Effect of dietary fish meal and fish oil replacement on lipogenic and lipoprotein lipase activities and plasma insulin in gilthead sea bream (Sparus aurata)

The effects of a double replacement of fish oil (FO) and fish meal (FM) by dietary vegetable ingredients in juvenile gilthead sea bream (Sparus aurata L. 1758) on some indices of lipid metabolism and plasma insulin levels were analysed. Four experimental diets with a replacement of 75% of FM by plant proteins (PP) were administered. Added oil was either FO (75PP/FO diet), or a vegetable oil mix (VO), replacing 33%, 66% or 100% of FO (75PP/33VO, 75PP/66VO, 75PP/100VO diets). Another diet with 50% of substitution of FM by PP and with 100% of VO was also tested (50PP/100VO diet). Final body weight was similar in all diet groups, except for the 75PP/100VO group, which presented lower values. Circulating insulin levels increased with feed administration in all groups and no differences between diets were observed, with the exception of the 75PP/FO group, which presented higher plasma insulin values. In adipose tissue, glucose‐6‐phosphate dehydrogenase and malic enzyme activities decreased with the inclusion of vegetable oil, especially 5 h after feeding. Diet had no significant effect on the hepatic activity of either enzyme. Lipoprotein lipase activity decreased in white muscle and adipose tissue with the replacement of fish oil in 75PP diets, 5 h after feeding. In conclusion, the use of a combined replacement of fish oil and fish meal by vegetable ingredients in gilthead sea bream permits satisfactory growth, with moderate changes in tissue lipogenesis and lipid uptake.     

Effect of masking organoleptic properties of fat on diet self‐selection by the sparid Diplodus puntazzo

Fish are able to select a balanced diet according to their nutritional needs by choosing among incomplete feeds or even pure macronutrients. However, the relevance of both the organoleptic properties of diet and the postingestive signals that they produce remains unclear. Thus, sharpsnout seabream were allowed to select between diets containing different edible oils with their organoleptic properties masked by using gelatine capsules. Fish were fed capsules of two different colours so that they could associate the capsule colour with its corresponding postingestive effect. The longitudinal experiment included a first phase during which the fish were adapted to consuming the gelatine capsules. In a second phase, the fish were challenged with two different encapsulated diets: one comprising a complete diet containing fish oil and the other a fat‐free diet. Sharpsnout seabream showed a preference for the fish oil capsules (3.8 ± 1.1 g kg^?1 body weight (BW), 66.8% of total intake) over the fat‐free capsules, showing that they were able to associate the colour of the capsule with their nutritional content through postingestive signals. After that, the fish were challenged to select between the capsules containing the fish oil diet and capsules containing a vegetable oil (linseed or soybean), in which case they showed no preference between diets (2.4 ± 0.3: 2.1 ± 0.5 g kg^?1 BW of fish oil versus linseed oil capsules and 2.2 ± 0.2: 1.8 ± 0.6 g kg^?1 BW of fish oil versus soybean oil capsules), indicating that the fatty acid composition of the different oils was not sufficient to affect dietary selection through postingestive signals. So, in conclusion, when orosensorial information from food is absent, the fish are able to select between diets at a macronutrient level by using postingestive information. However, this information is not sufficient for distinguishing between diets that differ in the type of oil used.     

Physico‐chemical characterization and in vitro digestibility of commercial feeds used in weaning of marine fish

Six different commercial feeds used in weaning and postweaning of marine fish were analysed from the perspective of some of their main physico‐chemical features and in vitro digestibility. While the range of particle sizes in dry state closely agreed to those claimed by the producing companies, dramatic changes were observed after suspension in water. Sedimentation rates measured in some of the feeds with higher particle sizes may be too high to ensure intake by young fish. The label claims of the products overestimate the amount of total protein but underestimate that of lipids. The analysis of amino acid profiles evidenced important differences in total amino acid composition between feeds of the same company but a great similitude in the profile of free amino acids (FAA). The evaluation of leaching evidenced that the fraction of FAA, which represented <2% of dry matter of the feeds, is quickly released in water, acting mainly as attractants. The in vitro digestibility assay evidenced significant differences in the bioaccessibility of the nitrogen fraction in some of the feeds by the digestive proteases of the sea bream (Sparus aurata).     

Development of digestive biochemistry in the initial stages of three cultured Atherinopsids

The early development of the digestive biochemistry of three cultured Atherinopsids (two Odontesthes species and Chirostoma estor) was investigated. Relatively high lipolytic and high alkaline proteolytic activities were found in all species. In addition, gene expression of lipase and trypsin during the early development of C. estor not only indicates an early onset of the lipase activity but also demonstrates a major importance of protein digestion in this species. However, despite all of them share similar habitats, differences in their digestive activities were evident. The rise in the activities of brush border membrane enzymes correlated with the decrease in cytosolic activities, used as indicator of maturation of the digestive tract, was detected in both species of Odontesthes at 9 weeks after hatching. In contrast, no similar signs were measured in C. estor, this being in agreement to the long weaning period (nearly 5 months) reported for this species.     

Effects of dietary supplementation with probiotic live yeast Debaryomyces hansenii on the immune and antioxidant systems of leopard grouper Mycteroperca rosacea infected with Aeromonas hydrophila

Two trials were conducted to determine the efficacy of fish fed live yeast Debaryomyces hansenii strain CBS 8339 on immune and antioxidant systems in leopard grouper Mycteroperca rosacea infected with Aeromonas hydrophila. Juveniles (12±0.5 g) were fed with a control diet or a D. hansenii‐supplemented diet (10⁶ colony‐forming units per gram) for 5 weeks. The live weight of fish was registered on a weekly basis. After 4 weeks, fish from each treatment were immunocompromised with pathogenic A. hydrophila and further fed for 1 week in order to evaluate the effect on immunological and antioxidant parameters. Generally, the results showed enhanced growth performance in fish fed the diet containing yeast compared with the control. Addition of live yeast had no significant effect on the immunological parameters after 4 weeks of feeding. However, post infection with A. hydrophila fish fed the yeast‐supplemented diet resulted in a significant increase in the levels of plasmatic immunoglobulin M. Superoxide dismutase and catalase (CAT) activities were significantly higher in the yeast group. In this fish, CAT and heat shock protein 70 genes were up‐regulated before and after infection of A. hydrophila. The present study is the first one reporting that yeast (D. hansenii) can enhance immunity and resistance against A. hydrophila.     

A new record of the freshwater pearl mussel Margaritifera margaritifera L. (Bivalvia,Unionoida) from the River Narcea (Asturias,north‐western Spain)

1. A population of the freshwater pearl mussel Margaritifera margaritifera L. was found in the River Narcea in the province of Asturias (north‐western Spain). Primary analysis of the population shows that it is well distributed along the river and is particularly abundant in shady river banks with a gravel substrate. 2. The length of living mussels ranges between a minimum of 22 mm and a maximum of 116 mm, with most in the size class between 65 mm and 70 mm. There is a good proportion of young mussels, although no fertile females were found in the course of the investigation. 3. Although there is no specific legislation in Spain for the protection of M. margaritifera, it has recently been proposed to include M. margaritifera in the Spanish List of Threatened Species as ‘vulnerable’. 4. Although the River Narcea seems to provide suitable conditions for the pearl mussel (water quality, available salmonid hosts, etc.), the main conservation measures must be directed at avoiding river fragmentation from dams, channelization and organic enrichment from individual farms or towns situated close to the river bank. Copyright © 2000 John Wiley & Sons, Ltd.     

Nitrogen and phosphorous fertilizer improve pastures naturally growing under hazelnut trees

Abstract

This three-year study (2003–2005) aimed to improve the yield and quality of pastures growing naturally that are colonized by naturally occurring vegetation without agricultural input under hazelnut (Corylus sp.) orchards in the middle and eastern Black Sea regions of Turkey. There were eight treatments: 1) control; 2) fertilizer only (triple superphosphate 44% and calcium ammonium nitrate 26%) (80kg ha^?1 P and 60 kg ha^?1 N in Samsun; 100 kg ha^?1 P and 80 kg ha^?1 N in Ordu; 100 kg ha^?1 P and 40 kg ha^?1 N in Giresun); 3) lime only (calcium carbonate 94%) (3.0 t ha^?1 lime in Samsun; 4.5 t ha^?1 lime in Ordu and Giresun); 4) early cut only; 5) soil aeration only; 6) fertilizer+lime; 7) fertilizer+lime+early cut; 8) fertilizer+lime+soil aeration, laid out in a randomized complete block design with four replicates at each location. The highest dry matter (DM) yield of kg ha^?1 and crude protein content (%) was obtained from the treatments that included fertilizer. There was no difference in DM production between any of the combination treatments that involved fertilizer and the fertilizer alone treatment. Only lime and aeration applications also increased yield compared to control, but not as much as did any treatment including fertilizer. Crude protein content of the pasture ranged from 13.3 to 18.1% across locations. Nitrogen and phosphorus fertilizer are recommended to improve DM yields and herbage quality for pastures under hazelnut orchards.     

Characterization of single‐nucleotide polymorphism markers in the Mediterranean mussel,Mytilus galloprovincialis

The Mediterranean mussel, Mytilus galloprovincialis, is one of the most important aquaculture species in Europe. Appropriate molecular markers are required to evaluate genetic resources and to trace genealogies in breeding programmes for improving mussel culture. Microsatellites have been commonly applied to this purpose in other species. However, Mediterranean mussel microsatellites have demonstrated high frequencies of null alleles that hamper accurate estimates of population parameters and confident parentage inferences. As alternative markers, we have characterized in silico 25 potential single‐nucleotide polymorphism (SNP) markers in the Mediterranean mussel from expressed sequence tag (EST) public databases. The genotyping of SNPs was performed using a single‐base extension approach. Their polymorphism was evaluated in 47 individuals from an Atlantic population. Out of the 25 potential SNPs tested, 12 were technically feasible (producing a single amplicon) and polymorphic. All were biallelic and had an unbiased heterozygosity ranging from 0.160 to 0.504. One SNP was from a mitochondrial gene. The combined potential of nuclear SNPs for parentage assignment gave an exclusion probability of a false couple of parents of 0.9471. These markers will be useful for evaluating resources and tracing genealogies in genetic breeding programmes implemented to solve the main problems of mussel culture.     

Potential bioavailability of protein and lipids in feed ingredients for the three‐spot cichlid Amphilophus trimaculatus: An in vitro assessment

For the commercial culture of the three‐spot cichlid (Amphilophus trimaculatus), it is important to have available a set of protein and lipid ingredients suitable for feed manufacturing, which allow to optimize the culture at affordable costs. The in vitro digestibility was evaluated with pH‐stat technique, using stomach and intestine multi‐enzymatic extracts from A. trimaculatus juveniles. The digestion of protein ingredients was taken place both in acid and in alkaline conditions, quantifying the total of free amino acids at the end of the hydrolysis, whereas the digestion of lipid ingredients was taken place only in alkaline conditions. The digestive enzymes of this species showed a better affinity to raw materials of animal origin such as beef meal, fish meal and premium chicken meal, which presented better digestibility. The best lipid ingredient that could be used is the fish oil. However, it is possible to use ingredients from the plant sources, such as soy or cereals, and vegetable oils such as olive, corn or soy oil. It is necessary to complement this study in vitro with the digestibility analysis in vivo, in order to determine the inclusion percentages in feeds and costs and therefore to develop an efficient feed formulation for the aquaculture of A. trimaculatus.     

Assessment of dietary lipid sources in tropical gar,Atractosteus tropicus larvae: Growth parameters and intermediary lipogenic gene expression

Five experimental diets containing different lipid sources, fish oil (D1), soybean lecithin (D2), corn oil (D3), canola oil (D4) and olive oil (D5), were evaluated in Atractosteus tropicus larvae for the relative gene expression of the enzymes fatty acid synthase (fas), acetyl‐CoA carboxylase 1 (acc1) and carnitine palmitoyltransferase 1C (cpt1c), in addition to their effects on larval growth, survival and cannibalism during a 30‐day feeding trial. Higher growth and survival were obtained in treatments D1 and D2, and lower performance in diets D3, D4 and D5. The highest levels of expression of fas and acc1 occurred in larvae fed with D1, which contained high amounts of n‐3 long‐chain polyunsaturated fatty acids (LC‐PUFA), mainly DHA and EPA FA are regulators of lipogenesis. The higher cpt1c expression in plant‐based diets is attributed to the fact that these diets are rich in α‐linolenic acid (ALA) and low DHA, EPA and ARA levels that favour ß‐oxidation. In conclusion, the diets with fish oil (D1) and soybean lecithin (D2) were the best treatments for larval growth, survival and cannibalism and thus appear to meet both lipid and energy requirements of A. tropicus larvae, meanwhile the use of vegetable oils influences the expression of intermediary lipogenic genes.