Effects of Salinity on Growth and Energy Budget of Juvenile Cobia,Rachycentron canadum

The effects of salinity on the growth and energy budget of juvenile cobia, Rachycentron canadum, were evaluated. Triplicate tanks with ten fish per tank (initial weight 17.58 ± 0.26 g/fish, mean ± SD) reared at salinities of 5, 10, 15, 20, 25, 30, and 35 ppt were fed with fresh squid to satiety for 15 d. Results indicated that there were no significant differences in daily ration level in wet weight (RL_w), dry weight (RL_d), and energy (RL_e) of the fish. There were also no significant variations in daily fecal production (f_e) and apparent digestibility coefficient of energy (ADC_e) among salinity treatments. Specific growth rates (SGRs) in wet weight (SGR_w), dry weight (SGR_d), and energy (SGR_e) showed domed curves relative to salinity. Quadratic regression analyses of SGR_w, SGR_d, and SGR_e against salinity indicated that the optimal salinity for maximal growth of juvenile cobia was 29.9, 29.9, and 28.5 ppt, respectively. Similar to the trend of SGR, food conversion efficiency for juvenile cobia in wet weight (FCE_w), dry weight (FCE_d), and energy (FCE_e) increased with the increases in salinity, maximized at 30 ppt, and then decreased when salinity reached 35 ppt.     

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 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.     

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.     

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.     

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.     

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.     

The effects of light and temperature on the photosynthesis of the Asparagopsis armata tetrasporophyte (Falkenbergia rufolanosa), cultivated in tanks

The integrated aquaculture of the tetrasporophyte of Asparagopsis armata Harvey (Falkenbergia rufolanosa) using fish farm effluents may be viable due to the species high capacity of removing nutrients and its content of halogenated organic compounds with applications on the pharmaceutical and chemical industries. In order to optimize the integrated aquaculture of F. rufolanosa, we followed the daily variation of the potential quantum yield (F_v/F_m) of PSII on plants cultivated at different biomass densities and different total ammonia nitrogen (TAN) fluxes to check if they are photoinhibited at any time of the day. Moreover, the photoinhibition under continuous exposure to highly saturating irradiance and its potential for subsequent recovery in the shade was assessed. The potential for year round cultivation was evaluated by measuring rates of O₂ evolution of plants acclimated at temperatures ranging from 15 to 29 °C, the temperature range of a fish farm effluent in southern Portugal where an integrated aquaculture system of F. rufolanosa was constructed.Photoinhibition does not seem to be a major constrain for the integrated aquaculture of F. rufolanosa. Only when cultivated at a very low density of 1.5 g fresh weight (FW) l^− 1 that there was a midday decrease in maximal quantum yield (F_v/F_m). At densities higher than 4 g FW l^− 1, no photoinhibition was observed. When exposed to full solar irradiance for 1 h, F. rufolanosa showed a 33% decrease in F_v/F_m, recovering to 86% of the initial value after 2 h in the shade. A midday decline of the F. rufolanosa F_v/F_m was also observed under the lowest TAN flux tested (∼6 μM h^− 1), suggesting that this fast and easy measurement of fluorescence may be used as a convenient diagnostic tool to detect nutrient-starved unbalance conditions of the cultures. Maximum net photosynthesis peaked at 15 °C with 9.7 mg O₂ g dry weight (DW)^− 1 h^− 1 and remained high until 24 °C. At 29 °C, the net oxygen production was significantly reduced due to a dramatic increase of respiration, suggesting this to be the species' lethal temperature threshold.Results indicate that F. rufolanosa has a considerable photosynthetic plasticity and confirm it as a good candidate for integrated aquaculture at temperatures up to 24° C and cultivation densities of at least 5 g FW l^− 1. When cultivated at these densities, light does not penetrate below the first few centimetres of the surface zone. Plants circulate within the tanks, spending around 10% of the time in the first few centimetres where they are able to use efficiently the saturating light levels without damaging their photosynthetic apparatus.     

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.     

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.     

Determinism of the induction of the reproductive cycle in female Eurasian perch, Perca fluviatilis: Identification of environmental cues and permissive factors

In Eurasian perch, Perca fluviatilis, gametogenesis is induced by decrease of both temperature and photoperiod. However, a multiplicity of other factors can display non-permissive or modulating effects on the induction of the reproductive cycle. Consequently, a 9-week study was carried out to determine the environmental cues and permissive factors controlling the induction of the reproductive cycle in this species. A two-level fractional factorial design 2^8-4 was used in order to identify the influent factors from 8 environmental and nutritional ones and their 28 interactions. Tested factors were: (1) amplitude of temperature decrease, (2) time, (3) kinetics and (4) amplitude of photoperiod decrease, (5) initial nutritional state, (6) handling, (7) feeding rate and (8) light spectrum. Increase of gonadosomatic index (GSIi), proportion of female in exogenous vitellogenesis (EVP), plasma 17 β-estradiol (E₂) and cortisol levels, fat consumption and food intake were evaluated.This study showed that the amplitude of the decrease of temperature and the time of photoperiod decrease are the two main environmental cues controlling the induction of the reproductive cycle in Eurasian perch. GSIi was the highest (3.8%) when females were exposed to both low amplitude of temperature decrease and precocious decrease of photophase. It was the lowest (1.1%) when high amplitude of temperature decrease and late decrease of photophase were applied. Handling in association with temperature modulated the broodstock response. 100% of the unhandled fish held at 14 °C were at the exogenous vitellogenesis stage with mean E₂ and basal cortisol levels of 1.6 ng mL^− 1 and 9 ng mL^− 1 respectively. Handled fish and fish held at 6 °C exhibited lower vitellogenesis (40-73%) associated with lower E₂ (0.6-1.1 ng mL^− 1) and higher basal cortisol levels (37-89 ng mL^− 1). No other factor nor interaction displayed a cueing or non permissive effect on the induction of the reproductive cycle in female Eurasian perch. A first schematic model is proposed to explain the factorial determinism of the induction of the reproductive cycle.     

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.     

Experimental integrated aquaculture of fish and red seaweeds in Northern Portugal

Three potentially valuable red seaweeds, Chondrus crispus Stackhouse, Gracilaria bursa pastoris (S.G. Gmelin) P.C. Silva and Palmaria palmata (L.) O. Kuntze, collected in northern Portugal, were cultivated using the nutrient-rich effluents from a local turbot (Scophthalmus maximus Linnaeus) and sea bass (Dicentrarchus labrax Linnaeus) farm. The algae were cultivated in a two level cascade system. Several arrangements of the cascade system, stocking densities (3, 5, 7 and 8 kg m^− 2) and water fluxes (140 and 325 l h^− 1) were tested to optimize biomass yield and nitrogen uptake rate and efficiency. The yield and the total ammonium nitrogen (TAN) uptake of the three species were highly seasonal. Palmaria could not survive culture conditions during the summer when water temperature was above 21 °C. In the spring, Palmaria had an average yield of 40.2 (± 12.80) g DW m^− 2 day^− 1 and a nitrogen uptake efficiency (NUE) of 41.0% (± 17.26%). NUE expresses, in percentage, the average reduction in TAN concentration between the inflows and the outflows of the tanks. Chondrus performed better in summer with an average yield of 37.0 (± 11.10) g DW m^− 2 day^− 1 and removing 41.3% (± 17.32%) of nitrogen. Gracilaria grew year round, but also performed better during spring/summer, producing an average of 29.1 (± 2.90) g DW m^− 2 day^− 1, and only 7.3 (± 5.08) g DW m^− 2 day^− 1 during autumn. Yield of C. crispus did not differ significantly when grown at two different stocking densities (5 kg m^− 2 and 8 kg m^− 2). On the other hand, Gracilaria had significantly higher yields at 5 than at 7 kg m^− 2. Better NUE, on average 76.7% (± 22.13%), was also obtained with 5 kg m^− 2 stocking density and only 63.8% (± 24.62%) with 7 kg m^− 2. The yield of Gracilaria increased significantly with the increase of water flux from 140 to 325 l h^− 1 and more nitrogen was removed from the water. However, NUE decreased from 48.4% to 33.4% at 140 and 325 l h^− 1, respectively. Biofiltration was highly improved by a cascade system with a NUE as high as 83.5%.     

The tetrasporophyte of Asparagopsis armata as a novel seaweed biofilter

The red seaweed Asparagopsis armata (Harvey; Rhodophytae, Bonnemaisoniaceae) produces biologically active secondary metabolites that are valuable natural ingredients for cosmetics and medicine and its cultivation may therefore be a profitable venture. The tetrasporophyte of this species (“Falkenbergia rufolanosa”) was successfully tank-cultivated as a continuous biofilter for the effluent of a commercial fish farm in southern Portugal. Optimal stocking density for highest biomass yield and a low level of other algal species in winter and late spring was 5×g centrifuged fresh weight l^− 1. The effect of total ammonia nitrogen supply (TAN flux) on biofiltration and biomass yield was investigated in winter and spring. Results revealed that A. armata is currently the seaweed-biofilter with the highest TAN removal of up to 90 μmol l^− 1 h^− 1 at a TAN flux of about 500 μmol l^− 1 h^− 1. In the tanks used, this is equivalent to a removal of up to 14.5 g TAN m^− 2 day^− 1. At a lower TAN flux of about 40 μmol l^− 1 h^− 1, TAN removal by A. armata is more than double to what is reported at this flux for another successful seaweed biofilter, the genus Ulva. Monthly variation of A. armata biomass yield peaked in May and was lowest in January. At TAN fluxes between 300 and 400 μmol l^− 1 h^− 1, an average water temperature of 21.7 °C and a total daily photon flux density of 47 Mol m^− 2, seaweed yield was over 100 g DW m^− 2 day^− 1 with a recorded maximum of 119 g. During spring, autumn and early summer, the biomass of A. armata within the experimental tanks doubled every week. A model for the up scaling of this finfish integrated aquaculture of A. armata varies the investment in biofilter surface area and estimates the return in biofiltration and biomass yield. Highest TAN removal efficiencies will only be possible at low TAN fluxes and a very large biofilter area, resulting in a low production of biomass per unit area. To remove 50% of TAN from the effluent (1 mt Sparus aurata; 21 °C), 28 m² of biofilter, designed to support a water turnover rate of 0.8 Vol h^− 1 would be necessary. This system produces 6.1 kg FW (1.5 kg DW) of A. armata per day and has the potential to turn biofiltration into an economically sustained, beneficial side effect.     

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.     

Distribution of a fish pathogen Listonella anguillarum in the Japanese flounder Paralichthys olivaceus hatchery

The present study was undertaken to investigate the distribution of Listonella anguillarum in a Japanese flounder (Paralichthys olivaceus) hatchery. A total of 2704 isolates were obtained from the developing fish, live diets and artificial feeds of Japanese flounder and their rearing water, 439 of which were identified as L. anguillarum by the combining incubation on thiosulfate-citrate-bile salt-sucrose (TCBS) agar at 35 °C overnight with polymerase chain reaction (PCR) detection for the VAH1 hemolysin gene. L. anguillarum was detected in all seven rotifer samples, with densities of 2.5 × 10³ to 4.6 × 10⁶ colony forming units (CFU) g^− 1. Both the analyzed samples of Nannochloropsis oculata contained this bacterium at densities of 1.6 × 10⁴ to 1.4 × 10⁵ CFU g^− 1. L. anguillarum was detected in only one of four samples of Artemia nauplii with a density of 4.8 × 10⁵ CFU g^− 1 (35%) and it was not detected in the two analyzed artificial feed samples. L. anguillarum was detected in 11 of 18 specimens of larval and juvenile Japanese flounder at densities of 5.0 × 10¹ to 7.4 × 10⁵ CFU g^− 1, while it was not detected in the two analyzed egg specimens of Japanese flounder. These results indicate that L. anguillarum associated with the developing Japanese flounder is likely derived from rearing water and live diets such as rotifers. Further, it is strongly suggested that L. anguillarum is a transient bacterium of the intestinal microflora for the Japanese flounder but is a permanently indigenous one for the Japanese flounder hatcheries.     

First feeding of Octopus vulgaris Cuvier, 1797 paralarvae using Artemia: Effect of prey size, prey density and feeding frequency

Different assays related to the first feeding of Octopus vulgaris Cuvier, 1797 are compiled in this paper. They include: age at initial feeding age, prey size selection and optimal density, attack timing after feeding, and effect of dose number on the number of captures. Prey capture and ingestion processes were also analysed. Food supplied was cultured Artemia sp. Each assay lasted 15 min.Although paralarvae already start to feed on the hatching day (day 0), it is during day 2 when a greater number of attacks is recorded (81.7 ± 14.7% paralarvae attack). They mainly prefer (significance level α = 0.05) large Artemia, 1.4 ± 0.4 mm (77.0 ± 5.6% of the total attacks) than small Artemia, 0.8 ± 0.1 mm (23.0 ± 5.6%). There is also a slight predilection for the lowest Artemia concentration (33.3 ±12.6% paralarvae attack in a 0.1 Artemia ml^− 1 density, opposite 16.7 ± 7.6 and 18.3 ± 7.6% in densities of 0.5 and 1 Artemia ml^− 1 respectively). The greatest predatory activity is recorded during the first 5 min after food is supplied (72.2 ± 25.5%). An increase in the predatory activity was also observed when food was distributed in several doses instead of a single dose (75.0 ± 10.0% and 46.7 ± 17.6% respectively). It was proved for the first time that paralarvae completely ingest their preys (including their exoskeletons), in this case Artemia. Time needed for their total ingestion ranges between 4 and 10 min.     

Sedimentation from mussel (Perna canaliculus) culture in the Firth of Thames, New Zealand: Impacts on sediment oxygen and nutrient fluxes

Shellfish aquaculture is growing worldwide and previous studies have shown a range of associated environmental impacts. In the Firth of Thames, New Zealand, there are more than 2000 ha of existing and approved farm area with applications pending for approximately another 6000 ha, however, no previous studies have examined the impacts of mussel culture in this region. To determine the impact of a mussel farm (45 ha) in the Firth of Thames we measured sedimentation rates by deploying sediment traps, sediment characteristics by collecting sediment cores and sediment oxygen and nutrient fluxes by deploying benthic chambers in four seasons. Sedimentation under the farm was increased by 106 g m^− 2 d^− 1 compared to the reference site. Similarly sediments under the farm had elevated organic carbon, nitrogen, chlorophyll a and phaeopigment concentrations indicative of the additional organic input due to bivalve biodeposition. Oxygen consumption was higher under the farm compared to a reference site (1.1-2.1×) but this increase was only significant (p < 0.001) in summer when rates reached 3083 μmol m^− 2 h^− 1 under the farm. Ammonia release rates ranged from 80 to 319 μmol m^− 2 h^− 1 and were higher under the farm compared to the reference site in spring (1.8×, non-significant p = 0.588) and autumn (3×, significant p = 0.006) but in summer release rates at the reference site (275 μmol m^− 2 h^− 1) were 1.4× higher than those under the farm. Nitrate fluxes (3.1 to 21.8 μmol m^− 2 h^− 1) were significantly (p = 0.001) higher at the farm site. Oxygen and nutrient fluxes generally demonstrated the typical response to increased organic input due to sedimentation from mussel culture. Unusually low nitrogen release rates in summer may indicate enhanced denitrification under the farm. Benthic regeneration at the reference site could supply 74% of nitrogen required by pelagic primary producers whereas under the farm it could account for 94%. This demonstrates the importance of benthic nutrient regeneration in this region and that mussel culture can lead to a redistribution of nutrients. The farm we studied is small and located in a high-energy environment and impacts from larger farms or in areas where biodeposit dispersal is limited are likely to be even more significant and we suggest that site-specific hydrodynamic and biogeochemical conditions have to be taken into account when planning new mussel farms to prevent excessive modifications of nutrient dynamics.     

The effects of fish hydrolysate (CPSP) level on Octopus maya (Voss and Solis) diet: Digestive enzyme activity, blood metabolites, and energy balance

As has been demonstrated in previous studies, Octopus maya can be fed on artificial diets. In the present study six different diets were assayed. Five diets were designed to test the effect of percentage of inclusion of fish protein concentrate (CPSP: 0, 5, 10, 15, and 20%) and were offered to octopuses as a specifically designed artificial diet. The sixth diet consisted of frozen crab (Callinectes spp) and was used as control diet. Blood metabolites and energy budget of octopuses were evaluated to determine how CPSP levels modulate the digestive capacity and allow retaining energy for growth. Wild animals (316.4 ± 9.8 g) were used in the study. Results showed that CPSP produced a positive specific growth rate (SGR, % day^− 1) with high value in octopuses fed 15% CPSP level. A maximum growth rate of 0.86% day^− 1 was recorded in these animals, a value that is extremely low when compared with the SGR obtained when animals were fed fresh crab (3.7% day^− 1). In general, blood metabolites were affected by diet composition, indicating that some metabolites could reflect the nutritional and/or physiological status of octopus. Preliminary reference values for O. maya fed crab were found for glucose (0.09 ±0.02 mg/ml), lactate (0.004 ± 0.002 mg/ml), cholesterol (0.16 ± 0.02 mg/ml), acylglycerol (0.14 ± 0.01 mg/ml), protein (0.37 ± 0.04 mg/ml), hemocyanin (1.85 ± 0.04 mmol/l), and digestive gland glycogen (1.86 ± 0.3 mg/g). Total energy content can be used as an indicator of tissue metabolic reserves. In the present study, higher energy content in the digestive gland and muscle was observed in octopuses fed crab, followed by animals fed 15% CPSP. Results from the digestive gland indicated that the retained energy derived from glycogen, suggesting that lipids and protein were the main sources of variation linked with energy content. In general, digestive gland proteases activity and trypsin were induced in octopuses fed 15% CPSP. The capacity of O. maya juveniles to adjust their digestive enzymes to different types of food was evidenced. Essential amino acid content (EAA) of the diet was not a limiting factor. When dietary EAA profiles were compared with O. maya EAA profiles, all dietary EAA resulted in a higher concentration than whole body octopus composition. In the present study, all experimental groups ingested between 3300 and 4106 kJ wk^− 1 kg^− 1 without statistical differences among treatments, indicating that experimental diets were as attractive as crab. Differences were recorded in the proportion of absorbed energy (Ab, %) between CPSP-based and crab meat diets, suggesting digestion limitations associated with artificial diets. The present results indicate that the 15% CPSP diet had characteristics that stimulate digestive enzymes and reduce energetic costs associated with its digestion (HiE or SDA), channeling more biomass production than the other experimental diets.     

Selective particle feeding by the Chilean oyster, Ostrea chilensis; implications for nursery culture and broodstock conditioning

Hatchery broodstock conditioning and nursery culture of the Chilean flat oyster Ostrea chilensis have been hampered by the poor performance of oysters fed typical microalgal hatchery diets. To determine the feeding capabilities of this species the selective removal and consumption of natural planktonic assemblages and artificial inert particles (polystyrene beads) by juvenile and adult oysters were examined experimentally. The arrangement of the eulaterofrontal cirri of the ctenidia was also examined to infer their potential efficiency of particle selection for feeding. Polystyrene beads of 45 and 15 μm in diameter had high rates of removal from suspensions by both juvenile (45 μm = 70%, 15 μm = 73%) and adult (45 μm = 88%, 15 μm = 87%) oysters. In contrast, beads of 6 μm diameter had lower rates of removal (adults = 68%, juveniles = 53%), while 1 μm beads were not removed at all. Both adult and juvenile oysters feeding upon natural plankton assemblages removed only microphytoplankton (20-75 μm in size) despite the presence of nanophytoplankton (2-20 μm), picophytoplankton (< 2 μm), cyanobacterium Synechoccocus spp. (< 2 μm), and bacterial cells (< 75 μm) in the experimental suspensions. Eulaterofrontal cirri of both juvenile (15.2 μm ± 0.9 SE) and adult oysters (18.9 μm ± 0.3 SE) are the shortest that have been reported for any ostreid species which helps to explain the inability of this species to retain small food particles. The clearance rates for oysters feeding on microphytoplankton only were 1.49 (± 0.05 SE) and 7.1 (± 1.2 SE) l h^− 1g^− 1 for juveniles and adults respectively. These values are much higher than previously reported for this species being fed smaller sized cultured microalgae. Our results strongly suggest that the difficulties in the nursery and broodstock hatchery culture of this oyster may be due to inappropriate phytoplankton diets. We recommend the provision of cultured microalgae of 20-75 μm in diameter for improving the performance of hatchery maintained juvenile and adult O. chilensis.