1

(Bacterial Culture Bacterial culture media)

I.
II.
III. (Cultural Characteristics )
IV. (Bacterial Culture Media)
V. (Preparation of the Culture Media)
VI. (Maintenance of Bacteria)

I.

1. ( Energy )
- Phototroph
- ( Oxidation )
Chemotroph
2. (Carbon Sources)


Autotroph Photoautotroph
Chemoautotroph
Heterotroph

3. (Electron Sources)

Lithotroph
Organotroph
3. (Nitrogen sources)

4.

5.
K+, Mg2+,
Ca2+, Fe 2+ Co facter
6.

II.

Pure Culture
(Colony count)

1. ( Streak Plate )

(Colony)

Four - way cross Streak

Loop (Solid Media)
1 Loop


2

1 Streak Plate (Quinn et al., 1994)

2 Streak Plate ( , 2541)

2. ( Spread Plate )



spread plate 3
( petri dish turnable ) ( 4 )

3 Spread Plate ( , 2541 )

4 Petri dish turnable ( Cappuccino & Sherman , 1992 )

( Pour-plate Technique )
( 5)
45 C
(Serial dilution) Ten fold
Dilution ( 6)

(Incubate
Spread plate

7
5 (Pour-Plate) (Cappuccino & Sherman , 1992)

6 (Serial Dilution) (Cappuccino & Sherman,

1992)

III. ( cultural characteristics )

( 7)



1. (solid media)

1.1 ( Colony Size )

1.2 ( Colony form )
- Punctiform 1
- Circular
- Filamentous
- Irregular
- Rhizoid

1.3 ( Colony margin or Colony edge )

- Entire

- Undulate
- Lobate

- Erose

- Filamentous
- Curled

1.4 ( Surface Texture )

- Smooth

- Rough

- Mucoid

1.5 ( Elevation )
- Flat

- Raised

- Convex

- Pulvinate

1.6 ( Consistency )
- Butyrous
- Stringy

- Rubbery
1.7 ( Optical Feathures )
- Transparency
- Translucent
- Opaque

1.8 ( Chromogenesis or Pigmentation) Pyoverdin

Pseudomonas aeruginosa
2. ( Broth Media )
2.1
2.2
-
Turbid
Granular
-
Ring

Pellicle
Flocculent
-
Granular
Flocculent
Viscid

Flaky
3. ( Agar Slant )
3.1
3.2
- Filiform

- Echinulate
- Beaded

- Arborescent
- Rhizoid

- Effuse

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7 (Cappuccino & Sherman , 1992)

IV. (Bacterial Culture Media)

11

1.
2. ( Sterility test )
3.
4.
5. toxoid
6.
(Pathogen)
(Artificial Media) (Fastidious Bacteria)
Mycobacterium laprae

Growth factor Metabolism
(Sterilized)

heterotrophic microorganism
complex nitrogen compound
(crude natural protein source)


(protein
hydrolysates) peptones
Polypeptidase amino acids
Polypeptidase amino acids peptones

heterotrophic bacteria
peptones

(dehydrated media)

(Basic nutrient Media)

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1. Peptones : Peptone Amino

acid Simple Nitrogenous conpounds
peptones (
) Amino acids
Peptones Casein
( ) Gelatin Yeast cells Peptones

2. Infusion Extracts : ( Yeast
Extract Peptone ) ( Malt Extract)
( Beef Extract Brain- Heart infusion) Peptones
Peptones Infusion
extract
Polypeptides, aminoacids Growth factors
3. Solidifying agents :
(Agar) , Gelatin , Silica gel
Polyacrylic gels Polysacharides
(Rhodophyceae) 80 C
2% ( / )
Solidifying Agents
4. Indicators : Indicators 2
4.1 Indicators (pH)
Phenol red, Bromothymol Blue, Bromocresol Purple, Neutral red, litmus, andrade's
indicator
4.2 Indicators Oxidation- reduction potential (Eh)
Methylene Blue Resazurin
5. (Salt ;NaCl) Osmotic pressure
Isotonic

6. Dextrose Carbon
7.
(distilled water)

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8. Selective agent :
selective agent
selective agents
crystal violet brilliant green (
)
sodium chloride (
Staphylococcus spp.
Staphylococcus spp.), sodium azide, sodium citrate , sodium tellurite, sodium lauryl sulfate,
sodium Selenite, iodine, phenylethanol
( pH 5.6 abouraud dextrose agar)
( pH 8.8-9.0 alkaline peptone water Vibrio cholerae)
Selective Character
9. Reducing agent ;
ascobic acid , sodium thioglycolate ,
sodium formaldehyde sulfoxylate , thiomalic acid , sodium hydrosulfite , cysteine
10. ; Enriched Media 4

- Fibrin (defibrinated sheep blood) ;
hemolysis hemolysis Streptococci spp.
- Fibrin hemolysis

- fibrin hemolysis Streptococci

Mueller Hinton media
(lysed
horse blood)
- ;
citrate , antimicrobial agents
hemolysis

1.
1.1 (Solid Media) (Agar) 1.5 2 %

14

1.2 (Liquid Media or Broth)

0.1 %
1.3 (Semisolid Media)
0.5 %
(Motility test)
2. Selective Agents
2.1 Chemical Defined Media


2.2 Plain Media
Nutrient
Broth , Nutrient agar
2.3 Enriched Media (Fastidious
bacteria)
Growth Facter (Ascitic Fluid)
Blood Agar
Chocolate Agar
2.4 Enrichment Media
()
Selenite broth , tetrathionate broth
2.5 Selective Media

Crystal Violet Brilliant Green (bile salt)

Selective Media Brilliant green agar,
Salmonella Shigella agar (SS Agar) Bismuth sulfite agar MacConkey agar
2.6 Differential Media
(Biochemical Reaction)
Blood agar media

(Hemolysis) (Clear Zone) Lactose broth

15

2.7 Differential Selective media

Differential media Selective Media

Thiosulfate Citrate Bile salt Sucrose (TCBS) agar Vibrio
Vibrio Sucrose Sucrose
Xylose lysine Dexoxycholate Agar (XLD)

Salmonella
XLD (H2S)

2.8 (Assay Media)

(Disinfectant)
2.9 (Media for enumeration of bacteria)

Plate count agar Marine
agar
2.10
(Media for characterization of
bacteria)
(Physical and Biochemical test) Triple
sugar iron agar , Urease test medium , citrate agar , lysine decarboxylase test medium
2.11 (Maintenance Media)

Oxidation Reduction Potential
Stuarts transport medium , Cary Blair transport medium Transport
Media

16

V. (Preparation of the Culture Media)

(Dehydrated Media)
BBL ,Difco , Gibco Oxoid
Dehydrated Media (Granules)

1. (Glassware)
(Detergent)
2. Dehydrated media
Flask (Distilled Water)
(Chlorine)
(Heavy metal ions)
3.
2

4. Dehydrated media Dehydrated media

Hot plate
(Magnetic stirrer system) Flask
Dehydrated media
5. (Sterilized) Autoclave Pressure
cooker 121 C 15 15
118 C
115 C 15 Selective Media Brilliant green
agar , TCBS XLD
6. 50 56C
Water bath Plate Tube Agar 42C
1 Plate 70 100 Plates
7.
Autoclave
5056 C
(Aseptic technique)

17

8. (pH) (50 - 56 C)
pH meter pH strips
( 6.8 7.2) 1-N 0.1 N NaoH HCl
pH
9. Plate Tube
10. Plate Tube
37 C 2 3 Plate
Contamination Plate

4 . C
( Incubation ) 37 C 12 14

VI. (Maintenance of Bacteria)

1. (Subculturing Method)
Maintenance media

2.
(Agar slant Media)

1. (Lyophilization)

18

( Skim milk)
(Freeze - dry) 60 78 C


2.
196 C
()
Dimethyl sulfoxide , DMSO
10 30
Lyophilization

1. 2529 Medical Bacteriology

( MTMI 301 )
1 17 - 27

19

2. 2541

2
74 96
3. 2541
2
116 134
4. Baron, E.J., Pererson, L.R., Finegold, S.M.1998.Bailey and Scott's Diagnostic Microbiology.
10th ed. CV Mosby, St. Louis,
5. Cappucino,J.G. ,Sherman,N. 1992 . Microbiology a Laboratory Manual 3rd ed. The Benjamin /
Cummings Publishing Company, Inc. P. 63 - 81.
6. Koneman, E.W., Allen, S.D., Schreckenberg, P.C., Winn, W.C.1992.Atlas and Textbook of
Diagnostic Microbiology. 4th ed. JBLippincott, Philadelphia,
7. Murray, P.R., Baron, E.J., Pfaller, M.A., Tenover, P.C., Yolken, R.H. 1995.Manual of Clinical
Microbiology. 6th ed. American Society for Microbiology, Washington, DC.
8. Pratt, P.W. 1997. Diagnostic Microbiology. In: Laboratory Proceedures For Veterinary
Technicians. 3 rd. ed. Mosby Year Book ,Inc. P.119 - 258.
9. Quinn, P.J. Carter, M.E., Markey, B.K. and Carter, G.R. 1994. Clinical Veterinary
Microbiology.1st ed. Wolfe Publising, Mosby-Year Europe Limited, London, England, p.
6-345.