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 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.     

Production of astaxanthin from Haematococcus in open pond by two-stage growth one-step process

We have developed a two-stage growth one-step process for cultivation of Haematococcus using a self-designed system that mimics an open pond in the natural environment. The characteristics of this process are green vegetative cell growth and cysts transformation and pigment accumulation that proceed spontaneously and successively in one open photobioreactor. Four strains of Haematococcus (H. pluvialis 26; H. pluvialis 30; H. pluvialis 34; H. pluvialis WZ) were cultured in this imitation system for a duration of 12 days. The changes in cell density and medium pH were closely monitored, and the astaxanthin content and yield of the four Haematococcus strains were measured at the end of 12 days of cultivation. Two of the strains, H. pluvialis 26 and H. pluvialis WZ, were selected as strains suitable for mass culture, resulting in the astaxanthin yield of 51.06 and 40.25 mg L^− 1 which are equivalent to 2.79 and 2.50% of their dry biomass respectively. Based on the laboratory work, 6 batch cultures of H. pluvialis WZ were conducted successfully to produce astaxanthin in two 100 m² open race-way pond by two-stage growth one-step process. The astaxanthin content ranged from 1.61 to 2.48 g 100 g^− 1dry wt., with average astaxanthin content of 2.10 g 100 g^− 1dry wt. Compared with the one-stage production of astaxanthin based on continuous culture, the superiority of our process is that it can accumulate much more astaxanthin in red cysts. Compared with two-stage production of astaxanthin, the advantage of our process is that it does not need to divide the production process into two parts using two bioreactors. The presented work demonstrates the feasibility for producing astaxanthin from Haematococcus using a two-stage growth one-step process in open pond, culture systems that have been successfully used for Spirulina and Chlorella mass culture. The future of Haematococcus astaxanthin production has been also discussed.     

The effects of algal diets on population growth and egg hatching success of the tropical calanoid copepod, Acartia sinjiensis

As natural diets of fish larvae, a number of calanoid copepod species are being investigated for use as live prey in aquaculture hatcheries. One of these, the tropical calanoid copepod, Acartia sinjiensis, has good potential as a live feed for tropical reef fish larvae. However, the rearing techniques for A. sinjiensis require further development to improve productivity. This study was carried out to investigate the population growth and egg hatching success of A. sinjiensis when fed a range of mono-and binary algal diets, including algae in the form of frozen paste.For the population growth experiment, the final A. sinjiensis population, including eggs, nauplii, copepodites and adults, was determined after feeding eight algal diets (two frozen algae, four live monoalgal and two live binary algal diets) for 8 days at temperature 28 ± 1 °C; salinity 34 ± 1 psu and photoperiod 12 L:12 D. Five replicates, with an initial 12 adult A. sinjiensis per replicate, were set up for each treatment. In a separate experiment, effects of diets on egg hatching success were examined after 48 h incubation of eggs produced by A. sinjiensis fed the same eight diets.The results showed that diet significantly affected both population growth and hatching success of A. sinjiensis. Of the diets tested, the binary algal diets were more successful than monoalgal diets, while the frozen algae had little dietary value. The highest population growth was recorded on A. sinjiensis fed a binary diet of Tetraselmis chuii and the Tahitian strain of Isochrysis sp. (T-ISO) (final population: 1091 ± 80), which was significantly higher (P < 0.001) than all other diets tested except for the other binary diet of Nannochloropsis sp. and T-ISO (final population: 897 ± 123). Diet also had a significant effect (P < 0.001) on egg hatching rate, though the highest hatch rate was recorded with eggs produced by A. sinjiensis fed binary diet Nannochloropsis sp. and T-ISO (88.1 ± 2.1%), this was not significantly different from that of eggs produced by A. sinjiensis fed either T-ISO alone (88.0 ± 1.7%) or the binary algal diet of T. chuii and T-ISO (76.4 ± 7.1%). The results of this study suggest that among the diets tested, a combination of live T. chuii and T-ISO was the best for the culture of A. sinjiensis.     

Growth and gonadal development in diploid and triploid turbot (Scophthalmus maximus)

This study determined the effect of triploidy on the survival, growth and gonadal development of turbot from 6 to 48 months of age. From 6 to 24 months of age (first sexual maturity), survival was similar in both ploidies (P > 0.05). From 24 to 48 months of age, after the first sexual maturity, survival was 91.9% in diploids and 100% in triploids, which did not exhibit the post-spawning-associated mortality. Growth was similar for both ploidies during the first year of life. After that, triploids grew significantly (P < 0.05) more that diploids, with more marked differences after each spawning season. From 24 to 48 months, the average weight difference between both ploidies was 11.4 ± 1.9%, ranging from 4.3 to 23.0%. At 47 months of age, the biomass of triploids was 10.3% greater in total weight and 14.3% greater in eviscerated weight. Gonads of triploid males were similar to that of diploids, whereas in triploid females, they were significantly smaller and rudimentary. A histological analysis carried out at 47 months of age showed complete sterility of triploids in both sexes. Sex ratio was 1 male (M):0.6 female (F), for diploids, significantly (P < 0.05) different from 1:1, and 1 M:3.3 F for triploids, significantly (P < 0.05) different from 1:1 and from the diploids. Since females grow more than males, culture of triploids benefited from the high female ratio, which helped to reduce size dispersion. In addition, their sterility allowed better performance by avoiding the reduction in growth that takes place during the spawning periods. Together, these observations indicate that triploidy induction can be an interesting option for turbot aquaculture, especially for the production of large-size fish of more than 2 years of age.     

Waste excretion of marble goby (Oxyeleotris marmorata Bleeker) fed with different diets

Marble goby (Oxyeleotris marmorata Bleeker), with its high demand and price, has a great potential as a profitable commercial aquaculture candidate in Malaysia and Southeast Asia region. Efforts are being made to produce this species in a better controlled culture environment like recirculating aquaculture system (RAS) due to poor growth performance and disease problems shown by conventional cage and outdoor pond culture systems. Quantification of waste excreted by fish is critical to RAS design. This study was conducted to characterize the waste excretion rates of marble goby fed with different diets (live food and minced fish). Ammonia-N (TAN), urea-N, nitrite-N (NO₂-N), nitrate-N (NO₃-N), total-N (TN), organic-N (ON), feces-N, 5-day biochemical oxygen demand (BOD₅) and total suspended solid (TSS) produced from marble goby were determined over a 72-h excretion period. Under given experimental conditions, the results showed that feed type had significant influence on the waste excretion rates, with marble goby fed live tilapia (Oreochromis niloticus) exhibiting significantly (P < 0.05) the lowest amount of waste excretion comparable to that of fish fed live common carp (Cyprinus carpio) and minced scads (Decapterus russellii). This indicates that feeding marble goby with tilapia poses less adverse effects on water quality and is thus a suitable diet for this species. The waste excreted by the fish is composed of nitrogenous excretion (TAN, Urea-N, ON, Feces-N), and productions of dissolved biodegradable organic substances (BOD₅) and TSS (TSS_feces + TSS_water). About 58-71% of the nitrogen consumed in food was excreted and its rate depended mainly on the feed type. TAN was the chief end-product of protein metabolism; about 74-84% of the daily total nitrogenous excretion was TAN. Urea-N accounted for 13-21% of the daily total nitrogenous excretion indicating that urea-N is an important nitrogenous excretory end-product in marble goby. The waste excretion data presented in this study can be served as a pre-requisite for designing a RAS for this species. The overall BOD₅ and TSS production found in this study also point to the need for including bio-filtration unit and suspended solids removal mechanism in the RAS design.     

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.     

Astaxanthin digestibility as affected by ration levels for Atlantic salmon, Salmo salar

The objective of this study was to determine the effect of feed intake on the apparent digestibility coefficient (ADC) of astaxanthin including the major geometrical E/Z-isomers in Atlantic salmon (Salmo salar). Atlantic salmon (50 per pen, initial weight 2 kg) were kept in 125 m³ sea pens equipped with an excess feed collection system to monitor and quantify accurate feed intake, and subjected to three treatments in triplicate. All salmon were fed the same diet supplemented with 47 mg astaxanthin per kg. Two treatments were fed full or restricted rations corresponding to 100 or 40% of apparent satiation, respectively, with a switch in ration between two consecutive feeding periods (14 and 3 d, respectively), whereas the control treatment was fed to 100% of apparent satiation during both periods. The corresponding feed intakes were 0.45 and 0.16% of biomass for salmon fed 100% or 40% of apparent satiation, respectively. Faeces were collected by stripping at the end of each feeding period and ADCs of astaxanthin were determined by an indirect method using yttrium oxide (Y₂O₃) as an indigestible marker. Feed intake and astaxanthin ADC were negatively correlated (R² = 0.64; p = 0.0001). Astaxanthin ADC was 1.5 times higher at the low compared to the high ration level (p < 0.05), but due to the low feed intake the total amount of digested astaxanthin was only about 50% of that in fish fed to satiation. The ADCs of the all-E and 13Z-isomers of astaxanthin were similar and considerably higher than for 9Z-astaxanthin (p < 0.05). The amount of digested astaxanthin/TGC (estimated thermal growth coefficient) decreased with increasing feed intake. Estimates of astaxanthin retention indicated that a higher feed intake cause a lower muscle concentration of astaxanthin compared to that obtained at low feed intakes due to the lower digestibility. Blood samples were collected at the end of the second feeding period to examine possible relationships between plasma carotenoids, feed intake and ADC. Salmon without faeces in the hindgut had a lower plasma astaxanthin concentration compared to salmon with faeces in the hindgut (p < 0.05). In conclusion, feed intake and astaxanthin ADC are negatively correlated, and may thus explain reductions in muscle retention of Atlantic salmon.     

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.     

Metazoan parasites on gills of Southern Bluefin Tuna (Thunnus maccoyii) do not rapidly proliferate after transfer to sea cages

A relatively new and highly valuable aquaculture industry focuses on three species of bluefin tunas, which are captured from the wild and fattened for several months in sea cages. In teleost aquaculture, mortalities and extra production costs are very commonly associated with metazoan ectoparasites. In tuna, however, the production value lost due to diseases associated with ectoparasites is unknown. We collected epidemiological data on burdens of metazoans on the gills of farmed southern bluefin, Thunnus maccoyii, in a series of monthly samples of tuna from the time of stocking through to harvest (March to August, 2004; N = 210) in five sea cages on a farm off Port Lincoln, Australia. Three species were recorded; for one (a copepod, Pseudocycnus appendiculatus), there was a gradual, significant increase in both abundance (from a mean of 0.1 in March, to 3.83 in August) and prevalence (from 10% to 67.5%). For the other two species (a second copepod, Euryphorus brachypterus, and a polyopisthocotylean flatworm, Hexostoma thynni) there were no discernible trends in prevalences and abundances. These results contrast markedly with those of other intensively cultured species of finfishes, in which parasite epizootics are frequent. This finding may indicate that despite the stresses of captivity, tuna mount a robust immune response to ectoparasites; the relatively low stocking densities at which tuna are farmed may facilitate this. The fall in water temperature during farming (22 °C to 13 °C) may also reduce the reproductive rate of these ectoparasites.     

Prey ingestion and live food selectivity of marble goby (Oxyeleotris marmorata) using rice field prawn (Macrobrachium lanchesteri) as prey

Marble goby (Oxyeleotris marmorata), a carnivorous fish native to freshwater in Asia-Pacific region, is a high-valued species in many Asian countries. The present study consisting of three experiments was conducted to determine the appropriate density, size and ingestion time of marble goby fingerlings on rice field prawn (Macrobrachium lanchesteri) as prey. Results showed that the ingestion rate of marble goby fingerlings (0.26-1.43 g size) on prawns (9-12 mm in length) had an asymptotic pattern peaked at a predator to prey ratio of 1:20. Marble goby fingerlings did catch prawns during both daytime (07:00-17:00 h) and nighttime (17:00-07:00 h), but the ingestion peaked during nighttime period. The hourly ingestion rate during nighttime was significantly higher than that during daytime for the smaller-size marble goby fingerlings of 0.26-0.87 g, while there was no significant difference in the hourly ingestion between nighttime and daytime for the larger-size marble goby fingerlings of 1.43 g. The daily ingestion of marble goby fingerlings on prawns (mg consumed prawns/goby/day) increased significantly with increasing average weights (g) of marble goby fingerlings (Y = 27.6 + 47.8X, n = 30, r² = 0.86, p < 0.05). Results also showed that the preferred prawn size of marble goby fingerlings of average weights of 0.26-4.18 g was small (9-12 mm) with positive electivity index of 0.19-0.33, and the shift from small- to medium-size (12-14 mm) prawns was slow. The present study has demonstrated the feasibility of using rice field prawns as live foods to nurse marble goby fingerlings.     

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 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.     

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 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.     

Development of phytoplankton communities and common off-flavors in a biofloc technology system used for the culture of channel catfish (Ictalurus punctatus)

The use of biofloc technology production systems continues to increase in the aquaculture industry worldwide. Recent research demonstrated that outdoor biofloc systems can be used to produce high yields of channel catfish (Ictalurus punctatus). However, studies have not yet been performed to determine the development and composition of phytoplankton communities and related off-flavor problems in these biofloc production systems. In this study, water samples were collected biweekly from May to November and channel catfish samples were collected during harvest in November from nine 18.6 m² biofloc culture tanks. Water and fillet samples were analyzed for levels of the common off-flavor compounds geosmin and 2-methylisoborneol (MIB). The development and composition of phytoplankton communities in each culture tank was also monitored. In addition, water and biofloc samples were evaluated to assess the microbial sources of geosmin and MIB within the culture tanks. Phytoplankton (including algae and cyanobacteria attached to bioflocs) biomass, as determined by concentrations of chlorophyll a in the water, gradually increased in all tanks over time. Phytoplankton communities that developed in the culture tanks were dominated by fast-growing, unicellular and small colonial types of green algae (chlorophytes) and diatoms (bacillariophytes) and slower growing, small colonial types of cyanobacteria (cyanophytes). A positive correlation (p < 0.05) between cumulative feed addition and chlorophyll a concentration was found. Although geosmin and MIB were present in the culture water of each tank during most of the study, levels were typically low and only one tank yielded catfish with geosmin and MIB in their flesh at levels high enough to be designated as off-flavor. A positive correlation (p < 0.05) between cumulative feed addition and MIB concentrations in the water of culture tanks indicates a greater potential for MIB-related off-flavor problems at high feed application rates. The microbial sources responsible for production of geosmin and MIB in the culture tanks remain unknown.     

Influence of dietary soybean meal levels on growth, feed utilization and gut histology of Egyptian sole (Solea aegyptiaca) juveniles

The present study was undertaken in order to determine the effect of a dietary incorporation of soybean meal (SBM) on growth performance, nutrient utilization and gut histology of Egyptian sole Solea aegyptiaca. This species, which is genetically and morphologically close to Solea solea, is currently present in the Mediterranean Sea and reared in some Italian farms, using the same techniques for S. solea production and it could represent a promising new species for Mediterranean aquaculture.Three isoproteic and isolipidic extruded diets (crude protein, 47%, crude fat, 20%) were formulated containing 0 (diet 1), 18 (diet 2) and 30% SBM (diet 3). 540 juveniles (initial body weight of 6.9 ± 0.2 g) were equally distributed into nine 150 l squared tanks (bottom surface: 520 cm²) connected with a recirculation system (temperature 20 ± 1 °C and dissolved oxygen above 6 ppm). Animals have been hand-fed twice a day (at 9.00 a.m. and 5.00 p.m.) at a daily ratio of 1.5% body weight^− 1. Diets were tested in triplicate for 87 days. The fish were weighed at days 28, 57 and 87, and they were sampled at days 0, 57 and 87 for nutrient retention determination. One-way ANOVA with Newman-Keuls' post-test were used to analyze data (P ≤ 0.05).No differences in terms of palatability were observed and any diet has been consumed completely all throughout the experiment. At day 87, fish fed diet 1, 2 and 3 reached weights of 25.8 ± 0.8 g, 27.5 ± 0.9 g and 26.1 ± 1.5 g, respectively and no differences in performances or nutrient retention were found.Histopathological gut examinations have revealed no noticeable differences in the appearances of the intestines between any of diet groups. Intestinal mucosal cells presented well vacuolated upper, with well defined microvillar brush borders. On the basis of the results of this trial, SBM seems to be a good protein source for Egyptian sole and can be added in the diet up to 30% without any reduction in growth rate and no adverse effect on gut histology.     

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.     

Phaffia rhodozyma as an astaxanthin source in salmonid diets

The red yeast Phaffia rhodozyma has possible application as a component of diets for use in aquaculture. Its primary value lies in its content of astaxanthin, which is much higher (5–50 times) than that found in crustacean meals. When fed to rainbow trout, the deposition of astaxanthin in the fish flesh was dependent on the proper preparation of yeast cells before their inclusion into the feed. No astaxanthin was nutritionally available from intact yeast. If P. rhodozyma was mechanically ruptured its pigments were transferred to the flesh of rainbow trout, coloring it salmon-pink. The most efficient deposition of astaxanthin in trout occurred when the cell wall of P. rhodozyma was partially removed by enzymatic digestion.The proxiamte composition, amino acid content, fatty acid profile, and astaxanthin content were determined for P. rhodozyma. When grown under the conditions used in this study, P. rhodozyma has a low protein content (～25%) and a high total lipid content (～17%) compared with most other microorganisms. Its amino acid profile is well balanced but is deficient in methionine. The predominant fatty acids present in the yeast are oleic, linoleic and palmitic acids.     

Hydroponic horticulture using residual waters from Oreochromis niloticus aquaculture with biofloc technology in photoautotrophic conditions with Chlorella microalgae

Protein‐dependent aquaculture generates large amounts of nutrient‐rich residuals; a feasible way to develop sustainable production systems is to integrate Decoupled Aquaponic Systems (DAPS) with residual water bioprocesses, to combine Photoautotrophic Biofloc Technology (P‐BFT) aquaculture and hydroponic horticulture. This study describes the characteristics of residual water from Oreochromis niloticus aquaculture performed with P‐BFT inoculated with Chlorella microalgae, reared during the nursery (180 fish m³) and grow‐out (55 fish m³) phases. The experiment included five treatments: photoautotrophic BFT inoculated with Chlorella sp. (M), C. sorokiniana 2714 (CV), and C. sorokiniana 2805 (CS), and chemoautotrophic (Q) and heterotrophic (H) as controls. Elemental characteristics in liquid and solid residual fractions (15 macro‐ and micronutrients) were compared among treatments and against Hoagland & Arnon solution with hydroponics and used in Nutrient Film Technique (NFT) hydroponic horticulture including five plant species: lettuce (Lactuca sativa), pak‐choi (Brassica rapa subsp. chinensis), rocket (Eruca sativa), spinach (Spinacia oleracea) and basil (Ocimum basilicum). The physicochemical parameters were ideal for O. niloticus and plants. The relationship between N:P was ideal until weeks 16–22 in the photoautotrophic treatments, compared with hydroponic solutions. Micronutrient content was greater in the solid than a liquid fraction. The best BFT effluent regarding fish and plant growth was photoautotrophic treatments. Oreochromis niloticus BFT aquaculture in photoautotrophic mode using microalgae Chlorella inoculations provided residual water beneficial to hydroponic horticulture in DAPS located in coastal arid zones where freshwater is scarce, improving aquaculture performance and reusing water and nutrients.