Inter- and intraspecific crossbreedingexperiments were conducted to evaluate the aquaculture potentialof hybrids in the genus Dicentrarchus, focusingon fertilisation and hatching success.The experimental designconsisted of 24 controlled crosses in which individual and pooledfertilisations were made between wild D. labrax individuals(8 dams and 5 sires originating from West Mediterranean and Atlanticpopulations) and wild D. punctatus (6 sires). Threeexperiments were successively performed: (1) dams from the Mediterraneanpopulation, individually considered to have good egg quality, (2)dams from Mediterranean population, individually considered to havepoor egg quality and (3) crossbreeding using pooled eggs from theAtlantic population.In each case, batches of these eggs were fertilizedby sperm from males of the two species. Sperm cell quality (concentrationand motility) was verified before experimentation in all cases andequal numbers of sperm cells from each male were used to individually(without inter-sire competition) fertilise egg batches. Throughthe repeated artificial crosses between female common sea bass Dicentrarchuslabrax (Linnaeus 1758) and male spotted sea bass D.punctatus (Block 1792), these experiments showed that nopost-zygotic reproductive barriers exist to interspecific hybridisationbetween these two species when using D. labrax asdams and D. punctatus as sires. Phenotypically,the F1 hybrids were easily recognisable: they inherited the characteristicblack spots of D. punctatus. Furthermore, embryosurvival was significantly higher in interspecific crosses comparedwith intraspecific controls, showing increased fitness for this trait(increased performance at early life stage). Then, the experimentalbreeding design validates the observation that West Mediterraneanand Atlantic common sea bass populations are different. Indeed,the inter-population crosses (between West Mediterranean dams andAtlantic sires) also revealed increased fitness at early life stagesin comparison with the progeny of intra-Atlantic population crosses.However, these individuals were still smaller than their interspecificcounterparts. The ability to produce viable F1 hybrids will havesignificant implications for the practical improvement of sea bassaquaculture.

Batch fecundity and relative fecundity of swordfish (Xiphias gladius) in the southwestern Indian Ocean were estimated from seven gravid swordfish females (size range 127–225 cm lower jaw-to-fork length, LJFL) with unovulated, hydrated oocytes collected onboard Reunion-based (France) longline swordfish fishing vessels between December 1999 to January 2001. To investigate the spawning pattern of swordfish, we used data collected through a combination of two at sea sampling regimes. A total of 17 007 geo-located size data of swordfish were recorded during 8 years (1993-2001) and a total of 1727 (size range 75–289 cm, LJFL) swordfish gonads (1107 females and 620 males) were sampled from May 1998 to January 2001. The estimated batch fecundity ranged from 995 000 hydrated oocytes for the smallest ripe female to 4.3 millions for the largest female sampled measuring respectively 127 to 225 cm in curved length (LJFL). The relative fecundity ranged from 25 to 72 hydrated oocytes per gram of body weight. We found that batch fecundity was positively correlated with fish length and that the older/larger females have earlier and longer spawning seasons than younger/ smaller females. These findings suggested that older/larger females which are seasonally migrating in this spawning ground seem to play a major role in reproductive success of the species in producing significantly more offspring than younger females during an extended spawning season. Examination of the length-frequency date from the fishery indicatedthat the young fish are resident around Reunion and around the seamounts off Reunion Island. Our results highlight the important role of the older/larger females in the reproductive capacity of southwestern Indian Ocean stock. We discuss the potential implications of fishing the older/larger females for this stock in terms of reproduction and recruitment.

The reproductive dynamics of swordfish (Xiphias gladius) was investigated from catches of the Reunion Island-based longline swordfish fishery between 19–25 °S and 48–54 °E. Thus, 1727 swordfish (size range 75–289 cm, lower jaw-to-fork length, LJFL) were sampled on board commercial fishing vessels during the period of May 1998 to January 2001. Reproductive activity was assessed using macroscopic gonad characteristics, trends of gonadal indexes for both sexes, oocyte size-frequency distributions and microscopic investigation of oocyte development stages. Size at first maturity (L 50) for female and male swordfish was estimated in Indian Ocean for the first time. L 50 was estimated at 170 cm and 120 cm (LJFL) for females and males, respectively. Spawning occurred from October to April in the vicinity of Reunion Island where sex ratio of caught populations fluctuated seasonally. The seasonal changes in sex ratio and the incidence of larger individuals at the beginning of the spawning season provide some preliminary indications of synchronized movements of swordfish between spawning grounds and neighboring regions. Our results add significantly to knowledge about the reproductive biology of swordfish in the southwestern Indian Ocean. An overview of the available information on reproductive biology and dynamics of swordfish in Indian Ocean and eastern Pacific indicated that spawning activity is localized in discrete areas, and showed that there is incomplete information on spawning grounds in this extended area.

Sexual differentiation was studied at the histological level using a mixture of 30 families of sea bass Dicentrarchus labrax. Most of the fish (93%) differentiated into males as usually observed in farmed populations. All testes were differentiated when the males reached 12 cm and no more undifferentiated fish were found from 419 days post-fertilization (p.f.). In 28% of the males, among the biggest, sexual differentiation had already begun at 168 days p.f. (8.3–9.5 cm) and these fish started spermatogenesis in their first year of life. The other males differentiated later and remained immature at the end of their first year of life. Ovaries could be identified at the histological level from the age of 168 days p.f. (7.9–9.0 cm) and the females became significantly longer than the males from the age of 191 days p.f., i.e. during the process of ovarian differentiation. In the studied group, 62% of the males developed intratesticular oocytes. Such intersexuality had no consequence on growth rate. Intratesticular oocytes were also recorded in testes of wild males originating from Atlantic (Britain and Gulf of Gascogne) and West Mediterranean showing that juvenile intersexuality is not restricted to farmed populations but is a widespread phenomenon in sea bass.

The present work examines the usefulness of the combination of multivariate analyses and further one-way analysis of variance in the search for links between causes and effects in heterologous stimulation of spawning in fish. Stimulations with both 20 and 50 μg.kg–1 doses of [D Trp6] luteinizing hormone-releasing hormone analogue (LH-RHa) were effective in inducing and synchronizing spawning of red drum, Sciaenops ocellatus. Oocyte development monitoring using ovarian biopsy allowed treatment injections at different stages of maturation. The influence at stimulation time of female oocyte development state upon further development was shown. An interesting potential of second stimulation, leading to high quality spawns, was established and factors of success were identified. The best conditions for stimulation success were simultaneously late stage of oocyte development characterized by the beginning of oil droplet coalescence, high ovarian homogeneity quoted by the proportion of most developed oocytes in the biopsy sample, and low dose of hormone (20 μg.kg–1) for both first and second stimulations.

A sperm cryopreservation protocol adapted from turbot, was tested on sea bass using either 250-μL straws or 1.5-mL cryovials. A dilution to 1/3 in Mounib's extender and a cooling rate of −65 °C·min−1 allowed frozen sperm to recover an initial motility similar to that of fresh sperm at thawing; however, significant differences in motility (P < 0.001, n = 10 fish semen) were observed at further post-activation times, the motility decrease being faster in thawed sperm. At the experimental scale, triplicate inseminations of 2-mL aliquots (approximately 2 000 eggs) showed a significant fertility decay of thawed sperm compared to that of fresh sperm (P < 0.01, n = 12 fish semen) when a discriminating 35·103 spermatozoa to egg ratio was applied. When 70·103 and 200·103 spermatozoa per egg were provided in the same experimental conditions, no significant difference appeared between the fertilisation rates of fresh and thawed sperm. In order to validate the procedure for production or cryobank purpose, a scaled-up protocol was established. Two and 50 mL batches of eggs (approximately 2·103 and 50·103 eggs, respectively) were inseminated in triplicate using either fresh or thawed individual sperms of 5 males with 200·103 spermatozoa per egg. The mean fertility decreased by 23.5 % due to cryopreservation. This decline was explained by the loss of fertility of only one sperm, and only in large-volume conditions, probably due to the delay of use after thawing.

This paper presents some features of turbot sperm and compares them with data reported in the literature on other freshwater and marine fish. In captivity, turbot spermiation period lasts 6 months. When compared to other fish species, turbot males are characterized by low values of maximum gonadosomatic index (0.6-0.8%) and are poor sperm producers in terms of volume (0.2-2.2 ml), concentration (0.7-11.0 × 109 spermatozoa/ml) and total sperm number released per stripping (0.2-12.0 × 109 spematozoa). Compared to most freshwater species, the duration of cell movement is long (1-17 min). The spermatozoon of turbot can be considered to be of a primitive type. A very high protein content (8.8 mg/ml) is found in the seminal fluid of turbot. Increasing collection frequency of turbot males, from monthly to fortnightly and weekly stripping, results in the release of successive samples presenting decreasing semen volume and spermatozoa concentration. Similar spermatozoa concentration and total volume of semen produced per turbot, during a 2 month experimental period, are released from males submitted to an annual light and temperature cycle, compared to those receiving a 6 month contracted light and temperature schedule.

Vitellogenin (VTG) was isolated from the haemolymph of Penaeus vannamei by a three stepprocedure including ultracentrifugation, gel filtration and ion exchange chromatography. VTG wasused to raise polyclonal antibodies that were purified by ion exchange chromatography. A two stepcompetitive assay was developed in which VTG could be quantitated by its capacity to inhibit thebinding of antibody to the VTG previously adsorbed onto a solid phase. Sensitivity from equilibriumand from non-equilibrium assays was 41 and 2.3 ng/ml respectively. Estimates of within-assay andbetween-assay variabilities of standard curves were 4.2 and 9.6% respectively. The inhibition curves for dilutions of haemolymph from vitellogenic females, egg yolk extracts and purified vitellin (VTL) were parallel to the standard VTG curve, haemolymph from immature females and males showed no cross-reactivity. The antibodies directed against VTG recognize but partially VTL as shown by displacement curves. Recovery tests were near 100%. Thus the procedure was considered to be suitable for the measurement of haemolymphatic VTG. The VTG enzyme-linked immunosorbent assay (ELISA) developed in this study was validated by detecting physiological VTG changes in female shrimps after being fed squid extracts.