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.     

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%.     

Productivity and photosynthetic efficiency of outdoor cultures of Tetraselmis suecica in annular columns

In this study, 120-l annular columns were used to cultivate Tetraselmis suecica outdoors. The mass transfer at different aeration rates and the influence of the harvest rate on productivity and biochemical composition were investigated. The potential of the system was evaluated by estimating productivity at full-scale. Two different arrangements to simulate a full-scale plant and determine the “overall areal productivity” (OAP) were experimented with. In August 2003, one experimental column (full-scale column) was placed between seven dummy columns. All the reactors were positioned at a distance of 0.8 m wall to wall and centred at the vertices of equilateral triangles. A second experimental column (isolated column) was placed in a separate area under full sunlight. In August 2004, the columns were placed side by side in an east-west oriented row at a distance of 0.24 m wall to wall.In the first experiment, the mean volumetric productivity of the full-scale column was not significantly lower than that achieved by the isolated column (0.46 against 0.49 g l^− 1 day^− 1) in spite of the shading by the dummy units. The average OAP and efficiency of conversion of visible solar radiation (PE) were 36.3 g m^− 2 day^− 1 and 9.4%, respectively. In the second experiment, the full-scale column attained a mean volumetric productivity of 0.42 g l^− 1 day^− 1. The OAP and the PE were 38.2 g m^− 2 day^− 1 and 9.3%, respectively.     

Density dependent ambient ammonium as the key factor for optimization of stocking density of common carp in small holding tanks

Advanced fry of common carp (1.6 ± 0.2 g) were reared in experimental outdoor tanks (4500 l; 3 × 1.5 × 1 m) for 312 or 151 days under six stocking conditions of 8, 13, 16, 32, 48 and 64 fish per tank for ascertaining the threshold and critical levels of ammonium and, hence, to recommend the optimum stocking density of common carp for culture under rearing stage conditions. The samples of water were monitored from each tank at regular intervals for water quality parameters as well as for ammonium concentrations. Fishes were harvested at the end of the experiment. The results revealed a significant decrease in fish growth as stocking density increased. Absence of mortality and favorable growth resulted in maximum fish biomass at the stocking density of 16 fish/tank, but the heavy mortality and stunted growth caused the poor total fish biomass in the highest stocking density employed. The interactions between ammonium and fish growth were expressed at three different concentration levels of ammonium: (a) favorable concentration range (0.262-0.294 mg l^− 1), (b) growth inhibiting concentration range (0.313-0.322 mg l^− 1) and (c) lethal concentration range (0.323-0.422 mg l^− 1). The ambient ammonium concentrations of 0.313 mg l^− 1 (or equivalent ammonia concentration of 0.0342 mg l^− 1) and 0.323 mg l^− 1 (or equivalent ammonia concentration of 0.043 mg l^− 1), observed for stocking density ranging from 17 to 19 fish per tank, were considered to be the threshold and critical levels of ammonium that caused growth inhibition and mortality of fish. Fish mortality was higher when the ratio of DO to ammonium remained quite low (< 15), but no mortality occurred with higher ratio. Considering the economic viability of the production system, this appears that the optimum fish stocking density would be around SD 16 (equivalent to 210 g m^− 3).     

Intensive production of juvenile tiger shrimp Penaeus monodon: An evaluation of stocking density and artificial substrates

Tiger shrimp Penaeus monodon were intensively grown from PL₁₅ for 56 d in tank systems at stocking densities of 1000 and 2000 shrimp m^− 3, with and without the addition of artificial substrates (AquaMat® (buoyant and non-buoyant) and polyethylene mesh) at each density. Shrimp growth was significantly greater at the lower density and when substrates were added. Mean shrimp weight at harvest ranged from 0.64 ± 0.06 g (2000 shrimp m^− 3, no added substrate) to 1.17 ± 0.01 g (1000 shrimp m^− 3, added substrate). Survival was high and averaged 79.5 ± 2.7% across all treatments. The addition of substrates significantly increased survival at both stocking densities; however, survival was not significantly affected by stocking density. A maximum harvest density of 1645 shrimp m^− 3 and biomass of 1.27 kg m^− 3 were produced at a stocking density of 2000 m^− 3 with added substrates. Both harvest density and biomass significantly increased with stocking density and addition of substrates. The feed conversion ratio (FCR) of formulated feed was significantly lower when substrates were added. The results show that growth of P. monodon juveniles was inversely related to stocking density during intensive production. However, production output was significantly increased by addition of artificial substrates, which enhanced both growth and survival.     

Exploring Northeast American and Asian species of Porphyra for use in an integrated finfish-algal aquaculture system

Many aquaculture industries generate a nutrient-rich waste stream that can lead to eutrophication of coastal waters. To address this environmental issue, the bioremediation potential of several native Northeast American species of Porphyra was assessed and compared to the well-known Asian species. Porphyra thalli were cultured over 4 weeks at 15 °C at a stocking density of 0.4 g FW L^− 1. At 3- to 4-day intervals nutrient uptake, tissue N accumulation and phycobiliprotein concentration (PBP) were determined as functions of nitrogen (N) concentration (25-300 μM) and N source (nitrate vs. ammonium). Growth rates were measured weekly. Growth and tissue N reached maximal levels at inorganic N concentrations of 150-300 μM. Maximum growth rates ranged from 10% to 25% day^− 1, although induction of archeospores reduced average growth rates in many cases. No evidence of ammonium toxicity (reductions in growth rate) was observed; in fact, similar values were found with both N sources. Ammonium generally yielded higher PBP and tissue N contents than nitrate. Porphyra amplissima presented the highest growth rate, followed by the Asian Porphyra yezoensis. Under the experimental conditions, Porphyra spp. removed 70-100% of N within 3-4 days at N concentrations up to 150 μM, but was less efficient in removing inorganic phosphorus (35-91% removal). The highest tissue N and PBP concentrations were found at 150-300 μM of N, with N values close to 7% DW. Overall, Porphyra appears to be an excellent choice for bioremediation of moderately eutrophic effluents, with the added benefit that tissue may be harvested for sale.     

Rearing of African catfish (Clarias gariepinus) and vundu catfish (Heterobranchus longifilis) in traditional fish ponds (whedos): Effect of stocking density on growth, production and body composition

African catfish (Clarias gariepinus) (initial body weight: 34.8 ± 4.8 g) and vundu catfish (Heterobranchus longifilis) (initial body weight: 39.1 ± 8.2 g) fingerlings were stocked at densities of 4, 6 or 8 fish m^− 3 in traditional fish ponds (whedos) constructed in the floodplain of the Oueme River (South Benin, West Africa), for 70 days from March to June 2005. Fish were fed twice a day with 34% crude protein feed formulated with locally available ingredients. The effects of stocking density were evaluated in growth responses, gross production and body composition. Water quality variables were similar (p > 0.05) in all compartments. Temperature and pH were at the optimum level for fish. Dissolved oxygen ranged from 0.9 to 1.2 mg l^− 1 during the experiment and secchi disc transparency was low (< 14 cm). In both species, growth responses increased with the increasing density, significantly in African catfish stocked at density of 8 fish m^− 3 compared to the other densities (4 and 6 fish m^− 3) but not significantly in vundu catfish. Production data ranged from 3.1 ± 0.5 to 22.8 ± 4.5 t ha^− 1 year^− 1 in African catfish and from 6.1 ± 1.2 to 15.1 ± 3.1 t ha^− 1 year^− 1 in vundu catfish. Production increased with increasing stocking densities but only significantly (p < 0.05) between the density of 8 fish m^− 3 and the other densities. In both species, carcass fat increased with increasing density (p < 0.05) while carcass protein and moisture decreased (p > 0.05). These results are important because they indicate that, as far as growth rate and production are concerned, African catfish is more profitable than vundu catfish for culture at high density in whedo.     

The effects of stocking density on gonad growth, survival and feed intake of adult green sea urchin (Strongylocentrotus droebachiensis)

In this study the effects of different stocking densities on survival, gonad growth and feed intake in adult green sea urchin (Strongylocentrotus droebachiensis) were examined. Two size groups of sea urchins with respective initial mean weight of 45 g (S) and 70 g (L) were used. The experiments were carried out in 755 l plastic tanks divided into chambers by vertical fibreglass lamellae. In the two experiments stocking densities of 6, 12, 14 and 16 kg m^− 2 (Exp I) and 3, 6 and 8 kg m^− 2 (Exp II) were used. Sea urchins reared individually in compartmentalised raceways were used as controls in both experiments. All groups were fed a formulated moist feed ad libitum for 60 days at 8 °C (± 0.5). Mortality was observed in all groups, except for the control group and the lowest density group of large sea urchins (3 kg m^− 2, L). Both mortality and occurrence of injuries increased significantly with increasing stocking density in both experiments and for both size groups. In the highest density groups mortality was 60% (L) and 80% (S). There was a significant increase in gonad index during the experimental period in both experiments. Increased stocking density had a significantly negative effect on gonad growth in both size groups while feed intake was unaffected. The results show that adult S. droebachiensis can maintain high survival rate and high gonad growth at stocking densities up 6 kg m^− 2 in holding facilities of the design used in the present study.     

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.     

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.     

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.     

Studies on the biofiltration capacity of Gracilariopsis longissima: From microscale to macroscale

The potential of the red agarophyte Gracilariopsis longissima as biofilter for phosphate and ammonium in effluents outflowing intensive marine fish cultures was assessed at different scales. Previous studies showed that both laboratory (microscale level) and outdoor cultivation (mesoscale level) were feasible, with a maximum sustainable yield of 270 g fresh wt m^− 2 day^− 1 approximately, at a biomass higher than that predicted in a logistic model, a deviation attributable to an improvement of the culture conditions during the monitoring period. At a mesoscale level, a 34-h cycle suggested that the nitrification rate on the seaweed fronds showed diel fluctuations, with rates peaking early in the morning, when ammonium uptake rates were negligible. Mean nitrification rates were similar to ammonium uptake rates, suggesting that nitrifyers outcompete G. longissima for the use of ammonium; especially when mean biofiltering efficiencies were less than 15% during the 34-h period.G. longissima thrives naturally in different earthen ponds of a fish farm in Cádiz Bay Natural Park, Southern Spain, especially in the outflowing reservoir earthen ponds, where biomass reached values up to 278 g dry wt m^− 2 during the spring. A field cultivation system for G. longissima (macroscale level) was designed to find the best scenario in terms of earthen pond, season or current conditions. The best cultivation method was the growth of vegetative cuttings on suspended braided nylon ropes. The highest growth rates (up to 6% day^− 1) and biomass (up to 10 g fresh wt cm^− 1 rope) were obtained in ponds receiving outflow waters, suggesting a nutrient effect. The net P production reached 24.9 μg P cm^− 1 rope day⁻¹ and was also higher on braided nylon suspended ropes placed at the outflowing reservoir earthen ponds. A similar result was found regarding net N production. However, in this case, mean production (≈ 170 μg N cm^− 1 rope day^− 1) was similar in the different earthen ponds and channels. The increase in P and N biomass suggested that G. longissima was biofiltering efficiently nutrient wastes from the fish farm.The results pointed out the high potential ability of G. longissima to biofilter waste waters from a fish farm, encouraging a large scale cultivation of this species. Future practices using this macroalgae may be implemented in local fish farms, resulting in both environmental and economic advantages.     

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.     

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.     

The effect of stocking density of Chinese mitten crab Eriocheir sinensis on rice and crab seed yields in rice-crab culture systems

Normally Chinese mitten crab Eriocheir sinensis were cultured for 2 years, 1 year is cultured from crab larvae to seeds or button sized crab in ponds, tanks or rice fields, the another year is cultured from seeds to market size in ponds, reservoirs or other waters. The present study was conducted to determine the effect of stocking density (0, 3.75, 15, 30 and 60 inds. per m², respectively) of Chinese mitten crab larvae on rice and crab seed yields in rice-crab culture systems using land-based enclosures (4 m × 7 m, with three replicates for each stocking density) at the Panjin Guanghe Fisheries Co. Ltd, in the Liaohe Delta, China. Zoobenthos biomass, production of crab and rice paddy, and economical profit among the treatments were determined. The experiment showed that zoobenthos biomass in rice-crab culture systems was relatively variant and not significantly correlated to these stocking density. The biomass of aquatic plant declined significantly with increasing stocking density. Specific growth rates (SGR) and survival rates of the crab were significantly higher at the lower stocking density (P < 0.05); in addition, the ratio of precocious crab and total crab significantly increased with the stocking density increasing (P < 0.05); There were not significant effect of presence of crab on yields of rice straw and rice paddy (P > 0.05). Net crab yield and net profit were the highest at the 15 treatment (P < 0.05), however, the net profits among 3.75, 15 and 30 treatments did not show significant difference (P > 0.05) economically. Based on the observations above, a stocking density of 3.75 to 30 inds. per m² was considered reasonable in rice-crab culture system.     

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.     

Vegetative propagation of the carrageenophytic red alga Gigartina skottsbergii Setchell et Gardner: Indoor and field experiments

The carrageenophytic red alga Gigartina skottsbergii presents several biological constraints for its cultivation such as restricted temporal availability and high spore mortalities that affect the development of its mass cultivation. For this reason, research to develop alternative propagation methods has been undertaken. Previous laboratory studies demonstrated that manipulating temperature, light and nutrients could enhance healing and regeneration of this seaweed. In this study nursery and field experiments were undertaken to establish the possibility to regrow G. skottsbergii in conditions similar to those applied in mass cultivation practices. Frond fragments and rhizoids were tested as alternative ways to obtain new plants. Our results indicate that regeneration occurs in the field, and can be managed in nursery conditions. The addition of a nitrogen source enhances the growth from 0.6 to 1.0% d^− 1 of the healed fragments and the use of a photon flux density above 50 μmol m^− 2 s^− 1 increases the growth rate but decrease the survival of the explants. Explants can be transferred to field conditions and grow at similar rates to those registered in the nursery (0.5% d^− 1). It is also possible to propagate rhizoids of G. skottsbergii and it seems that the attachment of a frond portion to the substratum, enhanced its survival and regeneration capacity. Finally, this study demonstrated that rhizoids attached to rocks can regenerate complete plants in nature, a feature which could be a useful for developing a sustainable harvesting methodology.     

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.     

Growout production of camouflage grouper, Epinephelus polyphekadion (Bleeker), in a tank culture system

Growout production of the camouflage grouper, Epinephelus polyphekadion (Bleeker), in a 10-m³-capacity fibreglass tank culture system was evaluated, using hatchery-produced fingerlings (56-59 g initial weight) at stocking densities of five, 15 and 45 fish m^?3. During the first 9 months of a 12-month growout period, the fish were fed twice a day with a moist pellet feed containing 40.9% protein. From month 10 onwards until harvest, the fish were fed moist pellets in the morning and trash fish in the evening at a 1:1 ratio. The final weight of fish at harvest was up to 900 g, with mean weights of 544.6 ± 170.72 g at five fish m^?3, 540.2 ± 150.82 g at 15 fish m-^?3 and 513.3 ± 134.52 g at 45 fish m^?3. The results showed no significant differences (P > 0.05) in growth rate and fish size between the different stocking densities tested. The average daily growth rate ranged from 0.62 to 3.38 g fish^?1 day^?1, with mean weights of 1.49 ± 0.74 g fish^?1 day^?1 at five fish m^?3 through 0.53 to 2.38 g fish^?1 day^?1, 1.32 ± 0.57 g fish^?1 day^?1 at 15 fish m^?3 to 0.48-3.32 g fish^?1 day^?1 and 1.31 g fish^?1 day^?1 at 45 fish m^?3 stocking density. Although up to 100% survival was observed at the lowest stocking density, the survival rate significantly decreased (P < 0.05) with increasing stocking density. The food conversion ratio (FCR) significantly decreased (P <0.05) with increasing stocking densities, showing efficient feed utilization with increasing stocking densities of E. polyphekadion. The FCR averaged 2.1 at a stocking density of 45 fish m^?3. The yield in terms of kg fish produced m^?3 of water used in the culture system significantly increased (P < 0.001) from five to 45 fish m^?3. The yield averaged 17.3 ±0.53 kg m^?3 at a stocking density of 45 fish m^?3. The present results show that the present tank culture system could sustain more biomass in terms of increasing fish stocking densities. The growth performance of E. polyphekadion observed during this investigation has been reviewed with other grouper species.