Protein requirement of silver barb, Puntius gonionotus fingerlings

Five iso‐energetic (15.05 MJ kg^?1) semi‐purified diets with graded levels of crude protein, i.e. 200 (D‐1), 250 (D‐2), 300 (D‐3), 350 (D‐4) and 400 (D‐5) g kg^?1 diet were fed to Puntius gonionotus fingerlings (average weight 0.88 ± 0.03 g) in triplicate groups (15 healthy fish per replicate) for a period of 90 days to determine the optimum protein requirement of the fish. Fifteen flow‐through cement tanks of 100‐L capacity with a flow rate of 0.5 L min^?1 were used for rearing the fish. Specific growth rate (SGR), food conversion (food gain) ratio (FCR), nutrient digestibility and retention, digestive enzyme activity, RNA : DNA ratio and tissue composition were used as response parameters with respect to dietary protein levels and feed intake. The mean weight gains of fish after 90 days were 10.84 ± 0.27, 11.07 ± 0.12, 14.09 ± 0.20, 11.27 ± 0.12 and 10.91 ± 0.25 g for D‐1, D‐2, D‐3, D‐4 and D‐5, respectively. Maximum SGR (3.13 ± 0.02% per day), RNA : DNA ratio (10.09 ± 0.09), tissue protein content (160 ± 0.1 g kg^?1 wet weight), protease activity (25.27 ± 0.47 μg of leucine liberated mg tissue per protein h^?1 at 37 °C) and minimum FCR (1.60 ± 0.02) was found in D‐3 group fed with 300 g kg^?1 protein level. All these parameters were negatively affected with the further increase in protein level in the diet. Digestibility of protein, lipid and energy was not affected because of variation in dietary protein levels and nitrogen intake of fish. Maximum energy retention (27.68 ± 0.12%) was recorded at 300 g kg^?1 dietary crude protein fed group. However, using broken line regression analysis, the maximum growth was found to be at 317.7 g kg^?1 dietary protein. Hence, it may be concluded that the protein requirement of P. gonionotus fingerling is 317.7 g kg^?1 diet with a resultant P/E ratio of 21.1 g protein MJ^?1.     

Interacting effects of dietary lipid level and temperature on growth, body composition and fatty acid profile of rohu, Labeo rohita (Hamilton)

Three isonitrogenous (320 g kg^?1 crude protein, casein and gelatine) semi‐purified diets with 80 (L8), 130 (L13) and 180 (L18) g kg^?1 lipid (sunflower oil at increasing levels and cod liver oil fixed at 50 g kg^?1) at three digestible energy levels (12 096, 13 986 and 15 876 kJ kg^?1 dry weight) and were tested, in triplicate, on rohu fingerlings (3.2 ± 0.08 g) at two different temperatures (21 and 32 °C). Fish were fed to apparent satiation, twice daily, at 09.00 and 15.00 h, 7 days a week for 56 days. Maximum growth was obtained at a lipid level of 80 g kg^?1 (L8) at 21 °C (439.37%) and 130 g kg^?1 (L13) at 32 °C (481.8%). In general growth rate was higher at 32 °C than at 21 °C at all lipid levels. Tissue monounsaturated fatty acid (MUFA) contents decreased with increasing lipid level at 32 °C, but the reverse occurred at 21 °C. At 21 °C, Polyunsaturated fatty acid (PUFA) level increased significantly (P > 0.05) over initial values, but was affected insignificantly by dietary lipid level. At 32 °C, fish fed diet L13 had more n‐3 fatty acid (FA) in liver and muscle than the other two dietary groups while at 21 °C, both liver and muscle FA profiles exhibited significant change (P > 0.05) in n‐3 and n‐6 FA content which corresponded to variation in percent addition of dietary lipid. However, n‐3/n‐6 ratio was higher for fish fed diet L13 at 32 °C and diet L8 at 21 °C and may be correlated with fish growth.     

Partial replacement of fish oil by flaxseed oil in Atlantic halibut (Hippoglossus hippoglossus L.) diets: effects on growth,nutritional and sensory quality

Three isonitrogenous (520 g protein kg^?1 DM) and isoenergetic (25 MJ kg^?1 DM) diets containing increasing levels of flaxseed oil (FxO; 0%, 40% and 70% of total added oil) at the expense of fish oil (FO) were tested for 33 weeks in groups of 61 individually PIT‐tagged halibut (initial weight, 849 ± 99 g). Effects on fish growth performance, fillet nutritional and sensory quality were determined. Specific growth rate (0.2% day^?1), feed conversion ratio (1.2–1.3) and nitrogen and energy retention were not affected by dietary treatments. Dietary fatty acid composition was reflected in fatty acid profiles of halibut muscle, liver and heart. Muscle of fish fed FxO diets contained higher 18:2n‐6 and 18:3n‐3 concentrations whereas 20:5n‐3 and 22:6n‐3 levels were significantly reduced. However, increasing FO replacement induced preferential retention of 22:6n‐3 especially in heart, and a trend for 20:5n‐3 conservation in heart and muscle was observed. FO replacement did not affect colour, texture and the characteristic fish odour and flavour of cooked fillets. By selectively retaining long‐chain polyunsaturated fatty acids halibut can adapt to a lower dietary supply without adverse effects on growth, feed conversion ratio, survival, and fillet nutritional and sensory quality.     

Growth of young cuttlefish, Sepia officinalis (Linnaeus 1758) at the upper end of the biological distribution temperature range

Individual growth rates, feeding rates (%BWd^?1) and food conversions for cuttlefish (S. officinalis) hatchlings and juveniles were determined during this study. A flow‐through system was used. Water temperature reached 30 °C during the hottest part of the day, gradually decreasing to 25 °C during the night; salinity varied between 37 ± 3 ppt and lights were kept on for 14 h day^?1. Hatchlings were placed in separate compartments with a water volume of 1.2 L. Juvenile cuttlefish (from 0.5 to 25 g) were placed in bigger baskets, with a water volume of 5.2 L. Water flow was 120 L h^?1. The biggest cuttlefish used in these experiments (> 25 g) were gathered in groups of five and placed in circular tanks (water volume of 250–300 L). Thus, results obtained in this case are means and not individual data. During the first 10, 20, 30 and 40 days, mean growth rates (of all individuals sampled by age group) decreased consistently (11.8 ± 4.1, 9.8 ± 1.8, 8.1 ± 2.2 and 7.3 ± 0.7%BW^?1 respectively); in similar fashion, mean feeding rates decreased with age group (33.7 ± 13.5, 22.0 ± 7.9, 17.3 ± 3.9 and 16.7%BWd^?1 respectively). Mean food conversions varied between 3.6 and 2.5 between the age groups. When grouping results by weight class, similar patterns occur, as growth and feeding rates decrease consistently as cuttlefish grow bigger. Highest mean growth and feeding rates are obtained by hatchlings (< 0.1 g) with 12.4 ± 4.5 and 35.3 ± 15.1%BWd^?1, respectively, while the lowest growth and feeding rates were recorded for the largest animals, between 15 and 25 g (3.4 ± 1.1 and 10.8 ± 4.1%BWd^?1 respectively). For these weight classes, mean food conversions varied between 2.7 ± 0.9 and 3.8 ± 2.8.     

Protein lipid ratio for the growth and survival of juvenile crayfish Procambarus acanthophorus (Hobbs 1972) reared under controlled conditions

The influence of dietary protein/lipid levels on growth and survival in juvenile crayfish (Procambarus acanthophorus) was evaluated during a 12‐week nutritional trial. Twenty experimental diets were formulated containing different protein (200, 250, 300, 350 and 400 g kg^?1) and lipid (60, 80, 100 and 120 g kg^?1) levels. A bifactorial model (5 × 4) was used with three replicates and 15 crayfish (0.014±0.006 g) per replicate, in a recirculation system with 60–15 L rectangular tanks and a biological filter, the culture conditions were a photoperiod of 12:12 h L:D and temperature 24.7±1.0 °C. Total survival ranged from 66.6% to 86.6%, with no differences attributed to treatments. Dietary protein levels had a significant effect on growth gain and nutritional efficiency; in contrast, lipid levels had no effect on juvenile performance. The 250/120 (protein/lipid) treatment promoted the most efficient growth (final weight: 3.09 g, weight gain: 21 995%, specific growth rate: 3.35% day^?1); the less efficient growth was observed in juveniles fed with 400 g kg^?1 protein diets. The quadratic regression model indicated 279 g kg^?1 as the optimal protein level for crayfish growth, while levels between 211 and 232 g kg^?1 could minimize the feeding costs without affecting growth.     

Simultaneous detection of pathogens causing francisellosis,furunculosis and vibriosis in Atlantic cod by multiplex polymerase chain reaction

An assay for the simultaneous detection of three major bacterial pathogens of Atlantic cod with multiplex polymerase chain reaction (mPCR) was developed. Primers targeting the flanking regions of genes groEL, gyrB and amiB were standardized to diagnose francisellosis, furunculosis and vibriosis respectively. The detection limit of Francisella piscicida, Aeromonas salmonicida and Vibrio anguillarum in the mPCR assay ranged from 10 ng to 100 pg of bacterial DNA mL^?1, 10 μg to 50 ng of bacterial DNA mL^?1 and 1 μg to 100 ng of bacterial DNA mL^?1 respectively. These primers were found to be specific in the determination of the respective genes in the individual pathogen without cross‐reaction, thus making the assay an efficient tool to detect the presence of these pathogens simultaneously or individually.     

Protein-sparing effect of carbohydrate in silver barb, Puntius gonionotus fry

A 30‐day study was undertaken to examine the protein‐sparing effect of carbohydrate in diets for silver barb, Puntius gonionotus fry. Six semi‐purified experimental diets were formulated with two levels of protein (200 and 250 g kg⁻¹ diet) and three levels of carbohydrate (300, 340 and 380 g kg⁻¹ diet). In addition to the six experimental diets, a diet containing the protein and carbohydrate requirement levels of 300 and 260 g kg⁻¹ diet, respectively, as reported earlier for this species, was used as a reference diet. For each dietary treatment, 30 healthy fry of 20 days age (0.12 ± 0.01 g) were stocked in triplicate tanks using a flow‐through system. The fish were fed ad libitum four times a day to a level close to apparent satiation. Batch weighing of fish was done after 15 days of stocking to measure growth and general health status of the fish. The fish fed 250 g protein and 340 g carbohydrate kg⁻¹ diet with a protein to energy ratio of 17.86 g protein MJ⁻¹ performed equally well in terms of growth and nutrient utilization as the reference diet group. The study indicates that dietary protein can be reduced from 300 to 250 g kg⁻¹ diet by increasing carbohydrate from 260 to 340 g kg⁻¹ diet without sacrificing the growth of silver barb fry.     

Protein requirements of Nile tilapia (Oreochromis niloticus) fry cultured at different salinities

Effect of isolipidic (62.7 ± 5.0 g kg^?1) diets with protein levels of 204.6 (T20), 302.3 (T30), 424.6 (T40) or 511.0 g kg^?1 (T50) on growth and survival in Nile tilapia (Oreochromis niloticus Linnaeus 1758) fry cultured for 70 days at one of four salinities (0, 15, 20 and 25 g L^?1) was evaluated. A bifactorial (4 × 4) design was used with 16 treatments run in triplicate and 20 fry (0.25 ± 0.04 g) per replicate under semi‐controlled conditions. Four independent, recirculating systems (one per salinity level) were used, each one with 12 circular tanks (70 L capacity), filters and constant aeration. The different salinities had no significant effect on growth. Weight gain improved significantly as dietary protein content increased, although organisms fed the T50 diet had a lower growth rate. Survival was highest (98.33%) in the T50/15 (protein/salinity levels) treatment and lowest (71.0%) in the T20/20 treatment, with no pattern caused by the variables. The T40/25, T40/20 and T50/0 treatments produced the most efficient growth and feed utilization values while the T20 treatments at all the salinities resulted with the lowest performance. With the exception of the T50 treatments, a non‐significant tendency to increased weight gain was observed as water salinity increased, suggesting that the salinity of the culture environment does not influence dietary protein requirements in Nile tilapia O. niloticus fry.     

Effect of dietary energy content on gilthead sea bream (Sparus aurata) feeding behaviour and nutritional use of the diet

We have studied how the diet energy level affects gilthead sea bream (Sparus aurata) daily pattern of demand‐feeding activity and nutritional use of the diet under summer‐ and winter‐like conditions. To that end, animals were kept in a closed circuit under controlled temperature and photoperiod, and fed one of two commercial diets with either high (H: 238 g kg^?1 fat) or low (L: 172 g kg^?1 fat) energy content. In summer conditions (26 °C; 12 : 12 L : D), both diets yielded similar growth rate (0.7 ± 0.0 and 0.7 ± 0.1) and thermal unit growth coefficient values (0.6 ± 0.0 and 0.6 ± 0.0), and although the daily rate of delivered feed was somewhat higher for fish under the high‐energy diet (16 ± 2 g kg^?1 of fish) the difference was not significant. In winter conditions (17 °C; 9 : 15 L : D), on the other hand, no differences were found for any of the parameters evaluated. Thus, the higher lipid content of the diet does not appear to have any protein sparing effect. The ‘summer’ demand‐feeding pattern displayed three daily peaks, which were quite apparent for fish under the low‐energy diet, but rather smoothed for those that ate the energy‐rich diet. This profile was replaced in winter conditions by a single peak around noon, and the animals also displayed a lower overall number of demands.     

Tub gurnard Chelidonichthys lucerna L.: a new fish species suitable for farming? First answers evaluating the growth of juveniles reared at different stocking densities,welfare and fillet quality

A trial was conducted to evaluate the growth performance and survival of Chelidonichthys lucerna. A total of 13 352 180‐days old juveniles (5.5 ± 2 g; 5 ± 1 cm) were reared at two different densities (A‐EXP = 68 fish m^?3; B‐FFA = 15 fish m^?3) in eight tanks (four tanks per group) for 360 days. The welfare status and meat quality of fish were evaluated for the A‐EXP and B‐FFA groups in comparison with wild‐caught fishery gurnard (C‐WID). The survival rate was high for both A‐EXP (79%) and B‐FFA (93.5%). B‐FFA fish had the highest specific growth rate (1.16 vs. 1.07; P < 0.05), and were heavier than A‐EXP fish (321 ± 40 g vs .239 ± 44 g; P < 0.01). Rearing conditions did not affect blood metabolites, except for glucose concentrations, which were higher in C‐WID (P < 0.05). The meat quality traits showed that reared groups were fatter (2.8–3.2%) than C‐WID (0.94%); total n‐3 fatty acids (19.02–19.26%) were lower in reared groups than C‐WID (29.99%); and EPA + DHA were similar in all groups (15.1–16.61% vs. 27.99%). Despite the good growth and survival, the final mean weight was below that requested by the market (400–500 g). Future research efforts should focus on reducing the feed conversion rate (3:1).     

Effect of stocking density on growth performance of juvenile sea cucumber Holothuria tubulosa (Gmelin, 1788)

Stock density is one among the most important factors in aquaculture that directly influences the growth of organisms; however, there is limited information about the effects of stocking density on growth performance of sea cucumber Holothuria tubulosa (Gmelin, 1788) that has a commercial potential and represents a new species for aquaculture. In this study, we investigated the effect of stocking densities on the growth performance of juvenile H. tubulosa (40.3 ± 3.34 g) in laboratory conditions. Stocking densities were selected as 6, 15 and 30 ind m^?2 with total biomass 253.3 ± 0.18, 601.2 ± 0.11 and 1201.4 ± 0.15 g m^?2 respectively. We monitored the growth by wet weights and calculated the growth performance through weight gain, growth rate, specific growth rate, relative weight gain, coefficients of variation and survival rate during 8 weeks of research period. We found that individual mean weight gain in 6 ind m^?2 group was approximately 29.53 g while it was 3.03 g and ?4.36 g for 15 ind and 30 ind m^?2 groups respectively. Results have shown that the specific growth rate tends to decrease as the number of individuals in unit area increases. The final mean weight, growth rate, specific growth rate, relative weight gain and coefficients of variation were significantly different among treatment groups. In conclusion, a density of 6 ind m^?2 is recommended for stocking juvenile H. tubulosa under rearing conditions. Stocking density of 15 ind m^?2 is not favourable for long term as the growth rate is negligible where 30 ind m^?2 should be avoided in tank‐based rearing units.     

Growth performance of the white shrimp,Litopenaeus vannamei,fed on practical diets with increasing levels of the Antarctic krill meal,Euphausia superba,reared in clear‐ versus green‐water culture tanks

Litopenaeus vannamei were stocked in 25 clear‐water 500‐L tanks at 100 shrimp m^?2 and in 25 green‐water 1000‐L tanks at 60 animals m^?2. Four diets were formulated to include krill meal at 10, 50 or 110 g kg^?1; or krill oil at 25 g kg^?1 by replacing fish meal, fish oil, soybean lecithin and cholesterol. Diets had similar levels of crude protein, total energy and essential amino acids. After 72 days, shrimp reared in clear and green water showed no differences in performance among diets. In clear water, shrimp attained 13.1 ± 0.59 g body weight, 1.00 ± 0.06 g week^?1 growth, 81.4 ± 7.3% survival, 780 ± 118 g m^?2 yield, 16.9 ± 1.8 g shrimp^?1 apparent feed intake (AFI), and 2.18 ± 0.29 food conversion ratio (FCR). In green water, shrimp attained 14.3 ± 0.81 g body weight, 1.04 ± 0.09 g week^?1 growth, 91.4 ± 5.4% survival, 569 ± 69 g m^?2 yield, 20.9 ± 3.2 g shrimp^?1 AFI, and 2.22 ± 0.34 FCR. Diets containing krill meal or krill oil were able to fully replace the protein and lipid value of fish meal, fish oil, soybean lecithin and cholesterol at no cost to performance.     

Response of the tropical spiny lobster Panulirus ornatus to protein content of pelleted feed and to a diet of mussel flesh

In an 8‐week growth experiment, juvenile spiny lobsters (Panulirus ornatus) grew best on a feed containing at least 610 g kg^?1 crude protein on a dry matter basis (DM) and a digestible protein to digestible energy ratio of 29.8 mg kJ^?1. The study entailed a six treatment by four replicate randomized block experiment with 222 wild‐caught P. ornatus of mean initial weight (±SD) of 2.5 ± 0.19 g. The lobsters were fed one of five isolipidic feeds (approximately 130 g kg^?1 DM) in which the crude protein was serially incremented between 330 and 610 g kg^?1 DM, or a reference diet comprising the flesh of frozen green‐lip mussels. Lobsters fed the pelleted feeds had high survival (79 ± 4.5%) and responded to increasing dietary crude protein content with progressively higher growth rates, with the daily growth coefficient improving from 0.72% day^?1 with 330 g kg^?1 crude protein to 1.38% day^?1 with 610 g kg^?1 crude protein. Both growth rate and survival were low with the mussel diet (0.80% day^?1and 41 ± 4.5%, respectively). These results demonstrate that tropical spiny lobsters grow well when fed high‐protein, high lipid, pelleted feeds, but feeding on a sole diet of freshly thawed green‐lip mussels was unsatisfactory.     

Supplementation with 2‐hydroxy‐4‐(methylthio)butanoic acid (HMTBa) in low fish meal diets for the white shrimp,Litopenaeus vannamei

This work evaluated the performance of Litopenaeus vannamei to low fish meal diets supplemented with 2‐hydroxy‐4‐(methylthio)butanoic acid (HMTBa). A basal diet with 150.0 g kg^?1 of anchovy fish meal was designed. Two positive control diets were formulated to reduce fish meal at 50% and 100% with 1.0 and 2.0 g kg^?1 of MERA? Met_Ca (calcium salt with 84% HMTBa activity), respectively. Two nearly equivalent diets acted as negative controls, without HMTBa supplementation. A total of 50 clear‐water tanks of 500 L were stocked with 2.22 ± 0.19 g shrimp under 70 animals m^?2. Shrimp survival (92.3 ± 5.1% and 81.4 ± 8.0%), yield (808 ± 12 and 946 ± 17 g m^?2) and FCR (2.17 ± 0.19 and 3.12 ± 0.37) showed no differences among diets after 72 or 96 days, respectively. A significantly higher shrimp body weight and weekly growth were observed for those fed with the basal diet or diets supplemented with HMTBa compared with non‐supplemented ones. This study has shown that L. vannamei growth, body weight, survival, yield and FCR were supported by HMTBa supplementation when 150.0 g kg^?1 of fish meal was replaced by soybean meal and other ingredients, at 50% and 100%.     

Leucocyte profile and growth rates as indicators of crowding stress in pejerrey fingerlings (Odontesthes bonariensis)

Crowding is one of the most common stressors found in intensive aquaculture, compromising growth rates and immune function. Plasmatic cortisol is a classic stress biomarker for fish, but its quantification is expensive and demands blood volumes that small individuals do not provide, constraining the usage of this technique to assess stress in fingerlings. The leucocyte profile is an alternative methodology to quantify stress with reduced costs and volumes of blood. Stress conditions promote neutrophilia and lymphocytopenia as response to elevated glucocorticoids levels. Considering the difficulties to assess stress imposed by intensive fish farming using measurement of glucocorticoid hormones, this study aimed to evaluate the stress‐induced changes in leucocyte profiles and growth rates imposed by crowding in fingerlings of Odontesthes bonariensis, a promising South‐American candidate for freshwater aquaculture. To meet these objectives, fingerlings (initial weight 0.05 ± 0.06 g and length 1.68 ± 0.13 cm) were reared for 45 days under three rearing densities (1, 5 and 10 fingerlings L^?1). At the end of this period, fish were anaesthetized and euthanized to obtain the leucocyte profile, neutrophil:lymphocyte ratio (N:L) and growth rate. Increasing density promoted: significant reduction in growth (final length, weight and specific growth rate); neutrophilia and lymphocytopenia; and increased N:L ratio. Concluding, the tested rearing densities imposed distinct levels of stress characterized by different N:L ratio, demonstrating that the leucocyte profile is a reliable alternative to measure stress levels in O. bonariensis fingerlings and probably in other fish species.