Gonadal maturation in the blackspot seabream Pagellus bogaraveo: a comparison between a farmed and a wild broodstock

The blackspot seabream Pagellus bogaraveo (Brünnich, 1768) has been regarded as a possible alternative to traditionally cultured Mediterranean species such as seabream and seabass, due to its high market value and good adaptation to captivity. Broodstock establishment and management represent the first step towards reliable production of eggs and fry, which is required to develop aquaculture of this new species. Two different broodstocks were tested for gonadal maturation and spawning, one constituting of wild fish caught as juveniles and reared in tanks until sexual maturity (4 years), and one assembled from wild adult fish caught during or just before the reproductive season. All fish were maintained under the same rearing conditions and fed the same diet. Gonadal stripping and biopsies were performed weekly to monitor maturation in both males and females. Ovarian samples were staged for maturity on the basis of follicular diameter and migration of germinal vesicle. Sperm samples were tested for density (number of spermatozoa ml⁻¹) and motility. The fish reared in captivity reached ovarian maturity during the breeding season of the wild stock. Eggs were obtained by stripping from both farmed and wild specimens, but appeared degenerated as a result of being retained too long in the ovarian cavity due to the absence of spontaneous spawning. Spermiation was prolonged in the farmed fish, but appeared to be blocked in the wild breeders after first sampling. However, the sperm was very viscous and the motile spermatozoa did not exceed 10%.     

Hyperplastic and hypertrophic growth of lateral muscle in blackspot seabream Pagellus bogaraveo from hatching to juvenile

To understand better the growth mechanisms in the economically important fish Pagellus bogaraveo, in terms of muscle fibre hyperplasia v. hypertrophy, the lateral muscle of this fish was studied morphometrically from hatching to juvenile comparing rostral and caudal locations. Fish were sampled at 0, 5, 23, 40, 70, 100, 140 and 180 days. Fibre types were first identified by succinate dehydrogenase (SDH) and immunostaining with a polyclonal antibody against fish slow myosin (4–96). Morphometric variables were then measured in transverse body sections, at both post‐opercular and post‐anal locations, to estimate the following variables: total muscle area [A (muscle)], total fibre number [N (fibres)], fibre number per unit area of muscle [N_A(fibres, muscle)] and cross‐sectional fibre area [ (fibres)] of the two main muscle fibre types (white and red). Overall, growth throughout the various stages resulted from increases both in the number and in the size of muscle fibres, paralleled by an expansion of the [A (muscle)]. Nonetheless, that increase was not significant between 0–5 days on one hand and 100–140 days, on the other hand. On the contrary, the [N_A(fibres, muscle)] declined as the body length increased. Analysis of the muscle growth kinetics suggested that, within the important time frame studied, hyperplasia gave the main relative contribution to the increase of white muscle [A (white muscle)], whereas red muscle [A (red muscle)] mainly grew by hypertrophy, with both phenomena occurring at a faster pace posteriorly in the body. Finally, when comparing rostral and caudal locations, a greater [N (fibres)] and [A (muscle)] of the posterior white and red fibres were the consistent features. It was also observed that the proportion of the cross‐sectional area of the myotomal muscle comprised of white muscle was greater in the anterior part of the fish.     

Muscle development in cultured blackspot seabream Pagellus bogaraveo: preliminary histochemical and immunohistochemical data on the fibre types

We studied muscle ontogeny and fibre type characteristics in the blackspot seabream, a new species for commercial aquaculture. Myosin ATPase and SDH histochemistry and immunohistochemistry were tested at different ontogenetic stages, using a panel of antibodies to myosin isoforms and parvalbumin. In general, deep white muscle was parvalbumin‐positive, and superficial 'red' muscle was parvalbumin‐negative at all ages examined. At 6 days of age (transition from endogenous to exogenous feeding) three layers of muscle fibres were observed with different antimyosin reactivities: superficial monolayer, presumptive slow red (present only as a small group of fibres adjacent to the lateral line nerve), and presumptive fast‐white (forming the bulk of the muscle). The superficial monolayer and presumptive slow fibres were positive for SDH. At 60 days of age (transition from live to artificial feeding) an additional fibre type was identified: a typical 'pink' or intermediate layer. In juveniles, the axial muscle consisted mainly of fast white fibres covered by a slow‐red layer and between them a pink layer. Surprisingly, the red layer could be resolved into two distinct types by myosin immunostaining. Red fibres were also present along the horizontal septum, near the notochord. Both red and white muscle layers showed a mosaic appearance, which was confirmed by ATPase reaction. The work was financed by British Council, CRUP, and FCT (PhD Grant SFRH‐BD‐14068–2003).     

Muscle differentiation in blackspot seabream (Pagellus bogaraveo, Brunnich): histochemical and immunohistochemical study of the fibre types

The aim of this work was to gain insights into the mechanism of muscle differentiation and growth in Pagellus bogaraveo, by studying muscle fibre phenotypes identified by immunohistochemistry. At hatching, several layers of deep fast-white fibres were covered by a superficial fibre monolayer. At 5 days, slow-red fibres appeared near the lateral line nerve. At 40 days, the intermediate-pink muscle became visible, and in the slow-red and fast-white muscle layers transitions from larval myosin isoforms to the isoforms typical of adult muscle occurred. Between 70 and 100 days, small fibres with a distinct ATPase profile appeared throughout the fast-white muscle, marking the onset of “mosaic” hyperplasia. The myosin of the original superficial monolayer fibres underwent two myosin transformations, before being slowly replaced by an adult slow-red isoform. In juveniles and adults, the slow-red muscle layer could be resolved into two distinct types. The analysis of fibre phenotypes indicated that post-larval muscle growth occurred by two distinct stages of hyperplasia. This study offers a basis for further comparative and experimental studies with this economically relevant species, namely for identifying factors influencing its muscle growth dynamics and disclosing underlying mechanisms.     

Influence of dietary protein level on growth performance and body composition of juvenile blackspot seabream, Pagellus bogaraveo(Brunnich, 1768)

Blackspot seabream Pagellus bogaraveo is a potential candidate species for southern European aquaculture. A 12‐week feeding trial was then conducted to evaluate the effect of dietary protein level on growth performance, body composition and feed use efficiency of blackspot seabream juveniles (22·7 g each). Five isolipidic diets (12·5%) with graded levels of protein (20, 30, 40, 50 and 60%) were hand‐distributed, twice a day, to duplicate groups of fish (120 fish per tank), until satiation. Growth (DGC), survival and feed utilization (FCR) were recorded through monthly sampling. Growth performance was significantly affected by dietary protein level ( P  < 0·05), and fish fed at the 50% protein level exhibited the highest final body mass. Feed conversion ratio decreased with increasing levels of dietary protein, fish fed 60% protein being the most efficient. No significant differences were found in whole body composition among treatments ( P > 0·05). Financed by FCT (Project POCTI/CVT/39239/2001, PhD Grant SFRH‐BD‐14068–2003)     

Spatial distribution and life‐history aspects of blackspot seabream Pagellus bogaraveo (Osteichthyes: Sparidae)

Spatial distribution and life history aspects of Pagellus bogaraveo in the eastern Ionian Sea were investigated using the data from 13 different studies carried out in the area from 1983 to 2010. The spatial patterns of the abundance, biomass and mean size showed that the species inhabits the shallow waters of the shelf (<170 m depth) as juveniles up to a certain size (<180 mm total length, L_T), moving to deeper waters of the slope (mainly 400–500 m depth) as adults. The spatial pattern of abundance indicated a continuous distribution of the species in deep waters, with hot‐spot areas of high values, whereas in shallow waters distribution was more discontinuous, with higher concentrations of juveniles in estuaries and brackish waters. The study of biological aspects of the species revealed (1) a difference in the increase in mass between males and females, (2) protandrous hermaphroditism, accompanied by the presence of primary females and males that do not change sex, (3) a sex ratio in favour of females >250 mm L_T, (4) the presence of hermaphrodites between 200 and 370 mm, (5) a long reproduction period from June to March, (6) a size at first maturity around 300 mm and (7) a diet composition of adults based mainly on fishes, and also on opportunistic behaviour in the food scarce environment of deep waters. The results suggest that the species' distribution and feeding strategies are the most appropriate for the oligotrophic eastern Ionian waters and that these conditions result in smaller sizes of the species in the east Mediterranean Sea compared to the west basin and the east Atlantic Ocean, with implications for the growth and reproductive biology of the species.     

The effect of elevated hydrostatic pressure upon selected biomarkers in juvenile blackspot seabream Pagellus bogaraveo in a 14 day‐long experiment

Hydrostatic pressure elevated to 500 kPa for 14 days was found to affect hepatic 7‐ethoxyresorufin‐O‐deethylase (EROD), oxidized protein (POx), protein yield and branchial Na⁺–K⁺‐ATPase. No effect on glutathione‐S‐transferase (GST), superoxidase dismutase (SOD), catalase (CAT), lipid peroxidation (LP), acetylcholinesterase (AChE), butyrylcholinesterase (BChE), condition factor (K) and hepato‐somatic index (I_H) was encountered.     

Dynamics of black spot sea bream (Pagellus bogaraveo) mean length: evaluating the influence of life history parameters, recruitment, size selectivity and exploitation rates

Stochastic simulations were used to evaluate the influence of recruitment pattern (log‐normal, decreasing), size selectivity (normal, logistic model) and fishing mortality pattern (abrupt, continuous increase in fishing mortality) on the evolution of mean length and the dispersion of mean length for a relatively long‐lived deep‐water species, the black spot sea bream (Pagellus bogaraveo). An abrupt increase in fishing mortality resulted in mean size decreasing and stabilizing at a lower level while a steady increase in fishing mortality caused the continuous decrease in mean size that has been reported for many long‐lived species. Decrease in mean size was greatest for logistic model simulations and for cases where fish were susceptible to capture at a small size. Logistic selectivity, with decreasing recruitment and increasing fishing mortality over time, resulted in mean length and variability in mean length trends similar to that observed for the Strait of Gibraltar fishery. Furthermore, it was found with the declining recruitment that moderate increases in fishing mortality can result in significant decreases in mean length. Given the importance of mean size as an indicator of the state of a resource, these simulations are a useful alternative or complement to standard fisheries assessment methods, helping to provide information on exploitation patterns and rates that can be used for conservation and management.     

Rice protein-concentrate meal as a potential dietary ingredient in practical diets for blackspot seabream Pagellus bogaraveo: a histological and enzymatic investigation

Field and laboratory studies were conducted to evaluate the intestinal responses to partial replacement of fish meal with rice protein concentrate (RPC) in practical diets for blackspot seabream Pagellus bogaraveo . Two experimental diets were formulated to be isoproteic and isoenergetic with an increasing level of RPC (20 and 35%, respectively) and were tested against a fish meal-based control diet (RPC0). The diets showed similar features for growth performances and both intestinal histology and digestive enzymes. This study confirmed that RPC does not induce intestinal mucosa alterations in this fish. The dietary RPC supplement caused a significant increase in trypsin activity, whereas lipase activity was reduced.     

Complete mitochondrial genome of the blackspot seabream, Pagellus bogaraveo (Perciformes: Sparidae), with high levels of length heteroplasmy in the WANCY region

The complete mitochondrial genome sequence of the blackspot seabream, Pagellus bogaraveo, was obtained using the long PCR/cloning method. The total length of the mitogenome was 16,941 bp, and had a gene content (13 protein-coding, two ribosomal RNAs, and 22 transfer RNAs) and organization similar to those observed in most other vertebrates. Nevertheless, two main features in the WANCY region revealed as unique in P. bogaraveo mitogenome. First, the O(L) was disrupted by the insertion of a 66 bp long element that was determined to be a tRNA(Cys) pseudogene; second, the existence of high levels of length heteroplasmy, both intra-and inter-individuals, as a result of sequence duplications and deletions. The tandem duplication and random loss (TDRL) model as well as recombination are proposed to account for the length heteroplasmy and gene rearrangements in the P. bogaraveo WANCY region.     

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 south‐west 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 (L₅₀) for all sexes and undetermined individuals combined was 18.33 cm total length (TL), with no significant differences between males and females. The estimated L₅₀ 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.     

The effect of exercise and captivity on energy partitioning, reproductive maturation and fertilization success in adult sockeye salmon

Releasing a population of adult Fraser River sockeye salmon Oncorhynchus nerka from an energetically more expensive non‐feeding natural migration (1089 km, natal groups) and allowing them to sexually mature in either a moderate flow (captive exercise group) or a no flow (captive non‐exercise group) environment resulted in significant differences in body energy densities (MJ kg⁻¹) among groups (ranked: initial natural migrator > non‐exercise > exercise > natal arrivals). Similarly, per cent body lipids were significantly lower in exercise females compared with non‐exercise females, experimentally confirming the central role lipid catabolism has in powering swimming. Although restricting exercise did result in greater body energy reserves at spawning, this did not result in a reallocation of energy to reproduction, as fecundity, egg size and gonad composition were unchanged among the groups. Furthermore, non‐exercise females had delayed maturity, lower egg deposition rates, and were more likely to die prior to egg ovulation than exercise females and natal spawners. Eggs from captive exercise adult females were more likely to survive to the eyed stage than eggs from captive non‐exercise females, but both captive groups had significantly lower egg fertilization success than natal spawners suggesting that confinement stress played a role in some of results presented. There were no significant differences in plasma levels of glucose, lactate, cortisol and reproductive hormones among the groups, but correlations among acute and chronic indicators of stress were significant among individuals. These results indicate that exercise during the late stages of maturation, as might occur normally, may be required for optimizing reproductive maturation and maximizing reproductive success.     

Reproductive biology and recruitment of the white sea bream in the Azores

The life history of the white sea bream Diplodus sargus in the Azores showed a pattern consistent with digynic hermaphroditism achieving sexual maturity during the second year of life, at 16·7 cm L _T. Spawning occurred from March to June at temperatures between 15 and 17° C and the onset and duration of spawning season in the sea bream appeared to be influenced by sea water temperatures. As latitude decreased, both in the northern and southern hemispheres, the spawning season of D. sargus populations started earlier and extended longer, highlighting the potential importance of temperature to the onset and duration of reproduction in this species. Settlement took place from late May to July, and settlers remained in the nursery area for c . 2·5 months. Emigration from the nursery area to join shoals of juveniles occurred from late July to September.     

Optimization of single-strand conformation polymorphism and sequence analysis of the mitochondrial control region in Pagellus bogaraveo (Sparidae, Teleostei): rationalized tools in fish population biology

We report the isolation and sequencing of 400–550 base pairs (bp) of the mitochondrial DNA (mtDNA) control region of eight species of Sparidae (Perciformes, Teleostei). This sequence information allowed us to design specific primers to one of these species (Pagellus bogaraveo). The new set of primers was used to test a rationalized approach to study the mtDNA nucleotide variability at the intraspecific level. The single-strand conformation polymorphism (SSCP) technique was applied to detect sequence variation in two non-overlapping fragments of the control region of 32 individuals of P. bogaraveo. To assess the sensitivity of the method, the nucleotide sequence of the analysed region was determined for all the specimens. The results showed that, for one of the two fragments, SSCP analysis was able to detect 100% of the underlying genetic variability. In sharp contrast, nucleotide variation of the second DNA fragment was completely unresolved by SSCP under different experimental conditions. This suggests that the resolution power of SSCP is crucially dependent on the nature of the fragment subjected to the analysis; therefore, a preliminary test of the sensitivity of the method should be performed on each specific DNA fragment before starting a large-scale survey. A rationalized approach, combining the SSCP technique and a simplified sequencing procedure, is proposed for studying intraspecific polymorphism at the mtDNA control region in fish.     

The larval development of lateral musculature in gilthead sea bream Sparus aurata and sea bass Dicentrarchus labrax

Fibre-type differentiation of the lateral musculature has been studied in Sparus aurata (L.) and Dicentrarchus labrax (L.) during larval development. Histochemical and ultrastructural techniques show two presumptive muscle layers and two germinative zones of presumptive myoblasts. At hatching, myotomal muscle consists of a monolayer of thin undifferentiated cells near the skin (first germinative zone) overlying another mono-layer of small diameter fibres extending hypaxially and epaxially away from the transverse septum. Below this, there is a much thicker, deep layer of fibres, generally large in diameter and polygonal in shape. The presumptive myoblasts are located between these two layers of fibres in the second germinative zone. Initially, the superficial and deep muscle fibres show high and low myosin ATPase activity, respectively. Both layers grow by generating new fibres from the two mentioned germinative zones. At the end of larval life, the superficial layer changes its histochemical profile from high to low myosin ATPase activity and, at the same time, intermediate or pink muscle fibres can be observed by oxidative activity (the NADH-TR reaction). Morphometric analysis shows a significant increase in mean fibre diameter during successive ages, as shown by the Student's t-test (hypertrophic growth). Skewness and kurtosis values of fibre diameters point to the generation of a new fibre population from the germinative zones (hyperplastic growth).     

Growth, gonadal development and sex ratios of meiogynogenetic diploid sea bass

Gynogenesis showed little effect on general physiology and gonadal development in sea bass Dicentrarchus labrax . Meiogynogenetic fish showed well-developed gonads indicating low occurrence of developmental imbalances even after gynogenesis induction in this species. In addition, the proportion of sexes of meiogynogenetic sea bass was similar to the diploid controls in two independent trials, which did not deviate significantly from a 1:1 male: female sex ratio. Even considering some environmental influence on sex differentiation, as has been previously demonstrated, the fact that the proportion of sexes was similar between gynogenetic and control diploids essentially eliminates the possibility that in the sea bass the females are the homogametic sex. Although the mechanism of sex determination of this species still remains unknown, even after gynogenesis induction, the genetic mechanism of the ZW/ZZ type could probably operate in the sea bass.