This study supplements the gender determination methodology based on the relationship of some morphological indicators in Mystus albolineatus. A total of 229 specimens were caught by trawl nets at Cai Rang – Can Tho and Long Phu – Soc Trang in four months from April to July 2023. After collection, the samples were transferred to the laboratory to determine the total length (TL), standard length (SL), eye diameter (ED), eye distance (DE), body height (BH), head length (HL), and mouth diameter (MD) before surgery to determine the sex based on the gonad. The results showed that the TL-SL regression relationship could be used to determine the sex of this fish. Besides, TL-BH and TL-MD growth relations were relatively balanced in females versus males. In addition, based on SL, DE, and MD, it was also possible to determine whether the sampling time was the wet or dry seasons. These findings form the basis for the initial study of the biological characteristics of this fish in the Mekong Delta.