Culture media are used to grow microorganisms outside their natural environment in the laboratory. They provide nutrients to support microbial growth. Common media include nutrient broths and agar plates. Specialized media are sometimes needed to cultivate specific microbes. Media can be classified based on physical state (liquid, semisolid, solid), presence of oxygen, nutritional components (simple, complex, synthetic), and purpose (enrichment, selective, differential). Selective media inhibit all bacteria except the desired type, while differential media help distinguish between microbial species.
he culture media are classified in many different ways: Based on the physical state Liquid media Solid media Semisolid media Based on the presence or absence of oxygen Anaerobic media Aerobic media Based on nutritional factors Simple media Synthetic media Complex
he culture media are classified in many different ways: Based on the physical state Liquid media Solid media Semisolid media Based on the presence or absence of oxygen Anaerobic media Aerobic media Based on nutritional factors Simple media Synthetic media Complex
Culture media are the basic requirement to grow the microbes in the laboratory for various purpose like isolation, identification and research purposes.
These slides explain how media preparation in microbiology lab
for bacterial culture, history of culture media, types of culture media based on concentration and ingredients and practical section.
This ppt is on culture media used in microbiology study. Topics covered~ What is culture media?, Major contribution of scientist, Types of culture media~ physical nature, chemical composition, Basic requirements, Functional types, supportive media, media composition, enriched media, selective media, MacConkey agar composition, Differential media, selective & indicator media.
culture media
CULTURE – Is term given to microorganisms that are cultivated in the lab for the purpose of studying them.
MEDIUM – Is the term given to the combination of ingredients that will support the growth & cultivation of microorganisms outside their natural habitats.
Necessary Requirements for Growth of Bacteria
Distilled Water
Nitrogen containing compounds
Peptone- Golden granular powder
Complex mixture of partially digested protiens by proteolytic
enzymes pepsin, trysin or papain
Peptones, Proteoses, polypeptides, aminoacids, inorganic salts like phosphates
potassium & magnesium
Accessory growth factors like nicotinic acid & riboflavin
Energy sources
Suitable Ph- 7.2 – 7.4
Solidifying agents:
Gelatin– Protien
Agar— Chief component is Long chain Polysaccharide
Melts at 95°c & solidify only when cooled to about 42°c
1- 2% yields a suitable gel eg. Non-nutritive agar
According to Physical State:
Liquid – Peptone Water, Nutrient Broth
Semisolid – Nutrient Agar Stabs
Solid – Blood Agar
According to Oxygen requirement:
Aerobic Medium
Anaerobic Media
The process of growing microorganisms in culture by taking bacteria from the infection site (in vivo or environment) and grow them in artificial environment in the laboratory (in vitro).
Bacteria may require adequate nutrition, optimum pH, temperature and oxygen for growth and multiplication.
Suitable artificial media containing sources of carbon, nitrogen, hydrogen, oxygen, phosphorous and other elements such as sodium, potassium, magnesium, iron and growth factor (Vitamins) in very small amounts have been used for cultivation of microorganism.
When microorganisms are cultivated in the laboratory, a growth environment called a medium is used. The medium may be purely chemical (a chemically defined medium), or it may contain organic materials, or it may consist of living organisms such as fertilized eggs.
Microorganisms growing in or on such a medium form a culture.
A culture is considered a pure culture if only one type of organism is present and a mixed culture if populations of different organisms are present.
When first used, the culture medium should be sterile, meaning that no form of life is present before inoculation with the microorganism.
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Culture media are the basic requirement to grow the microbes in the laboratory for various purpose like isolation, identification and research purposes.
These slides explain how media preparation in microbiology lab
for bacterial culture, history of culture media, types of culture media based on concentration and ingredients and practical section.
This ppt is on culture media used in microbiology study. Topics covered~ What is culture media?, Major contribution of scientist, Types of culture media~ physical nature, chemical composition, Basic requirements, Functional types, supportive media, media composition, enriched media, selective media, MacConkey agar composition, Differential media, selective & indicator media.
culture media
CULTURE – Is term given to microorganisms that are cultivated in the lab for the purpose of studying them.
MEDIUM – Is the term given to the combination of ingredients that will support the growth & cultivation of microorganisms outside their natural habitats.
Necessary Requirements for Growth of Bacteria
Distilled Water
Nitrogen containing compounds
Peptone- Golden granular powder
Complex mixture of partially digested protiens by proteolytic
enzymes pepsin, trysin or papain
Peptones, Proteoses, polypeptides, aminoacids, inorganic salts like phosphates
potassium & magnesium
Accessory growth factors like nicotinic acid & riboflavin
Energy sources
Suitable Ph- 7.2 – 7.4
Solidifying agents:
Gelatin– Protien
Agar— Chief component is Long chain Polysaccharide
Melts at 95°c & solidify only when cooled to about 42°c
1- 2% yields a suitable gel eg. Non-nutritive agar
According to Physical State:
Liquid – Peptone Water, Nutrient Broth
Semisolid – Nutrient Agar Stabs
Solid – Blood Agar
According to Oxygen requirement:
Aerobic Medium
Anaerobic Media
The process of growing microorganisms in culture by taking bacteria from the infection site (in vivo or environment) and grow them in artificial environment in the laboratory (in vitro).
Bacteria may require adequate nutrition, optimum pH, temperature and oxygen for growth and multiplication.
Suitable artificial media containing sources of carbon, nitrogen, hydrogen, oxygen, phosphorous and other elements such as sodium, potassium, magnesium, iron and growth factor (Vitamins) in very small amounts have been used for cultivation of microorganism.
When microorganisms are cultivated in the laboratory, a growth environment called a medium is used. The medium may be purely chemical (a chemically defined medium), or it may contain organic materials, or it may consist of living organisms such as fertilized eggs.
Microorganisms growing in or on such a medium form a culture.
A culture is considered a pure culture if only one type of organism is present and a mixed culture if populations of different organisms are present.
When first used, the culture medium should be sterile, meaning that no form of life is present before inoculation with the microorganism.
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2. • CULTURE :
Is the term given to microorganisms that
are cultivated in the lab for the purpose of
identifying and studying them.
2
CULTURE AND THE MEDIUM
3. • MEDIUM:
Is the term given to the combination of
ingredients that will support the growth and
cultivation of microorganisms by providing
all the essential nutrients required for the
growth (i.e. multiplication) in order to
cultivate these microorganisms in large
number to study them.
3
4. • Microbiological culture: which are
used for growing microorganisms,
such as bacteria or yeast.
• The most common growth media for
microorganisms are nutrient broths and agar
plates
• Specialized media are sometimes required for
microorganism and cell culture growth.
4
5. CULTURE MEDIA
• Used to grow bacteria
• Can be used to-
Enrich the numbers of bacteria.
Select for certain bacteria and suppress others.
Differentiate among different kinds of bacteria.
5
6. NEED FOR CULTURE MEDIA
• It is usually essential to obtain a culture by
growing the organism in an artificial medium.
• If more than one species or type of organism are
present each requires to be carefully separated or
isolated in pure culture .
10/27/2023 6
7. BASIC REQUIREMENTS OF CULTURE MEDIA
• NUTRIENTS :
Energy source
Carbon source
Nitrogen source
• MINERAL SALTS :
Sulphates, phosphates, chlorides and carbonates
of K, Mg and Ca
A suitable pH- 7.2- 7.4
7
8. CLASSIFICATION
BASED ON
PHYSICAL
STATE
BASED ON
PRESENCE OF
MOLECULAR
OXYGEN AND
REDUCING
SUBSTANCES
BASED ON
NUTRITIONAL
FACTORS
LIQUID MEDIA AEROBIC MEDIA SIMPLE MEDIA
SEMISOLID MEDIA ANAEROBIC MEDIA COMPLEX MEDIA
SOLID MEDIA SYNTHETIC MEDIA
SPECIAL MEDIA
8
9. SPECIAL MEDIA
A. ENRICHED MEDIA
B. ENRICHMENT MEDIA
C. SELECTIVE MEDIA
D. DIFFERENTIAL MEDIA
E. INDICATOR MEDIA
F. TRANSPORT MEDIA
G. SUGAR MEDIA
9
10. SIMPLE MEDIA
Simple or basal media are culture media which contain the minimum
adequate nutrition for non fastidious organisms
Example:- Nutrient broth/agar
Peptone water
Composition:-
Lab-Lemco -10gm
peptone-10gm
NaCl- 5gm
Distilled water-1000ml
- When 2-3% agar is added ,then we have it as nutrient agar.
- For semisolid media – agar concentration is 0.2-0.4%
Uses:-
1. This is basis of most of the media used in the study at common
pathogenic bacteria.
2. It is used for subcultures of certain organisms.
12. PEPTONE WATER
• TYPE : Basic liquid media
• APPEARANCE : clear, colorless, watery, usually
in test tube
• Composition :
12
PEPTONE 10 g
SODIUM CHLORIDE, NaCl 5 g
WATER 1 litre
13. USES OF PEPTONE WATER
The media is used chiefly as the basis for
carbohydrate fermentation media.
Nutrient broths may contain a small amount
of sugar derived from meat and it is essential
that the basal medium to which various
carbohydrates are added for fermentation
tests should be free from natural sugars.
10/27/2023 13
14. It is also used to test the formation of indole.
Culture of organisms for demonstration of motility
10/27/2023 14
15. COMPLEX MEDIA
Complex media have added ingredients for bringing out certain properties
for bringing out certain properties or providing special nutrients required
for growth of the bacterium in question.
SYNTHETIC MEDIA
These are prepared from pure chemicals and the exact compositions of
medium is very well known.
Example :- Dubo’s medium
SEMIDEFINED MEDIA
In these media the exact chemical composition of the constituents is not
known because substances like meat and peptone are used.
Most of the culture media used for routine diagnostic work are
semidefined culture media.
16. SPECIAL MEDIUM
• ENRICHED MEDIA
When basal medium is added with some nutrients such as blood,
serum or egg is called enriched media.
They are used to grow bacteria which are more exacting in their
nutritional needs.
Examples:-
Dorset’s Egg Medium.
It is a creamy coloured opaque
slope kept in screw copped bottle
Selective medium for isolation
of Mycobacterium tuberculosis.
Composition: Hen’s egg, Nutrient broth
17. BLOOD AGAR
• TYPE : Enriched media.
• APPEARANCE : Red color.
• COMPOSITION :
Sterile Nutrient agar + Defibrinated sheep blood
USES :
Routine culture
Widely used in medical bacteriology
It is also an indicator medium showing the haemolytic
properties of bacteria such as Streptococcus pyogenes.
17
18. CHOCOLATEAGAR
Also called Heated blood agar.
• TYPE : Enriched media.
• APPEARANCE : Chocolate brown color.
PROCEDURE
Melt the desired amount of nutrient agar.
Cool it in a water – bath at 75º C .
Add 10 ml of sterile blood .
Allow the medium to remain at 75º C.
18
19. Mixing the blood and agar by gentle agitation from
time to time until the blood become chocolate
brown in color, within about 10 min.
Then pour in plates.
USES
CULTURE OF Neisseria
CULTURE OF Haemophilus influenzae
CULTURE OF Pneumococcus
19
20. ENRICHMENT MEDIA
In this media, it has a stimulating effect on the bacteria to be grown or inhibits
its competitors.
This result in an absolute increase in the number of wanted bacteria related to
other bacteria.
Example:- Selenite F broth
It is enrichment medium for culture of Salmonella typhi and paratyphi bacilli
from stool sample
Principle:- at neutral pH solution acid salinity has high toxicity to coli form
group of bacteria and not to most of the salmonella groups.
22. SELECTIVE MEDIA
It is a medium in which certain substances are present which inhibit all other
bacteria except the desired bacteria.
It encourages the growth of particular species from a mixed inoculum.
Example:- TCBS
-It is light green translucent medium kept in petridish
-It is selective medium for Vibrio cholera
-Principle:-
Bile salt inhibit the growth of normal
commensals (unwanted bacteria).
Vibrio chloerae produce acid by fermentation
of sucrose which acts on bromothymol blue
(indicator) producing yellow colonies.
24. MAC CONKEY AGAR
• MacConkey agar is a culture medium
designed to grow Gram-negative bacteria. It is
a useful medium for the cultivation of
enterobacteriacea.
24
25. 25
MacConkey agar
showing both
lactose and non-
lactose fermenting
colonies.
Lactose fermenting
colonies are pink
whereas non-
lactose fermenting
ones are colourless
or appear same as
the medium.
26. ANAEROBIC MEDIUM
These media are used to grow anaerobic organisms.
Examples:-
Thioglycollate broth
Robertsons Cooked Meat Medium
28. • Mueller-Hinton agar is an microbiological
growth medium that is commonly used for
antibiotic susceptibility testing.
• Originally formulated for isolation of Neisseria
species.
• It is also used to isolate and maintain
Neisseria and Moraxella species.
28
BASED ON PHYSICAL STATE
Solid medium
Liquid medium
Semisolid medium
BASED ON PRESENCE OF MOLECULAR OXYGEN & REDUCING SUBSTANCE
Aerobic medium
Anaerobic medium
BASED ON NUTRITIONAL FACTORS
Simple medium
Complex medium
Synthetic or defined medium
Semidefined medium
Special medium
Enriched medium
Enrichment medium
Selective medium
Differential medium
Indicator medium
Transport medium
Sugar medium
pH – 7.4 to 7.5
Sterilization : sterilized by autoclaving at 121 degree C for 15 minutes
It is an enzymatic hydrolysate of animal tissues used as culture media ingredient in variety of media
Also useful for commercial production of enzymes, vaccines, antibiotics and other products.
Unsaturated fatty acid present in meat utilise oxygen for autoxidation this reaction is catalysed by haematin in the meat
Glutathione and cysteine present in meat also utilize oxygen
Sulphydryl compounds (present in cysteine) also contribute for a reduced oxidation – reduction potential
Procedure :- Before inoculation the medium is boiled in water bath at 80 degrees for 30 minutes to make oxygen free. For strict anaerobiosis the surface of CMB medium may be converted with a layer of sterile liquid paraffin
Interpretation:-
Sacchride anaerobes (Clostridium perfinges) turn the colour of meat pieces red while if become black in cases of proteolytic anaerobes (Clostridium tetani)