TOPIC :

SUGAR FERMENTATION TEST

PRESENTED BY : RAJA AADIL REG. NO. : AAH MA02 - 01

INTRODUCTION

Fermentation is a metabolic process performed by many types of bacteria

Fermentation allows the production of ATP without the need for atmospheric oxygen

Many different kinds of fermentable carbohydrates

Not all bacteria can ferment all of these carbohydrates One commonality among fermentation is the production of acids

AIM
To determine the ability of a microorganisms to ferment a specific carbohydrate producing acid or acid with visible gas

PRINCIPLE

Bacteria grown in a liquid culture medium containing the carbohydrate

They may produce organic acids as by-products of the fermentation

These acids are released in to the medium and so lower pH of medium Phenol red or bromo cresol blue detects the change in pH Gases produced can be detected by durhams tube

Materials Required

Phenol Red Carbohydrate Fermentation Broth Bacterial culture. Inoculation loop. Incubator(370 C)

MEDIA USED
Phenol-Red Carbohydrate Fermentation Broths are useful for helping to characterize bacteria based on their fermentation abilities

COMPOSITION:
Single fermentable carbohydrate: 0.5g Trypticase: 1g Phenol-Red..RED at pH 7.4 : 0.0189mg Sodium Chloride: 0.5g

PROCEDURE
Preparation of Carbohydrate Fermentation Broth

Weigh and dissolve trypticase, Sodium chloride, and Phenol red in 100 ml distilled water and transfer into conical flasks. Add 0.5% to 1% of desired carbohydrate into all flask Distribute 5 ml of broth in sterile test tubes Insert inverted Durham tubes into all tubes Sterilize at 1150 C for 15 minutes

CONTS
Inoculation of Bacterial Culture into the Phenol Red

Carbohydrate Broth

Aseptically inoculate each labeled carbohydrate broth with bacterial culture Incubate the tubes at 18-24 hours at 37oC Observe the reaction

RESULTS

Acid production:
Changes the medium into yellow colour

Acid and Gas production:

Changes the medium into yellow colour Gas production can be detected by the presence of small bubbles in the inverted durham tubes

Absence of fermentation:
The broth retains the red color