The aim of the study was to explore the possibility of bioremediation of oil refinery wastewaters by the cyanobacterium Synechococcus sp. MK568070, isolated from the Adriatic Sea. The potential of biomass and lipid production was explored upon cultivation on oil refinery wastewater with excess CO₂ after the removal of nutrients. The strain grew well in a wide range of salinities and ammonium concentrations, and was further tested on the wastewater from local oil refinery plant of various N-composition. Growth experiment under optimized conditions was used to analyze the lipid, carbohydrate and protein dynamics. The biomass yield was highly dependent on nutrient source and concentration, salinity and CO₂ addition. Highest biomass yield was 767 mg/L of dry weight. Towards the end of the experiment the decline in carbohydrate to 18.9% is visible, whereas at the same point lipids, in particular saturated fatty acid methyl esters (FAME), started to accumulate within the cells. The content of lipids at the end of the experiment was 21.4%, with the unsaturation index 0.45 providing good biofuel feedstock characteristics. Fourier Transform Infrared (FTIR) spectroscopy analysis demonstrated a high degree of lipid accumulation in respect to proteins, along with the structural changes and biomass accumulation. In addition, the N-removal from the wastewater was >99% efficient. The potential of lipid accumulation, due to the functional photosynthesis even at the minimal cell quota of nutrients, is critical for the usage of excess industrial CO₂ and its industrial transformation to biodiesel. These findings enable further considerations of Synechococcus sp. (MK568070) for the industrial scale biomass production and wastewater remediation.

Three metagenomic libraries were constructed using surface sediment samples from the northern Adriatic Sea. Two of the samples were taken from a highly polluted and an unpolluted site respectively. The third sample from a polluted site had been enriched using crude oil. The results of the metagenome analyses were incorporated in the REDPET relational database (http://redpet.bioinfo.pbf.hr/REDPET), which was generated using the previously developed MEGGASENSE platform. The database includes taxonomic data to allow the assessment of the biodiversity of metagenomic libraries and a general functional analysis of genes using hidden Markov model (HMM) profiles based on the KEGG database. A set of 22 specialised HMM profiles was developed to detect putative genes for hydrocarbon-degrading enzymes. Use of these profiles showed that the metagenomic library generated after selection on crude oil had enriched genes for aerobic n-alkane degradation. The use of this system for bioprospecting was exemplified using potential alkB and almA genes from this library.

The microalgae of the genus Pseudochloris/Picochlorum are characterized by fast growth, and wide nutrient (type and concentration) and salinity tolerance, all contributing towards exploration of their use in high-density biomass production and wastewater bioremediation. In this study, removal of nitrogen and phosphorus nutrients from oil refinery wastewater was monitored during growth of the marine eukaryotic microalgae Pseudochloris wilhelmii, with emphasis on biochemical analyses of its biomass quality to evaluate suitability for biodiesel production. A series of growth experiments under various nutrient and light regimes were performed in a temperature range of 20-30°C to evaluate nutrient removal and biomass growth dependence on temperature. The highest removal rate of dissolved inorganic nitrogen reached under the given experimental conditions was 0.823 mmol/(gday) accompanied by the corresponding biomass productivity of 115.2 mg/(Lday). Depending on light and temperature, the final lipid concentration ranged 181.5 – 319.8 mg/L. Furthermore, increase in nutrient load decreased the maximum specific growth rate by 25%, and the maximum specific removal rate of the dissolved inorganic nitrogen by 19%, whereas the duration of bioremediation process was nearly doubled. In contrast, constant light exposure expedited the nitrogen removal, i.e. bioremediation process, by almost 40%, while supporting over three times higher biomass productivity and the highest maximum specific growth rate of 0.528 g/(gday). The conditions favoring the highest nitrogen removal and highest toxicity reduction in oil refinery wastewater are met at 24°C and 130 µmol phot/(m²s). The highest proportion of carbon-binding to the P. wilhelmii biomass was noticed under the same conditions, thus indicating them as the most favorable conditions for hydrocarbon removal as well as for CO₂ sequestration. Pseudochloris wilhelmii therefore represents a promising candidate for oil refinery wastewater remediation and valuable biomass cogeneration on a large-scale.

Microalage are broadly recognized as promising agents for sustainable wastewater treatment and biomass generation. However, industrial effluents such as petroleum refinery wastewater (WW) present challenges due to toxic growth inhibiting substances. Three marine microalgae species: Pseudochloris wilhelmii, Nannochloropsis gaditana and Synechococcus sp. MK568070 were examined for cultivation potential in oil refinery WW. Their performance was evaluated in terms of growth dynamics, lipid productivity, and toxicity reduction, with a focus on their suitability for largescale industrial use. N. gaditana demonstrated the highest growth rate and lipid content (37% d.w.) as well as lipid productivity (29.45 mg/(Lday)) with the N-uptake rate of 0.698 mmol/(gday). The highest specific DIN uptake rate was observed inn P. wilhelmii (0.895 mmol/(gday) along with the highest volumetric productivity (93.9 mg/L/day) and WW toxicity removal (76.5%), while Synechococcus sp. MK568070 demonstrated lower performance metrics. A simple numerical model was applied to calculate continuous operation based on empirical results of batch experiments. Sustainability of the microalgae-based WW remediation under the conditions of optimized lipid biomass production was estimated, regarding 2019–2022–2025 cost dynamics. Parameters for optimum open raceway pond cultivation were calculated, and the biomass production accumulation was estimated, with the highest biomass production noted in P. wilhelmii (171.38 t/year). Comparison of treatment costs, production costs and revenue showed that the best candidate for WW remediation is N. gaditana.

The aim of the study was to evaluate the growth, fatty acid profile and concentration of malondialdehyde of muscle tissue of meagre Argyrosomus regius fed diets with different lipid content. The long-term experiment was conducted in three feeding groups: A (CP = 52.0; CL = 21.0), B (CP = 56.0; CL = 18.0), C (CP = 48.0; CL = 16.0) with two replicates in marine net cages on Bisage Island, Adriatic Sea over 20 months. At the beginning of the experiment, fish were of equal weight (6.83 ± 1.03 g) and length (8.57 ± 0.49 cm) and were fed to satiation during the experiment. At the end of the experiment, the fish from each feeding group (n = 110) were measured and muscle tissue was collected (n = 60) and stored at −80 °C until analysis. The final weight and condition factor were significantly different (p < 0.05) between the groups. The highest ratio of crude fats and n-3/n-6-fatty acids was found in the muscle tissue of group A. Fish fed diet A also exhibited higher MDA levels compared to fish in the other feeding groups, indicating elevated levels of lipid peroxidation in muscle tissues. Experimental feeding group A showed better growth performance, a higher content of the beneficial fatty acids EPA and DHA and a more favorable n-3/n-6 ratio than feeding groups B and C. Continuously monitoring and adjusting feeding protocols in accordance with lipid content and fatty acid composition could maximize growth and health outcomes in meagre farming.

The aim of the study was to explore the relationship between membrane-regulating functional lipids of three fucacean species and their associated epiphytic bacterial communities. The analyzed algae Treptacantha barbata, Carpodesmia crinita and Cystoseira compressa, formerly classified under the Cystoseira genus, are indigenous to the Adriatic Sea. The thalli of sampled Fucales species were divided into perennial and annual parts. T. barbata and C. crinita show high contents of mono- and polyunsaturated fatty acids but, however, show a clear distinction between branches, cauloids and apical parts. Along with the highest unsaturation degree (4.32), the C. crinita branches and apical parts demonstrated high proportion of both C18 and C20 polyunsaturated fatty acids, with eicosapentaenoic (C20:5 n-3) and arachidonic (C20:4 n-6) fatty acids up to 5.46% and 13.82%, respectively. The highest proportion of saturated fatty acids was found in thalli of C. compressa, particularly on cauloids and old branches (≥90%). The similarity profile routine analysis of fatty acids and microbial community structure has shown clear separation of the cauloids and apical parts from the branches of C. crinita and T. barbata. Based on the nonmetric multidimensional scaling analysis several representatives of the Gammaproteobacteria class, identified as Vibrionales (Vibrio), Cellvibrionales and Xanthomonadales, which contributed strongly to the separation of T. barbata apical parts and branches, C. compressa receptacles and C. crinita branches from the T. barbata cauloids and C. crinita apical parts and cauloids. The highest richness in polyunsaturated fatty acids, in particular C18:3(n-3), C20:4(n-6) and C20:5(n-3), was observed on branches, accompanied by a distinctive epiphytic microbial structure dominated by numerous representatives with potentially beneficial biological activity. The results showed a strong relationship between fatty acid profiles of the analyzed species and phycosphere community structure, underlining the host physiological condition in shaping the biological interactions and maintaining a healthy microbiome, as well as compiling the ecophysiological and molecular research in order to better assess the resilience of the ecosystem.