Sandy Biochem (1)

ISOLATION OF PROTEINS IN GUT MICROBES FROM
Oreochromis niloticus (TILAPIA FISH)
Guide Name Submitted By

Dr.K.Jayanthi Santhiya B
Assistant Professor P22BN009
Department of Microbiology and Biotechnology II M.Sc., Biochemistry
Faculty of Arts and Science Department of Microbiology and Biotechnology Faculty
Bharath Institute of Higher Education and Research of Arts and Science
Selaiyur, Chennai Bharath Institute of Higher Education and Research
Selaiyur, Chennai
INTRODUCTION
Human beings consume a significant number of pathogens every day,
primarily bacteria. Probiotic bacteria have been used in various diets for a
number of decades because of their beneficial effects on human health.
The Agriculture Organization of the United States and the World
Health Organization. Defined probiotics in 2001 as bacteria that, when given
to a host at adequate levels, improve their health (Bonifait et al., 2009).

Live bacteria and yeasts are known as probiotics, and they may be good for one's
health. They can be found in some meals and supplements as well as in the human
digestive system. Probiotic bacteria are beneficial. They are found all over the body,
although most people only think of the stomach and intestines when they think of them.
In many researchers findings infections of the digestive tract, irritable bowel,
lactose intolerance, allergies, infections of the urogenital tract, cystic fibrosis, and various
cancers can all be prevented and treated with the use of probiotics ( Bodke et al., 2022).
AIM AND OBJECTIVES
Aim
Isolation of protein from mapped probiotic bacteria present in gut

microbiome of Oreochromis niloticus.
Objectives
 To collect and dissection of the Oreochromis niloticus (Tilapia fish) samples.
 To isolate the mapped protein from Oreochromis niloticus sample gut.
 To analysis of microbial culture techniques (Plating, colonies identification).
 Characterization analysis by Ion Exchange Chromatography (IEC) Techniques.

METHODOLOGY
Collection of Oreochromis niloticus (Tilapia fish) and dissection of the samples
Dissection of Oreochromis niloticus
Gut isolation
Colonies identification
Microbial culture
Characterization analysis by Ion Exchange Chromatography (IEC) Techniques.

PROGRESS REPORT
Fig.1: Dissection of Fish Sample Fig.2: Isolation of gut microbes by spread plate technique
Fig.3: Identification of gut microbes by streak plate technique Fig.4: Isolation of DNA by Agarose Gel Electrophoresis
method
BIFIDOBACTERIUM (GRAM POSITIVE)
BIOCHEMICAL TEST
• Biochemical tests help in determining the specific species present, which is crucial for accurate
bacterial identification.
• I performed the test that includes
• Indole Test,
• Methyl Red Test,
• Voges Proskauer Test,
• Urease Test,
• Citrate Utilization Test,
• Triple Iron Test,
• Oxidase Test,
• Catalase Test.
INDOLE TEST:
The test is important in the identification of Enterobacteria.
Most strains of species break down the amino acid tryptophan with the release of indole.
Negative Reaction : No color change even after the addition of appropriate reagent.
METHYL RED TEST:
The test detects the production of sufficient acid during the fermentation of glucose and the maintenance of
conditions such that the pH.
The test bacteria is grown in broth medium containing glucose.
Positive Reaction : A distinct red color is formed.
VOGES-PROSKAUER TEST:
The test is used to determine if an organism produces acetylmethyl carbinol from glucose fermentation.
Negative Reaction: A lack of a pink red color.
CITRATE UTILIZATION TEST:
Citrate agar is used to test an organism’s ability to utilize citrate as a source of energy.
Positive Reaction: Growth with color change from green to intense blue along the slant.
TRIPLE SUGAR IRON TEST:
The test is used to determine the ability of an organism to ferment glucose, lactose, and sucrose, and their ability
to produce hydrogen sulfide.
An alkaline/acid (red slant/yellow butt) reaction with gas production: It is indicative of dextrose fermentation only
UREASE TEST:
It is also known as the CLO test(Campylobacter-like organism test)
Negative Reaction: No color change
CATALASE TEST:
If catalase is very active due to an abundance of hydrogen peroxide, the rapid production of oxygen
gas(O2) will produce bubbles.
Positive Reaction: It is indicated by bubbling
OXIDASE TEST:
The oxidase test detects the presence of a cytochrome oxidase system that will catalyse the
transport of electrons between electron donors in the bacteria .
Dry filter Paper Method using Positive Reaction: Intense deep blue, Negative Reaction: Absence of
coloration.
Fig 2.OXIDASE TEST(Negative)

Fig 1.CATALASE TEST(Positive)
Fig3 : A B C D E F
Fig3: (A) Indole Test (Negative), (B) Methyl Red Test(Positive), (C) Voges Proskauer Test(Negative), (D)
Citrate Utilization Test(Positive), (E) Triple Iron Test(red slant with yellow butt and gas production, (F) Urease
Test(Negative)

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ISOLATION OF PROTEINS IN GUT MICROBES FROM

Oreochromis niloticus (TILAPIA FISH)

Guide Name Submitted By

Human beings consume a significant number of pathogens every day,

number of decades because of their beneficial effects on human health.

The Agriculture Organization of the United States and the World

Health Organization. Defined probiotics in 2001 as bacteria that, when given

to a host at adequate levels, improve their health (Bonifait et al., 2009).

In many researchers findings infections of the digestive tract, irritable bowel,

Isolation of protein from mapped probiotic bacteria present in gut

 To isolate the mapped protein from Oreochromis niloticus sample gut.

 To analysis of microbial culture techniques (Plating, colonies identification).

 Characterization analysis by Ion Exchange Chromatography (IEC) Techniques.

Dissection of Oreochromis niloticus

Characterization analysis by Ion Exchange Chromatography (IEC) Techniques.

Fig 2.OXIDASE TEST(Negative)

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