MICROBIOLOGY

HNR 2102

WEEK 1; INTRODUCTION TO MICROBIOLOGY

 Modern Developments in Microbiology
Bacteriology is the study of bacteria.
Mycology is the study of fungi.
Parasitology is the study of protozoa and parasitic worms.
Virology is the study of Viruses
Recent advances in genomics, the study of an organism’s genes, have
provided new tools for classifying microorganisms.

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 Modern Developments in Microbiology
Recombinant DNA is DNA made from two different sources. In the 1960s,
Paul Berg inserted animal DNA into bacterial DNA and the bacteria
produced an animal protein.
Recombinant DNA technology, or genetic engineering, involves
microbial genetics and molecular biology.

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 Structure of Bacteria
Microscopic, simple, living
Prokaryotic, no true nucleus
Non-membrane bound organelles
More complex than viruses but less than living, cellular organisms
Ribosomes smaller
Inherited information held in single circular chromosome

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5
 Classification of Bacteria
Shape
·
Spheres
⑲ coccus

o
Rods
bacillus

⑧
Spiral
spirillum

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 Cell Arrangement

⑦
Diplo – cells are paired

④ ⑥
Staphylo – cells are in grape-like clusters

⑥
Strepto – cells are in long chains

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 Differences between Bacterial and Human Cells
• Bacterial cells
Human cells N
v Nucleus
• No nucleus
↑ • No intracellular organelles (but ribosomes)
r Intracellular organelles
(ribosomes subtly different) v • No introns (nearly)
↑ p • No junk DNA
Introns
r • Plasmids, bacteriophage
U Lots of junk DNA
⑤
Viruses

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 Bacterial Reproduction
Prokaryotes reproduce asexually by binar fission thus
making new DNA almost continually .
They can exchange genetic material in three ways:

↓- transformation where genes are taken up from the

surrounding environment,

2- conjugation where genes are transferred from cell to

cell and
②- transduction where genes are transferred between
3/ prokaryotes by viruses

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 Bacterial division by binary fission
Im
2

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 Fungi

Eukaryotes

Chitin cell walls

Use organic chemicals for energy.
Molds and mushrooms are multicellular consisting of masses of mycelia,
mmm
which are composed of filaments called hyphae.
Yeasts are unicellular.

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Figure 1.1b
 Protozoa
Eukaryotes
Absorb or ingest
organic chemicals
May be motile via pseudopods, cilia,
or flagella

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Figure 1.1c
 Viruses
Acellular
Consist of DNA or RNA core
Core is surrounded by a protein coat.
Coat may be enclosed in a lipid envelope.
Viruses are replicated only when they are in a living host cell.

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Figure 1.1e
 Microbes and Human Disease
Bacteria were once classified as plants giving rise to use of the term flora
for microbes.
This term has been replaced by microbiota.
Microbes normally present in and on the human body are called normal
microbiota or commensals
Saprophytes
Opportunists

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 The oral cavity “the most dirty part in our body”

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 Normal Microbiota
Normal microbiota prevent growth of pathogens.
Normal microbiota produce growth factors such as folic acid and vitamin K.
Resistance is the ability of the body to ward off disease.
Resistance factors include skin, stomach acid, and antimicrobial chemicals.

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 Infectious Diseases
When a pathogen overcomes the host’s resistance, disease results.

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 Not all bacteria are bad!!

• Many bacteria aid in the decomposition dead

organisms and waste.
• Bacteria control the nitrogen cycle.
• Bacteria is used to make cheese and yogurt.
• Bacteria can be used to make antibiotics.
• Bacteria are used to make sauerkraut.
• Bacteria are used to clean up oil spills.
• Bacteria are very useful in genetic engineering.

https://www.youtube.com/watch?v=ycO-
oWYvaQI&t=22s
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 Observing microorganisms
Culture Media technique
Media is a substance used for growing bacteria and other
microorganisms in the laboratory.
May be liquid media or solid media
- Broth is an example of liquid media
Agar plates are an example of solid media.
-
In a laboratory media is used to help the growth, isolation and
identification of microorganisms. mm
ma

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 Types of Specimens
W• Respiratory Throat swab & sputum
• Fluids
W• Wounds
Cerebrospinal fluid
Surgical or nonsurgical
• Genital

Y•
•
Urine
Stool
High vaginal swab
Clean catch or catheterized
⑤• Eye Stool
5•
•
Ear
Skin
Swab
Swab
Swab
• https://www.youtube.com/watch?v=cneascR3OEc

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 Visualization of microbes
A microscope is an optical instrument that uses a lens or a combination of lenses
to magnify and resolve the fine details of an object.

The magnified image seen by looking through a lens is known as a virtual

image, whereas an image viewed directly is known as a real image.

The object to be magnified is placed under the lower lens, called the objective
and viewed through the upper lens, called the eyepiece.

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 Microscope
Microscope is the instrument used to see very small
things which cannot be seen by the naked eye.
Most microbes are so small that they are measured in
micrometers or nanometers.
A typical bacterial cell is about 1 um while a virus is
more in the range of 10-100 nm.

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 Types of Light Microscope

* The bright-field microscope produces a dark image against a brighter

background
The dark-field microscope produces a bright image of the object against
* a dark background and is used to observe living, unstained preparations

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 Types of Light Microscope
The phase-contrast microscope enhances the contrast between intracellular structures
↑ that have slight differences in refractive index and is an excellent way to observe living
cells

↑
The fluorescence
mn
microscope exposes a specimen to ultraviolet, violet, or blue light and
shows a bright image of the object resulting from the fluorescent light emitted by the
specimen

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 How does microscope work?

0
1. produce a magnified image of the specimen
2. separate the details in the image,
3. render the details visible to the human eye or camera.

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 Scale

1 micrometer = 1/1000000 meter

(six zeros) = 106

1 nanometer = 1/1,000,000,000
(nine zeros) = 109
(one one-billionth) of a meter

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 Microscope

One or more lenses that make an enlarged image of an

object.

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 Simple Microscope
Light passes through only 1 lens.
Example: magnifying glass

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 Compound Microscope
Lets light pass through an object and then through two or more lenses.

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 Electron microscopes
• use a beam
mmuno
of electrons instead of a beam of light to
magnify the image

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 The Scanning Electron Microscope
produces a 3-dimensional image of specimen’s surface features

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 Transmission electron microscopy
(TEM).
• Allows the observation of molecules inside the
cells

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 https://www.youtube.com/watch?v=SUo2fHZaZCU

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 U n Place the Slide on the
Microscope
v n Use Stage Clips

N n Click Nosepiece to the

lowest (shortest) setting
n n Look into the Eyepiece
~n Use the Coarse Focus

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 Identification
his
Gram Stain

1 3
The Gram stain differentiates bacteria on the basis of
structure and composition of the layers of the cell wall.
Upon completing the stain, Gram positive bacteria
appear purple and Gram negative bacteria appear pink.
Gram positive cell walls have a simpler structure than
Gram negative cell walls. Also note the shapes of
bacteria represented here

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 Simple Stains
Simple stain: Use of a single basic dye e.g methylene blue
A mordant may be used to hold/fix the stain or coat the specimen to
enlarge it.

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 Differential Stains: Gram Stain
The Gram stain classifies bacteria into gram-positive
or gram-negative. ⑰
⑥ Gram-positive bacteria tend to be killed by penicillin and detergents.
⑦
· Gram-negative bacteria are more resistant to antibiotics.

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 Differential Stains: Gram Stain

Color of Color of
Gram–positive Gram–negative
cells cells

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Primary stain: Purple Purple
Crystal violet
Mordant: Purple Purple
Iodine
Decolorizing agent: Purple Colorless
Alcohol-acetone
Counterstain: Purple Red
Safranin
PLAY Animation: Microscopy
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 Differential Stains: Gram Stain

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Figure 3.11b
 Differential Stains: Acid-Fast Stain

Cells that retain a basic stain in the presence of acid-alcohol are called acid-fast.

Non–acid-fast cells lose the basic stain when rinsed with acid-alcohol, and are
usually counterstained (with a different color basic stain) to see them.

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Figure 3.12
 Special Stains
Negative staining is useful for capsules.
Heat is required to drive a stain into endospores.
Flagella staining requires a mordant to make the flagella wide enough to see.

https://www.youtube.com/watch?v=sxa46xKfIOY

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Figure 3.13a–c
 What are the“Basic Skill of microbiology”?

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 HEALTH AND SAFETY

1. Basic Safety Rules

Colleges and workplaces must provide a working environment that is safe
and without risk to health.
Training on safe working practices must be given.
Safety in the clinical facility is EVERY user’s responsibility.
Each person has a duty to keep a safe work environment.
Each clinical facility worker should be able to identify possible dangers
in the working place.

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 Basic Safety Checklist for Laboratory Staff
Did you wash your hands before you left the
hospital?
Did you wear the correct clothing in the
working place (laboratory coat, safety goggles,
gloves and shoes)?

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 Safety signs

There are internationally accepted standard safety and warning signs used in laboratories around the world.
The signs are colour coded into 4 main categories; examples of each are given below:
1- Mandatory
“must do”

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 Safety signs
2) Warning/caution.
These warn of possible dangers.
They are yellow and black.

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 Safety signs
3) Prohibitive: “Don’t do it”
As with the “must do” signs the instruction must be obeyed.

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 Safety signs
Safe condition / information signs.

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 Safety signs
Some signs give combined information and have the relevant signs and
colors.

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