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The T-box transcription factor T-bet is expressed in a number of hematopoietic cell types in mammals and plays an essential role in the lineage determination of Th1 T-helper cells and is considered as an essential feature for both innate and adaptive immune responses in higher vertebrates. In the present study, we have identified and characterized the full-length Atlantic salmon T-bet cDNA (3502 bp). The putative primary structure of the polypeptide deduced from the cDNA sequence contained 612 aa, which possessed a T-box DNA binding domain. Phylogenetic study and gene synteny revealed it is as a homolog to mammalian T-bet. Quantitative PCR analysis of different tissues in healthy fish showed that salmon T-bet gene was highly expressed in spleen, followed by head kidney, and was expressed in intestine, skin, and liver at lower levels. Moreover, the time-dependent expression profile of T-bet, interferon gamma (IFNγ), interleukin-22 (IL-22), and natural killer enhancement factor in mucosal tissues during water-borne infection with live Aeromonas salmonicida, indicated the involvement of T-bet in mucosal immune response in Atlantic salmon.

Feed intake, rumen function, microbial protein (MCP) production and the efficiency of MCP production were determined in steers fed four different forage hays varying markedly in crude protein content. Low quality tropical forage (speargrass and Mitchell grass) hays had lower crude protein content, higher neutral detergent fibre content and lower digestibility than a medium quality tropical forage (pangola grass) hay and a temperate forage (ryegrass) hay. Steers fed speargrass and Mitchell grass hays had lower MCP production (80 and 170 g MCP/day, respectively) and efficiency of MCP production [78 and 79 g MCP/kg digestible organic matter (DOM), respectively] than steers fed pangola grass (328 g MCP/day; 102 g MCP/kg DOM) and ryegrass (627 g MCP/day; 135 g MCP/kg DOM) hays, which was directly related to the supply of DOM and rumen degradable protein. Intake was greatest for ryegrass hay, followed by pangola grass, Mitchell grass and speargrass hays [17.6, 15.6, 10.1 and 5.5 g DM/kg W.day, respectively]. The retention time of DM in the rumen was 72.1, 47.7, 28.6 and 19.1 h for speargrass, Mitchell grass, pangola grass and ryegrass hays, respectively, with a similar trend apparent for the retention time of neutral detergent fibre, lignin, chromium-EDTA and ytterbium labelled digesta. The difference in the protein : energy ratio of absorbed substrates (measured as efficiency of MCP production) did not appear to account for all the differences in intake, nor did a purely physical mechanism.

A laboratory ring trial was performed in four laboratories for determination of Cry1Ab toxin in leaf material of MON 810 maize using a standardised enzymelinked immunoassay protocol. Statistical analysis was carried out by the ISO 5725-2 guidelines, sample standard deviation and standard error, withinlaboratory and inter-laboratory SD and SE were calculated. Measured interlaboratory average values were 12.594.0, 15.394.6 and 72.6917.8 mg/g for three lyophilised samples, and 27.894.3 mg/g for a frozen sample, yet, Cry1Ab concentrations ranged 66.5Á160.1% of the corresponding IA. Determined concentrations by in-house protocols were statistically not different in one laboratory and different in two laboratories from the corresponding values by the joint protocol. Results emphasise the importance of a standardised protocol among laboratories for comparable quantitative Cry1Ab toxin determination. However, even when using a standardised protocol, significant differences still occur among toxin concentrations detected in different laboratories, although with a smaller range of variation.

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The overexpression of GATA-3, T-bet and TGF-ß may theoretically induce IL-4/A, IFN-γ and IL-17A expression, respectively. Whether this also applies to fish is not yet known. The plasmid vectors encoding reporter gene (RFP)-tagged T-bet, GATA-3 and TGF-ß were used as overexpression tools, transfected into cells or injected intramuscularly to monitor the expression of IFN-γ, IL-4/13A and IL-17A. In addition, the fish were either experimentally challenged with Vibrio anguillarum (VA group) or Piscirickettsia salmonis (PS group). The reporter gene (RFP) inserted upstream of the GATA-3, T-bet and TGF-ß genes, was observed in muscle cell nuclei and in inflammatory cells after intramuscular (i.m.) injection. PS group: following the injection of GATA-3 and T-bet-encoding plasmids, the expression of GATA-3 and T-bet was high at the injection site. The spleen expression of IFN-γ, following the injection of a T-bet-encoding plasmid, was significantly higher on day 2. VA group: The T-bet and GATA-3-overexpressing fish expressed high T-bet and GATA-3 mRNA levels in the muscles and on day 4 post-challenge. The expression of TGF-ß in the muscles of fish injected with TGF-ß-encoding plasmids was significantly higher on days 7 (8 days pre-challenge) and 19 (4 days after challenge). The protective effects of the overexpression of T-bet, GATA-3 and TGF-ß on both bacterial infections were negligible.