LAB 5_Bacteria cultivation: culture media, pure culture

OBJECTIVES:
1. The purpuse of bacterial cultivation
2. The types of culture media
3. The procedure of cultivation in 4 sectors in order to obtain the pure culture
1. The purpuse of bacterial cultivation
Medical
- the etiological diagnosis of infectious diseases
- susceptibility testing at antibiotics
- the necesity of obtaining pure culture
The pure culture contains bateria from the same type.
Industrial obtaining of the antigens, vacines, biosynthesis products.
2. Definition
The culture media: watery solution which contains nutritive substaces (nutrients and growth factors).
Fastidious bacteria are dependent by many growth factors.
The required conditin for one culture media:
- to be nutritive
- to have the osmotic pressure and pH at the same values like internal medium.
- to be sterile
- to be transparent
The physical chemical conditions useful for cultivation:
- Osmotic pressure: - the majority of pathogenic bacteria grow in isotonic media with the internal medium of the
host
- halophylic bacteria ( >2 % NaCl)
- pH - the majority of bacteria needs one Ph around (7-7,5)
- acidophylic bacteria (5-5,5) / basophylic bacteria (8,5-9)
- temperature:
- psihrotrofic bacteria: fast multiplication at 20-30C and slow cultivation at the fridge temperature
- psihrofylic bacteria (0-20C)
- mezophyilic bacteria (25-45C)
- termophylic bacteria (50-70C).
- atmosfere of incubation:
- strictly aerobic bacteria = can grow only in the presence of oxigen
- strictly anaerobic bacteria = can grow only in the absence of oxigen
- aerobic facultative anaerobic = cultivate aerobic and anaerobic
- microaerophylic bacteria = cultivate only in the presence of low oxigen concentration (5%)
- carboxiphylic bacteria = require for growing CO2 concentration bigger than those from the air (510%)
Culture media classification:
a. regarding the sourse of nutrients:
- empirical animal, vegetal source
- synthetical pure chemical substances
b. regarding the nutritive values:
-simple non fastidious bacteria (e.g., agar)
- enriched fastidious bacteria (e.g., blood agar)
c. regarding the consistency:
- liquids monobacterial pathologic products
- solids pluribacterial pathologic products
- semisolids allow the study of mobility
d. Regarding the purpuse of use:
- isolation
uzual (e.g., blood agar)
differentials (e.g., blood agar, Mac Conkey)
selective (Mac Conkey, Hektoen)
enrichment (selenit broth)
- identification (MIU, TSI, Simmons)
- preservation

4. The cultivation and isolation in pure culture

The usual technique for bacterial cultivation for obtaining the pure culture involve cultivation in 4 sectors.

Culture = all the bacteria accumulated by multiplying

Colonie = culture, visible with naked eye, from one single bacteria multiplication named CFU colonies forming
units.
Colonie = one bacterial colonie, so it is one pure culture.