Purification and characterization of α<Subscript>1</Subscript>-proteinase inhibitor and antithrombin III: major serpins of rainbow trout (<Emphasis Type="Italic">Oncorhynchuss mykiss</Emphasis>) and carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis>) blood plasma

The main serine proteinase inhibitors of rainbow trout (Oncorhynchuss mykiss) and common carp (Cyprinus carpio) blood plasma were isolated and purified. The investigated inhibitors, α₁-proteinase inhibitor (α₁-PI) and antithrombin III (AT III), act by forming stable complexes with target proteinases. The association rate constants k _on for the interaction of fish plasma inhibitors with several serine proteinases have been determined: k _on for both carp and rainbow trout α₁-PI were >10⁷ M⁻¹ s⁻¹ for human neutrophil elastase, and in the case of bovine trypsin and chymotrypsin k _on values were 2.0–5.2 × 10⁶ M⁻¹ s⁻¹. Association rate constants k _on for the interaction of carp and rainbow trout AT III with bovine trypsin and thrombin were about 1.3 × 10⁴–7.9 × 10⁵ M⁻¹ s⁻¹ without and >10⁷ M⁻¹ s⁻¹ in presence of heparin; so antithrombins require the presence of heparin to become effective proteinase inhibitors. The high degree of homology of the estimated amino acid sequences of fish inhibitors reactive site loops confirms their similarity with other proteinase inhibitors from the serpin family.     

Purification and characterization of pepsinogens and pepsins from the stomach of rice field eel (<Emphasis Type="Italic">Monopterus albus</Emphasis> Zuiew)

Three pepsinogens (PG1, PG2, and PG3) were highly purified from the stomach of freshwater fish rice field eel (Monopterus albus Zuiew) by ammonium sulfate fractionation and chromatographies on DEAE-Sephacel, Sephacryl S-200 HR. The molecular masses of the three purified PGs were all estimated as 36 kDa using SDS–PAGE. Two-dimensional gel electrophoresis (2D-PAGE) showed that pI values of the three PGs were 5.1, 4.8, and 4.6, respectively. All the PGs converted into corresponding pepsins quickly at pH 2.0, and their activities could be specifically inhibited by aspartic proteinase inhibitor pepstatin A. Optimum pH and temperature of the enzymes for hydrolyzing hemoglobin were 3.0–3.5 and 40–45°C. The K _m values of them were 1.2 × 10⁻⁴ M, 8.7 × 10⁻⁵ M, and 6.9 × 10⁻⁵ M, respectively. The turnover numbers (k _cat) of them were 23.2, 24.0, and 42.6 s⁻¹. Purified pepsins were effective in the degradation of fish muscular proteins, suggesting their digestive functions physiologically.     

Delivery of HUFA, probionts and biomedicine through bioencapsulated Artemia as a means to enhance the growth and survival and reduce the pathogenesity in shrimp Penaeus monodon postlarvae

One of the major problems in the shrimp culture industry is the difficulty in producing high-quality shrimp larvae. In larviculture, quality feeds containing a high content of highly unsaturated fatty acids (HUFA) and ingredients that stimulate stress and disease resistance are essential to produce healthy shrimp larvae. In the present study, Penaeus monodon postlarvae (PL15) were fed for 25 days on an unenriched Artemia diet (control; A) or on a diet of Artemia enriched with either HUFA-rich liver oil of the trash fish Odonus niger (B), probionts [Lactobacillus acidophilus (C1) or yeast-Saccharomyces cerevisiae (C2)] or biomedicinal herbal products (D) that have anti-stress, growth-promoting and anti-microbial characteristics. P. monodon postlarvae fed unenriched Artemia exhibited the lowest weight gain (227.9 ± 8.30 mg) and specific growth rate (9.95 ± 0.05%), while those fed the HUFA-enriched Artemia (B) exhibited the highest weight gain and specific growth rate (362.34 ± 12.56 mg and 11.77 ± 0.08%, respectively). At the end of the 25-day rearing experiment, the shrimp postlarvae (PL40) were subjected to a salinity stress study. At both low and high (0 and 50‰) salinities, the group fed the control diet (A) experienced the highest cumulative mortality indices (CMI) 935.7 ± 2.1 and 1270.7 ± 3.1, respectively. Those fed diet D showed the lowest stress-induced mortality, and CMI were reduced by 31.1 and 32.3% under conditions of low and high salinity stress, respectively. A 10-day disease challenge test was conducted with the P. monodon postlarvae (PL40–PL50) by inoculating the shrimp with the pathogen Vibrio harveyi at the rate of 10⁵–10⁷ CFU/ml in all rearing tanks. P. monodon postlarvae fed probiont-encapsulated Artemia diets (C1 and C2) exhibited the highest survival (94.3 and 82.3%, respectively) and lowest pathogen load (V. harveyi) in hepatopancreas (5.2 × 10² ± 9.0 × 10 and 4.6 × 10² ± 9.0 × 10 CFU g⁻¹, respectively) and muscle (2.0 × 10² ± 6 × 10 and 1.7 × 10² ± 8.6 × 10 CFU g⁻¹, respectively) tissues. The shrimp that were fed the unenriched Artemia (Control; A) showed the lowest survival (26.33%) and highest bacterial load in the hepatopancreas (1.0 × 10⁵ ± 5 × 10³ CFU g⁻¹) and muscle (3.6 × 10⁴ ± 6 × 10² CFU g⁻¹). The shrimp fed the herbal product (D)-enriched Artemia also exhibited enhanced survival and reduced V. harveyi load in the tissues tested compared to the control diet (A) group. The results are discussed in terms of developing a quality larval feed to produce healthy shrimp larvae.     

Difference Between Tetrodotoxin and Saxitoxins in Accumulation in Puffer Fish Takifugu rubripes Liver Tissue Slices

In vitro accumulation of tetrodotoxin (TTX) and paralytic shellfish toxin (PST) in tiger puffer fish Takifugu rubripes was investigated using liver tissue slices. When T. rubripes liver slices were incubated with Leibovitz’s L-15 medium containing 0.13 mM TTX at 20 °C in air with saturated humidity, they accumulated 21.5 ± 7.3 μg TTX g⁻¹ liver after the incubation for 12 h and increased to 55.3 ± 8.2 μg TTX g⁻¹ liver at 48 h. In the incubation of T. rubripes liver slices with 0.13 mM PST-containing medium, PST was detected 6.3 ± 0.9 μg g⁻¹ liver at 12 h and reached a plateau thereafter. These results reveal the difference between TTX and PST in accumulation in T. rubripes liver tissue slices. To examine the variation in PST accumulation among fish species, the liver tissue slices from tiger puffer fish T. rubripes, parrot-bass Oplegnathus fasciatus and green ling Hexagrammos otakii were incubated at a concentration of 0.027 mM PST. The toxin contents of 3.0 μg g⁻¹ liver were observed at 8 h regardless of fish species but were not increased subsequently, showing no variety among these three species as to accumulation patterns of PST. It is noted that the tiger puffer fish T. rubripes liver specifically accumulate TTX in preference to PST.     

Effects of different natural or prepared diets on growth and survival of juvenile spider crabs, <Emphasis Type="Italic">Maja brachydactyla</Emphasis> (Balss, 1922)

The effects of different diets (natural or pellets) on growth, survival, and moulting interval of juvenile spider crabs, weighing between 0.011–1.56 g and up to 17.6 mm in carapace length, were tested over a period of 90 days. During experiment I, five diets were tested: (1) frozen shrimp—Paleomonetes sp., (2) fresh mussels—Mytilus sp., (3) white fish fillets—Merlucius merlucius, (4) blue fish fillets—Sardina pilchardus, and (5) commercial crustacean pellets. Spider crabs fed fresh mussels grew larger (0.98 ± 0.69 g) and had higher growth rates (4.0 ± 0.7 %BWd⁻¹) compared to the other four diets. The crabs fed shrimp pellet and frozen shrimp grew to intermediate sizes and were smaller than the ones fed fresh mussels, but they were larger than spider crabs fed either blue or white fish fillets (0.46 ± 0.63 and 0.26 ± 0.13 g, respectively) compared to the ones fed white fish fillets (0.12 ± 0.04) and blue fish fillets (0.04 ± 0.02 g). The spider crabs fed blue fish fillets only lasted until day 60 of the experiment, after this day none of the 20 fed this diet were left. During experiment II, two diets were tested: (1) white and blue fish fillets and (2) commercial fish pellet. There were no differences in growth both in weight or carapace length (2.9 ± 1.8 and 2.1 ± 1.5 g in weight, and 18.9 ± 5.0 and 17.7 ± 3.3 mm, respectively) at the end of the experiment. Similarly, there were no differences in growth rates in weight between the two diets (1.2 ± 0.4 and 0.9 ± 0.3 %BWd⁻¹, respectively) or in carapace length (0.4 ± 0.1 and 0.4 ± 0.2 %BWd⁻¹, respectively). Fresh mussel appears to be a very good diet to culture the early stages of this species, while shrimp pellets also deliver acceptable results. On the contrary, frozen shrimp, fish fillets either from blue or white species (much higher lipid content in the blue species), and fish pellets were found to be bad diets for the culture of the early stages of M. brachydactyla.     

The impact of net-isolated polyculture of tilapia (<Emphasis Type="Italic">Oreochromis niloticus</Emphasis>) on plankton community in saline–alkaline pond of shrimp (<Emphasis Type="Italic">Penaeus vannamei</Emphasis>)

We used 12 land-based experimental enclosures (6 m × 5 m) in a saline–alkaline pond of shrimp (Penaeus vannamei) to determine the impact of net-isolated polyculture of tilapia (Oreochromis niloticus) on plankton communities for 40 days. Tilapias were stocked in net cages suspended in enclosures, in polyculture ponds including tilapia and shrimp. Four tilapia biomass were tested: 0, 39, 115 and 227 g m⁻². Shrimp stocking biomass were 0.7 g m⁻² in all treatments. There were three replicates in each treatment. Our results showed that the presence of tilapia significantly reduced phytoplankton biomass directly through predation and indirectly through top-down effect. The stocking of tilapia reduced zooplankton biomass, particularly rotifer biomass. However, copepod biomass was not been significantly affected. So, net-isolated polyculture of tilapia can thus have a strong impact on phytoplankton allowing the co-existence of large numbers of copepods with planktivorous fish and improving the water quality of shrimp ponds.     

Growth of giant tiger prawn, <Emphasis Type="Italic">Penaeus monodon</Emphasis> Fabricius,under co-culture with a discarded filamentous seaweed, <Emphasis Type="Italic">Chaetomorpha ligustica</Emphasis> (Kützing) Kützing,at an aquarium-scale

Growth of juvenile giant tiger prawn, Penaeus monodon Fabricius, was evaluated at an aquarium-scale in co-culture with a discarded filamentous seaweed, Chaetomorpha ligustica (Kützing) Kützing. Juveniles at different ages in days were examined, designated as J 16, J 44, J 58, J 93 and J 128, where a 1-day-old juvenile (J 1) is equivalent to a 20-day-old post-larva (PL 20)). Juveniles at every age group grazed directly on live C. ligustica, even those fed an artificial shrimp diet to satiation. Mean specific growth rate (SGR: % day⁻¹) was higher in early age juveniles. Compared to mono-culture, significant differences in growth were observed at J 16 (4.44% day⁻¹) and J 44 (1.60% day⁻¹); however, no significant differences were recorded at J 58 (1.16% day⁻¹), J 93 (0.75% day⁻¹) or J 128 (0.45% day⁻¹). It was concluded that co-culture of giant tiger prawn with C. ligustica has a dietary advantage, especially in early age juveniles.     

Effects of temperature and salinity on oxygen consumption and ammonia excretion of juvenile miiuy croaker, <Emphasis Type="Italic">Miichthys miiuy</Emphasis> (Basilewsky)

The metabolic responses of the juvenile Miichthys miiuy in terms of oxygen consumption and ammonia excretion to changes in temperature (6–25°C) and salinity (16–31 ppt) were investigated. At a constant salinity of 26 ppt, the oxygen consumption rate (OCR) of the fish increased with an increase in temperature and ranged between 133.38 and 594.96 μg O₂ h⁻¹ g⁻¹ DW. The effect of temperature on OCR was significant (P < 0.01). Q₁₀ coefficients were 6.80, 1.41, 1.29 and 2.36 at temperatures of 6–10, 10–15, 15–20 and 20–25°C, respectively, suggesting that the juveniles of M. miiuy will be well adapted to the field temperature in the summer, but not in the winter. The ammonium excretion rates (AER) of the fish were also affected significantly by temperature (P < 0.01). The O:N ratio at temperatures of 6, 10, 15 and 20°C ranged from 13.12 to 20.91, which was indicative of a protein-dominated metabolism, whereas the O:N at a temperature of 25°C was 51.37, suggesting that protein-lipids were used as an energy substrate. At a constant temperature of 15°C, the OCRs of the fish ranged between 334.14 (at 31 ppt) and 409.68 (at 16 ppt) μg O₂ h⁻¹ g⁻¹ DW. No significant differences were observed in the OCR and AER of the juveniles between salinities of 26 and 31 ppt (P > 0.05). The OCR and AER at 16 ppt were, however, significantly higher than those at 26 and 31 ppt (P < 0.05), indicating salinity lower than 16 ppt is presumably stressful to M. miiuy juveniles.     

Growth parameters of whiteleg shrimp Litopenaeus vannamei and red seaweed Gracilaria corticata in integrated culturing method under zero water exchange system

Growth parameters of whiteleg shrimp Litopenaeus vannamei and red seaweed Gracilaria corticata were measured using integrated culturing method under zero‐water exchange system in a 45‐day period. A 2 × 3 factorial design was used with two levels of shrimp stocking densities and three levels of seaweed weight densities. G. corticata was cultured on a net tied to a round polyethylene frame. Culture tanks were filled with 750‐L filtered seawater. A 40‐W compact fluorescent lamp was hung over each tank to provide adequate and sufficient light for seaweed growth. Growth parameters of shrimp and seaweed such as specific growth rate (SGR), weight gained (WG) and average daily growth (ADG) were computed based on the initial and final weight of shrimp and seaweed. The maximum and minimum SGR of L. vannamei (1.97 and 1.69%/day) were observed in treatment S₁A₃ (25 shrimp/m² and 400 g seaweed/m²) and S₂A₁ (50 shrimp/m² without seaweed) respectively. The best survival rate (94.67 ± 1.33%), WG (129.9 ± 2.9%) and feed conversion ratio (1.67 ± 0.04) were also observed in treatment S₁A₃. The SGR of G.corticata in the treatment S₁A₃ (1.97 ± 0.00%/day) was significantly higher, compared to others. Strong positive correlations were obtained between the density of G. corticata and the growth parameters of L. vannamei. The red seaweed G. corticata could boost the growth parameters, survival rate and total production of L. vannamei under zero‐water exchange system.     

Acute toxicity of ammonia on juvenile cobia (Rachycentron canadum, Linnaeus, 1766) according to the salinity

Juvenile cobia (Rachycentron canadum) (total length 15.16 ± 0.92 cm and weight 19.26 ± 4.5 g) were exposed to different concentrations of ammonia–N (unionized plus ionized ammonia as nitrogen), using the static renewal method at different salinity levels of 5, 20, and 35‰ at pH 8.1 and 25°C. The 24, 48, 72, 96 h LC₅₀ values of ammonia–N for R. canadum juveniles were 60.28, 48.57, 37.42, 22.73 mg l⁻¹ at 35‰; 51.25, 43.63, 28.17, 19.05 mg l⁻¹ at 20‰; and 39.48, 25.31, 19.50, 8.13 mg l⁻¹ at 5‰, respectively. The 24, 48, 72, 96 h LC₅₀ values of NH₃–N (unionized ammonia as nitrogen) were 1.81, 1.46, 1.12, and 0.68 mg l⁻¹ at 35‰; 1.75, 1.49, 0.96, and 0.65 mg l⁻¹ at 20‰; and 1.52, 0.97, 0.71, and 0.31 mg l⁻¹ at 5‰, respectively. As the salinity decreased from 35 to 5‰, susceptibility of ammonia–N increased by 34.5, 47.88, 50.56, and 64.23% after 24, 48, 72, and 96 h exposure, respectively. Furthermore, we found that exposure of fish to ammonia–N caused an increase in oxygen consumption of 129.1, 157.5, and 192% and a decrease in the ammonia excretion level of 53.4, 38.2, and 23.3% with respect to the control.     

Age and growth of Gerres sp. (Perciformes: Gerreidae) in Okinawa Island of Southern Japan

A total of 518 Gerres sp. were collected around Okinawa Island, Japan, from November 2002 to July 2005, with monthly sampling where the standard length of females (n=218) were 56.2–147.1 mm, and males (n=149) were 62.2–139.4 mm. The maximum ages observed for females were 5⁺ years and males were 4⁺ years, estimated by transverse sectioned sagittal otoliths. Mean marginal increment indicated that opaque rings were formed once a year during April to July. The standard length (SL; mm) — body wet weight (BW; g) relationships were described as BW=(3.26×10⁻⁵) SL^2.97 and BW=(3.13×10⁻⁵) SL ^2.98 for females and males, respectively, and the standard length at age described by von Bertalanffy growth function for females, L _t=137.1(1−e^{−0.80[t+0.80]}) and males, L _t=127.3(1−e^{−0.82[t+0.93]}).     

Ex situ and in situ measurements of juvenile yellowfin tuna <Emphasis Type="Italic">Thunnus albacares</Emphasis> target strength

To provide target strength (TS) information for estimating the body length of yellowfin tuna Thunnus albacares and its abundance around fish aggregating devices, TS was measured ex situ and in situ. In the ex situ TS measurements, two cameras synchronized with a 200 kHz echosounder were used to obtain the precise orientation of the yellowfin tuna under free swimming conditions. The ex situ TS (dB re 1 m²)–fork length (FL, cm) regression was: TS = 27.06 log (FL) − 85.04. Ex situ TS was found to reach its maximum in the tilt angle range of −15° to −20° after excluding TS samples with insignificant correlation to the tilt angle. The angle between the vertebra and the swim bladder was approximately 25° according to X-ray images, supporting the above tilt range. The relationship between the swim bladder volume (V _SB, ml) and the fork length was: V _SB = 0.000213 FL³. The results from the in situ TS measurements indicated that the tilt angle was highly concentrated between −10° and 15°. The results from a calculation using the ex situ TS–FL equation with the fork length from biological sampling agreed strongly with the average in situ TS.     

Growth,diet, and reproduction of Eurasian perch <Emphasis Type="Italic">Perca fluviatilis</Emphasis> L. in Lake Varese,northwestern Italy

No data have previously been reported on Eurasian perch Perca fluviatilis L. in Lake Varese. In this study, the growth, diet, and reproductive biology of the Eurasian perch population were investigated with the aim of providing information that may serve as a basis for efficient resource management. A total of 240 specimens were caught during the monthly sampling campaign from November 2006 through October 2008. The length-to-weight relationships were W _t = 8.4 × 10⁻³ L _t^3.10 (males) and W _t = 4.1 × 10⁻³ L _t^3.36 (females). The parameters for the von Bertalanffy growth function for pooled sexes were L _∞  = 33.17 cm, k = 0.20 year⁻¹, and t ₀ = −1.34 year. Perch in Lake Varese spawn from April through May. Sexual maturity is reached when males are 2 years old, in females mostly when they are 3 years old. Relative fecundity (F _rel) and absolute fecundity (F _abs) were assessed for females. Fecundity values were similar to data reported for other European populations: females of age 2+ F _rel = 102,457 ± 12,275, age 3+ F _rel = 131,767 ± 5,891, and age 4+ F _rel = 131,252 ± 15,555. Perch diet spectrum was wide and somewhat characterized by season. Perch in Lake Varese feed on macroinvertebrates, mainly Chironomidae and Chaoborus, zooplankton, and juvenile rudd Scardinius erythrophthalmus.     

Effects of conventional versus organic production systems on amino acid profiles and heavy metal concentrations in the Chinese shrimp <Emphasis Type="Italic">Penaeus chinensis</Emphasis>

Amino acid composition and heavy metal concentration were examined in organic and conventional Chinese shrimp Penaeus chinensis production systems during the production cycle. The crude protein and total amino acid contents for organic shrimp samples were 0.932 and 0.945 mg/g d.w., while they were 0.849 and 0.826 mg/g d.w., respectively, for conventionally produced samples. The ratios of essential amino acids to nonessential amino acids in organic and conventional cultured shrimp were 0.58 and 0.63, respectively. Shrimp samples from the conventional system were significantly richer in Cu (2.28 vs. 1.53 mg kg⁻¹), Pb (0.13 vs. 0.08 mg kg⁻¹), Cd (0.09 vs. 0.04 mg kg⁻¹), and Zn (9.94 vs. 8.42 mg kg⁻¹) when compared with those obtained from the organic system. It is concluded that organic shrimp production systems can produce comparable growth and nutritional quality benefits.     

Protein and energy utilization and the requirements for maintenance in juvenile mulloway (<Emphasis Type="Italic">Argyrosomus japonicus</Emphasis>)

This study describes the digestible protein (DP) and digestible energy (DE) utilization in juvenile mulloway, and determined the requirements for maintenance. This was achieved by feeding triplicate groups of fish weighing 40 or 129 g held at two temperatures (20 or 26°C), on a commercial diet (21.4 g DP mJ DE⁻¹) at four different ration levels ranging from 0.25% of its initial body weight to apparent satiation over 8 weeks. Weight gain and protein and energy retention increased linearly with increasing feed intake. However, energy retention efficiency (ERE) and protein retention efficiency (PRE) responses were curvilinear with optimal values, depending on fish size, approaching or occurring at satiated feeding levels. Maximum predicted PRE was affected by body size, but not temperature; PRE values were 0.50 and 0.50 for small mulloway, and 0.41 and 0.43 for large mulloway, at 20 and 26°C respectively. ERE demonstrated a similar response, with values of 0.42 and 0.43 for small, and 0.32 and 0.34 for large mulloway at 20 and 26°C respectively. Utilization efficiencies for growth based on linear regression for DP (0.58) and DE (0.60) were independent of fish size and temperature. The partial utilization efficiencies of DE for protein (k _p) and lipid (k _l) deposition estimated using a factorial multiple regression approach were 0.49 and 0.75 respectively. Maintenance requirements estimated using linear regression were independent of temperature for DP (0.47 g DP kg^−0.7 day⁻¹) while maintenance requirements for DE increased with increasing temperature (44.2–49.6 kJ DE kg^−0.8 day⁻¹). Relative feed intake was greatest for small mulloway fed to satiation at 26°C and this corresponded to a greater increase in growth. Large mulloway fed to satiation ate significantly more at 26°C, but did not perform better than the corresponding satiated group held at 20°C. Mulloway should be fed to satiation to maximize growth potential if diets contain 21.4 g DP mJ DE⁻¹.     

Substrate specificity of brain acetylcholinesterase and its sensitivity to carbamate insecticides in <Emphasis Type="Italic">Carassius auratus</Emphasis>

The substrate specificity of brain acetylcholinesterase (AChE) in adult Carassius auratus fish and its sensitivity to carbamate insecticides were investigated in vitro. The results showed that the order of four substrates hydrolyzed by brain AChE in C. auratus was acetylthiocholine iodide > β-methylthiocholine iodide > propionylthiocholine iodide > butyrylthiocholine iodide, and the maximum velocity (V _max) of AChE hydrolyzing acetylthiocholine iodide (ATCh) was the highest among the four substrates, and the V _max values were 0.067 and 0.082 mmol min⁻¹ mg⁻¹ for male and female fish respectively. But their Michaelis–Menten constants (K _m) were the lowest, only 0.071 and 0.072 mmol/l respectively. Compared with other carbamate insecticides, the sensitivity of brain AChE to carbofuran was the highest and the IC₅₀ values were 1.04 × 10⁻⁶ mol/l for females and 1.17 × 10⁻⁶ mol/l for males. The inhibitory tendencies of eserine, methomyl, and aldicarb to brain AChE were very similar, and the percentage inhibition increased with time at the concentration of 1 × 10⁻⁶ mol/l. The order of inhibition potential of the three inhibitors from the highest to the lowest was eserine, aldicarb, and methomyl.     

Age and growth of two gizzard shads, <Emphasis Type="Italic">Nematalosa come</Emphasis> and <Emphasis Type="Italic">N. japonica</Emphasis>, in coastal waters around Okinawa Island,southwestern Japan

The age and growth of two Nematalosa species around Okinawa Island were examined using sectioned otoliths collected from September 2003 to April 2006. Monthly changes in the frequency of the appearance of a translucent band on the outer margin of the otoliths indicated that ring formation occurred once a year from January to July for Nematalosa come and from January to March for Nematalosa japonica. The von Bertalanffy growth equations for both species were as follows: N. come: L _t  = 365.5{1 − exp[−0.111 × (t + 0.288)]} for females and L _t  = 214.7{1 − exp[−0.700 × (t – 1.110)]} for males; N. japonica: L _t  = 205.1{1 − exp[−1.068 × (t − 1.180)]} for females and L _t  = 195.5 {1 − exp[−1.293 × (t − 1.269)]} for males. The maximum ages observed for N. come and N. japonica were 11 and 6 years old, respectively. The growth of these species was characterized by the slow growth of N. come over many years, resulting in a larger size than N. japonica.     

Comparative growth of Pacific oyster (Crassostrea gigas) postlarvae with microfeed and microalgal diets

Two trials were carried out in the laboratory in order to assess the effect of microparticulated feed (F) and live (Thalassiosira pseudonana, M) diets on the growth of recently set (396 ± 13 μm shell height) and 2 mm Crassostrea gigas postlarvae. Different proportions of M and F (100:0, 75:25, 50:50; 25:75, 0:100) were delivered in a single dose of 3 h d⁻¹ in trial 1. Dietary M:F proportions of 100:0, 50:50, and 0:100 were delivered as a single pulse of 8 h d⁻¹ (P1) or two pulses of 4 h⁻¹ (P2) in trial 2. Maximal daily M ration was 296 cells μl⁻¹ d⁻¹ (trial 1), 150 M cells μl⁻¹ d⁻¹ (trial 2), or their equivalent F dry weight. Shell height (SH), dry (DW), and organic weight (AFDW) were evaluated weekly. Oysters from trial 1 significantly increased their size after 28 days, and exhibited no significant dietary differences in terms of DW (1.21 ± 0.15 to 2.01 ± 0.28 mg) or AFDW (0.091 ± 0.022 to 0.166 ± 0.029 mg). Newly set postlarvae (trial 2) also exhibited significant growth after 25 days. No dietary differences were observed in trial 2, yet P2 oysters attained significantly higher shell heights (825–912 μm) than P1 oysters (730–766 μm) after 25 d. Pulse effects were marginally not significant in terms of AFDW and growth rate. Together, these findings showed that balanced microfeeds have a practical potential for the culture of early C. gigas postlarvae, when they are delivered in pulse-feeding schemes     

Potential predation rates by the sea stars Asterias rubens and Marthasterias glacialis, on juvenile scallops, Pecten maximus, ready for sea ranching

The potential for predation by the sea stars Asterias rubens and Marthasterias glacialis on seed-size (41 ± 3 mm shell height) juvenile scallops (Pecten maximus), ready for seeding in sea ranching areas, was investigated in a 30-day laboratory predation experiment. There was no significant difference (P > 0.05) in predation rate of large A. rubens (95–115 mm radium) and large M. glacialis (120–164 mm radius), which averaged 0.88 and 0.71 scallops individual⁻¹ day⁻¹, respectively. Maximum rates of predation were 2.44 scallops individual⁻¹ day⁻¹ for large A. rubens and 3.00 scallops individual⁻¹ day⁻¹ for large M. glacialis. Small M. glacialis (76–87 mm radius) had a significantly lower predation rate than large individuals of either species (average 0.13 scallops individual⁻¹ day⁻¹, P < 0.05). Small A. rubens (50–80 mm radius) only began to prey on scallops when average scallop size was reduced to 35 mm. Based on estimated density of sea stars at a Norwegian sea ranching site and average predation rates, a population of scallops seeded at 10 m⁻² would be reduced by between 0.5 and 11% in 1 month. Furthermore, using the highest observed predation rate, the degree of loss of scallops indicated that scallop culture via sea ranching would not be economically viable and thus methods for reducing scallop predation by sea stars are necessary.     

Trypsin from the viscera of Bogue (<Emphasis Type="Italic">Boops boops</Emphasis>): isolation and characterisation

Trypsin from the viscera of Bogue (Boops boops) was purified to homogeneity by precipitation with ammonium sulphate, Sephadex G-100 gel filtration and Mono Q-Sepharose anion exchange chromatography, with an 8.5-fold increase in specific activity and 36% recovery. The molecular weight of the purified enzyme was estimated to be 23 kDa by SDS–PAGE and size exclusion chromatography. The purified trypsin appeared as a single band on native-PAGE and zymography staining. The purified enzyme showed esterase-specific activity on N-α-benzoyl-l-arginine ethyl ester (BAEE) and amidase activity on N-α-benzoyl-dl-arginine-p-nitroanilide (BAPNA). The optimum pH and temperature for the enzyme activity, after 10 min incubation, were pH 9.0 and 55°C, respectively, using BAPNA as a substrate. The trypsin kinetic constants K _m and k _cat on BAPNA were 0.13 mM and 1.56 s⁻¹, respectively, while the catalytic efficiency k _cat /K _m was 12 s⁻¹ mM⁻¹. Biochemical characterisation of B. boops trypsin showed that this enzyme can be used as a possible biotechnological tool in the fish processing and food industries.