Escolar Documentos
Profissional Documentos
Cultura Documentos
Medical Faculty
Department of Microbiology, Virology and Immunology
MICROBIAL WORLD
Non Cellular Microbes
cellular
microbes
Prions Domain Domian MICROBIAL WORLD
Bacteria Archaea Domain Eukarya
Viriods
PROKARYOTES EUKARYOTES
Viruses
Bacteria Archaebacte Protozoa Fungi
ria
Microbes are too small to be seen with our naked
eye.
Bacilli
Cocci
Spiral
Filamentous
BAC
K
Bacilli
Cocci
Spiral
Filamentous
BAC
K
Bacilli
Cocci
Spiral
Filamentous
BAC
K
Bacilli
Cocci
Spiral
Filamentous
BAC
K
(Bacteriological Examinatio
1- obligate anaerobe
2- facultative anaerobe Aerobic
Anaerobic
3- microaerophile Culture
culture
4- strict aerobe
Microaerophili
c culture
1 2
3 4
Aerobic Culture
1 2
3 4
Anaerobic Culture
1 2
3 4
Microaerophilic culture
** Essential Media
It should contain essential food elements
It should have satisfactory moisture content
It should have correct pH (most bacteria are
neutrophiles. They grow best at pH between 6-8.
The consistency of the medium should be favourable
for cultivation.
It should be sterile.
It should be isotonic.
It should be transparent.
It should be oxygen-free if the cultivation of strict
anaerobes is desired.
Solid
Liquid
What we need for Inoculation??
Inoculating loops
START!!!
Petri-dishes with solid medium
Sterilize the inoculating loop Aseptic of instruments:
or needle by heating it in the
flame. Hold the instrument
like a pencil and at an angle
so that the wire is pointing
downward. Pass the entire
wire portion through the
flame starting at the base,
where it is attached to the
holder, and continue all the
way through the loop or end
of the needle. Heat the wire
until it is glowing red then
allow it to cool before
picking up any
microorganisms. The
instrument is now
considered to be sterile. Do
Streaking Technique:
2 2
1st quadrant
can be 3
1
inoculated with
a loop or swab.
Each of the
remaining three
quadrants must
be streaked 4
with a sterile
instrument
such as flamed
Pour Plate technique :
1.Dilute specimen to yield
approximately 30 to 300 CFU
per aliquot to be plated
2.Diluted sample is
added directly to the
empty plate
(Inoculation)
size
type of
margin
colony
elevation
colony
texture
Light
transmission
colony
pigmentation
2. Isolation
To obtain pure culture
Pure culture
colonies will be
If different types of obtained after
colonies can be seen on incubation.
the plate
EMB
Agar
Each of these plates
contain Gram negative
organisms that can
ferment lactose. The
left plate is
contaminated. Notice
the white colonies
interspersed with the
pink colonies. When
organisms ferment the
lactose in EMB they
produce acid which
turns the colonies pink,
purple or green,
depending on the pH.
Sheeps Blood Agar Plate
This media is used to test if an
organism has hemolysins
(enzymes that lyse red blood
cells). If an organism lyses red
blood cells in the media then
clearing zones are produced
around the culture. An organism
can be classified as alpha, beta or
gamma hemolytic. The culture
on the top portion of this plate is
showing alpha hemolysis (partial
clearing of red blood cells). The
culture on the bottom of the
plate is showing gamma
Triple Sugar Iron Slant (TSI
Slant)
TSI slants are used to test if an
organism ferments glucose and
sucrose and/or lactose and if the
can reduce sulfate. The tube on
the far left is uninoculated. The
second tube shows a yellow butt
and a yellow slant. This means
the organism fermented glucose
and lactose and/or sucrose. The
third tube has a yellow slant and
a yellow butt. This means that
the organism fermented glucose
and lactose and/or sucrose and
reduced sulfate. The fourth tube
from the left shows a yellow butt
and a red slant. This means that
the organism only fermented
Motility Test Media
3.
Identificatio
Identification of the pure culture of
n microbe on the basis of its cultural,
morphological and tinctorial
properties, biochemical and antigenic
characteristics
Cultural Examination (Bacteriological
method) allows microbiologist to
perform epidemiological typing of
isolated microbe and to choose the
drugs for antimicrobial therapy
Techniques for Epidemiology typing
1.Biotyping (biovars)
2.Serotyping (serovars)
3.Bacteriophage (phagovars)
4.Toxigenicity
5.Bacteriocin
6.Nucleic acid restriction analysis and other
genetic tests
7.Antibiograms
- Disk Diffusion Method
- Minimum Inhibitory Concentration (MIC)
Phagotyping Antibiogram
When pathogen cannot be isolated on the
culture media in laboratory, animated models
of culture media is used.
Incubation of
chicken
embryo
Negative
No reactions.
No contact with the
pathogens