Effect of dietary canthaxanthin on the growth and lipid composition of red porgy (Pagrus pagrus)

A 24‐week feeding trial was conducted to study the possible effect of dietary canthaxanthin on red porgy growth and lipid composition. Two triplicate groups were established to test two experimental diets: (1) Control group fed a diet with no added carotenoids, and (2) canthaxanthin group (CTX100) fed a diet with 100 mg of synthetic canthaxanthin per kilogram of diet (CTX). Final and eviscerated weight were increased (P < 0.05) in the CTX100 treatment. The rest of growth performance parameters were not affected by the CTX diet. Whole‐fish total lipid content was decreased (P < 0.05) in CTX100 fish. In the liver, total lipids were not affected; however, saturated fatty acids in CTX100 treatment were significantly lower together with a higher n‐3 PUFA and a lower n‐6 PUFA, therefore increasing the n‐3/n‐6 ratio. Liver histology of CTX100 fish revealed decreased lipid vacuolization thus, significantly lowering hepatocyte area. In the muscle, total lipids were not affected. Similar to the liver, an increase of n‐3 PUFA and decrease n‐6 PUFA, led to a significant increase of the n‐3/n‐6 ratio. Concerning plasma, only total cholesterol (TC) was significantly affected by the CTX diet. Dietary canthaxanthin has an effect on red porgy lipid composition.     

First report of Streptococcus iniae in red porgy (Pagrus pagrus, L.)

This work describes the first isolation of Streptococcus iniae in red porgy, Pagrus pagrus (L.), and the first European isolation of this pathogen in gilthead seabream, Sparus aurata (L.). In both farmed fish species, infection resulted in lethargy, anorexia, abnormal swimming, exophthalmia and sudden death, with mortality rates of over 25% in red porgy and 10% in gilthead seabream. Beta-haemolytic Gram-positive cocci, catalase negative and oxidase negative, were isolated in pure culture from internal organs. Conventional and rapid identification systems, and 16S rRNA gene partial sequencing were used to identify the causative agent of the natural disease. LD50 trials were carried out to show the virulence of this isolated strain in these species, with values of 1.7 × 104 CFU per fish in red porgy and 1.32 × 105 CFU per fish in gilthead seabream. The most prominent lesions were meningoencephalitis and multifocal infiltration of macrophage cells in the kidney and spleen.     

Effects of different dietary protein and lipid levels on growth, feed utilization and body composition of red porgy (Pagrus pagrus) fingerlings

Two feeding trials were conducted to determine the minimum dietary protein level producing maximum growth, and the optimum protein to energy ratio in diets for red porgy (Pagrus pagrus) fingerlings, respectively. In the first trial, six isoenergetic diets were formulated with protein levels ranging from 400 to 650 g kg^?1 in increments of 50 g kg^?1, and fed for 11 weeks to 2.8 g average initial weight fish. Weight gain, specific growth rate and feed efficiency were significantly higher with diets containing higher protein levels, when compared with dietary levels below 500 g kg^?1. The highest protein efficiency ratios were obtained in fish fed 500 g kg^?1 dietary protein. The minimum dietary protein level producing maximum fish growth was found to be 500 g kg^?1. In the second trial, 15 g average initial weight fish were fed for 12 weeks, six diets containing three different lipid levels (100, 150 and 200 g kg^?1) combined with two protein levels (450 and 500 g kg^?1). Weight gain values increased when dietary lipids increased from 100 to 150 g kg^?1, with a further decrease for 200 g kg^?1 lipids in diets; the lowest fish growth being supported by 200 g kg^?1 dietary lipids. Fish growth was significantly higher when dietary protein increased from 450 to 500 g kg^?1. There was no evidence of a protein‐sparing effect of dietary lipids. Liver protein and lipid contents were low when compared with other fish species. All diet assayed produced high liver glycogen accumulation. The recommended protein and lipid levels in diets for red porgy fingerlings were 500 and 150 g kg^?1, respectively.     

Environmental control of skin colour in the red porgy, Pagrus pagrus

Fish display sophisticated skin chromatic properties that are of considerable ecological, physiological and behavioural importance. The aim of study was to investigate the role of important physical parameters (background colour, lighting spectrum, light intensity and water temperature) on skin colour to gain better knowledge of the environmental factors that regulate pigmentation in cultured red porgy, Pagrus pagrus . All tested physical parameters had a significant effect on skin lightness, especially in the dorsal body area; low light intensity, blue spectrum, a water temperature of 19 °C and a white background significantly increased skin lightness. This effect was mediated through changes in melanophore motility and/or skin melanin concentration. None of the examined factors affected skin hue while a significant decrease in skin entire colour index (a combination of hue and chroma) was found only in white background-adapted fish. It is concluded that all tested physical factors are important regulators of skin lightness.     

Influence of stocking density on growth of juvenile red porgy Pagrus pagrus L. under different feeding conditions

The influence of stocking density and feeding regime on the growth of red porgy was studied during winter and spring 1997. Fish were held in 500-L tanks at densities of either 50 or 100 fish per tank, and were fed by means of self-feeders under 6 h restricted (day or night) or free-access feeding regimes. Fish were tagged individually, and weight gain was recorded by individual weighing every second week over an 8-week period during each season. In winter, growth was influenced by feeding time. Fish tended to grow best when fed without restriction and worst when fed at night, although differences were not significant when tank means were compared. Stocking density was influential in the spring trial. Fish stocked at low density and feeding freely grew better than high-density fish that fed either freely or at night. The division of fish into three size classes revealed that, within each treatment, no differences occurred among classes, but growth of fish belonging to the same class was influenced by rearing conditions. No significant differences were found among feeding regimes in terms of the coefficients of weight variation ratio (CV_f:CV_i) and feed efficiency.     

Fatty acids in muscle of wild and farmed red porgy, Pagrus pagrus

The total fat content and the fatty acids of total lipids (TL), phospholipids (PL) and triacylglycerols (TAG) were analysed in white muscle from wild and reared red porgy, Pagrus pagrus (L.). The fat percentage was higher in reared (3.03 ± 0.57%) (mean ± SE) than wild (0.65 ± 0.03%) fish. The fatty acid pattern of reared porgy TL reflected the diet's lipid. Wild red porgy showed higher levels of ω3 and ω6 polyunsaturated fatty acids (PUFA) than reared, but both the atherogenic index and the index of thrombogenicity were very low for both wild and reared red porgy. The lipid fractions showed that the PL were composed of very high levels of PUFA, especially 22:6ω3, and their composition seemed to be better regulated than the composition of TAG. TAG showed greater differences between wild and reared fish, presumably because they reflected the dietary content of lipids. PUFA were the principal component of wild red porgy TAG.     

Separate and combined effects of cyclic fasting and l‐carnitine supplementation in red porgy (Pagrus pagrus,L. 1758)

We examined the effects of cyclic fasting in red porgy (Pagrus pagrus) fed different dietary carnitine levels. Juvenile fish (23.58 ± 3.49 g) were divided into eight groups – four groups were fed every day to apparent satiation, while the other four were fasted for 7 days every 2 weeks. In each feeding regime, two replicates were fed an l ‐carnitine non‐supplemented diet (46 mg kg^?1) and the other two groups were fed an l ‐carnitine supplemented diet (630 mg kg^?1). Fish fed 630 mg l ‐carnitine accumulated two times more l ‐carnitine in muscle than fish fed 46 mg l ‐carnitine. Cyclic fasting reduced the growth performance and lipid content in the liver. Carnitine supplementation did not affect performance and body composition, but decreased the n‐6 PUFA content. Moreover, the combined effects of fasting and carnitine supplementation were observed on reducing the n‐3 fatty acid content. Areas of steatosis were found in the livers of red porgy, but the results revealed that supplementation of l ‐carnitine in cyclic fasted fish contributed towards a lower degree of vacuolization than in fish fed to apparent satiation. Regardless of the feeding regime applied, the spleen of fish fed the l ‐carnitine‐supplemented diet was haemorrhagic and hyper activation of melanomacrophage cells was observed.     

Seasonal changes in sperm production and quality in the red porgy Pagrus pagrus (L.)

Cultured red porgy Pagrus pagrus (L.) males (n=6) were sampled every 2 weeks for milt, in order to monitor changes in sperm quality parameters during a whole spawning period. On 11 January 2001, 60% of the fish were spermiating, increasing to 100% in mid‐February and dropping to 30% by mid‐April. Sperm density showed a slight increasing trend, with mean values ranging between 8.6 and 23.7×10⁹ spermatozoa mL^?1. Sperm motility percentage exhibited a significant improvement during the spawning season (analysis of variance (anova ) P=0.0001). The duration of forward motility for the major part of the monitoring period ranged between 2 and 4 min. Red porgy spermatozoa maintained their viability for many days after whole storage of milt at 4°C. During the monitoring period there were significant changes in the mean duration of sperm survival after cold storage, ranging from 5 to 12 days. The total volume of expressible milt was maximal on 28 March, increasing from a mean value of 1.7 mL to 5.3 mL kg^?1. Milt production of captive‐reared red porgy does not appear to be limiting, when compared with the volume of expressible milt produced by other cultured marine fishes.     

Marine and freshwater crab meals in diets for red porgy (Pagrus pagrus): effect on growth,fish composition and skin colour

River crab (RC) meal (Procambarus clarkii) and marine crab (MC) meal (Chaceon affinis) were tested as a partial replacement for fish meal in diets for red porgy (Pagrus pagrus), and their effects on growth performance, fish proximate composition and skin colouration were evaluated. Red porgy were fed during 165 days with five diets. High‐quality fish meal diet was used as a control diet (CD). Protein of fish meal in the control was replaced by increasing the dietary levels of protein derived from RC and MC by up to 10% and 20% of each of them (RC10, RC20, MC10 and MC20). Fish fed on MC20 showed the highest values in feed intake, weight gain and growth (%). No differences were found in FCR and protein efficiency ratio among the treatments. Inclusion of both crab meals in diets significantly decreased the lipid content in whole fish compared with the control animals. On the other hand, no differences in muscle composition were found between the diets. Feeding both crab meals resulted in colour improvement compared with that of the control fish, with better hue values for the RC meal group than those for the MC meal group. The crab meals tested in the present study are suitable as a partial replacement for fish meal in diets for the red porgy, with the MC meal improving growth and both crabs meals improving skin colour, with further improvements in skin colour produced in fish‐fed diets containing the RC meal.     

Digestive enzyme activity of the red porgy (Pagrus pagrus, L.) during larval development under culture conditions

The ontogenic development of the main digestive enzymes (proteases, amylase and lipase) in the red porgy, Pagrus pagrus, larvae was assayed during the larval development. The green water technique was carried out for larval rearing and whole‐body homogenates were used for enzymatic assays in triplicate. Significant alterations in specific activities of all digestive enzymes measured during the period of this study were mostly related to metamorphosis and weaning. Trypsin‐ and chymotrypsin‐specific activities were first detected on day 3, together with opening of the mouth, and slightly increased until 25 days after hatching (DAH). After this period, the specific activities of these enzymes slightly decreased. Pepsin was first detected on day 28, concurrent with stomach formation, and a sharp increase was observed until 30 DAH. A slight decrease was measured from this date until the end of the experiment. Both amylase and lipase were measured for the first time on days 2 and 4 respectively, and the specific activities of these enzymes showed similar patterns during the first week of the study. Then, slight variations were observed until 30 DAH and while lipase‐specific activity declined, an increase in the specific activity of amylase was found until the end of the experiment. It is concluded that the variations observed in the specific activity of digestive enzymes were related to either metamorphosis, such as the formation of the stomach (28 DAH), or to changes in food composition. The profile of the developmental pattern of the main digestive enzymes detected in P. pagrus is similar to that described for other Sparid species.     

Influence of food deprivation and high stocking density on energetic metabolism and stress response in red porgy, Pagrus pagrus L

In the present study, the influence of high stocking density and food deprivation on red porgy (Pagrus pagrus L.) energy metabolism was investigated after 14 and 21?days assessing plasmatic, hepatic, and muscular parameters. Specimens were submitted to four different experimental conditions: (1) fed fish under low stocking density (4?kg?m^?3, LSD-F); (2) food-deprived fish under low stocking density (LSD-FD); (3) fed fish under high stocking density (50?kg?m^?3, HSD-F); and (4) food-deprived fish under high stocking density (HSD-FD). At plasmatic level, cortisol concentration enhanced in fish maintained under HSD condition, with higher effect in fed fish, whereas FD condition decreased values of lactate, protein, and triglycerides. In liver, significant increases in triglyceride levels, together with a decrease in glycogen values, were observed in FD specimens. Furthermore, a weak effect of density in fed fish was observed, with decreasing hepatic glycogen. In white muscle, aminoacid and triglyceride values diminished in FD group, while increased in specimens submitted to HSD condition. Our results suggested an enhancement in P. pagrus gluconeogenic and glycogenolytic potential when submitted to HSD and FD conditions to cope reallocation of metabolic energy due to these different stress situations.     

Physiological colour changes in the red porgy, Pagrus pagrus, following adaptation to blue lighting spectrum

The effect of lighting spectrum (full vs. blue) on skin colour and stress response in red porgy, Pagrus pagrus, held under a 12L:12D photoperiod was investigated. Fish exposed to blue lighting spectrum became gradually paler with a maximum lightness value (L = 45.33) on day 27, significantly higher than control fish (L = 31.49). However, there was no difference in skin melanin content, hue and chroma among the experimental groups. There was also no statistically significant effect of lighting spectrum on average glucose, thyroxine (T₄), triiodothyronine (T₃), cortisol (F) and Melanophore Stimulating Hormone (MSH) plasma concentrations nor in the pattern of changes during the time course of the experiment. Results showed that lighting spectrum affects skin lightness but not the chromaticity attributes hue and chroma. This effect is due to changes in the motility of the melanophores (the area occupied by melanosomes) and is not mediated by MSH or the general stress response.     

Pigmentation,carotenoids, lipid peroxides and lipid composition of red porgy (Pagrus pagrus) skin reared under open‐cage conditions

Pigmentation capability of red porgy (Pagrus pagrus) skin reared under open sea‐cage conditions and fed an astaxanthin‐enriched diet was studied. Skin lipid peroxide levels and lipid composition were also evaluated to establish the antioxidant role of astaxanthin under these sunlight‐exposure conditions. Fish placed either in an offshore sea cage system (SC) or in an inland tank facility (T) housed inside a ‘shade‐house’ enclosure were fed a commercial diet supplemented with 22 mg kg^?1 astaxanthin. No differences in growth or survival were found. Both groups displayed a red skin, but SC fish presented a darker pigmentation, which externally reflected the higher deposition of melanin, astaxanthin and tunaxanthin found in its skin. The lower level of lipid peroxides found in SC fish might be related with the higher level of astaxanthin mentioned above. Nevertheless, lipid and fatty acid profiles did not show significant differences between groups. Our results indicate that sustainable production of red porgy with a natural red hue is possible on the basis of proper adjustment of two factors illumination and dietary astaxanthin.     

The effect of different carotenoid sources on skin coloration of cultured red porgy (Pagrus pagrus)

This study presents data on the effect of carotenoid sources on skin coloration of red porgy (Pagrus pagrus). Three experiments were conducted: in the first, fish were fed an astaxanthin (Naturose^®)‐supplemented diet, while the second fish received diets supplemented with β‐carotene (Rovimix β‐caroten^®) or lycopene (Lyc‐O‐Mato^®): Carotenoids were added to the level of 100 ppm in each diet, while a non‐carotenoid‐supplemented diet served as a control. In the third experiment, the effect of dietary protein/carbohydrate ratio on melanin content in the skin was investigated. For this experimentation, four diets were formulated to contain 50/23, 40/32, 30/48 and 20/59 protein/carbohydrate ratio. Naturose^® astaxanthin increased total carotenoid content in the dorsal skin area while β‐carotene and lycopene seem to have had no significant effect. Naturose^® was the only carotenoid source that had a significant effect on skin hue, promoting a reddish coloration to the dorsal skin area and a ventral hue similar to wild red porgy. No apparent effect of carotenoid source on skin melanin content was observed. In contrast, dietary protein/carbohydrate ratio affected melanin content in the skin. The fish fed the 50/23 diet showed significantly higher values. Farmed red porgy had eight times higher dorsal‐skin melanin content than wild ones.     

Dietary carotenoids and skin melanin content influence the coloration of farmed red porgy (Pagrus pagrus)

The present study investigates the effects of dietary carotenoid sources on the coloration of the red porgy, Pagrus pagrus. Red porgies (131.9 ± 16.2 g; mean ± SD) were fed for 12 weeks on five different diets supplemented with red carotenoids (mainly astaxanthin esters) supplied from Haematococcus pluvialis algae and yellow carotenoids (mainly β‐carotene, lutein, and zeaxanthin) supplied from Alfalfa, Medicago sativa L. and Spirulina, Spirulina pacifica. The carotenoid‐supplemented diets did not have any marked effect on the growth rate, the feed conversion ratio, the daily feeding rate, or the hepatosomatic index of red porgy. The biochemical indices measured in plasma including cholesterol, total proteins, glucose, lactate, phospholipids, non‐esterified fatty acids, triglycerides, and thyroid hormones did not differ significantly among groups. Diet did not affect significantly the melanophore‐area coverage, the melanin skin concentration and skin lightness. Carotenoid‐supplemented diets affected significantly the carotenoid deposition in the skin, the presence and distribution of erythorphores and xanthophores, and skin hue and chroma. Overall, data have shown the efficacy of Haematococcus algae in promoting a reddish coloration in red porgy.     

Endocrine changes during the annual reproductive cycle of the red porgy, Pagrus pagrus (Teleostei, Sparidae)

Changes in 17-estradiol (E2), estrone (E1), testosterone (T), 11-ketotestosterone (11KT), and 17,20-dihydroxy-4-pregnen-3-one (17,20-P) levels were correlated to changes in gonadosomatic index (GSI), vitellogenin concentration (Vg), ovarian and testicular histology during the annual reproductive cycle of the red porgy, Pagrus pagrus. The production of E2, E1, T and 17,20-P was confirmed by analysis of the steroidogenic activity of ovaries. In females, the average concentration of E2 was lower than 2 ng ml^–1. E2 values first increased significantly at the stage of endogenous vitellogenesis and remained high during exogenous vitellogenesis. E1 levels were lower values than E2 (less than 300 pg ml^–1), but they increased at the beginning of exogenous vitellogenesis. Estrogens concentrations followed similar pattern to Vg and were significantly correlated. Mean levels of T were mostly lower than 1 ng ml^–1. They followed a pattern similar to that of E2 except for a further increase observed at the stage of final maturation. T and E2 levels were significantly correlated. The concentration of 11KT did not change significantly. The levels of 17,20-P ranged between 0.22 and 1.22 ng ml^–1 but changes were not related to gametogenesis. In males, the concentrations of T and 11KT fluctuated significantly during the sexual maturity stages, showing a similar pattern and were significantly correlated to GSI changes. T levels increased during spermiogenesis and spermiation stages to reach about 3 ng ml^–1. 11KT levels stayed about half those of T. The levels of estrogens showed no significant changes. Level of 17,20-P showed no significant variation related to male maturity. Results are discussed in relation to changes in plasma steroid levels during gametogenesis of other multiple spawner species.     

Growth performance,osmoregulatory and metabolic modifications in red porgy fry,Pagrus pagrus,under different environmental salinities and stocking densities

The effects of two different environmental salinities [brackish water (BW), 12‰; sea water (SW), 39‰] and initial stock densities [low (LD), 1.0 g L^?1; high (HD), 2.0 g L^?1] on growth, osmoregulation, stress and energy metabolism of the fry Pagrus pagrus were investigated over a period of 45 days. Pagrus pagrus (n=80, 5.51 ± 0.25 g mean initial body weight) were randomly divided in eight groups. Growth, weight gain and specific growth rate increased in BW‐acclimated fish compared with SW‐acclimated fish. No differences were observed between the two stock densities tested at either environmental salinity. Plasma osmolality was lowest in BW‐acclimated specimens, but the stock density had no effect on this parameter. Branchial Na⁺,K⁺‐ATPase activity was positively correlated with environmental salinity, but unaltered at the renal level. Plasmatic parameters were enhanced by salinity and stocking conditions. At the hepatic level, triglyceride values were enhanced in BW‐acclimated fish maintained at LD. Muscle metabolites (glycogen, glucose and lactate) increased in BW‐ compared with SW‐acclimated fish; stock density had no influence. Our data suggest that changes in metabolic parameters could be correlated with the higher growth rates observed in P. pagrus acclimated to BW, while no significant effects due to the stocking density used were observed.     

Treatment of Microcotyle sp. (Monogenea) on the gills of cage-cultured red porgy,Pagrus pagrus following baths with formalin and mebendazole

Two bath treatment trials for the control of the monogenean Microcotyle sp. in the gills of cultured red porgy, Pagrus pagrus using formalin (200 ppm for 1 h) and mebendazole (400 ppm for 1 h) were conducted. Formalin was very effective removing all the parasites from the gills of the infected fish while mebendazole produced no significant result. Based on the results of the trial, formalin was selected for the treatment of a large population of heavily infected red porgies held in sea cages. The results of the treatment are discussed.     

Effect of feeding delay after self‐feeder activation on growth performance and feeding behaviour of red porgy (Pagrus pagrus L. 1758): application to submergible cages with surface hopper

Red porgies (Pagrus pagrus) of 20.6±1.5 g mean weight were reared in tanks under four delayed self‐feeding conditions (0=control, 7, 30 and 90 s) after rod activation. The time intervals were chosen to simulate the time taken for a commercial pelleted feed to transit the tube linking a surface‐mounted feed hopper and the depth at which a cage might be submerged. Daily feeding rate, growth, food conversion index and condition factor were quite similar but fish behaviour differed among conditions. Fish remained close to the feeding point in the first two treatments (0 and 7 s delay) but foraged more widely in the remaining treatments (30 and 90 s delay), only reconsolidating around the feeding point a few seconds before feed was released. Results are discussed in relation to learning capacity and adaptation of fish to feeding behaviour flexibility, in order to develop reliable self‐feeding systems suitable for submerged cages.     

Installation,operation and evaluation of a submerged cage at 45M depth in crete for the rearing of red porgy Pagrus pagrus

Socio‐economic factors related to management of costal zones and biological parameters related to species‐specific requirements set limitations for the rearing location of net‐pen sea cages. A cage was designed and installed at 45 m depth based on the REFA Tension Length Cage technology at the cage facility of the Hellenic Centre for Marine Research in Crete. The cage‐net module consisted of a net‐pen contained within a hexagonal rigid frame to maintain the shape of the net. Vertical beams of the frame glider on six tension legs were placed in a circle. The operation of the cage was evaluated using a group of red porgy (Pagrus pagrus) for a rearing period of 4 months whereas a second group reared in a surface net‐pen cage served as control. Growth performance was similar between the experimental groups. The rearing method significantly affected fish skin pigmentation. Individuals reared in the submerged cage had a brighter skin colour and lower skin melatonin content compared with the ones reared in the surface cage. The presented submerged cage may provide a feasible alternative for rearing fish in exposed areas and particularly species requiring specific biological conditions, such as narrow temperature ranges and low light intensity.