The document outlines the nutritional requirements, preparation, classification, and physical parameters for culture media used in cultivating microorganisms. It details various types of media, including basic, selective, differential, enriched, and transport media, highlighting their components and specific uses in microbiology. Key components for growth are also addressed, including chemical and physical requirements such as nutrients and environmental conditions.

1. Nutritional Requirements, Raw
Materials Used For Culture Media
And Physical
Parameters For Growth
Prepared By-
D.Mahendra,M.Pharm,(Ph.D),.
Dept of PA & QA

2. Subjects included;
• Cultivation
• Medium
• Inoculation of media
• The Requirements for Growth
• Preparation
• Classification
• Eplanation of types of media
functionally.

3. Cultivation;
The propagation of living organisms, applied
especially to the growth of microorganisms or other
cells in artificial media.
Medium
Any preparation that contains nutrients essential for
microorganisms growth.
Culture medium:
A nutrient substance that is used to grow
micro-organisms .

4. Inoculation of media
Introduction of infected material to the medium ,for
cultivation of organisms.
Agar
Is Complex polysaccharide, Used as solidifying
agent for culture media in Petri plates, slants, and
deeps. Generally not metabolized by microbes
Liquefies at 100°C
Solidifies at ~40°C

5. The Requirements for Growth
1.CHEMICAL
REQUIREMENTS
(NUTRITIONAL FACTORS)
• Carbon
• Nitrogen, sulfur, and phosphorous
• Trace elements
• Organic growth factor
• Vitamins (e.g. folic acid, vitamin B-12, vitamin K)
• Oxygen

6. 2.PHYSICAL REQUIREMENTS
• Temperature
• PH
• Hydrostatic Pressure
• Osmotic pressure

7. Preparation
• Wash hands & wear gloves
• Sterilize all equipments
• Pour D/W in glassware (required amount)
• Add powder ingredients (required amount)
• Heat to dissolve completely
• Autoclave
• Dispense the medium into tubes ,bottles
&plates
• Store at required temperature
• Sterilizing Culture Media

8. Classification
1. Consistency
2. Nutritional
3. Functional

9. 1.Consistency
a ) Solid e.g nutrient agar
b ) Semi-solid e.g peptone water.
c ) Liquid medium e.g nutrient broth.

10. 2.Nutritional
a ) Simple e.g peptone water
b ) Complex e.g blood agar
c ) Synthetic e.g Davis /mingioli medium

11. 3.Functional
a) Basic media
b) Selective media
c ) Differentialmedia
d ) Enriched media
e ) Enrichment media
f ) Transport media

13. Basic media
• This is an undefined medium because the
amino acid source contains a variety of
compounds with the exact composition
being unknown.
• Nutrient media contain all the elements
that most bacteria need for growth and are
non-selective, so they are used for the
general cultivation and maintenance of
bacteria kept in laboratory culture
collections.

14. Basic media
• This media contains;
1.A carbon source such as glucose for
bacterial growth
2.Water
3.Various salts needed for bacterial growth

15. 1. nutrient broth which is liquid media
There are two types:

16. 2. nutrient agar which is solid media

17. Nutritional Requirements for Agar
Media (Composition)

18. Selective media
• culture medium that allows the growth of
certain types of organisms, while
inhibiting the growth of other organisms
• Any agar media can be made
selective by addition of certain
inhibitory agents that
don’t affect the pathogen.
• Various approaches to make a
medium selective include addition
ofantibiotics, dyes, chemicals,
alteration of pH or a combination of
these medium

19. Differential media
• distinguishs one microorganism type from
another based on a difference in colony
appearance includes (color, shape, or
growth pattern) on the media.
• uses the biochemical characteristics of a
microorganism growing in the presence of
specific nutrients or indicators (such as
neutral red, phenol red , or methylene blue)
added to the medium to visibly indicate the
defining characteristics of a microorganism

20. Differential media
• is used for the detection of microorganisms
and by molecular biologists to detect
recombinant strains of bacteria
• Examples;
Mannitol salt agar
Eosin methylene blue
MacConkey agar

21. • Mannitol salt agar
which is differential for mannitol
fermentation
Like Staphylococcus aureus change
the color of agar from pink to yellow
• Eosin methylene blue
which is differential for lactose
and sucrose fermentation

22. • MacConkey agar
which is differential for lactose fermentation Ex: E.coli
From non lactose fermentersEx: salmonella,shigella

23. Enriched media
• Enriched with Blood,serum,extrapeptones,egg&
vitamins
• Used for:
cultivation of all fastidious organisms, (haemophilus
influenza&Streptococcus spp.)
• Examples:
• Blood agar &chocolate agar

24. 1.agar powders
t
2.Weighing out
he ingredients
3.Measuring
the desired
volume of
water for the
medium
5.Autoclaving
6.Adding
blood to
blood agar
base
medium
4.7A.Mddixininggthe
cothnetebnltosotodbottle
with
molten
medium
at 45 c 8.Pouring
plates
9.Laying out
plates to set
Preparation of
bloodmedia

25. Enrichment media
• A liquid culture medium promotes the
growth of a particular organism and
prevent the growth of normal
competitors.
• Prevent non pathogenic bacteria from
overgrowing the pathogenic bacteria
• Example: selenite-Fbroth
alkaline-peptone water

26. Transport media
• Preserve viability and ratio of
microbes during its transportation
over 24_48 hours.
• Example:
CARY-BLAIR MEDIUM
STUART TRANSPORT MEDIUM

27. CARY-BLAIR MEDIUM
• Intended use;
• for the collection and transport of fecal and rectal
samples, maintaining viability of Salmonella and
Shigella.
• Survival depends upon:
1.bacterial type &concentration , transport medium
formulation, transport temperature and duration.
• the color is white opalescent

28. • Intended use:
is a semisolid medium used in the transport and
preservation of specimens for the cultivation of
diverse organisms such as gonococci,
streptococci, Enterobacteria
The color : is blue insurface.
STUART TRANSPORT MEDIUM