## 2 Materials and Methods
### Crude Lipids  
The total lipids content was gravimetrically quantified following a continuous extraction process with diethyl ether in a Soxhlet apparatus (Behr Labor-Technik GmbH, Dusseldorf, Germany), as it follows the international standard AOAC method 930.09 [34]. The distillation flasks were previously dried at 105 \({}^{\circ}\)C for 2 h, cooled in a desiccator, and weighed using an analytical scale (Sartorix, Gottingen, Germany). Afterwards, the distillation flasks were filled (to 2/3 of their capacity) with diethyl ether (Panreac, Darmstadt, Germany). Then, approximately 2 g of the algal samples (Sartorix, Gottingen, Germany) were packed in filter paper and placed into the thimble. After 16 h of extraction, all the solvent was collected and evaporated (BUCHI Labortechnik AG, Flawil, Switzerland). The distillation flasks were then dried at 105 \({}^{\circ}\)C for 2 h and weighed (Sartorix, Germany) when cooled down.  
Crude lipids were calculated according to the formula presented by the standard method of AOAC 930.09 [34]:  
\[Crude\;lipids\;(\%\;db)=100\times\frac{P3-P1}{P2}\]  
\[Crude\;lipids\;(\%\;fb)=\frac{Crude\;lipids\;(\%\;db)\times(100-H)}{100}\]  
\(\%\;db\)--percentage of dried biomass; \(\%\;fb\)--percentage of fresh biomass; \(P1\)--distillation flask weight (g); \(P2\)--sample weight (g); \(P3\)--distillation flask weight + lipids (g); \(H\)--moisture (\(\%\)).  
Fatty acids were extracted from dry algal biomass and trans-methylated to fatty acid methyl esters (FAMEs) for analysis as described by [PERSON] et al. (2012) [35]. Samples were incubated with methanol (Fisher Chemical, Waltham, MA, USA) for the extraction of lipids. The nonadecanoic acid (C19:0) (Fluka 74208) was added as an internal standard for further quantification. Samples were centrifuged (Sorvall(tm) ST16, Thermo Scientific, Waltham, MA, USA) for 25 min at 5 \({}^{\circ}\)C, 537 g, and stored in liquid form at \(-80\)\({}^{\circ}\)C until further analysis.  
FAMEs' identification was done by gas chromatography-mass spectrometry (GC-MS) using Thermo Scientific Trace 1310 Network (Waltham, MA, USA) equipment, equipped with a TR-FFAP column (Thermo Scientific, Waltham, MA, USA) of 0.32 mm internal diameter, 0.25 \(\upmu\)m film thickness, and 30 m long. The sample (0.60 \(\upmu\)L) was injected in splitless mode, at an injector temperature of 250 \({}^{\circ}\)C, lined with a split glass liner of 4.0 mm i.d. The initial oven temperature was 80 \({}^{\circ}\)C, following a linear temperature increase of 25 \({}^{\circ}\)C min\({}^{-1}\) to 160 \({}^{\circ}\)C, followed by another ramp of 2 \({}^{\circ}\)C min\({}^{-1}\) to 210 \({}^{\circ}\)C, and finally an increase of 40 \({}^{\circ}\)C min\({}^{-1}\) until a final temperature of 230 \({}^{\circ}\)C was reached and maintained for 10 min. Helium at a flow rate of 1.4 mL min\({}^{-1}\) was used as a carrier gas. A Thermo Scientific ISQ 7000 Network Mass Selective Detector (Waltham, MA, USA) at scanning m/zranges specific for fatty acids in Selected Ion Monitoring (SIM) mode acquisition was used. The detector starts operating 3.5 min after injection, which corresponds to solvent delay. The injector ion source and transfer line were maintained at 240 \({}^{\circ}\)C and 230 \({}^{\circ}\)C, respectively. Integration of FAME peaks were carried out using the equipment's software Xcalibur (tm) (Thermo Scientific, Waltham, MA, USA). Identification of each peak was performed by retention time and mass spectrum of each FAME, comparing to the Supelco(r) 37 component FAME mix (Sigma-Aldrich, Steinheim, Germany). Quantification of FAMEs was done as described in [PERSON] et al. (2012) [35].