Photosynthetic microorganisms, such as microalgae, are remarkable for their ability to harness sunlight, fix carbon dioxide, and produce a variety of bioactive compounds. These organisms are pivotal in climate mitigation strategies as they can absorb carbon dioxide while generating valuable biomolecules. Among the diverse cultivation approaches, mixotrophic growth combines light energy with both inorganic and organic carbon sources, offering a unique strategy to enhance biomass production and metabolic diversity in microalgae. Here, microalgal species such as Nannochloropsis granulata, Phaeodactylum tricornutum, and Chlorella sp. were investigated for their potential applications under different cultivation methods, including phototrophy and mixotrophy. Mixotrophic conditions significantly improved biomass production across all tested species. Among these, Phaeodactylum tricornutum, a marine diatom, emerged as a promising candidate for bioactive compound production, exhibiting higher antiproliferative activity against human melanoma cells and antibacterial effects against Staphylococcus aureus. Importantly, Chlorella sp. was also found to possess antibacterial activity against Staphylococcus aureus, broadening its potential applications. Additionally, metabolomics analysis was performed on Chlorella sp. and Phaeodactylum tricornutum to identify the compounds responsible for the observed bioactivity. This study highlights the value of mixotrophic cultivation in enhancing the productivity and bioactivity of microalgae, positioning them as versatile organisms for sustainable biotechnological applications.

The salt ponds of Trapani, Sicily, represent an extreme and under‐explored ecosystem characterised by varying salinity gradients and environmental conditions. These ponds, integral to traditional salt extraction, include cold, driving, hot and crystallizer ponds, each hosting diverse microbial communities. This study aimed to explore the biological and physical–chemical diversity of 11 ponds during the salt production season in Trapani. We conducted comprehensive physical–chemical characterizations, including measurements of pH, conductivity, viscosity, density, organic carbon and ion concentration. Microbial DNA was extracted from salt pond waters and subjected to metabarcoding of 16S rRNA genes to determine the diversity of archaea and bacteria. High‐throughput sequencing revealed significant variations in microbial communities across different pond types and seasons. Cold ponds showed a higher diversity of moderately halophilic organisms, while crystallizer and feeding ponds were dominated by extreme halophiles, particularly archaeal genus Halorubrum and Haloquadratum and bacterial genus Salinibacter . Statistical analyses indicated that environmental parameters, especially salinity and temperature, significantly influenced microbial community composition. Our findings enhance the understanding of microbial ecology in saline environments and highlight the potential of halophilic microorganisms. This study provides a foundation for future research into the functional roles of these microorganisms and their industrial applications.