Effects of culture density and bottom area on growth and survival of the cuttlefish <Emphasis Type="Italic">Sepia</Emphasis><Emphasis Type="Italic">officinalis</Emphasis> (Linnaeus, 1758)

The effects of culture density and bottom areas on cuttlefish (Sepia officinalis) culture were studied. Cuttle fish were cultured under three experimental combinations of culture density and bottom area: (1) high density and small bottom surface area; (2) low density and large bottom area; (3) high density and large bottom area. Each experimental protocol was repeated in triplicate. Average weights at the end of the experiment were of 65.8 ± 5.8, 87.1 ± 5.6 and 78.7 ± 5.9 g for cuttlefish cultured under the conditions of protocol 1, 2, and 3, respectively; these differences were significant between each of the three groups. Total biomass increased faster (up to 7.5 kg per tank) under the high density/large bottom area conditions (protocol 3) due to the larger number of animals and low mortality in those tanks. Growth rates (%bw day⁻¹) were different between protocols, with growth rates of 2.1 ± 0.1, 3.0 ± 0.2, and 2.5 ± 0.1%bw day⁻¹ obtained for cuttlefish cultured according to protocol 1, 2, or 3, respectively. Average feeding rates were similar for the three groups—10.7 ± 0.8, 9.7 ± 2.0, and 9.6 ± 1.1%bw day⁻¹ for cuttlefish cultured according to protocols 1, 2, and 3, respectively, while food conversions (%) were different—21.5 ± 3.2, 32.4 ± 2.5, and 27.0 ± 1.1%bw day⁻¹, respectively. Total mortality was high in the high density/small bottom area tanks, 30%, while it was very low for the groups cultured under conditions of low and high density/large bottom area, 4%. Based on these results, we conclude that culture conditions that provide large bottom areas also provide good survival conditions and promote growth in comparison those with small bottom areas, even under conditions of lower culture densities.     

A preliminary study on the use of mannan oligosaccharides (MOS) in freshwater crayfish, <Emphasis Type="Italic">Astacus leptodactylus</Emphasis> Eschscholtz, 1823 juvenile diets

The effects of dietary mannan oligosaccharides (MOS) on the survival and growth of Astacus leptodactylus juveniles were evaluated. Experimental diets were prepared by using supplementation of 0 (Control), 1.5, 3.0, and 4.5 g MOS kg⁻¹ commercial trout larvae diet containing 43 g kg⁻¹ protein and 15.74 g kg⁻¹ lipid. A. leptodactylus juvenile with an average total length of 3.6 ± 0.46 cm (TL) and average weight of 1.25 ± 0.43 g was stocked in 0.2-m² aquariums at a rate of 50 crayfish/m² and reared for 60 days at 22.8°C. The experiment consisted of four treatments with three replicates each. Each aquarium contained ten crayfish. Crayfish juvenile fed with 0, 1.5, 3.0, and 4.5 MOS attained 6.15 ± 0.49, 5.94 ± 0.29, 7.34 ± 0.39, and 5.94 ± 0.27 cm final total length and 50, 56.67, 46.67, and 50% survival rates, respectively. Moreover, molting frequencies in 0, 1.5, 3.0, and 4.5 g kg⁻¹ MOS groups were detected as 44.44, 61.11, 83.33, and 38.88%, respectively. At the end of the experiment, a general enhanced growth performance (P < 0.05) and feed conversion ratio were observed in crayfish fed on the diet containing 3.0 g kg⁻¹. Crayfish juveniles fed with 3.0 g kg⁻¹ MOS had the highest final length (7.34 ± 0.39 cm), although their survival was the lowest (46.67%) compared to other treatments but not significant. Specific growth rates were significantly different (P < 0.05) among the treatment groups at the end of the 60-day experiment. There were no significant differences (P > 0.05) in percentage moisture, protein, and ash (wet-weight basis) in the tail meat of A. leptodactylus juvenile among treatments (diet) that averaged 82.3, 16.1, and 1.2%, respectively. However, lipid value was significantly different among the diets P < 0.05), with values between 0.13 and 0.32. From the current results, it could be advised to use MOS at the rate of 3.0 g kg⁻¹ in A. leptodactylus juvenile diets.     

Stocking density for the intensive rearing of juvenile crayfish, <Emphasis Type="Italic">Pacifastacus leniusculus</Emphasis> (Astacidae), using <Emphasis Type="Italic">Artemia</Emphasis> nauplii to supplement a dry diet from the onset of exogenous feeding

Recent advances in intensive rearing of astacid juvenile crayfish have greatly improved the results. This challenges the current application possibilities of the studies performed previously, and new research on density is required. A 100-day experiment was carried out under controlled conditions to evaluate density effects on survival and growth rates of juvenile crayfish in optimal conditions of feeding. Juvenile stage 2 Pacifastacus leniusculus were stocked in fibreglass tanks (1 m², 200 l water) at 20 ± 1°C and fed a dry diet for salmonids supplemented with restricted amounts of Artemia nauplii. Stocking densities were 100, 300, 600 and 1,000 crayfish m⁻². Mean survival rate was reduced significantly with increased stocking density, ranging from 86.33% (100 m⁻²) to 39.13% (1,000 m⁻²). All checks showed that at the lowest initial density (100 m⁻²) animals grew significantly faster those at higher densities, recording a final carapace length of 15.28 mm and weight of 1.08 g. Among the treatments of 300, 600 and 1,000 m⁻² no differences were found either in carapace length or in weight throughout the experimental period, with a final mean growth of 14 mm carapace length and 0.72 g weight. The final proportion of animals with chelae autotomy rose significantly with increasing stocking density, ranging from 14.44% (100 m⁻²) to 41.45% (1,000 m⁻²). This study shows that diet is a decisive factor for stocking successfully high densities under controlled conditions and provides useful information to set adequate densities in accordance with the production objectives.     

Effects of two dietary protein levels on energy balance and digestive capacity of <Emphasis Type="Italic">Octopus maya</Emphasis>

Octopus maya is a carnivorous species and protein is the main energy source. During the present study, two different dietary protein levels (40 and 60% CP) were offered to octopuses as specifically designed artificial diets, to determine protein needs and the effects on metabolism. Frozen crab (Callinectes spp.) was used as control. Results obtained demonstrated that crab remains as one of the best diets for O. maya. The artificial diet with 60% CP produced a low but positive growth rate, and at times, a physiological response similar to that observed in octopuses fed crabs. The present results show the capacity of O. maya juveniles to adjust their digestive enzymes to different types of food and protein level, and this appears to be well correlated with octopus growth. General proteases and trypsin from the pancreas were well correlated with growth rates. A low activity was observed in octopuses fed 40% PC diet (negative growth rate), while a high activity was present in octopuses fed 60% CP diet and crabs (low and high growth rate, respectively). In contrast, these same enzymes were inducted in the salivary glands of octopuses fed with the diet that promoted weight loss (40% CP diet), while a reduced activity was observed in octopuses fed crabs. Energy budget indicates that the animals ingested more than 1,000 kJ week⁻¹ kg⁻¹; with such energy, octopuses should satisfy their physiological demands such as was observed when animals were fed crab (I = 1,300 kJ week⁻¹ kg⁻¹; P = 834 kJ week⁻¹ kg⁻¹). However, a very low digested energy was observed in octopuses the fed artificial diets, indicating that these could have a factor that limits digestibility.     

Influence of natural diet on growth and biochemical composition of <Emphasis Type="Italic">Octopus vulgaris</Emphasis> Cuvier, 1797

The influence of diet on growth and biochemical composition of cultured Octopus vulgaris was investigated. Octopuses were reared in a open recirculation seawater system and during the experiment were kept apart in cages. Experiment lasted 30 days. Three experimental dietary groups were established: group I (mixed diet: Carcinus mediterraneus, Boops boops and Mytilus galloprovincialis), group II (monodiet on B. boops) and group III (monodiet on M. galloprovincialis). The best results were obtained with the monodiet based on bogue that gave a weight gain of 390.65 ± 37.54, an absolute growth rate of 13.02 g day⁻¹ and feed efficiency of 44.79 ± 0.96%. Regarding to biochemical composition, the mussel-fed and bogue-fed groups showed the highest protein content. Animals fed on bogue also showed the highest carbohydrate content, while the lowest value was observed in the mussels-fed group. The lipid content was found to be very low in all dietary groups. Saturated fatty acids were the most abundant fatty acid class, ranging from 58.22% in wild octopuses to 46.83% in bogues-fed octopuses. Polyunsaturated fatty acids (PUFAs) were the second most abundant fatty acids class in all dietary treatments (25.65–37.00% as a per cent of total FAs) except in octopuses fed with mussels. MUFAs ranged from 11.44 to 26.93% of total FAs. Although it is empirically known that a varied diet better covers the nutritional requirements than a monodiet, the results of this study reveal that a monodiet of B. boops may be used.     

Optimum ration level for better growth,conversion efficiencies and body composition of fingerling <Emphasis Type="Italic">Heteropneustes fossilis</Emphasis> (Bloch)

The effects of ration levels on growth, conversion efficiencies and body composition of fingerling Heteropneustes fossilis (6.8 ± 0.04 cm, 5.0 ± 0.02 g) were studied by feeding isonitrogenous (40% crude protein) and isocaloric (19.06 MJ kg⁻¹ gross energy) diets representing 1, 3, 5, 7 and 9% of the body weight (BW) day⁻¹ to triplicate groups of fish . Growth performance of the fish fed at the various ration levels was evaluated on the basis of live weight gain percentage (LWG%), feed conversion ratio (FCR), specific growth rate percentage (SGR%), protein retention efficiency (PRE%) and energy retention efficiency (ERE%) data. Maximum LWG% and SGR were obtained at a feeding rate of 7% BW day⁻¹, whereas best FCR (1.6), PRE% and ERE% were recorded at a feeding rate of 5% BW day⁻¹. Maximum body protein was also obtained for the group receiving the diet representing 5% of their body weight. However, a linear increase in fat content was noted with the increase in ration levels up to 7% BW day⁻¹. The SGR, FCR, PRE and ERE data were also analyzed using second-degree polynomial regression analysis to obtain more precise information on ration level, with the results showing that the optimal ration for these parameters was 6.8, 6.1, 5.9 and 6.2% BW day⁻¹, respectively. Based on the above second-degree polynomial regression analysis, the optimum ration level for better growth, conversion efficiencies and body composition of fingerling H. fossilis was found to be in the range of 5.9–6.8% of the BW day⁻¹, corresponding to 2.36–2.72 g protein and 88.20–101.66 MJ digestible energy kg⁻¹ diet day⁻¹.     

The effect of stocking density on yield,growth, and survival of Asian river catfish (<Emphasis Type="Italic">Pangasius</Emphasis><Emphasis Type="Italic">bocourti</Emphasis> Sauvage, 1880) cultured in cages

Asian river catfish (Pangasius bocourti Sauvage, 1880) were cultured at five different stocking densities in cages (submerged volume 1 m³) suspended in a dugout pond from August to November 2009. Pangasius bocourti fingerlings (mean weight 27.09 ± 0.54 g) were stocked at densities of 12, 25, 50, 100, and 200 fish m⁻³. At the end of 3 months, the harvest weights (gross yields) were, respectively, 2.05 ± 0.30, 5.20 ± 0.31, 10.60 ± 0.42, 19.98 ± 0.78, and 42.37 ± 0.41 kg m⁻³. The mean fish weights among the stocking densities of 25, 50, 100, and 200 fish m⁻³ were not significantly different, but were significantly higher than that of the 12 fish m⁻³ density. The specific growth rates among high stocking densities of 50, 100, and 200 fish m⁻³ were not significantly different; however, they were significantly higher than those of the low stocking densities of 12 and 25 fish m⁻³. Asian river catfish performed poorly at the lowest density. The results indicate an initial lower stocking threshold for Asian river catfish of above 5.20 kg m⁻³. The Asian river catfish cultured in small cages placed in a pond reached the desirable market size (>200 g) within a 90-day grow-out period. The results show that the maximum yield for Asian river catfish during a 3-month production cycle was not reached.     

Growth,absorption and assimilation efficiency by mature cuttlefish (<Emphasis Type="Italic">Sepia officinalis</Emphasis>) fed with alternative and artificial diets

The effects of feeding three natural frozen diets, grass shrimp (Palaemonetes sp.), crayfish (Procambarus clarkii) and fish (Sardina pilchardus) and two semi-humid artificial diets (based on fish powder) to mature cuttlefish, Sepia officinalis, were analysed. Growth and feeding rates (GR and FR, % BW day⁻¹), food conversions (FC, %), and total protein and lipid composition of the diets were determined. Digestive gland to body weight ratio and absorption efficiency were calculated for each diet. Cuttlefish fed shrimp and crayfish grew larger (1.5 and 1.1% BW day⁻¹, respectively) compared to the other diets. Shrimp promoted the highest FC, followed by crayfish, and sardine. The highest FR was obtained for cuttlefish fed crayfish (8.4% BW day⁻¹). Although both artificial diets were accepted, none produced growth. A positive correlation (r = 0.96) between cuttlefish ingestion rate and digestive gland weight was obtained. Some cannibalism occurred among cuttlefish fed the artificial diets during the last week of the experiment. According to the results obtained, P. clarkii could be used as an alternative prey to shrimp for rearing adult mature (>50 g) S. officinalis.     

Effects of three culture densities on growth and survival of <Emphasis Type="Italic">Octopus vulgaris</Emphasis> (Cuvier, 1797)

The purpoase of this research was to test the effects of three culture densities on the growth and survival of Octopus vulgaris. A total of 141 sub-adult octopuses (1,175.4 ± 194.9 g) were randomly distributed in nine tanks of 2,000 l each (3.6 m × 1.1 m, and 50 cm water depth). Three tanks were stocked with a low initial density of 4 kg/m³, while three other tanks were stocked with an initial density of 8 kg/m³, and the remaining three tanks were stocked at an initial density of 15 kg/m³. Octopuses were all fed frozen squid (Loligo gahi) at 5% body weight per day (%BW/day). The experiment lasted for 70 days. Water temperature varied between 20 ± 2°C, and salinity varied between 36 ± 1 ppt. During the entire experiment, dissolved oxygen was always >75%, and ammonia was always lower than 0.1 mg/l. No differences in growth or growth rates (between 0.9 and 1.1%BW/day for the three densities) were found. Nevertheless, mortality was significantly lower for the low density compared to the other two densities tested. Maximum densities in the culture tanks (>25 kg/m³) were attained in the higher culture densities after 56 days of the experiment.     

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.     

Effect of a probiotic bacterium <Emphasis Type="Italic">Bacillus circulans</Emphasis> PB7 in the formulated diets: on growth,nutritional quality and immunity of <Emphasis Type="Italic">Catla catla</Emphasis> (Ham.)

Bacillus circulans PB7, isolated from the intestine of Catla catla, was evaluated for use as a probiotic supplement in the feeds for the fingerlings of Catla catla. The effect of supplement on the growth performance, feed utilization efficiency, and immune response was evaluated. Catla fingerlings (ave. wt. 6.48 ± 0.43 g) were fed diets supplemented with 2 × 10⁴ (feed C1), 2 × 10⁵ (feed C2), and 2 × 10⁶ (feed C3) B. circulans PB 7 cells per 100 g feed for 60 days at 5% of the body weight per day in two equal instalments in triplicate treatments. The control feed (CC) was not supplemented with the B. circulans. All the feeds were isocaloric and isonitrogenous. Fish fed with feed C2 displayed better growth, significantly (P ≤ 0.05) highest RNA/DNA ratio, a lower feed conversion ratio (FCR), and a higher protein efficiency ratio (PER) than the other experimental diets. Highest carcass protein and lipid was also observed in the fish fed C2 feed compared to the others. Significantly (P ≤ 0.05), highest protease was recorded in fish fed feed C2 (47.9 ± 0.016) and lowest in fish fed feed C3 (32.10 ± 0.009), where α-amylase activity did not differ significantly (P ≤ 0.05) beyond the lowest inclusion level. ALP, ACP, GOT, and GPT in the liver of Catla catla were the highest (P ≤ 0.05) in fish fed C2 feed. The highest TSP, albumin, and globulin was observed in fish treated with C2 feed after 60 days feeding trial, but the lowest glucose level was observed in the same treatment. After the feeding trial, the non-specific immunity levels and disease resistance of fish were also studied. Phagocytic ratio, phagocytic index, and leucocrit value were the highest in fish fed feed C2. After the feeding trial, the fish were challenged for 10 days by bath exposure to Aeromonas hydrophila (AH1) (10⁵c.f.u. ml⁻¹ for 1 h, and, after 7 days, 10⁷c.f.u. ml⁻¹ for 1 h). Highest survival percentage was observed in fish fed with feed C2 compared with only 6.66% in the controls, which indicated the effectiveness of B. circulans PB 7 in reducing disease caused by A. hydrophila.     

Effects of dietary calcium and phosphorus supplementation on the growth performance of juvenile spotted babylon <Emphasis Type="Italic">Babylonia areolata</Emphasis> culture in a recirculating culture system

A feeding experiment was conducted to determine the effects of dietary calcium and phosphorus, and the interaction between calcium and phosphorus, on the growth of juvenile spotted babylon, Babylonia areolata, cultured in a recirculating culture system. Nine isonitrogenous experimental diets supplemented with three levels of calcium (1, 4, and 7%) for each of three levels of phosphorus (1, 3, and 5%) were prepared using fish meal, squid meal, and shrimp meal as the main protein sources. Juveniles with an initial average body weight of 0.59 ± 0.09 g were fed to satiation once daily with one of the nine diets for 180 days. Absolute and specific growth rates were calculated for both shell length and whole wet body weight. Results showed that dietary calcium and phosphorus supplementation significantly affected the growth of juvenile spotted babylon (P < 0.05), but not survival and feed-conversion ratio. The specific growth rate in shell length (SGRL) ranged from 0.32 to 0.39% day⁻¹. No significant difference among phosphorus levels and no significant interaction between calcium and phosphorus in SGRL of the spotted babylon (P > 0.05) was found, but significant differences were observed among calcium levels, irrespective of phosphorus levels (P < 0.05). For 1 and 7% supplemental calcium, the spotted babylon had significantly higher SGRL than those fed diets supplemented with 4% calcium. However, the specific growth rate in body weight (SGRW) ranged from 0.91 to 1.19% day⁻¹ with no significant difference among calcium and phosphorus levels and no significant interaction between calcium and phosphorus (P > 0.05). Survival and feed-conversion ratio were not significantly affected by dietary calcium and phosphorus levels with ranges from 91.00 to 95.00% and 2.43 to 2.76, respectively. At the end of the experiment, shell abnormality of B. areolata was found for all feeding trials.     

Effect of dietary chromium picolinate on growth performance and blood parameters in grass carp fingerling, <Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis>

An experiment was conducted to investigate the effect of dietary chromium picolinate supplement on growth and haematology parameters of grass carp, Ctenopharyngodon idellus. Six diets with increasing dietary chromium picolinate levels 0, 0.2, 0.4, 0.8, 1.6 and 3.2 mg kg⁻¹ were fed to triplicate groups of 20 fish (initial weight of 12.78 ± 1.16 g, mean ± SD) in a flow water system for 10 weeks. Fish fed the diet supplemented with 0.8 mg Cr kg⁻¹ had significantly improved weight gain (WG), feed efficiency ratio (FER), protein efficiency ratio (PER) and protein retention (PR). Fish fed high-chromium diets exhibited lower whole-body crude lipid contents than fish fed low-chromium diets. Liver glycogen concentrations for fish fed the diet with 0.2 mg Cr kg⁻¹ was the highest (77.67 mg g⁻¹). Fish fed the diet supplemented with 1.6 and 3.2 mg Cr kg⁻¹ had significantly lower liver glycogen concentrations than other groups (P < 0.05). The highest serum insulin concentrations were observed in fish fed the diet supplemented with 0.8 mg Cr kg⁻¹, but serum insulin concentrations decreased (P < 0.05) when dietary supplementation of chromium was higher than 0.8 mg Cr kg⁻¹. Cholesterol concentrations decreased in direct proportion to dietary chromium level and achieved the lowest level when the fish were fed the 0.8 mg Cr kg⁻¹ diet, but increased when the fish were fed the diet with more than 0.8 mg Cr kg⁻¹ (P < 0.05). Fish fed the diet supplemented with 0.8 mg Cr kg⁻¹ had higher triglyceride and high-density lipoprotein cholesterol (HDL-C) concentrations compared to other treatments. The results of the present study suggested that chromium picolinate could modify serum carbohydrate and lipid metabolism profile, and that the optimal dietary chromium level was 0.8 mg kg⁻¹ for grass carp according to growth.     

Probiotic bacteria <Emphasis Type="Italic">Lactobacillus rhamnosus</Emphasis> influences the blood profile in rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis> (Walbaum)

This paper reports the effect of feeding probiotic diets on blood profiles in rainbow trout. Two experiments were performed: in the first, fish of average weight 75 g were offered either a commercial feed or the same incorporated with 10⁹ CFU g⁻¹ of lactic acid bacteria Lactobacillus rhamnosus for 30 days; in the second study performed for a similar duration, fish of average weight 126 g were offered formulated diets that either contained the same bacteria in heat-killed or freeze-dried form (nearly 10¹¹ CFU g⁻¹), or the basal diet without the bacteria. Blood samples were collected at different times after commencement of probiotic feeding to determine the total cholesterol, triglyceride contents, the plasma alkaline phosphatase activity, plasma protein and hematocrit value. The plasma cholesterol significantly increased upon probiotic feeding in the first experiment. A significant elevation (P < 0.05) of plasma cholesterol and triglyceride and alkaline phosphatase activity level was found in the freeze-dried probiotic fed groups at 20 and 30 days postfeeding. This was concomitant with the increased plasma protein and hematocrit values in FD group at 20 and 30 days. Likewise, the heat-killed probiotic fed group registered significantly high values of triglycerides, alkaline phosphatase activity, and plasma protein compared to the control diet fed groups after 20 days of feeding. Thus, alterations in the blood profiles could serve as supplementary information when examining the benefits of probiotics for fish.     

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.     

Growth and survival of grouper <Emphasis Type="Italic">Epinephelus</Emphasis><Emphasis Type="Italic">coioides</Emphasis> (Hamilton) larvae fed free-living nematode <Emphasis Type="Italic">Panagrellus redivivus</Emphasis> at first feeding

The free-living nematode, Panagrellus redivivus, was tested as live food for grouper Epinephelus coioides larvae during the first feeding stage. A series of experiments were conducted to determine the acceptability of the free-living nematodes in grouper larvae at first feeding, the optimum nematode density and the response of the larvae to nutritionally enriched nematode. All experiments were conducted in 200-L conical tanks filled with 150-L filtered seawater and stocked at 15 larvae L⁻¹. Duration of feeding experiments was up to day 21 (experiment 1) and 14 days (experiment 2 and 3). Brachionus plicatilis and Artemia (experiment 1) and Brachionus plicatilis alone (experiment 2 & 3) was used as the control treatment. Observations indicated that the grouper larvae readily fed on free-living nematodes as early as 3 days posthatching, the start of exogenous feeding. Optimum feeding density for the larvae was 75 nematodes ml⁻¹. The enrichment of cod liver oil or sunflower oil influenced the total lipids and n-3 highly unsaturated fatty acids of P. redivivus, which in turn influenced those of the grouper larvae, however, growth and survival of the larvae were not affected (P > 0.05). The results from this investigation showed that the nematode, P. redivivus, can be used as first live food for grouper larvae from the onset of exogenous feeding until they could feed on Artemia nauplii.     

Swimming angle and target strength of larval Japanese anchovy (<Emphasis Type="Italic">Engraulis japonicus</Emphasis>)

The swimming angle of larval Japanese anchovy (Engraulis japonicus) was measured in a tank, and target strength (TS) was calculated using a theoretical scattering model. The mean swimming angle was 12.8° (SD ±22.1). Increased speeds of flow led to increased mean swimming angles. The mean swimming angle at flow of 5 cm s⁻¹ was higher than at other speeds. TS values were estimated using a distorted-wave Born approximation model for two cases. Average values were 1–3 cm s⁻¹ (11.5° ± 22.1) and 5 cm s⁻¹ (16.6° ± 21.7) for cases 1 and 2, respectively. For case 1, TS ranged from −92.0 to −74.7 dB with a mean of −79.4 dB at 120 kHz. For case 2, TS ranged from −92.2 to −75.2 dB with a mean of −79.9 dB. The mean TS in case 2 was lower than that in case 1, with the maximum difference being 1.0 dB at 120 kHz (standard length 22.0 mm). However, there were no significant differences between the regression lines of cases 1 and 2. Thus, changes in flow speed altered the swimming angle of larval Japanese anchovy, but had little influence on TS.     

Dietary polysaccharide from <Emphasis Type="Italic">Angelica sinensis</Emphasis> enhanced cellular defence responses and disease resistance of grouper <Emphasis Type="Italic">Epinephelus malabaricus</Emphasis>

The purpose of this study was to determine the effect of Angelica sinensis polysaccharide (ASP) supplemented in diet on the innate cellular immune response and disease resistance in grouper, Epinephelus malabaricus. Fish were fed diets containing different doses of ASP (0, 500 and 3,000 mg kg⁻¹ diet) for 12 weeks. After 12 week feeding, the respiratory burst activity, phagocytic activity, and leukocytes proliferation in head kidney were assayed. The functional immunity in terms of cumulative mortality was also assessed by a challenge with live Edwardsiella tarda. Results showed that the respiratory burst activities in ASP-supplemented groups were increased significantly. The respiratory burst index was the highest in fish-fed 3000 mg kg⁻¹ diet and the lowest in control. The phagocytic activities in ASP-supplemented groups were significantly higher than that of control. No significant difference in phagocytic activity was observed between ASP-supplemented groups. ASP stimulated the head kidney leukocytes proliferation significantly, despite the absence of lipopolysaccharide (LPS) or not. The cumulative mortalities of fish fed with 3000 mg ASP kg⁻¹ diet were significantly lower than those fed with 500 mg ASP kg⁻¹ diet and control diet after 96 h of challenge. In conclusion, dietary ASP enhanced some cellular immune parameters and disease resistance against E. tarda in grouper.     

Effects of dietary lipid levels on growth performance,feed utilization and fatty acid composition of juvenile Japanese seabass (<Emphasis Type="Italic">Lateolabrax japonicus</Emphasis>) reared in seawater

Triplicate groups of 40 Japanese seabass Lateolabrax japonicus (initial weight, 11.3 ± 0.4 g) reared in seawater (salinity, 30.0–33.0 g L⁻¹) were fed with five isonitrogenous (41.3 ± 0.2% crude protein) and isoenergetic (18.5 ± 0.3 MJ kg⁻¹) experimental diets formulated with increasing lipid levels (4.3, 8.4, 12.2, 15.8 and 20.1% lipid) for 10 weeks. Survival throughout the feeding experiment ranged from 87.5 to 100%, but the survival of fingerlings fed with the 4.3% lipid diet was significantly lower than the rest of the diets. At the end of the feeding experiment, fish fed with 12.2% lipid diet showed optimal growth performance (P < 0.05). Lipid contents of whole body, liver and muscle increased in parallel with the increase in dietary lipid levels. Viscerosomatic index (VSI), hepatosomatic index (HSI) and muscle lipid content were higher in 20.1% lipid group than those in the rest of the lipid level groups indicating that viscera and muscle tissues played important contributions to body lipid deposition. High proportions of 18:1n-9, eicosapentaenoic acid (20:5n-3; EPA) and docosahexaenoic acid (22:6n-3; DHA), and low concentrations of n-6 polyunsaturated fatty acids (PUFAs) occurring in liver and muscle, to some extent, reflected fatty acid (FAs) composition in the experimental diets.     

The effects of continuous photoperiod (24L:0D) on growth of juvenile barramundi (<Emphasis Type="Italic">Lates calcarifer</Emphasis>)

The efficacy of photoperiod manipulation to influence growth and developmental processes is well documented in a range of temperate aquaculture species. However, the application of such techniques with tropical species requires further investigation. This preliminary 20-day study investigated the influence of continuous photoperiod on growth of barramundi (Lates calcarifer). In addition, diel plasma melatonin profiles provided a physiological measure of how the endocrine system of barramundi responded to continuous photoperiod. Juvenile barramundi (1.33 ± 0.02 g) were held in recirculation systems under 12-h light: 12-h dark (12L:12D) or 24-h light (24L:0D) with a light intensity of 1,000 lux throughout the water column. Fish from both treatments grew to more than 14 times their original weight, with final weight (24L:0D = 21.59 ± 0.85 g; 12L:12D = 19.12 ± 0.55 g), total length (24L:0D = 12.67 ± 0.14 cm; 12L:12D = 11.96 ± 0.13 cm) and specific growth rate (24L:0D = 9.60 ± 0.05% bw day⁻¹; 12L:12D = 9.14 ± 0.06% bw day⁻¹) being significantly higher for fish grown on 24L:0D compared with 12L:12D. There were no significant differences in feed intake (24L:0D = 226.46 ± 6.27 g; 12L:12D = 219.02 ± 5.73 g) or feed conversion ratio (24L:0D = 0.71 ± 0.06; 12L:12D = 0.80 ± 0.07) between light treatments. Barramundi held under 12L:12D exhibited diel melatonin secretion, which peaked mid-dark phase (171.83 ± 4.81 pg ml⁻¹) followed by a gradual decrease in base levels at the onset of illumination (68.61 ± 8.77 pg ml⁻¹). When juvenile barramundi were subjected to 24L:0D, the amplitude of peak melatonin secretion was significantly suppressed during the subjective mid-dark phase (129.71 ± 2.36 pg ml⁻¹). This preliminary study confirmed that barramundi respond to photoperiod manipulation in a similar manner to many temperate fish species, thus demonstrating the future potential use of artificial lighting to improve growth in this species commercially.