1. Introduction Conclusion References Aim of the work
First identification of Erugosquilla massavensis from Mediterranean Sea, Egypt using DNA barcoding technique
Amira T. Abo-Hashesh1, Wafaa S. Sallam 2, Fedekar F. Madkour 3, Amro M. S. Hanora 4, Hanaa K. Ashour5
1Institute of Biotechnology for Postgraduate and Research, Suez Canal University, Egypt 2Department of Marine Science, Faculty of Science, Suez Canal University, Egypt 3Department of Marine Science, Faculty of Science, Port Said University, Egypt 4Department of Microbiology, Faculty of Pharmacy, Suez Canal University, Egypt 5Department of Chemistry, Faculty of Science, Suez Canal University, Egypt
Results & Discussion
Introduction
E. massavensis was identified by using information of barcoding gene region.
Phylogenetic analysis represented the sequence of E. massavensis appear in different branch of the other squillidae species, confirmed that studied sequence is belonging to family squillidae and approved that amplified sequence of E. massavensis was not submitted in gene bank database before (Fig. 7).
Squilla is one of the economic marine crustaceans in the Mediterranean Sea. It is cheap and rich food source. Also, It considers as a good source for chitin and chitosan production.
Genetic identification studies are less focusing on crustaceans as in family Squillidae and no genomic sequence of Erugosquilla massavensis were found in the genetic database.
DNA barcoding technology, using short sequences of the mitochondrial gene cytochrome oxidase subunit 1 (COI), has been proposed as a method for enabling rapid and accurate detection and identification of species.
Crustacean COI barcode region exhibited the highest species-level divergence rate among all animal groups.
Fig. (4): Genomic DNA extracted from E. massavensis using QIAamp mini kit (Qiagene, Germany)
Aim of the work
This study is aimed to record the first identification of E. massavensis inhabiting Mediterranean Sea, Egypt using DNA barcoding technique.
Feed gene bank database with a new sequence of one careless family (Squillidae).
Materials & Methods
E. massavensis samples were collected from the Mediterranean Sea, Port Said, Egypt (Fig.1).
Morphological identification of E. massavensis specimens were carried by Manning (1995).
Molecular processes steps and tools described in Figures (2 and 3).
Fig. (6): DNA sequence of partial amplified COI gene of E. massavensis obtained from DNA sequencer (3500 genetic analyzer; Applied Biosystems, USA).
Fig. (5): Partial amplified COI of E. massavensis
Fig. 1: Map of sampling area; the Egyptian Mediterranean Sea at Port Said, Egypt.
Fig. (7): Phylogenetic tree of partial coding COI gene sequence of E. massavensis was compared with other sequences of family Squillidae from gene bank using MEGA 6 program and M. monoceros as a out group.
Conclusion
Fig. (2): Molecular processes steps of DNA extraction of E. massavensis and PCR amplification of COI gene using the primer pair; CrustF1 (forward) and HCO2198 (reverse) described by Costa et al. (2007). DNA extracted and PCR product were visualized by horizontal gel electrophoresis. DNA sequencing and analysis for amplified partial COI gene.
First identification of E. massavensis from Mediterranean Sea using DNA barcoding technique was achieved.
A new sequence was added to gene bank database.
Using of DNA barcoding technique of the mitochondrial COI gene proved to be a good technique in identification.
More molecular identifications were needed to be submitted in the gene bank database.
Acknowledgements
Our praiseful gratitude is expressed to Biotechnology Institute for Postgraduate and Research, Suez canal University, Egypt for providing facility to doing this work in their labs.
References
Costa F. O., deWaard J. R., Boutillier J., Ratnasingham S., Dooh R. T., Hajibabaei M., and Hebert P. D.N. (2007). Biological identifications through DNA barcodes: the case of the Crustacea. Canadian J of Fish and Aqua Scie.
Manning R. B. (1995). Stomatopod crustacea of Vietnam: the legacy of Raou1 Serene. Crust. Res. 4:  
Fig. (3): Tools were used in DNA extraction (QIAamp mini kit), PCR amplification (Mastercycler gradient PCR; eppendorf, Germany), gel electrophoresis and DNA sequencer (3500 genetic analyzer; Applied Biosystems, USA) and analysis for amplified partial COI gene using NCBI database and MEGA 6 program.