Patterns of gastric evacuation, digesta characteristics and pH changes along the gastrointestinal tract of gilthead sea bream (Sparus aurata L.) and European sea bass (Dicentrarchus labrax L.).

A comparative study of gastric evacuation rates (GERs) and digesta content, moisture and pH values along the gastrointestinal tract was performed between gilthead sea bream and European sea bass. In order to distinguish species-specific differences from diet-elicited effects, all parameters were determined under either a fishmeal diet or a carob seed germ meal diet that contained high levels of total and soluble non-starch polysaccharides. GERs were significantly different between species and they were not affected by diet. Similarly, species-specific patterns were revealed in the distribution of digesta and water content along the gastrointestinal tract. In sea bream, stomach digesta and water content decreased with time, whereas in sea bass stomach retained the highest digesta and water content throughout the sampling period. The anterior and distal intestine exhibited the lowest accommodating capacities of digesta and water in either species. Overall, sea bream performed stomach digestion at lower hydration levels and higher pH compared with sea bass. Diet affected stomach moisture in both species and pH of stomach digesta in sea bass and of all intestinal sections in sea bream. The results obtained indicated that water and inorganic ion exchanges through the gut may differentiate between the species and warrant further investigation.

Major histocompatibility complex variation at class II DQA locus in the brown hare (Lepus europaeus).

The major histocompatability complex (MHC) is a multigene family of receptors that bind and present antigenic peptides to T-cells. Genes of the MHC are characterized by an outstanding genetic polymorphism, which is considered to be maintained by positive selection. Sites involved in peptide binding form binding pockets (P) that are collectively termed the peptide-binding region (PBR). In this study, we examined the level of MHC genetic diversity within and among natural populations of brown hare (Lepus europaeus) from Europe and Anatolia choosing for analysis of the second exon of the DQA locus, one of the most polymorphic class II loci. We aimed at an integrated population genetic analysis of L. europeaus by (i) correlating MHC polymorphism to genetic variability and phylogenetic status estimated previously from maternally (mtDNA) and biparentally (allozymes, microsatellites) inherited loci; and (ii) comparing full-length exon amino acid polymorphism with functional polymorphism in the PBR and the binding pockets P1, P6 and P9. A substantial level of DQA exon 2 polymorphism was detected with two completely different set of alleles between the Anatolian and European populations. However, the phylogeny of full-length exon 2 Leeu-DQA alleles did not show a strong phylogeographic signal. The presence of balancing selection was supported by a statistically significant excess of nonsynonymous substitutions over synonymous in the PBR and a trans-species pattern of evolution detected after phylogenetic reconstruction. The differentiating patterns detected between genetic and functional polymorphism, i.e. the number and the distribution of pocket variants within and among populations, indicated a hierarchical action of selection pressures.

Mitochondrial DNA variation in Northeast Atlantic and Mediterranean populations of Norway lobster, Nephrops norvegicus.

Analysis of the genetic structure of the Norway lobster (Nephrops norvegicus), a marine crustacean with high commercial value, was undertaken to gain information regarding the differentiation of Atlantic from Mediterranean populations of marine invertebrates. Restriction fragment length polymorphism analysis of two mitochondrial DNA segments, 3.6 kilobases in total, was performed. Twelve populations from the North Sea, Irish Sea, Portuguese coast and Aegean Sea were analysed. Low levels of differentiation were found among them (F(ST) = 0.018, P < 0.001) and there were no signs of an Atlantic-Mediterranean divide or of an isolation-by-distance scheme of differentiation. Possible reasons for these low levels of differentiation can be found in the recent expansion of N. norvegicus populations. This is supported by the mismatch distribution of pairwise haplotype differences, as well as by the high mean haplotype diversity (h = 0.93) combined with medium nucleotide diversity (pi = 0.0057) (in comparison to values for marine crustaceans or teleosts) found in this study. This combination of high levels of haplotype diversity with moderate to low levels of nucleotide diversity has also been frequently attributed to a recent time of divergence for various marine species. No evidence was found for a Mediterranean refugium during glaciation periods, separate from the Atlantic, as has been reported for some marine species. The Irish Sea population was the most differentiated as a result of reduced levels of diversity. Results are also discussed in the light of future management of N. norvegicus stocks.

Molecular cloning and sequence of Sparus aurata skeletal myosin light chains expressed in white muscle: developmental expression and thyroid regulation.

Two full-length cDNA clones encoding the skeletal myosin light chain 2 (MLC2; 1452bp) and myosin light chain 3 (MLC3; 972bp) were isolated from a cDNA library prepared from gilthead sea bream Sparus aurata larvae. The MLC2 cDNA encoded a predicted protein of 170 residues that was 79% identical to rabbit MLC2 over the entire length and 87% identical within the Ca(2+)-binding region. The deduced amino acid sequence of MLC3 was 153 residues in length and was 91% and 69% identical to the zebrafish and rabbit MLC3, respectively. Northern blot analysis revealed that in adults both transcripts were expressed in fast white muscle only. MLC2 appeared earlier in development: MLC2 transcripts were detectable from the beginning of segmentation, whereas MLC3 transcripts did not appear until 27h post-fertilisation. At this developmental stage, a second MLC2 transcript of 0.89 kilobase-pairs was present. MLCs exhibited a different age-related pattern of response to varied thyroidal states, which were experimentally induced by the administration of 1 microg g(-1)body mass of thyroxine (T4) or triiodothyronine (T3), or 5 ng g(-1)body mass of the hypothyroidal compound thiourea; MLC3 expression was not significantly affected, whereas levels of MLC2 transcripts were significantly elevated in the white muscle only of juvenile sea bream after administration of T4. Although the mechanism of thyroidal regulation of MLC expression remains unknown, the present results suggest that different regulatory mechanisms exist for different MLCs.

Molecular cloning and sequence of gilthead sea bream (Sparus aurata) alpha-skeletal actin: tissue and developmental expression.

In the present study, the Sparus aurata alpha-skeletal actin was cloned from a mixed larvae complementary DNA library. The clone isolated was 1523 bp long with an open reading frame of 1134 bp coding for a 377-amino acid protein. The deduced amino acid sequence of sea bream alpha-actin is identical to Fugu alpha-actin-1. The expression of alpha-actin was initiated at the onset of segmentation. In adult fish, alpha-actin is expressed predominantly in white and red muscle.

RFLP analysis of mitochondrial DNA to evaluate genetic variation in striped red mullet (Mullus surmuletus L.) and red mullet (Mullus barbatus L.) populations.

The genetic differentiation of striped red mullet (Mullus surmuletus) and red mullet (Mullus barbatus) was investigated in 6 Mediterranean populations of each species by means of restriction fragment length polymorphism analysis of mitochondrial DNA. Three segments amplified by polymerase chain reaction (control region, COI, and 12S-16S ribosomal RNA) were digested with 20 restriction endonucleases, revealing 71 haplotypes for M. surmuletus and 30 for M. barbatus. For the two species nucleotide diversity was equally distributed within and among populations, leading to N(ST) values of 0.545 and 0.500 for M. surmuletus and M. barbatus, respectively. However, intrapopulation and interpopulation genetic structuring appeared to be much higher for M. surmuletus than for M. barbatus (1.88% vs. 0.46% of mean intrapopulation nucleotide diversity; 1.94% vs. 0.47% of mean interpopulation nucleotide diversity; 0.055% vs. 0.002% of net interpopulation divergence). Furthermore, 81.69% of the haplotypes observed for M. surmuletus were unique, whereas 70.29% of M. barbatus individuals were grouped in 3 common haplotypes. Given that fishing pressure and population sizes are similar for both species, this differentiation could be attributed to differences in biological parameters and life histories between the two species, coupled with oceanographic conditions prevailing in the studied area.