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.     

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.     

Appetite, gut transit, oxygen uptake and nitrogen excretion in captive Atlantic halibut, Hippoglossus hippoglossus L., and lemon sole, Microstomus kitt (Walbaum)

Atlantic halibut, Hippoglossus hippoglossus L., eat larger satiation meals (mean 11.7% body weight) than lemon sole, Microstomus kitt (Walbaum), (2.6% body weight). Total gut clearance time was about 120 h for halibut and 72 h for lemon sole. There are marked differences in feeding behaviour between the two species; halibut feed in midwater and require several body lengths of approach swimming before taking large items of food, while lemon sole eat only off the bottom. In shared tanks, no aggressive interaction was observed. A duoculture system holding small numbers of lemon sole with the more valuable halibut is recommended as a means of minimizing food waste and tank fouling. Oxygen uptakes of 0.07-0.11 ml O₂ g fish wt⁻¹ h⁻¹ (depending on nutritional state) were recorded for the two species. Ammonia nitrogen outputs were also similar. Starved halibut excreted 2.32 μg N g⁻¹ h⁻¹, fed animals 5.08 μg N g⁻¹ h⁻¹. The corresponding values for lemon sole were 3.26 μg N g⁻¹ h⁻¹ and 6.37 μg N g⁻¹ h⁻¹, respectively.     

An improved enzyme preparation for rapid mass production of protoplasts as seed stock for aquaculture of macrophytic marine green algae

Preparation of protoplasts and their subsequent applications for both basic and applied research of marine macroalgae remains largely under developed due to lack of development of reliable methods with consistent yields of viable protoplasts. An improved enzyme preparation with a single commercial enzyme, e.g. 2% Cellulase Onozuka R-10 in 1% NaCl solution, was developed to produce protoplasts rapidly from different green algal genera of Ulva, Enteromorpha and Monostroma. The simple dissolution of enzyme powder in 1% NaCl resulted in exclusion of 2% Macerozyme R-10 from the mixture consisting of 2% Cellulase Onozuka R-10 with 3% NaCl earlier reported as superior for the same algae. Optimal conditions for the isolation of maximum yields of viable protoplasts were found to be with 2% Cellulase Onozuka R-10 incubated at 20 °C for 2 h in 1% NaCl solution with 0.8 M mannitol adjusted to pH 6.0. The protoplast yield with optimized enzyme mixture was as high as 102.8 × 10⁶ cells g^− 1 f. wt for M. oxyspermum while it was in the range of 74.4–88.6 × 10⁶ cells g^− 1 f. wt thallus for seven species of Ulva, and 82.5–95.4 × 10⁶ cells g^− 1 f. wt for three species of Enteromorpha. The regeneration rate of protoplasts isolated using this method ranged from 89 to 92% with normal morphogenesis. The seeding of nylon threads with isolated protoplasts of M. oxyspermum was successful and after 3–4 weeks the entire frame with nylon threads became thick green in color with tiny germlings in laboratory culture. Thus, the method described in the present study allow for rapid mass production of viable protoplasts that could be potentially used as a source for seed material for mariculture and for other applied phycological research.     

Impact of starvation and handling stress on rigor development and quality of Atlantic salmon (Salmon salar L)

Farmed Atlantic salmon (Salmo salar L) were fed to satiation or starved for 35 d, and subjected to acute pre-slaughter stress or careful handling before slaughtering. Rigor development, post-mortem energy metabolism and quality variations were analysed during 72 h cold storage. The body weight averaged 2.9 and 3.5 kg of the fed and starved salmon, respectively. The starved salmon had lower condition factor (0.99 vs. 1.11) and higher slaughter yield (92.6 vs. 87.5%), but the fat content was similar (11.8–12.9%) in the fed and starved salmon. The initial glycogen level was lower in the starved salmon (21 vs. 29 μmol g^− 1), whereas initial ATP levels (6.4–6.6 μmol g^− 1) were not affected by starvation. Pre-slaughter stress accelerated rigor development, stimulated lactate formation through post-mortem glycolysis, and increased the breakdown of ATP and CP. The effect of stress on rigor development was less pronounced in starved salmon, and a significant interaction was observed between nutritional status and stress. Rigor development was closely correlated to ATP breakdown, especially in fillets. Acute stress accelerated flesh softening during ice storage. After 72 h storage, the fillets of the starved salmon exposed to careful slaughter handling had the significantly firmest texture, whereas the most intense fillet colour was observed in the starved/stressed salmon. Liquid leakage from the muscle during cold storage was lower in the starved salmon, but pre-slaughter stress had no significant impact. It is concluded that a starvation period of five weeks can improve the resistant to acute stress prior to slaughtering of Atlantic salmon that in turn hampers rigor development.     

Shipping studies with juvenile and adult Malaysian prawns Macrobrachium rosenbergii (de Man)

Four studies examined shipping factors of packing technique, density, duration, type of water and use of habitat material for shipping juvenile and adult prawns Macrobrachium rosenbergii. Prawns were shipped in double polyethylene lined 38 × 38 × 20 cm deep styrofoam boxes containing oxygenated water. At temperatures of 19–20°C, 17 g prawns could be shipped safely for 42 h at a density of 10–12 prawns per box (12–15 g liter⁻¹ shipping water). Juveniles, mean size about 6 g, could be shipped at a density of 40 per box (18 g liter⁻¹) for 24 h or 20–25 per box (9–11 g liter⁻¹) for 48 h. Use of mesh material to increase surface area in the box did not appear beneficial nor did shipping in brackish water (salinity8‰). Adults packed unrestricted resulted in survival rates substantially higher than those obtained from immobilized prawns wrapped in mesh.

During the shipment, pH and dissolved oxygen concentrations decreased whereas ammonia concentrations increased. The decreased pH levels may have reduced the ammonia toxicity by decreasing the amount of toxic unionized ammonia (NH₃) in solution. In general, dissolved oxygen concentrations appeared more closely related to survival rates than did other water quality parameters which were measured.     

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.     

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.     

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.     

Furunculosis in Atlantic salmon (Salmo salar L.) is not readily controllable by bacteriophage therapy

The potential of bacteriophage therapy to control bacterial disease in farmed fish was tested using, as an example, furunculosis of Atlantic salmon, caused by Aeromonas salmonicida subsp. salmonicida.

In vivo testing with Atlantic salmon and rainbow trout (Oncorhynchus mykiss Walbaum) showed no adverse effects, with bacteriophage generally cleared within 96 h of administration by either intraperitoneal (i.p.) injection or oral in-feed.

Juvenile Atlantic salmon were administered a combination of bacteriophage O, R and B (1.9 × 10⁸ pfu fish^− 1) by i.p. injection, after they had been challenged with A. salmonicida subsp. salmonicida 78027, also by i.p. injection. The fish that were injected with bacteriophage immediately after challenge died at a significantly slower rate then those that were either not treated with bacteriophage, or treated 24 h post-challenge. However, the end result (100% mortality) was not affected.

In further experiments the effects of oral (1.88 × 10⁵ pfu g^− 1 fish^− 1 daily for 30 days), bath (1.04 × 10⁵ ml^− 1 daily for 30 days) and i.p. (6.25 × 10⁷ pfu fish^− 1) phage treatment to control furunculosis in experimentally infected Atlantic salmon were compared with antibiotherapy (treatment with 10 mg kg^− 1 bw^− 1 day^− 1 oxolinic acid for 10 days), using an indirect cohabitation challenge. No protection was offered by any of the bacteriophage treatments, compared to the positive challenge group, although significant protection was offered by the oxolinic acid treatment. Analysis of samples taken from the trials demonstrated that bacteriophage were correctly administered to the fish and, on occasion, were isolated from fish that had succumbed to furunculosis. It was also shown that bacteriophage resistant A. salmonicida subsp. salmonicida isolates could be recovered from mortalities in all the treatment groups.

The results suggest that, although there were no safety problems associated with the approach, furunculosis in Atlantic salmon is not readily controllable by application of bacteriophage.     

Classification of differentiating oocytes during ovarian cycle in the giant freshwater prawn, Macrobrachium rosenbergii de man

Based on the light microscopic observations of cells' sizes, chromatin patterns, amount of lipid droplets and yolk granules, the female germ cells could be classified into four different phases, which include 1) oogonia (Oog), 2) primary oocytes (pOc), 3) secondary oocytes (sOc), and 4) mature oocyte (mOc). Oog are small oval-shaped cells with irregular-shaped nuclei sizing 4–6 μm in diameter. They rest on the connective tissue germinal cord at the tip of each ovarian pouch (lobule). Oogonia increase their number through mitotic division, and the daughter cells move into ovarian pouch where they undergo first meiotic division to become primary oocytes, which have various steps of 1st meiotic prophase accumulating at the innermost zone of the ovarian pouch. The primary oocytes are small oval-shaped cells (8.5–10 μm in diameter) with large nuclei containing chromatin in various states of condensation that finally transform into chromatids. Their nuclei are surrounded by thin rim of faint blue-stained cytoplasm. The secondary oocytes derived from 2nd meiosis and comprise five steps: Oc1 and Oc2, classified as previtellogenic oocytes, Oc3 and Oc4, classified as vitellogenic oocytes, and mature oocyte (mOc) The zones of ovarian pouch are defined based on the accumulation of various steps of developing oocytes, namely, oogenic, previtellogenic, vitellogenic and mature zones, respectively. The ovarian cycle is divided into five stages based on the number and types of oocytes present in each stage. Stage 0 and I are spawn and spent stages. Stage II and III are proliferative and premature stages, while stage IV is mature stage. During ovarian stage I, each ovarian pouch contains primarily oogonia, primary oocytes, Oc1 and a few Oc2. In stage II, the pouch contains mainly Oc2 and Oc3, while in stage III the predominant cells are Oc4. Mature oocytes appear synchronously, in stage IV. The ovulating mature oocytes pass through the thin disrupted wall of ovarian pouch into subcapsular space, that leads into the oviduct situated on the ventro-lateral side of the ovarian lobe. At spawning, the ovarian pouches break down and only connective sheaths and hemolymph sinuses remain. The germinal cords and islets of oogonia remain in the central area of stage 0 ovary. The ovarian capsule, including the muscular layer, becomes attenuated as the ovary progresses from stage 0 to IV. The hemolymph vessels become highly convoluted in the central area of the ovary, and they branch radially into smaller hemolymph sinuses around each oogenic pouch.     

Effect of docosahexaenoic acid enrichment in Artemia on growth of Pacific bluefin tuna Thunnus orientalis larvae

Feeding enriched Artemia induces growth failure in Pacific bluefin tuna (PBT) Thunnus orientalis larvae; however, feeding of yolk-sac larvae of marine fish promotes larval growth. It is considered that this growth failure partly results from dietary docosahexaenoic acid (22:6n-3, DHA) deficiency. Therefore, we examined the effect of DHA contents in enriched Artemia on the growth of PBT larvae. Artemia nauplii were enriched with graded levels of DHA ethyl ester, and fed to PBT larvae for 9 days. Yolk-sac larvae of Japanese parrotfish Oplegnathus fasciatus were used as a reference diet. The DHA contents in Artemia increased with the enrichment from 0 mg g^− 1 dry weight basis (DW) to 25 mg g^− 1 DW, while the content in the reference diet was 21 mg g^− 1 DW. Feeding of enriched Artemia significantly improved the growth of PBT larvae. However, this improvement was negligible when compared with the excellent growth of the larvae that were fed the reference diet. PBT larvae that were fed the reference diet accumulated approximately twice or much higher levels of DHA in the neutral and polar lipids in the body when compared with the larvae that were fed enriched Artemia. These results show that PBT larval growth cannot be promoted by feeding enriched Artemia even if the DHA contents in Artemia are elevated to the same levels as those of yolk-sac larvae. The incorporation of dietary DHA into phospholipids in the fish body may be desirable for the normal growth of PBT larvae.     

Effects of dissolved oxygen on hemolymph parameters of freshwater giant prawn, Macrobrachium rosenbergii (de Man)

Profiles of changes in physiological parameters of freshwater giant prawns, Macrobrachium rosenbergii, exposed to various dissolved oxygen (DO) levels of 7.75, 4.75, 2.75, and 1.75 mg l⁻¹ are reported. The parameters involved in osmoregulation and oxygen transport were monitored for a 6-day period. Notable depressions in hemolymph osmolality, Na⁺, K⁺, and Cl⁻ contents were observed within 24 h after exposure to hypoxia at 2.75 and 1.75 mg O₂ l⁻¹, and thereafter remained at rather steady levels, which were significantly lower than those under normoxic conditions (4.75–7.75 mg O₂ l⁻¹). The extent of depression of osmotic-related constituents, hemolymph osmolality and Cl⁻ in particular, increased with decreased DO.

Oxyhemocyanin constituted 65.46–65.84% of total hemolymph proteins under the various DO levels examined; both hemolymph oxyhemocyanin and proteins showed notable elevations 24 h after exposure to hypoxic conditions, and reached the highest and constant level by 48 h after exposure. The compensatory responses of prawns to reduced O₂ were manifested by increased O₂ uptake through augmentation of hemocyanin, which results in enhancement of oxygen binding capacity of the hemolymph. In the same period, a significant surge of the respiratory products, PCO₂ and HCO₃⁻, was also demonstrated 6 h after exposure to hypoxic conditions which resulted in hemolymph alkalosis. These processes likely resulted in an increase in water influx and consequent declines in hemolymph osmolality and ion composition. Furthermore, hyperventilation and respiratory alkalosis, indicated by increased oxyhemocyanin and pH, respectively, were found to be predominant responses of M. rosenbergii to hypoxic stress.     

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.     

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.     

Comparative performance of Gamma Amino Butyric Acid (GABA) and 5-Hydroxytryptamine (5-HT) in the diet of larvae and post larvae of giant freshwater prawn, Macrobrachium rosenbergii: Effect of dose and route of administration on growth and survival

Three experiments were conducted to evaluate the effect of different dose and route of administration of 5-Hydroxytryptamine (5-HT) and Gamma Amino Butyric Acid (GABA) on growth, survival and pigmentation of giant freshwater prawn, Macrobrachium rosenbergii, larvae (experiment 1) and post larvae (experiment 2 and 3). In experiment 1 larva were stocked at 100 /L in seven different treatments with each of three replicates. The treatments were T1 (control), T2 (100 μg/ml of 5-HT bath exposure for 2 days), T3 (1 μM of GABA bath exposure for 2 days), T4 (0.5% 5-HT in feed), T5 (0.25% 5-HT in feed), T6 (0.5% GABA in feed) and T7 (0.25% GABA in feed). Highest growth, transformation rate, pigmentation and survival of larvae were recorded in T2 group in experiment 1. In experiment 2, post larvae were stocked at 70 per tank with 200 L of water for 45 days. This experiment consisted of 5 treatment groups with each of 3 replication viz. T1 (control), T2 (0.5% 5-HT in feed), T3 (0.25% 5-HT in feed), T4 (0.5% GABA in feed) and T5 (0.25% GABA in feed). At the end of this experiment growth was found to be higher in the control than the other treatment group showing inhibitory effect of 5-HT and GABA on growth of post larvae. To confirm the result, a third experiment was conducted for 30 days. Thirty numbers of post larvae having similar size were segregated from the previous experimental tanks (experiment 2) and stocked in the tanks of 100 L of water for experiment 3. All treatments were fed with control diet. There was similar growth pattern in all the treatments, which were higher (P < 0.05) than control, confirms the inhibitory effect of neurotransmitter in the diet of PL. Therefore, the overall results of the present study suggest that the bath treatment of neurotransmitter is superior to the dietary addition with regard to the growth, survival and pigmentation of M. rosenbergii larvae. 5-HT is more effective than the GABA for larvae of M. rosenbergii.     

Replacement of fish meal by meat and bone meal in diets for large yellow croaker, Pseudosciaena crocea

A growth experiment was conducted to investigate the effects of replacement of fish meal (FM) by meat and bone meal (MBM) in diets on the growth and body composition of large yellow croaker (Pseudosciaena crocea). Six isonitrogenous (43% crude protein) and isoenergetic (20 kJ g^− 1) diets replacing 0, 15, 30, 45, 60 and 75% FM protein by MBM protein were formulated. Each diet was randomly allocated to triplicate groups of fish in sea floating cages (1.0 × 1.0 × 1.5 m), and each cage was stocked with 180 fish (initial average weight of 1.88 ± 0.02 g). Fish were fed twice daily (05:00 and 17:30) to apparent satiation for 8 weeks. The water temperature ranged from 26.5 to 32.5 °C, salinity from 32 to 36‰, and dissolved oxygen content was approximately 7 mg l^− 1 during the experimental period. Survival decreased with increasing dietary MBM and the survival in the fish fed the diet with 75% protein from MBM was significantly lower than other groups (P < 0.05). There were no significant differences in specific growth rate (SGR) among the fish fed the diets with 0 (the control group), 15, 30 and 45% protein from MBM. However, SGR in the fish fed the diets with 60 and 75% protein from MBM were significantly lower than other groups (P < 0.05). No significant differences in feeding rate were observed among dietary treatments. The digestibility experiment showed that the apparent digestibility coefficients (ADC) of dry matter, protein, lipid and energy of MBM were significantly lower compared with those of FM (P < 0.05). Essential amino acid index was found to be correlated positively with SGR in the present study, suggesting that essential amino acid balance was important. Body composition analysis showed that the carcass protein and essential amino acids were not significantly affected by dietary MBM. The lipid and n-3 highly unsaturated fatty acid (n-3 HUFA) in fish muscle, however, significantly decreased with increasing dietary MBM. These results showed that 45% of FM protein could be replaced by MBM protein in diets of large yellow croaker without significantly reducing growth. It was suggested that the reduced growth with higher MBM was due to lower digestibility and imbalance of essential amino acids.     

Growth of the prawn Macrobrachium rosenbergii in asbestos asphalt ponds in hard water and on a low protein diet

Freshwater prawns Macrobrachium rosenbergii were grown for 158 days in four 25 m² ponds, two with earth bottoms and two with asbestos asphalt bottoms, in water with a total hardness of 1000 ppm and on a diet with a 14% protein content. Production was in the range of 660–1270 kg/ha with 97% of final biomass in marketable size prawns of 17–70 g. The results were comparable with other temperate region studies and suggested that water high in calcium but low in carbonates may not inhibit growth in M. rosenbergii.     

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.     

Safety and efficacy of emamectin benzoate administered in-feed to Atlantic salmon, Salmo salar L., smolts in freshwater, as a preventative treatment against infestations of sea lice, Lepeophtheirus salmonis (Krøyer)

The safety and efficacy of emamectin benzoate, administered in-feed to Atlantic salmon smolts, Salmo salar L., held in freshwater, was evaluated as a preventative treatment against sea lice, Lepeophtheirus salmonis, following transfer of fish to seawater.

In the safety study, salmon smolts held in freshwater were fed with diets containing emamectin benzoate at nominal doses of 0 (control), 50 (recommended dose) and 250 (5× recommended dose) μg kg⁻¹ fish day⁻¹ for 7 days (days 0–6). Actual dose rates, based on measured concentrations of emamectin benzoate in feed, differences in fish weight, and feed consumed, were 0, 54, and 272 μg kg⁻¹ day⁻¹, respectively. On day 9, fish were transferred to seawater and observed for 14 days. No differences in feeding response, coordination, behaviour, gross and histological appearance were observed between control fish and those that received 54 μg kg⁻¹ day⁻¹. Among smolts that received 272 μg kg⁻¹ day⁻¹, approximately 50% exhibited darker coloration, and one fish (1%) exhibited uncoordinated swimming behaviour. No pathognomonic signs of emamectin benzoate toxicity were identified.

In the efficacy study, smolts held in freshwater were fed an unmedicated ration (control group) or emamectin benzoate at 50 μg kg⁻¹ day⁻¹ (treated group) for 7 days (days 0–6). On day 9, fish were re-distributed to eight seawater tanks, each holding 30 control and 30 treated fish. On days 28, 56, 77 and 109, respectively, control and treated fish in two tanks were challenged with L. salmonis copepodites. When lice in each group reached chalimus stage IV, fish were sampled and the numbers of lice were recorded. Fish challenged at day 109 were sampled for the second time when lice were at the adult stage. Efficacy was calculated as the reduction in the mean number of lice on treated fish relative to the mean on control fish. Treatment with emamectin benzoate resulted in an efficacy of 85.0–99.8% in fish challenged at days 28–77, from the start of treatment, and lice counts were significantly lower (P<0.001) on treated fish than on controls. When fish challenged at day 109 were sampled at day 128, efficacy was 44.3%, but survival of chalimus to adult lice on treated fish was lower, and at day 159, efficacy had increased to 73%. These results demonstrate that treatment of salmon smolts with emamectin benzoate in freshwater was well tolerated and highly effective in preventing sea lice infestation following transfer of fish to seawater.