Sugar Fermentation Test

7/28/2019 Sugar Fermentation Test

1/15

SUGAR FERMENTATION TEST

PRINCIPLE:

Fermentations are energy producing biochemical reactions

in which organic molecules severs as both electron acceptor

and electron donors. The ability of microorganisms toferment the carbohydrate the types of formed product are

very useful in heir identification. The given carbohydrate

may be converted into a number of different end product

depending upon the microorganisms involved. The product

may be acid gases alcohols are the characteristics of the

that particular organism.

Test culture suspension was taken from enriched

broth

Loopful culture was inoculated in peptone

water base containing 1% respective sugar

Incubate on shaker for 24 hrs at 37 c

Observe the result and note down


2/15

IMVIC TESTS

INDOLE TEST

Principle:The amino acid tryptophan is found in nearly all

proteins. Bacteria that contain the tryptophanase canhydrolyze tryptophan to its metabolic products,

namely, Indole, pyruvic acid,and ammonia. The

bacteria use the pyruvic acid and ammonia to satis fy

nutritional needs; Indol is not used and accumulates

in the medium. The presence of indol can be detected

by the addition of Kovacs reagent. Kovacs reagent

reacts with the indol, producing a bright redcompound on the surface of medium.

Tryptophanindol, pyruvate and ammonia.

Test culture suspension prepared in sterile saline

Loopful suspension is inoculated in sterile peptone

water base


3/15

All the tubes incubated at 37c for 24 hrs

Results observed and noted

CONFIRMATORY TEST

1. BILE TEST

Inoculate the loopful inoculum in the bile medium

(MRS + bile2% & 3%)

Incubate over night on shaker at 37c for 24 hrs.

Check the turbidity in the medium

The tubes which shows turbidity will carry

cell adhesion


4/15

Inoculate the loopful enriched broth in the

mediumhaving pH1 & pH 3

Incubate the medium overnight on shaker at

37c for 24 hrs

Check the turbidity

The tubes which shows turbidity will carry for

cell adhesion


5/15

OXIDATIVE FERMENTATIVE TEST

Prepare the medium

Inoculate the culture with the help of stab wire

In the oxidative test only inoculum is added but in

fermentative test, as soon as inoculum added,paraffin oil added to cover the surface.

Check the color change and gas formation

in the both tubes


6/15

ANTIBACTERIAL ACTIVITY

Prepare nutrient agar

Spread the sensitive organism on the plate

Plates were incubated in refrigerator for 10 min

Make the well with the help of borer

Inoculate the enriched broth in specific well

After incubation, measure the zone of inhibition


7/15

NITRATE REDUCTION TEST

Chemolithoautotrophic bacteria and many

Chemoorganoheterotrophs can use nitrate as a

terminal electron acceptor during anaerobic

respiration. In this processes, nitrate is reduced to

nitrite by nitrate reductase.

Test culture suspension prepared in sterile

saline

Loopful suspension is inoculated in sterile

peptone water base

All the tubes incubated at 37c for 24 hrs

Results observed and noted


8/15

SUGAR FERMENTATION TEST

Fermentations are energy producing biochemical

reactions in which organic molecules severs as both

electron acceptor and electron donors. The ability of

microorganisms to ferment the carbohydrate the

types of formed product are very useful in their

identification. the given carbohydrate may be

converted into a number of different end productdepending upon the microorganisms involved. the

product may be acid gases , alcohols depending on

the characteristics of the that particular organism.

Test culture suspension was taken from

enriched broth

Loopful culture was inoculated in peptone

water base containing 1% respective sugar


9/15

Incubate on shaker for 24 hrs at 37 c

Observe the results and note down

OXIDASE TEST

Oxidase enzyme plays important role in electrontransport system. The production of the oxidase

enzyme can be checked by using reagent N,

N,N,N-Tetra methyl -P-Phenylendiamine

dihydrochlride

A small quantity of bacteria is taken on filter

paper disc soaked in reagent. In positive test, color

of dye get changed.


10/15

CATALASE TEST

Some bacteria contains flavoprotien that reduces

O2 resulting production of hydrogen peroxide

which is extremely toxic. Survival in the presence

of antimetabolite is possible due to enzyme called

as catalase.

H2O22H2O + O2

Inoculate loopful test culture

Dipped into catalase reagent

Observe effervesces in positive test


11/15

CERTIFICATE

This is to certify that Mrs. Ashwini A pensalwarstudying in M.Sc. Microbiology, has worked on a

project entitled during

the period of in a partial

fulfillment of the requirement for Masters degree in

microbiology, is record of candidates own work

carried out by her under supervision and guidance.

The matter embodied this report has not been

submitted for award for any other degree.


12/15

DELERATION

The project entitled has been submitted

to university of pune as a partial fulfillment M.Sc degree

under the guidance of Dr. suneeti Gore (Department of

microbiology, Fergusson college pune) and literature cited

has been written in my own words.


13/15

ASHWINI A.PENSALWAR

M.SC. MICROBIOLOGY

FERGUSSON COLLEGE,

PUNE

Contents: Page

number

Objective

Abstract

Introduction

Material and Methods

Results

Discussion

References


14/15

Appendix

Nutrient Agar

Peptone - 1gm

Meat extract -0.3

Sodium chloride-0.5Agar -2%

D/W -100ml

pH -6.8

Peptone water base

Peptone

Sodium chloride

D/W


15/15

Documents

Sugar Fermentation Test