defined upper and lower temperature limit within which it grows
optimally.
Optimum temperature is that temperature at which the microorganism readily multiplies most
rapidly.
Microorganisms found surviving under extreme conditions are called extremophiles, most of
these are members of domain Archaea. The temperature range of bacteria are as follows
Psychrophile:
● Cold loving bacteria with Optimum temperature of -5oC to 15oC.
● They are found in places that are permanently cold like Artic or Antarctic region.
● Example includes Psychrobacter arcticus, some Pseudomonas sp.
Psychrotroph:
● They are cold tolerant bacteria.
● Bacteria with Optimum temperature of 20oC to 30oC.
● They are important cause of food spoillage.
● Example includes Psychrobacter sp., Microbacterium sp.
Mesophile:
● Bacteria with Optimum temperature of 25oC to 45oC
● They are mostly disease causing bacteria that are adapted to grow within human body
temperature.
● Example includes Escherichia coli, Listeria monocytogenes, Staphylococcus aureus.
Thermophile:
● Bacteria with Optimum temperature of 45oC to 70oC
● They are found in hot springs and compost heaps.
● Example includes Thermus aquatics, Bacillus stearothermophilus.
Hyperthermophile:
● Bacteria with Optimum temperature of 70oC and above.
● These are usually members of Archaea.
● Example includes Pyrolobus fumarii, Pyrococcus furiosus.
3.7 OXYGEN REQUIREMENTS OF BACTERIA
1: Obligate aerobes need oxygen thus they gather at the top of the tube where the oxygen concentration is highest.
2: Obligate anaerobes are poisoned by oxygen, so they gather at the bottom of the tube where the oxygen concentration is
lowest.
3: Facultative anaerobes can grow with or without oxygen. They gather mostly at the top because aerobic respiration
generates more ATP than either fermentation or anaerobic respiration.
4: Microaerophiles need oxygen because they cannot ferment or respire anaerobically, but are poisoned by high
concentrations of oxygen. They gather in the upper part of the test tube but not the very top.
5: Aerotolerant organisms do not require oxygen and cannot utilise it even if present; they metabolise energy anaerobically.
Unlike obligate anaerobes, however, they are not poisoned by oxygen. They can be found evenly spread throughout the test
tube.
Like humans, some bacteria have absolute requirement for oxygen, others thrive in anaerobic
environment and many of these are killed if oxygen is present. Based on their oxygen
requirement, they can be separated into
Obligate Aerobe:
● They have absolute requirement for oxygen.
● They use oxygen to transform energy in aerobic respiration.
● Oxygen is used to metabolise substances, like sugars or fats, to obtain energy.
● With the exception of yeast, most fungi are obligate aerobes.
● Example includes Micrococcus sp., Mycobacterium tuberculosis, Nocardia asteroides.
Obligate Anaerobe:
● They cannot survive in oxygen presence infact are often killed in environment with trace
amount of oxygen.
● In other words they are poisoned by oxygen.
● They transform energy by fermentation or anaerobic respiration.
● Example includes Clostridium botulinum, Bacteroides fragilis, Clostridium perfringes.
Facultative Anaerobe:
● They grow better in oxygen presence but can also grow without oxygen. Facultative
means that the microorganism is flexible in it's oxygen requirement.
● They use aerobic respiration if oxygen is present and use fermentation in it's absence to
transform energy.
● Grow more rapidly when oxygen is present because aerobic respiration yield more ATP
than anaerobic.
● Cerrtain eukaryotes are also facultative anaerobes, including fungi (yeast) such as
Saccharomyces cerevisiae and many aquatic invertebrates such as Nereid (worm)
polychaetes.
● Example includes Escherichia colil, Staphylococcus aureus, Listeria monocytogenes.
Microaerophile:
● They need oxygen because they cannot ferment or respire anaerobically.
● However they require small amount of oxygen usually 2-10% for aerobic respiration.
● Many microaerophiles are also capnophiles, requiring an elevated concentration of
carbon dioxide (e.g. 10% CO2 in the case of Campylobacter species).
● Higher concentration of oxygen is poisonous to their growth and survival.
● Example includes Helicobacter pylori, Spirillum volutans.
Aerotolerant:
● They are indifferent to oxygen ie they can grow in it's presence or not although they do
not use it to transform energy.
● They are also called obligate fermenters.
● Example includes Streptococcus pyogene
3.8 TOXIC DERIVATIVE OF OXYGEN

Oxygen is not toxic itself but can be converted to a number of compounds that are highly toxic
(e.g. Superoxide O2 -) which is produced both as part of normal metabolism and as part of a
chemical reaction involving oxygen and light. Others such as hydrogen peroxide result from
metabolic process involving oxygen.
To survive in an environment containing toxic form of oxygen, cells must have enzymes that
convert this toxic compound to a non toxic form, For example, Superoxide dismutase which is
an enzyme degrades superoxide to produce hydrogen peroxide while catalase degrades
hydrogen peroxide to water (H2O) and oxygen (O2). Together, these two enzymes detoxifies the
toxic form of oxygen.
3.9 pH REQUIREMENT OF MICROORGANISMS

GRAPHICAL CHART OF pH REQUIREMENT OF MICROORGANISMS (SOURCE:
LUMENLEARNING.COM)
Every bacterial specie require certain range of pH values for survival. Most can live and
replicate within a range of pH 5(Acidic) to pH 8(Alkaline) and some can only survive at a neutral
pH of 7. Recall that acidity is a function of the concentration of hydrogen ions [H+] and is
measured as pH. Environments with pH values below 7.0 are considered acidic, whereas those
with pH values above 7.0 are considered basic or alkaline.
The optimum growth pH is the most favorable pH for the growth of an organism. The lowest pH
value that an organism can tolerate is called the minimum growth pH and the highest pH is the
maximum growth pH. These values can cover a wide range, which is important for the
preservation of food and to microorganisms’ survival in the stomach. For example, the
optimum growth pH of Salmonella spp. is 7.0–7.5, but the minimum growth pH is closer to 4.2.
Thus based on their pH requirement they can be separated into
Acidophile:
● An acidophile is an organism that can or must live in an acidic environment.
● An acidic environment is one that has a pH below 6.
● Acidophiles are able to live and thrive to a highly acidic environment, particularly at pH
2.0 or below.
● Natural niches where acidophiles can be found are volcanic areas, hydrothermal
sources, deep-sea vents, metal mining activities or in the stomachs of animals.
● Acidophiles are considered as an extremophile (organisms that grow and replicate
under extreme conditions of heat, pH or Carbon dioxide).
● Some acidophiles are adapted to an acidic environment because of a membrane system
that effectively pumps protons out of the intracellular space and consequently helps
keep the cytoplasm at or near a neutral pH. Other acidophiles with acidified cytoplasm
have proteins that can attain acid stability.
● Examples include Acetobacter aceti, i.e. a bacterium capable of producing acetic acid
through oxidizing ethanol, Acidobacterium capsulatum, Bryobacter aggregatus.
Neutrophile:
● A neutrophile refers to an organism that lives and thrives in an environment with a
neutral pH of about 6.5 to 7.5.
● Neutrophiles are adapted to live in an environment where the hydrogen ion
concentration is at equilibrium.
● They are sensitive to the concentration, and when the pH become too basic or acidic,
the cell's proteins can denature.
● Examples includes Escherichia coli, Staphylococci, Salmonella sp.
Alkaliphile
● A microorganism that grow best at pH between 8.0 and 10.5.
● Extreme alkaliphiles have adapted to their harsh environment through evolutionary
modification of lipid and protein structure and compensatory mechanisms to maintain
the proton motive force in an alkaline environment. For example, the alkaliphile Bacillus
firmus derives the energy for transport reactions and motility from a Na+ ion gradient
rather than a proton motive force.
● Many enzymes from alkaliphiles have a higher isoelectric point, due to an increase in
the number of basic amino acids, than homologous enzymes from neutrophiles.
● Example includes Bacillus firmus, Vibrio cholerae, the pathogenic agent of cholera,
grows best at slightly basic pH of 8.0; it can survive pH values of 11.0 but is inactivated
by the acid of the stomach.
Some neutrophile have adapted special mechanism that enables them grow in very low pH e.g.
Helicobacter pylori grows in the stomach where it causes ulcer. To maintain pH close to neutral,
Helicobacter pylori produces an enzyme urease which breaks down urea in the stomach to
produce carbondioxide (CO2) and ammonia (NH3). The ammonia neutralises the stomach acid
thus making it convenient for survival.
3.10 CHARACTERISTICS OF BACTERIA

● They are unicellular microscopic prokaryotes
● They have a size that ranges from 0.5 - 5 micrometer.
● They lack a well defined nucleus.
● They lack membrane bound organelles
● The cell wall is made up of a complex polymer called peptidoglycan.
● The peptidoglycan is responsible for the rigidity of the cell wall.
● Thick walled peptidoglycan bacteria are called Gram positive while thin walled
peptidoglycan bacteria are called Gram negative.
● They move using appendages extension from the cell called flagella.
● Some possesses capsule especially Gram negative bacteria while some don't.
● They reproduce asexually through binary fission.
● Enzymes for respiration and photosynthesis are located on the cytoplasmic membrane.
● Under unfavorable conditions they sure by becoming dormant, they form spores with a
thick protective coat around them which then split open in favorable condition.
● They undergo mutation occasionally (1000 times as much as eukaryotic gene) which
creates genetically different offspring.
● They are hybernated under low temperatures but killed in very high temperature.
NB: Archea have the same shape, size and appearance as a bacterial. Like bacteria they
multiply by binary fission and move by means of flagella, they have a rigid cell wall. Unlike
bacteria they do not posses peptidoglycan and their fatty acids are linked by ether bonds.
An interesting feauture of many members if their ability to grow in extreme environment in
which most organism cannot survive eg. They grow at extreme high temperature of 121 oC and
extreme low temperature in cold water of Antarctica, since are found at extreme salt
concentration of about ten times of that find in the sea.
3.11 DIFFERENCES BETWEEN ARCHAEA AND BACTERIA
ARCHEA BACTERIA
There's absence of peptidoglycan layer They posses peptidoglycan layer which

but presence of fatty acids in the cell wall. confers rigidity to the cell wall.
They are found in extreme environment. They are found in all environment, soil,
air, water, plant, animals etc
Their genes are more similar to Their genes are different from
eukaryotes. eukaryotes.
The membrane lipid are linked by ether The membrane lipid are linked by ester
bonds. bonds.
They posses several RNA polymerase. They posses one RNA polymerase.
Methionine serves as initiator tRNA Formylmethionine serves as initiator

tRNA.
Presence of protein similar to histone. They lack histone protein.
SIMILARITIES BETWEEN ARCHEA AND BACTERIA

● They both possesses same shape, size and appearance.
● They both multiply by binary fission.
● They both move by means of flagellla.
● They both have rigid cell wall.
● They both lack cellular organelles.
● They lack a well defined nucleus.
● They both have single circular chromosome.
● They both possesses nucleoid as they're genetic material.
● They both have 70s Ribosome.
● They both posses a cell membrane made up of phospholipid bilayer.
● They both possesses respiratory enzymes on the surface of the cell membrane.
● They can both exist as parasites of multicellular macroscopic entities such as plants and
animals.
● They both have initiator tRNA.
● They both possesses pilli why helps in adherence to a surface.
● They both posses RNA polymerase.
● They are both prokaryotes.
3.12 BACTERIAL DISEASES, CAUSATIVE AGENTS AND MODE OF

INFECTION
BACTERIAL DISEASES CAUSATIVE AGENTS MODE OF INFECTION
Tetanus Clostridium tetani ● Contamination of

wound by tetanus
spores.
● Cannot be
transmitted.
Botulism Clostridium botulinum ● Ingestion of

contaminated food
with bacterial spores.
● Inhalation of air
containing bacterial
spores (aerosol).
Anthrax Bacillus anthracis ● Direct contact with

infected person.
● Inhalation of aerosol.
Syphilis Treponema pallidum ● Sexual intercourse

with infected person
● Mother to foetus
though placenta.
● Direct contact of open
lesion with abraded
skin.
Thyphoid Salmonella typhi ● Ingestion of food,

water or milk
contaminated by fecal
matter (fecal - oral
route)
● Cannot be transmitted
Pneumonia Staphylococcus aureus ● Inhalation of aerosol

contaminated
surfaces (fomites).
Gastric ulcer Helicobacter pylori ● Fecal - oral route.
from person to
person.
Endocarditis Staphylococcus epidemis ● Direct contact with

infected person or
contaminated
surfaces.
● Vector route
● Inhalation of aerosol
Pharyngitis (sore throat) Streptococcus pyogene ● Direct contact with

infected person
Neonatal meningitis Escherichia coli ● Mother - foetus

through placenta.
Gastroenteritis Salmonella enterica ● Ingestion of

contaminated food
from person to
person.
Cholera Vibro cholerae ● Ingestion of

contaminated food,
water or milk.
from person to
person.
Tuberculosis Mycobacterium tuberculosis ● Direct contact with

droplet infection of
infected persons.
Gonorrhea Neiserria gonorrheae ● Sexual intercourse

Vaginitis Gardnerella vaginal ● Sexual intercourse
Leprosy Mycobacterium leprae ● Direct contact with

leprosy patients
● Mother to foetus
across placenta.
Yaws Treponema pertenue ● Vectors

infected person.
● Fomites.
Bacillary dysentery Shigella dysenteriae ● Fecal - oral route

from person to
person.
Food poisoning Staphylococcus aureus ● Ingestion of

contaminated food

Multiple Choice
1. A scientist observing a cell using a microscope noticed that the cell has a cell wall and
genetic material but lacks cellular organelles. What type of cell was observed (a)Plant
cell (b)Bacteria cell (c)Virus (d) Prokaryotic cell
2. The smallest known infectious agent in existence is (a)Bacteria (b)Prokaryotes
(c)Virus (d) Protozoa
3. A scientist observing a cell using a microscope noticed that the cell has a cell wall,
peptidoglycan layer and a genetic material but lacks cellular organelles. What type of
cell was observed (a)Plant cell (b)Bacteria cell (c)Virus (d) Prokaryotic cell
4. Microorganisms are ubiquitous meaning they can be found everywhere, but the ones
with the greatest distribution in nature is (a) Bacteria (b)Virus (c) Fungi
(d) Algae
5. Bacteria when first discovered by_____was called_________ (a) Louis Pasteur,
Animalcules (b) Antoni Van Leeuwenhoek, Animacules (c)Louis Pasteur,
Animayles (d) Antoni Van Leeuwenhoek Animayles
6. In an extreme environment of temperature above 130oC, microorganisms isolated are
likely to be (a)Bacteria (b)Viruses (c) Archea (d)Fungi
7. Food substance contaminated with Staphylococcus aureus when consumed is likely to
cause (a) Typhoid (b) Constipation (c) Food poisoning (d) Cholera
8. The following diseases are bacteria caused except (a) Measles (b) Leprosy
(c) Gonorrhoea (d) Tuberculosis
9. A complex molecule composed of alternating units of N-acetylmuramic acid (NAM) and
N-acetylglucosamine (NAG) which confers rigidity to bacteria cell wall is (a) Chitin
(b) Cellulose (c) Muranose (d) Peptidoglycan
10. A bacteria cell with zero motility functions and ability is best considered (a) Atrichous
(b) Peritrichous (c) Amphitrichous (d) None of the above
11. During a field work on a pH environment of 8.0 - 10.5, some microorganism were
observed to flourish well at such pH, such organisms are best suspected to be (a)
Acidophile (b) Alkaliphile (c) Neutrophile (d) Mesophile
12. A patient was diagnosed with cholera after several laboratory tests. What is the likely
causal organism of this disease is (a) Vibro cholera (b) Salmonella cholera
(c) Salmonella cholerae (d) Vibro cholerae
13. A type of media that is not chemically defined but contains mixture of compounds in
unknown proportion is called (a) Complex media (b) Basal Media (c)
Differentiatial Media (d) Synthetic Media
14. A scientist isolated and incubated a strain of bacteria, upon observation using a
microscope, he observed that they were rod shaped bacteria and are more likely
(a) Coccobaccillus (b) Coccus (c) Baccillus (d) Spirochete.
15. A small circular extra-chromosomal DNA structure capable of autonomous replication is
(a) Plasmid (b) Plastid (c) Plasmolema (d)Pili
16. A bacteria cell with a single flagella for motility functions is best considered (a) Atrichous
(b) Peritrichous (c) Amphitrichous (d) Monotrichous
17. A nutrient media contains a Colony of microorganisms. This colony was observed dead
after exposure to air. The colony was made up of_____microorganisms (a) Anaerobic
(b)Aerobic (c)Facultative anaerobic (d) Micro aerophiles
microscope, he observed that they were short rod shaped bacteria and are more likely
(a) Coccobaccillus (b) Coccus (c) Baccillus (d) Spirochete.
19. All of the following are similarities between Archae and Bacteria except (a) Both
possesses 70s ribosome (b) Both possesses peptidoglycan layer (c) Both possesses
nucleoid as the genetic material (d) Both multiply through binary fission.
20. All of the following are differences between Archae and Bacteria except (i) Both
possesses 70s ribosome (ii) Both possesses peptidoglycan layer (iii) Both possesses
nucleoid as the genetic material (iv ) Both multiply through binary fission.
(a) I, ii, iv (b)I, ii, iii, iv (c)I, iii, iv (d) iii, iv
21. The amino acid which serves as initiator tRNA in Archae is (a)
Formylmethionine (b) Tryptophan (c) Aspartic acid (d) methionine
22. A type of media that contains additional substances such as an antibiotic that inhibits
most or all other types of bacteria and allows growth of desirable bacteria is known as
(a) Differential media (b)Enriched media (c) Selective media (d)Complex media
microscope, he observed that they were curved rod shaped bacteria and are more likely
(a) Coccobaccilus (b) Coccus (c) Baccillus (d) Vibro
24. The amino acid which serves as initiator tRNA in bacteria is (a)
Formylmethionine (b) Tryptophan (c) Aspartic acid (d) methionine
25. During a field work on a pH environment of 2.0 and below , some microorganism were
observed to flourish well at such pH, such organisms are best suspected to be (a)
Acidophile (b) Alkaliphile (c) Neutrophile (d) Mesophile
26. They grow better in oxygen presence but can also grow without oxygen. This description
best fits (a) Anaerobes (b) Facultative Anaerobes (c)Aerobes (d) None of the
above.
27. Bacteria with Optimum temperature of 25oC to 45oC and are implicated as disease
causing bacteria are
(a) Thermophile (b)Psychrophile (c) Psychrotroph (d) Mesophile
28. Bacteria with Optimum temperature of 20oC to 30oC and are mostly important causes of
food spoillage are
(a) Thermophile (b)Psychrophile (c) Psychrotroph (d) Mesophile
29. A microorganism is said to be fastidious if (a) It grows tightly
fastened to the media (b) It has a fast exponential growth rate (c)it has a complex or
particular nutrient requirement(d) It can be easily and cheaply culture in the laboratory
with all conditions being suitable.
30. A bacteria cell with a single flagella each at both ends used for motility functions is best
considered (a) Atrichous (b) Peritrichous (c) Amphitrichous (d)
Monotrichous
31. The causal organism of Anthrax disease in livestock is (a)Salmonella enteric (b)Bacillus
anthracis (c)Salmonella anthracis (d) Vibro cholera
32. Identity the correct statement (a) To kill a bacteria cell, subject it to temperature
below 0oC (b) To kill a bacteria cell, subject it to an environment with no
oxygen availability (c)To kill a bacteria cell, subject it to an
environment with high oxygen availability (d) To kill a
bacteria cell, subject it to extremely high temperature
33. A bacteria with multiple flagella randomly surrounding the bacteria cell used for motility
functions is best considered (a) Atrichous (b) Peritrichous
(c) Amphitrichous (d) Monotrichous
34. A media type that is enriched usually by adding blood, serum or egg, to support the
growth of fastidious microoganism is (a) Differential media (b)Enriched media
(c) Selective media (d) Complex media
35. The bacteria cell structure that provides bouyancy to cell is (a)Plasmid
(b)Pilli (c) Ribosomes (d) Gas vesciles
36. Cocci bacteria that divides forming cubical packet is called (a) sarcina (b) Spirochete
(c) Spirillum (d)None of the above
37. David sustained a deep foot injury after colliding with a sharp stone while playing
soccer. The wound got contaminated producing a foul smell. What possible condition is
this (a)Filiriasis (b) Tetanus (c) Hepatitis (d) Yaws
38. With reference to the question above select the correct option (a) The
disease above is highly contagious (b) The disease above is mildly contagious (c)
The disease above is slowly contagious (d) The disease above is not contagious
39.
The correct name of the flagella arrangement above is. (a)peritrichous & lophotrichous.
(b)amphitrichous & atrichous. (c) lophotrichous & amphilophotrichous.
(d)Polar & amphilophotrichous.
40. Which of the following bacteria diseases are sexually transmitted (i) Gonorrhoea
(ii) AIDS (iii) Syphilis (iv) Vaginitis
(a)i, ii, iii (b) i, ii, iv (c) i, iii, iv (d) i ii, iii, iv
41. Which of the following bacteria diseases are transmitted by the consumption of
contaminated food (i) Gastroenteritis (ii) Cholera (iii) Polio (iv) Food
poisoning

Theory
1. With the aid of a diagram, explain the function of the structures of a bacteria cell.
2. Accurately define the following terms (i)Spirochete (ii) Bactericidal (iii)Fecal - oral
route (iv)Sterilization (v) Fastidious bacteria (vi)Basal Media (vii) Facultative
anaerobes (viii) Amphilophotrichous (ix)Synthetic Media (x) Coccobacillus
3. Explain with relevant examples where necessary, the temperature requirement of

bacteria.
4. Enumerate 5 Differences and similarities each between Archae and Bacteria
5. Explain with relevant examples where necessary, the flagella arrangement of bacteria.
6. The diagram below depicts the growth pattern of bacteria in the presence of oxygen.
Using the diagram above, explain oxygen requirement of bacteria citing relevant
example in each case.
7. Appropriately fill the table below.
Tetanus ●
Clostridium botulinum ●
Treponema pallidum ●
Thyphoid ●
Helicobacter pylori ● Fecal - oral route.

from person to
person.
Pharyngitis (sore throat) ●
Vibro cholerae ●
Tuberculosis ●
Gonorrhoea ●
Mycobacterium leprae ●
8. Enumerate 10 differences and 5 similarities between bacteria and virus
9. Differentiate between the following pair (i) Synthetic

& Non-synthetic media (ii) Selective & Enriched media (iii) Basal & Storage
media (iv) Neutrophile & Alkaliphile (v) Nucleus and Nucleoid
10. Explain how Helicobacter pylori an infectious agent tends to survive in the acidic
environment of the human stomach. (b) State 3 preventive
measures to be taken against this infectious agent
11. Explain the phrase "Cultural Morphology of Microorganisms" (b) Describe the
following terms as it applies to the cultural Morphology of microorganisms
(i) Texture (ii) Transparency (iii)
Pigmentation
12. All bacteria are disease causing entities TRUE or FALSE (b) Justify
your answer
13. State 5 Economic importance of bacteria

1. D
2. C
3. B
4. A
5. B
6. C
7. C
8. A
9. D
10. A
11. B
12. D
13. A
14. C
15. A
16. D
17. A
18. A
19. B
20. C
21. D
22. C
23. D
24. A
25. A
26. B
27. D
28. C
29. C
30. C
31. B
32. D
33. B
34. B
35. D
36. A
37. B
38. D
39. C
40. B
THEORY
1. Make reference to Chapter3.2
2. Spirochete is a long helical cell with a flexible cell wall and a unique mechanism of
motility.
Bactericidal is a substance of chemical complexity that kills bacteria out-rightly.
Fecal - Oral route is a term which explains the mode of transmission of causal organism
of infectious disease, when particles from fecal matter of infected person ends up in the
mouth of a healthy individual.
Sterilization is a process that kills out-rightly all forms of microbial life and biological
agents on the surface of an inanimate object.
Fastidious bacteria are bacteria that require special or complex growth requirement.
Basal media is a type of media that is used for growth of bacteria which do not require
further enrichment.
Facultative Anaerobes are microorganisms that grow better in the presence of oxygen
using aerobic respiration but in it's absence also grow using fermentation to transform
energy.
Amphilophotrichous is a type of flagella arrangement in bacteria characterised with tuft

of flagella at both ends of the cell.
Synthetic Media is a media in which all the chemicals used are known, no yeast, animal,
or plant tissue is present.
Coccobacillus is a short rod shaped bacteria.

8.
BACTERIA VIRUSES
They are prokaryotes They are particles
They have a nucleoid which contains the They have a genome which is the genetic
genetic material material
They have a rigid cell wall which confers Some have envelope which confers
protection. protection.
Their cell wall is made up of Their envelope is made up of portions of

peptidoglycan layers. their host cell membrane.
Following the cell wall is a semi- Following the envelope is the capsid.
permeable membrane.
They are made up of 70s ribosome They lack ribosomes.
They are unicellular parasites They are acellular parasites
They are majorly heterotrophs with few They are neither heterotrophs nor
autotrophs. autotrophs because they are regarded as
nonliving things.
They have a diameter of 1 - 10 They have a diameter of 20 - 300

micrometer nanometers
They can be cultured in the lab using They can only be cultured using cell
simple nutrient media. culture or embryonated egg.
Similarities
● They are both microscopic entities that cannot be seen with the naked eye.
● They are parasitic to plants and animals.
● They can both attach or adhere to a surface using specialized structures.
● They both possesses a genetic material useful in replication.
● They can both be cultured invitro in the laboratory
● They can both be killed under extremely high temperatures
● They can both be used in bioterrorism.
10. Helicobacter pylori grows in the stomach where it causes ulcer. To maintain pH close to
neutral, Helicobacter pylori produces an enzyme urease which breaks down urea in the
stomach to produce carbondioxide (CO2) and ammonia (NH3). The ammonia neutralises the
stomach acid thus making it convenient for survival in the acidic environment.
(b) Fecal matter should be properly and aseptically disposed and eliminated.
Avoid the consumption of under-cooked and unwashed food or fruit.
Effective personal hygiene should be practiced and maintained always.
12. TRUE. All bacteria can cause disease if present in a susceptible or immuno compromised
host and/or present in organs or tissues that doesn't serve as their normal habitat. E.g
Staphylococcus aureus the normal microflora of the skin may not cause diseases there but if
found inside the body of an immuno compromised host can cause a wide range of diseases and
havoc.
13. Bacteria are useful agents in bioremediation e.g. Pseudomonas putida
They are important causal organism of many human and animal diseases such as Tuberculosis
(Mycobacterium tuberculosis) and Anthrax (Bacillus anthracis) respectively.
Due to their rapid reproduction rate, they are used in genetic engineering for the production of
medically important product such as insulin, interferon, enzymes, hormones etc.
They facilitate the growth of agricultural crops using their ability to fix atmospheric nitrogen in
the soil, thereby making nutrients available for plant use.
They can be modified to produce vaccines which confers active immunity against diseases. E.g
leprosy vaccine
They can also be used to genetically modify plants so they become resistant to insect attack and
viral diseases.
They also facilitate food spoillage. E.g Micrococcus sp
They facilitate milk fermentation useful in yoghurt production and as probiotics. E.g Lactic acid
bacteria.
CHAPTER 4
4.1 FUNGI
Fungi are members of the group of eukaryotic organisms which includes microorganisms such
as yeasts and molds as well as the more familiar mushrooms. These organisms are classified as
kingdom Mycota, which is separate from the other eukaryotic life kingdoms of plants and
animals.
The fungi comprise a diverse group of organisms that are heterotrophic and typically saprozoic.
In addition to the well-known macroscopic fungi (such as mushrooms and molds), many
unicellular yeasts and spores of macroscopic fungi are microscopic. For this reason, fungi are
included within the field of microbiology.
Fungi are important to humans in a variety of ways. Both microscopic and macroscopic fungi
have medical relevance, with some pathogenic species that can cause mycoses (illnesses caused
by fungi). Some pathogenic fungi are opportunistic, meaning that they mainly cause infections
when the host’s immune defenses are compromised and do not normally cause illness in
healthy individuals. Fungi are important in other ways. They act as decomposers in the
environment, and they are critical for the production of certain foods such as cheeses. Fungi are
also major sources of antibiotics, such as penicillin from the fungus Penicillium.
STRUCTURE OF A FUNGAL CELL (biologydiscussion.com)
A characteristic that places fungi in a different kingdom from plants, animals, bacteria, and
some protists is the presence of chitin in their cell walls. They are responsible for
decomposition and nutrient cycling through the environment.
They are found in a range of habitat from terrestrial to aquatic in dark and moist places, on
substratum and on dead organic matter.
Fungi, once considered plant-like organisms, are more closely related to animals than plants.
Fungi are not capable of photosynthesis: they are heterotrophic because they use complex
organic compounds as sources of energy and carbon. Some fungal organisms multiply only
asexually, whereas others undergo both asexual reproduction and sexual reproduction with
alternation of generations. Most fungi produce a large number of spores, which are haploid
cells that can undergo mitosis to form multicellular, haploid individuals. Like bacteria, fungi play
an essential role in ecosystems because they are decomposers and participate in the cycling of
nutrients by breaking down organic and inorganic materials to simple molecules.
The somatic body of fungi is unicellular or multicellular, when multicellular, it's composed of
profusely branched interwoven thread like structure called Hyphae, the mass of collection of
Hyphae is called Mycelium.
Hyphae may be continuous or interrupted by crosswalls called septa which divides the Hyphae
into compartments thus Hyphae may be septate or aseptate.
Aseptate Hyphae - Hyphae with no crosswalls and thus they are ceonocytic i.e. containing
many nuclei.
Septate Hyphae- Hyphae with crosswalls forming compartments thus they are aceonocytic
i.e. one nuclei per compartment.
The study of fungi is called Mycology.
4.2 Key Terms in Mycology.

Ascomycota: a taxonomic division within the kingdom Fungi; those fungi that produce spores
in a microscopic sporangium called an ascus
Heterotrophic: organisms that use complex organic compounds as sources of energy and
carbon.
Glucan: any polysaccharide that is a polymer of glucose
Ergosterol: the functional equivalent of cholesterol found in cell membranes of fungi and some
protists, as well as, the steroid precursor of vitamin D2
Mycelium: the vegetative part of any fungus, consisting of a mass of branching, threadlike
hyphae, often underground
Hypha: a long, branching, filamentous structure of a fungus that is the main mode of
vegetative growth
Septum: cell wall division between hyphae of a fungus
Thallus: vegetative body of a fungus that cannot be differentiated into root, stem and leaves.
Saprophyte: any organism that lives on dead organic matter, as certain fungi and bacteria
Decomposer: any organism that derives energy by feeding on dead and decaying organic
matter, as certain fungi.
Chitin: a complex polysaccharide, a polymer of N-acetylglucosamine, found in the exoskeletons
of arthropods and in the cell walls of fungi; thought to be responsible for some forms of asthma
in humans
Homothallic: male and female reproductive structures are present in the same plant or fungal
mycelium
Gametangium: an organ or cell in which gametes are produced that is found in many
multicellular protists, algae, fungi, and the gametophytes of plants
Spore: a reproductive particle, usually a single cell, released by a fungus, alga, or plant that
may germinate into another
Sporangium: a case, capsule, or container in which spores are produced by an organism
Sporangiophore: a long firm filamentous structure that holds the sporangium at it's peak.
Karyogamy: the fusion of two nuclei within a cell
plasmogamy: stage of sexual reproduction joining the cytoplasm of two parent mycelia
without the fusion of nuclei
4.3 Fungal Association

Mycorrhiza: F ungi often interact with other organisms, forming beneficial or mutualistic
associations. For example most terrestrial plants form symbiotic relationships with fungi. The
roots of the plant connect with the underground parts of the fungus forming mycorrhizae.
Through mycorrhizae, the fungus supplies the past root with water and the plant supplies the
fungus with nutrients thus nutrient and water is being exchanged which greatly aids the survival
of both specie.
Lichen: Alternatively, lichens are an association between a fungus and its photosynthetic
partner (usually an alga). The fungus traps, absorbs and supplies the algae partner with water
while the algae being photosynthetic supplies the fungus with it's self synthesised nutrients.
They are often found as white or yellow patches on old walls, bricks, rocks etc.
4.4 Fungal Classification
Kingdom Fungi (Mycota) is divided into two namely:
● Myxomycota (plasmodium or pseudo fungi)
● Eumycota: subdivided into Mastigomycotina, Zygomycotina, Ascomycotina,
Basidiomycotina, Deuteromycotina.
Myxomycotina
● They behave like animals at vegetative stage (at birth) because they lack cell walls, and
behave like fungi at reproductive stage.
● They are found in damp and shady places.
● The body of some members is known as plasmodium (a naked slimy protoplasmic mass
with many nuclei)
● Plasmodium moves with pseudopodia in an amoeboid fashion.
● It feeds by engulfing food materials (phagocytosis)
● Reproduce via fragmentation, zoospores and other asexual spores.
● Example includes Mycetozoa, Myxomycota etc
Eumycota: Further subdivided into five classes

Mastigomycotina:
● They are zoosporic fungi ie they produce zoospores as a means of asexual reproduction.
● Mainly found in aquatic or damp environment.
● They are ceonocytic myceliumi.e presence of many scattered nuclei.
● They have aseptate hyphae.
● Reproduce sexually through conjugation.
● Reproduce asexually through zoospores.
● They produce flagellated cells during their lifetime.
● May bear rhizoids.
● Mostly, filamentous and having coenocytic mycelium.
● Show centric nuclear division.
● Perfect state of spores is typically oospores.
● They are further subdivided into Chytridiomycetes, Hyphochytridiomycetes and
Oomycetes.
● Example includes Physoderma maydis, Olpidium brassicae etc.
Zygomycotina:
● There are approximately 1060 known species.
● They are mostly terrestrial in habitat, living in soil or on decaying plant or animal
material.
● Some are parasites of plants, insects, and small animals, while others form symbiotic
relationships with plants.
● Zygomycete hyphae may be coenocytic, forming septa only where gametes are formed
or to wall off dead hyphae.
● They have aseptate hyphae.
● Asexually, they produce through non motile spores called sporangiospore which are
usually blown by air.
● Sexually, they reproduce by conjugation through zygospore.
● They are further subdivided into Mucoromycotina, Kickxellomycotina,
Entomophthoromycotina, and Zoopagomycotina.
● Example includes Mucor, Rhizopus, Pirella, Gamsiella, Linderina, Endogone etc.
ZYGOMYCETES LIFE CYCLE
Ascomycotina:
● Commonly known as the sac fungi or ascomycetes.
● It is the largest phylum of Fungi, with over 64,000 species.
● The defining feature of this fungal group is the "ascus" (meaning "sac" or "wineskin")
which is a microscopic sexual structure in which nonmotile spores, called ascospores,
are formed.
● Sexual reproduction takes place through a process called gametangial corpulation.
● Some species of the Ascomycota are asexual, meaning that they do not have a sexual
cycle and thus do not form asci or ascospores.
● Yeast which is a common example reproduce asexually by budding while others
reproduce through non motile spores such as conidia.
● Many ascomycetes are pathogens, both of animals, including humans, and of plants.e.g
rice blast, apple scab, Candida albicans etc.
● They have septate hyphae and thus are aceonocytic.
● An important respiratory pathogen is the fungus Histoplasma capsulatum which is
associated with birds and bats in the Ohio and Mississippi river valleys.
● Candida albicans, the most common cause of vaginal and other yeast infections, is also
an ascomycete fungus; it is a part of the normal microbiota of the skin, intestine, genital
tract, and ear.
● Example includes Saccharomyces cerevisea (yeast), Penicillium notatum, Aspergillus
niger, Candida albicans etc.
ASCOMYCETES LIFE CYCLE
Basidiomycotina:
● They are regarded as club fungi.
● Commonly referred to as higher fungi.
● These are filamentous fungi composed of hyphae (except for basidiomycota-yeast).
● They reproduce sexually via the formation of specialized club-shaped end cells called
basidia that normally bear external meiospores (usually four).
● These specialized spores are called basidiospores.
● Basidiospores are produced in basidium.
● Asexually they reproduce through non motile spores e.g telospores.
● Some Basidiomycota are obligate asexual reproducers. Basidiomycota that reproduce
asexually can typically be recognized as members of this division by gross similarity to
others.
● Example include Mushroom, Toadstool, Puffballs, Stinkhorns, Bracket fungi, Smuts,
Bunts, Rusts, Mirror yeasts etc.
BASIDIOMYCETES LIFE CYCLE
Deuteromycotina
● There are about 25,000 species that have been classified in the deuteromycotaand
many are basidiomycota or ascomycota anamorphs.
● Other, more informal names besides Deuteromycota ("Deuteromycetes") and fungi
imperfecti are anamorphic fungi, or mitosporic fungi, but these are terms without
taxonomic rank.
● They are regarded as fungi imperfecti because they lack sexual stage.
● Known for their asexual stage only called anamorphic.
● They produce their spores asexually, in the process called sporogenesis.
● Sexual reproduction is not known.
● Example includes Alternaria, Colletotrichum, Trichoderma etc.
4.5 FUNGAL NUTRITION

Like animals, fungi are heterotrophs: they use complex organic compounds as a source of
carbon, rather than fix carbon dioxide from the atmosphere as do some bacteria and most
plants.
Fungi are considered as the richest set of microbes in terms of enzyme production, as they
produces a wide range of enzyme used to carry out extra cellular digestion.
In addition, fungi do not fix nitrogen from the atmosphere. Like animals, they must obtain it
from their diet. However, unlike most animals, which ingest food and then digest it internally in
specialized organs, fungi perform these steps in the reverse order: digestion precedes ingestion.
First, exoenzymes are transported out of the hyphae, where they process nutrients in the
environment. Then, the smaller molecules produced by this external digestion are absorbed
through the large surface area of the mycelium. As with animal cells, the polysaccharide of
storage is glycogen rather than the starch found in plants.
Fungi are mostly saprobes (saprophyte is an equivalent term): They obtain their nutrients from
dead or decomposing organic matter, mainly plant material. Fungal exoenzymes are able to
break down insoluble polysaccharides, such as the cellulose and lignin of dead wood, into
readily-absorbable glucose molecules. The carbon, nitrogen, and other elements are thus
released into the environment. Because of their varied metabolic pathways, fungi fulfill an
important ecological role and are being investigated as potential tools in bioremediation.
4.6 Fungal Reproduction

Fungi reproduce sexually and/or asexually. Perfect fungi reproduce both sexually and asexually,
while imperfect fungi reproduce only asexually (by mitosis).
In both sexual and asexual reproduction, fungi produce spores that disperse from the parent
organism by either floating on the wind or hitching a ride on an animal. Fungal spores are
smaller and lighter than plant seeds. The giant puffball mushroom bursts open and releases
trillions of spores. The huge number of spores released increases the likelihood of landing in an
environment that will support growth.
Asexual Reproduction
Fungi reproduce asexually by fragmentation, budding, or producing spores. Fragments of
hyphae can grow new colonies. Mycelial fragmentation occurs when a fungal mycelium
separates into pieces with each component growing into a separate mycelium. Somatic cells in
yeast form buds. During budding (a type of cytokinesis), a bulge forms on the side of the cell,
the nucleus divides mitotically, and the bud ultimately detaches itself from the mother cell.
The most common mode of asexual reproduction is through the formation of asexual spores,
which are produced by one parent only (through mitosis) and are genetically identical to that
parent. Spores allow fungi to expand their distribution and colonize new environments. They
may be released from the parent thallus, either outside or within a special reproductive sac
called a sporangium.
Sexual Reproduction
Sexual reproduction introduces genetic variation into a population of fungi. In fungi, sexual
reproduction often occurs in response to adverse environmental conditions. Two mating types
are produced. When both mating types are present in the same mycelium, it is called
homothallic, or self-fertile. Heterothallic mycelia require two different, but compatible, mycelia
to reproduce sexually.
Fungi reproduce sexually either through cross- or self-fertilization. Haploid fungi form hyphae
that have gametes at the tips. Two different mating types (represented as “+ type” and “–
type”) are involved. The cytoplasms of the + and – type gametes fuse (in an event called
plasmogamy), producing a cell with two distinct nuclei (a dikaryotic cell). Later, the nuclei fuse
(in an event called karyogamy) to create a diploid zygote. The zygote undergoes meiosis to form
spores that germinate to start the haploid stage, which eventually creates more haploid
mycelia. Depending on the taxonomic group, these sexually produced spores are known as
zygospores (in Zygomycota), ascospores (in Ascomycota), or basidiospores (in Basidiomycota).
At this stage, spores are disseminated into the environment.
4.7 Characteristics of Fungi

Fungi have well-defined characteristics that set them apart from other organisms. Most
multicellular fungal bodies, commonly called molds, are made up of filaments called hyphae.
Hyphae can form a tangled network called a mycelium and form the thallus (body) of fleshy
fungi.
In contrast to molds, yeasts are unicellular fungi.
● They are eukaryotic achlorophyllus organisms
● Most are multicellular while few are unicellular.
● Fungi are heterotrophic: they use complex organic compounds as sources of energy and
carbon, not photosynthesis.
● Fungi multiply either asexually, sexually, or both.
● The majority of fungi produce spores, which are defined as haploid cells that can
undergo mitosis to form multicellular, haploid individuals.
● Fungi interact with other organisms by either forming beneficial or mutualistic
associations (mycorrhizae and lichens ) or by causing serious infections.
● They are majorly decomposers – the best recyclers around, and exist as saprophytes.
● Most multicellular (hyphae) – some unicellular (yeast)
● They are non-motile
● They have complex cellular organisation.
● Cell walls made of chitin (kite-in) instead of cellulose like that of a plant
● Are more related to animals than plant kingdom
● They are made up of a thallus body.
● They Lack true roots, leaves and stems
● Absorptive heterotrophs -Digest food externally and then absorb it
● They do not depend on light.
● Can occupy dark habitats
● Can grow in any direction
● Possesses strong absorptive filaments
● They can penetrate the interior of a substrate with absorptive filaments.
● Store food as glycogen just like animals.
● Whereas animals have cholesterol in their cell membranes, fungal cell membranes have
different sterols called ergosterols. Ergosterols are often exploited as targets for
antifungal drugs.
● Body of fungus made of tiny filaments or tubes called hyphae .
● Contain cytoplasm and nuclei (more than 1)
● Each hyphae is one continuous cell
● Cell wall made of chitin
● A tangled mess of hyphae is called mycelium
● Rhizoids are root-like parts of fungi that anchor them to the substrate (whatever they
are bonding to)
● Mycelium increase the surface area of the fungi to absorb more nutrients.
4.8 FUNGAL DISEASES, CAUSATIVE AGENTS AND MODE OF INFECTION

FUNGAL DISEASES CAUSAL AGENTS
Athletic foot Trichophyton

rubrum
Vaginal Candidiasis Candida albicans
Ringworm Epidermophyton sp
Aspergillosis Aspergillus niger
Pneumocystis pneumomia Pneumocystis jirovecii.
Mucormycosis Rhizopus oryazae
Talaromycosis Talaromyces marneffei
Jock itch Microsporum sp
Blastomycosis Blastomyces dermatitidis
Valley Fever Coccidioides immitis

Multiple Choice
1. Kingdom Mycota is exclusively reserved for a particular type of microorganisms and they
are (a) Bacteria (b) Archae (c) Fungi (d) Protozoa
2. Fungi are best described as (a) Unicellular eukaryotic chlorophyllous

heterotrophs. (b) Multicellular eukaryotic achlorophyllus
heterotrophs. (c) Unicellular prokaryotic chlorophyllus autotrophs
(d) Multicellular prokaryotic chlorophyllous autotrophs.
3. The somatic body of fungi when multicellular is composed of profusely branched

interwoven thread like structure called (a)Mycelium (b) Mycelia (c) filaments
(d) Hyphae
4. Root-like parts of fungi that anchors them to substrates can be suitably referred to as
(a)Rhizoid (b) Stipe (c) Cone (d) Root
5. Similarities between fungi and animals includes which of the following (i) Store
food as glycogen (ii) Presence of chitin in their cell wall (iii)Eukaryotic cell
structure (iv) Heterotrophic mode of nutrition
6. A complex polysaccharide, a polymer of N-acetylglucosamine, found in the exoskeletons

of arthropods and in the cell walls of fungi; thought to be responsible for some forms of
asthma in humans is (a) Hydrostatic fluid (b) Skeletal protein (c) Cellulose
(d)Chitin
7. A case, capsule, or container in which spores are produced by an organism (a)
Sporangium (b) Sporangiophore (c) Sori (d) None of the above
8. The vegetative part of any fungus, consisting of a mass of branching, threadlike hyphae
is called (a)Ascus (b) Stipe (c) Rhizoid (d) Mycelium
9. Aseptate Hyphae is best described as (a) Hyphae with no crosswalls and thus they are
ceonocytic (b) Hyphae with crosswalls and thus they are ceonocytic
(c) Hyphae with crosswalls and thus they are not ceonocytic (d) Hyphae
with no crosswalls and thus they are not ceonocytic
10. Similarities between fungi and plants includes which of the following (i) Presence
of cellulose on cell wall (ii) Presence of chitin in their cell wall (iii)Eukaryotic cell
structure (iv) Autotrophic mode of nutrition
(a)i, ii only (b) i, ii, iii (c) iii only (d) i ii, iii, iv
11. The implication of chlorophyllusity of an organism is that (a) Such

organism becomes self dependent (b) Such organism adapts
to heterotrophism (c) Such organism adapts to parasitism (d)
Such organism adapts to autotrophism
12. The cell membrane of fungi contains (a) Chitin (b) Steroids (c) Ergosterol (d)
Cholesterol
13. A reproductive particle, usually a single cell, released by a fungus, alga, or plant that
may germinate into another (a) Sporangium (b) Sporangiophore (c)
Spores (d) Sori
14. Diseases caused by fungi are called (a) Fungal diseases (b) Mycology (c)
Mycoses (d) Fungal infections
15. During karyogamy, which of the following statement is true (a)the haploid
nuclei fuse to form a diploid zygote nucleus. (b)the diploid nuclei fuse to
form a diploid zygote nucleus. (c)the haploid nuclei fuse to form a
haploid zygote nucleus. (d) the diploid nuclei fuse to form a haploid zygote
nucleus.
16. The most common mode of asexual reproduction in fungi is through the formation of
(a) Asexual filaments (b) Asexual spores (c) Asexual buds (d)Asexual gametes
17. Which of the following statement is correct (a) Fungi
imperfecti reproduces only asexually (b) Fungi imperfecti reproduces only sexually
(c) Fungi imperfecti reproduces through both sexually and asexually (d) Fungi
imperfecti reproduces via dissemination and dissertation of spores.
18. A long firm filamentous structure that holds the sporangium at it's peak is (a) Sori
(b) Sporangiophore (c) Spores (d) None of the above
19. The similarity between fungi and bacteria is (a) They are both exhibit sexual and
asexual reproduction (b) They are both eukaryotic (c) They both prokaryotic (d)
They are both heterotrophic
20. Septate Hyphae is best described as (a) Hyphae with no crosswalls and thus they are
ceonocytic (b) Hyphae with crosswalls and thus they are ceonocytic
(c) Hyphae with crosswalls and thus they are not ceonocytic (d) Hyphae
with no crosswalls and thus they are not ceonocytic
21. The implication of achlorophyllusity of an organism is that (a) Such

organism becomes self dependent (b) Such organism adapts
to heterotrophism (c) Such organism adapts to parasitism (d)
Such organism adapts to autotrophism
22. The similarity between fungi and virus is (a) They are both eukaryotic (b)
They are both heterotrophic (c) They both prokaryotic (d) They both possesses a
genetic material
23. The class of fungi regarded as club fungi is (a) Ascomycota (b)
Basidiomycotina (c) Zygomycotina (d)
Deuteromycotina
24. Which group of fungi appears to be associated with the greatest number of human
diseases? (a) Ascomycota (b)
Deuteromycotina
25. The class of fungi regarded as sac fungi is (a) Ascomycota (b)
Deuteromycotina
26. The joining of cytoplasm of two parent mycelia without the fusion of nuclei is called
(a) Karyogamy (b) Isogamy (c) Plasmogamy (d) Heterogamy
27. The most diversified microbes in nature are (a) Fungi (b) Bacteria (c) Algae (d)
Viruses
28. Fungi are considered the richest microbial life because fungi (a) Possesses
a rapid reproduction rate (b) Are able to produce a wide range of enzymes
(c) Are the most diversified microbe in nature (d) Are found everywhere.
29. The product of the fusion of two nuclei in a fungal cell is (a) Embryo (b) Two daughter
nuclei in one cell (c) Zygote (d) None of the above
30. The causal organism of Ring worm is (a) Dermatophytes (b) Candida albicans (c)
Aspergillus niger (d) Epiphytes
31. Symbiotic relationship between root of terrestrial plants with fungi is referred to as
(a) Lichen (b) Mycorhhiza (c) Mycorrhiza (d) Rhizopus
32. Asexual reproduction of fungi includes all the following except (a) Budding (b) Fusion
of gamete (c) Fragmentation (d) Production of spores
33. Unicellular fungi are called (a) Molds (b) Spores (c) Conidia (d)
Yeast
34. Multicellular fungi are called (a) Molds (b) Spores (c) Conidia (d)
Yeast
35. A saprobe is best described as organisms that (a) Obtain their nutrients from dead or
decomposing organic matter (b) Obtain their nutrients from living and depending on
a viable host (c) Obtain their nutrients from decomposing inorganic matter (d) Lacks
the ability to synthesis their food thus depends on primary and secondary producers for
survival.
36. Fungi are considered important tools of bioremediation because (a) They
posses long and strong network of Hyphae (b) They are found
everywhere in nature (c) Their mycelium are able to
undergo intense bioremediation (d)They are able to produce wide variety of
enzymes
37. Some fungi have proven medically useful because they can be used (a) In
bioremediation (b) In fermentation in brewing industry
(c) In production of certain antibiotic (d) In baking industry
38. Lichens are an association between (a) Fungus and its algae partner (b)
Fungus and bacteria (c) Protozoa and bacteria (d) Fungi and
a photosynthetic algae
39. Which statement best describes Myxomycota (i) They behave
like animals at vegetative stage and behave like fungi at reproductive stage.
(ii) They behave like fungi at vegetative stage and behave like animals at reproductive
stage (iii) The body of some members is known as plasmodium
(iv) They reproduces particularly sexually and not asexually (a)i, iii only (b) ii,
iii only (c) iii only (d) ii, iii, iv
40. The functional equivalent of cholesterol found in cell membranes of fungi and some
protists, as well as, the steroid precursor of vitamin D2 is (a) Lipid (b) Ergosterol
(c) Waxes (d) Phospholipids
41. Simi was diagnosed with a______ disease called athletic foot, the probable causal
organism is_______ (a) Bacterial, Trichophyton rubrum (b) Fungal, Trichophyton
lysin (c) Bacterial, Trichophyton lysin (d) Fungal, Trichophyton rubrum
42. If the study of bacteria is Bacteriology, the study of fungi is (a) Mycology (b)
Ornithology (c) Fungulogy (d) Fungiology
43. Mushrooms are a type of which of the following (a) Conidia (b) Ascus (c)
Polar tubule (d) Basidiocarp
44. Which of the following is the most common cause of human yeast infections? (a)
Candida albicans (b) Blastomyces dermatitidis (c) Cryptococcus neoformans
(d) Aspergillus fumigatus
45. Which of the following is an ascomycete fungus associated with bat droppings that can
cause a respiratory infection if inhaled?
(a) Candida albicans
(b) Histoplasma capsulatum (c) Rhizopus stolonifera (d) Trichophyton
rubrum
Theory
1. Define the following terms (i) Mycorrhiza (ii) Sporangiophore (iii) Lichen
(iv) Spores (v) Gametangium
2. Explain why the study of fungi such as yeast and molds is within the discipline of
Microbiology.
3. Differentiate between the following (i) Sporangiophore and Sporangium (ii)

Plasmogamy and Karyogamy (iii) Mutualism and Saprophytism (iv)
Hyphae and Mycelium (v) Sexual and Asexual reproduction (vi)
Homothallic and Heterothallic fungi (vii) Myxomycota and Eumycota
(viii) Coenocytic and Aceonocytic fungi (ix) Chitin and Ergosterol (x)
Unicellular and Multicellular organism
4. Fungi are more related to animals than plants. Justify this statement.
5. A student travelling by road on her way for a national examination made a stop to use a
public toilet due to the pressure coming from the content of her bladder. Afterwards,
she continued on her journey. Days later, she observed mild itches at her genital region
which she ignored, the itching grew with time to become severe and accompanied by a
foul smelling discharge from her genitals. (a) Predict the
most likely urinogenital disease challenging this student. (b) State the causal
organism of this disease (c)What type of microorganism is
implicated in this disease. (d) Is it a contagious disease? Justify your
answer (e) Enumerate 3 preventive measures to be taken
against this disease
6. Enumerate 10 differences and 5 Similarities between fungi and bacteria.
7. Succinctly explain reproduction in a named fungi
8. State the 5 classes of Eumycota with 5 characteristic each

9. Using diagrams explain the differences between Septate and Non-septate fungi
10. State 5 Economic importance of fungi

11. Fungi are exclusively eukaryotic saprophytes. TRUE or FALSE. (b) Justify your
answer

1. C
2. B
3. D
4. A
5. C
6. D
7. A
8. D
9. A
10. C
11. D
12. A
13. C
14. C
15. A
16. B
17. A
18. B
19. D
20. D
21. B
22. D
23. B
24. A
25. A
26. C
27. B
28. B
29. C
30. A
31. C
32. B
33. D
34. A
35. A
36. D
37. C
38. D
39. A
40. B
41. D
42. A
43. D
44. A
45. B
THEORY
2. Fungi are included within the field of microbiology because of the prevalence of many
unicellular yeasts; also, some spores of macroscopic fungi are microscopic.
3.
4. Fungi are more related to animals than plants because of the following Similarities.
● They are both eukaryotic
● They both store food as glycogen
● They are both heterotrophs
5. (a) Vaginal candidiasis.

(b) Candida albicans
(c) A parasitic fungi
(d) It is proven to be contagious if a healthy person shares underwear and other
personal items with an infected person. But it cannot be transferred from one person to
another through physical contact.
(e) Avoid the use of unkempt public conveniences
Avoid patronising used underwear vendors.
Maintainance of effective personal hygiene always
Avoid sharing underwear with family and friends.
6.
FUNGI BACTERIA
They are eukaryotic They are prokaryotes
They have a cell wall that contains chitin They have a cell wall that contains
peptidoglycan layer.
They are all heterotrophs Some are autotrophs while some are
heterotrophs
They have a nucleus which contains the They have a nucleoid which contains the
genetic material genetic material
They posses 80s ribosome They posses 70s ribosome
They attach to a surface using Rhizoids They attach to a surface using pilli
Some are microscopic while some are They are all microscopic
macroscopic
Some are unicellular while some are They are all unicellular
multicellular
There is presence of cellular organelles There is absence of cellular organelles
They reproduce both sexually and asexually. They reproduce asexually through binary
fission
Similarities
● They both possesses genetic materials
● They both possesses cell wall which confers protection and shape to cell.
● They are both heterotrophs except Cyanobacteria
● They are both parasitic to plants and animals
● They both possesses cytoplasm in their cell which contains dissolved organic substances.
● They both posses ribosomes.
● They both possesses a semi-permeable membrane that regulates passage of substances
in and out of the cell.
● They can both be killed under extremely high temperatures.

10. They are medically useful in the production of penicillin. E.g Penicillium notatum
They are industrially useful in the production of baked product. E.g Saccharomyces
cerevisea
They cause spoillage in food product. E.g Mucor
They are used as biological pesticide on agriculture. E.g Trichoderma sp
They are implicated in several human and animal diseases such as Athletic foot
(Trichophyton rubrum) and Aspergillosis (Aspergillus niger) respectively
The are used in fermentation in brewery industry. E.g Saccharomyces cerevisea
In mycorrhiza, the fungi partner absorbs and supplies the plant with water, minerals
and other essential growth factor thereby promoting the growth and quality of plants.
11. True. Eukaryotic because they have a true nucleus and saprophytic because they feed
and derive energy from dead organic matter.
CHAPTER 5
5.1 STAINING TECHNIQUE
Staining can be defined as a technique used to enhance contrast and visibility in samples,
generally at the microscopic level by the addition of stains and dyes.
In their natural state, most of the cells and microorganisms that we observe under the
microscope lack color and contrast. This makes it difficult, if not impossible, to detect important
cellular structures and their distinguishing characteristics without artificially treating specimens
with stains or dyes.
5.2 BASIC & ACIDIC DYE

Staining is almost always applied to color certain features of a specimen before examining it
under a light microscope. Stains or dyes contain salts made up of a positive ion and a negative
ion. Depending on the type of dye, the positive or the negative ion may be the chromophore
(the colored ion); the other, uncolored ion is called the counterion.
When the chromophore is the positively charged ion, the stain is classified as a basic dye; if the
negative ion is the chromophore, the stain is considered an acidic dye.
Dyes are selected for staining based on the chemical properties of the dye and the specimen
being observed. In most cases, it is preferable to use a positive stain, a dye that will be
absorbed by the cells or organisms being observed, adding color to objects of interest to make
them stand out against the background. However, there are scenarios in which it is
advantageous to use a negative stain, which is absorbed by the background but not by the cells
or organisms in the specimen. Negative staining produces an outline or silhouette of the
organisms against a colorful background
Because cells typically have negatively charged cell walls, the positive dye tends to stick to the
cell walls, making them positive stains.
Thus, commonly used basic dyes includes basic fuchsin, crystal violet, malachite green,
methylene blue, and safranin typically serve as positive stains.
On the other hand, the negatively charged chromophores in acidic dyes are repelled by
negatively charged cell walls, making them negative stains. Commonly used acidic dyes include
acid fuchsin, eosin, and rose bengal.
5.3 SIMPLE & DIFFERENTIAL STAINING
Some staining techniques involve the application of only one dye to the sample; others require
more than one dye.
In simple staining, a single dye is used to emphasize particular structures in the specimen. A
simple stain will generally make all of the organisms in a sample appear to be the same color,
even if the sample contains more than one type of organism.
In contrast, differential staining distinguishes organisms based on their interactions with
multiple stains, two organisms in a differentially stained sample will appear to be different
colors. In other words, differential staining involves the use of more than one dye or stain.
Differential staining techniques commonly used in clinical settings include Gram staining, acid-
fast staining, endospore staining, flagella staining, and capsule staining.
5.4 GRAM STAINING

The Gram stain procedure is a differential staining procedure that involves multiple steps. It was
developed by Danish microbiologist Hans Christian Gram in 1884 as an effective method to
distinguish between bacteria with different types of cell walls, and even today it remains one of
the most frequently used staining techniques.
Gram-staining is a differential staining technique that uses a fresh culture of not more than
24hrs old, primary stain and a secondary counterstain to distinguish between gram-positive and
gram-negative bacteria.
The aim of Gram staining is to identify bacteria cells with thick and thin peptidoglycan layer
within its cell walls.
STEP 1
Crystal violet, a primary stain, is flooded on the heat-fixed smear, giving all of the cells a purple
color, then rinsed briefly with slow running water to remove residual stain.
Effect on Cells: Stains cells purple
STEP 2
Logul iodine, a mordant, is flooded on the smear , then the smear is rinsed again.
A mordant is a substance used to set, stabilize and adhere stains or dyes on the cell wall of
bacteria cell in the smear; in this case, iodine acts like a trapping agent that together with the
crystal violet, forms a crystal violet–iodine complex which stays contained in thick layers of
peptidoglycan in the cell walls.
Effect on Cells: Cell remains purple
STEP 3
Decolorizing agent is added, usually ethanol or an acetone/ethanol solution.
Cells that have thick peptidoglycan layers in their cell walls are much less affected by the
decolorizing agent; they generally retain the crystal violet dye and remain purple. However, the
decolorizing agent more easily washes the dye out of cells with thinner peptidoglycan layers,
making them again colorless.
Effect on Cells: Gram-positive cells remain purple, Gram-negative cells are colorless.
STEP 4
Secondary/counterstain, usually safranin, is added. This stains the decolorized cells pink and is
less noticeable in the cells that still contain the crystal violet dye.
Effect on Cells: Gram-positive cells remain purple. Gram-negative cells appear pink or red.
RESULT
When observed using a microscope, the resulting image reveals the shape and colour of cells.
For a positive result, the cells will retain the purple colour of crystal violet and thus are Gram-
positive.
For a negative result, the cells will retain the red or pink colour of the safranin and thus are
Gram-negative.
However, there are several important considerations in interpreting the results of a Gram
stain.
First, older bacterial cells may have damage to their cell walls that causes them to appear
gram-negative even if the species is gram-positive. Thus, it is best to use fresh bacterial cultures
for Gram staining.
Second, errors such as leaving on decolorizer too long can affect the results. In some cases,
most cells will appear gram-positive while a few appear gram-negative. This suggests damage
to the individual cells or that decolorizer was left on for too long; the cells should still be
classified as gram-positive if they are all the same species rather than a mixed culture.
5.5 IMPORTANCE OF GRAM STAINING

The differing interactions of bacterial cells with dyes and stains aids to identify microscopic
characteristics of cells.
Besides their differing interactions with dyes and decolorizing agents, the chemical differences
between gram-positive and gram-negative cells have other implications with clinical relevance.
For example, Gram staining can help clinicians classify bacterial pathogens in a sample into
categories associated with specific properties. Gram-negative bacteria tend to be more
resistant to certain antibiotics than gram-positive bacteria due to the presence of external
covering called capsule.
5.6 ACID FAST STAINING

Acid-fast staining is another commonly used differential staining technique.
Acid-fast staining is able to differentiate two types of gram-positive cells: those that have waxy
mycolic acids in their cell walls, and those that do not.
It is a procedure used to stain group of organisms that do not readily take up stains due to the
presence of high concentration of waxy mycolic acid in their cell wall.
Two different methods for acid-fast staining are the Ziehl-Neelsen technique and the Kinyoun
technique.
STEP 1
Primary stain, (carbolfuchsin) a reddish dye is flooded on the smear and heated over a boiling
water which facilitates staining. The smear is rinsed briefly under slow running water to
remove residual stain.
Effect on Cells : Stains cell red
STEP 2
Acid alcohol a decolorizer is flooded briefly on the smear to remove carbolfuchsin.
Effect on Cells: Most cells are decolorized while some still retains the carbolfuchsin.
STEP 3
Methylene blue a counter stain/secondary dye is applied which impacts blue colour to cells.
Effect on Cells : Some cells appear blue while others still retains the colour red of the
carbolfuchsin.
RESULT
When observed using a microscope, the resulting image reveals the shape and colour of cells.
For a positive result, the cells will retain the red colour of carbolfuchsin.
For a negative result, the cells will retain the methylene blue colour.
The fundamental difference between the two carbolfuchsin-based methods is whether heat is
used during the primary staining process.
The Ziehl-Neelsen method uses heat to infuse the carbolfuchsin into the acid-fast cells, whereas
the Kinyoun method does not use heat. Both techniques are important diagnostic tools because
a number of specific diseases are caused by acid-fast bacteria (AFB). If AFB are present in a
tissue sample, their red or pink color can be seen clearly against the blue background of the
surrounding tissue cells
5.7 IMPORTANCE OF ACID FAST STAINING

Acid fast staining is an important diagnostic tool for clinical purposes in identifying a number of
specific disease causing microbes (pathogens) such as those that causes tuberculosis
(Mycobacterium tuberculosis) and leprosy (Mycobacterium leprae) from body sample such as
blood, urine, sputum etc.
5.8 CAPSULE STAINING

Certain bacteria and yeasts have a protective outer structure called a capsule. Since the
presence of a capsule is directly related to a microbe’s virulence (its ability to cause disease),
the ability to determine whether cells in a sample have capsules is an important diagnostic tool.
Capsules do not absorb most basic dyes; therefore, a negative staining technique (staining
around the cells) is typically used for capsule staining. The dye stains the background but does
not penetrate the capsules, which appear like halos around the borders of the cell. The
specimen does not need to be heat-fixed prior to negative staining.
The common negative staining technique for identifying encapsulated yeast and bacteria is
STEP 1
A smear of bacteria cells on a glass slide is formed.
STEP 2
Few drops of India ink or nigrosin is added to the smear.
RESULT
When observed using a microscope, the resulting image reveals the added stain coloured
background but not the capsule leaving halo around each cell shape.
Other capsular stains can also be used to negatively stain encapsulated cells. Alternatively,
positive and negative staining techniques can be combined to visualize capsules: The positive
stain colors the body of the cell, and the negative stain colors the background but not the
capsule, leaving halo around each cell.
5.9 ENDOSPORE STAINING
Endospores are structures produced within certain bacterial cells that allow them to survive
harsh conditions.
Gram staining alone cannot be used to visualize endospores, which appear clear when Gram-
stained cells are viewed. Endospore staining uses two stains to differentiate endospores from
the rest of the cell.
The Schaeffer-Fulton method is the most commonly used endospore-staining technique.
STEP 1
Primary stain, (Malachite green ) a greenish dye is flooded on the smear and heated over a
boiling water which facilitates staining. The smear is rinsed briefly under slow running water to
remove residual stain.
Effect on Cells : Stains cell green
STEP 2
Counterstain/Secondary dye (safranin) is flooded on the smear
RESULT
When observed using a microscope, the resulting image reveals the shape and location of
endospores, if they are present.
For a positive result, the endospores will appear green either within the pink vegetative cells
or completely green, indicating the presence of endospore.
For a negative result, only the pink vegetative cells will be visible indicating that no endospores
are present.
5.10 IMPORTANCE OF ENDOSPORE STAINING

Endospore-staining techniques are important for identifying Bacillus and Clostridium sp, two
genera of endospore-producing bacteria that contain clinically significant species. Among
others, B. anthracis (which causes anthrax) has been of particular interest because of concern
that its spores could be used as a bioterrorism agent. C. difficile is a particularly important
species responsible for the typically hospital-acquired infection known as “C. diff.”
5.11 FLAGELLA STAINING

Flagella (singular: flagellum) are tail-like cellular structures used for locomotion by some
bacteria, archaea, and eukaryotes. Because they are so thin, flagella typically cannot be seen
under a light microscope without a specialized flagella staining technique. Flagella staining
thickens the flagella by first applying mordant (generally tannic acid, but sometimes potassium
alum), which coats the flagella; then the specimen is stained with pararosaniline (most
commonly) or basic fuchsin.
Though flagella staining is uncommon in clinical settings, the technique is commonly used by
microbiologists, since the location and number of flagella can be useful in classifying and
identifying bacteria in a sample. When using this technique, it is important to handle the
specimen with great care; flagella are delicate structures that can easily be damaged or pulled
off, compromising attempts to accurately locate and count the number of flagella

Multiple Choice
1. In differential staining (a) Cells interacts with multiple stains (b) Cells
interacts with biochemical dyes (c) Cells interacts with a single stain (d) Cells
interact with each other to produce microscopic biochemical characteristics.
2. For a positive result of Gram staining, the cells will appear (a) Purple (b)
Green (c) Red (d) blue
3. For a negative result of Gram staining, the cells will appear (a) Purple (b)
4. Which of the following statement is correct (a) Flagella can be seen using a light
microscope (b) Flagella can be seen using an electron microscope (c) Flagella
cannot be seen with light microscope without a specialized flagella staining technique.
(d) Flagella cannot be seen with electron microscope without a specialized flagella
staining technique.
5. Bacillus sp. can be identified using (a) Gram-staining (b) Endospore staining
(c) Acid-fast staining (d) Flagella staining.
6. A dye was discovered to contain positively charged ion. This dye is suspected to be (a)
Eosin (b) saffaranin (c) Malachite blue (d) Malachite green
7. For a negative result of an endospore stain, the endospores will appear (a) Pink
(b) Green (c) Red (d) blue
8. In simple staining (a) Cells interacts with multiple stains (b) Cells
interacts with biochemical dyes (c) Cells interacts with a single stain (d) Cells
interact with each other to produce microscopic biochemical characteristics.
9. A dye was discovered to contain negatively charged ion. This dye is suspected to be
(a) Eosin (b) saffaranin (c) Malachite green (d) Methylene blue
10. Endospore-staining is an example of (a) Differential staining (b) Simple staining
(c) Negative staining (d) None of the above
11. For a positive result of an endospore stain, the endospores will appear (a) Pink (b)
12. A positively charged ion are present in a dye. This dye is considered to be (a) Basic
(b) Neutral (c) Acidic (d) None of the above.
13. A negatively charged ion are present in a dye. This dye is considered to be (a) Basic
(b) Neutral (c) Acidic (d) None of the above.
14. The counter stain in Gram staining is (a) Acid alcohol (b) Safranin (c)
Crystal violet (d)Iodine
15. The secondary stain in Gram staining is (a) Acid alcohol (b) Safranin (c)
16. Endospores are structures in a bacteria cell that (a) Ensures bouyancy of the cell
(b) Confers shape and rigidity to the cell (c) Facilitates adherence of cell to surfaces.
(d) Protects cell from harsh environmental conditions
17. The cell walls of bacteria typically are (a) Positively charged (b) Negatively charged
(c) Neutrally charged (d) None of the above.
18. A mordant is used in (a) Fixing the cell smear to the glass slide (b) Fixing
the counter stain to the cell wall of bacteria (c) Fixing the primary stain to the
cell wall of bacteria. (d) Forms a biochemical complex that enables the bacteria cell
to be easily identifiable.
19. For a positive result of Acid-fast staining, the cells will appear (a) Purple (b)
20. For a negative result of Acid-fast staining, the cells will appear (a) Purple (b)
21. Which of the following statement is correct when a positively charged ion dye is used in
staining (a) The dye will be absorbed by the background but not the cells
forming halos around the cells. (b) The dye will be absorbed by the cells
but not the background. (c) The dye will be visible at the background (d)
The dye will not be visible at the background.
22. Which of the following statement is correct when a negatively charged ion dye is used in
staining (a) The dye will be absorbed by the background but not the cells
forming halos around the cells. (b) The dye will be absorbed by the cells
but not the background. (c) The dye will be visible at the background (d)
The dye will not be visible at the background.
23. It is true that acid-fast bacteria contains concentrated Lipid content in their cell wall
called (a) Peptidoglycan (b) Mycolic acid (c) Teichoic acid (d) Cellulose
24. Clostridium difficile can be identified using (a) Gram-staining (b)
Endospore staining (c) Acid-fast staining (d) Flagella staining.
25. The most commonly used endospore-staining technique is (a) Ziehl-Neelsen
method (b) Kinyoun method (c) Kelhisen method
(c) Schaeffer-Fulton method
26. The mordant in Gram staining is (a) Acid alcohol (b) Safranin (c) Crystal
violet (d)Iodine
27. The primary stain in Gram staining is (a) Acid alcohol (b) Safranin (c)
28. The decolorizing agent in Gram staining is (a) Acid alcohol (b) Alcohol (c)
Acid acetone (d)Acetone solution
29. Acid fast staining is a procedure used to stain group of organisms that do not readily
take up stains due to the presence of high concentration of (a) Peptidoglycan (b)
Mycolic acid (c) Teichoic acid (d) Cellulose
30. Microbes virulence is directly associated to (a) Presence of genetic material (b)
Presence of Cell wall (c) Presence of capsule (d) Presence of peptidoglycan layer
31. Which of the following is the correct pair of non heat fixing staining techniques. (a)
Gram & Capsule Staining (b) Gram & Acid-fast Staining (c) Acid-fast & Endospore
Staining (d) Acid-fast & Capsule Staining
32. Which of the following is the correct pair of heat fixing staining techniques. (a) Gram &
Capsule Staining (b) Gram & Acid-fast Staining (c) Acid-fast &
Endospore Staining (d) Acid-fast & Capsule Staining
33. The best cell culture to be used in Gram staining is (a) 24hrs old (b) 48hrs old
(c) 50hrs old (d) 72hrs old
34. The basic aim of Gram staining is (a) To identify cellular organelles present in a
bacteria cell (b) To differentiate bacteria cell from yeast and molds using
their chemical microscopic properties (c) To identify cells with thick
and thin peptidoglycan layer. (d) To differentiate between prokaryotic
and eukaryotic cells using their chemical microscopic properties.
35. All except one are correct technique in staining (a) Proper disinfection of work
desk (b) The use of old culture (c) Application of decolorizer for a long
period of time (d) Slide rinsing omission.
36. The basic aim of Gram staining is (a) To identify bacteria cells with mycolic
acids in their cell walls (b) To differentiate bacteria cell from yeast and molds
using their chemical microscopic properties (c) To identify cells with
thick and thin peptidoglycan layer. (d) To differentiate between
prokaryotic and eukaryotic cells using their chemical microscopic properties
37. The basic aim of acid-fast staining is (a) To identify bacteria cells with mycolic
acids in their cell walls (b) To differentiate bacteria cell from yeast and molds
using their chemical microscopic properties (c) To identify cells with
thick and thin peptidoglycan layer. (d) To differentiate between
prokaryotic and eukaryotic cells using their chemical microscopic properties
38. The spores of Bacillus anthracis can be used in (a) Bioremediation (b)
Bioconservation (c) Bio-degradation (d) Bio-
terrorism
39. Tail-like structure used in locomotion in bacteria is (a) Pilli (b) Flagella (c)
capsule (d) Hyphae
40. Gram negative bacteria are more resistant to Gram positive because of the presence of
(a) Pilli (b) Flagella (c) Capsule (d) Hyphae
41. Which scientist developed Gram staining (a) Christian Gram (b) Robert
Gram (c) Joseph Gram (d) Hangar Gram
42. Process whereby chemical substances are added to cells to make it microscopically
visible is (a) Staining (b) Incubation (c)Tyndallization
(d)Dyeing
THEORY
1. Define the following (i) Staining (ii) Smear (iii) Mordant (iv)
Decolorizer (v) Virulence
2. Compare the staining technique of Gram and Acid fast Staining.
3. Compare the staining technique of Acid fast and Endospore staining.
4. Differentiate between (i) Differential & Simple Staining (ii) Basic

& Acid dye (iii) Bioterrorism and Bioremediation
5. Appropriately fill in the table below
PRIMARY SECONDARY DECOLORIZ MORDANT COUNTER

DYE DYE ER STAIN
ACID-FAST
STAINING
GRAM
STAINING
ENDOSPOR
E STAINING
6. Differentiate between Ziehl-Neelsen and Kinyoun technique used in Acid-fast staining
7. Enumerate 3 significance of Gram staining
8. Enumerate 3 limitations of Gram staining
9. Cell cultures of 48hrs and above old is best for Gram staining. TRUE or FALSE (b)
Justify your answer.
10. Compare the action of positive ions and negative ions in staining technique.
11. State 5 errors to be avoided when carrying out Gram staining.
12. Compare and contrast between Gram-positive and Gram-negative bacteria.

1. A
2. A
3. C
4. C
5. B
6. D
7. A
8. C
9. A
10. A
11. B
12. A
13. C
14. B
15. B
16. D
17. B
18. C
19. C
20. D
21. B
22. A
23. B
24. B
25. A
26. D
27. C
28. B
29. B
30. C
31. A
32. C
33. A
34. C
35. B
36. C
37. A
38. D
39. B
40. C
41. A
42. A
THEORY
1. Staining can be defined as a technique used to enhance contrast and visibility in
samples, generally at the microscopic level by the addition of stains and dyes.
Smear is a sample of tissue, cells or other material taken from part of the body, spread
thinly on a microscope glass slide by a loop or swab for microscopic examination,
typically for medical diagnosis.
A mordant is a substance used to set, stabilize and adhere stains or dyes on the cell
wall of bacteria cell in the smear.
A decolorizer is a chemical agent that used in staining that facilitates the washing of dye
out of cells with thinner peptidoglycan layers, making them again colorless.
Virulence is defined as the degree of pathology caused by a pathogenic organism.

4. (i) Make reference to Chapter 5.3 (ii) Make reference to Chapter 5.2
(iii)
Bioterrorism Bioremediation
The use of macro or microbial agents or The use of mainly microorganisms, plants,
other harmful biological or biochemical or enzymes to detoxify, degrade, and
substances as weapons of terrorism. eliminate hazardous pollutants and
contaminants from the soil and other
environment.
5.
PRIMARY SECONDARY DECOLORIZ MORDANT COUNTER

DYE DYE ER STAIN
ACID-FAST CARBOLFUC METHYLENE ACID ABSENT METHYLENE

STAINING HSIN BLUE ALCOHOL BLUE
GRAM CRYSTAL SAFRANIN ALCOHOL LOGUL SAFRANIN

STAINING VIOLET IODINE
ENDOSPOR MALACHITE SAFRANIN ABSENT ABSENT SAFRANIN

E STAINING GREEN
7. (i) It helps identify bacteria cells with thick and thin peptidoglycan layer
(ii) It helps in enhancing microscopic visuality of microorganisms.
(iii) It proves to be useful in clinical diagnosis and treatment of various pathogenic
microorganisms and their diseases.
(iv) It allows for various methods of testing either from sputum or from blood samples.
(v) It is a simple and cost-effective procedure.
( vi) It gives quick result when examining infections to enable quick medical treatment.
8. (i) Inability to visualize atypical bacteria (Atypical bacteria are bacteria that do not color
with gram-staining but rather remain colorless: they are neither Gram-positive nor
Gram-negative).
(ii) Inability to establish preliminary substantial morphologic identification when used in
environmental micrbiology.
(iii) It can be easily misinterpreted thus it is used in combination with other molecular
and traditional techniques in identifying bacteria.
9. False, because older bacterial cells may have damage to their cell walls that causes them
to appear gram-negative even if the species is gram-positive. Thus, it is best to use fresh
bacterial cultures for Gram staining.
10. Dye with positive ions will be absorbed by the cells or organisms being observed,
adding color to organelles of interest to make them stand out and visible against the
background.
While, dye with negative ions, is absorbed by the background but not by the cells or
organisms in the specimen. It produces an outline or silhouette of the organisms against
a colorful background
11. (i) Avoid using an old cell culture in Gram staining

(ii) Avoid working on a contaminated work desk and site.
(iii) Avoid leaving the decolorizer on the smear for a long time
(iv) Avoid omitting rinsing steps in Gram staining
(v) Avoid using non-sterilized apparatus and material for Gram staining.
12.
GRAM POSITIVE GRAM NEGATIVE
COLOUR PURPLE REDDISH PINK
PEPTIDOGLYCAN LAYER THICK THIN
TEICHOIC ACID PRESENT ABSENT
CAPSULE ABSENT PRESENT
ENDO-TOXIN ABSENT PRESENT
PORIN PROTEIN ABSENT PRESENT
PERIPLASM ABSENT PRESENT
SENSITIVITY TO PENICILLIN MORE SUSCEPTIBLE LESS SUSCEPTIBLE
SENSITIVITY TO LYSOZYME MORE SUSCEPTIBLE LESS SUSCEPTIBLE

FORM WHEN FORMS PROTOPLAST FORMS SPHEROPLAST
PEPTIDOGLYCAN IS
REMOVED
CHAPTER 6
6.1 MICROBIAL GROWTH PHASES

Microbial growth is defined as irreversible increase in number of cell, size, mass, or colony
diameter of a population of microbial cell.
There are 6 basic growth requirement of microorganisms and they are
(i) Carbon source (ii) Energy source (iii) Nitrogen source (iv) Water
(v) Growth factor (vi) Mineral elements
Microorganisms utilitises these materials to increase in size. The time it takes for a population
to double in number is called Generation time or Doubling time.
6.2 BATCHED & CONTINUOUS SYSTEM

In batched system, microorganisms are grown in a broth contained in a tube or flask or in an
agar plate where nutrients are not renewed nor are waste product removed. Here the growth
phases includes Lag, Log/Exponential, Stationary, Death, Prolonged death phases.
Under this condition, cell population increases predictably and eventually declines. As the
population grows, it follows a pattern called growth curve
However in contrast, continuous system is done in an open system through a chemostat and
can only maximally reach exponential /log phase because nutrients are continually added and
waste product are continually removed.
6.3 THE GROWTH CURVE
LAG PHASE
● After Inoculation and incubation, number of cells do not increase immediately, they go
through lag phase or tooling up phase.
● Here, bacteria adapt themselves to growth and environmental conditions such as
oxygen, tension and temperature.
● It is the period where the individual bacteria are maturing and not yet able to divide.
● During the lag phase of the bacterial growth cycle, synthesis of RNA, enzymes,
ribosomes and other macromolecules are synthesised.
● During the lag phase cells change very little because the cells do not immediately
reproduce in a new medium. This period of little to no cell division is called the lag phase
and can last for 1 hour to several days.
● During this phase cells are not dormant.
LOG/EXPONENTIAL PHASE
● Here, cells divides at a constant rate and their number increases by the same
percentage during each time interval.
● Here, the generation time is measured during the period of active multiplication.
● At the initial phase of exponential growth, all cell activities are directed towards
increasing the cell mass.
● It is a period characterized by cell doubling. The number of new bacteria appearing per
unit time is proportional to the present population.
● If growth is not limited, doubling will continue at a constant rate so both the number of
cells and the rate of population increase doubles with each consecutive time period.
● There's rapid division of cell at constant rate.
● There's rapid utilisation of nutrients.
● Cells possesses uniform physiological characteristics.
● This period is also known as trophophase in commercial fermentation.
● Compounds such as amino acids and nucleotides are produced.
● The actual rate of this growth phase depends upon the growth conditions, which affect
the frequency of cell division events and the probability of both daughter cells surviving.
● Under controlled conditions, cyanobacteria can double their population four times a day
and then they can triple their population.
● Exponential growth cannot continue indefinitely because the medium is soon depleted
of nutrients and enriched with wastes.
STATIONARY PHASE
● The stationary phase is often due to exhaustion of essential nutrients, and/or the
formation of an inhibitory product such as an organic acid.
● The total number of viable cells in the overall population remains relatively constant.
● But actually, the main activity going on is that some cells are dying while others are
multiplying.
● The dead cells release peptide and nucleic acid which serves as source of nutrient and
energy to fuel the growth of other cells.
● Secondary metabolites such as antibiotic is synthesised.
● Here, death rate equals reproduction rate.
● Spore formers begin to sporulate
● There is accumulation of toxic products and materials.
● The number of new cells created is limited by the growth factor and as a result the rate
of cell growth matches the rate of cell death.
● The result is a “smooth,” horizontal linear part of the curve during the stationary phase.
● Mutations can occur during stationary phase.
DEATH PHASE
● Depletion of nutrients is very rapid
● Rapid accumulation of toxic materials
● Here , bacteria begin to die rapidly.
● This could be caused by lack of nutrients, environmental temperature above or below
the tolerance band for the species, or other injurious conditions.
● Rapid sporulation by Spore formers
● Period when the total number of viable cell in the population decreases.
PHASE OF PROLONGED DECLINE

● It is marked by a very rapid decrease in number of viable cell in the population.
● Extensive accumulation of waste product
● It could last from days to years.
● The dynamic system generates a successive wave of slightly modified population each
more fit to survive than the previous one thus survival of the fittest.
6.4 ASEPTIC & SAFETY PRECAUTIONS IN THE LABORATORY
● Ensure disinfection of work bench with a potent disinfectant.

● Ensure proper Sterilisation of laboratory apparatus and materials before use.
● Ensure all solutions to be used are sterilised in the autoclave at 121oC.
● Ensure the presence of spirit lamp close to work site.
● Ensure the proper use of laboratory coat and other personal protective equipment.
● Ensure to sterilize inoculation loop before and after each use.
● Ensure talking and eating is totally avoided when working.
● Ensure total and complete covering of hair drops to prevent cross contamination.
● In case of sneezing or coughing, ensure such is done using a handkerchief or a bent
elbow to prevent cross contamination.
● Ensure all electrical equipment are disconnected from power source when not in use.
● Ensure all wraps, bottles and waste are properly disposed in the waste bin.
● Ensure all slippery items or substances are eliminated from the floor to prevent an
accident.
6.5 IMMUNITY
It is simply defined as a state of non-susceptibility of an organism to any disease causing agent.
It is the capacity to recognise, analyse, provoke immune response and eliminate intruding and
invading foreign bodies (antigen) from the body with great effectiveness and speed.
Immunity can be of two types.

● Natural immunity
● Artificial immunity
6.6 NATURAL IMMUNITY
This is a type of immunity generated, synthesised and imposed by the body without any
external assistance and aid.
It is further subdivided into two

● Naturally Acquired Active Immunity (NAAI)
● Naturally Acquired Passive Immunity (NAPI)
NAAI- This is a type of immunity autonomously generated by the body due to the action of
memory cells produced during previous infection.
When the body is exposed to an infection it provokes immune response eliciting antibody
production to combat the infection producing memory cells in the process. This memory cell
ensures fast and lasting action against any of such invading antigen next time.
NAPI- This is immunity transferred from one person to another through natural means such as
in prenatal and postnatal relationships between mother and child.
Naturally Acquired Passive Immunity occurs during pregnancy, in which certain antibodies are
passed from the maternal blood into the fetal bloodstream
Here, no memory cells are produced.
6.7 ARTIFICIAL IMMUNITY
Artificial immunity is a means by which the body is given immunity against a disease by
intentional exposure to small quantities of that infectious entity.
It is further subdivided into two

● Artificially Acquired Active Immunity (AAAI)
● Artificially Acquired Passive Immunity (AAPI)
AAAI- This is immunity produced by intentional exposure of a healthy person to antigens
contained in a vaccine, so as to provoke lasting immune response and elicit antibody
production to safe-guard the body against any of such pathogen contained in the vaccine.
Here, memory cells are produced.
AAPI- Artificially-acquired passive immunity is an immediate, but short-term immunity provided

by the injection of antibodies, such as gamma globulin, that are not produced by the recipient's
cells.
These antibodies are developed in another individual or animal and then injected into another
individual to confer immunity against a particular disease e.g. Tetanus
Here, no memory cells are produced thus the immunity is for a short term.

Multiple Choice
1. The microbial growth curve is associated with (a) Batch System (b) Botched system.
(c) Continuous system (d) Open system
2. In lag phase of a microbial growth curve. (a) Secondary metabolites are produced
(b)Bacteria begins to adapt to growth and environmental conditions. (c) There is
rapid utilization of nutrients (d) There's rapid sporulation by spore formers.
3. Secondary metabolites such as antibiotic is synthesised at (a)Lag phase (b)Log phase
(c) Stationary phase (d)Death phase
4. Bacteria adaptation to growth and environmental conditions occurs at (a)Lag phase
(b)Log phase (c) Stationary phase (d)Death phase
5. Sterilization using an autoclave is done at. (a)111oC. (b)121oC. (c)131oC.
o
(d)141 C
6. Rapid utilization of nutrients is a characteristic of (a)Lag phase (b)Log phase (c)
Stationary phase (d)Death phase
7. Rapid depletion of nutrients is a characteristic of (a)Lag phase (b)Log phase (c)
Stationary phase (d)Death phase
8. Rapid sporulation by spore formers is a characteristic of (a)Lag phase (b)Log phase

(c) Stationary phase (d)Death phase
9. Memory cells are produced during (i) Naturally acquired Active Immunity. (i) Artificially
acquired Active Immunity. (iii)Naturally acquired passive Immunity. (iv) Artificially
acquired passive Immunity. (a)i & iii. (b)ii &iv. (c)i & ii. (d) iii &iv
10. Immunity can be defined as. (a) State of susceptibility of an organism to disease.
(b) State of non susceptibility of an organism to death (c) State of non
susceptibility of an organism to disease. (d) State of susceptibility of an organism to
death
11. Period when the total number of viable cell in the population decreases is (a)Lag phase
(b)Log phase (c) Stationary phase (d)Death phase
12. Successive wave of slightly modified population each more fit to survive than the
previous one is a characteristic of. (a)Lag phase (b)Log phase (c) Stationary
phase (d) Death phase
13. The number of new cells created is limited by the growth factor and as a result the rate
of cell growth matches the rate of cell death; this is. (a)Lag phase (b)Log phase
(c) Stationary phase (d) Death phase
14. Which of the following is not a basic growth requirement of microorganisms (a)
Carbon source (b)Flourine source (c) Nitrogen source (d) Water
15. Irreversible increase in number of cell, size, mass, or colony diameter of a population of
microbial cell is (a) Microbial development (b) Microbial growth.
© Microbial Growth curve. (d) Microbial Growth rate
16. Which of the following is not an aseptic technique in the laboratory (a)Disinfection of
work bench with a potent disinfectant.
(b) Sterilisation of laboratory apparatus and materials before use.
(c)Presence of spirit lamp close to work site.
(d)Use of laboratory coat and other personal protective equipment.
17. Which is the following is not a safety technique in the laboratory

(a) Presence of spirit lamp close to work site
(b)Use of laboratory coat and other personal protective equipment
(c) Disconnection of all electrical equipment from power source when not in use.
(d)Elimination of slippery items or substances from the floor
18. Rapid division of cell at constant rate is common in (a)Lag phase (b)Log phase
19. The release of peptide and nucleic acid from dead cells which serves as source of
nutrient and energy to fuel the growth of other cells is typical in (a)Lag phase
(b)Log phase (c) Stationary phase (d) Death phase
20. Cells possesses uniform physiological characteristics. (a)Lag phase (b)Log phase
21. Trophophase also refers to (a)Lag phase (b)Log phase (c) Stationary phase
(d) Death phase
22. The smooth horizontal linear part of the growth curve represents. (a)Lag phase
(b)Log phase (c) Stationary phase (d) Death phase
23. The smooth ascending part of the growth curve represents. (a)Lag phase (b)Log phase
24. The smooth descending part of the growth curve represents. (a)Lag phase (b)Log
phase (c) Stationary phase (d) Death phase
25. Short term Immunity is covered by (a) Passive Immunization (b) Active
Immunization (c) Natural Immunization. (d) Artificial Immunization
26. The time it takes for a population to double in number is (a) Doubling time
(b)Generating time. (c) Maturity time. (d) None of the above
27. A type of immunity autonomously generated by the body due to the activities of
previous infection is (a) Naturally acquired Active Immunity. (b) Artificially acquired
Active Immunity. (c)Naturally acquired passive Immunity. (d) Artificially acquired
passive Immunity.
28. Nutrients are continually added and waste product are continually removed in (a)
Batch System (b) Botched system. (c) Continuous system (d) Closed system
29. Nutrients are continually depleted and waste product are continously removed in (a)
Batch System (b) Botched system. (c) Continuous system (d) Open system
30. In a continuous system, which of the following is not present. (i)Lag phase (ii)Log
phase (iii) Stationary phase (iv) Death phase (a)i & ii. (b)ii & iii. (c)iii & iv.
(d)i & iv
31. In a continuous system, which of the following is present. (i)Lag phase (ii)Log phase
(iii) Stationary phase (iv) Death phase (a)i & ii. (b)ii & iii. (c)iii & iv. (d)i & iv
32. A chemostat is used in (a) Batch System (b) Botched system. (c) Continuous system
(d) Closed system
33. A chemostat is not used in (a) Batch System (b) Botched system. (c) Continuous
system (d) open system
34. Tooling up also refers to(a)Lag phase (b)Log phase (c) Stationary
phase (d) Death phase
35. A type of immunity produced by intentional exposure of a healthy person to antigens
contained in a vaccine is (a) Naturally acquired Active Immunity. (b) Artificially acquired
Active Immunity. (c)Naturally acquired passive Immunity. (d) Artificially acquired
passive Immunity.
36. Irreversible increase in number of cell, size, mass, or diameter of an organism is
(a) Microbial development (b) Growth. © Microbial Growth
(d) Microbial Growth rate
37. Rapid accumulation of toxic materials is a characteristic of (a)Lag phase (b)Log phase
38. Antibodies are developed in another individual or animal and then injected into another
individual to confer immunity in (a) Naturally acquired Active Immunity. (b) Artificially
acquired Active Immunity. (c)Naturally acquired passive Immunity. (d) Artificially
acquired passive Immunity.
39. A type of Immunity confered on the foetus through the bloodstream of the mother is
(a) Naturally acquired Active Immunity. (b) Artificially acquired Active Immunity.
(c)Naturally acquired passive Immunity. (d) Artificially acquired passive Immunity.
40. Accumulation of toxic products and materials occurs at (a)Lag phase (b)Log phase
41. After Inoculation and incubation, number of cells do not increase immediately, they go
through (a)Lag phase (b)Log phase (c) Stationary phase (d)
Death phase
THEORY
1. Define the following
(I) Immunity
(ii)Antibody
(iii) Vaccine
(iv) Antigen
(v) Microbial Growth
2. State 3 differences and similarities each between Batch and Continuous system
3. State 2 differences each between the following pair
(I) Naturally acquired active Immunity & Naturally acquired passive Immunity
(ii) Artificially acquired active Immunity & Artificially acquired passive Immunity
4. State 4. characteristic feature of the following microbial phases

(I) Lag phase
(ii)Log phase
(iii) Stationary phase
(iv) Death phase
5. Draw a Microbial Growth curve, on it identify,

(I)Lag,log, stationary,death,and prolonged death phases
(ii) The phase with antibiotic production
(iii) Cell adaptation Stage
(iv) Rapid sporulation stage
(v) Rapid growth stage
(vi) Survival of the fittest phase
(vii) Decrease in viable cell phase
(viii) Production of Amino acid and nucleotide stage
(ix) Stage with uniform physiological characteristic of cell.
(x) RNA, enzymes and ribosome production stage.
6. Differentiate between Aseptic and Safety precaution in the laboratory using relevant
examples.
7. State 6 Basic Growth requirement of microorganisms.

1. A
2. B
3. C
4. A
5. B
6. B
7. D
8. D
9. C
10. C
11. D
12. D
13. C
14. B
15. B
16. D
17. A
18. B
19. C
20. B
21. B
22. C
23. B
24. D
25. A
26. A
27. A
28. C
29. A
30. C
31. A
32. C
33. A
34. A
35. B
36. B
37. D
38. D
39. C
40. C
41. A
THEORY
1. Immunity is defined as a state of non-susceptibility of an organism to any disease
causing agent.
Antibody is a chemical substance produced by the body to combat infectious agents as
a result of response to an antigenic stimulus.
Vaccine is a biological substance constituted of weakened or killed form of microbes,

its toxins or its surface protein, that when introduced into a living host, stimulates
production of antibodies and induces immunity against one or several diseases.
Antigen – A substance of internal chemical complexity that is easily solubilized such

that when administered, provokes immune response, eliciting antibody production
which confers lasting immunity to the host.
Microbial growth is defined as irreversible increase in number of cell, size, mass, or
colony diameter of a population of microbial cell.
2.
BATCH SYSTEM CONTINUOUS SYSTEM

The microbial growth phases includes Lag, The microbial growth phases includes Lag
Log, Stationary, Death and Phase of & Log phases.
prolonged death.
Nutrient is continously depleted without Nutrient is replenished continously

replenishment
The use of a chemostat is not necessary. The use of a chemostat is significant and
necessary
Microbial waste product is accumulated Microbial waste product is removed and

continously eliminated continously.
SIMILARITIES
● They are both used to culture desirable microorganisms.
● Waste product by microorganisms are produced in both systems.
● They both significantly display Lag and Log phases of microbial growth.
● They both require optimum temp for growth of cells to occur.
3.
NAAI NAPI
It is a type of immunity generated by the It is a type of immunity transferred from

body due to the activities of previous one person to another through natural
infection means such as in prenatal and postnatal
relationships between mother and child.
Memory cells are usually produced Memory cells are not produced
Immunity is long lasting Immunity is short lived

AAAI AAPI
It is a type of immunity produced by It is a type of immunity provided by the

intentional exposure of a healthy person extraction and injection of antibodies
to antigens contained in a vaccine. developed in a healthy individual or
animal into a susceptible individual to
confer immunity against a particular
disease
Memory cells are usually produced. Memory cells are not produced.
Immunity is long lasting Immunity is short lived.

6.
ASEPTIC TECHNIQUE SAFETY PRECAUTION
This is a set of rules observed in the These are steps necessary to prevent
laboratory to prevent cross occrence of accident either by fire,
contamination and infection. chemicals etc.
It includes the disinfection of work bench It includes the proper use of laboratory
with a potent disinfectant. coat and other personal protective
equipment.
It also includes sterilisation of laboratory

apparatus and materials before use.
CHAPTER 7
7.1 MICROBIAL ECOLOGY
Ecology is the scientific study of the relationship and interaction between living organisms and
their physical environment of matter and energy.
Microbial ecology is the study of the relationship of microorganisms with each other and their
environment.
Microbial Ecosystem is a self sustaining unit comprising of biotic (microorganisms) and abiotic
factors (sunlight, temperature, rainfall, salinity, pH, humidity etc) interacting with each other.
Biosphere is the region of the earth inhabited by microorganisms. It's also called ecosphere
Lithosphere is the solid part of the earth crust e.g. Rocks, mountain etc.
Hydrosphere is the aquatic part of the ecosphere accounting for ⅔ of the earth crust
Atmosphere is the gaseous part of the biosphere .
Microbial ecological niche is a role or position a microorganism is adapted to in an ecosystem.
Micro-environment is all external factors surrounding, affecting and influencing a particular

microorganism.
7.2 MICROBIAL NUTRIENT ACQUISITION
Microorganisms can be categorised according to their trophic and food source.
Basically 3 trophic levels exist

(i) Primary producer
(ii) Consumers
(iii) Decomposers
PRIMARY PRODUCERS
They are basically autotrophs, they convert carbondioxide (CO2) into organic materials.
Producers includes photoautotrophs which use sunlight for energy Example includes
Cyanobacteria
chemolithoautotrophs which oxidises inorganic compounds (sulfur, nitrogen, iron, ammonia

etc) to derive energy.
Examples include Nitrosomonas, Nitrobacteria etc.
They are found in deep sea hydrothermal vents, and often live in symbiotic relationships with
other invertebrates.
Primary producers generally serve as sources of food for consumers.
CONSUMERS
They are basically heterotrophs because they utilitise organic materials and rely on the
activities of primary producers. Examples include fungi, protozoa and some bacteria.
DECOMPOSERS
These are heterotrophs that digest the dead remains of primary producers and consumers. The
fresh or partially decomposed organic matter used as food sources includes carcasses, feaces
and plant litters.
They specialise in digesting complex materials like Cellulose converting them into smaller
molecules that can be more readily reused by other microorganisms.
The complete breakdown of organic molecules into inorganic molecules like ammonia,
sulphate, phosphate is called mineralization
Example includes All fungi.
7.3 MICROBIAL INTERACTIONS

This refers to the inter and intra relationship between various microorganisms.
Ectosymbiosis is a relationship that exist between a smaller organism located on the surface of
a larger organism. The smaller organism here is referred to as ectobiont Example:
Dermatophytes on the skin of man.
Endosymbiosis is a relationship that exist between a smaller organism that exists inside a larger
organism. The smaller organism here is referred to as endobiont. Example: Bacteria cells in the
gut of human.
TYPES OF INTERACTIONS
Microbial interaction may be positive such as mutualism, proto-cooperation, commensalism or

may be negative such as parasitism, predation or competition
POSITIVE INTERACTIONS
(1) MUTUALISM :
● It is defined as a relationship in which each organism in interaction benefits from the

relationship or association.
● It is an obligatory relationship in which mutualist and host are metabolically dependent

on each other.
● Mutualistic relationship is very specific where one member of association cannot be

replaced by another species.
● Mutualism require close physical contact between interacting organisms.
● Relationship of mutualism allows organisms to exist in habitat that could not be

occupied by either species alone.
● Mutualistic relationship between organisms allows them to act as a single organism.
Examples of mutualism:
(i) Lichens:
● Lichens are excellent example of mutualism.
● They are the association of specific fungi and certain photosynthetic genus of algae.
● In lichen, fungal partner is called mycobiont and algal partner is called phycobiont
which is a member of cycanobacteria and green algae (Trabauxua).
● Because phycobionts are photoautotrophs, the fungus get its organic carbon directly
from algal partner, in turn fungi protects the phycobiont from extreme conditions and
also provide water and minerals to algae.
● Lichen grow very slowly but are able to colonies habitat that do not permit the growth
of other organisms.
● Most lichens are resistant to high temperature and drying.
(ii) Protozoan-termite:
● Protozoan-termite relationship is the classical example of mutualism in which flagellated
protozoan lives in the gut of termites.
● These flagellated protozoan feeds on diet of carbohydrates acquired as cellulose or

lignin by their host termites and metabolises the carbohydrate into acetic acid which is
utilized back by the termites.
(iii) Enteric bacteria - Human gut

● The relationship between some gut flora and humans is rather a mutualistic
relationship.
● Some human gut microorganisms benefit the host by fermenting dietary fiber into
short-chain fatty acids (SCFAs), such as acetic acid and butyric acid, which are then
absorbed by the host.
● Intestinal bacteria also play a role in synthesizing vitamin B and vitamin K as well as
metabolizing bile acids, sterols, and xenobiotics (a chemical substance found within an
organism that is not naturally produced or expected to be present within the organism).
(iv) Mycorrhiza :
● This is a symbiotic association between a green plant and a fungus.
● The plant makes organic molecules such as sugars by photosynthesis and supplies them
to the fungus, and the fungus supplies to the plant water and mineral nutrients, such as
phosphorus, taken from the soil.
(v) Paramecium-Chlorella:
● Paramecium (protozoa) can host Chlorella (algae) within its cytoplasm.
● The algae Chlorella provides the protozoan partner with carbon and oxygen, in turn
protozoa provide protection, motility, carbondioxide and other growth factors.
● The presence of Chlorella within Paramecium helps to survive protozoa in anaerobic
condition as long as there is sufficient light.
(2) SYNTROPHISM :
● It is an association in which the growth of one organism either depends on or improved
by the substrate provided by another organism.
● In syntrophism both organism in association get benefits.
Examples of syntrophism:
(i) Lactobacillus arobinosus and Enterococcus faecalis:

● In the minimal media, Lactobacillus arobinosus and Enterococcus faecalis are able to
grow together but not alone.
● The synergistic relationship between E. faecalis and L. arobinosus occurs in which E.

faecalis require folic acid which is produced by L. arobinosus and in turn lactobacillus
require phenylalanine which is produced by Enterococcus faecalis.
(3) PROTO-COOPERATION
● It is a relationship in which organisms in association mutually benefits each other.
● This interaction is similar to mutualism but the relationship between the organisms in
protocooperation is not obligatory as in mutualism.
Examples of Protocooperation:
(i) Association between Ants and Aphids

● A further example of protocooperation is the connection between ants and aphids.
● The ant searches for food on trees and shrubs that are hosts to honeydew-secreting
species such as aphids. The ant gathers the sugary substance and takes it to its nest as
food for its offspring.
● It has been known for the ant to stimulate the aphid to secrete honeydew straight into
its mouth. Some ant species even look after the honeydew producers from natural
predators.
● In areas where the ant inhabits the same ecosystem as the aphid, the plants they inhabit
normally suffer from a higher presence of aphids which is detrimental to the plant but
not to the two species protocooperating.
(4) COMMENSALISM
● It is a relationship in which one organism (commensal) in the association is benefits

while the other organism (host) of the association neither benefits nor harmed
● It is an unidirectional association and if the commensal is separated from the host, it can
survive.
Examples of commensalism:
(i) Non-pathogenic coli in gut of human:

● Escherichia coli is a facultative anaerobe that uses oxygen and lower the oxygen
concentration in gut which creates suitable environment for obligate anaerobes such as
Bacteroides.
● E. coli is a host which remains unaffected by Bacteroides.
(ii) Association of Nitrosomonas (host) and Nitrobacter (commensal) in Nitrification:

● Nitrosomonas oxidize Ammonia into Nitrite and finally Nitrobacter uses nitrite to obtain
energy and oxidize it into Nitrate
NEGATIVE INTERACTIONS
(1) AMENSALISM :
● When one microbial population produces substances that is inhibitory to other

microbial population then this inter population relationship is known as Ammensalism
or Antagonism.
● It is a negative relationship.
● The first population which produces inhibitory substances are unaffected or may gain a
competition and survive in the habitat while other population get inhibited. This
chemical inhibition is known as antibiotics.
Examples of amensalism :
(i) Lactic acid produced by lactic acid bacteria in vaginal tract:

● Lactic acid produced by many normal floras in vaginal tract is inhibitory to many
pathogenic organisms such as Candida albicans.
(ii) Skin normal flora:

● Fatty acid produced by skin flora inhibits many pathogenic bacteria on the skin
(iii) Thiobacillus thiooxidant:

● Thiobacillus thioxidant produces sulfuric acid by oxidation of sulfur which is responsible
in lowering of pH in the culture media which inhibits the growth of most other bacteria.
(2) COMPETITION :
● The competition represents a negative relationship between two microbial population in

which both populations are adversely affected with respect to their survival and growth.
● Competition occurs when both population uses same resources such as space or
nutrition, so, the microbial population achieves lower maximum density or growth rate.
● Microbial population competes for any growth limiting resources such as carbon source,
nitrogen source, phosphorus, vitamins, growth factors etc.
● Competition inhibits both population from occupying exactly same ecological niche
because one will win the competition and the other one is eliminated.
Examples of competition:
(i) Competition between Paramecium cadatum and Paramecium aurelia:

● Both species of Paramecium feeds on same bacteria population when these protozoa
are placed together.
● P. aurelia grow at better rate than P. caudatum due to competition.

(3) PARASITISM
● It is a relationship in which one population (parasite) get benefited and derive its
nutrition from other population (host) in the association which is harmed.
● The host-parasite relationship is characterized by a relatively long period of contact

which may be physical or metabolic.
● Some parasite lives outside host cell, known as ectoparasite while other parasite lives
inside host cell, known as endoparasite.
Examples of parasitism:
(i) Viruses:
● Viruses are obligate intracellular parasite that exhibits great host specificity.
● There are many viruses that are parasitic to bacteria (bacteriophage), fungi, algae,
protozoa etc.
(ii) Bacteria :
● Shigella dysenteriae invades and exist parasitically within human host causing
dysentery.
(iii) Protozoa :
● Plasmodium malariae introduced into a human host via a female anopheles mosquito
bite exist parasitically causing malaria.
(4) PREDATION :
● It is a wide spread phenomenon when one organism (predator) engulfs, and attacks
another organism (prey).
● The prey can be larger or smaller than the predator and this normally results in death of
prey.
● Normally predator-prey interaction is of short duration.

Examples of Predation:
(i) Protozoan-bacteria in soil:

● Many protozoans can feed on various bacterial population which helps to maintain
count of soil bacteria at optimum level.
(ii) Bdellovibrio, Vamparococcus, Daptobacter etc are examples of predator bacteria that can
feed on wide range of bacterial population.
7.4 ECONOMIC IMPORTANCE OF MICROORGANISMS
● They are useful in the baking industry for the production of bread etc E.g.
Saccharomyces cerevisea
● They are useful in environmental bioremediation E.g. Pseudomonas putida
● They facilitate spoilage of food substances. E.g. Mucor
● They are useful in genetic engineering for the production of vaccine E.g. Polio vaccine
● In yoghurt production, microorganism serves as probiotics. E.g. Lactobacillus sp
● They facilitate medical advancement by the production of penicillin. E.g. Penicillium

notatum
● They are important disease causing entities to both man and livestock. E.g. Syphilis
(Treponema pallidum), Anthrax (Bacillus anthracis).
● Salmonella enterica exists in a mutualistic association in the gastrointestinal tract of

human assisting in the digestion of dietary fibre and synthesis of vitamin B and K.
● Lactobacillus the microflora of the vagina maintains the acidity of the vagina thereby
preventing the survival of pathogen such as Candida albicans.
● Fungi are considered very important in the production of certain enzymes useful by man
such as lysozyme.
● The fungi partner in mycorrhiza association supplies the plant with plant water and
mineral nutrients, such as phosphorus, taken from the soil thus enhancing the quality
plants.
7.5 METHODS OF MICROBIAL CONTROL
MOIST HEAT RADIATION DRY HEAT
Pasteurization Ionizing of Xray Incineration
Autoclaving Non-ionizing of UV rays Dry heat oven and Filtration.

MULTIPLE CHOICE
1. Ecology can be defined as

(a) The scientific study of living organism and their external environment of matter and
energy.
(b) A self sustaining unit made up of biotic and abiotic components
(c) The study of the relationship between living organisms and their environment.
(d) The study of the relationship of microorganisms with each other and their
environment.
2. Which best defines an ecosystem

energy.
environment.
3. Which best describes microbial ecology

energy.
environment.
4. The association between ants and aphids is best describes as

(a) Mutualism.
(b) Proto coperation
(c) Commensalism
(d) Parasitism
5. The association between varicella virus found in the blood of humans is

(a) Mutualism.
(c) Commensalism
(d) Parasitism
6. The association between Nitrosomonas and Nitrobacter is best described as

(a) Mutualism.
(c) Commensalism
(d) Parasitism
7. The association between flagellated protozoan and termite is

(a) Mutualism.
(c) Commensalism
(d) Parasitism
8. Organisms that utilises sunlight to produce energy are called. (a)Autotrophs

(b)Lithotrophs
(c)Photoautotrophs
(d) heterotrophs
9. The complete breakdown of organic molecules into inorganic molecules Is termed.

(a) mineralization (b) Eutrophication. (c)Erosion (d) Hydrolysis
10. Organisms that digest the dead remains of primary producers and consumers are called.
(a) Heterotrophs. (b)Decomposers. (d)Consumers. (d) Autotrophs
11. The microflora of the vagina which maintains its acidity thereby preventing the survival
of pathogen is (a) Lactobacillus sp. (b)Candida albicans (d) Shigella sp. (d)
Salmonella enterica
12. Malaria is caused by

(a) Introduction of Plasmodium malariae into a human host by a mosquito bite
(b) Consumption of contaminated food and water
(c)Introduction of Plasmodium malariae into a human host by a tse-tse fly bite
(d) Introduction of Plasmodium malariae into a human host by a female anopheles
mosquito bite
13. The algal partner in lichen is called. (a)Phycobiont (b) Ectosymbiont.
(d)Mycobiont. (d) Endosymbiont
14. The production of penicillin an antibiotic is peculiar to (a)Penicillium notatium. (b)

Treponema pallidum (d) Shigella sp. (d) Penicillium notatum
15. The causal organism of dysentery is (a)Treponema pallidum. (b)Candida albicans (d)
Shigella dysenteriae. (d) Salmonella enterica
16. Which of the following is a negative relationship. (a) Amensalism (b)Commensalism

(d)Mutualism. (d) Proto coperation
17. Which of the following is a positive relationship. (a) Commensalism (b)Predation (d)
Parasitism. (d) Ammensalism
18. In baking and brewery industry, an important microorganism in production is (a)

Saccharomyces cerevisea (b) Staphylococcus aureus (d) Streptococcus pyogene (d)
Salmonella enterica
19. The causal organism of sore throat in human is. (a) Saccharomyces cerevisea (b)
Staphylococcus aureus (d) Streptococcus pyogene (d) Salmonella enterica
20. A relationship in which one organism benefits while the other neither benefits nor
harmed is
(a) Commensalism (b)Predation (d) Parasitism. (d) Ammensalism
21. From the question above, the benefitted organism is called the (a) Commensal
(b)Predator (d) Parasite (d) Ammensal
22. Organisms that utilises inorganic substances to derive energy are called. (a)Autotrophs
(b)Chemolithotrophs
(c)Photoautotrophs
(d) heterotrophs
23. The fungal partner in lichen is called. (a)Phycobiont (b) Ectosymbiont.

(c)Mycobiont. (d) Endosymbiont
24. A relationship in which one organism benefits from and harms the other organism is.
(a) Commensalism (b)Predation (c) Parasitism. (d) Ammensalism
(b)Predator (c) Parasite (d) Ammensal
26. A relationship whereby one microbial population produces substances that is inhibitory
to other microbial population is (a) Commensalism (b)Predation (c) Parasitism. (d)
Ammensalism
(b)Predator (c) Parasite (d) Ammensal
28. A relationship that exist between a smaller organism located on the surface of a larger
organism is (a) Ectosymbiosis. (b)Synergism (c) Endosymbiosis (d) Symbiosis
29. The smaller organism is called the

(a) Ectosymbiont. (b)Synergist (c) Endosymbiont (d) Symbiont
30. A relationship that exist between a smaller organism located inside a larger organism is
(a) Ectosymbiosis. (b)Synergism (c) Endosymbiosis (d) Symbiosis
31. The smaller organism is called the

(a) Ectosymbiont. (b)Synergist (c) Endosymbiont (d) Symbiont
32. The association between Lactobacillus arobinosus and Enterococcus faecalis is.
(a) Syntrophism (b)Mutualism. (C) Commensalism (d)Parasitism
33. A chemical substance found within an organism that is not naturally produced or
expected to be present within the organism is (a) Xenobiotics (b)Probiotics. (c)
Toxins (d)None of the above
34. Lichen is an association made up of (a) Fungi and Algae

(b) Fungi and photosynthetic algae
(c) Fungi and protozoan
(d) Protozoan and photosynthetic algae.
35. A Fungi that specializes in food spoilage is (a) Mucor. (b)Escherichia coli
(c)Pseudomonas sp.
(d) Clostridium sp
36. The region of the earth inhabited by life is (a)Biosphere (b) Lithosphere.
(c)Hydrosphere. (d) Atmosphere
37. The region of the earth occupied by rocks and mountain is (a)Biosphere
(b) Lithosphere. (c)Hydrosphere. (d) Atmosphere
38. The region of the ecosystem occupied by water is (a)Biosphere (b)
Lithosphere. (c)Hydrosphere. (d) Atmosphere
39. The region of the ecosystem occupied by gas is (a)Biosphere (b)
Lithosphere. (c)Hydrosphere. (d) Atmosphere
40. A significant microbe in bioremediation is. (a) Pseudomonas putida.
(b) Staphylococcus aureus (c) Streptococcus pyogene (d) Salmonella
enterica
THEORY
1. Define the following.
(i) Hydrosphere
(ii) Atmosphere
(iii) Lithosphere
(iv) Ecosphere
(v) Biosphere
2. Differentiate between the following pair

(i) Ecology and microbial Ecology
(ii) Ecosystem and Environment
(iii) Commensalism and Mutualism
(iv) Parasitism and Ammensalism
(v) Competition and Predation
(vi) Proto cooperation and Syntrophism
(vii) Ectobiont and Endobiont
(viii) Phycobiont and Mycobiont
3. Enumerate 10 economic importances of microorganisms
4. Differentiate between Producers Consumers and Decomposers
5. Enumerate 10 medically important microorganism
6. Highlight various methods of microbial control.

1. C
2. B
3. D
4. B
5. D
6. C
7. A
8. C
9. A
10. B
11. A
12. D
13. A
14. D
15. D
16. A
17. A
18. A
19. D
20. A
21. A
22. B
23. C
24. C
25. C
26. D
27. D
28. A
29. A
30. C
31. C
32. A
33. A
34. B
35. A
36. A
37. B
38. C
39. D
40. A
THEORY

2. Make reference to Chapter 7.1,2,3
5. Penicillium notatum produces antibiotic penicillin.
Treponema pallidum causes syphilis
Streptococcus pyogene causes sore throat.
Varicella zoster virus causes chicken pox
Lactobacillus sp maintains acidity of the vagina thereby preventing any pathogenic

invasion.
Salmonella enterica aids in fiber and cellulose digestion in man thereby preventing
constipation.
Staphylococcus aureus exists and secretes useful substances on the skin which prevent
skin burn from UV light and survival of pathogens.
Polio virus causes poliomyelitis
Plasmodium malariae causes malaria
Candida albicans causes vaginal candidiasis.
CHAPTER 8
8.1 NATURE OF GENES.

In 1953, Watson and crick proposed the helical structure of DNA
saying that DNA controls the activity of a living organism.
DNA (Deoxyribose-Nucleic-Acid) is made up of double strands of Deoxyribose sugar, phosphate
group, Nitrogenous bases.
While RNA (Ribose-Nucleic-Acid) is made up of single strands of Ribose sugar, phosphate

group, and Nitrogenous bases.
The nitrogenous base could be purines (Double ring structure which are Adenine A and
Guanine G), or pyrimidine (Single ring structures which are Cytosine C and Thiamine T or Uracil
U in RNA).
Adenine binds to Thiamine in DNA but binds to Uracil in RNA

A-----T (DNA). A-----U (RNA)
While Cytosine binds to Guanine in both DNA and RNA

C-----G
8.2. CODON
Codon also known as genetic code can be defined as the triplet code of nucleotide. It's also
defined as a collection of base sequences that corresponds to each Amino acid.
Example: Codon UUU codes for Phenylalanine

UUA codes for LEUCINE
AUG codes for METHIONINE
GUC codes for VALINE
GCU codes for ALANINE
Triplet of nucleotide that is complimentary to the codon is called anticodon

Example CAU---RNA---(codon)
GTA---DNA---(anticodon)
When there's an alteration in the Nitrogenous base sequence in a codon, mutation has
occurred.
There are 64 genetic codes of which 3 are non-sense codon (UAG, UAA, UGA) because they
code for nothing while 61 are sense codon.
CHARACTERISTICS OF GENETIC CODE

● It's a triplet codon
● The code is non over lapping
● It is universal meaning it's the same in all organisms
● It's comaless meaning it's continuous
● It's degenerative meaning an Amino acid can be coded for by more than one code
except methionine and tryptophan.
● Some codon are known as start codon e.g. AUG
● Some codon act as stop codon e.g. UAA, UGA, UAG
8.3 MUTATION
Mutation is a sudden change in the arrangement, amount, structure of the DNA of an organism
which results in the formation of a new character that can be inherited.
Mutation can also occur when the nucleotide of the DNA undergoes changes in helical structure
resulting in change of the DNA and production of organism with different property.
TYPES OF MUTATION
● Point mutation is a change in single base in a codon e.g. UUC becomes UUA, the
mutation occurs at the last nitrogenous base. This is point mutation.
● Spontaneous Mutation is a type of mutation that occurs naturally in the environment;
but slowly; its estimated at 10-4 - 10-12 in each cell division.
Probability that mutation will occur in a given gene in each cell division is called rate of
mutation estimated at 10-4 to 10-12.
● Induced Mutation is a type of mutation formed as a result of exposure of a cell to a
mutagen; either chemical or radiation.
Mutagen is any substance or agent that causes mutation
Carcinogen is any substance or agent that causes cancer e.g ethylenebromide.
● Base substitution which includes
-Point Mutation: change in single base.
- Miscense mutation: mutation as a result of wrong substitution of amino acid e.g replacement
of purine by purine.
When a purine is replaced by a purine is called transition.
When a purine is replaced by a pyrimidine is called transversion.
-Silent mutation: a situation where mutation takes place but it's not conspicuous
phenotypically because the same amino acid is still being produced e.g {CUU,CUC}leucine to
{CUC,CUC} still leucine.
It is a type of mutation that occurs when there are more than one codon coding for one amino
acid, and one base is replaced by another, both still coding for the same amino acid , though
mutation has taken place genotypically, but the phenotypic appearance is still the same.
-Nonsense mutation: A mutation that changes a codon that normally codes for amino acid to a
stop codon e.g UAC to UAG
● Frameshift Mutation occurs as a result of addition or removal of a nucleotide in DNA.
● Transposable Element Also reffered to as insertion element or insertion sequence or
transposome. They are special segment of DNA that can move from one region of
chromosome to another. Thus they are called Jumping gene.
8.4. CHEMICAL MUTAGEN

HNO2 (Nitrous fluid)- Converts the NH3 content of amino acid into keto group. The implication
is that it will convert cytosine to uracil causing Cytosine to bond with adenine instead of
guanine.
Alkylating agent - E.g Nitrosoguanidine which adds short chains of methyl group to either
purine or pyrimidine.
Base Analog- They are intercalating agent compound that have structure similar to parent
structure. The implication of this is that it alters hydrogen bonding of that compound.
Ethylium bromide is both a mutagen and carcinogen.
8.5. PHYSICAL AGENT (MUTAGEN)

Radiation(UV and X-ray)
● UV: non- ionising radiation
● X-ray: ionising radiation
Formation of thymine dimer is the implication of exposure of cell to UV light.
NB: Organisms that does not show the effect of mutation are called the wild type.
While organisms or offsprings showing the effect of mutation are called mutants.
The progeny of a mutant is also a mutant.
8.6 TRANSFER OF MUTATION
Mutation passed from parent to progeny is called vertical transfer of genes.
While transfer of gene from one cell to another mediated by plasmid is called horizontal gene
transfer.
Recall that plasmids are extra chromosomal DNA molecule capable of autonomous replication
and independent of the host DNA.
HORIZONTAL GENE TRANSFER

There are 3 methods
•Transformation: This occurs when naked DNA is transferred from cell to environment and
then picked up by another cell.
•Conjugation: This occurs when there is transfer of genetic material from donor to recipient
when in contact e.g two bacteria lie side by side and joined by sex pillus and DNA is transferred
through sex pillus from donor to recipient
•Transduction: This occurs when there is transfer of DNA through bacteriophage from donor
cell to recipient cell.
WHY BACTERIA ARE USED IN GENETIC STUDY.

● They are abiquitous
● Can be grown in an inexpensive culture media
● Have fast replication rate
● Can easily be subjected to genetic manipulation
8.7 NUCLEOSIDE & NUCLEOTIDE

Nucleoside is the association between nitrogenous base and pentose sugar. Depending on the
type of Nitrogenous bases and pentose sugar, there are different types of nucleoside each with
it's own characteristic and structure.
Nucleotide is the association between nitrogenous bases, pentose sugar and phosphate group.
The phosphate group which is the distinguishing factor between nucleotide and nucleoside
connects one pentose sugar to another, thereby forming a strand.
8.8 RNA
The Ribose nucleic acid has 4 types, each enclosed by it own type of gene. The genomic RNA
contains all the information for the structure and function of the organism.
MESSENGER RNA (mRNA)

● It encodes amino acid sequence of polypeptides
TRANSFER RNA (tRNA)

● Brings amino acid to the ribosome during translation
RIBOSOMAL RNA (rRNA)

● Together with ribosomal protien, they make up the ribisome, the organelle that
translates the mRNA.
● Therefore rRNA + Ribosomal protein = Ribosome
SMALL NUCLEAR RNA (Sn RNA)

● Together with proteins, they form complexes that are used in RNA processing in
eukaryotic cell but not found in prokaryotes.
● SnRNA + proteins = Complexes used in RNA processing in eukaryotes.
TRANSCRIPTION: A process by which the DNA is being copied to form RNA using enzyme RNA
polymerase.
Transcription is the first step in gene expression in which a particular segment of DNA is copied
into RNA (especially mRNA) by an enzyme RNA polymerase.
Transcription is divided into 4 stages.
● Initiation: Transcription begins with the binding of RNA polymerase with one or more
transcription factor to a specific DNA sequence called promoter to form RNA
polymerase promoter-complex.
● Promoter Escape: After the first step is synthesised, the RNA polymerase enzyme must
escape the promoter.
● Elongation: One strand of DNA ie the template strand (non coding strand) is used as a
template for RNA synthesis.
● Termination: In bacteria, it could be by role dependent or role independent.
TRANSLATION: A process by which the information in RNA is copied to form protein.

It involves decoding a messenger RNA and using it's information to build a polyeptide or a chain
of amino acid.
Translation begins with codon (AUG) Methionine. There are about 6 codon for amino acid.
3 stop codon marks the end of polypeptide.
Steps in translation
Initiation
Elongation
Termination
8.9 REPLICATION OF DNA

Deoxyribonucleic acid is a molecule composed of two polynucleotide chains that coil around
each other to form a double helix carrying genetic instructions for the development,
functioning, growth and reproduction of all known organisms and many viruses.
Replication of DNA is a process by which the DNA double helix is unwound and parent DNA is
copied to produce daughter strands with the aid of various enzymes.
The goal of DNA replication is to make a second DNA molecule, using the parent strands as a
template to create two new daughter strands.
In this process, both parent strands are conserved, or saved, in each of the new molecules. This
is called semi-conservative replication.
In semi-conservative replication, the parent strands split apart, but each remains whole, while
new daughter strands are added onto them. The end result is two DNA molecules, each
containing one parent strand and one daughter strand.
STEPS AND ENZYMES IN DNA REPLICATION

First step
● DNA helicase unwinds the double helix by breaking the hydrogen bonds between the
parent strands of DNA. This splitting and unwinding process opens up the DNA molecule
into a Y-shape, which is called the replication fork
● DNA helicase is the enzyme that unwinds the DNA double helix.
Second step
● Before new daughter strands can be added on, the parent strands are first made ready
by an RNA primer. The primer is built by the enzyme RNA primase.
● RNA primase is the enzyme that builds an RNA primer on the parent strand to initiate
DNA replication.
Third step
● Once the RNA primer is built, DNA polymerase slides into the replication fork and
positions itself behind the RNA primer. It begins to add DNA nucleotides onto each
parent strand.
● DNA polymerase always works by starting at the 3' end of DNA and moving toward the
5' end. This means that on the leading strand, it works continuously as it follows DNA
helicase, which is constantly opening the fork more and more.
● But on the lagging strand, DNA polymerase works discontinuously, making Okazaki
fragments in the opposite direction.
● DNA polymerase is the enzyme that matches and lays down nucleotides to build the
daughter DNA strand along each parent DNA strand.
Fourth step
● Okazaki fragments that are separate from each other, are joined together by the
enzyme DNA ligase.
● Okazaki fragments are short sequences of DNA nucleotides which are synthesized
discontinuously and later linked together by the enzyme DNA ligase.
● DNA ligase binds the fragments end to end, forming a continuous daughter strand on
top of the lagging parent strand.
● DNA ligase is an enzyme that joins the adjacent Okazaki fragments on the lagging
strand of DNA.
8.10 BIOCHEMICAL TEST

MOTILITY TEST
Procedure
● Innoculate a medium with 18-24hrs culture by stab innoculation using innoculation
needle.
● Incubate the medium at room temperature
● Examine tubes for growth and signs of motility.
● Motility is positive if there is presence of growth away from the line of innoculation
while non motile organism only grow along the line of ininnoculation.
CATALASE TEST
Procedure
● Transfer a well isolated cell to a clean glass slide and add a drop of hydrogen peroxide
(H2O2) without mixing.
● Observe for immediate formation of gas bubbles.
Result
● Formation of gas bubbles indicate positive result.
OXIDASE TEST
Procedure
● Add few drops of oxidase test reagent to a strip of filter paper (whatman filter paper).
● Streak a loop full of bacteria on to the reagent saturated filter paper with a wooden
applicator stick.
Result
● Positive reaction turns the bacteria from violet to purple within 30secs, while negative
remains colourless.
COAGULASE TEST
Procedure
● A loop full of isolated bateria from a 18-24hrs culture is emulsified on a slide with
normal saline.
● A drop of undiluted plasma is added to the suspension and stired for 5secs.
Result
● Clumping of colonies indicate a coagulase positive result

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Microbiology

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(Author : Umossoh A. A.

1.1 INTRODUCTION TO MICROBIOLOGY

● Virology - Study of viruses, its structures, functions and classification.

● Mycology - Study of fungi, its structures, functions and classification.

● Bacteriology - Study of bacteria, its structures, functions and

● Protozoology - Study of protozoa, its structures, functions and

1.2 TERMS IN MICROBIOLOGY

Bactericidal – A substance of chemical complexity that kills bacteria out-rightly.

Germicide – Any substance that kills germs.

Sterile – A state of total absence of microorganism on a surface.

Immunity – A state of non-susceptibility of an organism to diseases.

Contamination – The presence of constituent impurities and unwanted elements on a

Decontamination – A process of treatment that renders a body surface safe to handle by

Culture- A collection or population of microbial growth in a nutrient media.

Recombinant: This refers to something formed by combining existing elements in a new

1.3 CONTRIBUTION OF SCIENTISTS TO MICROBIOLOGY

Louis Pasteur a French chemist and microbiologist is considered “Father of Modern

Alexander Fleming a Scottish bacteriologist accidentally discovered the antibiotic in 1928,

1.4 HISTORY OF THE DISCOVERY OF MICRO-

▪ Conflict Arising from Spontaneous Generation

Lazzaro Spallanzani (1751): Disagreed with Needham’s conclusion through carefully

ANALOGY OF REDI’S EXPERIMRNT (SOURCE: LUMENLEARNING.COM)

1.5 GERM THEORY OF DISEASE

SELF ASSESSMENT QUESTIONS

9. According to Koch’s postulate, arrange the following in the right order

11. Which of the following is incorrect

13. A Pandemic can be accurately defined as (a) The

14. The Germ theory of diseases is accredited to (a) Louis Pasteur

19. Which of the following is incorrect (a)Epidemic – The outbreak of disease in a

28. Agar can be defined as (a) A solidifying agent usually introduced to

SELF ASSESSMENT QUESTIONS

5. Differentiate between the Theory of equivocal generation and Theory of univocal

ANSWERS (Multiple Choice)

4. (a) Malaria (b) A bite by an infected female anopheles mosquito. (c)

2.1 TYPES AND TAXANOMIC GROUPINGS OF

2.2 DIFFERENCES BETWEEN PROKARYOTES & EUKARYOTES

It lacks a membrane bound nucleus, thus It has a well-defined membrane bound

It possess 70s ribosome. It possess 80s ribosome.

In photosynthetic cells, chlorophll is located In photosynthetic cells, chlorophyll is located

Their chromosomes are restricted to the Their chromosomes is restricted to the

SIMILARITIES BETWEEN PROKARYOTES & EUKARYOTES.

● They both possesses genetic materials.

● They both have a cytoplasm.

● They both have a plasma or cell membrane.

● They both possesses ribosomes.

● They both possesses enzymes.

STUCTURE OF AN ENVELOPED VIRUS (SOURCE: BIOLOGY DICTIONARY.NET)

A Prion is a proteinaceous infectious particle that is responsible for at least 7 neuro-

2.4 VIRAL CLASSIFICATION

2.5 SHAPES OF VIRUS

2.7 REPLICATION CYCLE OF VIRUSES IN HOST CELL.

REPLICATION CYCLE OF A VIRUS IN A HOST CELL (SOURCE: SPARKNOTE LLC)

2.8 FUNCTIONS OF THE STRUCTURES OF A VIRUS

ANALYSIS OF VIRAL GENOME. (SOURCE: SCIENCE DIRECT)

● They are capable of mutation.

● The shape of a virus is determined by the shape of the capsid.

● Capsids are composed of many identical protein subunits called capsomeres.

2.10 VIRAL DISEASES, CAUSATIVE AGENTS AND MODE OF INFECTION

Small pox Variola Virus ● Person to person

Yellow fever Flavi Virus ● Infected mosquitoes

Rabbies Rhabdo Virus ● Virus is usually

Ebola Virus Disease (EVD) Ebola Virus ● Ingestion of raw or

Chicken pox Varicella zoster Virus ● Mostly airborne,

Hepatitis B Hepatitis B Virus (HBV) ● Perinatal transmission