Constructed wetlands as a treatment method for effluents from intensive trout farms

This study examined the effects of different hydraulic loading rates on the treatment efficiency of subsurface flow (SSF) constructed wetlands treating effluents from trout farming over a period of 6 months. Six identical wetland cells with a pre-sedimentation zone of 9.6 m² and a root zone of 23.6 m² were loaded with effluents from intensive trout farming (> 2.1 kg feeding stuff per L/s and day). The total runoff of 13.2 L/s was treated in the wetland cells, where two duplicate cells received equal hydraulic loads of 3.9, 1.8 and 0.9 L/s. All examined wetland cells had significant treatment effects on the nutrient fractions containing particulate matter [total nitrogen (TN), total phosphorous (TP), biological oxygen demand in 5 days (BOD₅), chemical oxygen demand (COD), and total suspended solids (TSS)].

Efficiency was between 5.5% for TN and 90.1% for TSS. The SSF wetland also had a high treatment effect on total ammonia nitrogen (TAN), with efficiencies of 61.2 to 87.8%. Nitrate nitrogen (NO₃–N) and phosphate phosphorous (PO₄–P) showed a significant increase in the wetland effluent by 8.4 to 209%. Nitrite nitrogen (NO₂–N), had no significant, or significant effluent increase depending on the inflow rate. Treatment efficiency for particulate nutrients and TAN increased with decreasing hydraulic load, while the differences between 1.8 and 0.9 L/s were not significant. The treatment efficiency for TP was constant for all cells, at around 40%. The wetland receiving 3.9 L/s was over-flooded after 10 to 12 weeks due to colmatation. Nevertheless, the wetland still showed high treatment efficiencies. For commercial trout farms, SSF wetlands are a highly effective method of effluent treatment. A hydraulic load of 1 L/s on 13.3 m² wetland area (1.8 L/s on the examined wetland) seems most suitable. Higher loads lead to accelerated wetland colmatation, while lower loads waste space.     

Wood chips and wheat straw as alternative biofilter media for denitrification reactors treating aquaculture and other wastewaters with high nitrate concentrations

This study evaluated wood chips and wheat straw as inexpensive and readily available alternatives to more expensive plastic media for denitrification processes in treating aquaculture wastewaters or other high nitrate waters. Nine 3.8-L laboratory scale reactors (40 cm packed height × 10 cm diameter) were used to compare the performance of wood chips, wheat straw, and Kaldnes plastic media in the removal of nitrate from synthetic aquaculture wastewater. These upflow bioreactors were loaded at a constant flow rate and three influent NO₃–N concentrations of 50, 120, and 200 mg/L each for at least 4 weeks, in sequence. These experiments showed that both wood chips and wheat straw produced comparable denitrification rates to the Kaldnes plastic media. As much as 99% of nitrate was removed from the wastewater of 200 mg NO₃–N/L influent concentration. Pseudo-steady state denitrification rates for 200 mg NO₃–N/L influent concentrations averaged (1360 ± 40) g N/(m³ d) for wood chips, (1360 ± 80) g N/(m³ d) for wheat straw, and (1330 ± 70) g N/(m³ d) for Kaldnes media. These values were not the maximum potential of the reactors as nitrate profiles up through the reactors indicated that nitrate reductions in the lower half of the reactors were more than double the averages for the whole reactor. COD consumption per unit of NO₃–N removed was highest with the Kaldnes media (3.41–3.95) compared to wood chips (3.34–3.64) and wheat straw (3.26–3.46). Effluent ammonia concentrations were near zero while nitrites were around 2.0 mg NO₂–N/L for all reactor types and loading rates. During the denitrification process, alkalinity and pH increased while the oxidation–reduction potential decreased with nitrate removal.

Wood chips and wheat straw lost 16.2% and 37.7% of their masses, respectively, during the 140-day experiment. There were signs of physical degradation that included discoloration and structural transformation. The carbon to nitrogen ratio of the media also decreased. Both wood chips and wheat straw can be used as filter media for biological denitrification, but time limitations for the life of both materials must be considered.     

Dopamine modulates the physiological response of the tiger shrimp Penaeus monodon

Levels of glucose, lactate, pO₂, pCO₂, HCO₃⁻, TCO₂, Na⁺, K⁺, Cl⁻, protein, and oxyhemocyanin in the hemolymph and its osmolality and pH were measured when tiger shrimp, Penaeus monodon (13.5 ± 1.5 g body weight), were individually injected with saline or dopamine at 10^− 8, 10^− 7, or 10^− 6 mol shrimp^− 1. Results showed that hemolymph glucose, lactate, pCO₂, HCO₃⁻, and TCO₂ values increased from 2 to 4 h; hemolymph osmolality, Na⁺, and total protein had increased at 2 h; and hemolymph K⁺ decreased from 2 to 8 h after the dopamine injection. All physiological parameters returned to the control values 4–16 h after receiving dopamine. The dopamine injection also significantly decreased the oxyhemocyanin/protein ratio of P. monodon which occurred at 2 h, resulting from an elevation of hemolymph protein and a slight decrease of oxyhemocyanin. These results suggest that stress-inducing dopamine caused a transient period of modulation of energy metabolism, osmoregulation, respiration, and the acid–base balance in P. monodon in adapting to this environmental stress.     

Treatment of flow-through trout aquaculture effluents in a constructed wetland

A study on effluent treatment with sub-surface flow (SF) constructed wetlands was conducted in a small commercial scale Bavarian (Germany) flow-through trout farm. Under limited spatial and financial conditions a most suitable wetland was constructed. The wetland treatment efficiency at high hydraulic loading rates during raceway runoff and cleaning situation in comparison to sedimentation as initial treatment method was examined.

The constructional solution involved the alteration of six existing sedimentation basins (SB) to SF horizontal flow constructed wetlands with a pre-sedimentation area. As constructional materials only local, cheaply available materials were used in order to reduce the costs. The SF wetland had high treatment efficiencies in the two operational modes examined. During cleaning situation at a hydraulic loading rate (HLR) of 13.6 m/day treatment efficiency for total suspended solids (TSS) was highest and reached 68%. While during raceway runoff situation total ammonia nitrogen (TAN) treatment efficiency of 88% overtopped the efficiency of the other nutrients examined at a HLR of 10.6 m/day. In both treatment situations the SF wetland efficiency was significantly higher than the effect of the SB. SF constructed wetlands treating high hydraulic loading rates accompanied with short retention times were effective on dissolved nutrient treatment only for TAN and nitrite nitrogen (NO₂–N), while other dissolved nutrients like nitrate nitrogen (NO₃–N) and phosphate phosphorous (PO₄–P) showed no or even negative treatment effects through the wetland passage. To reduce these nutrients, other treatment conditions or wetland configurations are needed.     

Comparing denitrification rates and carbon sources in commercial scale upflow denitrification biological filters in aquaculture

Aerobic biological filtration systems employing nitrifying bacteria to remediate excess ammonia and nitrite concentrations are common components of recirculating aquaculture systems (RAS). However, significant water exchange may still be necessary to reduce nitrate concentrations to acceptable levels unless denitrification systems are included in the RAS design. This study evaluated the design of a full scale denitrification reactor in a commercial culture RAS application. Four carbon sources were evaluated including methanol, acetic acid, molasses and Cerelose™, a hydrolyzed starch, to determine their applicability under commercial culture conditions and to determine if any of these carbon sources encouraged the production of two common “off-flavor” compounds, 2-methyisoborneol (MIB) or geosmin. The denitrification design consisted of a 1.89 m³ covered conical bottom polyethylene tank containing 1.0 m³ media through which water up-flowed at a rate of 10 lpm. A commercial aquaculture system housing 6 metric tonnes of Siberian sturgeon was used to generate nitrate through nitrification in a moving bed biological filter. All four carbon sources were able to effectively reduce nitrate to near zero concentrations from influent concentrations ranging from 11 to 57 mg/l NO₃–N, and the maximum daily denitrification rate was 670–680 g nitrogen removed/m³ media/day, regardless of the carbon source. Although nitrite production was not a problem once the reactors achieved a constant effluent nitrate, ammonia production was a significant problem for units fed molasses and to a less extent Cerelose™. Maximum measured ammonia concentrations in the reactor effluents for methanol, vinegar, Cerelose™ and molasses were 1.62 ± 0.10, 2.83 ± 0.17, 4.55 ± 0.45 and 5.25 ± 1.26 mg/l NH₃–N, respectively. Turbidity production was significantly increased in reactors fed molasses and to a less extent Cerelose™. Concentrations of geosmin and MIB were not significantly increased in any of the denitrification reactors, regardless of carbon source. Because of its very low cost compared to the other sources tested, molasses may be an attractive carbon source for denitrification if issues of ammonia production, turbidity and foaming can be resolved.     

Metabolic scope, swimming performance and the effects of hypoxia in the mulloway, Argyrosomus japonicus (Pisces: Sciaenidae)

The culture of the mulloway (Argyrosomus japonicus), like many other Sciaenidae fishes, is rapidly growing. However there is no information on their metabolic physiology. In this study, the effects of various hypoxia levels on the swimming performance and metabolic scope of juvenile mulloway (0.34 ± 0.01 kg, mean ± SE, n = 30) was investigated (water temperature = 22 °C). In normoxic conditions (dissolved oxygen = 6.85 mg l^− 1), mulloway oxygen consumption rate (M·o₂) increased exponentially with swimming speed to a maximum velocity (U_crit) of 1.7 ± < 0.1 body lengths s^− 1 (BL s^− 1) (n = 6). Mulloway standard metabolic rate (SMR) was typical for non-tuna fishes (73 ± 8 mg kg^− 1 h^− 1) and they had a moderate scope for aerobic metabolism (5 times the SMR). Mulloway minimum gross cost of transport (GCOT_min, 0.14 ± 0.01 mg kg^− 1 m^− 1) and optimum swimming velocity (U_opt, 1.3 ± 0.2 BL s^− 1) were comparable to many other body and caudal fin swimming fish species. Energy expenditure was minimum when swimming between 0.3 and 0.5 BL s^− 1. The critical dissolved oxygen level was 1.80 mg l^− 1 for mulloway swimming at 0.9 BL s^− 1. This reveals that mulloway are well adapted to hypoxia, which is probably adaptive from their natural early life history within estuaries. In all levels of hypoxia (75% saturation = 5.23, 50% = 3.64, and 25% = 1 .86 mg l^− 1), M·o₂ increased linearly with swimming speed and active metabolic rate (AMR) was reduced (218 ± 17, 202 ± 14 and 175 ± 10 mg kg^− 1 h^− 1 for 75%, 50% and 25% saturation respectively). However, U_crit was only reduced at 50% and 25% saturation (1.4 ± < 0.1 and 1.4 ± < 0.1 BL s^− 1 respectively). This demonstrates that although the metabolic capacity of mulloway is reduced in mild hypoxia (75% saturation) they are able to compensate to maintain swimming performance. GCOT_min (0.09 ± 0.01 mg kg^− 1 m^− 1) and U_opt (0.8 ± 0.1 BL s^− 1) were significantly reduced at 25% dissolved oxygen saturation. As mulloway metabolic scope was significantly reduced at all hypoxia levels, it suggests that even mild hypoxia may reduce growth productivity.     

Physiological changes in rainbow trout held under crowded conditions and fed diets with different levels of vitamins E and C and highly unsaturated fatty acids (HUFA)

Rainbow trout (Oncorhynchus mykiss) maintained in crowded (100 kg m^− 3) and uncrowded (20 kg m^− 3) conditions were fed 42 days with five experimental diets having different levels of vitamin E (25.6 and 275.6 mg kg diet^− 1), C (0 and 1000 mg kg diet^− 1) and HUFA (highly unsaturated fatty acids, 12.5 and 320.5 g kg diet^− 1): −E−HUFA, −E+HUFA, +E−HUFA, +E+HUFA, −C+E+HUFA. Cortisol, plasma metabolites, tissue glycogen, fish composition, and tissue fatty-acid profile were evaluated at the end of the experimental period. In general, no changes in cortisol levels were associated with crowding, although +E+HUFA and −C+E+HUFA fish showed higher levels (mean ± SE, 55.5 ± 11.1 and 78.0 ± 11.3 ng ml^− 1) as a consequence of a possible interaction between chronic crowding and diet composition. Protein and glucose con-centration in plasma displayed no effect of crowding, but liver glycogen showed a general tendency to decrease in −E−HUFA, −E+HUFA, +E−HUFA, +E+HUFA, −C+E+HUFA crowded groups (70.2 ± 2.1, 52.1 ± 2.5, 73.4 ± 7.4, 91.7 ± 3.3, 74.2 ± 8.4 mg g^− 1 tissue, respectively) compared to uncrowded groups (108.9 ± 14.2, 82.7 ± 8.8, 92.4 ± 10.7, 99.1 ± 10.0, 103.5 ± 15.6 mg g^− 1 tissue, respectively), thus proving significant in −E+HUFA fish. Variations in total lipids, triglycerides, total cholesterol and HDL as well as LDL cholesterol in plasma were manifested under crowding conditions, displaying a certain influence of vitamin E and HUFA dietary content. Final body composition, in general, showed no change attributable to fish density, but some differences associated with diet composition were found in lipid and moisture percentages of crowded fish. Liver and muscle fatty-acid profile revealed a clear effect of the dietary lipid source that was more evident in muscle than in liver at normal fish density, and in some cases this effect was modulated by dietary vitamin E and C content and fish-culture conditions.     

A laboratory scale recycling water unit for tilapia breeding

The technical features of a laboratory scale water recycling unit for experimental small scale tilapia breeding are described. Two units (1 and 2) were operated during a 6 month period, carrying a similar fish load (7·5 kg) and feeding rate (2% fish body weight/day). Unit 1 received natural illumination, while unit 2 was artificially illuminated (14/10 - light/dark cycle). Both units were equipped with a biological filter bed (substrate surface area, 3500 cm²). In unit 1, total ammonium and nitrite concentrations ranged from 0·05 to 0·5 mg liter⁻¹, while nitrate varied between 10–40 mg liter⁻¹. In unit 2 corresponding values were 0·15-3 mg liter⁻¹, 0·05–0·8 mg liter⁻¹ and 10–40 mg liter⁻¹. Temperatures ranged between 20–29°C and pH values between 7·5–6·9 in both units. Dissolved oxygen concentrations decreased gradually from 5·6 to 3·4 mg liter⁻¹ in unit 1 and from 5·6 to 2·6 mg liter⁻¹ in unit 2. Twenty-six spawnings occurred in unit 1 in March and April, while only eight spawnings occurred in unit 2, possibly because of the absence of sunlight. The significance of these results are discussed.     

Growth and survival of larval and juvenile cobia Rachycentron canadum in a recirculating raceway system

Cobia Rachycentron canadum is a fast-growing, pelagic marine species that has recently attracted aquaculturists in both the research and commercial sectors. The typical method of grow-out for this species is in outdoor systems where production is limited to locations and seasons conducive for adequate growth and survival. Expanding the culture of cobia to indoor recirculating aquaculture systems (RAS) would allow for the production of fingerlings throughout the year and extend production to cooler regions. Two rearing trials were conducted to examine the growth and survival of cobia from hatching through 4 (trial 1, T1) or 35 (trial 2, T2) g in RAS. Cobia larvae were reared in circular tanks placed in a raceway to control water temperature and quality. During early juvenile grow-out, fish were transferred without grading to a second raceway on 29 dph (T1) or over a period of grading from 29–43 dph (T2). Larval growth (1–22 dph) measured as standard length was similar for both trials ranging from  3.9 to 14.7 mm. However, larval growth measured as wet weight (0.033 g, T1; 0.026 g, T2) or dry weight (5.7 mg, T1; 3.9 mg, T2) was significantly greater on 22 dph during T1 as was the ratio between myotome height and standard length. These differences may have resulted from an increase in initial densities from 8.7 larvae l^− 1 (T1) to 14.7 larvae l^− 1 (T2) which apparently caused an increase in food competition and overall aggression. During juvenile grow-out, cobia reached 4.0 g on 43 dph in T1 and 35.4 g on 71 dph in T2 matching weights achieved during grow-out in outdoor ponds. Over the course of both trials, survival was similar to that reported in outdoor ponds. Mean survival (± S.D.) during the early rearing phase (hatching through 29 or 43 dph) averaged 13.2 ± 3.2 % and 10.4 ± 3.2 % corresponding to final densities of 0.9 ± 0.2 and 1.2 ± 0.4 fish/l for T1 and T2, respectively. During the first grow-out phase (29–43 dph), survival of fish moved into the open raceway was 64.5% in T1 and 88.7 % in T2. Survival of cobia during the second grow-out phase (43–71 dph) for T2 was 92.5%. The results of this study indicate that cobia can be successfully cultured in indoor systems from hatching through at least 35 g without negatively affecting growth or survival.     

Effect of increasing salinity on physiological response in juvenile scallops Argopecten purpuratus at two rearing temperatures

Argopecten purpuratus can be cultivated using Recirculating Aquaculture Systems (RAS) as a method to increase production. In order to determine physiological response of A. purpuratus under different salinities and temperature conditions, two groups of juvenile scallops (small: h = 6.5 mm and large: h = 25.5 mm) were acclimated and close-cultured at salinities of 34, 38, and 42 g/l, at 16 and 22 °C and fed on Isochrysis galbana and Chaetoceros calcitrans. Survival, shell growth and scope for growth were determined at the end of the trials. Survival showed an inverse relationship with temperature and ammonia levels. In small scallops an increase in salinity at 16 °C increased survival. However, this relationship was not evident at 22 °C. On the other hand, salinity did not affect survival of large juveniles. Small juveniles had a lower survival (approximately 40%) than larger scallops (up to 85%) throughout the trials. Oxygen consumption was not affected by salinity. Small scallops showed similar oxygen consumption at 16 and 22 °C but in large juveniles higher values were registered at 22 °C. In large juveniles routine consumption at 16 °C was higher (up to 35%) than standard consumption. This pattern was not evident at 22 °C, suggesting that oxygen demand is higher regardless of feeding condition. NH₄⁺–N excretion rate is inversely related to salinity. Only small juveniles showed a higher NH₄⁺–N excretion rate at 22 °C. Scope for growth was positive in all treatments, although the upper limit of salinity should not be based only in this index. Higher scope for growth values at 38 and 42 g/l was related with a reduction in ammonia excretion and high absorption efficiency. In addition, an increase in salinity produced a reduction in NH₃–N proportion and under hypersaline conditions scallops tended to decrease excretion as a way of osmoconformation. This explains our findings of higher survival rates at higher salinities. Even though the scope for growth is positive at 42 g/l, the osmotic stress reduces the survival chances. The data obtained can be considered useful information for A. purpuratus culture under controlled conditions.     

Efficacy of bithionol as an oral treatment for amoebic gill disease in Atlantic salmon Salmo salar (L.)

This study examined the efficacy of bithionol as an oral treatment for Atlantic salmon Salmo salar affected by amoebic gill disease (AGD). The current commercial management strategy of AGD is a costly 3 h freshwater bath. It is labour intensive and the number of baths needed appears to be increasing; hence, there is an effort to identify alternative treatments. Efficacy was examined by feeding AGD-affected Atlantic salmon twice daily to satiation with bithionol, an antiprotozoal, at 25 mg kg^− 1 feed. Three seawater (35‰, 17 °C) re-circulation systems were used each consisting of three tanks containing 32 Atlantic salmon smolts with an average (± SEM) mass of 90.4 g (± 5.2). Three feeds were examined in the trial including bithionol, plain commercial control and oil coated commercial control. Feeding commenced 2 weeks prior to exposure to Neoparamoeba spp. at 300 cells L^− 1 and continued for 28 days post-exposure. Efficacy was determined by examining gross gill score and identifying percent lesioned gill filaments twice weekly for 4 weeks post-exposure. Bithionol when fed as a two-week prophylactic treatment at 25 mg kg^− 1 feed delayed the onset of AGD pathology and reduced the percent lesioned gill filaments by 53% and halved the gill score from 2 to 1 when compared with both the plain and oil controls during an experimental challenge. There were no palatability problems observed with mean feed intake of bithionol over the trial duration with fish consuming higher levels of the bithionol diet compared to both the oil and plain controls. This study demonstrated that bithionol at 25 mg kg^− 1 feed, when fed as a two-week prophylactic treatment for Neoparamoeba spp. exposure, delayed and reduced the intensity of AGD pathology and warrants further investigation as an alternative to the current freshwater bath treatment for AGD-affected Atlantic salmon.     

Filtration of king scallops (Pecten maximus)

Filtration rates of hatchery-reared king scallop (Pecten maximus L.) juveniles, fed a single species alga diet (Pavlova lutheri (Droop) Green), were measured at a range of temperatures (6–21 °C). Weight specific filtration rate (ml min⁻¹ g⁻¹ (live weight)) of juveniles of a selected size range of 17–19 mm shell height (0.26–0.36 g live weight) increased with temperature above 16 °C and decreased below 11 °C, but was not significantly different between these two temperatures. Measurements at 16 °C using juveniles with a wider size range of 10–25 mm shell height (0.05–0.8 g live weight) gave the allometric equation: filtration rate (ml min⁻¹)=12.19×weight (g)^0.887. Filtration rate decreased significantly when the cell concentration was greater than 200 cells μl⁻¹ (4.25 mg (organic weight) l⁻¹). With six other algae food species, filtration rates similar to those with P. lutheri were only achieved with Chaetoceros calcitrans (Paulsen) Takano. All other algae species tested were cleared from suspension at significantly lower rates. Experiments with diet mixtures of P. lutheri and these other algae suggested that this was usually a reflection of lowered filtration activity, rather than pre-ingestive rejection of cells. In experimental outdoor nursery rearing systems, the filtration rate was inversely proportional to the concentration of cells in the inflow, in the range 5–210 cells μl⁻¹. It was not affected by flow rate (2–130 l h⁻¹, equivalent to 0.12–28.38 l h⁻¹ g⁻¹ (live weight)) with scallop juveniles stocked from 2 to 62 g l⁻¹. The results are discussed in relation to on-growing scallops at field sites.     

Comparison of nutrients release among some maricultured animals

Integrated mariculture is a feasible method to maintain sustainable and high productivity of aquaculture. The choice of cultured animals and biofilters in the integrated system has to be made on the basis of their nutrient release rates and the clearance rate of each component of the system. We are examining the nutrient release rates among fish (mangrove snapper, Lutjanus russeli, and sea perch, Abudefduf septemfasciatus), abalone (Haliotis diversicolor), scallops (Chlamys noblis), and green mussels (Perna viridis) in the laboratory. Fish feed is the major sources of inorganic nutrient input in fish farms. The orthophosphate and ammonia release rates of minced trash fish (1593 μg P g⁻¹ day⁻¹ and 150 μg N g⁻¹ day⁻¹) were respectively 6–12 times and 4–88 times higher than those of cultivated fish. Mangrove snapper had the overall highest nutrient release rate, followed by sea perch, abalone, scallops, and mussels for nitrite and nitrate; and followed by abalone, sea perch, mussels, and scallops for orthophosphate and ammonium. Among mollusks, abalone had the highest orthophosphate (162 μg P g⁻¹ day⁻¹), nitrate (1.4 μg N g⁻¹ day⁻¹), nitrite (1.6 μg N g⁻¹ day⁻¹) and ammonium (25.0 μg N g⁻¹ day⁻¹) release rates per gram wet weight per day. Abalone released large amounts of orthophosphate, nitrite and nitrate in the experiment. Scallops and green mussels had low nutrient release rates.     

Outdoor phytoplankton continuous culture in a marine fish–phytoplankton–bivalve integrated system: combined effects of dilution rate and ambient conditions on growth rate, biomass and nutrient cycling

Natural phytoplankton populations were cultured in outdoor continuous cultures using fish-farm effluents as the source of nutrients. The dilution rate was assumed to be the integrating factor of phytoplankton growth and biomass development (flux and stock). In this context, the combined effects of (i) dilution rates of the outdoor culture and (ii) ambient conditions were tested on phytoplankton growth, biomass and cycling of the major nutrient elements (C, N and P). Experiments were carried out in outdoor polyester tanks (0.7 m deep), homogenised by gentle aeration. Si/P ratio was balanced at around 5 in the inflow in order to induce diatom domination while maintaining high N and P assimilation by phytoplankton. Nutrient cycling was assessed through analyses of the different forms of particulate and dissolved nutrients in the inflow and the outflow. Culture dilution rates determined the longevity of the culture and the assimilation efficiency of nutrients. Dissolved phosphorus was the most limiting nutrient. The optimal dilution rate was approximately 0.5 day⁻¹ at 10 °C and 1.5 day⁻¹ at 20 °C with a mean diatom biomass of 9 μM P. Under these conditions, 80% of the dissolved nutrients provided to the tanks were transformed, a production of 8 g C m⁻² day⁻¹ and an assimilation rate of 0.3 g P m⁻² day⁻¹ were recorded. Assimilation by diatoms was the major pathway of nutrient cycling. During the experiment, a bottom sediment developed progressively and this also played an important role in denitrifying the excess dissolved nitrogen in the fish-farm effluent. However, the results showed that diatom biomass can collapse and we hypothesize that this was the consequence of an increase in cellular sinking rates due to cell aggregation under nutrient or light stress. Modelling approaches are needed in future research in order to determine optimal dilution rates taking into account phytoplankton growth rates, nutrient inputs and ambient conditions (e.g. light and temperature).     

Substitution of Chaetoceros muelleri by Spirulina platensis meal in diets for Litopenaeus schmitti larvae

The nutritional response of Litopenaeus schmitti larvae to substitution of Chaetoceros muelleri by Spirulina platensis meal (SPM) was evaluated. The substitution levels (S) were 0%, 25%, 50%, 75% and 100%, dry weight basis. Final larval length (FL) ranged from 1.98 to 3.16 mm for the different substitution levels. There was a significant relationship between S and FL, described by the following quadratic equation: FL = 2.853 + 0.01598S − 0.000233S². The substitution level (S) yielding maximum FL was 34.2%. Development index (DI) values ranged from 2.84 to 3.93 and were dependent on substitution level. The corresponding equation was DI = 3.799 + 0.00945S − 0.000189S² (P < 0.01). Maximum DI was obtained at 25.0% substitution. Survival was high (82–87%) and no significant differences were found between treatments. Protein digestibility of either microalgae was high, with 92% for SPM and 94% for C. muelleri, with no significant differences between them. The results in this study indicate that an adequate balance of nutrients in relation to the requirements of the species is critical. To simultaneously improve FL and DI, a 30% substitution of C. muelleri by SPM is suggested. This is equivalent to feeding 0.15 mg larvae^− 1 day^− 1 dry weight basis of a 70% C. muelleri/30% SPM diet, representing 0.078 mg protein larvae^− 1 day^− 1, 0.026 mg lipids larvae^− 1 day^− 1 and 2.732 J larvae^− 1 day^− 1.     

Effect of low water temperature on viral replication of white spot syndrome virus in Procambarus clarkii

This study evaluated the effect of low water temperature (10 ± 1 °C) on viral infection and replication of white spot syndrome virus (WSSV) in crayfish, Procambarus clarkii, under standardized conditions. Crayfish were (i) maintained at 24 ± 1 °C before challenge and 10 ± 1 °C afterwards, or (ii) maintained at 10 ± 1 °C before challenge and 24 ± 1 °C afterwards. No mortality was observed when crayfish were held at 10 ± 1 °C after challenge, but mortality reached 100% when they were transferred to 24 ± 1 °C. Competitive PCR showed that viral levels at 10 ± 1 °C rose from 10⁶ to 10⁸ copies/mg of gill tissues, while at 24 ± 1 °C levels increased from 10⁶ to 10¹⁰ copies/mg of gill tissues during the same time interval. These results showed that a low water temperature of 10 ± 1 °C could reduce viral replication when compared to 24 ± 1 °C but could not prevent it.     

Breeding and juvenile culture of the lined seahorse, Hippocampus erectus Perry, 1810

All seahorse species worldwide have been placed under CITES Appendix II since 2004, because they have been over-exploited for traditional Chinese medicine and aquarium trades. Aquaculture has been recognized as a long-term solution for sustaining the seahorse trade while minimizing wild collection. In this study, we evaluated the breeding and juvenile culture of an important aquarium seahorse species, the lined seahorse Hippocampus erectus, Perry 1810. Pairing, mating and copulation behavior were observed. Gestation time and brood size were 17.33 ± 2.94 days and 272.33 ± 66.45 individuals/brood, respectively. Growth rates differed among juveniles from different broods. Effects of temperature on the growth rates and survivorship of the juveniles during the first two weeks were compared. The highest growth rate and survivorship of the juveniles occurred at 28–29 °C among the temperatures tested (24–33 °C). Growth rate and survivorship of the juveniles during the first 9 weeks at 28 °C were investigated. The final standard length and survivorship of the juveniles were 6.32 ± 0.52 cm and 71.11 ± 10.18%, respectively, and the relationship between the wet weight and the standard length of the juvenile seahorses can be expressed as: W = 0.0034 L^2.5535 (r² = 0.9903, n = 12, P < 0.01). These findings suggest that H. erectus is a good candidate for commercial aquaculture.     

Serotonin induces ovarian maturation in giant freshwater prawn broodstock, Macrobrachium rosenbergii de Man

This study investigated the effects of serotonin (5-hydroxytryptamine or 5HT) on ovarian development in Macrobrachium rosenbergii de Man. Adult female prawns at the ovarian stage I (spent) were injected with 5HT at 1, 5, 10, 20 and 50 μg g^− 1 body weight (BW) intramuscularly on days 0, 5 and 10, and sacrificed on day 15. The doses as related to the effect could be categorized into three levels: low (1 and 5 μg g^− 1 BW of 5HT), medium (10 and 20 μg g^− 1 BW of 5HT) and high (50 μg g^− 1 BW of 5HT). The low-dose, especially at 1 μg g^− 1 BW, caused prawns to exhibit a significant increase in ovarian index (ovarian weight/body weight × 100) (5.79 ± 0.09%) as compared to the control (1.49%). The ovaries of most of these prawns could develop to stage IV (mature) and contained synchronously mature oocytes while most of the control ovaries remained at stage I and II (proliferative), and contained only oogonia to previtellogenic (Oc1, Oc2) and early vitellogenic oocytes (Oc3). The medium- and high-dose treated prawns exhibited ovaries that could reach stages III and IV and contained various types of oocytes of different maturity. Pretreatment with 5HT receptor antagonist, cyproheptadine (CYP), at 10 μg g^− 1 BW before 5HT injection significantly suppressed the effect of 5HT. Intramuscular injection of the 5HT-primed thoracic ganglion culture medium into CYP-pretreated prawns resulted in the increase of ovarian index about 5–6 times more than in the control, and in the groups injected with 5HT-primed media from muscle strip, eyestalk and brain. The ovaries of most prawn could develop up to stage IV and contained synchronously developed vitellogenic (Oc4) and mature oocytes (Oc5). These findings suggest that 5HT indirectly induces ovarian development and oocytes maturation in M. rosenbergii, probably via a putative ovarian stimulating factor released from the thoracic ganglia.     

Effects of body weight, water temperature and ration size on ammonia excretion by the areolated grouper (Epinephelus areolatus) and mangrove snapper (Lutjanus argentimaculatus)

The effects of body weight, water temperature and ration size on ammonia excretion rates of the areolated grouper Epinephelus areolatus and the mangrove snapper Lutjanus argentimaculatus were investigated. Under given experimental conditions, L. argentimaculatus had a higher weight-specific ammonia excretion rate than E. areolatus. Weight-specific ammonia excretion rates of fasted individuals of both species showed an inverse relationship with body weight (W, g wet wt.), but a positive relationship with water temperature (t, °C). The relationships for total ammonia nitrogen (TAN) were: E. areolatus: TAN (mg N kg⁻¹ d⁻¹)=21.4·exp^0.11t·W^−0.43 (r²=0.919, n=60); L. argentimaculatus: TAN (mg N kg⁻¹ d⁻¹)=121.5·exp^0.12t·W^−0.55 (r²=0.931, n=60). Following feeding, the weight-specific ammonia excretion rate of E. areolatus increased, peaked at 2 to 12 h (depending on temperature), and returned to pre-feeding levels within 24 h. A similar pattern was observed for L. argentimaculatus, with a peak of TAN excretion being found 6 to 12 h after feeding. Stepwise multiple regression analysis indicated that weight-specific TAN excretion rates of both species increased with increasing temperature and ration (R, percent body wt. d⁻¹): E. areolatus: TAN (mg N kg⁻¹ d⁻¹)=22.8·t−28.8·R−378.2 (r²=0.832, n=24); L. argentimaculatus: TAN (mg N kg⁻¹ d⁻¹)=22.9·t−25.4·R−216.4 (r²=0.611, n=24). The effect of body weight on weight-specific postprandial TAN excretion was not significant in either species (p>0.05). This study provides empirical data for estimating ammonia excretion of these two species under varying conditions. This has application for culture management.     

Energy and nitrogen partitioning in 250 g Atlantic cod (Gadus morhua L.) given graded levels of feed with different protein and lipid content

The effects of feed intake level on energy and nitrogen partitioning were studied in juvenile Atlantic cod (250 g) fed two fish meal based diets differing in protein and lipid content (54:31 and 65:16) at 10 °C. Replicate groups of cod were feed deprived for 32 days or fed one of the two diets at 25, 50, 75 or 100% of group satiation for 60 days. Feed intake and oxygen consumption were measured daily and weights and chemical composition of carcass, liver, viscera and whole body were measured at start and end. Diet digestibilities were assessed in a separate experiment.

The whole body and carcass growth rates at a given feed intake did not differ between dietary groups, but the liver grew faster in the fish fed the low protein diet, resulting in higher hepatosomatic indices at the end of the experiment in the groups fed this diet.

The efficiency of utilisation of digestible nitrogen for growth (k_DNg) was higher for the low protein diet (0.73 ± 0.02) than for the high protein diet (0.53 ± 0.05), resulting in higher nitrogen retention at a given nitrogen intake. No difference in percentage nitrogen retention was seen in full-fed fish however (31.2 ± 2.5 and 28.4 ± 1.6% for the low protein and high protein diets, respectively). This can be explained by higher nitrogen intake in the fish fed the high protein diet, resulting in a smaller proportion of the intake being used for maintenance.

There was no difference in energy utilisation between dietary groups. The digestible energy requirement for maintenance (DE_maint) was 53.8 ± 0.9 kJ kg^− 1 d^− 1 (42.3 ± 0.7 kJ kg^− 0.8 d^− 1) and the utilisation efficiency for growth (k_DEg) was 0.80 ± 0.02. The energy retention in full-fed fish was 31.3 ± 3.5 and 31.7 ± 1.0% for the low protein and high protein diets, respectively. The deposited energy was distributed in approximately equal proportions in the liver and carcass, whereas viscera accounted for a minor proportion. At a given energy intake, the fish fed the high protein diet deposited more energy in the carcass and less in the liver than did those fed the low protein diet.