This study aims to genotype plankton found in the guts of common Egyptian marine fish and their fry. Researchers will extract metagenomic DNA from the gut contents of wild mullet to obtain a complete phylogenetic profile of their gut microbiome. They will use PCR and denaturing gradient gel electrophoresis (DGGE) to amplify and analyze small subunit rRNA genes from the bacteria, archaea, and eukaryotes present. Sequencing the rRNA gene bands will allow construction of phylogenetic trees to identify the microbial species in the mullet guts. This genotyping technique provides a more complete picture of the wild mullet gut microbiome compared to traditional microscopy methods.

1. National institute of oceanography
Laboratory of genetics
Genotyping of Plankton Useful for
Feeding Common Egyptian Marine
Fishes and their Fries
‫األسماك‬ ‫معظم‬ ‫لتغذية‬ ‫المفيدة‬ ‫الهائمات‬ ‫لبعض‬ ‫الوراثية‬ ‫البصمة‬
‫وزريعتها‬ ‫المصرية‬ ‫البحرية‬.
Presentation
By
Name: Hany Tolba

2. 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

3. Introduction

4. What are marine common fishes in Egypt ?
Of course
Mugilidae
- Mugil cephalus
- Liza ramada
- Liza aurata
Mugil cephalus Liza aurataLiza ramada

5. 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

6. 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.

7. 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 LarvaCopepod
Crab Larva
Oerbacteriën
(Methanopyrus
kandleri)

8. 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.

9. 1- Sampling of wild mullet and
evacuation of gut content
Bardwell LakeManzala Lake

10. Fishing

11. Mullet Gut
Preparation of gut content of Mullet
Evacuation
Fry
Hosam Elsaied research group

12. 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

13. 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

14. rRNA Genes

15. 4- Denaturant Gradient Gel Electrophoresis, DGGE

16. Mechanism of DGGE

17. 5- Analysis of DGGE product by sequencing

18. 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

19. 6- Construct phylogenetic tree of recorded
species based on sequenced rRNA genes

20. PCR Using rRNA gene(16S,18S)
by Specific Primers
Environmental sample
(mullet gut content)
Metagenomic DNA Extraction
DGGE
sequencing
phylogenetic
Summary

21. thanks