Feeding habits of the two-banded sea bream (Diplodus vulgaris) and the black sea bream (Spondyliosoma gantharus) (Sparidae) from the south-west coast of Portugal

The stomach contents of Diplodus vulgaris and Spondyliosoma cantharus were: analysed using three simple methods (numeric, gravimetric and frequency of occurrence) and a composite index (I.R.I - Index of Relative Importance). To compare the species, the Schoener index was used. The diet of D. vulgaris consisted mainly of ophiuroids, polychaetes, amphipods and echinoids, while polychaetes, amphipods and hydrozoans dominated in the case of S. cantharus. There were some size-related differences in S. cantharus feeding. Diet overlap was relatively slight, with significant differences in feeding between the two species, notably in terms of greater consumption of echinoderms by D. vulgaris and hydrozoans by S. cantharus. As is the case for the majority of sea breams, D. vulgaris and S. cantharus are characterised by a diverse diet in terms of prey reflecting available prey items in their environment.

Semi-pelagic longline selectivity for two demersal species from the Azores: the black spot sea bream (Pagellus bogaraveo) and the bluemouth rockfish (Helicolenus dactylopterus dactylopterus)

The selectivity of four hook sizes (STELL brand(1), Quality 2335, numbers 12, 9, 6 and 4) used in a semi-pelagic longline fishery was studied in the Azores. Two species were caught in sufficient numbers for modelling of selectivity: the black spot sea bream (Pagellus bogaraveo) and the bluemouth rockfish (Helicolenus dactylopterus dactylopterus). A maximum likelihood method was used to fit a versatile model which can be used to describe a wide range of selectivity curves; from bell-shaped to asymptotic. Significant differences in size selectivity between hooks were found for both species. In the case of Pagellus bogaraveo, the smallest hook (number 12) had the lowest catch rates and all hooks were characterised by logistic-type selectivity curves, with sizes at 50% selectivity of: 27.9, 30.4, and 32.8 cm for hooks numbers 12, 9 and 6, respectively. The number 9 hook was the most efficient for Helicolenus d. dactylopterus, with selectivity curves varying from strongly skewed to the right for the number 12 hook to logistic-type for the numbers 6 and 4 hooks. Sizes at 50% selectivity for this species were 16.8, 18.7, 20.7, and 22.0 cm. respectively. (C) 1999 Elsevier Science B.V. All rights reserved.

Species and size selectivity in a 'red' sea bream longline 'metier' in the Algarve (southern Portugal)

A longline 'metier' using small hooks for 'red' sea breams (Pagellus acarne and Pagellus erythrinus) in the Algarve (south of Portugal) was studied. Experimental longlining was carried out with three sizes of "Mustad" round bend, flatted, spade end Quality 2316 DT hooks (numbers 11, 13 and 15) and two types of bait: razor shell (Ensis siliqua) and mud shrimp (Upogebia pusilla). A total of 3 328 fish and at least 36 species were caught with 33 600 hooks fished in 28 longline sets. Five species of sea breams (Sparidae) accounted for 79% of the catch: Pagellus acarne, Pagellus erythrinus, Diplodus vulgaris, Spondyliosoma cantharus, and Boops boops. High catch rates of 20-30 fish per 100 hooks were made in a number of 1 200 hook longline sets, with total catch weights of 40 to more than 60 kg per set. In general, the smallest hook (number 15) had the highest catch rate. Bait type did not significantly affect the catch size distributions. Although more fish were caught with the razor shell bait, higher catch rates of 'red' sea breams were obtained with mud shrimp. Catch rates were also affected by the location of the fishing grounds and the time of the set, with the highest catch rates obtained when the longline was set within two hours before sunrise. A wide size range was caught for each species, with highly overlapped catch size frequency distributions for the three hook sizes used. Except for Spondyliosoma cantharus, few illegal-sized fish were caught, even with the smallest hook. The logistic model fitted by maximum likelihood was used to describe hook selectivity for Diplodus vulgaris, Pagellus acarne, Pagellus erythrinus, and Spondyliosoma cantharus. (C) Ifremer-Elsevier, Paris.

The reproductive biology of the two-banded sea bream (Diplodus vulgaris) from the southwest coast of Portugal

The study of the reproduction of Diplodus vulgaris (Geoff.) as part of a base-line study of the fishery resources of the southwest coast of Portugal, was based on the analysis of the spawning season, gonad maturation, size at maturity, fecundity and hermaphroditism. The spawning season is relatively long, from December to March, with peaks in January and February. No significant differences were found either in the sex ratio (M:F = 1.01) over the year or by size. The size at first maturity (L-50) for all sexes and undetermined individuals combined was 18.33 cm total length (TL), with no significant differences between males and females. The estimated L-50 is considerably greater than the minimum legal size in Portugal of 15.0 cm. Mean absolute fecundity (F-a) was 131 127 oocytes, ranging from 31 523 to 250 608. The relationship between absolute fecundity and total length (TL) (F-a = 25 398 TL-484 426) and somatic weight (SW) (F-a = 878.8SW-71 416) was of the linear type. The mean number of oocytes per gram of somatic weight was 526, ranging from 194 to 887. The reproductive strategy of this species is characterized by a rudimentary hermaphroditism with possible protandry, as evidenced by the existence of individuals in sexual transition.

First report of cleaning behaviour in white sea bream (Diplodus sargus)

Several studies have reported the existence of ectoparasites in the stomach contents of Diplodus sargus. The cleaning behaviour has, however, never been previously observed for this species. In this short study, we report the first observations of the cleaning behaviour of D. sargus. These observations were in two yachting marinas, located in the Portuguese western coast between the months of April and August. The cleaning behaviour was only observed towards two Mugilidae species, Chelon labrosus and Mugil cephalus.

Teleost fish larvae adapt to dietary arachidonic acid supply through modulation of the expression of lipid metabolism and stress response genes

Dietary fatty acid supply can affect stress response in fish during early development. Although knowledge on the mechanisms involved in fatty acid regulation of stress tolerance is scarce, it has often been hypothesised that eicosanoid profiles can influence cortisol production. Genomic cortisol actions are mediated by cytosolic receptors which may respond to cellular fatty acid signalling. An experiment was designed to test the effects of feeding gilthead sea-bream larvae with four microdiets, containing graded arachidonic acid (ARA) levels (0·4, 0·8, 1·5 and 3·0 %), on the expression of genes involved in stress response (steroidogenic acute regulatory protein, glucocorticoid receptor and phosphoenolpyruvate carboxykinase), lipid and, particularly, eicosanoid metabolism (hormone-sensitive lipase, PPARα, phospholipase A2, cyclo-oxygenase-2 and 5-lipoxygenase), as determined by real-time quantitative PCR. Fish fatty acid phenotypes reflected dietary fatty acid profiles. Growth performance, survival after acute stress and similar whole-body basal cortisol levels suggested that sea-bream larvae could tolerate a wide range of dietary ARA levels. Transcription of all genes analysed was significantly reduced at dietary ARA levels above 0·4 %. Nonetheless, despite practical suppression of phospholipase A2 transcription, higher leukotriene B4 levels were detected in larvae fed 3·0 % ARA, whereas a similar trend was observed regarding PGE2 production. The present study demonstrates that adaptation to a wide range of dietary ARA levels in gilthead sea-bream larvae involves the modulation of the expression of genes related to eicosanoid synthesis, lipid metabolism and stress response. The roles of ARA, other polyunsaturates and eicosanoids as signals in this process are discussed.

Hepatocyte nuclear factor 4alpha transactivates the mitochondrial alanine aminotransferase gene in the kidney of Sparus aurata

Alanine aminotransferase (ALT) plays an important role in amino acid metabolism and gluconeogenesis. The preference of carnivorous fish for protein amino acids instead of carbohydrates as a source of energy lead us to study the transcriptional regulation of the mitochondrial ALT (mALT) gene and to characterize the enzyme kinetics and modulation of mALT expression in the kidney of gilthead sea bream (Sparus aurata) under different nutritional and hormonal conditions. 5′-Deletion analysis of mALT promoter in transiently transfected HEK293 cells, site-directed mutagenesis and electrophoretic mobility shift assays allowed us to identify HNF4α as a new factor involved in the transcriptional regulation of mALT expression. Quantitative RT-PCR assays showed that starvation and the administration of streptozotocin (STZ) decreased HNF4α levels in the kidney of S. aurata, leading to the downregulation of mALT transcription. Analysis of the tissue distribution showed that kidney, liver, and intestine were the tissues with higher mALT and HNF4α expression. Kinetic analysis indicates that mALT enzyme is more efficient in catalyzing the conversion of L-alanine to pyruvate than the reverse reaction. From these results, we conclude that HNF4α transactivates the mALT promoter and that the low levels of mALT expression found in the kidney of starved and STZ-treated fish result from a decreased expression of HNF4α. Our findings suggest that the mALT isoenzyme plays a major role in oxidazing dietary amino acids, and points to ALT as a target for a biotechnological action to spare protein and optimize the use of dietary nutrients for fish culture.

Sterol regulatory element binding protein-1a transactivates 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase gene promoter

6-Phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFKFB)catalyzes the synthesis and degradation of fructose-2,6-bisphosphate, a key modulator of glycolysis-gluconeogenesis. To gain insight into the molecular mechanism behind hormonal and nutritional regulation of PFKFB expression, we have cloned and characterized the proximal promoter region of the liver isoform of PFKFB (PFKFB1) from gilthead sea bream (Sparus aurata). Transient transfection of HepG2 cells with deleted gene promoter constructs and electrophoretic mobility shift assays allowed us to identify a sterol regulatory element (SRE) to which SRE binding protein-1a (SREBP-1a)binds and transactivates PFKFB1 gene transcription. Mutating the SRE box abolished SREBP-1a binding and transactivation. The in vivo binding of SREBP-1a to the SRE box in the S. aurata PFKFB1 promoter was confirmed by chromatin immunoprecipitation assays. There is a great deal of evidence for a postprandial rise of PFKB1 mRNA levels in fish and rats. Consistently, starved-to-fed transition and treatment with glucose or insulin increased SREBP-1 immunodetectable levels, SREBP-1 association to PFKFB1 promoter, and PFKFB1 mRNA levels in the piscine liver. Our findings demonstrate involvement of SREBP-1a in the transcriptional activation of PFKFB1, and we conclude that SREBP-1a may exert a key role mediating postprandial activation of PFKFB1 transcription.

Sterol regulatory element binding protein-1a transactivates 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase gene promoter

6-Phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFKFB)catalyzes the synthesis and degradation of fructose-2,6-bisphosphate, a key modulator of glycolysis-gluconeogenesis. To gain insight into the molecular mechanism behind hormonal and nutritional regulation of PFKFB expression, we have cloned and characterized the proximal promoter region of the liver isoform of PFKFB (PFKFB1) from gilthead sea bream (Sparus aurata). Transient transfection of HepG2 cells with deleted gene promoter constructs and electrophoretic mobility shift assays allowed us to identify a sterol regulatory element (SRE) to which SRE binding protein-1a (SREBP-1a)binds and transactivates PFKFB1 gene transcription. Mutating the SRE box abolished SREBP-1a binding and transactivation. The in vivo binding of SREBP-1a to the SRE box in the S. aurata PFKFB1 promoter was confirmed by chromatin immunoprecipitation assays. There is a great deal of evidence for a postprandial rise of PFKB1 mRNA levels in fish and rats. Consistently, starved-to-fed transition and treatment with glucose or insulin increased SREBP-1 immunodetectable levels, SREBP-1 association to PFKFB1 promoter, and PFKFB1 mRNA levels in the piscine liver. Our findings demonstrate involvement of SREBP-1a in the transcriptional activation of PFKFB1, and we conclude that SREBP-1a may exert a key role mediating postprandial activation of PFKFB1 transcription.

Effect of diet composition and ration size on key enzyme activities of glycolysis-gluconeogenesis, pentose phosphate pathway and amino acid metabolism in liver of Sparus aurata

The effects of diet composition and ration size on the activities of key enzymes involved in intermediary metabolism were studied in the liver of gilthead sea bream (Sparus aurata). Highcarbohydrate, low-protein diets stimulated 6-phosphofructo 1-kinase (EC 2.7.1.11), pyruvate kinase (EC 2.7.1.40), glucose-6-phosphate dehydrogenase (EC 1.1.1.49) and 6-phosphogluconate dehydrogenase (EC 1.1.1.44) enzyme activities, while they decreased alanine aminotransferase (EC 2.6.1.2) activity. A high degree of correlation was found between food ration size and the activity of the enzymes 6-phosphofructo 1-kinase, pyruvate kinase, glucose-6-phosphate dehydrogenase (positive correlations) and fructose-1,6-bisphosphatase (EC 3.1.3.11) (negative correlation). These correlations matched well with the high correlation also found between ration size and growth rate in starved fish refed for 22 d. Limited feeding (5 g/kg body weight) for 22 d decreased the activities of the key enzymes for glycolysis and lipogenesis, and alanine aminotransferase activity. The findings presented here indicate a high level of metabolic adaptation to both diet type and ration size. In particular, adaptation of enzyme activities to the consumption of a diet with a high carbohydrate level suggests that a carnivorous fish like Sparus aurata can tolerate partial replacement of protein by carbohydrate in the commercial diets supplied in culture. The relationship between enzyme activities, ration size and fish growth indicates that the enzymes quickly respond to dietary manipulations of cultured fish.

Effect of diet composition and ration size on key enzyme activities of glycolysis-gluconeogenesis, pentose phosphate pathway and amino acid metabolism in liver of Sparus aurata

The effects of diet composition and ration size on the activities of key enzymes involved in intermediary metabolism were studied in the liver of gilthead sea bream (Sparus aurata). Highcarbohydrate, low-protein diets stimulated 6-phosphofructo 1-kinase (EC 2.7.1.11), pyruvate kinase (EC 2.7.1.40), glucose-6-phosphate dehydrogenase (EC 1.1.1.49) and 6-phosphogluconate dehydrogenase (EC 1.1.1.44) enzyme activities, while they decreased alanine aminotransferase (EC 2.6.1.2) activity. A high degree of correlation was found between food ration size and the activity of the enzymes 6-phosphofructo 1-kinase, pyruvate kinase, glucose-6-phosphate dehydrogenase (positive correlations) and fructose-1,6-bisphosphatase (EC 3.1.3.11) (negative correlation). These correlations matched well with the high correlation also found between ration size and growth rate in starved fish refed for 22 d. Limited feeding (5 g/kg body weight) for 22 d decreased the activities of the key enzymes for glycolysis and lipogenesis, and alanine aminotransferase activity. The findings presented here indicate a high level of metabolic adaptation to both diet type and ration size. In particular, adaptation of enzyme activities to the consumption of a diet with a high carbohydrate level suggests that a carnivorous fish like Sparus aurata can tolerate partial replacement of protein by carbohydrate in the commercial diets supplied in culture. The relationship between enzyme activities, ration size and fish growth indicates that the enzymes quickly respond to dietary manipulations of cultured fish.

The effects of stocking density in physiological parameters and growth of the endangered teleost species piabanha, Brycon insignis (Steindachner, 1877)

This study investigated the effects of stocking density on the growth and fatty acid (FA) of Brycon insignis metabolism. Fingerlings (360) were distributed into eight ponds at two stocking densities (105 and 210 g/m(3)). The analysis of growth showed that the condition factor (K) and the coefficient of variation (CV) for body mass were not affected by stocking density. However, final body mass and length, specific growth rate (SGR), and weight gain (WG) were higher in the low stocking density group, which also presented a higher feed efficiency (FE) and survival (S). By contrast, muscle protein levels were higher in the high stocking density group. The plasma and muscle lipid content were not affected by stocking density, but fish reared at lower stocking density presented higher lipid concentration in the liver, with no differences in hepatosomatic index values. Even with the differences observed in metabolic and growth parameters, plasma cortisol was not affected by stocking density. The FA profile in the muscle and liver neutral fraction were not affected by stocking density, but the FA in the polar fractions differed between the two stocking densities. In the liver, total polyunsaturated fatty acids (PUFA) and PUFA n - 3 increased in higher stocking density, mainly due to an increase in docosahexaenoic acid (DHA). In addition, PUFA n - 6 were also increased in the higher stocking density group, mainly due to an increase in arachidonic acid (AA) and docosadienoic acid (22:2n - 6). In the muscle polar fraction, the saturated fatty acids (SFA) and monounsaturated fatty acids (MUFA) decreased in the animals from the higher stocking density group, and this reduction was compensated by an increase in PUFA n - 3 and PUFA n - 6, mainly the FA with 20-22 carbons (20:4n - 6: 22:4:n - 6; 22:5n - 6, 22:5n - 3, and 22:6n - 3). A different profile was observed for the C18 PUFAs, mainly 18:2n - 6 and 18:4n - 6, which were higher in the lower density stocking group. The data suggest that when living in high stocking density, B. insignis differentially utilizes the hepatic lipids as energy source and remodels the membrane fatty acids, with higher amounts of DHA in the polar muscle fraction compensated for by a decrease in MUFA. The zootechnical and physiological indices reveal that the lower stocking density group achieve overall better performance. (C) 2010 Elsevier B.V. All rights reserved.

Indagine sull’influenza di differenti sistemi d’allevamento sui connotati nutrizionali, tecnologici, sensoriali e di freschezza dell’orata (Sparus aurata) allevata in Italia

On account of the commercial importance of gilthead sea bream (Sparus aurata) in Italy the aim of the present study is the evaluation of the quality of nutritional, technological, sensory and freshness aspects. Sea bream production is growing in the Mediterranean and the evaluation of its quality concerns both producers and consumers alike. The culture system greatly influences final product quality. In Italy most of sea bream culture is carried out in cages but there is also a production in land based facilities and in lagoons. In this study external appeareance differentiations are pronounced. Different results were found for nutritional aspects, for fatty acids and for mineral content. Some differences in the freshness indices are also found. Furthermore, organoleptic differences have been described between culture system.

Stato sanitario di Spigole (Dicentrarchus Labrax) ed Orate (Sparus Aurata) allevate in Grecia e in Italia in relazione alla presenza di agenti di Zoonosi ed Ectoparassiti patogeni

Le attività di ricerca della presente tesi di dottorato si sono focalizzata principalmente sulla parassitofauna dei pesci marini allevati in Grecia ed in Italia con particolare attenzione allo studio degli ectoparassiti di maggior rilievo sanitario in maricoltura ed alla ricerca di endoparassiti di potenziale interesse zoonosico, in particolare larve di nematodi Anisakidae del genere Anisakis. Nel corso del triennio sono stati sottoposti ad esami parassitologici 916 spigole (Dicentrarchus labrax) e 462 orate (Sparus aurata) prelevate presso diverse tipologie di allevamenti greci ed italiani. Per quanto concerne le spigole, la presenza di ectoparassiti è stata riscontrata nel 29,2% e nel 61,9% dei soggetti provenienti rispettivamente da impianti siti in Grecia ed in Italia, mentre le orate hanno presentato percentuali di positività rispettivamente del 87,5% e del 26,7%. Gli ectoparassiti dominanti sono risultati essere il monogeneo Diplectanum aequans nelle spigole ed il ciliato Cryptocaryon irritans e il monogeneo Furnestinia echeneis nelle orate, sebbene sia stato possibile studiare anche il coinvolgimento di altri ectoparassiti, quali il monogeneo Sparicotyle chrysophrii ed il dinoflagellato Amyloodinium ocellatum, nel determinismo di alcuni episodi morbosi. Le osservazioni istopatologiche hanno permesso di caratterizzare le lesioni causate dagli ectoparassiti a diverse intensità d’infestazione. Per quanto concerne la ricerca di parassiti zoonosici, con particolare riferimento agli stadi larvali di nematodi Anisakidae del genere Anisakis, si sono condotti esami parassitologici a livello di cavità viscerale e di muscolo laterale in tutti i soggetti provenienti da allevamenti in gabbia (626 soggetti, di cui 441 spigole e 185 orate). Tutti i soggetti esaminati sono risultati negativi, indicando come il rischio di infestazione da larve di nematodi anisakidi possa essere considerato trascurabile in spigole ed orate allevate in gabbia, come già dimostrato per il salmone atlantico (EFSA, 2010).