Resumo. Abstract: Aim In this study, we investigated the distribution of Anabaenopsis elenkinii and Limnospira platensis in the saline-alkaline lakes of Nhecolândia (Pantanal wetland) and evaluated the impact of pH, temperature, and nitrogen on their growth and development to understand their ecological responses, showing insights into their ecophysiology in both cultured and natural environments. Methods Both species were collected in the subsurface, using a plastic bottle (200 mL) and the parameters temperature, conductivity, and pH were measured in situ. From these samples, the strains A. elenkinii CCIBt1059 and L. platensis CCIBt3335 were isolated and underwent six different cultivation treatments, in triplicate, during 30 days, with daily cell count, photoperiod of 12-12 hours of light-dark, and light intensity between 80-100 µmol photons m-2.s-1, using BG-11 modified medium, as follows: nitrate concentration experiments were performed with a 750 mg.L-1 NaNO3 (50%) and a nitrogen-free condition (0%) (T1 vs T2); temperature experiments were performed with 30 °C and 35 °C (T3 vs T4); pH experiments with 10.5 and 7.0 (T5 vs T6). The adopted control conditions were BG-11m medium (3% NaNO3, 45 mg.L-1), pH adjusted to 9.5, and temperature set at 25 °C. Results We have found that the treatment with the highest nitrogen availability (T1), pH of 9.5, and a temperature of 25 °C, provides the most favorable conditions for the growth of both studied species. In nature, A. elenkinii occurred predominantly between pH 9.04 to 10.4 (average 9.8) and L. platensis at pH 9.22 to 10.23 (mean 9.9), highlighting the alkaliphilic nature of these species. Furthermore, we observed that temperature influences the frequency of heterocyte formation in A. elenkinii. At elevated temperatures (30 and 35 °C), the frequency of heterocytes was higher compared to 25 °C during the exponential growth phase, indicating that increased heterocyte formation is a strategy in response to temperature stress. Conclusions This research provides valuable insights into the ecological aspects and optimization of the cultivation of the two species studied, which hold ecological significance to saline lakes. Further studies are recommended to explore their potential biotechnological applications.