National institute of oceanography Laboratory of genetics

Genotyping of Plankton Useful for

Feeding Common Egyptian Marine

Fishes and their Fries

البصمة الوراثية لبعض الهائمات المفيدة لتغذية معظم األسماك

. البحرية المصرية وزريعتها

Presentation By

Name: Hany Tolba

Supervisors

Prof. Dr Mohamed Abd Elsalam Rashed professor of Genetics, Faculty of Agriculture Ain Shams university

Prof. Dr Mahmod Ahmed Abd Elhafez Sallam professor of Genetics, Faculty of Agriculture Ain Shams university

Dr Hosam Elsaied Assistant professor of Genetics, NIOF

Introduction

What are marine common fishes in Egypt ?

Of course

Mugilidae - Mugil cephalus - Liza ramada - Liza aurata

Mugil cephalus Liza aurata Liza ramada

The idea of the study :Problems in feeding

Mullet fish ?

Traditional methodology

No complete profile for Wild Mullet gut Microbiome

Disadvantages Most of Microbiome

can not be detected

undre microscope ,

because , they are

partially digested .

Mullet gut

Genomics Technique

Looking microscopy

Main Targets of the study

1- Obtain a complete Phylogenetic for Wild type gut Microbiom

2- Construct Life Gene Atlas, for Plankton Feeding Wild Mullet.

What is the Microbime Types Expected to be Found in Gut of Mullet?

Bacteria Archaea Eukaryote

red slime algae ( cyanobacteria)

Blue Green algae ( cyanobacteria)

Crab Larva Copepod

Crab Larva

Oerbacteriën (Methanopyrus kandleri)

1- Sampling of wild mullet and evacuation of gut content

2- Metagenomic DNA Extraction of gut microbiome

3- PCR amplification of small subunit rRNA gene as the finger print of gut microbial species

4- Denaturant Gradient Gel Electrophoresis, DGGE

5- Analysis of DGGE product by sequencing

6- Construct phylogenetic trees of recorded species based on sequenced rRNA genes

Plan of Work: for Detection of Microbiome in the gut of Mullet.

1- Sampling of wild mullet and evacuation of gut content

Bardwell Lake Manzala Lake

Fishing

Mullet Gut

Preparation of gut content of Mullet

Evacuation

Fry

Hosam Elsaied research group

2- Metagenomic DNA Extraction of gut microbiome

This is an Example for DNA Extraction of Gut Content from 5 Fish Individual .

Hosam Elsaied research group

3-PCR Using rRNA gene

Specific Primers

Bacteria Archaea Eukaryote

16 S rRNA gene

Types of amplified r RNA gene

18 S r RNA gene

rRNA Genes

4- Denaturant Gradient Gel Electrophoresis, DGGE

Mechanism of DGGE

5- Analysis of DGGE product by sequencing

Example of use DGGE Technique for eukaryote sample

Common plankton rRNA gene band occurred in both samples

a b 100 bp ladder

100 bp

Polyacrylamide DGGE gel

Agarose gel

Applied to polyacrylamide DGGE as duplicate samples

Analyzed by sequence

PCR using 18S rRNA gene of Metagenomic DNA ( a , b )

Hosam Elsaied research group

6- Construct phylogenetic tree of recorded species based on sequenced rRNA genes

PCR Using rRNA gene(16S,18S) by Specific Primers

Environmental sample (mullet gut content)

Metagenomic DNA Extraction

DGGE

sequencing

phylogenetic

Summary

thanks