Scribd
Upload
13 views31 pages

Prac. Microbiology

Uploaded by

Zeena Muhamed
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
13 views31 pages

Prac. Microbiology

Uploaded by

Zeena Muhamed
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
You are on page 1/ 31

microbiology lab 4

Culture Media
Culture Media

 • To study about the pathogens and commensal organisms of the


human body, they need to be cultivated on artificial culture media.

 • Culture medium is material which provides physical and chemical


requirement for microbial growth.

 • Physical requirement include heat, ph, osmotic pressure.

 • Chemical requirement include carbon, nitrogen, phosphorus, sulfur,


minerals, vitamins and water.
Classification of Culture media

 Bacterial culture media can be classified in different ways; based on


consistency, based on nutritional component and based on its functional
use.
A) Classification based on consistency
• Culture media are liquid, semi-solid or solid and semisolid.
1. Liquid media (Broth) :-
 • no agar
 • Liquid media tend to be used when a large number of bacteria have to be grown.
 • e.g. Nutrient broth
2. Solid media :- ( 2 % agar)
 • liquid medium with addition of solidifying agents such as agar-agar,
egg
yolk or serum.
 • it can be used for culture isolation, colonial appearance and tests for
extracellular enzymes.
• E.g. : Nutrient agar, Blood agar.
3. Semi-solid media:- (0.5 % agar)
• useful in demonstrating bacterial motility and separating motile from
non-motile strains
 • Agar-agar is a complex polysaccharide extracted from marine algae
agar, is not digested by most microorganisms, melt at 100 c and solidify at
45 c.
B) Classification based on nutritional component :-

• Media can be classified as simple, complex and


synthetic .
 1. Simple media :-
• such as peptone water, nutrient agar can support most
non-fastidious bacteria.
 2. Complex media :-
 • such as blood agar have ingredients in which the
exact components are difficult to estimate.
 3. Synthetic media :-
• specially prepared media for research purposes where
the composition of every component is well known
C) Classification based on functional use or application :-
1. Basal media :-
• Basically is simple media that supports most non-
fastidious bacteria.
• nutrient broth and nutrient agar are considered as basal
media.
2. Enriched media
• Substances like blood, serum, egg are added to the
basal medium.
• used to grow nutritionally exacting (fastidious) bacteria.
• E.g. : Blood agar, Chocolate agar
 3. Enrichment media :-
• liquid media which encourage the growth of certain kinds of
bacteria in mixed population.
 • Stimulate growth of desired bacterium
 • Inhibit growth of unwanted bacterium
• E.g. :-
Selenite F Broth – for the isolation of Salmonella, Shigella
Tetrathionate Broth – inhibit coliforms
Alkaline Peptone Water – for Vibrio cholera
4. Selective media :-
• These are designed to inhibit unwanted commensal or contaminating
bacteria
• It does not enhance the growth of any bacteria.
• addition of antibiotics, dyes, chemicals, alteration of pH or a
combination of these.
 • E.g.:
Desoxycholate citrate medium for dysentery bacilli
Mac Conkey’s medium for gram negative bacteria
TCBS – for V. cholerae
Lowenstein–Jensen medium – M. tuberculosis
TCBS
Mac Conkey’s medium
5. Differential media or indicator media :-
• In this media bacteria can be recognized on the basis of their colony
colour.
• incorporation of dyes, metabolic substrates etc.
 • Eg:-
Wilson-Blair medium – S. typhi forms black colonies
MacConkey’s medium - Distinguish between lactose fermenters & non
lactose fermenters.
Blood agar
6. Transporting media :-
• Such media prevent drying of specimen, maintain the pathogen to
commensal ratio and inhibit overgrowth of unwanted bacteria.
• Eg:
• Stuart’s medium – non nutrient soft agar gel containing a reducing
agent & charcoal (neutralize inhibitory factors)
• Buffered glycerol saline – enteric bacilli
7. Anaerobic media :-
• Anaerobic bacteria need low oxygen content, reduced
oxidation –reduction potential and extra nutrients.
• Have specific substances that absorb oxygen
• Eg:
Robertson’s cooked meat medium ( for Clostridium spp.)
Thioglycolate medium.
Methods of Culturing

1. Streak culture.


2. Lawn culture.
3. Stroke culture.
4. Stab culture.
5. Pour plate culture.
6. Liquid culture
1. Streak culture :-
• Routinely used method to isolate bacteria.
• One loop-full of culture is made as a primary inoculum
and is then distributed
thinly over the plate by streaking it with the loop in a series
of parallel lines in different segments of the plate.
2. Lawn culture :-
• Useful for bacteriophage typing and antibiotic sensitivity testing.
• Prepared by flooding the surface of the plate with a liquid culture or
suspension of the bacterium.
• Alternatively the surface of the plate may be inoculated by applying a swab
soaked in the bacterial culture or suspension.
 3.Stroke culture :-
• Stroke culture is made in tubes containing agar slope .
• It provides a pure growth of bacterium for slide agglutination and
other diagnostic tests.
4.Stab culture :-
• Stab cultures in solid media are inoculated by plunging the straight
wire into the center of the medium and with drawing it in the same
line to avoid splitting the medium.
5. Liquid culture :-
• Liquid cultures are inoculated by touching with a charged loop or
by adding the inoculum with pipettes or syringes.
• Uses
Blood culture
Continuous culture methods
 6 pour plate culture

You might also like

pFad - Phonifier reborn

Pfad - The Proxy pFad of © 2024 Garber Painting. All rights reserved.

Note: This service is not intended for secure transactions such as banking, social media, email, or purchasing. Use at your own risk. We assume no liability whatsoever for broken pages.


Alternative Proxies:

Alternative Proxy

pFad Proxy

pFad v3 Proxy

pFad v4 Proxy