functionally characterize flagellins from distinct bacteria in order to improve vaccine efficiency in aquaculture. Thus, we compare the properties of flagellins from different origin; a non- pathogenic species (Marinobacter algicola) and two pathogenic ones (Sal- monella thyphimurium and Vibrio vulnificus), as well as two S. thypimurium flagellins with different deletions on its hipervariable domains. The results showed very different abilities on the activation of macrophages and granulocytes of the gilthead seabream (Sparus aurata). * Corresponding author. E-mail address: vmulero@um.es (V. Mulero) P-227. Effects of nodavirus (VNNV) infection on gene expression profile in the gilthead seabream cell line SAF-1 P. Morcillo, M.A. Esteban, A. Cuesta * . Fish Innate Immune System Group, Department of Cell Biology and Histology, Faculty of Biology, Regional Campus of International Excellence “Campus Mare Nostrum”, University of Murcia, 30100 Murcia, Spain Abstract Viral nervous necrosis virus (VNNV) is responsible for many economical losses in some important marine aquaculture species. The fish virus pro- duces the viral encephalopathy and retinopathy (VER) disease altering therefore the brain and retina structure and function. Further studies conducted in different models will help to understand the VNNV biology and the interaction with their hosts. Thus, in this work, we have investi- gated the effects of VNNV infection, and its analogous poly I: C (pI:C), on the gene expression profile of the gilthead seabream (Sparus aurata L.) cell line SAF-1. Cells were incubated with culture medium (control), 10 6 TCDI 50 VNNV/ml or 50mg/ml of pI:C for 24 h at 25  C. Total RNA was isolated and the gene expression measured by real-time PCR. Evaluated genes belonged to those involved in antiviral response (interferon (IFN), myxovirus resistance (Mx), interferon regulatory factor-1 (IRF-1), melanoma differentiation- associated protein 5 (MDA5), lipophosphoglycan 2 (LPG2) or mitochondrial antiviral signaling protein (MAVS)), inflammation, antigen presentation (MHC), Toll-like receptors (TLR), antimicrobial defense, oxidative stress, apoptosis or cellular adhesion. Overall, VNNV infection elicited a much higher gene up-regulation than pI:C. The most up-regulated genes by VNNV infection of SAF-1 cells were those related to the antiviral response, mainly Mx, IFN and LPG2. At a lower extent, among the immune-relevant genes, TLR9, MHC Ia or antimicrobial peptides (b-defensin, histones 1 and 2B) were also up-regulated by VNNV infection. Other genes related to cellular stress, apoptosis or cell-adhesion were slightly affected or even down-regulated. Our results demonstrate that further cellular processes, apart from the antiviral response, are activated in the SAF-1 cell line upon VNNV infection and deserve further characterization. Acknowledgements This work was co-funded by grants AGL2010-20801-C02-02 (co-funded by Ministerio de Economía y Competitividad and FEDER) and 04538/GERM/06 (Fundación Séneca). * Corresponding author. E-mail address: alcuesta@um.es (A. Cuesta) P-285. Effects of the antidepressant, fluoxetine, on immune parameters of the clam Venerupis philippinarum M. Munari, L. Ballarin, M. Marin, V. Matozzo * . Department of Biology, University of Padova, Padova, Italy Abstract Pharmaceuticals and personal care products (PPCPs) are a large group of substances used either by human for personal health and cosmetic reasons or by agribusiness to enhance growth or health of livestock. PPCPs are produced in large quantities and comprise numerous chemicals, including prescriptible drugs, veterinary drugs, diagnostic agents, fragrances, lotions, and cosmetics. Due to human activities, residues from both pharmaceu- tical manufacturing and hospitals, illicit drug use, veterinary drug use and agribusiness, large quantities of PPCPs enter aquatic environments. Among pharmaceuticals, fluoxetine (a selective serotonin reuptake inhibitor) is an antidepressant commonly used for the treatment of depression and other psychological disorders. Despite its wide use, there is lack of information about the effects of fluoxetine on bivalve immune parameters. In the present study, the effects of fluoxetine on some important immune pa- rameters of the clam Venerupis philippinarum were evaluated for the first time. Clams (25 per concentration) were exposed for 7 days to differing fluoxetine concentrations (0, 1, 5, 25, 125, 625 mg/L), and haemolymph was collected from the anterior adductor muscle. Eight pools of haemolymph (from 3 bivalves each) were prepared for each experimental condition, and total haemocyte count (THC), Neutral Red uptake (NRU), lysozyme activity in cell-free haemolymph (CFH) and haemocyte proliferation were measured. A significant increase in THC values was observed in clams exposed to 25 mg/L, with respect to controls. NRU decreased significantly in haemocytes of clams exposed to 1 and 5 mg/L, compared with controls, whereas NRU returned to control values in clams exposed to the highest fluoxetine concentrations. Haemocyte proliferation increased significantly in animals exposed to 25, 125 and 625 mg/L, with respect to controls. Conversely, no significant alterations were observed in CFH lysozyme ac- tivity. Although preliminary, the results obtained demonstrate that fluoxetine affected markedly immune parameters in clams, even at envi- ronmentally realistic concentrations. * Corresponding author. E-mail address: matozzo@bio.unipd.it (V. Matozzo) P-385. Effects of live and heat-inactivated lactic acid bacteria on turbot (Scophthalmus maximus L.) leucocytes E. Muñoz-Atienza 1, * , N. Lluch 2 , C. Araújo 1, 3 , P.E. Hernández 1 , C. Herranz 1 , L.M. Cintas 1 , S. Magadán 2, 4 . 1 Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040-Madrid, Spain; 2 Centro Oceanográfico de Vigo, Instituto Español de Oceanografía (IEO), 36390 Vigo, Pontevedra, Spain; 3 Universidade de Trás-os-Montes e Alto Douro, 5001-801 Vila Real, Portugal.; 4 Virologie et Immunologie Moléculaires, INRA, 78352 Jouy-en-Josas, France Abstract During the last years, the use of probiotics has been increased in the aquaculture field. They are considered good candidates to reduce the use of antibiotics and control fish diseases. Among the many benefits of pro- biotics, modulation of the immune response could be one of the most important points to be studied. In this respect, it is very important the development of in vitro assays that let us identify the most effective pro- biotic strain and reduce the economic and ethic costs of the in vivo assays. The purpose of this work was to determine, in vitro, the effect of eight heat-inactivated and viable Lactic Acid Bacteria (LAB) of aquatic origin on the innate immune parameters of a relevant fish species for Galician and European marine aquaculture. We evaluated the effects of LAB (belonging to the genera Enterococcus, Lactobacillus, Lactococcus, Leuconostoc, Ped- iococcus and Weissella) on the respiratory burst and phagocytic activities of turbot head-kidney leucocytes by nitroblue tetrazolium (NBT) reduction assay and flow cytometry, respectively. The incubation of turbot leucocytes with live and heat-inactivated LAB resulted in a significantly increase in the respiratory burst activity. Moreover, six out of eight viable LAB induced a higher activation of the respiratory burst in turbot leucocytes than heat- inactivated LAB. The exposure to viable LAB increased significantly the phagocytosis index (PI) compared to untreated cells and seven of them raised significantly the percentage of cells with phagocyted particles. With Abstracts / Fish & Shellfish Immunology 34 (2013) 1692–1752 1726