CULTIVATION OF BACTERIA, NUTRITIONAL TYPES AND NUTRITIONAL CLASSIFICATION OF BACTERIA

1. CULTIVATION OF
BACTERIA, NUTRITIONAL
REQUIREMENTS AND
TYPES OF BACTERIA
NIMISHA.K
MSc FST

2. CULTIVATION OF
BACTERIA

3. The process of growing microbes in culture by;
 Taking bacteria from an infection site by specimen
Collection- in vivo
 Growing bacteria in the artificial environment of the
Laboratory- in vitro

4. PURPOSE OF THE CULTIVATION
• Isolate bacteria in pure culture
• Demonstrate their properties
• Determine sensitivity to antibiotics
• Maintain stock culture
• Obtain sufficient growth for preparation of
antigens and for other tests

5. TYPES OF
CULTURE MEDIA
A culture medium
is a mixture of
chemicals that can
support the growth
of micro
organisms, it
should contain a
source of carbon
and energy

6. 1. LIQUID MEDIA, SOLID MEDIA AND
SEMI SOLID MEDIA
• The liquid media contain peptone and NaCl as
the main ingredients
• The original media used by Louis Pasteur were
liquids such as urine or meat broth
• Eg:- Nutrient broth
• Solid media can be prepared by adding agar
into liquid media, agar (agar-agar) is act as
solidifying agent
• Eg:- Nutrient agar

7. Semi solid media
•Such media are fairly soft and are useful in demonstrating
bacterial motility
•A smaller percentage of agar is present, the agar is
sufficiently soft that motile bacteria can swim through the
media
Eg:- Semi solid nutrient agar

8. 2.SIMPLE, COMPLEX, DEFINED, SEMI
DEFINED, SPECIAL MEDIAS
Simple media (Basal media)
It consists of peptone, meat extract, sodium chloride and
water
It is the simplest and most common medium in routine
diagnostic laboratories
Eg:-Nutrient broth
Complex media
These have added ingredients for special purposes or for
bringing out certain characteristics or for providing
special nutrients required for the growth of bacterium
Eg:-Yeast extract agar

9. Synthetic or defined media
These media is prepared from pure chemical
substances and the exact composition of the
medium is fully documented
Eg:-Minimal media
Special media
• Enriched media:- In this media, substances such as
blood, serum or egg are added to a basal medium
Eg:-Blood agar

10. • Enrichment media:- Is a medium with specific and
known qualities that favors the growth of particular
microbes
Eg:- Selenite F Broth
• Selective media:-In this a inhibiting substance is
added, it enables a greater number of the required
bacterium to form colonies than other bacteria
Eg:- Thayer martin medium
• Indicator media:-These media contain an indicator
that changes color when a bacterium grows in them
Eg:-McLeod’s medium

11. • Differential media:-A medium that has substances
incorporated in it, enabling it to bring out differing
characteristic of bacteria and thus helping to distinguish
between them.
Eg:-MacConkey media
• Sugar media:-It consist of 1% of sugar in peptone water
along with an appropriate indicator. Durham’s tube is
kept inverted in tube to detect gas production
Eg:-Hiss serum sugars
• Transport media:-In the case of delicate organisms
which may not survive the time taken for transporting
specimen, special media are devised for transporting.
Eg:-Stuart’s medium

12. 3. Aerobic and anaerobic media
• Aerobic media are used to grow aerobes
Eg:-Mannitol salt agar
• Anaerobic media is used to grow anaerobic
organisms
Eg:-Robertson’s cooked meat agar

13. Culture method are of two types
Aerobic culture methods
Anaerobic culture methods

14. AEROBIC CULTURE METHODS
• 1. Streak plate
The streak culture (surface
plating) method
Is routinely used for the
isolation of bacteria in
Pure culture from clinical
specimens

16. 2.Lawn culture
• The lawn or carpet culture provides uniform surface
growth of the bacterium and is useful for
bacteriophage typing and antibiotic sensitivity
testing

18. 3.Stroke culture
• It is made in tubes containing agar slopes and
is employed for providing a pure growth of the
bacterium for slide agglutination and other
diagnostic purposes

19. 4.Stab cultures
These are prepared by
puncturing a suitable medium
such as nutrient gelatin or
glucose agar with a long, straight,
charged wire.
It used for demonstration of
gelatin liquefaction, oxygen
requirements of the bacterium
under study and maintenance of
stock culture

20. 5.Pour plate culture

22. 6.Liquid cultures
• Liquid cultures in tubes, bottles or flasks may
be inoculated by touching with a charged loop
or by adding the inoculum with pipettes or
syringes

23. ANAEROBIC CULTURE METHODS
• 1.McIntosh- Filde’s anaerobic jar

24. 2.Gaspak
This is now the
method of choice for
anaerobiosis.
The gaspak is
commercially
available as a
disposable envelope
containing chemicals
which generate
hydrogen and carbon
dioxide on addition
of water

26. MICROBIAL NUTRITION
o All chemicals required by micro organisms are
raw materials for the their metabolism and
reproduction are called NUTRIENTS
o All such events that help fulfilling the
requirements of nutrients to and into the
body of micro organisms are referred to as
NUTRITION

27. NUTRITIONAL REQUIREMENTS
MINERAL NUTRIENTS GROWTH FACTORS
MACRO NUTRIENTS
MICRO NUTRIENTS

28. MINERAL NUTRIENTS
MACRO OR MAJOR NUTRIENTS
• OXYGEN
• HYDROGEN
• CARBON
• NITROGEN
• PHOSPHOROUS
• SULPHUR
• POTTASSIUM
• MAGNESIUM
• SODIUM
• CALCIUM
• IRON
MICRO OR MINOR NUTRIENTS
• COBALT
• COPPER
• MANGANESE
• MOLYBDNUM
• NICKEL
• SELINIUM
• TUNGSTEN
• VANADIUM
• ZINC

29. MACRO NUTRINTS
• CARBON:- Main constituent of all organic cell
materials, CO₂ is the most oxidized form of
carbon
• SULPHUR AND NITROGEN:-Are subsequently
reduced within the cell and utilized in other
biosynthetic process
• MOLECULAR OXYGEN:-It need for respiration,
serves as terminal electron acceptor
• HYDROGEN:-Constituent of cellular water, organic
cell materials

30. • PHOSPHOROUS:-Constituent of nucleic acids,
phospholipids, and co-enzymes
• POTTASSIUM:-Important inorganic cations in
cells, co-factor for enzymatic reactions
• SODIUM:-Important inorganic cations in cells, and
have a role in membrane transport
• CALCIUM:-Inorganic cations, found in endospores
• MAGNESIUM:-Involved in protein synthesis
• IRON:-Constituent of cytochromes and other
haeme proteins

31. MICRO NUTRIENTS
• COBALT:-Vitamin B₁₂, transcarboxylase
• COPPER:-Respiration, photosynthesis
• MANGANESE:-Activator of various enzymes
• MOLYBDNUM:-Present in nitrogenase
• NICKEL:-Present in hydrogenase, urease
• SELINIUM & TUNGSTON:-In some formate
dehydrogenase
• VANADIUM:-Vanadium nitrogenase
• ZINC:-In DNA and RNA polymerases, many DNA
binding proteins

32. GROWTH FACTORS
AMINOACIDS PURINES
AND
PYRAMIDINES
VITAMINS

33. VITAMINS
• RIBOFLAVIN:-Precursor of FMN and FAD
• COBALAMINE:-Synthesis of deoxyribose
• BIOTIN:-In fatty acid biosynthesis
• PABA:-Precursor of folic acid
• FOLIC ACID:-Transfer of methyl groups
• THIAMINE:-Transketolase
• NIACIN:-Precursor of NAD
• LIPOIC ACID:-Acyl group transfer
• PANTHOTHENIC ACID:-Precursor of Co-A
• VITAMIN B₆:-Amino acid and keto acid transformation
• VITAMIN K:-Electron transport
• HYDROXAMATES:-Solubilization of Iron and transport
into the cell

35. CABBON
SOURCE
• AUTOTROPHS- CO₂ is the sole or principal source
• HETEROTROPHS-Reduced, preformed, organic molecule from
other organisms
ENERGY
SOURCE
• PHOTOTROPHS-Light
• CHEMOTROPHS-Oxidation of organic or inorganic
compounds
ELECTRON
SOURCE
• LITHOTROPHS-Reduced inorganic molecule
• ORGANOTROPHS-Organic molecules

36. ophs
TYPES
Photolitho
autotrophs
Chemolitho
autotrophs
Photoorgano
heterotrophs
Chemo
organo
heterotrop-
hs

37. PHOTOLITHO AUTOTROPHS
• Light chlorophyll energy
O₂
H₂O
H₂
CO₂ carbohydrates
Eg:-Green sulphur bacteria, Purple sulphur
bacteria

38. CHEMOLITHO AUTOTROPHS
• Inorganic metabolites Inorganic byproducts
H₂O
CO₂ Carbohydrates
Eg:-Sulphur oxidising bacteria, Iron oxidising
bacteria
O
H₂
ENERGY

39. PHOTO ORGANO HETEROTROPHS
• Light Bacteriochlorophyll Energy
Organic compounds
CO₂ Carbohydrates
Eg:-Purple non-sulphur bacteria
Organic byproducts
H₂

40. CHEMO ORGANO HETEROTROPHS
• They obtain food directly from external
environment using chemical energy source
• Eg:-Protozoa, Fungi
Chemo
organotrophy
HOLOTROPHIC NUTRITION ABSORPTIVE NUTRITION
Phagocytosis Pinocytosis

41. HOLOTROPHIC
• This form of nutrition is accomplished by free
living protozoa and slime molds which directly
engulf food
1. Phagocytosis:-Uptake of solid objects through
small invaginations in the cell membrane
2. Pinocytosis:-Uptake of fluids and soluble
nutrients through small invaginations in cell
membrane
ABSORPTIVE
• Obtain their food from external environment in
dissolved form by the process of absoption
Eg:-Bacteria, Fungi