MICB 201

GENERAL MICROBIOLOGY I

BY
Dr. SHU’AIBU ISA

COURSE OUTLINE
Spontaneous generation and fermentation; Scope of microbiology, general
characteristics of microorganisms, growth and reproduction of microbes’ brief survey of
microbes as friends and foes; Nutritional and biochemical activities of microorganism,
Antigens and antibodies; Identification and economic importance of selected microbial
groups

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 SPONTANEOUS GENERATION

The Theory of Spontaneous Generation

Humans have for a number of millennia been asking: 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.

The Greek philosopher Aristotle (384–322 BC) was one of the earliest recorded
scholars to articulate the theory of spontaneous generation, the notion that life can
arise from nonliving matter. Aristotle proposed that life arose from nonliving material if
the material contained pneuma (“vital heat”). As evidence, he noted several instances of
the appearance of animals from environments previously devoid of such animals, such
as the seemingly sudden appearance of fish in a new puddle of water.

This theory persisted into the seventeenth century, when scientists undertook additional
experimentation to support or disprove it. By this time, the proponents of the theory
cited how frogs simply seem to appear along the muddy banks of the Nile River in Egypt
during the annual flooding. Others observed that mice simply appeared among grain
stored in barns with thatched roofs. When the roof leaked and the grain molded, mice
appeared. Jan Baptista van Helmont, a seventeenth century Flemish scientist, proposed
that mice could arise from rags and wheat kernels left in an open container for 3 weeks.
In reality, such habitats provided ideal food sources and shelter for mouse populations
to flourish.

However, one of van Helmont’s contemporaries, Italian physician Francesco Redi

(1626–1697), performed an experiment in 1668 that was one of the first to refute the
idea that maggots (the larvae of flies) spontaneously generate on meat left out in the

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open air. He predicted that preventing flies from having direct contact with the meat
would also prevent the appearance of maggots. Redi left meat in each of six containers
(Figure 1). Two were open to the air, two were covered with gauze, and two were tightly
sealed. His hypothesis was supported when maggots developed in the uncovered jars,
but no maggots appeared in either the gauze-covered or the tightly sealed jars. He
concluded that maggots could only form when flies were allowed to lay eggs in the
meat, and that the maggots were the offspring of flies, not the product of spontaneous
generation.

Figure 1: Francesco Redi’s experimental setup consisted of an open container, a container sealed with a
cork top, and a container covered in mesh that let in air but not flies. Maggots only appeared on the
meat in the open container. However, maggots were also found on the gauze of the gauze-covered
container.

In 1745, John Needham (1713–1781) published a report of his own experiments, in

which he briefly boiled broth infused with plant or animal matter, hoping to kill all
preexisting microbes. He then sealed the flasks. After a few days, Needham observed
that the broth had become cloudy and a single drop contained numerous microscopic
creatures. He argued that the new microbes must have arisen spontaneously. In reality,
however, he likely did not boil the broth enough to kill all preexisting microbes.

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Lazzaro Spallanzani (1729–1799) did not agree with Needham’s conclusions, however,
and performed hundreds of carefully executed experiments using heated broth. As in
Needham’s experiment, broth in sealed jars and unsealed jars was infused with plant
and animal matter. Spallanzani’s results contradicted the findings of Needham: Heated
but sealed flasks remained clear, without any signs of spontaneous growth, unless the
flasks were subsequently opened to the air. This suggested that microbes were
introduced into these flasks from the air. In response to Spallanzani’s findings,
Needham argued that life originates from a “life force” that was destroyed during
Spallanzani’s extended boiling. Any subsequent sealing of the flasks then prevented
new life force from entering and causing spontaneous generation (Figure 2).

Figure 2: (a) Francesco Redi, who demonstrated that maggots were the offspring of flies, not products
of spontaneous generation. (b) John Needham, who argued that microbes arose spontaneously in broth
from a “life force.” (c) Lazzaro Spallanzani, whose experiments with broth aimed to disprove those of
Needham.
The debate over spontaneous generation continued well into the nineteenth century,
with scientists serving as proponents of both sides. To settle the debate, the Paris
Academy of Sciences offered a prize for resolution of the problem. Louis Pasteur, a
prominent French chemist who had been studying microbial fermentation and the
causes of wine spoilage, accepted the challenge. In 1858, Pasteur filtered air through a
gun-cotton filter and, upon microscopic examination of the cotton, found it full of
microorganisms, suggesting that the exposure of a broth to air was not introducing a
“life force” to the broth but rather airborne microorganisms.

Later, Pasteur made a series of flasks with long, twisted necks (“swan-neck” flasks), in

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which he boiled broth to sterilize it (Figure 3). His design allowed air inside the flasks to
be exchanged with air from the outside, but prevented the introduction of any airborne
microorganisms, which would get caught in the twists and bends of the flasks’ necks. If
a life force besides the airborne microorganisms were responsible for microbial growth
within the sterilized flasks, it would have access to the broth, whereas the
microorganisms would not. He correctly predicted that sterilized broth in his swan-neck
flasks would remain sterile as long as the swan necks remained intact. However, should
the necks be broken, microorganisms would be introduced, contaminating the flasks
and allowing microbial growth within the broth.

Pasteur’s set of experiments irrefutably disproved the theory of spontaneous generation

and earned him the prestigious Alhumbert Prize from the Paris Academy of Sciences in
1862. In a subsequent lecture in 1864, Pasteur articulated “Omne vivum ex vivo” (“Life
only comes from life”). In this lecture, Pasteur recounted his famous swan-neck flask
experiment, stating that “life is a germ and a germ is life. Never will the doctrine of
spontaneous generation recover from the mortal blow of this simple experiment.”[4] To
Pasteur’s credit, it never has.

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Figure 3: (a) French scientist Louis Pasteur, who definitively refuted the long-disputed theory of
spontaneous generation. (b) The unique swan-neck feature of the flasks used in Pasteur’s experiment
allowed air to enter the flask but prevented the entry of bacterial and fungal spores. (c) Pasteur’s
experiment consisted of two parts. In the first part, the broth in the flask was boiled to sterilize it. When
this broth was cooled, it remained free of contamination. In the second part of the experiment, the flask
was boiled and then the neck was broken off. The broth in this flask became contaminated.

SCOPE OF MICROBIOLOGY

Microbiology is simply defined as the study of microorganisms. However,

microorganisms are living organisms that cannot be seen by an unaided eye. They can
only be seen with the seen with equipment called microscope. These microorganisms
include Viruses, bacteria, fungi, protozoa and some algae. Microbiology is divided into
two branches namely Basic or Pure microbiology and Applied microbiology. The basic
microbiology is defined as the basic aspects of microbiology and is further subdivided
into Organism-centered (Taxonomic arrangement) and Process centered or
(Integrative arrangement). However, the applied microbiology involves the application

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of microorganisms for human use and consists of the following of the following
branches; Medical Microbiology, Public Health Microbiology, Pharmaceutical
Microbiology, Veterinary Microbiology, Industrial Microbiology, Food and Dairy
Microbiology, Agricultural Microbiology, Environmental Microbiology, Aero Microbiology,
Microbial Biotechnology, Water and Aquatic Microbiology, Vaccinology and
Chemotherapy.

Basic or Pure Microbiology

This involves the basic aspects of microbiology and involves the following branches

(1) Organism-centered or Taxonomic arrangement: This is a branch of basic

microbiology that focuses on the study of microorganisms based on their
taxonomy. It is further divided into:

(a) Virology: The study of viruses

(b) Bacteriology: The study of bacteria

(c) Mycology: The study of fungi (yeasts and molds)

(d) Phycology: The study of algae

(e) Protozoology: The study of protozoans

(f) Helminthology: The study of helminthes such as nematodes, round and filarial
worms

(g) Immunolgy: The study of microorganisms in relation to immune system

(2) Process centered or (Integrative arrangement): This is a branch of basic

microbiology that focuses on the study of microorganisms in relation to their
characteristics. It is also divided into:

(a) Microbial Cytology: This is concerned with the structure and function of
microbial cells.

(b) Microbial Physiology: The study of how microbial structures, growth and

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 metabolism function in living organisms.

(c) Microbial Genetics: The study of the mechanisms of heritable information in

microorganisms.

(d) The study that explores the diversity, distribution and abundance of
microorganisms, their specific interactions, and the effect that they have an
ecosystems.

(e) Microbial Taxonomy: This branch studies a means by which microorganisms

can be grouped together

(f) Molecular Microorganisms: This deals with the molecular mechanisms and
physiological processes of microorganisms and their utilization in production
of biotechnology products and medicines such as vaccines, antibodies etc. It
also involves advancement in pathogenicity of microorganisms.

(g) Cellular Microbiology: This is the study of the functions and properties of
microbial cells.

Applied Microbiology

This involves the application aspects of microbiology and involves the following
branches

(1) Medical Microbiology: Also known as clinical microbiology is a sub-discipline of

microbiology dealing with the study of microorganisms (parasites, fungi, bacteria,
viruses, and prions) capable of infecting and causing diseases in humans.

(2) Public Health Microbiology: Public Health microbiology is a microbiology that

spans the fields of human, animal, food, water and environmental microbiology,
with a focus on human health and disease.

(3) Environmental Microbiology: The study of how microbes interact with the
environment and each other, including their effects on the landscape, the spread
of viruses and bacteria, the distribution of algae, fungi and parasitical organisms

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 and the associated implications for human health and the environment.
Environmental microbiology also researches how microbes can be used to solve
global problems.

(4) Industrial Microbiology: This is a branch of applied microbiology in which

microorganisms are used in industrial processes; for example, in the production
of high-value products such as drugs, chemicals, fuels and electricity.

(5) Pharmaceutical Microbiology: This is a specialist area of microbiology and one

concerned with the use of microorganisms in pharmaceutical development and
with maintaining contamination control.

(6) Agricultural Microbiology: This is a branch of microbiology dealing with plant-

associated microbes and plant and animal diseases.

(7) Veterinary Microbiology: Branch of Microbiology that is concerned with bacterial

and viral diseases of domesticated vertebrate animals (livestock, companion
animals, fur-bearing animals, game, poultry, but excluding fish) that supply food,
other useful products or companionship.

(8) Food and Dairy Microbiology: The scientific study of microorganisms associated
with food, milk and milk products in all aspects and those that are used in the
production of food and dairy.

(9) Microbial Biotechnology: This is the use of microorganisms to obtain an

economically valuable product or activity at a commercial or large scale. The
microorganisms used in industrial processes are natural, laboratory-selected
mutant or genetically engineered strains.

(10) Aeromicrobiology: This is the study of living microbes which are

suspended in the air. These microbes are referred to as bioaerosols.

(11) Vaccinology: This is the science of vaccines, and historically includes

basic science, immunogens, the host immune response, delivery strategies and
technologies, manufacturing, and clinical evaluation.

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(12) Water/Aquatic Microbiology: Aquatic Microbiology is devoted to
advancing the study of microbes in aqueous environments, with a focus on
freshwater, estuarine and oceanic ecosystems. Aquatic microorganisms play
diverse roles in ecosystems, and are key to earth's biogeochemical cycles. The
water microbiology deals with the examination of water which aims at the
detection or quantification of a great variety of organisms including viruses,
bacteriophages, bacteria, fungi, yeasts and protozoa.

(13) Chemotherapy: Antimicrobial chemotherapy involves the administration of

drugs with selective toxicity against pathogens involved in infections, not host
cells. Antibiotics, which are agents used to combat bacteria, are among the most
common antimicrobials.

GENERAL CHARACTERISTICS OF MICROORGANISMS

Microorganisms as stated earlier, include the protozoa, bacteria, blue-green algae,

fungi and viruses. They are generally called protista. The protista is divided into
simple and complex. The simple protists are the fungi, bacteria and viruses while
the complex protists are protozoa and the blue-green algae. The reason for this
classification is that the simple protists are primitive and unicellular while the
complex ones are multicellular.

Based on biochemical and structural analysis, Neil and Nester discovered that
there were two lines of evolutionary sequence of microorganisms on this basis.
They classified microbes as either prokaryotes or eukaryotes. They considered
as evidence many internal structures as described below:

Nucleus: Eukaryote members have their nucleus bounded by nuclear membrane

while prokaryotes have no nuclear membrane.

Mitochondria: Present in Eukaryotes but absent in prokaryotes

Chromosomes: Eukaryotes have many while prokaryotes have only one

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Chloroplast: This is only in eukaryotic microbes but absent in the prokaryotic
ones

Vacuole: This is also present in eukaryotes but absent in prokaryotes

Endoplasmic reticulum: Also present in eukaryotes but in prokaryotes.

General Characteristics of Microorganisms

(1) Microorganisms generally reproduce asexually by binary fission, budding,

fragmentation and septation.

(2) Microorganisms are unicellular, they exist in single cell but fewer exist in
colony such as volvox. Although viruses are acellular and are termed as
obligate parasites, they cause diseases of both plants and animals and also
bacteria.

(3) Microorganisms are cosmopolitan. They can be found in almost all parts of
the world, ranging from temperate environments to hot springs and Antarctic
regions of the world.

(4) Some microbes possess photosynthetic pigments such as xanthophyll and

carotene while the non-pigmented ones exhibit heterotrophic form of nutrition
by either one of the parasitism, commensalism, amensalism, symbiosis,
saprophytism etc. Some bacteria do not carry out any of the above type of
nutrition but absorb nitrogen from the atmosphere, e.g. Nitrobacter spp.

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(5) Some microorganisms possess rigid cell walls surrounding the cell
membrane while others have flexible cell walls.

(6) They possess genetic material in the form of DNA or RNA

Reproduction in Microorganisms

Microorganisms generally reproduce asexually under favourable conditions.

However, they reproduce sexually when the conditions are unfavourable.

Reproduction involves three aspects thus;

1. Cell division

2. Macromolecular synthesis

3. Growth

The cell division can further be divided into fission, fragmentation, septation, and
budding

The various types of microbial reproduction are described below.

Transverse Binary Fission

This is an asexual form of reproduction whereby a single cell divides into two
after the formation of a transverse septum (cross wall). The overall process is
initiated by the macromolecular synthesis. This is where the bacterial cell
selectively takes up nutrients from the environment and converts them into
macromolecules such as RNA, DNA, protein, enzymes etc. Cell mass and cell

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size increase and are the cell is prevented from bursting by turgor pressure. New
cell building blocks are synthesized. After macromolecules synthesis the DNA
replication follows. Then follows the inward growth of cytoplasmic membrane
and cell wall to produce a septum that ultimately separates the cells and finally
the cell separates into two daughter cells.

Bacterial binary fission

Budding

Budding is a type of asexual reproduction where the new organism (offspring)

grows as an outgrowth from the body of the parent. Here, the new individual
starts growing as a small body on one side of the parent organism and continues
growing in size while still attached to the parent. Early on, it appears as part of
the parent given that it does not detach until it has grown further. Ultimately, the
new individual, which resembles the parent, detaches and becomes an
independent organism.
Various types of bacteria exhibit budding as a mean for reproduction such as
Hypomicrobium spp., Rhodopseudomonas acidophila, Planctomyces spp., some
cynobacteria such as Microcystis aeruginosa, Anabaena circinalis etc. Several

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species of yeasts also exhibit budding such as Saccharomyces cerevisiae,
Candida albicans etc. Some species of Hydra such as H. vulgaris, H. oligactis, H.
Canadensis etc. also exhibit budding as their means of reproduction.

Budding in Bacteria

Budding in Yeast

Fragmentation

This is a form of reproduction where a new organism grows from a fragment of

parent cell. It involves the splitting of microbial filament into smaller filaments
such as bacterial coccal or bacillary filaments each of which gives rise to a new

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growth. It is commonly found in bacteria that produce extensive filament such as
Norcadia spp.

Fragmentation in Bacteria

Spore Formation

The genus Streptomyces and other related bacteria produce many spores by
developing cross walls (septa) at their hyphal tips. Each spore gives rise to a new
organism. The spore may either be a conidiospore or sporangiospore.

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Spore formation in microorganisms

Septation

This occurs in Spirogyra. When this organism reaches maturity, it produces

another cell which becomes separated from the parent cell by the use of a
septum and later the septum separates from the parent organism to produce a
new one.

Septation in Spirogyra

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 MICROBIAL GROWTH

When bacteria are inoculated into a suitable medium and incubated under
appropriate and suitable condition, a tremendous increase in the number of cells
occurs within a relatively short time. In some species, the maximum population is
reached within 24 hours but others require a much longer period of incubation to
reach the maximum growth. The term growth as applied to microorganisms
usually refers to the increase in the total microbial population rather than
increase in the size or mass of an individual organism as it applies to higher
plants and animals. More frequently than not, the inoculum contains thousands
of organisms.

Growth in MCOs denotes the increase in number beyond that which is present in
the original inoculum. Therefore determination of growth requires quantitative
measurement of total population of the cell or cell crops at the time of
inoculation and the gain after inoculation. The most common means of bacterial
reproduction is binary fission, one cell divides; producing two cells, thus if we
start with a single cell bacterium, the increase in population (growth) is by
geometrical progression.

→ →
1−−−−− 21−−−−− 22−−−−− 23−−−−− 2n → →

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The mathematics of population growth (a) Starting with a single cell, if each product of reproduction goes on
to divide in a binary fashion, the population doubles with each new division cycle or generation. This process can
be represented by logarithms (2 raised to an exponent) or simple numbers. (b) Plotting the logarithm of the cells
produces a straight line indicative of exponential growth, whereas plotting the cell numbers arithmetically gives a
curved slope.

Where n = the number of generation. For each succeeding generation, assuming there is
no cell death, doubles the population. The total population N at the end of a given period
would be expressed as

N = 1 x 2n…………………………equation 1

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Microbial growth assumption of one cell as the starting population

However, under practical condition, the number of bacteria (No) inoculated at the time
period is not one but more likely, several thousands. So the Log formula now becomes

N = N0 x 2n……………………………………………………..equation 2

Now solving the equation 2 for n

Multiply both sides by Log in equation 2

Log N = Log N0 x 2n

Log N = Log N0 + Log 2n

Log N = Log N0 + n Log 2

Log N - Log N0 = n Log 2

Log N - Log N0 = n
Log 2

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n = Log N - Log N0. Therefore n = 3.3 (Log N - Log No)…………number of generation
0.301
Thus, by the use of above equation we can calculate the number of generations that
have taken place provided we know the initial population and the population after
growth has occurred.

The rate of growth during the exponential phase in a batch culture can be expressed in
terms of the mean growth rate constant (k). This is the number of generations per unit
time, often expressed as the generations per hour.

k=n = Log N - Log N0

t 0.301t

The time it takes a population to double in size—that is, the mean generation time or
mean doubling time (g), can now be calculated. If the population doubles (t =g), then

N= 2 N0.
Substitute 2N0 into the mean growth rate equation and solve for k.

k = Log (2N0) - Log N0 = Log 2 + LogN0 - Log N0

0.301g 0.301g

k=1
g
The mean generation time is the reciprocal of the mean growth rate constant.

g=1
k

Normal Microbial Growth Curve

Assume that a single bacterium has been inoculated into a plate of liquid culture
medium (broth) which is subsequently incubated. Eventually the bacterium will undergo
binary fission and a period of rapid growth will ensue in which the cell multiplies at an
exponential rate during the period of rapid growth. If we use the theoretical number of
bacteria which should be present at various intervals of time and the data is plotted as
the logarithm of number of bacterial cells versus time, we would obtain a curve as

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follows;

From the above graph, it can be seen that there is an initial period of what appears to be
no growth (Lag phase), followed by rapid growth (the Exponential or Logarithmic phase),
then the leveling up (the Stationary phase) and finally a decline in the viable (capable of
surviving) population (Death or Decline phase). Between each of the phases, there is a
transitional period (curve portion); this represents the time required before all the cells
in a population can enter the new phase.

Lag Phase

The lag phase is a relatively “flat” period on the graph when the population appears not
to be growing or is growing at less than the exponential rate. Growth lags primarily
because: (1) The newly inoculated cells require a period of adjustment, enlargement,
and synthesis; (2) the cells are not yet multiplying at their maximum rate; and (3) the

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population of cells is so sparse or dilute that the sampling misses them. The length of
the lag period varies somewhat from one population to another. In this stage, the
population remains temporarily unchanged. But this does not that the cells are
quiescent or dormant. On the contrary during this stage the cells carry out the following:
(1) Increase in size beyond their normal dimension.
(2) Synthesis of new protoplasm i.e. they are physiologically active.
(3) Synthesis of enzymes and coenzymes to the required amount for optimum
operation of chemical machinery of the cell if deficient.
(4) The organisms are metabolizing but there is lag in the cell division.
(5) Time for adjustment in the physical environment around cell may be required.

At the end of this phase, each organism divides. However, since not all organisms
complete the lag phase simultaneously, gradual increase in population until the end of
the period when all the cells are capable of dividing at regular interval.

Log (Exponential) Phase

When the bacteria have acclimatized to their new environment and synthesized the
enzymes needed to utilize the available substrates, they are able to start regular division
by binary fission. This leads to the exponential increase in numbers referred to above.
Under optimal conditions, the population of cells will double in a constant and
predictable length of time, known as the generation (doubling) time. The value for the
widely used laboratory bacterium E. coli is 20 min, and for most organisms it is less
than an hour. There are some bacteria however; whose generation time is many hours.

During this stage, the cell divides steadily at a constant rate, and the log of number of
cells plotted against time resulting in a straight line. In this phase;
1. There is no competition for nutrient because the nutrients are in excess
2. There is no accumulation of toxic metabolites
3. Suitable temperature, condition and also rapid multiplication

Note that not all bacteria have the same generation time, for some, it may be 15 to 20
minutes. For others, it may be many hours as in the table below
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Bacteria Growth Medium Temperature Generation Time
o
Escherichia coli Milk 37 C 12.5 minutes
Bacillus thermophilus Broth 37 oC 17 minutes
Mycobacterium Synthetic 37 oC 792-932
tuberculosis
Staphylococcus aureus Broth 37 oC 27
Lactobacillus acidophilus Milk 37 oC 66-67
Bradyrhizobium japonicum Salt + Yeast 25 oC 344-469
Treponema pallidum Rabbit Testes 37 oC 1,980

Stationary Phase

This is where the growth eventually decline and stops. The cessation of growth can be
attributed to a variety of circumstances particularly the exhaustion of nutrients and less
often the production of toxic products during growth. The population at the stationary
phase remains constant for a time perhaps as a result of complete cessation of growth
(division) or perhaps because the production rate is balanced by an equivalent death
rate.

Decline (Death) Phase

Following the stationary phase, the microorganisms may die faster than new cells are
produced, if indeed some cells are still reproducing. The most important conditions that
contribute to the death of the microbial cells are as follows:
(1) Depletion of essential nutrients
(2) Accumulation of inhibitory products such as acid
The death rate of microbes at this phase is highly variable, being dependent on the
environment as well as nature of the organisms.

Microbial Growth Media

Except for certain ecological studies where bacterial populations are examined in their
natural habitat, bacteria are usually cultivated and studied under laboratory condition.

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Numerous media have been developed for bacterial cultivation. Because the nutritional
requirements of bacteria vary widely, there are great differences in chemical
composition of the media used in the laboratory. Bacteria also exhibit wide differences
of with respect to the physical condition favouring their growth such as temperature, pH
and gaseous environment. The successful cultivation of bacteria requires an awareness
of all of these factors.

Types of Media

1. General Purpose Growth Media (GPGM)

These are growth media that support the growth of almost all type of microbes
especially all forms of bacteria and fungi. Example include Nutrient agar (NA),
Nutrient broth (NB), Mueller Hinton agar (MHA), Tryptic soy agar (TSA) and
Tryptic soy broth (TSB), Saboraud Dextrose agar (SDA) etc.

2. Selective media

These type of media provide the nutrient that ensure the growth and
predominance of a particular kind of bacterium and do not enhance (and may
even inhibit) other types of organisms that may be present. For example the
isolation of Neisseria gonorrhoea the agent of gonorrhoea from a clinical
specimen is facilitated by the use of media containing certain antibiotics. Those
antibiotics do not affect the N. gonorrhoea but inhibit the growth of the
contaminating bacteria. Example of selective media includes MacConkey agar
(MA), Mannitol salt agar (MSA), Salmonella-Shigella agar (SSA), Eosin
Methylene Blue (EMB) agar etc.

3. Enriched Media

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 These are mainly general purpose media on which some additives are added to
support the growth of other organisms that need special requirements. Example
includes Blood agar that supports the growth of alpha and beta haemolytic
streptococci, chocolate agar etc.

4. Enrichment Media

These are media prepared in the industry purposely to support some organisms
to overgrow others upon cultivation before selective isolation. In these types of
media, the organisms under investigation grow faster than the unwanted bacteria
so as to be identified for selective isolation. Examples include alkaline peptone
water for the enrichment of Vibrio cholerae, Rappaport Vassiliadis broth for the
enrichment of non-typhoidal Salmonellae.

Differential Media

These are media incorporated with certain dyes or reagents which allows
differentiations of various kinds of bacteria. Example, if a mixture of bacteria is
inoculated into a blood containing medium (blood agar), some bacteria
haemolyze (destroy) the red blood cell while others do not. These can be
distinguished as haemolytic and non-haemolytic bacteria. They are mostly
identified by the colour change in the colonial appearance Example include
MacConkey agar, EMB agar, MSA etc., which are also selective media. Thus, a
medium can be both differential and selective for bacterial identification.

Nutritional Requirement and Nutritional Type of Bacteria

All forms of life from microbes to humans share certain nutritional requirements

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 for growth and normal functioning. Example, all organisms require the following
factors/nutrients; carbon, energy, electron, nitrogen, oxygen, sulphur, phosphorus,
vitamins, water and metal ions.
Bacteria can be divided into many groups on the basis of their nutritional
requirements. The major separation is into two groups; phototrophs (those
bacteria that depend on light as the source of energy) and chemotrophs (those
bacteria that depend on chemical compounds as the source of energy). The
phototrophs are further divided into photolithotrophs (i.e. those bacteria that
depend on light as the source of energy and inorganic compounds as source of
electrons such as Chromatium okeinii) and photoorganotrophs (bacteria that
depend on light as energy source and use organic compounds such as Succinate
as a source of electrons; e.g. Rhodospirillum spp.).

Chemolithotrophs on the other hand are those bacteria that obtain their energy
from reduced inorganic compounds such as ammonia (NH3) as electron source.
Example include Nitrosomonas spp.. The Chemoorganotrophs are those
bacteria that obtain their energy from chemical compounds and use organic
compounds such as monosaccharides, amino acids and glucose as electron
source. Example includes Pseudomonas pseudoflora.

One important carbon source that does not supply hydrogen or energy is carbon
dioxide (CO2). This is because CO2 is oxidized and lacks hydrogen. Probably all
microorganisms can fix CO2—that is, reduce it and incorporate it into organic molecules.
However, by definition, only autotrophs can use CO2 as their sole or principal source of
carbon. Many microorganisms are autotrophic, and most of these carry out
photosynthesis and use light as their energy source. Some autotrophs oxidize inorganic
molecules and derive energy from electron transfers.

The reduction of CO2 is a very energy-expensive process. Thus many microorganisms

cannot use CO2 as their sole carbon source but must rely on the presence of more
reduced, complex molecules such as glucose for a supply of carbon. Organisms that

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use reduced, preformed organic molecules as carbon sources are heterotrophs (these
preformed molecules normally come from other organisms). Most heterotrophs use
reduced organic compounds as sources of both carbon and energy.

There are also fastidious heterotrophs which are organisms that have elaborate
requirement for specific nutrients. This means that the organisms have very
complicated nutritional requirements and will not grow without specific factors present
or in specific conditions. Examples include Staphylococcus aureus, Mycoplasma spp.,
Chlamydia spp., Bacillus subtilis etc.

Obligate parasites on the other hand are those microbes that can only be propagated in
association with a living host. Example includes all viruses, Mycobacterium leprae etc.

Microbial Physical Growth Requirements

It is necessary to know the physical environment in which bacteria grows best in

addition to proper nutrients for their cultivation. Just as bacteria vary greatly in their
nutritional requirements, so do they exhibit diverse responses to physical conditions
such as temperature, pH, and gaseous conditions.

Temperature

This is the degree of hotness or coldness of an environment. The temperature

profoundly influences the pattern of microbial growth. This is because the processes of
growth are dependent on chemical reactions which are in turn influenced by
temperature. On the basis of temperature requirement, microorganisms are categorized
as follows:

Psychrophiles
These are microbes that grow at temperatures ranging from 0 oC to 20 oC with an
optimum temperature of 15 oC. They are described as cold survivors. The term

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psychrophiles or facultative psychrophiles is used to describe bacteria or microbes that
o o o
are able to grow at 0 C but having optimum growth temperature of 20 C to 30 C. The
physiological factors responsible for low temperature maxima for strict psychrophiles
are not entirely clear but some factors that have been implicated are heat instability,
ribosomes and various enzymes, increased leakage of cell components and impaired
transport of nutrient above the maximum temperature.

Mesophiles
These are microbes having optimum growth temperature of 37 oC and within a
temperature range of 25 oC to 40 oC. For example, all pathogenic (disease-causing)
bacteria for human and worm blooded animals are mesophiles.

Thermophiles
These are microbes that can grow at temperature of 45 oC. The growth of many
thermophiles extends to mesophiles region and the organisms are designated
facultative thermophiles. Other thermophiles cannot grow in mesophilic range and
these are termed obligate thermophiles. Example includes Stenothermophilus spp.

Gaseous Requirement

The principal gases that affect microbial growth are oxygen and carbon dioxide.
Bacteria display a wide variety of responses to free oxygen that is convenient to divide
them into four groups on the following basis:

Aerobic Bacteria
These are bacteria that can survive in an oxygenated environment. They require oxygen
for growth and can grow when incubated in an air atmosphere (i.e. 21% O2). Example
includes Mycobacterium tuberculosis, Pseudomonas aeruginosa, Bacillus spp.,
Nocardia asteroids etc.

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Anaerobic Bacteria
These bacteria do not use oxygen to obtain energy. Oxygen is toxic for them and they
cannot grow when incubated in air atmosphere. Some can tolerate low level of oxygen
and they are termed as non-stringent or tolerant anaerobes, but others cannot tolerate
even low levels and may die upon brief exposure to air. Examples include Clostridium
tetani, Bacteroides spp., Propionibacterium spp., Fusobacterium spp., etc.

Facultative Anaerobic Bacteria

This group of bacteria does not require oxygen for growth although they may use it for
energy production if it is available. They are not inhibited by O2 and usually grow as well
under an air atmosphere as they do in the absence of oxygen. Examples include
Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, Yersinia pestis,
Morganella morganii, Salmonella spp. etc.

Microaerophilic Bacteria
These bacteria are those that require very low levels of oxygen for growth but cannot
tolerate the level of oxygen in the atmosphere. Examples include Helicobacter pylori
(present in ulcer patients) and Borrelia burgdoferi (the agent of Lyme disease).

pH Requirement

pH is the degree of acidity or alkalinity of a medium. It is expressed by the pH scale, a

series of numbers ranging from 0 to 14. The pH of pure water (7.0) is neutral, neither
acidic nor basic. As the pH value decreases toward 0, the acidity increases, and as the
pH increases toward 14, the alkalinity increases. The majority of organisms lives or
grows in habitats between pH 6 and 8 because strong acids and bases can be highly
damaging to enzymes and other cellular substances. On the basis of pH, bacteria are
classified into three as;

1. Acidophiles (Acidophilic Bacteria)

These are bacteria that prefer to grow in an approximately acidic pH (i.e. less
than 6.5). Example includes Lactobacillus spp., Acidobacterium capsulatum,
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 Thiobacillus acidophilus, etc.
2. Alkalophiles/Basophile (Alkalophilic/Basophilic Bacteria)
These are bacteria that prefer to grow in an extremely basic or alkaline
environment. They grow in pH above 7.5. Examples include Vibrio cholerae,
Micrococcus spp., Pseudomonas spp. etc.
3. Neutrophiles
These are bacteria that prefer to grow in an approximately neutral pH. Examples
include Staphylococcus aureus, Shigella spp., Salmonella spp., E. coli, etc. Most
of pathogens (the disease causing organisms) are neutrophiles.

Whatever the preference of microbes at the onset of culture, microorganisms have the
ability to alter the pH of their local environment during the culture process. They do this
by making either acid or alkaline product. Example is the ulcer bacterium the
Helicobacter pylori if found in human stomach, a highly acidic environment. It can
survive because it elaborates a highly efficient urease that splits urea to ammonia
which is alkaline. This raises the pH of the microenvironment thereby preventing the
bacterium from acid attack via neutralization reaction.

MICROORGANISMS AS FRIENDS AS FOES

The positive and negative impacts of microorganisms are inestimable. Microbes act as
friends in which their presence benefit humans and as well act as foes (enemies) in
which they harm humans in many ways. Microorganisms’ involvement in various
processes that have impact on human lives as
(a) Fermentation process
(b) Cause of infection and diseases
(c) Microbes in food industries
(d) Production of antibiotics
(e) Deteriorating agents
(f) Involvement of microbes in sewage and water treatment
(g) Involvement of microbes in nutrient recycling
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 (h) Microbes in the petroleum industries

Role of microorganisms in Fermentation

Fermentation is the anaerobic breakdown of carbohydrates by anaerobic

microorganisms which derive their energy from complex organic substances. The end
product of fermentation is alcohol e.g. ethanol, which is used in the production of other
compounds.

In the production of alcohol, complex carbohydrates such as corn, grape, molasses,

sugar beets, potatoes are used as the raw materials. It is first necessary to hydrolyze
them into simple fermentable sugars. This hydrolysis is accomplished with enzymes
from microorganisms called yeast, mold or by heat treatment of acidified materials.
Certain chemicals are added to inhibit the growth of microorganisms. The microbe that
carbohydrate grows naturally on the carbohydrate, but if the growth is insufficient to
accomplish the fermentation; a culture of organism called starter culture is added. An
example of the microorganism involved is Saccharomyces cerevisiae. It is imperative
that the culture be one that grows vigorously and has a high tolerance for alcohol as
well as capacity for producing a large yield of alcohol.

C6H12O6 2C2H5OH + 2CO2

However, production of alcohol is not only limited to the use of yeast alone, other
bacteria such as Zymomonas mobilis ferment carbohydrate directly to ethanol and the
speed of the reaction is twice than that of yeast. Moreover, Thermoanaerobacter
ethalicus is very efficient in the formation of alcohol from carbohydrates. The alcohol
produced in the above process is used in many industries as solvent and for the
preparation of other chemicals, for example, alcohol is used as a solvent in paint
industries and as a substance in the production of acetic acid. This involves the
alcoholic fermentation of carbohydrates to alcohol and consequently oxidation of
alcohol to acetic acid.

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2C2H5OH + 2O2 2CH3COOH + 2HO2
Acetic acid is used as preservative in photographic processing

Uses of Microbes in Food Industries

Microbes perform important function in the food industries. They bring about changes
and also speed the rate of chemical processes involved in making some food products.
For example butter, cheese, and yoghurt are the products of milk fermentation. Lactic
acid can also be produced by breaking down of lactose in the presence of an enzyme,
lactase produced by certain microbes. The bacteria used in lactic acid fermentation
include Lactobacillus bulgaricus, Lactobacillus helveticus and Leuconostoc spp. Lactic
acid is used as preservative in the textile industries for treating fibres. It is also used in
electroplating as copper lactate and as baking powder as calcium lactate. Other
fermented products with the microbes responsible for their production include:
1. Cultured buttermilk------ by a mixture of lactic acid streptococci (Streptococcus
lactis, S. cremoris) with aroma-producing bacteria (Leuconostoc citrovorum or L.
dextranicum)
2. Yoghurt-------- Streptococcus thermophilus, Lactobacillus bulgaricus
3. Cheese------- Streptococcus lactis, S. cremoris, Leuconostoc cremoris,
Lactobacillus helveticus, Streptococcus thermophilus.
Microorganisms are also employed as colouring substances; for example, Penicillium
oxalicum. Some yeasts are also employed in brewing industries for the production of
alcoholic beverages such as:
1. Beer----------------Saccharomyces cerevisiae or Saccharomyces carlsbergensis
2. Whiskey----------- Saccharomyces cerevisiae
3. Wine--------------- Saccharomyces ellipsoideus.
Some microbes are also involved in food spoilage such as;
1. Alteromonas spp., Shewanella putrefaciens, Erwinia carotovora for the spoilage
of cold stored foods such as dairy, vegetables, fish, poultry and meat.
2. Bacillus stearothermophilus and Clostridium thermosaccharolyticum for the
spoilage of canned foods

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 3. Rhizopus nigricans, Penicillium spp. and Aspergillus spp. for the spoilage of
vegetables and bakery foods

Microbes in the Production of Antibiotics

Antibiotics are products of metabolisms of some microbes that inhibit or kill other
microbial life forms. They are extensively used in chemotherapy for curing many
diseases cause by microorganisms. The first scientist to discover the production of
antibiotics is Alexander Fleming (1927) and it was not recognized ever since until after
the Second World War. Antibiotics are grouped into two broad divisions;
(1) Those that are specific in their action; and
(2) Those that are general in their activity
Specific antibiotics are used in neutralizing a particular disease only, therefore, they act
on the type of pathogen only and they are also called narrow spectrum antibiotics.
Penicillin is a narrow spectrum antibiotic that is produced by Penicillium chrysogenum.
Those antibiotics that are general in their action are also known as broad spectrum
antibiotics. These include Streptomycin, Chloramphenicol, and Tetracycline. They can
act on both Gram positive and Gram negative bacteria. To produce these antibiotics in
large quantity, the organisms producing them are kept in a fermentative process to
continue metabolizing and thus continue producing antibiotics. Now, when the
metabolic activities have reached their peak, chemical and biological assays such as
chromatography and precipitation are used to separate the antibiotics. Some antibiotics
and the microbes responsible for their production include;
1. Chloramphenicol -----------Streptomyces venezualae
2. Streptomycin----------------Streptomyces griseus
3. Erythromycin---------------- Saccharopolyspora erythraea (formerly known as
Streptomyces erythraeus).
4. Tetracycline------------- Streptomyces aureofaciens
5. Grieofulvin (Fulcin)---- Penicillium patulum

Microorganisms in Causing Infection and Disease

Some microorganisms are used to cause diseases to humans, animals or both. The

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degree of infection depends on the microbial population growth within the host, time
taken and the presence of some virulent factors in the host. Examples are tabulated
below
S/N Microorganism Type of Microbe Disease it causes
1. Salmonella typhi Bacterium Typhoid fever
2. Mycobacterium tuberculosis Bacterium Tuberculosis
3. Mycobacterium leprae Bacterium Leprosy
4. Staphylococcus aureus Bacterium Boils, carbuncle, food
poisoning
5. Vibrio cholera Bacterium Cholera
6. Shigella dysenteriae Bacterium Dysentery
7. Neisseria meningitidis Bacterium Cerebrospinal meningitis
8. Clostridium tetani Bacterium Tetanus
9. Candida albicans Yeast (Fungus) Vaginal and oral thrush
10. Microsporum canis Mold (Fungus) Ringworm of the head
11. Malassezia globosa Mold (Fungus) Dandruff
12. Morbillivirus Virus Measles
13. Human Immunodeficiency Virus AIDS
virus
14. Bordetall pertusis Bacterium Whooping cough
15. Human Ebola Virus Virus Ebola haemorrhagic fever

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