Optimal dietary protein requirement for juvenile ivory shell, Babylonia areolate

A growth experiment was conducted to determine the optimal dietary protein requirement for juvenile ivory shell reared in indoor aerated aquaria. Six isoenergetic experimental diets using fish meal, casein and gelatin as protein sources were formulated to contain graded levels of protein (27, 33, 38, 43, 49 and 54% of dry diet, respectively). Triplicate groups of 40 shells (average weight 93.50 ± 1.70 mg) were stocked in 120-l tanks and fed to apparent satiation twice daily for 8 weeks. The results showed that the growth performance and feed utilization were significantly affected by dietary protein level (P < 0.05). Maximum weight gain, mean protein gain, specific growth rate and soft body to shell ratio occurred at 43% dietary protein level (P < 0.05). There were significant differences in protein, lipid, moisture and ash content in soft body; except that ash content in shell was not significantly affected by dietary protein level. Pepsin activity in soft body tissue significantly increased with dietary protein level up to 43%, and trypsin-like enzyme activity increased with dietary protein level up to 49%. However, lipase activity in soft body decreased with increasing dietary protein level. However, no significant differences (P < 0.05) in survival, calcium, phosphorus concentration in the shell and soft body were found among dietary treatments. Quadratic regression analysis of weight gain against dietary protein level indicated that the optimal dietary protein requirement for maximum growth and feed utilization of juvenile ivory shell is 45% of dry diet.     

The minimum dietary lysine requirement,maintenance requirement and efficiency of lysine utilization for growth of Atlantic salmon smolts

This study was conducted to evaluate and refine the dietary lysine requirement of Atlantic salmon (Salmo salar) smolts using both the dose–response technique to determine the minimum dietary requirement and different ration levels to determine the maintenance lysine requirement and the efficiency of lysine utilization for growth. Diets containing 1.97–3.16% lysine (presented in protein‐bound form of lysine) were fed to satiation or at 25–75% ration levels to smolts during the last week in fresh water and the following 9 weeks in seawater. Based on the exponential lysine gain response to increasing dietary lysine concentrations, the minimum lysine requirement was found to be 2.67% of diet (dry matter basis; protein‐bound form or 3.05% free form). The smolts fed the unsupplemented diet maximally utilized the digestible lysine intake for growth. The efficiency of digestible lysine utilization for growth was 0.85 in the diet containing 2.88% lysine and the maintenance requirement was 21.7 mg digestible lysine kg^?0.7 day^?1.     

Influence of temperature on protein utilization in juvenile European seabass (Dicentrarchus labrax)

The aim of this trial was to study the utilization of dietary protein by seabass juveniles with 5.5 g mean body weight, at two water temperatures: 18°C and 25°C. For that purpose, the fish were fed for 12 weeks, four isoenergetic diets with different protein levels (36, 42, 48, and 56%). At the end of the trial, growth rate and feed utilization were significantly better at the higher water temperature. Within each temperature, specific growth rate and feed efficiency were significantly higher with the 48 and 56% protein diets than with the other diets. At 25°C, feed efficiency was also significantly better with the 56% than with the 48% protein diet. N retention (g kg average body weight⁻¹ day⁻¹) was higher at 25°C than at 18°C but, as a % N intake the inverse was true. Although at 25°C N retention (% N intake) was not different among groups, retention in g kg ABW⁻¹ day⁻¹ was significantly higher with the 56% protein diet than with 36 and 42% protein diets. On the contrary, at 18°C N retention (g kg ABW⁻¹ day⁻¹) was similar among groups while as a percentage of N intake it was inversely related to the dietary protein level. Regarding energy utilization, at each temperature, there were no differences among dietary treatments on energy retention (g kg ABW⁻¹ day⁻¹). As a % of energy intake, energy retention significantly increased with the increase of dietary protein level at 25°C, while at 18°C, there were no significant differences among groups. Within each temperature, at the end of the trial, there were no differences among groups in proximate composition of whole fish. Apparent digestibility coefficients of dry matter, protein and energy significantly improved with the increase of water temperature but, within each temperature, there were no significant differences among groups. The results of this study indicate that, regardless of water temperature, the dietary protein requirement for growth seems to be satisfied with a diet containing 48% protein. Growth and feed efficiency were significantly higher at the higher temperature, however, protein utilization was more efficient at the lower temperature.     

Energy and nitrogen partitioning in 250 g Atlantic cod (Gadus morhua L.) given graded levels of feed with different protein and lipid content

The effects of feed intake level on energy and nitrogen partitioning were studied in juvenile Atlantic cod (250 g) fed two fish meal based diets differing in protein and lipid content (54:31 and 65:16) at 10 °C. Replicate groups of cod were feed deprived for 32 days or fed one of the two diets at 25, 50, 75 or 100% of group satiation for 60 days. Feed intake and oxygen consumption were measured daily and weights and chemical composition of carcass, liver, viscera and whole body were measured at start and end. Diet digestibilities were assessed in a separate experiment.

The whole body and carcass growth rates at a given feed intake did not differ between dietary groups, but the liver grew faster in the fish fed the low protein diet, resulting in higher hepatosomatic indices at the end of the experiment in the groups fed this diet.

The efficiency of utilisation of digestible nitrogen for growth (k_DNg) was higher for the low protein diet (0.73 ± 0.02) than for the high protein diet (0.53 ± 0.05), resulting in higher nitrogen retention at a given nitrogen intake. No difference in percentage nitrogen retention was seen in full-fed fish however (31.2 ± 2.5 and 28.4 ± 1.6% for the low protein and high protein diets, respectively). This can be explained by higher nitrogen intake in the fish fed the high protein diet, resulting in a smaller proportion of the intake being used for maintenance.

There was no difference in energy utilisation between dietary groups. The digestible energy requirement for maintenance (DE_maint) was 53.8 ± 0.9 kJ kg^− 1 d^− 1 (42.3 ± 0.7 kJ kg^− 0.8 d^− 1) and the utilisation efficiency for growth (k_DEg) was 0.80 ± 0.02. The energy retention in full-fed fish was 31.3 ± 3.5 and 31.7 ± 1.0% for the low protein and high protein diets, respectively. The deposited energy was distributed in approximately equal proportions in the liver and carcass, whereas viscera accounted for a minor proportion. At a given energy intake, the fish fed the high protein diet deposited more energy in the carcass and less in the liver than did those fed the low protein diet.     

Dietary l‐lysine requirement of fingerling stinging catfish,Heteropneustes fossilis (Bloch) for optimizing growth,feed conversion,protein and lysine deposition

Dietary lysine requirement of fingerling Heteropneustes fossilis (6.96 ± 0.05 g) was quantified by conducting 12‐week feeding trial in a flow‐through system at 28°C. Casein–gelatin based isonitrogenous (38% CP) and isocaloric (14.7 kJ g^?1 DE) amino acid test diets with six levels of dietary lysine (1.5%, 1.75%, 2.0%, 2.25%, 2.5%, 3.0% dry diet) were fed to apparent satiation in triplicates. Broken‐line and second‐degree polynomial regression analyses at 95% plateau of absolute weight gain (AWG; g fish^?1), feed conversion ratio (FCR), protein deposition (PD; g fish^?1) and lysine deposition (LD; g fish^?1) exhibited lysine requirement between 2.0% to 2.3% of the dry diet, corresponding to 5.3–6.1% protein.     

Application of different types of protein hydrolysate in high plant protein diets for juvenile turbot (Scophthalmus maximus)

A 10‐week feeding experiment was conducted to investigate the effects of different types of dietary protein hydrolysate (PH) on growth performance, body composition, trypsin activity and serum transaminase of juvenile turbot. Four high plant protein diets contained different types of PH, fish PH (FPH), yeast PH (YPH), pig blood PH (PBPH) and soy PH (SPH), replacing 10% fishmeal of the basal diet. The basal diet with 30% fishmeal and no PH was used as the control diet (C). Each diet was assigned to triplicate groups of 30 fish. The specific growth rate (SGR) was not significantly different between groups C and FPH, but groups C and FPH showed significantly higher SGR than other groups. The feed efficiency ratio (FER) and protein efficiency ratio (PER) were not significantly different among groups C, FPH and YPH, but groups PBPH and SPH showed significantly lower FER and PER than groups C and FPH. Group PBPH showed significantly higher hepatosomatic index than other groups except YPH. Fish fed YPH showed significantly lower whole‐body protein content, but significantly higher whole‐body lipid content than fish fed diets C, FPH and PBPH. The activities of serum GOT and GPT in group PBPH were higher than those in groups C, FPH and YPH. These results suggested that when used at a low level in high plant protein diets for juvenile turbot, FPH is a good alternative protein source and YPH also has the application potential, but PBPH and SPH can cause negative impacts on fish growth and health.     

Effects of dietary levels of protein on growth,feed utilization and physiological parameters for juvenile Dabry's sturgeon,Acipenser dabryanus

Five semi‐purified diets were formulated to contain graded levels of dietary protein (340, 390, 440, 490 and 540 g/kg diet), and each diet was assigned to triplicate groups of eight juvenile Dabry's sturgeon for 50 days. Results showed that specific growth rate increased linearly with increasing dietary protein levels from 340 to 440 g/kg diet, then levelled off at higher protein levels. Higher dietary protein (440 and 540 g/kg) significantly increased the feed intake and feed efficiency ratio of fish compared to lower protein diets (340 to 490 g/kg diet) (p < .05). Fish fed a 440 g protein kg^?1 diet had higher protein efficiency ratio and serum lysozyme activity than other treatments. Serum ammonia content and activities of liver aminotransferase were positively correlated with dietary protein levels. No significant difference (p > .05) among groups was observed in glycogen content. As dietary protein level increased, protein and ash content of fish dorsal muscle were greatly enhanced, whereas lipid content was significantly reduced (p < .05). Based on broken‐line regression analysis of SGR against dietary protein levels, the optimum dietary protein level for maximal growth of juvenile Dabry's sturgeon is 446.68 g/kg diet.     

The effect of dietary protein replacement by crystalline amino acid on growth and nitrogen utilization of turbot Scophthalmus maximus juveniles

A study was conducted to evaluate the effect of partial replacement of dietary fish meal by crystalline amino acids on growth performance, feed utilization, body composition and nitrogen utilization of turbot juveniles.

Four diets were formulated to be isolipidic (12% DM) and isonitrogenous (8% DM). A fish meal based diet was used as control. In the experimental diets, a crystalline amino acid (AA) mixture was used to partially replace fish meal, corresponding to a non-protein nitrogen content of 19, 37 and 56%, respectively (diets 19AA, 37AA and 56AA, respectively). The overall amino acid profile of the experimental diets resembled that of the whole-body protein of turbot. Each experimental diet was fed to triplicate groups of 20 fish (initial body weight of 31.8 g) twice daily to apparent satiation for 42 days. During the trial water temperature averaged 18 °C.

Final body weight, weight gain (g kg ABW^− 1 day^− 1) and specific growth rate were not different between the control and 19AA diet but significantly decreased with the increase of crystalline-AA inclusion from 19 to 56%. Feed intake and feed efficiency of fish fed the control and diet 19AA were similar and significantly higher than those of fish fed the 56AA diet. At the end of the growth trial, there were no significant differences in whole-body composition among groups. Hepatosomatic index was also unaffected by dietary treatments.

Nitrogen retention (g kg ABW^− 1 day^− 1) of fish fed the control and the 19AA diets were similar and significantly higher than that of fish fed the other diets. Expressed as a percentage of the nitrogen intake, N retention was significantly higher with the control than with the 37AA and 56AA diets.

Daily ammonia excretion (mg kg ABW^− 1 day^− 1) of fish fed the control diet was significantly higher than that of fish fed the 37AA and 56AA diets, while daily urea excretion (mg kg ABW^− 1 day^− 1) did not significantly differ among treatments. Non-fecal nitrogen (ammonia + urea) excretion (mg kg ABW^− 1 day^− 1) was significantly higher for fish fed the control diet than in those fed the 37AA and 56AA diets. However, as percent of N intake, ammonia excretion and non-fecal N excretion were significantly higher in fish fed the 56AA diet than in those fed the control and 19AA diets.

Specific activity of glutamate dehydrogenase, alanine and aspartate aminotransferases did not significantly differ among experimental groups.

In conclusion, in diets with an overall amino acid profile resembling that of the whole-body protein of turbot, crystalline-AA may replace 19% of dietary protein without negatively affecting growth performances or feed utilization efficiency. However, higher protein replacement levels of protein-bound-AA by crystalline-AA severely depressed growth performance.     

Effects of replacement of animal protein by soybean meal on growth and carcass composition in juvenile Australian freshwater crayfish

A feed trial was conducted for 59 days with juvenile Cherax destructor, mean weight (se) 0.61 (0.01) g, reared communally and maintained on 16 isoenergetic diets containing crude protein levels of 15, 20, 25, and 30%. For each protein level the fish meal component was replaced by soybean meal to produce diets in which 0, 20, 40, or 60% of the protein originated from soybean meal. Mean percentage weight gain per day ranged from 2.98% (15% protein, 60% soybean meal diet), to 11.75% (30% protein, 40% soybean meal diet). When soybean meal was included at a level of 40–60%, growth rate was reduced relative to that achieved with control diets at 15% and 20% protein levels. In no case did a 20% substitution significantly affect growth over that achieved with controls. A two-way interaction occurred between dietary protein and the level of dietary soybean meal. Feeds of higher protein content appeared to permit higher soybean meal inclusion levels without significantly affecting growth. Increases of 5% protein produced a significant improvement in growth when soybean meal contributed from 40–60% of the total protein. This effect was less pronounced in the control diets and the 20% soybean meal series. The percentages of protein increased and lipid decreased in the carcass as the level of dietary protein increased. A similar effect occurred by increasing the soybean meal substitution level to 60%. An obvious trend in carcass moisture, energy, and ash did not occur. A protein requirement of 30% is apparent when fish meal and soybean meal are included in diets at levels of 20% and 24% respectively. A maximum weight of 14.13 g was recorded for an individual fed the 30% protein, 20% soybean meal diet.     

The effect of dietary protein source on growth and carcass composition in juvenile Australian freshwater crayfish

A feed trial was conducted for 12 weeks on juvenile Australian freshwater crayfish (Cherax destructor) (mean weight (SE) 0.82 (0.02)g) maintained on five isoenergetic diets with a protein content of 30%. Diets differed in the primary source of protein used, with meat, snail, soybean, yabby, and zooplankton meals comprising the major protein ingredient, varying from 56–60% of total protein. Mean percentage weight gain per day ranged from 7.57% (yabby meal diet), to 9.42% (snail meal diet). No significant difference occurred in mean weight, percentage weight gain, specific growth rate (%), or survival among diets. A maximum size of 16.44g was attained on the yabby meal diet. Largest mean weight was 8.27g on the snail-based diet. Food conversion ratios were all good, with a minimum value of 0.95 recorded for the snail-based diet. Initial food consumption per day was approximately 5% of mean animal weight, decreasing to around 2.4%, and is collectively described by a power curve. Protein retention ranged from 29.57% in the zooplankton meal diet to 41.15% in the snail-based diet. Carcass composition was influenced by feed type, with the most marked difference occurring in carapace colour. Animals fed the zooplankton-based pellets developed the strongest pigmentation. Recommendations are made for including certain protein-based ingredients in manufactured yabby diets.     

Dietary threonine requirement of fingerling Indian major carp,Catla catla (Hamilton) estimated by growth,protein retention efficiency,threonine deposition,haematological parameters and carcass composition

A 12‐week feeding trial was conducted to determine the dietary threonine requirement of fingerling Indian major carp, Catla catla (3.35 ± 0.11 cm; 0.59 ± 0.06 g). Six casein‐gelatin based (33% crude protein; 3.23 kcal g^?1 digestible energy) amino acid test diets with graded levels of analysed threonine (0.74%, 0.96%, 1.21%, 1.48%, 1.72% and 1.93% dry diet) were fed to satiation to triplicate groups of fish. Absolute weight gain (g per fish), feed conversion ratio, protein retention efficiency, threonine deposition, RNA/DNA ratio and carcass protein significantly improved with the increase in dietary threonine and peaked at 1.48% of the dry diet. Haematological indices were also found to be best in fish fed at 1.48% threonine diet. Quadratic regression analysis of absolute weight gain, feed conversion ratio, protein retention efficiency, threonine deposition, RNA/DNA ratio, carcass protein, haemoglobin (g dL^?1), haematocrit (%) and RBCs (10⁶ × mm^?3) at 95% of maximum and minimum response exhibited the threonine requirement of fingerling C. catla between 1.35% and 1.48% dry diet, corresponding to 4.09–4.48% dietary protein. Present finding would be useful in formulating threonine‐balanced feeds for the intensive culture of C. catla.     

Effect of ration size on growth,body composition,and energy and protein maintenance requirement of fingerling Indian major carp, <Emphasis Type="Italic">Labeo rohita</Emphasis> (Hamilton)

An eight-week feeding trial has been conducted to determine the optimum ration for Indian major carp, Labeo rohita, fingerling (4.10 ± 0.30 cm, 0.55 ± 0.16 g) by feeding a purified diet (40% CP; 3.61 kcal g⁻¹ GE) at six levels, 2, 4, 6, 8, 10, and 12% of body weight per day, at 0800 and 1600 h, in triplicate, to 20 fish per trough fitted with a water flow-through system. Highest weight gain, best feed conversion ratio (FCR), best specific growth rate (SGR%), and highest protein efficiency ratio (PER) were evident for rations of 6–8% body weight. Second-degree polynomial regression analysis for FCR, PER, protein, and energy retention data indicated the break-points occurred at 6.55, 6.75, 6.80, and 6.95% bw per day, respectively. Significant (P < 0.05) differences between body composition were observed for fish fed different rations. Maximum body protein content was recorded for 6% and 8% rations. A linear increase in body fat content was evident with increasing ration. Body moisture and ash content remained non-significantly (P > 0.05) low for higher rations, however. On the basis of these results it is recommended that feeding in the range 6.5–7.0% bw per day corresponding to 2.6–2.8 g protein and 23.49–25.31 kcal energy per 100 g of the diet per day is optimum for growth and efficient feed utilization of Labeo rohita. Results for 2–4% rations (0.8–1.6 g protein and 7.23–14.46 kcal energy) suggest these amounts approximate to the maintenance requirement of this fish.     

Effect of protein levels on growth, feed utilization, nitrogen and energy budget in juvenile southern flounder, Paralichthys lethostigma

A feeding trial with five crude protein levels (549, 513, 472, 445 and 399 g kg^?1) was conducted to investigate the optimum protein level in diets of juvenile southern flounder (Paralichthys lethostigma). Budgets of nitrogen and energy were discussed. Fish (initial weight 32.9 ± 0.5 g fish^?1, mean ± SD) were fed the experimental diets to satiation twice daily for 61 days. Protein levels affected specific growth rate in wet weight (SGR_W) and protein (SGR_P) significantly. SGR_W and SGR_P were highest at 512.5 g kg^?1 protein level. SGR_W was positively correlated to growth nitrogen (GN), growth energy (GE), nitrogen digestibility, energy digestibility, amount of digestible nitrogen and amount of digestible energy. Faecal nitrogen (FN) and faecal energy (FE) were affected significantly with trends contrary to SGR_W. The nitrogen budget was described by the equation 100CN = 2.1FN + 34.4UN + 63.5GN (CN, nitrogen intake; UN, excretion nitrogen). The energy budget was 100IE = 4.04FE +3.32UE + 54.35GE + 38.30ME (IE, gross energy intake; UE, excretion energy; ME, metabolizable energy). The average proportion of GE and ME in assimilated energy (AE) was described by the equation 100AE = 58.65GE + 41.35ME.     

Nucleotide promotes feed intake and protein utilization via regulating the gene expression of feeding and nitrogen metabolism in juvenile Chinese perch (Siniperca chuatsi)

Nucleotide (NT) could enhance growth, feeding and immunity in higher vertebrate and fish. Chinese perch (Siniperca chuatsi) refuse artificial diet in natural water, whereas the NT might promote its feeding of domestication diet. The present study was designed to investigate the effect of dietary NT on growth, feed intake, feed utilization, body composition, serum biochemistry and nitrogen metabolism in juvenile Siniperca chuatsi. 0, 0.5, 1.0, 1.5, 2.0 and 4.0 g NT kg^?1 diet were supplemented in each diet, respectively. Triplicate groups of Chinese perch (36.21 ± 0.98 g) were fed twice a day to apparent satiation for 8 weeks. The results showed that fish fed with 1.5 g NT kg^?1 diet had the highest weight gain (WG), specific growth rate, feed efficiency, feed intake, protein efficiency ratio and protein retention efficiency. The crude protein content in whole body was highest in fish fed the diet containing 1.5 g NT kg^?1 diet. Dietary NT supplementation significantly increased the neuropeptide Y gene expression and significantly decreased the agouti‐related protein and pro‐opiomelanocortin gene expressions in brain. NT supplementation decreased urea nitrogen content, aspartate aminotransferase (AST) and alanine aminotransferase activities in serum. The hepatic AST activity was increased first and then decreased, and the highest activity was observed in fish fed with 1 g NT kg^?1 diet. Inversely, the activity of glutamate dehydrogenase (GDH) in liver and adenosine 5′‐monophosphate deaminase in muscle decreased first and then increased, and the lowest activity was observed in fish fed with 1.5 g NT kg^?1 diet. Similarly, the gene expression levels of GDH and arginase in liver were lowest in fish fed with 1.5 g NT kg^?1 diet. Based on the broken‐line regression analysis of WG in the present experimental condition, 1.44 g NT kg^?1 diet was the optimum supplementation level in juvenile Siniperca chuatsi.     

Prediction of apparent protein digestibility of ingredients and diets by in vitro pH-stat degree of protein hydrolysis with species-specific enzymes for juvenile Pacific white shrimp Litopenaeus vannamei

Aquafeed production faces global issues related to availability of feed ingredients. Feed manufacturers require greater flexibility in order to develop nutritional and cost-effective formulations that take into account nutrient content and availability of ingredients. The search for appropriate ingredients requires detailed screening of their potential nutritional value and variability at the industrial level. In vitro digestion of feedstuffs by enzymes extracted from the target species has been correlated with apparent protein digestibility (APD) in fish and shrimp species. The present study verified the relationship between APD and in vitro degree of protein hydrolysis (DH) with Litopenaeus vannamei hepatopancreas enzymes in several different ingredients (n = 26): blood meals, casein, corn gluten meal, crab meal, distiller's dried grains with solubles, feather meal, fish meals, gelatin, krill meals, poultry by-product meal, soybean meals, squid meals and wheat gluten. The relationship between APD and DH was further verified in diets formulated with these ingredients at 30% inclusion into a reference diet. APD was determined in vivo (30.1 ± 0.5 °C, 32.2 ± 0.4‰) with juvenile L. vannamei (9 to 12 g) after placement of test ingredients into a reference diet (35 g kg^− 1 CP; 8.03 g kg^− 1 lipid; 2.01 kcal g^− 1) with chromic oxide as the inert marker. In vitro DH was assessed in ingredients and diets with standardized hepatopancreas enzymes extracted from pond-reared shrimp. The DH of ingredients was determined under different assay conditions to check for the most suitable in vitro protocol for APD prediction: different batches of enzyme extracts (HPf5 or HPf6), temperatures (25 or 30 °C) and enzyme activity (azocasein): crude protein ratios (4 U: 80 mg CP or 4 U: 40 mg CP). DH was not affected by ingredient proximate composition. APD was significantly correlated to DH in regressions considering either ingredients or diets. The relationships between APD and DH of the ingredients could be suitably adjusted to a Rational Function (y = (a + bx)/(1 + cx + dx²), n = 26. Best in vitro APD predictions were obtained at 25 °C, 4 U: 80 mg CP both for ingredients (R² = 0.86; P = 0.001) and test diets (R² = 0.96; P = 0.007). The regression model including all 26 ingredients generated higher prediction residuals (i.e., predicted APD − determined APD) for corn gluten meal, feather meal, poultry by-product meal and krill flour. The remaining test ingredients presented mean prediction residuals of 3.5 points. A model including only ingredients with APD > 80% showed higher prediction precision (R² = 0.98; P = 0.000004; n = 20) with average residual of 1.8 points. Predictive models including only ingredients from the same origin (e.g., marine-based, R² = 0.98; P = 0.033) also displayed low residuals. Since in vitro techniques have been usually validated through regressions against in vivo APD, the DH predictive capacity may depend on the consistency of the in vivo methodology. Regressions between APD and DH suggested a close relationship between peptide bond breakage by hepatopancreas digestive proteases and the apparent nitrogen assimilation in shrimp, and this may be a useful tool to provide rapid nutritional information.     

Replacing fish meal with vegetable protein sources in feed for juvenile red swamp crayfish,Procambarus clarkii: Effects of amino acids supplementation on growth and feed utilization

An 8‐week growth trial was conducted to investigate the effects of dietary fish meal replacement with a vegetable mixture of soybean meal and rapeseed meal (1:1) on growth of juvenile red swamp crayfish. Nine isonitrogenous diets were designed: V0, V34, V50, V65, V73 and V81 with six levels of vegetable proteins, and VA48, VA63 and VA78 by further adding crystalline lysine and methionine into V50, V65 and V81. Compared with V0, V34 significantly improved the specific growth rate (SGR), while V65, V73, V81 and VA78 depressed the SGR (p < .05). Feeding rate showed a decreasing trend as dietary vegetable protein level increased (p < .05), except that in VA48 group. Significantly lower FCR and higher PER were observed in V34 group, whereas all vegetable protein diets depressed the feed utilization of crayfish (p < .05). Crayfish fed with diets containing vegetable proteins showed significantly lower hepatosomatic indices and higher condition factors than the control (p < .05). Muscle lipid content was significantly (p < .05) lowered in V81 group, but not in VA78 group. The results suggested that 338 g/kg vegetable protein improved growth performance of crayfish. Excessive vegetable protein depressed the growth of crayfish, which could be prevented by lysine and methionine supplementation except for the all vegetable protein diets.     

Development of a black sea bream fibroblast cell line and its potential use as an <Emphasis Type="Italic">in vitro</Emphasis> model for stress protein studies

A fibroblast cell line (BSF) derived from a caudal fin explant of black sea bream (Mylio macrocephalus) was developed. The optimum fetal bovine serum (FBS) concentration for fibroblast cell line growth was found to be 15–20% v/v FBS and the optimum temperature range for growth was found to be 26–30 °C. The fibroblast cells displayed a diverse distribution in chromosome number with two modal chromosome numbers of 48 and 54. Upon acute heat shock (+8 °C) the cells displayed a 4.1 fold increase in hsp70 and this elevation was not prolonged as hsp70 returned to near basal levels following a 6 h recovery period. The effect of the hsp70 inducer L-azetidine- 2-carboxylic acid was tested and it was found that at a concentration of 10 mM this inducer caused a 2.3 fold increase in hsp70 levels. The sensitivity of the fibroblast cell line to heavy metal exposure was tested by treatment with Cu2+ and it was found that hsp70 was significantly elevated in the presence of micromolar concentrations of Cu2+. The data from this study demonstrates that the established black sea bream fibroblast cell line could serve as a useful in vitro model for stress protein studies.     

Replacement of fish meal by potato protein concentrate in diets for rainbow trout, Oncorhynchus mykiss (Walbaum): growth, feed utilization and body composition

Five experimental diets (PPC1, PPC2, PPC3, PPC4 and PPC5) containing about 465 g kg⁻¹ crude protein were formulated to contain graded levels of potato protein concentrate (PPC) (a by-product of potato starch manufacturing) by replacement of fish meal protein. The contents of PPC were 0, 94, 200, 304 and 510 g kg⁻¹ respectively. The diets were prepared isonitrogenously and isocalorically. A 6-week growth trial was conducted with rainbow trout, Oncorhynchus mykiss (Walbaum), at about 14°C to evaluate the potential use of the potato protein concentrate in trout diets.
The results showed that fish fed diets containing PPC showed decreased growth, feed utilization and condition factor, and increased mortality when PPC constituted the sole protein source. Incorporation of PPC in the diets resulted in decreased dry matter, protein and fat and increased ash contents of the fish body. In conclusion, the type of potato protein concentrate used, especially at high levels, was not suitable as an ingredient in trout feed.     

Induction of apoptosis in sea bream fibroblasts by Vibrio harveyi haemolysin and evidence for an anti‐apoptotic role of heat shock protein 70

In this study, we exposed black sea bream, Mylio macrocephalus (Basilewsky), fibroblast (BSF) and silver sea bream, Sparus sarba Forsskål, fibroblast (SSF) cell lines to a recombinant Vibrio harveyi haemolysin (VHH) and investigated mechanisms involved in apoptosis. A decrease in mitochondrial membrane potential, followed by an increase in caspase 3 activity, occurred within 2–8 h of VHH exposure, in both cell lines; however, VHH did not alter cellular levels of reactive oxygen species. As heat shock protein 70 (HSP70) is known to prevent the onset of apoptosis in certain mammalian cells, we aimed to test whether such a protective effect is operative in VHH‐exposed fibroblasts. The amounts of HSP70 were elevated in SSF and BSF via an acute heat shock or an acute heat shock followed by a 6 h recovery. It was found that the VHH‐mediated reduction in mitochondrial membrane potential was suppressed in cells that had a 6 h post‐heat shock recovery, and the protective effect of heat shock‐induced HSP70 was attenuated following treatment of cells with the HSP70 inhibitor, quercetin. This study demonstrates how haemolysin causes cell death via induction of apoptosis and provides evidence as to the role of HSP70 as an anti‐apoptotic factor.