ach primer and 5 ul of cDNA. The PCR program consisted of an initial DNA denaturation of 94 C for 90 s, fol lowed by 45 cycles at 95 C for 30 s and 60 C for 60 s. A triplicate of the amplification reaction was realised for each sample. Plasma lysozyme concentration done Plasma lysozyme activity was determined at ambient temperature using a turbidimetric assay, adapted to microtitration plates. Briefly, a bacterial suspension of Micrococcus lysodeikticus was prepared at a concentration of 1. 25 g. l 1 in a 0. 05 M sodium phos phate buffer, pH 6. 2. Fifty microlitres of the samples were placed in 96 well microtitration plates. The reac tion was initiated in a Labsystems iEMS analyser, by addition of 160 ul well 1M. Lysodeikticus suspension using an automatic dispenser.

The optic density reading was taken at a wavelength of Inhibitors,Modulators,Libraries 450 nm every 15 s for 3 min, the plate being shaken before each reading. Lysozyme values for samples were converted to mg. ml 1, using a reference Inhibitors,Modulators,Libraries curve established with hen egg white lysozyme. Plasma alternative complement pathway activity Determination of the alternative pathway of plasma complement activity was carried out at 4 C through a haemolytic assay with rabbitred blood cells as described by Yano, adapted to microtitration plates. Sea bass samples, diluted to 1 64 in EGTA Mg GVB buffer to avoid nat ural haemolytic activity, were added in increasing amounts, from 10 to 100 ul well 1. Wells were filled with EGTA Mg GVB buffer to a final volume of 100 ul. Finally, 50 ul of 2% RRC suspension was added to each of the wells.

Control values of 0% and 100% haemolysis were obtained using 100 ul of EGTA Mg GVB buffer and 100 ul of non decomplemented trout haemolytic serum at 1 50 Inhibitors,Modulators,Libraries in ultrapure water, respectively. The samples were then incubated for 1 h at 20 C. The microplates were centrifuged and 75 ul of supernatant from each well was then Inhibitors,Modulators,Libraries transferred into another 96 well microplate with 75 ul of phosphate buffered saline. The absorbance was read in a Labsystems iEMS analy ser and the number of ACH50 units per ml of plasma was determined by reference to the 50% haemolysis. Dicentrarchus labrax oligonucleotide microarray Gene expression profiles were investigated using the Agilent 019810 Dicentrarchus labrax oligo microarray. This platform represents 19, 035 unique transcripts of the European sea bass.

Two non overlapping probes were designed for each tran script for a total 38, 070 oligonucleotide probes synthesized onto the array. All sequences are publicly available in a dedi cated database, Anacetrapib together with associated annota inhibitor purchase tions, GO entries and putative homologous genes in fish model species. Microarrays were synthesized in situ using Agilent non contact ink jet technology with a 4 �� 44 K format, and included default positive and negative controls. Microarray analysis was based on a single color design. A mixture of 10 different viral polyadenilated RNAs was also added to each RNA sample to monitor labelling and hybridization qual