Description: Milkfish (Chanos chanos) is one of the most important aquaculture species in Asian countries. These teleost fish are traditionally cultured in outdoor-based systems and therefore have to cope with daily and/or seasonally changing environmental conditions. Temperature changes beyond the optimal range of a fish species are known to induce an endocrine stress response resulting in the release of cortisol via the hypothalamic-pituitary-interrenal axis. Moreover, (thermal) stress induces glucocorticoid-mediated changes in the fish's energy metabolism to cope with the stressor(s) and regain homeostasis. Long-term elevations of cortisol are known to be detrimental for fish performance. In this study, we investigated the stress response of juvenile milkfish, which were exposed to a gradual temperature increase of 1°C per day over 7 days in the range from 26°C to 33°C, followed by an exposure to constant 33°C for 21 days. We quantified ontogenetic (OG) and regenerated (RG) scale cortisol to evaluate chronic stress. To investigate metabolic implications and oxidative stress response, activity levels of key enzymes involved in metabolic (isocitrate dehydrogenase - IDH, lactate dehydrogenase - LDH, electron transfer system - ETS) and antioxidant (superoxide dismutase - SOD, catalase - CAT) related pathways were quantified. Furthermore, we measured available energy resources (protein, carbohydrates, lipids) and potential cellular damage due to oxidative stress (lipid peroxidation - LPO). Finally, changes in the gut microbiome of the milkfish related to the temperature stress were analyzed to elucidate their role in the stress response and interactions with physiological parameters. This study is part of the ACUTE project (AquaCUlture practice in Tropical coastal Ecosystems - Understanding ecological and socio-economic consequences) funded by the Leibniz Association grant SAW-2015-ZMT-4. It is associated with the following publications: Hanke et al., 2019 (doi:10.1016/j.aquaculture.2018.09.016) and Hassenrück et al., 2020 (doi:10.3390/microorganisms9010005). The final OTU table and statistical analysis scripts for Hassenrück et al., 2020 are supplied as further details to this data set.

Description: Upwelling systems are significant sources of atmospheric nitrous oxide (N₂O). The Benguela Upwelling System is one of the most productive regions worldwide and a temporally variable source of N₂O. Strong O₂ depletions above the shelf are favoring periodically OMZ formations. We aimed to assess underlying N₂O production and consumption processes on different temporal and spatial scales during austral winter in the Benguela Upwelling System, when O₂-deficiency in the water column is relatively low. The fieldwork took place during the cruise M157 (August 4th – September 16th 2019) onboard the R/V METEOR. This expedition included four close-coastal regions around Walvis Bay at 23°S, which presented the lowest O₂ concentrations near the seafloor and thus may provide hotspots of N₂O production. Seawater was collected in 10 L free-flow bottles by using a rosette system equipped with conductivity-temperature-depth (CTD) sensors (SBE 911plus, Seabird-electronics, USA). Incubation experiments were performed using stable isotope ¹⁵N-tracers. Seawater samples for ¹⁵N-tracer incubations and natural abundance N₂O analysis were collected from 10 L free-flow bottles and filled bubble-free into 125 mL serum bottles. The samples for natural abundance N₂O analysis were immediately fixed with saturated HgCl₂ and stored in the dark. To perform the incubation, we added ¹⁵N-labeled NO₂-, NO₃⁻ and NH₄⁺ to estimate the in-situ N₂O production rates and associated reactions. To determine a single rate, the bottles were sacrificed after tracer addition, and within the time interval of 12 h, 24 h and 48 h by adding HgCl₂. Rates were calculated based on a linear regression over time. Total N₂O and natural abundance isotopologues of N₂O were analyzed by using an isotope ratio mass spectrometer (IRMS, Delta V Plus, Thermo Scientific). NO₂- production was additionally analyzed by transforming ¹⁵NO₂- to ¹⁵N₂O following the azide method after McIlvin & Altabet (2005) and the nitrogen isotope ratio of N₂O was measured by an IRMS. N₂ production was determined via an IRMS (Flash-EA-ConfloIV-DELTA V Advanced, Thermo Scientific) by injecting headspace from exetainers. The N₂O yield per nitrite produced and the N₂O yield during denitrification was calculated. Samples for natural abundance N₂O was sampled and measured in triplicates and is shown as an average with standard deviation (SD). In order to estimate the contribution of different N₂O producing pathways by major biological processes and the extent of N₂O reduction to N₂, the dual-isotope mapping approach was applied to natural abundance isotopologues of N₂O, which uses the relative position of background-subtracted N₂O samples in a δ¹⁵Nˢᴾ-N₂O vs. δ¹⁸O-N₂O diagram (Yu et al., 2020; Lewicka-Szczebak et al., 2020).