Skin mucus is a non-lethal and low-invasive matrix appropriate to assess fish welfare as it contributes to their defence against external aggressions and reflects changes in fish health status. However, more information on the response of this matrix to specific stressors is needed. In this study, rainbow trout (Oncorhynchus mykiss) specimens were subjected to an acute stress by air exposure and sampled after 1, 6, and 24 h post-stress. Blood and skin mucus were collected, and a battery of biochemical biomarkers were measured in both matrices. Cortisol and glucose values showed the expected classical stress response in plasma, increasing after the acute stress. The same pattern was observed in skin mucus, corroborating previous data in fish, and allowing us to confirm that skin mucus can be a useful complementary matrix for stress assessment in fish. The results showed sensitivity to hypoxic stress in skin mucus for cortisol, glucose, alkaline phosphatase (ALP), aspartate transaminase (AST), alanine aminotransferase (ALT), creatinine kinase (CK), and calcium. From the 15 parameters evaluated, 12 did not show statistically significant changes between plasma and mucus; therefore, using skin mucus cannot replace the use of plasma. Finally, the principal component analysis in skin mucus revealed a complete separation between the two experimental groups, being ALP, AST, glucose, cortisol, and CK, the biomarkers that contributed the most to this separation.

Nanomaterials can adsorb biomolecules to their surface and form a protein corona. Here we investigated the protein profile bound to different sizes of aminated and carboxylated polystyrene (PS) nanoparticles after passing through the digestive tract of the freshwater zooplankter Daphnia magna. We found that acutely toxic aminated 53 nm PS nanoparticles bind a different set of proteins compared to other non-toxic PS nanoparticles. The aminated PS nanoparticles bind a higher number of proteins, which are smaller and more acidic, compared to the proteins which bind to the PS nanoparticles that are non-toxic in acute toxicity tests. The proteins bound to toxic nanoparticles can be divided into two groups. One group of proteins which function is related to the digestive system, whereas the other group of proteins can be related to the epithelium, intracellular structures and processes. Finally, we observed that not only proteins bind to the surfaces of the nanoparticles. Triglycerides effectively bind to 200 nm carboxylated PS nanoparticles but not to the other tested nanoparticles. These results provide information about the composition of the corona formed on surfaces of nanoparticles after a short-term incubation with D. magna and give insights to what underlies the acute toxicity caused by nanoparticles.

Nanoplastics (NPs) can cross cellular membranes and affect cellular performance. This study aims to determine the effects of polystyrene NPs (PS-NPs, 44 nm) on gilthead seabream (Sparus aurata) exposed for 14 days to 100 μg/L PS-NPs. The results show that biometric indicators (weight, length, Fulton’s condition factor, and hepatosomatic index) were not affected after the experimental exposures. No significant effects were observed on white blood cell count, red blood cell count, mean corpuscular hemoglobin, or platelets compared to the control group. However, there was a significant decrease in hemoglobin concentration, hematocrit values, and mean corpuscular cell volume in fish exposed to PS-NPs. There were no significant effects on plasmatic cholesterol, triglyceride, alkaline phosphatase, or aspartate aminotransferase levels. The histological anatomy of both the gills and the intestine revealed no obvious signs of cellular damage, excessive mucous, or inflammation in the PS-NP group. The expression of transcripts related to lipid metabolism (pparα, pparβ), growth and development (igf1), detoxification (bche), and oxidative stress (sod, gpx1) were significantly downregulated in animals exposed to PS-NPs, indicating a slight impairment in lipid homeostasis and antioxidant response.

Hematological studies provide essential information about the health of animals, which is crucial for veterinary medicine, scientific research, and aquaculture. Automatic hematological analyzers are an alternative to manual methods, offering faster and more reliable results. The objective of this study was to validate the Sysmex XN-1000V automatic hematology analyzer for blood samples from rainbow trout (Oncorhynchus mykiss), examine the effects of two anticoagulants (K₂EDTA and lithium heparin), and establish normal blood reference values for this fish species. Additionally, comparative studies were conducted between the Sysmex XN-1000V and manual methods (hemocytometer cell count and blood smear estimation), and reference intervals were established. Ninety-nine heparinized blood samples were analyzed for validation and sample stability tests. The results showed extremely good precision, with a coefficient of variation (CV) below 3% for RBCs, HGB, and HCT and less than 5% for non-RBC cells (leukocytes plus thrombocytes). However, heterophils (%) exhibited higher variability, with a CV of 15.08%. Linearity was excellent, and the carry-over was below 1% for all parameters. The sample stability test indicated that samples could be analyzed for up to 48 h when stored at 4 °C and up to 24 h at room temperature. Non-RBC cells were the first to degrade over time. The automated and manual methods demonstrated good correlation and agreement, validating the analyzer’s accuracy. The effects of two anticoagulants, K₂EDTA and lithium heparin, on the blood samples were also studied. Heparin was the preferred anticoagulant for routine hematological analysis of rainbow trout blood with the Sysmex XN-1000V analyzer. In conclusion, the Sysmex XN-1000V enables complete hemogram analyses to be performed quickly and accurately, standardizing techniques, harmonizing results, and providing reliable reference intervals with O mykiss blood.

The high incidence of anthropogenic impacts in the Mediterranean basin raises concerns on the health and quality of commercial fish species. This study aims to evaluate the health status of the European hake, Merluccius merluccius, from three areas of the Catalan coast (NW Mediterranean Sea) with different anthropogenic impacts (i.e., chemical pollution, litter, …) and assess if hake could serve as a sentinel species. We measured biomarkers of chemical exposure including B-esterases, antioxidant enzymes (GST, GR, GPx, CAT), biotransformation markers (EROD), lipid peroxidation, and macro-parasite assemblages. Hake showed, generally, a good health status across all areas with homogeneous patterns for most parameters. Tissue-specific differences included elevated gonadal cholinesterases and higher brain and hepatic carboxylesterase activities in the south, and increased hepatic EROD but lower lipid peroxidation in the central Barcelona area. Parasite assemblages were dominated by Digenea, Cestoda, and Nematoda, with higher cestode prevalence in both central and south zones. In summary, despite a greater prevalence of environmental pollution in the central region, there was a homogeneous pattern in hake health indicators throughout the three studied fishing zones. These results establish a baseline for hake health in Mediterranean waters and suggest that the species’ high mobility and wide depth range may limit its utility to detect local-scale pollution impacts, though it may serve as a regional-scale bioindicator.