Population's biological parameters, including length at first capture, mortalities, exploitation rates, growth coefficient, longevity, and recruitment times, are essential in assessing fishing status, but there is no data on Mystus mysticetus. Therefore, the study was conducted to provide these parameters to assess the fishing status of this species at Cai Rang, Can Tho (CRCT) and Long Phu, Soc Trang (LPST). A collection of 741 individual fish was used for analysis and showed that most fish size groups ranged from 9.0 cm to 12.0 cm, and the asymptotic length was 16.8 cm for both CRCT and LPST populations. The fish population von Bertalanffy curve was Lt = 16.80(1 − e−0.51(t + 0.38)) at CRCT and Lt = 16.80(1 − e−0.48(t + 0.40)) at LPST. The fish growth coefficient at CRCT (2.16) was higher than at LPST (2.13), whereas the reverse case was true for longevity ranging from 5.88 years (at CRCT) to 6.25 years (at LPST). At CRCT, fishing mortality, natural mortality, total mortality, and exploitation rate were 0.69/ year, 1.40/year, 2.09/year, and 0.33, respectively; at LPST, these values were 0.75/ year, 1.33/year, 2.08/year, and 0.36, respectively. Although the population parameter of this fish species exhibited a spatial variation, both CRCT and LPST fish resources have not been subjected to overexploit because E (0.33 at CRCT and 0.36 at LPST) is lower than E 0.1 (0.707 at CRCT and 0.616 at LPST).