Identifikasi Lactobacillus & Bifidobacterium

Lactobacillus

• Sampel : Feses 10 gr

• Wadah : Screw Cap Steril

• Prosedur : 1. Feses dihomogenkan dengan NaCl 0.9 /

Akuades steril

2. Dilakukan kultur pada media MRS Broth 1 : 9

3. Dilakukan isolasi LB pada MRS agar

4. Identifikasi koloni Lactobacillus

Proses inkubasi 72 h/ 37 C, Suasana Anaerob

MRS broth (DeMan, Rogosa, Sharpe Broth)

MRS BrothComposition per liter:• Glucose ....................................................................................... 18.5g• Pancreatic digest of gelatin ......................................................... 10.0g• Beef extract ................................................................................... 8.0g• Yeast extract.................................................................................. 4.0g• Sodium acetate.............................................................................. 3.0g• K2HPO4......................................................................................... 2.0g• Ammonium citrate ........................................................................ 2.0g• Polysorbate 80............................................................................... 1.0g• MgSO4·7H2O................................................................................ 0.2g• MnSO4·4H2O.............................................................................. 0.05gpH 6.2 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from BD Diagnostic Systems.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the isolation and cultivation of Lactobacillus species from clinical specimens, foods, and dairy products.

Bifidobacterium

• Sampel : Feses 10 gr

• Wadah : Screw Cap Steril

• Prosedur : • Feses dihomogenkan dengan NaCl 0.9 /

Akuades steril• Dilakukan kultur pada media TYP Broth 1 : 9• Dilakukan isolasi LB pada TYP agar• Identifikasi koloni Bifidobacterium

Proses inkubasi 72 h/ 37 C, Suasana Anaerob

TPY BrothTPY MediumComposition per liter:• Pancreatic digest of casein.......................................................... 10.0g• Glucose ......................................................................................... 5.0g• Pancreatic digest of soybean meal ................................................ 5.0g• Yeast extract.................................................................................. 2.5g• K2HPO4......................................................................................... 2.0g• Cysteine·HCl................................................................................. 0.5g• MgCl2·6H2O.................................................................................. 0.5g• ZnSO4·7H2O ............................................................................... 0.25g• CaCl2........................................................................................... 0.15g• FeCl3 ...........................................................................................1.0μg• Tween™ 80................................................................................1.0mL• pH 6.5 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat until dissolved. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the isolation and cultivation of Bifidobacterium species.