Introductory Lecture:

General Microbiology:
General Microbiology includes physiology, biochemistry, control, growth, morphology and anatomy of
germs.

 Test/procedures take 3 minutes to 1 hour

 Pre-test takes more time (24 hours)
 Pre and post process of practical takes time
 Microbiology is not visible. It is micro.

General sizes of microorganisms:

 Bacteria — 10-6
 Virus — 10-9
 Prions — 10-10
 Fungus — 10-3-6
 Algae — 10-3-6
 These microorganisms are not visible with naked eye.
 Microbiology is a logic based or inference based study.
 Microbes are most enormous in the world.
 Level of microbes is 1012/ml or 1018/ml or 10 Trillion per ml in marshy water.
 Level of microbes in intestine is 1012/ml.
 Microbiology starts from microbiome.
 Microbiome decides the health of humans and animals or microbiome effectively controls the
health and disease status.
 If microbiome is normal, health is normal.
 If microbiome is not normal disease is caused.

SOP's in practicals:
 SOP's stands for "Standard operating procedure/protocol"
 Before each experiment, you should have SOP's.
 SOP's should be noted on practical notebook.

Introduction to microbiology:
Microbiology is the study of microbes.

Biosafety:
"Bio" means "life" and "safety" means "protection".

 Biosafety includes the safety of:

1. Product safety
2. Safety of life

Product safety:
Product safety includes:
 Safety of test
 Safety of germs

Safety of life:
Safety of life includes:

 Safety of environment (lab, surroundings)

  Safety of personnel (Lab attendants, Lab assistants, supervisors, directors)
 Safety of surrounding populations (humans, animals, plants, faunna, flora)

Biosafety measurements in Microbiology Lab:

Following are the biosafety measurements in Microbiology lab:
i. Microbes causediseases so you should avoid direct contact with microbes.
ii. Chemicals are used to identify germs, these chemicals are toxic. You should avoid direct contact
with chemicals.
iii. Instruments of microbiology lab are costly. They should not be mishandled.
iv. Protection should be given to surrounding humans. You should not spread germs and chemicals.

Biosecurity:
Biosecurity means to provide security to personnel, building and infrastructure while working.
Infrastructure includes:

 Lock and key (guard,camera,thumb impression, retina scan)

 Equipment
 Chemicals
 Biological tests/Germs

Points:
 Biosafety is the protection of tests/germs in the lab.
 Biosecurity includes protection of tests/germs against notorious physical factors.
 Typhoid XDR:
 Disease spread in Karachi
 Disease spread through flies/ not properly cooked meat.
 Extreme drug resistance is produced by microbes causing the disease.

LECTURE NO.01:
Morphology of microorganisms:

 Bacteria
 Virus
 Fungus
 Algae

Bacteria:
Every bacterium has a unique shape.

Cocci (sing. Coccus):

Round shaped bacteria are called cocci.

Size:
Size ranges from 0.5 micrometer to 1 micrometer.

Example:

 Mycoplasma ( micrococcus)
 Staphylococcus (cocci)
 Division of cocci:
There are four main divisions of cocci:
i. staphylococci:
 Reproduce three dimensionally
 Grape like clusters
 For example, Staphylococcus
ii. streptococci:
 Reproduce in one dimension/single plane
 Form chains
 For example, Streptococcus
iii. tetrad:
 When cocci divide in two dimensions ( length and width ) they form tetrad.
 For example, Tetrad
iv. sarcina (p.sarcinae):
 When cocci divides in three dimensions at the same time they form a cube like structure
called sarcina.

Bacilli (sing. Bacillus ):

 Staff-like/Rod like bacteria

Size:

 Ranges from 3 micrometer to 10 micrometer in length.

 Ranges from 0.5 micrometer to 1 micrometer in width.

Division of bacillus:
There are three main divisions of bacillus:
i. streptobacillus:
 Long chains
 Divide by width
 For example, Bacillus
ii. Chinese letter formation:
 For example, Corynibacterium
iii. palisade:
 Stacks of coins
 Divide by length
 For example, Palisade
Sometimes bacteria have:
Filamentous / Branching shape:
 Like hairs

Size:

 More than 10 micrometer

 For example, Actinomycetes, Nocardia

Corkscrew:

 Spiral shaped
 For example, Treponema, Spirilla

Comma shaped/vibrio:
  For example, Vibrio fetus( humans )
 Campylobacter (animals)

Square shaped bacteria:

 For example, hollacrula

Star shaped bacteria:

 For example, Stella

Viruses:
According to morphology viruses are of five types:

i. Enveloped:
 Icosahedral:
 Envelope is circular or disorganized.
 Size is 100nm
 For example Herpes viridae
 Some have size more than 100 nm (e.g. Orthomyxoviridae)
ii. Naked:
 Icosahedral (without envelope)
 Size is 30nm to 100nm
iii. Complex:
They are of two types:
i. Pox viruses:
 Large brick like
 Size is 300nm to 1500nm
ii. Bacteriophage:
 Size is 50nm
iv. Filamentous viruses:
 They have length in mm
 They have width in nm
 For example , Filonidae
v. Mega viruses:
 Live in insects and protozoans
 Not important for humans and animals
 Size is in micrometers
 For example, Megavirales

Fungus:
Fungus is of two types:
1. Unicellular
2. Multicellular
 Unicellular:
 Unicellular fungus is called yeast.
 They are round in shape
 Their size is 5 micrometer to 10 micrometer
 Overall range of size is 5 micrometer to 40 micrometer
 Maximum size is 40 micrometer
 For example, Saccharomyces
 Multicellular:
 They have mycelium ,aerial spores and aerial hyphae
 Size is more than 10 micrometer
 For example, Aspergillus
  Some fungi are dimorphic/ bimorphic be cause they can exist as bot h unicellular and
multicellular at different temperatures.

Algae:
 Diverse in size
 Used as food source
 Photosynthetic
 They can cause ning but yeast cannot cause p oisoning
poiso to 40 micrometer
 Size is 5 micrometerve the size of 5 micrometer
 Blue Green algae ha eterinary importance up till now.
 There isno algae of v

E NO. 02:
LECTUR
Prokaryotes vs. Eukaryotes
Sr. No. Prokaryotes Eukaryotes
Term Before nucleus True nucleus
Primitive Modern

Groups Bacteria, Archeobacteria Algae, fungi, plants,

animals

Origin 3.5 billion years ago 2 billion years ago

Size 0.5 - 3 micrometer More than 5 micrometer
Cells Unicellular Unicellular/ Multicellular
Complexity Simple Complex

Nucleus location Free Center

 Free, attached to  Bounded in

cell membrane nucleus
 Circular  Linear
 One in bacteria,  Two or more than
DNA two in some two in Eukaryotes
 Replication,  Replication and
transcription, translation takes
translation takes place in cytoplasm,
place in cytoplasm transcription takes
at the same time place in nucleus
 Cell membrane but not at the same
aids in genomic time
process  No aid by cell
membrane in
genomic process

Nuclear membrane None Yes

Nucleolus Absent Present
Chromosomes Usually one More than one

Chromosome shape Circular Linear

 Operon Individually
Genes Multiple genes for a Single gene for single
single process process

DNA bases (G+C ratio) 28 - 73 40

DNA wrapping Non histonic proteins Histone proteins

Absent Present
 Inclusion bodies (Nucleus, E.R, Golgi
Membrane based are present. apparatus, lysosome,
organelles  Respiration takes mitochondria etc)
place at the cell
membrane
Ribosomes 70S (50S+30S) 80S (60S+40S)
Ribosomal region Cell membrane or free Endoplasmic reticulum
Mitochondria Absent Present

Chloroplast If present, chlorophyll is Present, chlorophyll within

scattered chloroplast
Microtubules/
cytoskeleton/ cell Simple or absent Present
movement
Cell membrane No sterols Sterols

Cell wall Peptidoglycan or muramic Starch/ Chitin

acid

Reproduction Simple/ binary fission Complex (2 gametes)

Basal metabolic rate High Low
Fimbrae/ flagella/ cillia Simple Complex

LECTURE NO.03:
Introduction to Microbiology
The word “Microbiology" is derived from Greek words:

MIKROS → small

BIOS → life
LOGIA → study

 It is the study of microorganisms either unicellular, multicellular or acellular.

 Microbiology includes disciplines of Virology, Mycology, Bacteriology etc.

Prokaryotic Vs. Eukaryotic Organisms:

 Prokaryotic microorganisms
are conventionally classified as
lacking organelles and includes:
→Eubacteria
→Archaebacteria
→Blue Green Algae
 Eukaryotic microorganisms exhibit cell organelles and include:

→Fungi

→Protists

→Algae

Additional Points:

 Only 1% of microorganisms are studied by culturing.

 The microorganisms which can be grown in culture media are studied by using
Molecular Biology. Their molecular study includes study of DNA, RNA & genes.
  Prions
caused a
disease
called
“Wasting
Dear/Zombi
e Dear disease in America.

How Microbiologists Study Microbes:

Microbiologists traditionally relied on culture, staining and microscopy. However, 1% of
Microbes present in the environment are culturable. That's why Microbiologists often
rely on extraction or detection of nucleic acids either DNA or RNA.
Viruses & Prions:
 Viruses are not always classified as organisms as they have been identified either
as very simple microorganisms or very complex molecules.
 Prions are never considered as microorganisms and they have been investigated
by the virologists as the clinical effects traced to them were originally presumed
due to chronic viral infections and virologists identified them as “infectious agents.
Following are some infectious
→Mad cow disease in cattle.
→Scrapie disease in sheep.
→Wasting deer / Zombie deer disease in deers (America).

BRANCHES OF MICROBIOLOGY:
Two broad branches of Microbiology are as follows:
1. Pure Microbiology
2. Applied Microbiology
Pure Microbiology:
It includes the study of Microbes only for the sake of
study. No benefits or loses are studied. Pure
Microbiology has further seven divisions:

1. Bacteriology:
Study of bacteria is called Bacteriology.

2. Mycology:
Study of fungi is called Mycology.
3. Phycology:
Study of algae is called Phycology.

4. Immunology:
Study of immune system is called immunology.
5. Virology:
Study of viruses is called virology.

6. Microbial Physiology:
Study of microbial cell functions is called Microbial Physiology. It includes the study of
microbial growth, metabolism & cell functions.

7. MicrobialCytology:
Study of microscopic & sub-microscopic details of microorganisms is called Microbial
Cytology.
 APPLIED MICROBIOLOGY:
Study of microorganisms to determine benefits as well as
harmful effects of microorganisms.
Applied Microbiology is further divided into four branches:
1.Veterinary Microbiology:
Study of microbes of Veterinary importance such as probiotics & pathogens which
are related to Veterinary medicine.

2.Pharmaceutical Microbiology:
Study of microorganisms which are related to the production of antibiotics,
enzymes, vitamins, biologics and other pharmaceutical products & those microbes
which cause pharmaceutical contamination & spoilage.
3. Microbial Biotechnology:
Manipulation of microorganisms at genetics & molecular level to generate useful
products is called Microbial Biotechnology e.g. production of insulin from E.coli.

4. Food Microbiology:
Study of microorganisms causing food spoilage and food
borne illness. Use of microbes to produce food is called food Microbiology.
  HISTORY OF MICROBIOLOGY:
Ancient History of Microbiology:
 Recent discovery of Mycobacterium DNA in the three thousand years old
Egyptian mummies reminds us that microorganisms have been around for a
much longer period of time.
 Infact, bacterium ancestors were the first living cells to appear on the earth.

Golden Age of Microbiology:

From 1857-1940 has been named Golden age of Microbiology. During this period, rapid
advancement spread hold mainly by Pasture & Robert Koch which led to the
establishment of
Microbiology as a science. During this era, salient achievements include;

 Discovery of immunity
 Discovery of disease causing agents
Koch's Postulates:
 Robert Koch a Germen physician discovered the cause of Anthrax (Bacillus
anthracis) in 1870s.
 He devised a theory relating micro-organisms to every specific disease. The
POSTULATES of that theory are as under.
 Same pathogen must be present in every case of the disease.
 Pathogen must be isolated from the diseased host & grown in a pure culture.
 Pathogen from the pure culture must cause the disease when it is inoculated into
susceptible lab. animal.
 The same pathogen must be isolated from the inoculated animal and it must be
same as original organism.

Vaccination:
Edward Jenner, a young British physician used scrapping of cowpox blisters to
vaccinate against smallpox.
 Edward Jenner is called the father of Vaccinology.
Fermentation:
 Microorganisms like yeast convert sugar into alcohol in the absence of air. This
process is called Fermentation.
 It is used to make wine & beer.
 In the presence of air bacteria converts alcohol into acetic acid or vinegar.

Pasteurization:
 It is the heat treatment of beverage & milk at 72°C for 30 minutes to kill
microorganisms (which cause spoilage of liquid in food) without compromising
on its quality.
 Now a day, it is only used for milk products.

Germ Theory of Disease:

 Microorganisms are the cause of diseases. In 1860's Joseph Lister used phenol,
carbolic acid as a disinfectant & antiseptic solution.
 This practice reduced the incidence of infection & death. Other surgeons readily
adopted it.
 Lister's technique was the earliest medical attempt to control infection caused by
microorganisms.
LECTURE NO.04:
SIZE OF BACTERIA:
Many sizes and most bacteria range from:

 0.2 to 2.0 μm in diameter

 2 to 8 μm in length
 Overall average size 1-10 μm .

SHAPE OF BACTERIA:
 Spherical / coccus (plural: cocci; means rounded)
 Rod Shaped / bacillus (plural: bacilli)
 Spiral
Cocci:
Cocci are usually round but can be oval or elongated. When they divide to reproduce,
they remain attached to each other.

CLASSIFICATION ACCORDING TO PLANE OFDIVISION:

Dipplococci:
Those cocci that remain in pairs after dividing are called diplococci.

Streptococci:
Those cocci that divide and remain attached in chainlike patterns are called streptococci.
Tetrad:
Those cocci that divide into two planes and remain in groups of four are known as
tetrads.

Sarcinae:
Those cocci that divide in three planes and remain attached in cube like groups of eight
is called sarcina.

Staphylococci:
Those cocci that divide in multiple planes and form grapelike clusters or broad sheets
are called staphylococci.

BACILLI:
Bacilli are the rod like bacteria e.g. Bacillus anthracis

Single bacillus:
Most bacilli appear as single rods, called single bacilli.
Diplobacilli:
Those bacilli which appear in pairs after division are called Diplobacilli.

Streptobacilli:
Those bacilli that are found in chains in single plane are called Streptobacilli.

MONOMORPHIC VS PLEOMORPHIC:
 Bacteria that maintain a single shape are called monomorphic or the bacteria
having same shape.
 Some bacteria can have many shapes known as pleomorphic. e.g.:
Corynebacterium pyogenese.
 Some bacteria occur in the shape of a star e.g. Stella.
 Some bacteria occur in the shape of rectangle e.g. Haloarcula.
 Some bacteria also appear triangular in shape.

FUNCTIONAL ANATOMY OF BACTERIA:

Cell Wall:
Acts as an antigen, provide protection and rigidity

Cell membrane:
Serve as a barrier through which materials enter and exit the cell

Capsule:
Acts as an antigen, has feeding importance, sticking features and cause disease

Mesosomes:
Has role in metabolism
FUNCTIONAL ANATOMY OF BACTERIA:
Fimbrae:
Helps in motility, jerky movement, has sticking feature and acts as an antigen

Pilus:
Helps in reproduction (conjugation)

Ribosomes:
Protein formation

Nucleoid:
Transcription and translation

FUNCTIONAL ANATOMY OF BACTERIA:

Chromosomes:
Hereditary material

Droplets:
Helpful in storage

Flagella:
Act as an antigen and helps in motility

Plasmid:
Have special features of resistance and infection.