Unravelling the drivers of variability in body condition and reproduction of the European sardine along the Atlantic-Mediterranean transition.

Body condition and reproduction data are broadly used to assess the health status of fish because of its implications for recruitment and ecosystem structure. Sardina pilchardus is a small pelagic distributed throughout both Mediterranean and Eastern Atlantic. Seasonal trend analysis of energy storage and reproduction was carried out in sardines from two areas along the Atlantic-Mediterranean transition: Southern Portugal-Gulf of Cádiz (POR-GC) (Atlantic Ocean) and Alboran Sea (Alb) (Mediterranean Sea) from 2019 to 2021. Energetic condition was estimated using tissue and mesenteric fat content, hepatosomatic index (HSI), and the relative condition factor (Kn). Sex, reproductive developmental stage, and gonadosomatic index (GSI) were also obtained. In addition, the oceanographic and meteorological characteristics of the areas were analysed. Results showed that seasonal Kn, tissue and mesenteric fat content, and HSI values of POR-GC specimens exceeded Alb's with summer arrival, period in which sardine acquires reserves to allocate them to reproduction. These differences could be associated to greater productivity of the former area mainly due to rivers discharges and trade winds intensification during summer (from July to September). Furthermore, gonad maturation of POR-GC stock occurred before the Alb. However, no spawning capable individuals were identified until February in POR-GC. In contrast, in Alb it was observed a remarkable fraction of spawning capable and active spawner individuals in October. We hypothesized the migration of mature individuals from POR-GC to the spawning areas located in the Alb. Seasonal genetic population studies are required to untangle it and reliably evaluate the environmental effect on the stocks.

Epigenetic differences in the innate response after immune stimulation during zebrafish sex differentiation.

Infections are able to trigger epigenetic modifications; however, epigenetic-mediating infections in the immune system in fish is currently unavailable. Within this purpose, zebrafish were immune-stimulated with three lipopolysaccharides (LPS) during sex differentiation. Methylation patterns of three immune genes were studied by a candidate gene approach together with gene expression analysis, and in adulthood, sex ratios were determined. It was shown that the entrance of LPS was through the gills and accumulated in the pronephros. Significant hypomethylation levels of CASP9 and a significant CpG site for IL1ß after Pseudomonas aeruginosa LPS exposure were found. No methylation difference was observed for TNFα. Gene expression and correlation data differed among studied genes. Sex ratios showed a feminization in dose and LPS strain-dependent manner. Here, it is provided epigenetic regulatory mechanisms derived by innate response and the first evidence of possible epigenetic interactions between the immune and reproductive systems.

Immune genes, IL1ß and Casp9, show sexual dimorphic methylation patterns in zebrafish gonads.

There is crosstalk between the immune and reproductive systems in which sexual dimorphism is a common pattern in vertebrates. In recent years, epigenetics has emerged as a way to study the molecular mechanisms involved in gonadal development, those responsible for integrating environmental information that contribute to assigning a specific sexual phenotype (either an ovary or a testis). The knowledge of epigenetic mechanisms in certain molecular processes allows the development of epigenetic markers. In fish gonads, the existence of reproduction-immune system interactions is known, although the epigenetic mechanisms involved are far from clear. Here, we used the zebrafish (Danio rerio) as a model to study the DNA methylation patterns in gonads of two well-known innate immune genes: IL1ß and Casp9. DNA methylation levels were studied by a candidate gene approach at single nucleotide resolution and gene expression analyses were also carried out. Results showed that there was clear sexual dimorphism in the DNA methylation levels of the two immune genes studied, being significantly higher in the testes when compared to the ovaries. In summary, and although further research is needed, this paper presents sexual dimorphic methylation patterns of two immune-related genes, thus sex-biased differences in methylation profiles should considered when analyzing immune responses in fish. Data showed here can help to develop epimarkers with forthcoming applications in livestock and fish farming production, for example, in immune fish diseases or sexual control programs as epigenetic molecular tools to predict environmental pressure in the gonads.