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.     

Partial replacement of fish meal by porcine meat meal in practical diets for Pacific white shrimp (Litopenaeus vannamei)

In this study, it evaluated the growth performance of the Pacific white shrimp Litopenaeus vannamei in response to the replacement of fish meal with rendered porcine meat meal (PMM) in its diet. Six isolipidic and isonitrogenous diets were formulated with 0, 25, 35, 45, 55 or 65% replacement of fish meal with PMM on a protein basis. Shrimp grew from 0.55 g to > 3.6 g during the 41-day experimental period. Specific growth rate (SGR) was significantly lower when PMM inclusion was 26.18% or greater, replacing more than 45% fish meal protein. A significant negative relationship was observed between growth response and the level of fish meal protein replacement with PMM protein. Methionine content decreased as PMM inclusion levels increased, consequently compromising growth performance. Dry feed intake (DFI) and the feed conversion ratio (FCR) were unaffected by fish meal replacement levels. The protein efficiency ratio (PER) was highest at the lowest PMM inclusion level. Apparent protein digestibility coefficient (APDC) for PPM was 66.2%. Experimental diets D-0 and D-25 had apparent dry matter digestibility (ADMDs) ranging from 77–81% and ADPs from 82–85%, while the diets with higher PMM inclusion (D-35 to D-65) had a significantly lower ADMD range (70–72%) and APD range (73–78%). It is concluded that porcine meat meal is an acceptable alternative animal protein source that can replace up to 35% of fish meal protein in shrimp diets without significant adverse effects on growth, survival, FCR, PER and body composition.     

Melanocyte-stimulating hormone facilitates hypermelanosis on the non-eyed side of the barfin flounder, a pleuronectiform fish

A pleuronectiform fish, the barfin flounder, Verasper moseri, is promising for aquaculture and resource enhancement in Northern Japan due to its high commercial value. Hypermelanosis of its non-eyed side, which frequently occurs under culture conditions, diminishes its commercial value. Two peptide hormones, melanin-concentrating hormone (MCH) and melanocyte-stimulating hormone (MSH), having opposing actions, are associated with the color changes of fish. We have previously reported the positive effect of MCH in preventing hypermelanosis. Here, we examined the effects of MSH on the occurrence of hypermelanosis. A single injection of Des-Ac--MSH [0.01 nmol/g–10 nmol/g (0.016 μg–16 μg/g)] did not change the eyed-side body color, while a single injection of MCH [0.1 nmol/g (0.21 μg/g)] made the eyed-side color paler. No difference was observed in eyed-side lightness between fish injected with MCH (0.1 nmol/g) and those receiving MCH (0.1 nmol/g) and an increased amount of Des-Ac--MSH (0.01 nmol/g–10 nmol/g) simultaneously. These results indicate that MSH does not suppress the in vivo body color-paling effects of MCH in barfin flounders. On the other hand, implantation of a cholesterol pellet containing Des-Ac--MSH (280 μg, twice at 29-day interval) increased hypermelanosis of the non-eyed side of barfin flounders compared to control fish. Eyed-side bodies of MSH-treated fish were darker than control fish; thus, MSH is involved in morphological color change including ectopic melanin synthesis in non-eyed-side skin.     

Extenders and cryoprotectants for cooling and freezing of piracanjuba (Brycon orbignyanus) semen, an endangered Brazilian teleost fish

The aim of this study was to develop a protocol for semen storage of piracanjuba (Brycon orbignyanus) by both cool storage at 4 °C and cryopreservation at − 196 °C. Semen was diluted in some fish semen extenders (Exp. 1) or in extenders combined with the antibiotic gentamycin sulfate (Exp. 2) and stored at 4 °C. Sperm motility was estimated every 24 h. Then, the effects of egg yolk (0 and 5%), cryoprotectants (dimethyl sulphoxide — DMSO, methanol, and methylglycol) and extenders (NaCl 154 mM, BTS™ Minitub and M III™ Minitub) on semen cryopreservation were evaluated (Exp. 3). Semen was added to each of eighteen cryosolutions (2 yolk concentrations × 3 cryoprotectants × 3 extenders), aspirated into 0.5-mL straws, frozen in nitrogen vapor (Taylor-Wharton, CP 300, “dry shipper”) and stored at − 196 °C. Sperm motility was evaluated after thawing at 60 °C-water bath for 8 s. The three cryosolutions that produced the highest post-thaw sperm motility were used again to freeze semen. Post-thaw semen quality was then evaluated under three tests: sperm motility, the percentage of live spermatozoa and hatching rate (Exp. 4). Piracanjuba semen diluted (1:10 total volume) in NaCl 200 mM or in Saad solution (NaCl 200 mM, Tris 30 mM) maintained motility above 35% for as long as 7 days, at 4 °C. Motility of only 7% was observed on undiluted semen after 3 days at 4 °C. There was neither beneficial nor detrimental effect of gentamycin on sperm motility at 250 μg/mL. Egg yolk addition to the cryosolution was beneficial in samples cryopreserved in NaCl 154 mM and in M III™, but detrimental for samples cryopreserved in BTS™. Methylglycol was the most effective cryoprotector compared to DMSO and methanol. Motility and percentage of live spermatozoa were similar among semen cryopreserved in NaCl–yolk, M III™–yolk and BTS™, all containing 10% methylglycol, but lower than fresh control. Hatching rates of eggs fertilized with sperm cryopreserved in NaCl–yolk or BTS™ were higher than for eggs fertilized with sperm cryopreserved in M III™–yolk, but lower than control fertilizations. The semen cryopreservation protocols developed here will be used to set up a gene bank for endangered piracanjuba populations.     

Involvement of trypsin and chymotrypsin activities in Atlantic cod (Gadus morhua) embryogenesis

Proteases play a key role in yolk formation and degradation during embryogenesis of marine fish. This study presents the first clear data on the involvement of trypsin and chymotrypsin activities in the embryogenesis of Atlantic cod (Gadus morhua). Both enzyme activities were shown to be present in unfertilized eggs followed by a significant decline (P < 0.01) in trypsin activity during the first 4 days post fertilization (dpf). Thereafter, the trypsin- and chymotrypsin activities increased to a maximum around day 9 pf. A decline in trypsin and chymotrypsin activities was observed from day 10 pf with minimal activity just prior to first feeding (day 15 pf). Western blot analysis, using polyclonal antibodies raised to Atlantic cod trypsins I and Y, mostly coincided with the trypsin activity profile. The novel trypsin Y was previously shown to have both trypsin- and chymotrypsin-like activities. Thus, some of the chymotrypsin activity observed in the samples may be originated from trypsin Y. The low trypsin and chymotrypsin activities just prior to first feeding (13–15 dpf) may indicate insufficient digestive function as trypsin has been shown to be a suitable short-term indicator reflecting the nutritional quality of marine fish larvae.     

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.     

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.     

Pharmacokinetics and the active metabolite of enrofloxacin in Chinese mitten-handed crab (Eriocheir sinensis)

The pharmacokinetics and active metabolite of enrofloxacin were estimated after single intramuscular administration (10.0 or 20.0 mg/kg body weight) to the Chinese mitten-handed crab (Eriocheir sinensis) in fresh water at 25.0 ± 1.0 °C. Levels of enrofloxacin and its metabolite ciprofloxacin in the main tissues (hemolymph, hepatopancreas, muscle, ovary and spermary) were simultaneously detected by HPLC. Enrofloxacin concentration–time profiles for the hemolymph in both tests were described by a two-compartment open model with first-order absorption. Distribution half-time (T_1/2), elimination half-time (T_1/2β), body clearance (CL/F), mean residence time (MRT_0–∞), area under the concentration–time curve from 0 to ∞ h (AUC_0–∞) and apparent volume of distribution (Vd/F), which derived from the pharmacokinetic model, were 0.427 h, 21.3 h, 0.133 l/h/kg, 60.0 h, 96.9 μg/ml/h and 4.08 l/kg, respectively, at a dose of 10.0 mg/kg body weight, and 0.216 h, 12.3 h, 0.189 l/h/kg, 85.8 h, 187 μg/ml/h and 3.35 l/kg, respectively, at a dose of 20.0 mg/kg body weight. Similarities were found between the hemolymph concentration–time curves of the two tests; for example, instant absorption process followed by the distribution phrase, and a second absorption peak at 6 h post-treatment. After intramuscular administration of 10.0 mg/kg body weight, absorption of enrofloxacin was observed in the main edible tissues (hepatopancreas, muscle, ovary and spermary), and the drug residue was the highest in the hepatopancreas, where the ‘drug sink’ phenomenon occurred. Comparative pharmacokinetics showed fast absorption, broad distribution and fast elimination of enrofloxacin in E. sinensis after intramuscular dosing. Regarding ciprofloxacin, the main active metabolite of enrofloxacin, though relatively low levels were detected in all the main tissues of the crab, its kinetics in the hemolymph in the two tests were not described by a one- or two-compartment open model.     

Effects of dietary estradiol-17β in juvenile shortnose sturgeon, Acipenser brevirostrum, Lesueur

Shortnose sturgeon, Acipenser brevirostrum, were fed estradiol-17β (E₂) in commercial salmon feed in two separate experiments, with the aim of creating all-female populations for aquaculture. In the first experiment, 5-month-old fish were fed 0 (control), 10, 50 and 100 mg E₂/kg feed during a 9-month trial starting in September. This experiment was confounded by copper contamination of the heated water source that appeared to cause E₂-dose-dependent mortality. As a result, a second, parallel experiment was conducted on the same year class of fish starting at age 7 months in November, but with hormone dosage reduced to 0, 10, 25 and 50 mg E₂/kg feed in a 7-month trial. In both experiments, control and 10 mg E₂/kg groups remained active and fed well, but the other fish were less active and fed less as E₂ dosage increased. Histological observations indicated that anatomical differentiation of the ovaries had begun in 3 of 5 individuals examined at the start of Experiment 2. In January, sub-samples of 6 fish from each group showed that E₂-treated fish had mottled livers, swollen kidneys and reduced gonad size. The number of hepatocytes and their staining characteristics were similar in both the control and 10 mg E₂/kg groups. At 25 mg E₂/kg and higher, the number of hepatocytes and the presence and quantity of yolk protein increased with the E₂ dosage. At final sampling in May, all of the treatment groups showed some individuals with these pathologies evident macroscopically, and their frequency and severity increased with E₂ dosage such that all individuals of the 100 mg E₂/kg group were affected. In general, fish grew better and had lower mortality in Experiment 2 (8.5%, as compared to 15.2% for Experiment 1) during the period of E₂ treatment. Final sampling revealed that all treated fish were apparently feminized in both experiments, based on anatomical differentiation of the gonads. Control groups, on the other hand, were 65 and 58% female in Experiments 1 and 2, respectively.     

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.     

Nutrients removed by Kappaphycus alvarezii (Rhodophyta, Solieriaceae) in integrated cultivation with fishes in re-circulating water

The potential of the red alga Kappaphycus alvarezii to remove nutrients was tested to treat effluents of Trachinotus carolinus fish cultivation, and the production of carrageenan in this condition was analyzed. Experiments were conducted in four tanks of 8000 L with approximately 1200 fishes of 30 g each integrated with three tanks of 100 L with 700 g of K. alvarezii, as initial biomass per tank. Seawater was re-circulated between tanks with seaweed and with fish. As a control, three tanks with seawater circulating in an open system were utilized. Seawater samples were collected daily for 10 days and concentrations of nitrate, nitrite, ammonium and phosphate were determined in the inflow and outflow water of the tanks. Significant differences between both collecting points were considered as nutrient removal by the seaweed. Growth rates and carrageenan yields were also analyzed in seaweed cultivated in seawater and in effluents. Growth rates of seaweed cultivated in tanks were lower than those obtained in open sea and in laboratory cultivation. Effluents had concentrations of nitrate and nitrite ca. 100 times higher than in the control. Maximum values of nutrient removal on effluents were: nitrate = 18.2%; nitrite = 50.8%; ammonium = 70.5% and phosphate = 26.8%. All plants survived throughout the experimental period, but some developed “ice–ice”, a disease associated with physiological stress. After the experimental period, some plants selected and cultivated in open sea presented higher growth rates in 40 days, indicating nutrient storage. No significant differences between carrageenan yields of K. alvarezii cultivated in seawater and in the effluents were observed. Our results show that K. alvarezii can be utilized as a biofilter for fish cultivation effluents, reducing the eutrophication process and can also be processed for carrageenan production, which provides an additional benefit to the fisheries.     

Regulatory costs on U.S. salmonid farms

The economic effects of the implementation of regulations on aquaculture farms in the United States, while of concern, are not well understood. A national survey was conducted of salmonid (trout and salmon) farms in 17 states of the United States to measure on‐farm regulatory costs and to identify which regulations were the most costly to this industry segment. The response rate was 63%, with a coverage rate of 94.5% of the U.S. production of salmonids. The regulatory system resulted in increased national on‐farm costs of $16.1 million/year, lost markets with a sales value of $7.1 million/year, lost production of $5.3 million/year, and thwarted expansion attempts estimated at $40.1 million/year. Mean farm regulatory costs were $150,506/farm annually, or $2.71/kg; lost markets with annual sales values of $66,274/farm; annual lost production of $49,064/farm; and an annual value of thwarted expansion attempts estimated at $375,459/farm. Smaller‐scale farms were affected to a disproportionately greater negative extent than larger‐scale farms. Per‐farm regulatory costs were, on average, greater for foodfish producers than for producers selling to recreational markets, but per‐kg regulatory costs were greater for those selling to recreational compared to foodfish markets. Regulatory costs constituted 12% of total production and marketing costs on U.S. salmonid farms. The greatest regulatory costs were found to be effluent discharge regulations. The majority of regulatory costs were fixed costs, but regulatory barriers to expansion precluded compensatory adjustments to the business in spite of growing demand for salmonid products. Results of this study show that the on‐farm regulatory cost burden is substantial and has negatively affected the U.S. salmonid industry's ability to respond to strong demand for U.S. farm‐raised salmonid products. Results also suggest that the regulatory system has contributed to the decline in the number of U.S. salmonid farms. While regulations will necessarily have some degree of cost to farms, the magnitude of the on‐farm regulatory cost burden on U.S. salmonid farms calls for concerted efforts to identify and implement innovative regulatory monitoring and compliance frameworks that reduce the on‐farm regulatory cost burden.     

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.     

Efficacy, tissue distribution, and residue depletion of oxytetracycline in WS-RLP infected California red abalone Haliotis rufescens

Oxytetracycline (OTC) was recently demonstrated to be an effective therapeutant for California red abalone (Haliotis rufescens) infected with the agent of withering syndrome (WS), a Rickettsia-like prokaryote (WS-RLP). This study examined the influence of temperature on the efficacy and pharmacokinetics of oral OTC treatments on WS-RLP infected red abalone, H. rufescens. Medication was administered in an artificial diet containing 1.85% active OTC at a rate of 103.4 mg/kg abalone for 10, 20 and 30 days at both 13.4 °C and 17.3 °C. Drug residue concentrations, WS-RLP burden and WS-associated pathological changes within foot muscle and digestive gland were measured 3, 17, 23, 42, 63, 81, 102, 122, and 160 days after OTC treatment. In all samples the highest concentrations of OTC were detected on the first sample day. Furthermore, all sampling periods showed significantly less OTC in foot muscle samples relative to corresponding digestive gland samples (p < 0.001). While drug depuration occurred rapidly from foot muscle, unusually high concentrations and long depuration times were measured within the digestive gland. The pharmacokinetic trends in both the foot muscle and digestive gland at 13.4 °C were adequately described by a one-compartment model, while the digestive gland depuration models within the 17.3 °C treatments conformed to a two-compartment model. At both temperatures treatment duration gave rise to significant differences in OTC accumulation and depletion (p < 0.001). Despite this, terminal elimination rate constants were similar. Estimated half-lives were similar among all treatments for digestive gland (23.8–27.5 d) and all treatments for the foot muscle at 13.4 °C (10.7–13.4 d; calculated only for the 13.4 °C treatments). At both temperatures all three durations of drug treatment led to significant reductions in WS-RLP prevalence, foot muscle atrophy, and mortality. WS-RLP re-challenge trials initiated 44, 88, and 122 days following the 10-day treatment at 17.3 °C showed reduced susceptibility to re-infection for up to 88 days after treatment, suggesting that digestive gland OTC residues confer long-term resistance to pathogen. Atomic absorption spectroscopy conducted on both foot muscle and digestive gland found significantly higher concentrations of iron, zinc, and manganese in the digestive gland, offering an explanation behind the underlying mechanism of OTC retention for extended periods within this tissue.     

Temperature effects on early development and occurrence of metamorphosis-related morphological abnormalities in hatchery-reared brown sole Pseudopleuronectes herzensteini

The objectives of the present study are to elucidate the effects of rearing temperatures on early larval development and the occurrence of metamorphosis-related morphological abnormalities in hatchery-reared brown sole Pseudopleuronectes herzensteini. Newly hatched larvae were reared through metamorphosis at different temperatures (6, 9, 12, 15, 18, 21, and 24 °C). Growth and development of larvae were accelerated concomitant with higher temperatures. Metamorphosed juveniles were classified into four morphological categories as normal, pseudoalbinism (two types), and ambicoloration, in combination with body color (pigmentation) and eye location. Incidence of normal morphology was lowest (11–24%) and that of pseudoalbinism was highest (74–83%) at 12 °C. The highest incidence of normal morphology (51–73%) was obtained at 21 °C. Although occurrence of ambicoloration was very low in all temperature regimes (0–10%), it increased slightly with increasing temperatures. Results of the present study strongly suggested that rearing temperature during the larval period affected the appearance of normal fish because of the different larval growth and development.     

Thyroid gland development in Senegalese sole (Solea senegalensis Kaup 1858) during early life stages: A histochemical and immunohistochemical approach

A key to success in the culture of marine fish species is the mass production of high quality fry, a process largely dependent on successful first feeding and normal development and growth of fish larvae. In this regard it is important to examine the structural and functional development of the endocrine system (pituitary, thyroid, interrenal glands) during early ontogeny of marine fish. Particularly, the thyroid hormones, thyroxine (T₄) and triiodothyronine (T₃), influence numerous metabolic processes, such as growth, differentiation, metamorphosis, reproduction, respiration, migratory behaviour, central nervous system activity, seasonal adaptation, etc. Therefore the aim of this study was to describe the development of the thyroid gland and the ontogeny appearance of the thyroid hormones in Solea senegalensis larvae by means of histological and immunohistochemical techniques. The first thyroid follicle was present at 4 days-post-hatch (dph) coinciding with first feeding. During metamorphosis (12–20 dph) the follicles increased in both number and size, and by 30 dph presented the same characteristics as that seen in adult fish. Tissue immunostaining of both thyroid hormones decreased during the endogenous larvae development to nearly undetectable levels at the completion of yolk-sac absorption. During larvae exogenous phase, T₃ and T₄ immunostaining was first detected by 6 dph and an increase of specific staining for both hormones was detected between 12 and 20 dph, during metamorphosis phase.     

Long photoperiod delayed spawning and increased somatic growth in gilthead seabream (Sparus aurata)

The effects of extended photoperiods, mimicking the longest day of the year, were studied in 1- and 2-year seabream. The photoperiod regimes started in late July, 36 and 39 days after the summer solstice and continued for 11 months, well beyond the natural reproductive season of December–March. Regime 1 (long day, 15.5L:8.5D), which used natural and fluorescent light, reduced the incidence of maturity in both year classes and females did not spawn although some gonadal development was observed. Among all 1-year sampled fish of regime 1, a maximum of 5% became spermiating males (March) and 5% reached the yolk granule stage of vitellogenesis (VO3; 250–400 μm diameter) by May. Among 2-year sampled fish of regime 1, 45% became spermiating males and 25% were females, which reached the advanced vitellogenesis stage (VO4; 400–600 μm) by April. Regime 2 (skeleton photoperiod), consisting of natural light and a 1.5-h pulse of fluorescent light during the period 14–15.5 h after sunrise, postponed gonadal development and spawning for up to 3 months. In this regime, a maximum of 80% of 1-year sampled fish were spermiating males in February and a maximum of 10% were VO3 stage females in March. In the sampled 2-year fish, the maximum levels were 50% spermiating males in February and 25% VO3 stage females in March. Control fish, which were exposed to the natural photoperiod (29°34′N), spawned during their natural season. The maximum levels for 1-year sampled control fish were 95% spermiating males and no females in December, while among 2-year sampled fish, maxima of 75% males in February and 45% VO4 stage females in November. Final average weights of photoperiod treated fish (1-year=430 g—regime 1, 400 g—regime 2; 2-year=582 g—regime 1, 518 g—regime 2) were significantly greater (p<0.05) than control fish (1-year=341 g; 2-year=476 g). Daily feed consumption (g/100 g fish) dropped from an average of 1.83 to 0.93 g for 1-year fish during August–December and from 0.88 to 0.54 g for 2-year fish during the same period. This was correlated with reduced autumn and winter water temperatures (26–20°C summer to winter change) and increased fish weight in all treatments. Daily feed consumption was also affected by the onset of spawning in the control and regime 2 (skeleton photoperiod) treatments of both 1- and 2-year fish. The efficiency of feed utilization (FCR) and protein and energy retention were all affected by the photoperiod regimes and followed the same pattern of decrease as weight gain, regime 1 (long day)>regime 2 (skeleton photoperiod)>control.     

Swimming stimulates oocyte development in European eel

In this study, we subjected eels from Lake Balaton (Hungary) to a swimming period of 1 week and 2 or 6 weeks. Most eels were silver and were 13–21 years old. Time dependent changes in morphometrical parameters and developmental characteristics of the oocytes were determined. Already after 1 week of swimming, the gonadal mass increased and oocytes became larger, filled with large numbers of lipid droplets. After 2 and 6 weeks of swimming we found in addition a significant enlargement of the eyes, which is a sign of sexual maturation. In contrast to the resting eels, that had oocytes in the primary growth phase (stage 1–2); the swimming eels had oocytes in stage 3; the cortical alveolus or lipid droplet stage. The results indicate that lipid mobilisation induced by swimming is a requirement for the natural incorporation of lipid droplets in the oocytes, a crucial step in oocyte maturation. As the Balaton eels responded stronger to swimming than young farmed eels, it is suggested that older eels are more sensitive for maturation triggers.     

The effects of salinity on the survival, growth and haemolymph osmolality of early juvenile blue swimmer crabs, Portunus pelagicus

The blue swimmer crab, Portunus pelagicus, is an emerging aquaculture species in the Indo-Pacific. Two experiments were performed to determine the effects of salinity on survival, growth and haemolymph osmolality of early juvenile P. pelagicus crabs. The salinities tested for the first experiment were 10, 15, 25 and 40 ppt, and for the second experiment 5, 20, 30, 35 and 45 ppt. Each salinity experiment was triplicated, with each replicate consisting of 10 stage 4 juveniles. Each experiment lasted 45 days. Mortalities and incidence of “molt death syndrome” were recorded daily, while the intermolt period, carapace length, carapace width and wet weight were measured at each molt. At the end of the experiments the haemolymph osmolality and dry weights were measured.

Results demonstrate that salinity significantly affects both the survival and growth of early P. pelagicus juveniles. Mortality was significantly higher (p < 0.01) for juveniles cultured at salinities ≤ 15 ppt and at 45 ppt. At a salinity of 5 ppt a complete mortality occurred on day 20. In all salinity treatments, the majority of mortalities were due to “molt death syndrome”. In experiment 1, immediate effects of salinity on growth and development were seen at 10 ppt as the intermolt period was significantly longer (p < 0.01) and the mean carapace size increase was significantly less (p < 0.01) at the first molt compared to the other treatments. Meanwhile, the specific growth rates (carapace length, width and wet weight) were significantly lower (p < 0.05) at high salinities (≥ 40 ppt) due to longer intermolt periods and significantly lower (p < 0.05) carapace size or wet weight increases.

The haemolymph osmolality exhibited a positive linear relationship with the culture medium with an isosmotic point of 1106 mOsm/kg, equal to a salinity of approximately 38 ppt. Based on the osmolality graph, high metabolic cost for osmoregulation due to increased hyper- and hypo-osmotic stress appeared to cause lower survival and specific growth rates of the crabs. The results demonstrate that a salinity range of 20–35 ppt is suitable for the culture of early juvenile P. pelagicus.     

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.