ALBERT

Description: This study examined the metabolic response of juvenile turbot (Scophthalmus maximus) to diets with graded fishmeal (FM) replacement with plant, animal, and emerging protein sources (PLANT, PAP, and MIX) in comparison to a commercial-like diet (CTRL). The feeding experiment was carried out from April to July 2019 in the Centre for Aquaculture Research (ZAF) at the Alfred Wegener Institute for Polar and Marine research in Bremerhaven, Germany. The juvenile turbot (Scophthalmus maximus) were purchased from France Turbot (L'Épine, France) and acclimated to the recirculating aquaculture system (RAS) for 2 weeks prior to starting the 16 weeks experimental trial. To elucidate the effects of the protein sources and the level of FM replacement on the metabolic response of the fish, a 1H‐nuclear magnetic resonance (NMR) spectroscopy was used to assess the metabolic profiles of muscle and liver tissue after feeding the fish the experimental diets for 16 weeks. Feed, muscle, and liver samples were ground under liquid nitrogen and approx. 200–250 mg tissue was homogenized in 5x volume of ice‐cold 0.6 M perchloric acid (PCA) (w:v). After one cycle of 20 s at 6000 rpm and 3 °C, using Precellys 24 (Bertin Technologies, Montigny‐le‐Bretonneux, France), samples were sonicated for 2 min at 0 °C and 360 W (Branson Sonifier 450, FisherScientific, Schwerte, Germany). Homogenates of the experimental diets, muscle and liver tissues were centrifuged for 2 min at 0 °C and 16,000 g, and supernatants were neutralized with ice cold potassium hydroxide (KOH) and PCA to pH 7.0–7.5. To remove precipitated potassium, perchlorate samples were centrifuged again for 2 min at 0 °C and 16,000 g. The entire supernatant was transferred, shock‐ frozen in liquid nitrogen, and stored an −80 °C for later analysis. One‐dimensional 1H‐NMR spectra for feed and tissues extracts were acquired using a vertical 9.4 T wide bore magnet with Avance III HD (Bruker‐GmbH, Ettlingen, Germany) at 400.13 MHz with a 1.7 mm diameter triple tuned (1H‐13C‐15N) probe. Each spectrum was processed and analyzed with Chenomx NMR Suite 8.4 software (Chenomx Inc., Edmonton, Canada). Before analyzing, the spectra were corrected for phase, shim and baseline and calibrated to trymethylsilyl proprionate (TSP) signal (at 0.0 ppm).

Description: Raw data acquired by an SBE21 thermosalinograph and an auxiliary SBE38 temperature sensor (Sea-Bird Scientific, USA) installed in an underway seawater flow-through system on board RV Heincke were processed to yield a calibrated and validated data set of seawater temperature and salinity along the cruise track. The seawater inlet is located at a depth of 2 m. The raw hexadecimal data were downloaded from the DAVIS SHIP data base (https://dship.awi.de) at a resolution of 1 s, and converted to temperature and conductivity using the pre-deployment factory calibration coefficients. The converted data were averaged to 1 min values, outliers were removed, and sensor drift was corrected using coefficients obtained from a post-season calibration performed at Sea-Bird at the end of the measurement season. Salinity was calculated from internal temperature, conductivity and pressure according to the PSS-78 Practical Salinity Scale. Processed data are provided as 1 min means of seawater temperature, conductivity and salinity, aligned with position data taken from the master track. Quality flags are appended according to the SeaDataNet Data Quality Control Procedures (version from May 2010). More details are described in the attached processing report.