Calculation of pH in fresh and sea water aquaculture systems

A procedure for the calculation of pH in fresh and salt waters has been developed. The method is based on a fourth-order polynomial relationship between hydrogen ion concentration and other (conservative) water quality parameters. The method avoids trial and error estimations and results in a direct calculation procedure that can be implemented in models developed in various modeling environments, such as spreadsheets, conventional programming languages (BASIC, C, FORTRAN, PASCAL, etc.) or specialized modeling languages (Extend^™, Stella^™).

The method developed is based on the solution of the full alkalinity-pH equation. Because of the need for simplification of the equations to yield explicitly solvable polynomial equations, the accuracy of the solutions depends on the simplification made and varies with water properties. Three simplifications are tested based on a second-, a third-and a fourth-order polynomial equation for hydrogen ion concentrations. the equations have been tested for salinities ranging from 0 to 35‰ (fresh to sea water), for temperatures ranging from 0 to 35°C, for total carbonate carbon concentrations of 0·1 and 5·0 mmol/liter, and for total ammonia nitrogen concentrations of 0 and 10 mg/liter. Approximations are most accurate in waters of high total carbonate carbon and low ammonia concentrations, where the fourth-order approximation yields results that are within 0.05 pH units for the full range of pH values tested (5–10).     

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.     

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.     

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.     

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.     

Characterization of Ca-ATPase activity in gill microsomes of freshwater mussel, Lamellidens marginalis (Lamarck) and heavy metal modulations

Freshwater mussel Lamellidens marginalis showed a Ca²⁺-stimulated ATPase activity in the microsomal fraction of gill. Mg²⁺ was also found to be an activating cation for this ATPase enzyme. Ca²⁺ ATPase showed maximal activity at 40 °C and between pH 7.5–8.0. Substrate specificity of Ca²⁺ ATPase was highest with ATP, followed by GTP and other trinucleotides such as UTP, CTP and ADP also showed some amount of hydrolysis. Ca²⁺ ATPase showed slight inhibition with ruthenium red and sodium vanadate and is insensitive to sodium azide, ouabain, oligomycin B and phenylalanine. Heavy metals like Hg²⁺, Cu²⁺ and Zn²⁺ showed 50% inhibition of Ca²⁺ ATPase activity at concentrations of 0.25 mM, 0.5 mM and 1 mM, respectively whereas Cd²⁺ and Ni²⁺ showed 39% and 28% inhibition of enzyme activity at 1 mM concentrations.     

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.     

Energy balance of Litopenaeus vannamei postlarvae fed on animal or vegetable protein based compounded feeds

L. vannamei postlarvae are normally raised with a protein dense diet (50% protein) rich in fishmeal. Part of the protein is utilized for energy purpose instead of protein synthesis. Based on a previous energy partitioning study, the effects of two isoenergetic compounded feed treatments – animal protein (AP) and vegetable protein and carbohydrates (VPC) – upon growth efficiency and energy budget of shrimp postlarvae and early juveniles were determined. Recovered energy (RE) or production (P) after 50 days trial was similar (2 J day^− 1) in both treatments, from PL₁₄ to PL₁₉. However, early juveniles discriminated between animal protein (116 J day^− 1) and vegetable protein and carbohydrates (88 J day^− 1). The difference in respiration indicated a higher heat increment with AP compared to VPC. At maintenance level, energy used was lower with AP than VPC treatment. Postlarvae and early juveniles employed protein as a main energy substrate (O:N < 20). Differences in the efficiencies observed in the calculated energy budget were attributed to the presence of carbohydrates in diet and not to the protein source. The advantage of incorporating vegetable protein source in the diet of harvesting shrimp may eventually contribute towards a reduction of fishmeal costs and waste products as well as to achieve sustainable shrimp farming.     

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.     

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.     

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.     

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.     

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.     

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.     

Turbidity related to shrimp size in tanks with soil substrate

Water turbidity was monitored daily during a 63-day growth trial with juvenile Penaeus vannamei in indoor laboratory tanks. The tanks contained a 2.5 cm layer of soil substrate collected from 14 ponds representing five shrimp farms in Texas. A linear regression model relating turbidity to estimated shrimp length accounted for 49–83% of the total variation in turbidity for the 14 soil groups. For averages of daily turbidity and shrimp length over the 14 soil groups, the linear regression model yielded an r² = 0.92. Turbidity increased with shrimp length (75–135 mm), apparently because the elevated locomotory or feeding activity of the larger shrimp increased the suspension of soil particles. These results suggest the potential of using water turbidity as a tool for estimating shrimp biomass in ponds.     

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.     

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.     

Manipulation of ploidy in yellow perch (Perca flavescens) by heat shock, hydrostatic pressure shock, and spermatozoa inactivation

Heat shocks, hydrostatic pressure shocks, and ultraviolet radiation were evaluated for their efficacy as methods of manipulating ploidy in yellow perch (Perca flavescens). The most effective methods of inducing triploidy were heat shocks of 28–30°C applied at a time of initiation (TI) of 5 min postfertilization for durations of 10 or 25 min, and hydrostatic pressure shocks of 9000 or 11 000 psi applied at a TI of 5 min for a duration of 12 min. These treatments resulted in triploidy induction rates that ranged from 54–100%, and embryonic survival rates of 16–80%. Cold shocks of 0°C had no effect on the ploidy or survival of embryos. For perch, hydrostatic pressure shock offered several advantages over heat shock as a method of manipulating ploidy. The most effective methods of inducing tetraploidy were hydrostatic pressure shocks of 9000 psi applied at a TI of 192 min for durations of 16 or 24 min. Ultraviolet radiation of perch sperm with doses of 3240–6480 ergs/mm² resulted in 100% inactivation of paternal chromosomes, and perch eggs fertilized with inactivated sperm had survival rates of > 50%, thereby establishing methods for producing gynogenetic perch. Studies comparing the growth and performance of diploid vs. triploid perch are underway. Tetraploid perch are being reared to sexual maturity to evaluate their potential as brood fish.     

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.