GENERAL MICROBIOLOGY

Definition;
Microbiology is the sciences that deals with the study of microorganism, such as
bacteria, fungi, parasite, virus, algae and their interaction with their
environment.
Microorganism are single cell microscopic organism and viruses which are
microscopic but not cellular.

PATHWAY IN DISCOVERY MICROBIOLOGY

 ROBERT HOOKE (1635 – 1703); an English mathematician and natural
historian was linked to the invention of the microscope.
 ANTONY VAN LEEUWENHOEK (1632 – 1723); was a Dutch. Was the first
person to see bacteria under the microscope.
 FERDINAND COHN (1828 – 1898); was born in Poland and participated in
the invention of the microscope and discover a group of bacteria that
produce endospores.
 A French man LOUIS PASTEUR (1822 – 1895); studied the mechanism of
alcoholic fermentation which is catalyze by living yeast cells.
 ROBERT KOCH (1843 – 1910); discover that bacteria that causes infectious
diseases and can grow in “Miller’s coiled culture media”.

GENERAL CHARACTERISTICS OF MICROORGANISM

1. Some are prokaryotic cells
2. Some are eukaryotic cells
3. Some are acellular agent like virus (does not consist of cell).

SIZE OF A MICROOGANISM
Microorganism varies in size ranging from 10nm(nanometer) to
100nm(nanometer).
1. Virus; it measures 109 m
2. Bacteria; it measures 106 m
3. Parasite (helmets); it measures 103 m

BACTERIOLOGY
Bacteria are a group of single cell microorganism with prokaryotic cellular
configuration. The genetic material of a prokaryote cell exists on bounded
in the cytoplasm of a cell.

STRUCTURE AND FUNCTION OF A PROKARYOTIC

CELL
Prokaryotic cells are protein attached to the cell surface in the form of
flagella and pila, a cell enveloped consisting of a capsule, a cell wall, a
plasma membrane and a cytoplasm region that contain the cells genome,
ribosome and various sort of inclusion.

THE BACTERIA CELL STRUCTURE

1. Flagella; these are filamentous protein structure attached to the cell wall.
Function
  They provide swimming movement for most motile prokaryote cell.
The distribution of flagella is;
 Monotrichous; a single flagellum at one end of the cell
 Lophotrichous; a bunch of flagella at one end
 Amphitrichous; flagellum at each end
 Peritrichous; flagella everywhere of the bacteria

2. Pilli; these are hairlike structure on the surface of the bacteria cell. They
are primarily composed of protein. Pilli connects a bacterium to another
of his specie or another bacterium of a different species and build a bridge
between the interior of the cell.
Function
 It enables the transfer of plasmid between the bacteria and the
transfer of DNA between mating bacteria.

3. Cell wall; around the outside of the cell membrane is the bacteria cell
wall. The bacteria cell wall is made up off peptidoglycan. The bacteria cell
wall is the different from the cell wall of plant and fungi which are made
up off cellulose and chitin.
 Function
 The cell wall gives a shape and protects the inner part of the bacteria.
NB; they are broadly speaking of two different types of cell wall in bacteria;
gram positive and gram negative. The names originate from the reaction of the
cells to the gram stain (colour). Gram positive bacteria possess a thick cell wall
containing many layers of peptidoglycan and teichoic WHILE gram negative
bacteria have a relatively thin layer of peptidoglycan in their cell wall.

4. Cell membrane (cytoplasmic or plasma membrane); is a biological

membrane that separate the interior of the cell from the outer
environment. It consists of lipid by-layer with embedded proteins.
Function
 It provides mechanical support of the cell and allows the passage of
large molecules.

5. Capsule; is a very large structure of some prokaryotic cells, it is a layer that

lies outside the cell wall of bacteria. It consists of polysaccharide and they
are water soluble and difficult to stain.
Function
 A capsule is considered as virulence factor because it enhances the
ability of bacteria to cause diseases (prevent phagocytosis)

6. Ribosome;
Function
 Involves in protein synthesis

7. Nucleic acid (nucleoid DNA); consisting of chromosome, plasmid

STRUCTURE OF THE EUKARYOTIC CELL

 DIFFERENCE BETWEEN PROKARYOTES AND
EUKARYOTES CELLS
SERIES Prokaryotes Eukaryotes
1 Lack a cell nucleus Has a cell nucleus
2 Most prokaryotic are It is unicellular or
unicellular, e.g., multicellular, e.g.,
amoeba animal cell
3 Smaller size of Larger size of
ribosome ribosome
4 It does not have a It does have a
nucleus, nucleus,
mitochondria, and mitochondria and
chloroplast chloroplast
5 Lack nucleic Has a nucleic
membrane membrane
 CLASSIFICATION OF BACTERIA
1. Morphological and arrangement
2. Chemical composition (staining technique)
3. Cultural characteristics
4. Biochemical reaction

MORPHOLOGICAL AND ARRANGEMENT

Bacteria are morphological identified by;
a) Cocci (coccus); they are round or oval in shape. Some of these bacteria
are arranged in pairs (diplococci), chain and cluster.

Coccus diplococci e.g., Neisseria species

Cocci in chain e.g., streptococcus species

Cocci in cluster e.g., staphylococcus species

Diplococci encapsulated e.g., streptococci pneumoniae

b) Bacilli (bacillus); they are rod in shape. They exist in:

Single bacillus e.g., shigella species & salmonella species

Comma shape bacillus e.g., vibrio cholerae
c) Spirochetes; these are spiral bacilli

bacteria that cause syphilis

e.g., leptospiral species

d) Coccobacilli; these are bacteria with half rod and cocci shape.

Short rod (coccobacilli)

e.g., haemophilus influenza & gardenella vaginalis
 CHEMICAL CLASSIFICATION
This is a method of staining the bacteria using dyes. In bacteriology for
the identification of bacteria two main stain are used are used to
classify the bacteria.

a) Gram stain
 Principle of gram stain; it puts in evidence the classification of two
main groups of bacteria, the gram-positive bacteria and the gram-
negative bacteria due to their permeability of the cell made up of
peptidoglycan where the gram positive are stain purple and the gram
negative are stained red or pink.

Method
 Reagent; crystal violet (purple colour)
 Lugol iodine (mordant)
 Alcohol
 Dilute carbolfuchsin (red colour)

Procedure; on a smear
 Pour crystal violet and wait for 1min and rinse
 Then add Lugol iodine to fix the purple colour of crystal
violet and wait for 1min and rinse
 Decolourized with alcohol for 10 – 30 seconds and then
rinse to stop the reaction.
 Then add dilute carbolfuchsin and wait for 1min and allow
it to dry

Observation; under the microscope, observe gram positive bacteria

coloured purple and gram-negative bacteria coloured pink o red.
b) Ziehl nelson stain
  Principle; it puts in evidence of acid fast bacilli (AFB) where
concentrated carbolfuchsin is passed under the flame 3 times
which is going to lyse the cell wall of the bacteria made up of
lipoprotein to ease the permeability of the carbolfuchsin and
it’s read on a blue background.

Method;
 Reagent
 Concentrated carbolfuchsin
 Acid – alcohol
 Methylene blue

Procedure; on a smear
 Pour concentrated carbolfuchsin and pass it on the flame
3 time and wait for 5min then rinse
 Decolourized with acid alcohol for 2 – 3min and rinse
 Then add methylene blue for 2min and rinse and allow it
to dry

Observation; under the microscope, observe red bacilli (AFB) on a blue

background.

CULTURAL CHARACTERISTIC
Bacteria culture is a method or technique in the bacteriology bench used to
isolate and confirm the presence of bacteria growing in the culture media in the
form of colony.
 CULTURE MEDIA; this are milieu or environment used to grow bacteria in
the presence of special nutrients
TYPES OF CULTURE MEDIA
 a) Basic/simple culture media;
These are culture media which allows the growth of all pathogens.
They are also used for the preparation of other culture media. E.g.,
nutrient agar, Columbia agar

b) Enriched culture media;

These are media that have been added special ingredient to encourage
the growth of pathogenic or fastidious bacteria. E.g., blood agar,
chocolate agar

c) Enrichment culture media;

These are culture media that boost up the growth of bacteria and
inhibits or discourage the growth of unwanted pathogen. E.g., selenite
F. broth (salmonella growth), alkaline peptone water

d) Selective culture media;

These are media that allows the growth of specific bacteria. E.g.
 Salmonella /shigella
 Tleosulphate citrate
 Bile salt sucrose (allows the growth of fibrous cholerae)
 Mac conkay agar (allows the growth of gram-negative
bacilli)

e) Differential culture media;

These are culture media that has been added an indignation which is
going to differentiate the presence of a bacteria from another of the
same species.
e.g., CLED (cysteine lactose electrolyte deficiency)
Mac conkay agar (helps to differentiate bacteria that
ferment lactose from those that does not ferment lactose).

 Colonies; these are bacteria growing in a group of community. It is

characterized by it’s colour, height and texture
 Bacteria growth; these is the growing of bacteria under a cultural media
within a particular period of time
 Generation time; this is the time taking for a bacterium to divide itself.
This is observed in the bacteria curve growth above.

BACTERIA GROWTH CURVE

Stationary
Exponential phase phase Decline or death
phase phase

Lag phase

 Lag phase; this is the initial phase where bacteria adapt to their new
environment
 Log or exponential phase; rapid cell division occurs, leading to exponential
growth. Bacteria multiply at a constant rate due to abundant nutrients
 Stationary phase; growth rate slows as nutrients becomes limited. The
number of live cells stabilizes as cell production equals cell death
 Decline or death phase; the number of living bacteria begins to decrease
due to scarce of nutrients and waste products start to build up in the
environment.

BIOCHEMICAL REACTION
Biochemical reaction is significant in the identification of bacteria and in
the identification of different bacteria species
 SOME OF DOES BIOCHEMICAL REACTION USED TO
IDENTIFY BACTERIA
1) Catalase test;
 Aim; the catalase test is aimed to distinguished between the
staphylococcus species that produce catalase enzyme from the
streptococcus species that do not produce catalase enzyme.
 Principle; it puts in evidence the presence of bacteria
(staphylococcus spp) that produce catalase enzymes which is
going to react with the h2o2 (hydrogen peroxide) to produce h2
and o2 in the form of air bubbles.

Method;
 Reagent; h2O2
 Sample; colonies growing on a culture media

Procedure;
1 colony growing from a culture medium

1 drop of h2O2
 On a clean slide, put 1 or 2 drops of h2O2 on the slide
 With a plastic wire loop, pick a colony from a culture media and mix it in
the drop of h2O2. Observe for the release of O2 in the form of air bubbles

Result;
 Presence of air bubbles positive test for catalase
 No air bubbles negative test for catalase
That is for staphylococcus species, we can observe that;
  Staphylococcus species are catalase positive
While
 Streptococcus species are catalase negative

2) Coagulase test;
 Aim; this test is aim to differentiate staphylococcus from the other two
species of staphylococcus.
 Principle; it put in evidence the presence of coagulase enzyme produce by
staphylococcus aureus in the presence of plasma to form agglutination
reaction.

Method;
 Reagent; plasma
 Sample; colonies growth

Procedure;

Colonies suspension

1 drop of plasma
 Put 1 drop of plasma on a slide
 Then add 1 drop of colony suspension and mix
 Observe for agglutination
Result;
 Presence of agglutination coagulase (+) for staphylococcus aureus
 No agglutination coagulase (-) for staphylococcus epidermidis or
staphylococcus saprophyticus.

PARASITOLOGY
Parasitology is the study of parasite
Parasite; an organism that entirely depend depends on another organism called
host.

TYPES OF PARASITES
1) Ectoparasite; parasite that lives on the external surface of a host.
e.g., lies, or arthropods

2) Endoparasite; parasites that lives within the host

e.g., ascaris lumbricoides, entamoeba histolytica, Enterobius vermicularis

3) Obligatory parasite; parasites (organisms) that are completely dependent

on their host for survival and cannot live independently.
e.g., plasmodium species, toxoplasma gondii.

4) Facultative parasite; parasite that does not depend on a host for survival.
e.g., naegleria fowleri, acanthamoeba species

5) Accidental parasite; organisms that parasitize a host other than it’s usual
host
e.g., echinococcus species

6) Erratic parasite; parasites that wander to other organs which is note

usually found
e.g., entamoeba histolytica (in the liver)

HOST
TYPES OF HOSTS
1) Definitive host; the host in which a parasite reaches maturity and
undergoes sexual reproduction.
e.g., in case of plasmodium (the malaria parasite) the definitive host is
the female anopheles mosquito
2) Intermediate host; the host in which the parasite undergoes asexual
reproduction or development but does not reach maturity.
e.g., in the life cycle of toxoplasma gondii, humans can act as
intermediate hosts.

3) Reservoir host; a host that harbours a parasite and can transmit it to

other host, often maintaining the parasite in a population.
e.g., wild animals can serve as reservoir hosts for Lyme disease (carried
by borrelia burgdorferi).

4) Vector; is an agent (insect) that transmit the parasite from one host to
another.
e.g., the female anopheles mosquito

NB: all arthropods are vectors but not all vectors are arthropods.

RELATIONSHIP BETWEEN PARASITE AND VECTOR

1) Symbiosis; it refers to a close, long-term interaction between two
different species. The relationship can be beneficial, harmful or
neutral to one or both parties involved

TYPES OF SYMBIOTIC RELATIONSHIP

 Parasitism; it is a feeding relationship where one
organism called the parasite lives in the other called
the host, live and causes harm to the host.
e.g., taenia saginata.

 Commensalism; it is a relation where the parasite

benefit from the host but does not cause harm (that
parasite is known as commensal)
e.g., entamoeba gingivalis in the mouth.

 Mutualism; a relationship in which both the parasite

and the host benefit from each other.
 e.g., herbivores and the plants they consume, where
herbivores get food and plants benefit from being
eaten (promoting growth).

BASIC CONCEPT IN PARASITOLOGY

In medical parasitology, each of the medically
important parasite are discuss under the standard sub
heading of Morphological, geographical distribution,
means of infection, life cycle, host and parasite
relationship, and clinical manifestation of infection,
laboratory diagnosis, preventions and control
measures.

1) Morphology;
The parasite is classified into two main groups. The first group,
*Protozoa, includes the cyst and the trophozoite (or vegetative form),
with the oocyst serving as a form of identification in various samples.
The second group, **Helminthes* (worms), is characterized by the
identification of the egg, larva, and adult worm.

PROTOZOA
4 nuclei

The cyst of entamoeba histolytica

 pseudopod (false foot)

The trophozoite of entamoeba histolytica