Project description:Fish in aquaculture farms frequently face unfavarouble husbandry conditions and other unpredictable situations, which are sometimes part of routine procedures. However, managing stress originated from these situations is crucial to ensure the sustainability of the production. When fish is exposed to prolonged stress, and overload of the physiological systems can occur and the fish may no longer be able to adapt and restore homeostasis, and this can impair the animal performance, such as growth and immunity, and consequently fish welfare. In this study the genes and gene families responsible for the molecular stress response to different challenges in gilthead seabream was assessed. Gilthead seabream adults were exposed to overcrowding, net-handling and hypoxia, in separate trials, each against a control group. Overcrowding and net-handling trials lasted for a month and half (chronic stress) and hypoxia for 48h (acute stress). The liver was the chosen organ for this transcriptomics analysis as this plays a crucial role in stress adaptation. The characterization of stress adaptation mechanisms provides valuable knowledge for the future selective breeding of more resilient commercial species that can thrive under changing conditions and adapt well to life in captivity, while ensuring high welfare standards.
Project description:A gilthead sea bream (Sparus aurata) microarray platform was developed to identify brain gene expression profiles in response to environmental concentrations of human pharmaceuticals. Comparative analysis of gene expression profiles was conducted among brain of gilthead seabream exposed to Acetaminophen (APAP; analgesic), Carbamazepine (CBZ; anti-epileptic) and Atenolol (AT; β-blocker). All groups of samples were also compared with brain of control individuals.
Project description:This study describes the development and validation of the Aquagenomic Sparus aurata oligonucleotide-microarray (SAQ) based on the Agilent Technology system (eArray) to provide a platform for studies on the gene expression of gilthead seabream. The platform developed used all available public ESTs stored and annotated in the Aquagenomic Consortium seabream library (10K). In fish, lipopolysaccharide (LPS) gives a robust cytokine response that is stimulated by crude LPS preparations, some component of the LPS complex are responsible for this stimulation. Peptidoglycan (PGN) is a component of G-negative bacteria (found as a contaminant of crude LPS preparations) able to be recognized by macrophages inducing depth transcriptional modulations and a strong inflammatory response. For microarray analysis, head kidney macrophage cultures were used (N = 36 fish). Each cell culture was stimulated with equal concentrations of PGN and LPS from E. coli O111:B4 strain (10 ug/mL): non-stimulated cell cultures (control n = 9 fish), stimulated during 6 h with LPS (n = 9), and stimulated during 1 h (n = 9), and 6 h (n = 9) with PGN. A loop microarray design approach was used for the study. All experimental RNA samples were labelled with a single colour dye (Cy3) and each stimulated sample was compared to the control sample (pool without stimulation) labelled with the same dye (Cy3). Our microarray analyses identified differential transcriptional modulations in macrophages stimulated with both LPS and PGN at the level of differentially activated RNA transcripts related with the regulation of transcriptional program, prostaglandin synthesis or highlighting the expression of responsive gene-cassettes tightly related to LPS-PGN host recognition. A total of 43,398 oligonucleotide probes were used to construct a high-density seabream microarray based on the Agilent 4 × 44 K design format. Microarray hybridization validation was made by analyzing the gene expression profiles in primary cultures of seabream macrophages (MC). 7,285 transcripts with annotated sequences were spotted in triplicate onto the slide (total probes 21,855), as well as 8,377 ESTs without annotation, 183 enriched sequences (gene bank) with 15 replicated probes (total probes 2,745), and finally 1,417 internal control probes of Agilent (N = 43,398).
Project description:Deciphering the dietary immunomodulatory effects of a medicinal plant leaf extract (MPLE) obateined from sage (Salvia officinalis, Lamiaceae) and lemon verbena (Lippia citriodora, Verbenaceae) upon the gut-associated lymphoid tissue (GALT) of gilthead seabream (Sparus aurata).
