Modulación de la citotoxicidad natural de los leucocitos de dorada (sparus aurata L.) / Alberto Cuesta Peñafiel ; dirigida por José Meseguer Peñalber, Mª Angeles Esteban Abad.

Tesis-Universidad de Murcia.Consulte la tesis en: BCA. GENERAL. ARCHIVO UNIVERSITARIO. T.M. 2362

Effect of Light–Dark Cycle on Skin Mucosal Immune Activities of Gilthead Seabream (Sparus aurata) and European Sea Bass (Dicentrarchus labrax)

Changes in di erent immune activities in the skin mucus of gilthead seabream (Sparus aurata L.) and European sea bass (Dicentrarchus labrax L.) specimens exposed to a constant light–dark photoperiod (12 h L:12 h D) were studied. Samples were collected at 08:00 (light on), 14:00, 20:00 (light o ), 02:00, and again at 08:00 to determine immunoglobulin M (IgM) levels, several enzymes related to the immune system, and bactericidal activity. IgM levels were higher during the day in seabream and reached a minimum value at 20:00, but it was hardly a ected in sea bass. No significant variations were recorded in the levels of protease and antiprotease. Peroxidase reached its maximum level in seabream at 02:00, the same time that it reached its minimum level in sea bass. Lysozyme showed little variation in seabream, but it was significantly lower at 14:00 than during the rest of the cycle in sea bass. Finally, di erent interspecific variations on bactericidal activity against Vibrio harveyi were recorded. The findings demonstrate that the immune parameters present in skin mucus of these important fish species are a ected by the light–dark cycle and that there are substantial interspecies di erences

Molecular characterization of the T cell costimulatory receptors 1 CD28 and CTLA4 in the European sea bass

For the activation of T cells, it is necessary the specific recognition of the peptide by the T cell receptors (TCR) in the surface of antigen-presenting cells (APCs) and additional signals delivered by costimulatory receptors. In fish, knowledge about the presence of these costimulatory signals is limited and functional evidence almost absent. Thus, in this study, we have identified the stimulatory CD28 and the inhibitory cytotoxic T-lymphocyte-associated protein 4 (CTLA4) coreceptors in the European sea bass (Dicentrarchus labrax), and evaluated their transcription. In parallel, the transcription encoding for the T cell markers CD8 and CD4 was also evaluated. Both coreceptors showed the canonical architecture including a signal peptide, an immunoglobulin domain, a transmembrane region and a cytosolic tail. Protein predictions and phylogenetic tree identify them as true mammalian orthologues of CD28 and CTLA4. We found these genes constitutively expressed in all studied organs of European sea bass with high expression in lymphoid organs (thymus, spleen and head-kidney) and liver. The molecular expression pattern of these genes was up-regulated in head-kidney leucocytes stimulated with T mitogens as concanavalin A and phytohemagglutinin (PHA), but not with the B cell mitogen lipopolysaccharide (LPS). Fish challenged with nodavirus (NNV) evidenced a differential and opposing regulation of the cd28 and ctla4 transcription levels in the brain, the target organ for viral replication, and head-kidney. While cd28 transcription tends to decrease over the infection time in both organs the expression of the ctla4 gene tends to increase. Interestingly, the coreceptor expression is highly and significantly correlated to the transcription of the T cell markers. Our results highlight the important role of CD28 and CTLA4 as costimulatory receptors of T cells in European sea bass but further studies are deserved

Phagocytosis in teleosts. Implications of the new cells involved

Phagocytosis is the process by which cells engulf some solid particles to form internal vesicles known as phagosomes. Phagocytosis is in fact a specific form of endocytosis involving the vesicular interiorization of particles. Phagocytosis is essentially a defensive reaction against infection and invasion of the body by foreign substances and, in the immune system, phagocytosis is a major mechanism used to remove pathogens and/or cell debris. For these reasons, phagocytosis in vertebrates has been recognized as a critical component of the innate and adaptive immune responses to pathogens. Furthermore, more recent studies have revealed that phagocytosis is also crucial for tissue homeostasis and remodeling. Professional phagocytes in teleosts are monocyte/macrophages, granulocytes and dendritic cells. Nevertheless, in recent years phagocytic properties have also been attributed to teleost lymphocytes and thrombocytes. The possible implications of such cells on this important biological process, new factors affecting phagocytosis, evasion of phagocytosis or new forms of phagocytosis will be considered and discussed

Sub-lethal doses of polybrominated diphenyl ethers affect some biomarkers involved in energy balance and cell cycle, via oxidative stress in the marine fish cell line SAF-1

Polybrominated diphenyl ethers (PBDEs) are a class of persistent contaminants which are found all over the world in the marine environment. Sparus aurata fibroblast cell line (SAF-1) was exposed to increasing concentrations of PBDEs 47 and 99, until 72 hours to evaluate the cytotoxicity, reactive oxygen species (ROS) production and the expression of some selected molecular markers related to cell cycle, cell signaling, energetic balance and oxidative stress (p53, erk-1, hif-1α and nrf-2), by real-time PCR. Furthermore, SAF-1 cells were exposed for 7 and 15 days to sub-lethal concentrations, in order to evaluate the response of some biomarkers by immunoblotting (p53,ERK-1, AMPK, HIF-1α and NRF-2). After 48 and 72 hours, the cells showed a significant decrease of cell vitality as well as an increase of intracellular ROS production. Gene expression analysis showed that sub-lethal concentrations of BDE-99 and 47, after 72 hours, up-regulated cell cycle and oxidative stress biomarkers, although exposure to 100μmol L-1 down-regulated the selected markers related to cell cycle, cell signaling, energetic balance. After 7 and 15 days of sub-lethal doses exposure, all the analyzed markers resulted affected by the contaminants. Our results suggest that PBDEs influence the cells homeostasis first of all via oxidative stress, reducing the cell response and defense capacity and affecting its energetic levels. This situation of stress and energy imbalance could represents a condition that, modifying some of the analyzed biochemical pathways, would predispose to cellular transformation

The short-term effects of farmed fish food consumed by wild fish congregating outside the farms

We simulated in the laboratory the possible effects on fatty acids and immune status of wild fish arriving for the first time in the vicinity of a sea-cage fish farm, shifting their natural diet to commercial feed consumption, rich in fatty acids of vegetable origin. The flesh fatty acid profile of golden mullet specimens was altered after 2 weeks of commercial feed consumption, showing an increase in fatty acids of vegetable origin. The serum peroxidase and bactericidal activities, and head-kidney leucocyte phagocytic capacity, increased after eight weeks of the new diet, while the respiratory burst activity decreased. The extent of these changes cannot be considered large enough to regard them as compromising the health status of fish. More research is needed in order to elucidate whether the rapid assimilation of the dietary fatty acids could harm the immune status of fish when feeding for longer periods than two months

Evaluación de la influencia de diferentes condiciones de cultivo en el potencial postbiótico de productos extracelulares de Shewanella putrefaciens Pdp11

Shewanella putrefaciens Pdp11 es una cepa probiótica que ejerce importantes efectos beneficiosos en peces cultivados. Sin embargo, no hay conocimiento sobre su potencial postbiótico en la industria acuícola. Dado que la producción de metabolitos postbióticos puede verse afectada por diferentes factores, el presente trabajo estudia los efectos en diferentes condiciones de crecimiento: (i) medios de cultivo, (ii) temperatura, (iii) tiempo de incubación y (iv) salinidad sobre el potencial postbiótico de los productos extracelulares (EPCs) de Pdp11. Para ello, se han analizado sus capacidades hidrolíticas, antibacterianas, antivirales y citotóxicas. Los resultados obtenidos han demostrado que las potenciales características postbióticas de Pdp11 dependen de las condiciones de crecimiento estudiadas, mostrando diferentes perfiles de hidrólisis enzimática, un efecto antivírico en el título del Virus de la Necrosis Nerviosa (NNV), un efecto estimulante en la viabilidad de diferentes líneas celulares, y un efecto en la formación de biopelícula de patógenos de peces.Este trabajo ha sido financiado por MINECO (#PID2020-113637RB-C22). Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tec