Off-Season Spawning of Eurasian Perch Perca Fluviatilis

This study was designed to determine the effects of the selection for body weight and of two final warming durations during an out-of-season reproductive cycle on gonad recrudescence, plasma steroid levels and spawning in Eurasian perch Perca fluviatilis males and females. Two different bodyweight classes (200–250 and 350–400 g) and two different warming periods (1 or 2 months) were studied.At the end of the chilling period, a significant effect of the selection for body weight was observed on gonad development whereas fish were the same age. According to higher levels in testosterone (12 ng mL^–1), a better development was recorded for smaller breeders (P < 0.05). A short water warming period up to 14°C (1 month v.s. 2 months) allowed higher rates of females with completed gonadogenesis (33–38% v.s. 7–8%) and spawning (31–33% v.s. 0%) (P < 0.05). Spontaneous out-of-season spawning (17 ribbons) and larvae were obtained. Relative fecundity was about 100 eggs g^–1 of female body weight. Five ribbons were partly fertilized with a rate ranging from 5 to 90%. These studies have shown that delayed spawning in Eurasian perch can be obtained by a temperature manipulation, but results are not sufficient because of low stimulation rate of females, low spawning rates and high heteroge-neity of fertilization rates. The effects of other environmental factors like photoperiod should be tested in further experiment to improve the protocol inducing out-of-season spawning production.     

Nitrogen and phosphorous budgets during a farming cycle of the Manila clam Ruditapes philippinarum: An in situ experiment

In the Sacca di Goro lagoon a farming cycle of the Manila clam (Ruditapes philippinarum) was simulated seeding young molluscs in an unexploited sandy spot. The experimental area (2100 m²) consisted of three sectors: a control (C), almost devoid of clams (∼ 1600 m^− 2, ∼ 30 ind m^− 2), a low (L) density area (400 m², ∼ 300 ind m^− 2) and a high (H) density zone (∼ 110 m^− 2, ∼ 800 ind m^− 2). Water chemistry, external freshwater nutrient loads, molluscs filtration rates, biomass, elemental composition and nutrient recycling were analysed.Clam filtration rates and light and dark fluxes of nutrients were measured with intact core incubations. Three replicate cores (i.d. 20 cm) were collected from C, L and H in April, one month after the seeding, June, August and October 2003. External loads were calculated multiplying dissolved and particulate nutrients concentration by freshwater flow from the main lagoon tributaries. Direct excretion, filtration activity of clams and particulate matter deposition resulted in significantly higher ammonium (NH₄⁺) and soluble reactive phosphorus (SRP) effluxes to the water column at L and H. For the entire farming cycle, particulate nitrogen (PN) uptake by clams from the water column was 1.7 (C), 9.1 (L) and 16.3 (H) mol m^− 2, whilst total dissolved nitrogen (TDN) fluxes were − 0.3 (C), 1.6 (L) and 6.9 (H) mol m^− 2. Particulate phosphorus (PP) uptake from the water column was 0.1 (C), 0.6 (L) and 1.0 (H) mol m^− 2, whilst total dissolved phosphorus (TDP) efflux was 0.2 (C), 0.5 (L) and 0.8 (H) mol m^− 2. At the end of the farming cycle, harvested N as mollusc flesh was negligible for C, 0.4 mol m^− 2 for L and 1.8 mol m^− 2 for H. Harvested P as mollusc flesh was negligible for C, 0.02 mol m^− 2 for L and 0.04 mol m^− 2 for H. Farmed areas seem to have a great potential for fast coupling between sedimentation (filter feeder mediated biodeposition) and benthic recycling. At the lagoon level, mollusc farming probably attenuates the export of particulate matter to the open sea. Our results show that a minor fraction of biodeposited N (∼ 6%) and P (∼ 3%) was exported as a commercial product at the end of the farming cycle, whilst a larger fraction was incorporated in the sediments or recycled as dissolved inorganic or organic forms.     

Disruption of gonadal maturation in cultured Senegalese sole Solea senegalensis Kaup by continuous light and/or constant temperature regimes

A major problem in the development of Senegalese sole Solea senegalensis intensive culture is the poor control on reproduction, in part due to the lack of knowledge on the precise role of photoperiod and temperature. Thus, gonadal maturation was evaluated by assessing the sequential changes in plasma levels of 17β-estradiol (E₂), 11-ketotestosterone (11-KT), and testosterone (T) in both female and male cultured Senegalese sole (F1 generation) exposed to various combinations of constant or naturally-fluctuating daylength and water temperature. Under natural photoperiod (NP; 36° N), exposure to constant temperature (t0; 18-20 °C) disrupted gonadal development, as indicated by a lower incidence (in comparison with naturally-fluctuating water temperature; 14-24 °C) of females at advanced maturation (from February to April: 12 vs. 33%) and running males (from February to May: 46% vs. 57%), and the reduced mean (± S.E.M.) sex steroid plasma levels (female peak E₂ levels: 2.9 ± 0.28 vs. 1.8 ± 0.3 ng ml^− 1; male peak T levels: 1.5 ± 0.14 vs. 0.9 ± 0.06 ng ml^− 1). Therefore, the onset and progression of gonadal development in this species seem to be strongly (“proximally”) influenced by fluctuating water temperature. When compared to NP and t0, exposure to continuous light (LL) under t0 significantly reduced steroid production (female peak E₂ levels: 1.8 ± 0.28 vs. 0.5 ± 0.05 ng ml^− 1; male peak 11-KT levels: 9.4 ± 1.06 vs. 5.4 ± 1.33 ng ml^− 1) and subsequently gonadal development (lower proportions of females at intermediate [46 vs. 6%] and advanced maturation [12 vs. 0%] from February to April and of RM [46 vs. 33%] from February to May). Thus, the seasonal changes of daylength would be crucial for normal gonadal development, being its cueing effects of higher magnitude than those of water temperature. The present report constitutes the first systematic study focused on the environmental control of reproductive events in Senegalese sole.     

Ammonia toxicity and its effect on the growth of the South African abalone Haliotis midae Linnaeus

The current study investigated acute toxicity to ammonia of the South African abalone, Haliotis midae, from three size classes relevant to mariculture operations, and the chronic impact of sub-lethal ammonia levels on growth of juvenile abalone.Results showed that tolerance to ammonia (at pH 7.8 and T_a = 15 °C) increases with body size (i.e. age) as indicated by 36 h LC₅₀ values: juvenile abalone (1-2.5 cm shell length) had the lowest LC₅₀ of 9.8 μg l^− 1 FAN, whereas LC₅₀ was 12.9 μg l^− 1 FAN in “cocktail”-size abalone (5-8 cm shell length). The highest LC₅₀ of 16.4 μg l^− 1 FAN was observed in “brood stock”-size animals (10-15 cm). When “cocktail”-size abalone were allowed to acclimatize to sub-lethal ammonia levels for 48 h, their ammonia tolerance increased compared with non-acclimatized abalone of the same size: LC₅₀ was 2.0 μg l^− 1 FAN higher at 14.8 μg l^− 1 FAN.Growth of juvenile abalone (1-2.5 cm shell length) during chronic exposure to sub-lethal FAN levels is inhibited: specific growth rate (SGR) is significantly reduced by 58.7% to 0.10 ± 0.03% d^− 1 (weight) compared with 0.24 ± 0.06% d^− 1 of abalone of a control group (no ammonia).The results demonstrate the negative effects of ammonia not only on survival but also on growth of farmed abalone, both impair profitability of the farming operation. The information from the present study will be helpful in determining water quality requirements in South African abalone farms.     

Effect of saponin on hematological and immunological parameters of the giant freshwater prawn, Macrobrachium rosenbergii

Giant freshwater prawns, Macrobrachium rosenbergii (17.9 ± 2.7 g), exposed to different concentrations of saponin at 0, 0.3, 0.6, 0.9 and 1.2 mg l^− 1 for 168 h were examined for osmolality, electrolyte levels, oxyhemocyanin, protein levels, acid-base balance status, total hemocyte count (THC), phenoloxidase activity, and respiratory bursts. Hemolymph oxyhemocyanin, protein, and pO₂ were inversely related to the saponin concentration. Hemolymph oxyhemocyanin, protein, pO₂, pCO₂, and pH of prawns exposed to 1.2 mg l^− 1 saponin were significantly lower than those of prawns exposed to 0.3 mg l^− 1 and control solutions. However, no significant difference was observed in osmolality or electrolyte levels of prawns exposed to different concentrations of saponin for 168 h. The THC of prawns following 168 h of exposure to 0.9 and 1.2 mg l^− 1 saponin increased, but the phenoloxidase activity decreased suggesting that the decrease in phenoloxidase activity under saponin stress was not a consequence of the increase in THC. We concluded that saponin at as low as 0.9 mg l^− 1 decreases the respiratory protein level and acid-base balance, and modulates the immune system of M. rosenbergii.     

Reproductive cycle and plasma levels of steroids in male Eurasian perch Perca fluviatilis

From April 1995 to April 1996, the annual reproductive cycle of male Eurasian perch Perca fluviatilis was studied at the Lindre Center (Moselle, France). At monthly intervals, five males (mean body weight of 133 ± 43 g and total length of 175 ± 9 mm) were caught. From sampled organs, the gonadosomatic (GSI), hepatosomatic (HSI) and viscerosomatic indexes (VSI) were calculated and plasma testosterone (T) and 11-ketotestosterone (11KT) levels were measured. After the spawning period in mid-April, GSI and HSI dropped and VSI increased to 3.8 ± 0.1 %. In September, GSI reached its maximum (8.5 ± 1.8 %). At this time, spermatocytes, spermatids and spermatozoa were abundant, whereas in June only spermatogonia were observed. During winter, GSI was stable at approximately 5 %, HSI reached its maximum (1.9 ± 0.3 %) and VSI was low (2.6 ± 0.2 %). From April to November 1995, plasma T and 11KT concentrations were low (< 0.5 ng·mL^–1). Plasma T levels increased significantly in December and reached peak levels (12.3 ± 2.1 ng·mL^–1) in January, then decreased in February and increased again until spawning in April (6.8 ± 2.1 ng·mL^–1). This second elevation could be related to the beginning of a new spermatogenic cycle. Plasma levels of 11KT increased significantly from October to February 1996 (4.9 ± 1.1 ng·mL^–1). From February to the spawning period (April 1996), plasma 11KT decreased significantly, but values were significantly higher than those measured in fall. Males were spermiating from January to spawning in April.     

The influence of stocking density, light and temperature on the growth, production and nutrient removal capacity of Porphyra dioica (Bangiales, Rhodophyta)

The optimal conditions for growth of Porphyra dioica gametophytes were investigated in the laboratory, focusing on bioremediation potential. Porphyra dioica is one of the most common Porphyra species along the northern coast of Portugal and can be found year-round. The influence of stocking density and photon flux density (PFD) on the growth, production and nutrient removal was tested. Maximum growth rates, up to 33% per day, were recorded with 0.1 g fw l^− 1 at 150 and 250 μmol photons m^− 2 s^− 1. Growth rate decreased significantly with increasing stocking density. Productivity (g fw week^− 1) had an inverse trend, with more production at the higher stocking densities. At 150 μmol m^− 2 s^− 1 and with 1.5 g fw l^− 1, 1.4 g fw week^− 1 were produced. At this PFD, there was no significant difference in production between 0.6 to 1.5 g fw l^− 1. Nitrogen (N) content of the seaweeds decreased with increasing stocking densities and PFDs. The maximum N removal was recorded at 150 μmol m^− 2 s^− 1, with 1.5 g fw l^− 1 stocking density (1.67 mg N day^− 1). However, the N removed by thalli at 50 μmol photons m^− 2 s^− 1 was statistically equal to that at 150 and 250 μmol photons m^− 2 s^− 1, at a stocking density of 1.0 g fw l^− 1. The influence of temperature and photoperiod on growth and reproduction was also assessed. Growth rates of P. dioica were significantly affected by temperature and photoperiod. In this experiment (with 0.3 g fw l^− 1 stocking density), the highest growth rate, 27.5% fw day^− 1, was recorded at 15 °C and 16 : 8¯, L : D¯. Male thalli started to release spermatia 21 days after the beginning of the experiment, in temperatures from 10 to 20 °C and with 10, 12 and 16 h of day length. Unfertilized female-like thalli were observed at 10 to 20 °C, under all photoperiods tested. Growth of these thalli declined after 4 weeks. By then, formation of young bladelets in the basal portion of these thalli was observed. After 7 weeks all biomass produced was solely due to these vegetatively propagated young thalli, growing 22.4% to 26.1% day^− 1. The results of this study showed that P. dioica appears to be a candidate as a nutrient scrubber in integrated aquaculture systems.     

Effect of temperature and photoperiod on the growth, feed consumption, and biochemical content of juvenile green abalone, Haliotis fulgens, fed on a balanced diet

Juvenile green abalone, Haliotis fulgens (31.3 ± 0.1 mm, 3.7 ± 0.04 g live weight) were reared in laboratory for six months in order to determine their survival, growth, tissue composition, feed consumption (C), feed conversion ratio (FCR) and protein efficiency ratio (PER) under two temperatures (20 °C and 25 °C) and three photoperiods (00:24, 12:12 and 24:00 light:dark hours). Survival was ca. 100% at 20 °C, and between 68% and 75% at 25 °C. The highest gross growth rate (109 ± 4.3 μm d^− 1, 69 ± 3.9 mg d^− 1) was observed in abalone from the combination 20 °C-00:24 L:D. Slowest GGR (38 ± 4.1 μm d^− 1, 26 ± 1.5 mg d^− 1) was observed in the combination 25 °C-24:00 L:D. Organisms from 25 °C exhibited signs of the withering syndrome at the end of the experiment. Gross energy content (4.4 to 4.6 kcal g^− 1 tissue dry wt, TDW) and crude protein (60 to 68% TDW) were not significantly affected by either temperature or photoperiod, but organisms from 20 °C exhibited lower moisture content (86.8 to 88.6%) and higher tissue:shell ratio (0.34-0.40) than those from 25 °C (88.9 to 92.1%; 0.22-0.31 respectively). Feed consumption markedly increased at night, decreased with age, and was higher at 25 °C than 20 °C, irrespective of photoperiod. Highest C was observed under continuous darkness (0.66 to 0.95% BW d^− 1 at 20 °C and 0.84 to 1.25% BW d^− 1 at 25 °C), and was lowest under continuous light (0.50 to 0.82% BW d^− 1 at 20 °C and 0.71 to 1.02% BW d^− 1 at 25 °C). FCR and PER were both affected by temperature but not by photoperiod. Higher PER (2.2 to 3.4) and lower FCR values (0.69 to 1.05) were observed at 20 °C, when compared to 25 °C (PER 1.35 to 2.09, FCR 1.10 to 1.86). Sex ratios were ca. 1:1 in the 00:24 and 12:12 L:D photoperiods, yet ca. 50% of abalone from the 24:00 L:D photoperiod were immature at the end of the experiment. It is concluded that H. fulgens can be best cultured at 20 °C and 00:24 or 12:12 L:D regimes, while sustained temperatures at or above 25 °C may result in cumulative stress, altered physiological rhythms, and delayed maturation.     

Gill ATPase activities of silver perch, Bidyanus bidyanus (Mitchell), and golden perch, Macquaria ambigua (Richardson): Effects of environmental salt and ammonia

The Australian freshwater fish, silver and golden perch, are increasingly being used for aquaculture. Addition of salt to water is commonly used in commercial aquaculture to reduce stress attributed to high ammonia concentrations. The activities in gill homogenates of ouabain-sensitive Na⁺/K⁺-ATPase and NEM-sensitive ATPases (as a measure of H⁺-ATPases) of silver and golden perch were measured after maintaining the fish in water containing different salt and ammonia concentrations. Six treatments were applied in a 2 × 3 factorial design: two salt treatments, low salt (LS) of 2.5 g l^− 1 and high salt (HS) 5 g l^− 1, and three ammonia treatments, no added ammonia (NA), low ammonia (LA), 3 mg total ammonia nitrogen (TAN) l^− 1 and high ammonia (HA), 5 mg TAN l^− 1. In both species, activity of Na⁺/K⁺-ATPase was lowest in fish kept in the LSNA treatment (7.4 ± 0.4 μmol Pi mg protein^− 1 h^− 1 for silver perch and 3.1 ± 0.6 for golden perch) and highest in the HSHA treatment (15.2 ± 1.0 μmol Pi mg^− 1 protein h^− 1 for silver and 8.4 ± 1.2 for golden perch). In both species there was a significant increase (P < 0.001) in Na⁺/K⁺-ATPase activity with increase in salt concentration and with an increase in ammonia concentrations. A significant interaction (P < 0.036) between salt and ammonia on Na⁺/K⁺-ATPase activity was observed in silver but not in golden perch. In contrast, the lowest activity for NEM-sensitive ATPase was observed in the HSNA treatment (1.0 ± 0.2 μmol Pi mg^− 1 protein h^− 1 for silver and 1.5 ± 0.4 for golden perch) and highest in LSHA treatment (2.9 ± 0.4 μmol Pi mg^− 1 protein h^− 1 for silver and 3.6 ± 1.2 for golden perch). In both species there was a significant decrease in NEM-sensitive ATPase activity with increase in salt concentration and an increase in activity with increase in ammonia (P < 0.003). In silver perch, a significant interaction between the treatments was observed (P < 0.02). The results suggest that in these species of freshwater fish the Na⁺/K⁺-ATPase has a role in salt and ammonia homeostasis and that the NEM-sensitive ATPases are more active in fish kept in water with a lower salt content. It is possible that the increase in ammonia resistance when salt is added to the environmental water in commercial aquaculture systems may be due to the effects of salt on gill Na⁺/K⁺-ATPase activity rather than the NEM-sensitive ATPases.     

Routes of water loss in South African abalone (Haliotis midae) during aerial exposure

South African abalone, Haliotis midae, were exposed to air at 12 °C for 36 h to simulate the extent and rate mass loss experienced by animals during long distance live exports. Animals lost 15.1 ± 0.94% of their mass during the 36 h air exposure, an approximation of the highest mass losses sustained by industry.The total mass loss was attributed to water loss, as the contribution of dry mass to the total mass remained constant under all conditions. Water content decreased from 64.8% of the body mass (M_b) under control conditions to 58.8% M_b after 36 h in air. In real terms, however, animals had lost 22% of the body water pool.Abalone exhibited a typically high water turnover rate when in water (125 μL g^− 1 h^− 1), which decreased markedly during air exposure (2.2 μL g^− 1 h^− 1). Haemolymph volume decreased from 43% M_b in water to 14% M_b in air. The concomitant decrease in haemolymph pressure probably limited the first step in urine formation (ultra-filtration through the pericardium). Thus we observed that while urine flow represented about 26% of the total water loss when the animals were in water, urine flow ceased during air exposure.The decrease in haemolymph volume in air represents a redistribution of water to the tissues and not a bulk loss of haemolymph. This is supported by the concentration of haemolymph ions by a factor of 1.2 during aerial exposure, which was predicted based on the 22% decrease in water content. Under the same conditions, evaporation from water containers with similar surface to volume dimensions as abalone, accounted for only an 8.25% mass loss. As all other water loss routes were accounted for, we measured pedal mucus production rates of abalone in water and air. During 36 h aerial exposure, the pedal mucus production represented a loss of 6.8% M_b. We conclude that water loss during 36 h air exposure is attributable to evaporation (8.25% M_b) and pedal mucus production (6.8% M_b). This paves the way for directed research into mitigating water loss during the live export process.     

Growth, survival and biochemical composition of spider crab Maja brachydactyla (Balss, 1922) (Decapoda: Majidae) larvae reared under different stocking densities, prey:larva ratios and diets

The spider crab Maja brachydactyla is overexploited on the NW coast of Spain. Aquaculture of this species can be the solution to the problem, and consequently, several attempts of intensive larval rearing have been conducted. However, most of the studies already published do not provide enough zoo technical data, especially in terms of larval and prey densities or the nutritional quality of diets used for rearing.Three experiments were carried out to evaluate the conditions for intensive larval rearing of M. brachydactyla. Larval stocking density (10, 50 and 100 larvae L^− 1), prey:larva ratio (15, 30 and 60 enriched Artemia larva^− 1) and diet (enriched Artemia, non-enriched Artemia and polychaete supplement) effects on growth and survival of this species were studied. For larval culture nine, 35 L, 150 μm mesh-bottomed PVC cylinders (triplicates for each treatment and larval stage) connected to a recirculation unit, were used. Temperature and salinity were kept constant at 18 °C and 36‰ respectively. A 12 to 18 day trial was conducted for each experiment and samples of larvae were collected at each larval stage (zoea I, zoea II, megalopa) in the inter-molt phase and at first juvenile. Survival, carapace length and width, dry weight (DW), and proximate biochemical content (protein, carbohydrates and total lipid) as well as lipid class composition were determined.Stocking densities of 100 larvae L^− 1 resulted in higher growth in DW and higher content in lipids and protein for zoea I (ZI) and zoea II (ZII) than 10 larvae L^− 1. However, survival decreased with increasing stocking density.The use of 60 preys larva^− 1 produced larvae with significantly higher DW and protein content, especially at ZII stage, than lower prey densities. Survival rate obtained feeding 60 preys larva^− 1 up to the megalopa stage was almost two-fold (42.2%) the rate obtained using 15 preys larva^− 1 (24.8%).Larvae fed on enriched Artemia (EA) showed an increase in weight up to megalopa (518.9 ± 26.5 μg) in contrast to larvae fed on non-enriched prey (A) (467.9 ± 6.9 μg). Variation in DW correlated with the total lipid content (L) of the larvae (L_EA = 70.1 ± 37.5 μg ind^− 1; L_A = 28.9 ± 3.2 μg ind^− 1) especially in terms of neutral lipids. The use of an initial density of at least 50 larvae L^− 1 and 60 enriched Artemia larva^− 1 can be considered the most adequate rearing parameters in order to obtain good results in growth and survival of M. brachydactyla.     

An insulin-like system involved in the control of Pacific oyster Crassostrea gigas reproduction: hrIGF-1 effect on germinal cell proliferation and maturation associated with expression of an homologous insulin receptor-related receptor

The putative involvement of insulin-like peptides in the control of the reproduction of the Pacific oyster Crassostrea gigas was investigated using different approaches. In conjunction with a monthly histological analysis of the oyster reproductive cycle, in vitro biological effects of the human recombinant IGF-1 (hrIGF-1) on dissociated germinal cells were mesured over 1 year using [³H]-thymidine and [¹⁴C]-amino acid mixture as tracers for DNA and protein synthesis. DNA synthesis was stimulated by hrIGF-1 in November (114 ± 11% for 10^− 7M), December (46 ± 6% for 10^− 7 M) and January, which was identified as the highest gonial mitosis period. A clear dose-effect was observed in January with a maximum activation of 68 ± 7% for 10^− 12 M. Germinal cell protein synthesis was also stimulated in March (20 ± 1% for 10^− 10 M), April (41 ± 5% for 10^− 13 M), May (25 ± 4% for 10^− 13 M), and by almost all of hrIGF-1 doses in June (21.5 ± 2% for 10^− 13 M) and July (34 ± 1% for 10^− 13 M). This suggests the involvement of insulin-like substances in gonadal tubule rebuilding (December), as well as in the development of germinal cells (March, April), and in the summer maturation of gametes (May, June, July). These insulin-like effects conform with the expression pattern of the recently identified C. gigas insulin receptor-related receptor (CIR): It appeared highly expressed in the gonadal area during gonial mitosis phase, but also in maturating oocytes, suggesting the involvement of an insulin-like system in gonial proliferation and maturation. Moreover, CIR showed differential expression during embryogenesis and larval developmental stages. The expression of maternal CIR during the embryonic and early larval development, followed by the increasing zygotic CIR expression from D larvae to 11-day-old veliger larvae, then a decrease until metamorphosis, also suggest that insulin-like peptide is involved in organogenesis.     

Towards development of large-scale hatchery cultivation of larvae and spat of the pearl oyster Pinctada mazatlanica in Mexico

The results of a series of pilot-scale runs with P. mazatlanica larvae from 2004 through 2006 are reported. Preliminary runs in 2004 and 2005 used broodstock collected in summer, when massive spawning of wild populations naturally occurs. However, results of larval development were very poor and failed to produce spat in both years. In 2006, ripe broodstock were still collected in summer, but also in the spring time, based on the hypothesis that the gonads in this season were in better reproductive condition that in summer. Three larval runs were conducted in 2006: two in spring and one in summer. Larvae growth and survival greatly increased in both spring runs, ending with two successful productions of spat (∼ 20 × 10³ and ∼ 100 × 10³ juveniles). The summer larval run in 2006 failed again to produce spat. Additionally, the first run of April 2006 refers to an experiment that evaluated two different larval culture conditions: constant temperature (27 °C) and low stocking density (3-4 larvae ml^− 1) versus variable temperature (24-28 °C) and high stocking density (8-9 larvae ml^− 1). The first trial significantly increased larval survival and growth, which in turn resulted in greater numbers of settled spat, in comparison of the second trial, where survival, growth, and settlement of spat were significantly lower. Also in 2006, the quality of seawater used at the hatchery was evaluated with microbiological and chemical tests. The implication of these tests, together with results from all experiments are analyzed and discussed in terms of the potential development of large-scale hatchery cultivation of P. mazatlanica larvae in Mexico.     

Effects of dietary vitamin C on survival, growth, and immunity of large yellow croaker, Pseudosciaena crocea

An 8 weeks growth experiment was conducted to determine the effects of dietary vitamin C on the survival, growth, tissue ascorbic acid contents and immunity of large yellow croaker (Pseudosciaena crocea) with initial weight of 17.82 ± 0.68 g. Seven practical diets were formulated to contain 0.1, 12.2, 23.8, 47.6, 89.7, 188.5 and 489.0 mg ascorbic acid equivalent kg^− 1 diet, supplied as l-ascorbyl-2-polyphosphate (LAPP). Each diet was fed to triplicate groups of fish in seawater floating cages (1.5 × 1.5 × 2.0 m), and each cage was stocked with 120 fish. Fish were fed twice daily (05:00 and 17:00) to apparent satiation for 8 weeks. The water temperature fluctuated from 19.5 to 25.5 °C, the salinity from 25 to 28‰ and dissolved oxygen content was approximately 7 mg l^− 1 during the experimental period. Results showed that the specific growth rate (SGR) (from 1.80 to 1.96% d^− 1) had an increasing trend with the increase of dietary vitamin C, but no significant difference was observed among dietary treatments. No gross deficiency signs were observed in any of the experimental fish. Survival rate, however, significantly increased with increasing dietary vitamin C (P < 0.05). The vitamin C contents in liver and muscle correlated positively with the vitamin C in diets. The vitamin C requirement was estimated to be 28.2 mg kg^− 1 based on survival, and 87.0 mg kg^− 1 on liver content of vitamin C. The activities of serum lysozyme and alternative complement pathway (ACP), phagocytosis percentage (PP) and respiratory burst activity of head kidney significantly increased with increasing dietary vitamin C. The challenge experiment with Vibrio harveyi showed that fish fed the diets with supplementation of vitamin C had significantly lower cumulative mortality compared to the control group (66.7%), and the cumulative mortality (16.7%) in fish with 489.0 mg kg^− 1 ascorbic acid was significantly lower than that (41.7%) in fish with 23.8 mg kg^− 1 ascorbic acid. These results suggested that vitamin C significantly influenced the immune response and disease resistance of large yellow croaker.     

Effects of salinity and pH on ion-transport enzyme activities, survival and growth of Litopenaeus vannamei postlarvae

Effects of the salinity and pH on ion-transport enzyme activities, survival and growth of Litopenaeus vannamei postlarvae were investigated. Shrimp were transferred from salinity 31‰ and pH 8.1 to different salinity levels of 22, 25, 28, 31 (control), and to different pH levels of 7.1, 7.6, 8.1 (control), 8.6 and 9.1. The results showed ion-transport enzyme activities and weight gains of postlarvae were significantly affected by salinity and pH variation, which had no obvious effect on survival rate. The changing salinity affected the activities of total ATPase and Na⁺-K⁺-ATPase notably (F > F_0.05), meanwhile, non-significantly to the activities of V-ATPase, HCO₃⁻-ATPase. Within 48 h of salinity changing, the activities of ATPase, Na⁺-K⁺-ATPase in each treatment group gradually increased with the sampling time and reached their climax at 48 h, and then stabilized, showing negative correlation with salinity. The changing of pH affected greatly the activities of ATPase, Na⁺-K⁺-ATPase, V-ATPase and HCO₃⁻-ATPase (F > F_0.05), the activities of ATPase, Na⁺-K⁺-ATPase in each treatment group (pH = 7.1, 7.6, 8.6, 9.1) showed peak change within 72 h and stabilized afterwards, and the Na⁺-K⁺-ATPase activities came back to the level of the control group; Meanwhile the changing extent of V-ATPase and HCO₃⁻-ATPase activity corresponded with the grads of pH, and these ATPase activities showed negative correlation with pH changing, the activities of V-ATPase, HCO₃⁻-ATPase in postlarvae of each treatment group came to stable level after 24 h. The experiment also indicated the strength order of these ion-transport enzyme activities were as follows: Na⁺-K⁺-ATPase > V-ATPase > HCO₃⁻-ATPase. Na⁺-K⁺-ATPase was the chief undertaker of osmoregulation under the salinity effects, while V-ATPase and HCO₃⁻-ATPase were the chief osmoregulation undertakers under pH changing. In different salinity environment, the contributions of Na⁺-K⁺-ATPase, V-ATPase and HCO₃⁻-ATPase of L. vannamei postlarvae approximately accounted for 62.0-78.0%, 15.9-29.0% and 2.03-4.12% of ATPase activities in total, respectively. Meanwhile, in different pH medium, the contributions of these ATPases approximately accounted for 50.7-67.4%, 21.3-31.8% and 2.15-7.90% of total ATPase activities, respectively. Weight gain of shrimp transferred to salinity 31 (control) and 28‰ was significantly higher than that of shrimp reared at 25 and 22‰, and weight gain of shrimp transferred to pH 8.1 (control) and 8.6 was significantly higher that that of shrimp transferred to pH 7.1, 7.6 and 9.1. It was suggested that during the process of desalting and culturing of postlarvae, the salinity changing should not exceed 3 and pH variety not more than 0.5.     

Setting of a procedure for experimental incubation of Pacific oyster (Crassostrea gigas) embryos

Standardized experimental protocols designed to study Pacific oyster (Crassostrea gigas) gamete quality have not been previously published. Gamete sampling variations and confounding effect of interacting factors result in large variations between replicates, decreasing the effects of the studied factors. This work aims at defining a standardized procedure for incubation of oyster embryos designed for experimental purposes.In a first phase, four experiments were developed to improve embryo sampling and handling. They showed that a minimum of 50 embryos must be counted to decrease the variation between counting replicates. For reliable results, sampling must be carried out from 0 to 7 min after a careful agitation of seawater containing embryos (salinity: 35.6‰). Compared to control and a 30 cm one, the D-larval yield of embryos submitted to a 1 m fall was significantly reduced, showing the limited effect of mechanical disturbance on Pacific oyster oocytes survival. Then, maintaining 2.2 million embryos in 250 ml seawater for a two hours period resulted in a decrease of D-larval yield which could not be explained by a decrease of O₂ content and pH of seawater.The second phase of this work included a set of five experiments, defining experimental incubation conditions. A higher larval yield was observed using 1.8 l beakers and 150 l tanks compared to 0.3 ml microtiter plates and 1 l fish egg incubators. Higher larval yields were recorded when embryo density ranged from 5 to 100 ml^− 1, compared to values between 500 and 2000 ml^− 1. Compared to controls (no antibiotic or no presence of light), no changes of D-larval yield were observed by adding 20 ppm chloramphenicol or by maintaining embryos in total darkness. Then, a significant decrease of pH and O₂ content was observed during the incubation period. However, these changes could not be considered as limiting for Pacific oyster embryo development.In conclusion, experimental incubation conditions have been defined in this study: 30 embryos ml^− 1 incubated in 1.8 l beakers without antibiotic and regardless of light intensity, when not higher than 500 lux. The mean coefficients of variation observed between tank replicates ranged from 13.1 to 16.5%. The standardized incubation procedure described in this work will help to study quality variations of Pacific oyster gametes.     

The effects of temperature and photoperiod on ovarian development in captive grey mullet (Mugil cephalus L)

A series of sixteen environmental experiments, conducted under controlled laboratory conditions during the refractory period in the reproductive cycle of the grey mullet, determined the effect of photoperiod and temperature on the vitellogenesis of intraovarian oocytes. Fish subjected to the natural light cycle and ambient water temperatures (24–26°C) served as controls. A classification of stages of vitellogenesis (I–V) is used to determine the percentage composition of oocytes for each fish at intervals throughout the experiment following sampling in vivo.Onset of vitellogenesis is timed by the environmental conditions. A retarded photoperiod, irrespective of preconditioning photoperiod, plays a dominant role in stimulating oocyte growth. Temperature regulates vitellogenesis towards functional maturity. The combination of retarded photoperiod (6L/18D) and constant temperature of 21°C is the most effective for the completion of vitellogenesis of oocytes to functional maturity.Regular injections of pregnant mare's serum gonadotropin (PMSG) at 1 IU/g body weight are effective in initiating vitellogenesis.     

Growth,diet, and reproduction of Eurasian perch <Emphasis Type="Italic">Perca fluviatilis</Emphasis> L. in Lake Varese,northwestern Italy

No data have previously been reported on Eurasian perch Perca fluviatilis L. in Lake Varese. In this study, the growth, diet, and reproductive biology of the Eurasian perch population were investigated with the aim of providing information that may serve as a basis for efficient resource management. A total of 240 specimens were caught during the monthly sampling campaign from November 2006 through October 2008. The length-to-weight relationships were W _t = 8.4 × 10⁻³ L _t^3.10 (males) and W _t = 4.1 × 10⁻³ L _t^3.36 (females). The parameters for the von Bertalanffy growth function for pooled sexes were L _∞  = 33.17 cm, k = 0.20 year⁻¹, and t ₀ = −1.34 year. Perch in Lake Varese spawn from April through May. Sexual maturity is reached when males are 2 years old, in females mostly when they are 3 years old. Relative fecundity (F _rel) and absolute fecundity (F _abs) were assessed for females. Fecundity values were similar to data reported for other European populations: females of age 2+ F _rel = 102,457 ± 12,275, age 3+ F _rel = 131,767 ± 5,891, and age 4+ F _rel = 131,252 ± 15,555. Perch diet spectrum was wide and somewhat characterized by season. Perch in Lake Varese feed on macroinvertebrates, mainly Chironomidae and Chaoborus, zooplankton, and juvenile rudd Scardinius erythrophthalmus.     

Successful culture of larvae of Litopenaeus vannamei fed a microbound formulated diet exclusively from either stage PZ2 or M1 to PL1

In three separate experiments, harpaticoid copepods Tisbe monozota (alive and dead) and a microparticulate microbound diet were evaluated as alternatives to live Artemia nauplii as food, beginning at either stage PZ2 or M1, in the larval culture of Litopenaeus vannamei. Larvae were cultured in 2 L round bottom flasks at a density of 150 L^− 1 (Experiment 1) and 100 L^− 1 ( 3.2 and 3.3) at 28 °C, 35‰ salinity and 12:12 LD photoperiod, and fed 4×/day^- 1. Larvae were initially fed a mixture of phytoplankton to stages PZ2 or M1 and then fed either live Artemia, live or dead copepods, or a microparticulate microbound diet. The experiments were terminated and all larvae were harvested when more than 80% of larvae had molted to postlarvae 1 (PL1) within any flask representing any of the treatments. The comparative value of the different diets and feeding regimes was determined by mean survival, mean dry weight and total length of individual larva, and percentage of surviving larvae that were PL1. Trypsin activity of samples of larvae from each treatment was also determined. The microparticulate microbound diet effectively served as a complete substitute for Artemia nauplii when fed beginning at stage M1. When fed at the beginning of the PZ2 stage, survival was comparable to that of larvae fed Artemia, but mean dry weight, mean total length, and percent of surviving larvae that were PL1 generally were significantly less. Responses to the feeding of copepods, whether fed dead or live, as a substitute were generally significantly less than those of larvae fed either the Artemia nauplii or the microparticulate diet. Values of trypsin activity (10^− 5 IU/μg^- 1 dry weight) corresponded to the relative proportions of the different larval stages within a treatment, with higher activity being characteristic of early stages. Previously demonstrated successful results with another species of crustacean suggest that the microparticulate microbound diet has characteristics that should be effective in the culture of the carnivorous stages of other crustacean and fish larvae that are currently fed live Artemia nauplii.     

Effect of temperature on growth and energy budget of juvenile cobia (Rachycentron canadum)

Growth and energy budget of juvenile cobia (initial body weight ∼ 22 g) at various temperatures (23, 27, 31 and 35 °C) were investigated in this study. Maximal ration level (RL_max, %/day) increased as temperature (T, °C) increased from 23 °C to 31 °C but decreased at 35 °C, described as a quadratic equation: RL_max = −0.023T² + 1.495T − 17.52. Faecal production (f, mg g^− 1 day^− 1) increased with increased temperature (T, °C), described as a power function: lnf = 0.738lnT − 0.806. As temperature increased, feed absorption efficiency in dry weight (FAE_d, %), protein (FAE_p, %) and energy (FAE_e, %) all increased first and then decreased, but the variation of feed absorption efficiency was small, with ranges of 89.59-91.08%, 92.91-94.71%, 93.92-95.32%, respectively. Specific growth rate in wet weight (SGR_w, %/day), dry weight (SGR_d, %/day), protein (SGR_p, %/day) and energy (SGR_e, %/day) showed a domed curve relative to temperature (T, °C), described as quadratic equations: SGR_w = − 0.068T² + 3.878T − 50.53, SGR_d = − 0.079T² + 4.536T −59.64, SGR_p = − 0.084T² + 4.783T − 63.08 and SGR_e = − 0.082T² + 4.654T − 60.99, and SGR_w, SGR_d, SGR_p and SGR_e maximized at 28.5 °C, 28.6 °C, 28.4 °C, 28.5 °C, respectively, as calculated from the regression equations. The relationships between feed conversion efficiency in wet weight (FCE_w, %), dry weight (FCE_d, %), protein (FCE_p, %), energy (FCE_e, %) and temperature (T, °C) also took on a domed curve described as quadratic equations: FCE_w = − 0.726T² + 39.71T − 473.8, FCE_d = − 0.276T² + 15.31T − 190.6, FCE_p = − 0.397T² + 22.05T − 277.9 and FCE_e = − 0.350T² + 19.39T − 239.9, and FCE_w, FCE_d, FCE_p and FCE_e maximized at 27.4 °C, 27.8 °C, 27.7 °C and 27.7 °C, respectively, as calculated from the regression equations. Energy budget of juvenile cobia fed satiation was 100C = 5F + 67(U + R) + 28G at water temperature 27 °C and 100C = 5F + 70(U + R) + 25G at water temperature 31 °C, where C is food energy, F is faeces energy, (U + R) is excretion energy and metabolism energy, and G is growth energy.