Multi-marker DNA metabarcoding for precise species identification in ichthyoplankton samples.

Ichthyoplankton monitoring is crucial for stock assessments, offering insights into spawning grounds, stock size, seasons, recruitment, and changes in regional ichthyofauna. This study evaluates the efficiency of multi-marker DNA metabarcoding using mitochondrial cytochrome c oxidase subunit I (COI), 12S rRNA and 16S rRNA gene markers, in comparison to morphology-based methods for fish species identification in ichthyoplankton samples. Two transects with four coastal distance categories were sampled along the southern coast of Portugal, being each sample divided for molecular and morphological analyses. A total of 76 fish species were identified by both approaches, with DNA metabarcoding overperforming morphology-75 versus 11 species-level identifications. Linking species-level DNA identifications with higher taxonomic morphological identifications resolved several uncertainties associated with traditional methods. Multi-marker DNA metabarcoding improved fish species detection by 20-36% compared to using a single marker/amplicon, and identified 38 species in common, reinforcing the validity of our results. PERMANOVA analysis revealed significant differences in species communities based on the primer set employed, transect location, and distance from the coast. Our findings underscore the potential of DNA metabarcoding to assess ichthyoplankton diversity and suggest that its integration into routine surveys could enhance the accuracy and comprehensiveness of fish stock assessments.

Cerium uptake, translocation and toxicity in the salt marsh halophyte Halimione portulacoides (L.), Aellen.

Halimione portulacoides plants were exposed to dissolved cerium (Ce) in a hydroponic medium for five days. Ce accumulation in plants followed the metal's increase in the medium although with a very low translocation factor (TF < 0.01) between roots and shoots. Ce median concentrations in roots were 586, 988 and 1103 µg/g (dry wt.), while in shoots the median values reached 1.9, 3.5 and 10.0 µg/g (dry wt.), for plants exposed to 300, 600 and 1200 µg/L of Ce, respectively. No significant differences occurred in the length of roots and shoots among treatment groups, albeit plants exposed to the highest Ce concentration showed a clear loss of turgor pressure on the fifth day. An increase of hydrogen peroxide and malondialdehyde levels were observed in the plant shoots at 1200 µg/L of Ce. The highest concentration also triggered an answer by the shoots' antioxidant enzymes with a decrease in the activity of superoxide dismutase and an increase in peroxidase. However, no significant change in catalase activity was observed, compared to the control group, which may indicate that peroxidase played a more crucial role against the oxidative stress than catalase. Combined results indicate that H. portulacoides was actively responding to a toxic effect imposed by this higher Ce concentration. Nevertheless, changes in normal environmental conditions, may increase the bioavailability of Ce, while in areas where acid mine drainage may occur, the highest Ce concentration tested in this study may be largely exceeded, placing the sustainability of halophytes and estuarine marshes at risk.

Tissue and gender-related differences in the elemental composition of juvenile ocean sunfish (Mola spp.).

Trace elements are potentially critical contaminants of aquatic environments and fish, occupying upper trophic levels, are especially vulnerable to bioaccumulation. Due to public health concerns, however, data on the elemental composition of non-commercially important marine species are particularly lacking. Ocean sunfish (Mola spp.) attain a low commercial value worldwide and information on their elemental composition is limited. In this context, we examined the concentration of 11 trace elements (V, Cr, Mn, Co, Ni, Cu, Zn, As, Se, Cd and Pb) in eight tissues [brain, gills, gelatin (subcutaneous white gelatinous layer), gonads, spleen, liver, white muscle and red muscle] of 20 juvenile specimens (37.5-85.5 cm TL). Gender-related differences were solely found in the gonads and chiefly for essential elements possibly as a result of their importance in embryo development. Overall, Zn and As were the elements observed in greatest concentrations in body tissues. The considerably high presence of As should be related to the dietary preferences of juvenile ocean sunfish. Considerable inter-individual variability in the concentration of each element in any given tissue was observed, especially in the liver, likely originating from the inclusion of both benthic and pelagic prey in the diet of analysed fish. Greatest elemental loads were found in the liver and gills whereas lowest loads were observed in white muscle, brain and gelatin. Moreover, a clear distinction in elemental load and elemental composition was observed between white and red muscles, likely deriving from existing divergent metabolism-related physiological adaptations linked to their different roles in locomotion.