The study about Nile Pufferfish (Tetraodon lineatus) and its growth conditions in Lake Nasser, Aswan, Egypt which is still at a very early stage. Recently, Pufferfish might be considered nutritious food due to its content of good amount of oil, protein, minerals. A total of 30 Nile Pufferfish samples were collected using commercial nets from Lake Nasser (15 fish samples from Khor El Ramla as a North Lake and 15 samples from Tushka East as a South Lake). The total lengths of North and south Lake samples ranged between 16 – 39 and 16 - 28cm with a mean of 27.1 and 21.3 cm, respectively. While, the total body weight of North and south Lake samples ranged between 136 – 1636 and 129-608 g with a mean of 680.8 and 265.7 g, respectively. In addition, the results indicate that there are different groups from T. lineatus of the two locations based on the partial sequence of CO1 gene. The nucleotide frequencies of CO1 gene of North Lake samples were 25.80%, 31.45%, 24.75% and 18.00% for T, C, A and G, respectively. While, the Nucleotide frequencies of South Lake samples were 25.96%, 31.62%, 24.31% and 18.11% for T, C, A and G, respectively. The habitat and location of fish has significant effects on the weight and length of Pufferfish but the origin has the same father. Segregation was happened in the deep water which leads to more haplotypes in the same area. Therefore, proper utilization of this species towards sustainable management, nutritional composition, and biosecurity issues will lead to achieving a sustainable blue economy.