Osmoregulatory changes in wedge sole (Dicologoglossa cuneata Moreau, 1881) after acclimation to different environmental salinities

The osmoregulatory responses of 20 days of acclimation to environmental salinities of 5‰, 15‰, 25‰, 35‰ and 55‰ were assessed in juveniles of wedge sole ( Dicologoglossa cuneata Moreau, 1881). This sole shows a good capacity to adapt to this range of environmental salinities. A direct linear relationship between environmental salinity and plasma osmolality was observed, with a calculated isosmotic point of 10.4‰ (284 mOsm kg⁻¹). Na⁺, K⁺-ATPase activity in the gills followed a 'U-shaped' relationship with environmental salinity, and a direct linear relationship in kidney tissue. Plasma cortisol levels were elevated in fish held in extreme salinities, and glucose levels were higher only in the group maintained at the highest environmental salinity. In the liver, a decrease in glycogen, lactate and amino acid contents was observed in specimens acclimated to extreme salinities (5‰ and 55‰), suggesting mobilization of liver metabolites. Metabolite levels in white muscle showed a pattern similar to the liver, with lower values in specimens acclimated to extreme salinities. We conclude that wedge sole is strongly euryhaline, but acclimation to extreme salinities comes with an energetic cost.     

A highly accurate, single PCR reaction for parentage assignment in Senegal sole based on eight informative microsatellite loci

From a battery of microsatellite markers (100 loci), recently identified by our group, we have selected eight for parentage assignment in Senegal sole ( Solea senegalensis ). This tool is based on microsatellite loci obtained from four genomic DNA libraries and one cDNA library. Within the eight loci (six from anonymous genomic DNA sequences and two located in expressed sequence tags of known genes), we have found, in an analysis of a reproductive broodstock, between nine and 16 alleles. The expected heterozygosity was between 0.616 and 0.860. In addition, we have optimized the polymerase chain reaction (PCR) conditions to amplify all loci simultaneously in a single multiplex PCR reaction, and we have tested three lots of male and female (five to six individuals) and three offspring (50–60 larvae each). The use of the eight microsatellite loci, the possibility of amplifying them in a single PCR reaction and the high value of the exclusion probability (0.9992) make this multiplex PCR method a unique tool for parentage assignment.
Finally, analysing one meiotic gynogenetic progeny, we have determined the relative distance of six of these loci to the centromere, and we have also found that all of them are unlinked. All these characteristics confer this tool with a high accuracy for parentage studies and genetic population analyses of Senegal sole.     

Different levels of organic trace minerals in diets for Nile tilapia juveniles alter gut characteristics and body composition,but not growth

Thirty groups of twenty juveniles each of Nile tilapia (Oreochromis niloticus, initial average weight 23.93 ± 0.80 g) were fed diets containing increasing amounts (0%, 25%, 50%, 75%, and 100% of levels used by the tilapia feed industry) of organic trace mineral (Fe, Zn, Mn, Cu, Se) supplementation for 56 days. Growth was not affected by the dose of mineral supplementation. However, body protein increased in fish fed intermediate doses of organic minerals, reaching a maximum at 40.44% supplementation. Doses >50% showed mild damage to the intestinal tissue. Body Fe, Zn and Cu deposition were highest at 28.32%, 24.43% and 78.45% supplementation, respectively. Tilapia could modulate intestinal morphology when fed different doses of organic minerals. As the supplementation doses increased, hemosiderin staining increased in the distal intestine and total body Fe content, indicating that the distal region of the intestine plays an important role in iron excretion by tilapia. Supplementation of organic trace minerals at intermediate doses does not harm Nile tilapia growth, but excessive trace mineral supplementation can impair body protein retention and promote histological alterations in the gut.     

Effects of two diets on lipid composition and reproductive performance of brill (Scophthalmus rhombus) eggs

The effects of two diets with different lipid composition (squid Loligo gahi and one commercial pellet for breeders) on reproductive performance and egg lipid composition of brill (Scophthalmus rhombus) were compared. A total of 36 adult brill weighing 1482 ± 432 g were randomly distributed in the experimental tanks. A flow‐through system composed of four tanks with 4.2 m² of bottom area (5.0 m³ × 1.2 m water depth) was used. The experiment lasted for approximately 5 months (168 days). From the four mature females fed squid, three ovulated. Five females fed pellets matured and ovulated. Total ovulations were of 10 and 44, by the three females fed squid and the five ovulating female fed pellets respectively. Egg viability was higher, although not significantly, for females fed pellets. This indicates that pellets could be a better food for breeding females. In general, the egg total lipid content and the lipid classes composition did not seem to be affected by diet. In contrast, results indicate that not only diets but also individual females, regardless of the diet consumed, are determinant for egg fatty acid composition, clearly modulating their composition.     

Water-use efficiency in cork oak (Quercus suber) is modified by the interaction of water and light availabilities

We studied the interaction of light and water on water-use efficiency in cork oak (Quercus suber L.) seedlings. One-year-old cork oak seedlings were grown in pots in a factorial experiment with four light treatments (68, 50, 15 and 5% of full sunlight) and two irrigation regimes: well watered (WW) and moderate drought stress (WS). Leaf predawn water potential, which was measured at the end of each of two cycles, did not differ among the light treatments. Water-use efficiency, assessed by carbon isotope composition (delta(13)C), tended to increase with increasing irradiance. The trend was similar in the WW and WS treatments, though with lower delta(13)C in all light treatments in the WW irrigation regime. Specific leaf area increased with decreasing irradiance, and was inversely correlated with delta(13)C. Thus, changes in delta(13)C could be explained in part by light-induced modifications in leaf morphology. The relationship between stomatal conductance to water vapor and net photosynthesis on a leaf area basis confirmed that seedlings in higher irradiances maintained a higher rate of carbon uptake at a particular stomatal conductance, implying that shaded seedlings have a lower water-use efficiency that is unrelated to water availability.     

Reproductive performance and seasonal plasma sex steroid and metabolite levels in a captive wild broodstock of brill Scophthalmus rhombus L.

This study reports egg production by captive wild brill Scophthalmus rhombus, a potential new flatfish species for Southern Europe‐Mediterranean mariculture, as well as seasonal plasma levels of 17β–estradiol, testosterone, 11–ketotestosterone, proteins, triglycerides, glucose and lactate. A mean egg production of 102 800 eggs kg body weight⁻¹ was achieved during the 2005 spawning period (January–March), although a continuous egg supply could only be obtained from some females, which had a higher relative fecundity (261 019±10 393 eggs kg⁻¹) with 12–17 eggs batches released at a mean interval of 3.4 days. Most eggs were obtained with water temperatures ranging from 12 to 14°C, and under increasing temperatures (up to 2.9°C). Potential egg viability (70.1±2.9%), fertilization (72.2±3.4%) and hatching rates (31.9±3.9%) showed high variability, with potential viability tending to decrease as the water temperature increased (mainly between 16 and 17°C) and 0% hatching above 16.6°C. The endocrine changes that brill underwent during late gametogenesis, spawning and postspawning periods were similar to those reported in other Pleuronectiformes. This study establishes an important basis for further research on the biology and physiology of brill reproduction, directed towards the optimization of the breeding techniques used currently.     

Genetic variability for responses to short‐ and long‐term salt stress in vegetative sunflower plants

Sunflower (Helianthus annuus L.) has been rated as moderately salt‐resistant, and variability for salt resistance has been detected within this crop. However, variability in salt‐resistance mechanisms has not been assessed. Osmotic tolerance, the relation of salt resistance with whole‐plant Na⁺ and K⁺ distribution and tissue Na⁺ tolerance were investigated in several sunflower inbred lines. Plants were grown under controlled conditions, in pots with sand and perlite irrigated with salinized (NaCl, –0.65 MPa) nutrient solution. Osmotic tolerance was assessed from the initial effects of the salt treatment on plant elongation in eleven sunflower lines. Long‐term salinity responses were evaluated in four of those lines, by assessing whole‐plant growth. A principal components analysis (PCA) was run on relative‐to‐control growth data, and this information was used to establish a relative resistance ranking, which indicated lines HAR2 > HAR1 > HA64 > HAR5. Osmotic tolerance was observed in HA64 and HAR2. The lines showed different degrees of Na⁺ accumulation, it was very low in some of them, but relative salt resistance was not associated to this trait. Tissue Na⁺ tolerance was deduced by comparing the percentage of dead leaves as a function of leaf blade Na⁺ accumulation, and it was higher in HAR1 than in the rest. These results indicate that variability for salt‐resistance mechanisms exists in sunflower. Osmotic tolerance and tissue Na⁺ tolerance were detected in different lines, highlighting that such variability may be exploited for increasing salt resistance in this crop.     

Protein and amino acid composition from the mantle of juvenile O ctopus vulgaris exposed to prolonged starvation

Protein and amino acid composition of the mantle of juvenile O ctopus vulgaris (Cuvier, 1797) during fasting for 27 days were determined. Average protein content of octopus mantle was of 711.19 ± 46.80 g kg^?1 DW, and it decreased with increasing fasting days. The non‐essential amino acids content was higher (486.18 ± 11.08 g kg^?1 protein) than essential amino acids (425.82 ± 9.15 g kg^?1 protein) at the start of the experiment (unstarved animals). The results suggest that the amino acid profile of the mantle where the most abundant amino acids are Arg, His, Lys, Gly, Leu and Pro could indicate a prolonged fasting condition (>20 days) or poor nutrition of O . vulgaris. This study supports the idea of using mantle for metabolic needs of starved O . vulgaris suggesting that the degradation pathway of amino acids to pyruvate and tricarboxylic acid cycle intermediates was favoured contrary to the degradation pathway of ketogenic amino acids. Special considerations should be taken concerning Thr, Ile, Ser, Ala, Asx (Asp, Asn), Glx (Glu, Gln) (because of their fast intake) and Lys and His (due to their stable contents) during a prolonged period of fasting.     

Effect of dietary protein level and source on digestive proteolytic enzyme activity in juvenile Senegalese sole, Solea senegalensis Kaup 1850

The effect of dietary protein level and protein source on growth and proteolytic activity of juvenile Solea senegalensis was studied. In Experiment 1, fish were fed on four experimental diets containing increased protein levels (36, 46, 56 and 67%). In Experiment 2, Senegalese soles were fed on five diets with partial substitution of fish meal by soybean meal, soybean protein concentrate, soybean protein isolate, wheat gluten meal or pea protein concentrate. Results prove that growth and proteolytic activity in the distal intestine of fish were affected by the quantitative increase in dietary protein. The origin of protein source used in the elaboration of experimental diets affected both the amount and composition of the alkaline proteases secreted into the intestinal lumen; however, it did not decrease animal growth. Juvenile Senegalese sole showed capability to modulate digestive protease secretion when the concentration and/or source of dietary protein were modified. Quantity and quality of dietary protein affected protein hydrolysis in Senegalese sole intestine. This study establishes that 30% fish meal protein can be replaced by soybean derivatives without affecting intestinal proteases. Replacement with wheat gluten meal or pea protein concentrate should be taken cautiously, but further research is needed to establish whether growth performance and digestive enzyme physiology of Senegalese sole are affected by plant protein-supplemented diets in a long-term trial.