RESUMEN
Extreme weather events, like marine heatwaves (MHWs), are becoming more frequent and severe due to climate change, posing several challenges to marine ecosystems and their services. As disease outbreaks are often prompted by these acute phenomena, it is essential to develop eco-innovative strategies that can efficiently improve farmed fish resilience, especially under sub-optimal rearing conditions, thereby ensuring a sustainable aquaculture production. This study aimed to unveil farmed juvenile white seabream (Diplodus sargus, 28.50 ± 1.10 g weight, n = 150) immune and antioxidant responses under a category II MHW in the Mediterranean Sea (+4 °C, 8 days of temperature increase plus 15 days of plateau at the peak temperature) and to investigate whether a 30 days period of prophylactic biofortification with Asparagopsis taxiformis (1.5 %, 3 % and 6 %) enhanced fish resilience to these extreme events. Several biomarkers from different organization levels (individual, cellular, biochemical and molecular) were assessed upon 30 days of biofortification (T30), exposure (after 8 days of temperature increase + 15 days at peak temperature, T53) and recovery (8 days of temperature decrease, T61) from the MHW. Results showed that MHW negatively affected the fish physiological status and overall well-being, decreasing specific growth rate (SGR) and haematocrit (Ht) and increasing erythrocyte nuclear abnormalities (ENAs) and lipid peroxidation (LPO). These adverse effects were alleviated through biofortification with A. taxiformis. Seaweed inclusion at 1.5 % was the most effective dose to minimize the severity of MHW effects, significantly improving immune responses of D. sargus (i.e. increased levels of immunoglobulin M, peroxidase activity and lysozyme expression) and modulating antioxidant responses (i.e. decreased LPO, catalase and glutathione S-transferase activity). These findings confirm that A. taxiformis is a functional ingredient of added value to the aquaculture industry, as its inclusion in marine fish diets can beneficially modulate fish immunity and resilience under optimal and adverse rearing conditions.
RESUMEN
BACKGROUND: The forskolin-induced swelling (FIS) assay measures CFTR function on patient-derived intestinal organoids (PDIOs) and may guide treatment selection for individuals with Cystic Fibrosis (CF). The aim of this study is to demonstrate the repeatability and reproducibility of the FIS assay following a detailed Standard Operating Procedure (SOP), thus advancing the validation of the assay for precision medicine (theranostic) applications. METHODS: Over a 2-year period, FIS responses to CFTR modulators were measured in four European labs. PDIOs from six subjects with CF carrying different CFTR genotypes were used to assess the repeatability and reproducibility across the dynamic range of the assay. RESULTS: Technical, intra-assay repeatability was high (Lin's concordance correlation coefficient (CCC) 0.95-0.98). Experimental, within-subject repeatability was also high within each lab (CCCs all >0.9). Longer-term repeatability (>1 year) showed more variability (CCCs from 0.67 to 0.95). The reproducibility between labs was also high (CCC ranging from 0.92 to 0.97). Exploratory analysis also found that between-lab percentage of agreement of dichotomized CFTR modulator outcomes for predefined FIS thresholds ranged between 78 and 100 %. CONCLUSIONS: The observed repeatability and reproducibility of the FIS assay within and across different labs is high and support the use of FIS as biomarker of CFTR function in the presence or absence of CFTR modulators.