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Bacterial nutrition and the design of culture media. Based on bacterial metabolism*Culture pHCulture oxidation- reduction petencialGaseous requirmentsOxzgen. Growth of bacteria. Growth of bacterial cellGrowth in batch culture. Growth in batch culture . The lag faseThe exponential fas
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1. Bacteria Growth in the laboratory
(in vitro)
2. Bacterial nutrition and the design of culture media Based on bacterial metabolism*
Culture pH
Culture oxidation- reduction petencial
Gaseous requirments
Oxzgen
3. Growth of bacteria Growth of bacterial cell
Growth in batch culture
4. Growth in batch culture The lag fase
The exponential fase
The stationary fase
5. Growth in batch culture
6. Bacterial growth on solid surface Agar media
Colony forming units
Bacterial colony
7. Environmetal conditions optimal temperature,
oxygen concentration,
pH,
water activity
8. Oxygen concentration Aerobs
Anaerobs (do not require oxygen)
Obligate anaerobs (die in the presence of O)
Facultative anaerobs (E.coli)
Microaerophilic bacteria
9. pH Acidophiles (grow at low pH (0-5,5)
Alcaliphiles (8,5-11,5)
Normal (6,5-7,2)
10. Temperature ( characteristic ranges) Psychrophiles: with optimum growth T around 20 C
Mesopihles: between 15 and 45 with optimum around 37 C
Thermophiles: between 30 and 75 with optimum around 55 C
Hyperthermophiles: T grater than 100C
11. Techniques used to study bacteria Aseptic (sterile) techniques:
Sterile media
To prevent contamination (accidental intorduction of unknown bacteria)
Sterilisation (autoclave, flaming)
Desinfection (the removal of potentially harmful microbes : B, V,
12. Baceria are grown (cultured) Growth media:
Liquid (for large numbers of bcteria)
Solid (for isolation of individual bacteria)
Semisolid ( for demonstration of motility)
Envinronmental conditions:
optimal temperature, oxygen concentration, pH,
water activity
13. Growth media Defined media (synthetic)- composed form defined ingredients
Complex media � composed from undefined ingredients such as proteolytic digests of meat (peptons) and meat extracts
Nutrient broth, tryptic soya broth
Nutrient agar,�
Blood- an addtive to media
14. Obtaining bacterial colonies Pure culture
Isolation � using method called streaking
To strake bacteria on to agar plates we are using a wire (or plastic) loop
15. Selective and differential media Selective media: for selection of particular groups of bacterial pathogens ( contain inhibitors i.e. antibiotics, bile salts, dyes, which are suppressing the growth of unwonted bacteria)
Differential media: for differentiation of two species or groups (lactose +, -)
16. Agar isolated from seaweed Is not degradated by bacteria
Agar is melted by boiling
Liquid medium can be converted into solid medium by the adition of agar (1- 2%) or semisolid medium (0,6%)
17. Colonies Shape
Size
Elevation
Edge
Surface
Opacity
Consistency
18. Counting of bacteria Viable counts (according of number of colonies)
Turbimetric measurements
Other methods (RT PCR)