Development of the common dentex (Dentex dentex) eggs in relation to temperature

Embryonic development of common dentex (Dentex dentex) was investigated at nine different constant water temperatures (8°C, 10°C, 12°C, 14°C, 16°C, 18°C, 20°C, 22°C and 24°C). The observed effects were compared using of regression analysis. Constant water temperatures between 12°C and 18°C were found to support successful embryonic development. A negative relationship between the rate of embryonic development and incubation temperature was observed. While embryonic development was completed within this range (12°C–18°C), there was no cell division at water temperatures of 8°C and 24°C. Total mortalities were observed at the 128 cleavage stage at a temperature of 10°C, and after the beginning of gastrulation at 20°C and 22°C.     

The effect of light intensity on early life development of common dentex Dentex dentex (L. 1758) larvae

The effects of different levels of illumination on common dentex Dentex dentex (L.) larvae have been examined. Illumination affected the relationships between total length and digestive tube length, total length and oil globule volume, and total length and yolk sac. The relationship between total length development and yolk sac utilization in dark and light conditions was found to be significant (P<0.05). However, it was determined that the relationship between digestive tube development and oil globule absorption was not significant (P>0.05). Besides, there is no effect of different light intensities on absorption of yolk sac and oil globule and digestive tube development (P>0.05). In addition, total length development was not significant (P>0.05). Survival rate was found to be not significantly different in dark conditions (P>0.05).     

Assessing the spawning season in common dentex (Dentex dentex) using microsatellites

A set of five variable microsatellite markers was used for the genetic characterization of two common dentex ( Dentex dentex ) broodstocks from an experimental hatchery and for the screening of the egg batches spawned during the complete spawn season in 2006. After parentage assignment, simulations were performed for the hatchery 1 broodstocks; the microsatellite set correctly assigned over 93% of descendants to parents when one parent was known and 80% when neither parent was known. Of the 261 eggs that the DNA was correctly amplified from, 254 were successfully assigned to a parental couple. More than one female or male took part (at the same point in time) in the spawning season. However, we observed 'dominant' individuals that produced high proportions of the descendants (unequal reproductive success) in all the sampling periods, although the contributions of these dominant individuals could change over time. This phenomenon reduces the effective breeding numbers, and could lead to inbreeding if this factor is not taken into account when the next parental generation is obtained. This factor should also be considered in aquaculture selection programmes because the dominant breeders may not be the most interesting individuals for commercial or productive schemes.     

Egg quality determination based on the shape of the lipid vesicle in common dentex, Dentex dentex

The shape of the lipid vesicle (the ratio between the maximal and the minimal diameter and the shape coefficient) of the eggs of Diplodus puntazzo and Sparus aurata is correlated with larval survival rate (%) and can be used as an egg quality marker as reported previously by Lahnsteiner and Patarnello (2005) . In the present study, this method for egg quality determination has also been validated for common dentex, Dentex dentex, after introducing several modifications in the calculation and evaluation procedure in order to obtain higher sensitivity. In the present study, it could be demonstrated that the shape of the lipid vesicle is not only correlated with the hatching rate but also with the survival rate of 1, 2 and 3 days post hatch larvae and therefore it can also be used as an indicator of larval viability at early stages of development.     

Dietary intake of probiotics and maslinic acid in juvenile dentex (Dentex dentex L.): effects on growth performance, survival and liver proteolytic activities

Two feeding trials were carried out to evaluate the efficiency of probiotics and maslinic acid, on growth and survival of juvenile dentex; liver proteolytic activities were also investigated in the second trial. For experiment 1, triplicate groups were fed six diets with two probiotics (Bacillus toyoi, T, and B. cereus, E) at increasing levels (0.5, 1 and 2 g kg^?1 diet) and a control diet. Growth and feed conversion were not significantly influenced by the probiotics. The diet T1 produced the lower mortality, whereas diet E1 rendered the higher mortality. It was concluded that no significant effects on growth and survival were found following the addition of two kinds of probiotics to dentex diets. However, the diet E0.5 showed a tendency to ameliorate the growth and feed utilization of the diet. In a second trial, triplicate groups were fed four diets with increasing levels of maslinic acid (0, 20, 40 and 80 mg kg^?1 diet). Growth of fish given diets with the highest level of maslinic acid (D80) was slightly but not significantly lower than those from the other groups. Furthemore, mortality of fish fed diet D40 was the lowest. Changes in liver proteasome and endoprotease activities measured on sodium dodecyl sulphate (SDS)/gelatin gels were also detected in a dose‐dependent manner. It was concluded that a dietary maslinic acid at a level of 80 mg kg^?1 diet seems to be too high for juvenile dentex to maintain a maximal growth and survival rate.     

Effects of dietary arachidonic and eicosapentaenoic acids on common dentex (Dentex dentex Linnaeus 1758) larval performance

Oily emulsions containing constant levels of total fatty acids (FAs) and varying eicosapentaenoic acid (EPA) and arachidonic acid (ARA) levels were used to enrich rotifers. Common dentex larval survival and growth were compared between groups fed different enriched live prey. Growth, survival rate, and lipid composition of larvae suggest that feeding common dentex in the first 15 days posthatching with 2.5–3% EPA, 6–8% docosahexaenoic acid (DHA), and DHA/EPA ratio of 2.0–2.5 is sufficient to fulfill their EPA requirements. Higher amounts of dietary EPA did not result in any significant improvement in growth or survival. EPA requirement during this period of larval development does not seem to be as critical as other FAs during the first 15 days of common dentex larval development, but it does not exclude its essentiality later in development. In the case of ARA, nutritional requirements are low compared to other marine finfish species, with the upper limit of this essential FA being around 2% of total FAs provided in the live prey composition.     

Characterization and functional properties of digestive proteases in two sparids; gilthead seabream (Sparus aurata) and common dentex (Dentex dentex)

Digestive proteases present in two sparids, seabream (Sparus aurata) and common dentex (Dentex dentex), have been characterized using both biochemical and electrophoretic techniques. Although optimum pH and temperature for maximum activity of both acid and alkaline proteases were similar in the two species, important differences in total activity, as well as in thermal and pH stability were found. Specific inhibitors and SDS-PAGE zymograms were used to clarify such differences. Evidences support the existence of a more active and complex protease set in dentex. Results are discussed from the perspective of their application to the formulation of feeds for each species.     

The rearing of common dentex Dentex dentex (L.) During the hatchery and on-growing phases

In the current paper we present the rearing methodology of common dentex (Dentex dentex), a promising candidate for Mediterranean marine aquaculture, from the larval to the on-growing phase. The results are discussed in terms of the efficiency of rearing methodology followed, for the mass production of the species.     

Effect of larval and prey density, prey dose and light conditions on first feeding common dentex (Dentex dentex L.) larvae

Initial larval stocking density, prey density, daily prey ration and light conditions (light intensity and photoperiod) were tested for common dentex larval rearing under experimental conditions. Experiments continued until the first peak of larval mortality. The best results in larval survival were obtained with an initial stocking density of between 10 and 40 larvae L^?1, fed with at least 10 rotifers mL^?1, maintaining ratios of 500–1000 rotifers larva^?1, with one or two adjustments of prey density per day. The use of more than 2000 rotifers larva^?1 or three daily adjustments of live prey density had negative effects on larval survival. The best light conditions for common dentex larval rearing were found using a photoperiod of 24 h L:0 h D and an intensity of at least 3.4 μmol m^?2 s^?1.     

Stress-related physiological responses to handling in common dentex (Dentex dentex Linnaeus, 1758)

The effects of handling on haematocrit, haemoglobin, plasma glucose, lactate and total amino acids, liver glycogen and hepatic activity of fructose 1,6‐bisphosphatase (FBPase) and alanine aminotransferase (AAT) were investigated in common dentex (Dentex dentex Linnaeus, 1758). A total of 42 fish (50‐g weight) were subjected to handling (netting and shaking in the air for 45 s). Six fish were sampled at 0, 0.5, 1, 2, 4, 8 and 12 h after handling. Six unstressed fish were also sacrificed at each time and used as controls. Handling resulted in a rise in plasma glucose and lactate levels, as well as in enhanced hepatic FBPase and AAT activities. In most cases values returned to control levels 8 h after handling. There were no significant changes in haematocrit, haemoglobin, plasma total amino acids and liver glycogen levels as a consequence of handling. The results suggest that dentex is rather tolerant to handling; apparently, sensitivity to handling is not the main factor responsible for the low survival exhibited by this species in aquaculture.     

Isolation of a highly pathogenic Vibrio pelagius strain associated with mass mortalities of turbot,Scophthalmus maximus (L.), larvae

A bacterial strain, characterized as Vibrio pelagius (Hq 222), was isolated from a turbot, Scophthalmus maximus (L.), larvae mass mortality in a commercial fish farm in Spain. Turbot larvae, post-larvae (0.2 g) and juveniles (5 and 15 g) were experimentally infected. The bacterium appeared to be very virulent for larvae and post-larvae, LD50 being < 5 bacteria mL(-1) for larvae 1 week post-infection and 3.9 x 10(5) bacteria mL(-1) in post-larvae at day 12 post-infection. The bacterial strain was recovered in pure culture from the internal organs of infected fish. Histological lesions in post-larvae exhibited swelling and necrosis of gill secondary lamellae, sloughing of intestinal mucosa and necrosis of haematopoietic tissue in the kidney. Vibrio pelagius (Hq 222) was able to grow in sterile sea water when incubated at room temperature or at 15 degrees C. Vibrio pelagius (Hq 222) was more adherent to the turbot cell lines TV-1 and TF than Escherichia coli. In both cell lines, the number of adhered bacteria increased with incubation time.     

Changes in lipid content, fatty acid composition and lipid class composition of eggs and developing larvae (0–40 days old) of cultured common dentex (Dentex dentex Linnaeus 1758)

Total lipid content, fatty acid (FA) composition and lipid class composition of common dentex eggs spawned at different times and larvae reared under different culture conditions until 40 days posthatch (dph) were analysed to get a general pattern of lipid composition during larval development. Two groups of larvae were kept under starvation to compare their FA composition with that obtained from normally fed larvae. To compare FA use or accumulation during larval development, results were grouped according to the developmental stage of the larvae instead of age in days posthatch. Saturated and monounsaturated FAs decreased along larval development, while polyunsaturated fatty acid (PUFA) content increased. The ratio of docosahexaenoic acid (DHA)/eicosapentaenoic acid shifted from 4 to 5 in early developmental stages to lower than 1 after metamorphosis. Arachidonic acid levels remained constant along larval development. Larvae kept 6 days under starvation consumed most of their n-3 PUFA while conserving the DHA to values at day 0. The results presented here are useful for the design of nutritional experiments, because there were differences detected in terms of lipid and FA composition between developmental stages with higher differences mainly found in first-feeding larvae and early developmental stages.     

An aerolysin-like enterotoxin from Vibrio splendidus may be involved in intestinal tract damage and mortalities in turbot, Scophthalmus maximus (L.), and cod, Gadus morhua L., larvae

Vibrio splendidus is a pathogen that can cause major losses during the early stages of larval turbot rearing when live feed (rotifers or Artemia) is used. As haemolytic bacteria have often been associated with larval rearing losses, we studied the role of the V. splendidus haemolysin in infection of larvae. From a bank of over 10,000 transposon mutants of V. splendidus, two different types of haemolysin-negative mutants were obtained. Both had lost virulence for larval fish, and immunohistochemistry showed that the transposon mutant studied colonized the turbot larval intestinal tract at a similar level to the wild-type organism but did not cause damage or signs of enteritis found with the wild-type organism. One transposon insertion site was located within a gene with high homology to aerolysin, the cytolytic toxin produced by several Aeromonas spp. The haemolysin, which we have termed vibrioaerolysin, had properties similar to aerolysin and osmotic protection studies showed that it formed pores in the membranes of erythrocytes of similar diameter to those of aerolysin. The Tn10 insertion site of the second transposon mutant was in an adjacent ToxR-like gene, suggesting that this might control expression of the vibrioaerolysin. The gastroenteritis caused by Aeromonas spp. in humans is considered to be due to production of aerolysin causing cyclic AMP-dependent chloride secretion in cells of the gastrointestinal tract. Damage to the intestinal tract of marine fish larvae could occur in a similar way, and it is possible that several Vibrio spp. found in the developing bacterial flora of the larval fish gut can secrete aerolysin-like toxins leading to death of larvae in the early rearing stages. Routine bacteriological screening on blood agar plates of live feed is recommended with measures to reduce the concentrations of haemolytic bacteria in rearing systems.     

Variations in Bacterial Community of Rearing Water and Gut of Common Dentex,Dentex dentex (Linnaeus 1758), Larvae Using Three Microalgae Management Approaches

Microorganisms present in the rearing water colonize the gut of first feeding larvae and represent the first barrier against opportunistic pathogens. The aim of the experiments presented herein was to standardize a protocol for the management of rearing water and microalgae suitable for the larval rearing of common dentex. In Experiment 1, bacteria–algae interactions were tested using a monospecific microalgal community, Tetraselmis chuii, suitable for nutritional experiments and with known antibacterial activity. In Experiment 2, the evolution of the bacterial community and larval performance (growth and survival) were monitored daily, in three conditions: (1) mature water: T. chuii was added 5 d before the rearing of common dentex larvae, (2) green water: T. chuii was added 1 d before, and (3) clear water: no T. chuii addition. The results show the influence of the presence of T. chuii on the evolution of the bacterial communities, both in terms of bacterial density and morphology, and indicate green water is the most suitable water treatment for management of larval rearing for common dentex.     

Protein sparing effect of dietary lipids in common dentex (Dentex dentex): A comparative study with sea bream (Sparus aurata) and sea bass (Dicentrarchus labrax)

The common dentex (Dentex dentex) is a sparid fish which is considered a suitable candidate for Mediterranean aqua-culture. A seven-weeks feeding trial was conducted over the summer period with common dentex, sea bream (Sparus aurata) and sea bass (Dicentrarchus labrax). All three species were fed to visual satiety with two practical diets with varying crude protein and crude fat levels (55 % protein, 9 % lipids; 46 % protein, 17 % lipids). The initial body weights were 8.2, 11.6 and 17.3 g for common dentex, sea bass and sea bream, respectively. In all cases, specific growth rates were not significantly affected by dietary treatment, but they varied among species (sea bass 1.7–1.8 %; sea bream 2.1–2.3 %; common dentex 3.1–3.2 %). When comparisons between fish species were made, we observed that the increase in growth rates was linked to a decrease of the whole body fat content, as a consequence of a greater utilisation of dietary lipids as energy substrates. Furthermore, in common dentex, the 17 % lipid diet was able to improve protein retention, but this diet effect was less significant in sea bream and sea bass. In contrast to sea bream, the 17 % lipid diet did not up-regulate plasma GH levels in common dentex, which provides additional evidence for a more efficient utilisation of dietary lipids.     

Viral nervous necrosis (VNN) associated with mass mortalities in cage-reared sea bass,Dicentrarchus labrax (L.)

Mass mortalities of the European sea bass, Dicentrarchus labrax (L.), occurred in different ongrowing units in Greece. A presumptive diagnosis of viral nervous necrosis (VNN) was made on the basis of the light microscopic observations of a vacuolating encephalopathy and retinopathy. Positive peroxidase reaction with VNN antiserum confirmed nodavirus as the causative agent. Usually recorded in larvae and juvenile stages of different marine fish species, VNN is described in adult sea bass and sea bass reared in floating cages in the Mediterranean for the first time. The horizontal transmission of the disease has been strongly suspected. Histological analyses clearly demonstrate that cells other than nerve cells are also infected. Some epidemiological aspects of the disease are described and their implications for the establishment of prophylactic guidelines are discussed.     

创伤弧菌、溶藻弧菌外膜蛋白特性的比较研究

对用Sarkosyl法分离的创伤孤菌、溶藻弧菌、副溶血弧菌、鳗弧菌的外膜蛋白进行了初步的比较分析.这4种弧菌外膜蛋白的SDS-PAGE和Western blotting的图谱有相似性亦有差异.SDS-PAGE显示,4种弧菌中除副溶血弧菌外均能分离到47、38 ku的外膜蛋白;创伤弧菌和溶藻弧菌存在4种共同的外膜蛋白,大...     

Reaction of common carp (Cyprinus carpio, L.) to oxygen deficiency in winter as an example for the suitability of radio telemetry for monitoring the reaction of fish to stress factors in pond aquaculture

The reaction of common carp (Cyprinus carpio L.) in an ice‐ and snow‐covered pond to depletion of dissolved oxygen (DO) in winter was recorded using radio telemetry. The carp moved from the deeper parts of the pond to the shallow areas near the tributary in search of better oxygenated water. Evidence was found that feeding occurred until December but ceased with the oxygen crisis. After the carp farmer started aeration to improve the oxygen supply, carp returned towards the deeper parts of the pond. The gross activity was significantly correlated with DO (P<0.001) and was not influenced by water temperature (P>0.05). The clear documentation of the carp's reaction to oxygen depletion suggests, that radio telemetry may be useful for investigating the effects of various stress factors on fish in pond aquaculture.     

The effect of dietary inulin on aerobic bacteria associated with hindgut of Arctic charr (Salvelinus alpinus L.)

The primary aim of the present study was to evaluate the population level of adherent (autochthonous) aerobic and facultative anaerobic bacteria in the hindgut of healthy Arctic charr (Salvelinus alpinus L.) fed dextrin or inulin. This was assessed by the dilution plate technique, and visualized using both transmission and scanning electron microscopy. A population level of 4.8 × 10⁵ adherent bacteria per gram wet mass was found in the hindgut of fish fed a casein‐based diet supplemented with 15% dextrin. However, substituting dextrin with 15% inulin reduced the bacterial population level in the hindgut (3.56 × 10⁴). A total of 217 bacterial isolates were identified by key phenotypical and biochemical characteristics. In addition, 22 strains were also identified by partial sequencing of the 16S rRNA gene. The composition of bacteria colonizing the hindgut of Arctic charr fed dextrin was dominated by the genera Staphylococcus, Pseudomonas, Micrococcus, Psychrobacter glacincola and Streptococcus. However, bacteria colonizing the hindgut of fish fed inulin were dominated by Gram‐positive bacteria of the genera Staphylococcus, Streptococcus, Carnobacterium and Bacillus. While Carnobacterium divergens‐like strains were isolated from charr fed dextrin, Carnobacterium maltaromicus‐like strains were isolated from the hindgut of fish fed inulin. Electron microscopical analysis of hindgut regions confirmed traditional culture‐based microbial analysis as fewer bacterial cells were observed between microvilli and associated with the surfaces of enterocytes of fish fed inulin rather than dextrin.