Effect of dietary chitosan on growth performance,hematological parameters,intestinal histology and stress resistance of Caspian kutum (<Emphasis Type="Italic">Rutilus frisii kutum</Emphasis> Kamenskii, 1901) fingerlings

This study was conducted to investigate the effects of dietary chitosan on growth performance, hematological parameters, intestinal histology, stress resistance and body composition in the Caspian kutum (Rutilus frisii kutum, Kamenskii, 1901) fingerlings. Fish (1.7 ± 0.15 g) were fed diets containing chitosan at different levels (0, 0.25, 0.5, 1 and 2 g kg^?1 diet) for a period of 60 days. Results showed that the feed conversion ratio significantly decreased in fish fed diet containing 1 g kg^?1 of chitosan compared to the other groups (P < 0.05), but there were no significant differences between treatments in terms of specific growth rate and condition factor (P > 0.05). Leukocyte increased in fish fed diet containing 2 g kg^?1 of chitosan compared to the other groups (P < 0.05). Lymphocytes, eosinophils and neutrophils did not significantly change among dietary treatments (P > 0.05). Also, the chitosan supplementation did not affect the whole-fish body composition (P > 0.05). Light microscopy demonstrated that the intestinal villus length increased in fish fed diet containing 1 g kg^?1 of chitosan compared to control group (P < 0.05). While 11 and 13 ‰ salinity and 30 °C thermal stress had no effect, 1 g kg^?1 of chitosan (P < 0.05) showed highest survival rate (70 %) in 34 °C thermal stress. The results showed that chitosan in the diet of the Caspian kutum fingerlings could improve feed conversion ratio, the nonspecific defense mechanisms and resistance to some of the environmental stresses.     

Growth performance and biochemical composition of juvenile pearl oyster <Emphasis Type="Italic">Pinctada martensii</Emphasis> fed on artificial diets

The effects of three artificial diets (S1, S2 and S3) on survival, growth and biochemical composition of one-year-old pearl oyster Pinctada martensii were investigated. Six experimental groups (EG1, EG2, EG3, EG4, EG5 and EG6) and one control group (CG) were set up. EG1, EG2 and EG3 were solely fed on S1, S2 and S3, respectively. EG4, EG5 and EG6 were fed on mixed diets, as follows: S1 and Platymonas subcordiformis; S2 and P. subcordiformis; and S3 and P. subcordiformis, respectively. CG was fed on only P. subcordiformis. All groups were continuously fed for 60 days. Survival, growth and biochemical composition of soft tissues were compared across the groups. Results showed that survival rate, the absolute growth rate (AGR) and relative growth rate (RGR) of shell length did not differ significantly across the groups (p > 0.05). The AGR and RGR of total weight differed significantly among the groups (p < 0.05). AGR and RGR of shell length and total weight were the highest in EG5 and the lowest in EG1. Gross fat content showed insignificant differences among the groups (p > 0.05). However, gross protein content and ash content showed significant differences across the groups (p < 0.05). The gross protein of the groups solely fed on artificial diets was lower than those of the groups fed on mixtures of artificial diets and microalgae or single microalgae. The contents of other amino acids, total amino acids (TAAs) and essential amino acids (EAAs) showed significant differences across the groups (p < 0.05). TAA, EAA and delicious amino acids of the groups fed solely on artificial diets were lower than those of the groups fed on mixtures of artificial diets and microalgae or single microalgae. Results indicated that the artificial diet (S2) can serve as substitutes of microalgal diets for P. martensii.     

Growth performance and immune responses of gibel carp, <Emphasis Type="Italic">Carassius auratus gibelio</Emphasis>, fed with graded level of rare earth-chitosan chelate

A 12-week feeding trail was conducted to assess the effect of rare earth-chitosan chelate (RECC) on growth performance and immune responses of gibel carp, Carassius auratus gibelio. Isonitrogenous and isolipid experimental diets were supplemented with graded levels of rare earth-chitosan chelate (RECC 0, 0.8, 4 and 8 g Kg^?1). A total of 720 gibel carps (initial body weight about 14.32 g) were divided randomly into four groups with six replicates, respectively. Fish were fed with the experimental diets three times every day. At the end of the feeding trail, the survival rate was higher than 96 %. Weight gain rate and specific growth rate (SGR) significantly increased with RECC supplementation up to 0.8 g Kg^?1 (P < 0.05) and tended to decline at higher supplementation levels, while feed conversion ratio was not significantly different between groups (P > 0.05). Based on broken-line regression analysis of SGR, the optimum dietary RECC was estimated to be 0.71 g Kg^?1 of the diet. Condition factor and viscerosomatic index were not significantly affected by RECC (P > 0.05), while hepatosomatic index in the group fed with 0.8 g Kg^?1 RECC (3.45 ± 0.10 %) was significantly lower than in other groups (P < 0.05). Plasma ALT was significantly affected by RECC (P < 0.05), while AST was not. Plasma total protein and albumin were increased with RECC supplementation up to 0.8 g Kg^?1 and decreased significantly at higher supplementation levels (P < 0.05). RECC supplementation significantly decreased plasma urea and glucose concentration and increased plasma creatinine concentration significantly (P < 0.05). Respiratory burst activity of phagocytes and myeloperoxidase activity were not significantly different between groups, while superoxide dismutase activity and nitrogen monoxide concentration were increased with the increasing level of RECC in the diets. In conclusion, RECC could enhance growth performance and improve immunity of gibel carp.     

Substitution of soybean meal with fermented <Emphasis Type="Italic">Jatropha</Emphasis> kernel meal: effect on growth performance,body composition,and metabolic enzyme activity of <Emphasis Type="Italic">Labeo rohita</Emphasis>

A 60-day feeding trial was conducted to evaluate the nutritional potential of fermented Jatropha kernel meal (FJKM) in the diets of rohu, Labeo rohita, fingerlings. Four iso-nitrogenous (336 g kg^?1) and iso-energetic (20 MJ kg^?1) diets containing 0, 100, 200, and 300 g kg^?1 FJKM in replacement of 0, 33.3, 66.7, and 100% soybean meal protein, respectively, were prepared and designated as control, T1, T2, and T3, respectively. A total of 180 acclimatized fish (average weight 6.05?±?0.04 g) were equally distributed into four experimental groups in triplicates, and were fed their respective diets to satiation twice daily at 10:00 h and 18:00 h. Higher percent weight gain (%WG); specific growth rate (SGR); protein efficiency ratio (PER); protease, aspartate amino transferase (AST), and alanine amino transferase (ALT) activities; and lower feed conversion ratio (FCR) were recorded in T2 and T3 groups compared to other groups. Whereas hepatosomatic index (HSI), intestinal somatic index (ISI), and amylase, lactate dehydrogenase (LDH), malate dehydrogenase (MDH), superoxide dismutase (SOD), and catalase activities among the various dietary groups did not vary significantly (P?>?0.05). Whole body composition of fish analyzed at the end of the feeding trial exhibited significantly (P?<?0.05) higher ether extract and lower crude protein in the control group compared to the FJKM-fed groups. These results indicated that rohu fingerlings can efficiently utilize FJKM without any detrimental effect on growth performance, nutrient utilization, and metabolic response.     

Physiological response,blood chemistry profile and mucus secretion of red sea bream (<Emphasis Type="Italic">Pagrus major</Emphasis>) fed diets supplemented with <Emphasis Type="Italic">Lactobacillus rhamnosus</Emphasis> under low salinity stress

Environmental stressors caused by inadequate aquaculture management strategies suppress the immune response of fish and make them more susceptible to diseases. Therefore, efforts have been made to relieve stress in fish by using various functional feed additives in the diet, including probiotics. The present work evaluates the effects of Lactobacillus rhamnosus (LR) on physiological stress response, blood chemistry and mucus secretion of red sea bream (Pagrus major) under low salinity stress. Fish were fed four diets supplemented with LR at [0 (LR0), 1 × 10² (LR1), 1 × 10⁴ (LR2) and 1 × 10⁶ (LR3) cells g^?1] for 56 days. Before stress, blood cortisol, urea nitrogen (BUN) and total bilirubin (T-BIL) showed no significant difference (P > 0.05), whereas plasma glucose and triglyceride (TG) of fish-fed LR2 and LR3 diets were significantly lower (P < 0.05) than those of the other groups. Plasma total cholesterol (T-CHO) of fish-fed LR3 diet was significantly (P < 0.05) lower than that of the other groups. Furthermore, total plasma protein, mucus myeloperoxidase activity and the amount of mucus secretion were significantly enhanced in LR-supplemented groups when compared with the control group (P < 0.05). After the application of the low salinity stress test, plasma cortisol, glucose, T-CHO and TG contents in all groups showed an increased trend significantly (P < 0.01) compared to the fish before the stress challenge. However, plasma total protein and the amount of secreted mucus showed a decreased trend in all groups. On the other hand, BUN, T-BIL and mucus myeloperoxidase activity showed no significant difference after exposure to the low salinity stress (P > 0.05). In addition, the fish that received LR-supplemented diets showed significantly higher tolerance against low salinity stress than the fish-fed LR-free diet (P < 0.05). The physiological status and the detected immune responses, including total plasma protein and mucus myeloperoxidase activity in red sea bream, will provide a more comprehensive outlook of the effects of probiotics to relieve stress in fish.     

Comparative nutritional value of <Emphasis Type="Italic">Jatropha curcas</Emphasis> protein isolate and soy protein isolate in common carp

Jatropha seed cake (JSC) is an excellent source of protein but does contain some antinutritional factors (ANF) that can act as toxins and thus negatively affect the growth and health status of fish. While this can limit the use of JSC, detoxified Jatropha protein isolate (DJPI) may be a better option. An 8-week study was performed to evaluate dietary DJPI to common carp Cyprinus carpio. Five iso-nitrogenous diets (crude protein of 38%) were formulated that consisted of a C _ontrol (fish meal (FM) based protein), J ₅₀ or J ₇₅ (50 and 75% of FM protein replaced by DJPI), and S ₅₀ or S ₇₅ (50 and 75% of FM protein replaced by soy protein isolate, SPI) and fed to triplicate groups of 75 carp fingerlings (75; av. wt. ± SD; 11.4 ± 0.25 g). The growth, feeding efficiencies, digestibility, plasma biochemistry, and intestinal enzymes were measured. Results showed that growth performance of fish fed the S ₇₅- or DJPI-based diets were not significantly different from those fed the C _ontrol diet, while carp fed the S ₅₀ had significantly better growth than the J ₇₅ diet. Fish fed the J ₇₅ diet had significantly lower protein and lipid digestibility as well as significantly lower intestinal amylase and protease activities than all other groups. However, all plant protein-based diets led to significantly higher crude protein, crude lipid, and gross energy in the body of common carp compared to the control treatment. Plasma cholesterol and creatinine significantly decreased in the plant protein fed groups, although plasma triglyceride as well as the red blood cells count, hematocrit, albumin, globulin, total plasma protein, and lysozyme activity were higher in plant protein fed groups compared to FM fed group. White blood cells, hemoglobulin concentration, alkaline phosphatase and alanine transaminase activities, and glucose level in blood did not differ significantly among treatments. The results suggest that the DJPI is non-toxic to carp and can be used to replace FM in the diets of common carp up to 75%, but further research to potentially reduce some inherent ANF within this protein source, such as non-starch polysaccharides, may improve nutrient utilization.     

Effect of stocking density on growth performance,serum biochemical parameters,and muscle texture properties of genetically improved farm tilapia, <Emphasis Type="Italic">Oreochromis niloticus</Emphasis>

The objective of this study was to assess the effects of stocking density on growth performance, serum biochemical parameters, and muscle texture properties of genetically improved farmed tilapia (Oreochromis niloticus, GIFT). Juvenile GIFT with an average initial weight of 12.54?±?0.45 g (mean?±?SD) were randomly stocked in 16 tanks (80 L) in a recirculation aquaculture system at four densities of 10 (D1), 20 (D2), 30 (D3), and 40 (D4) fish per tank for 56 days, with quadruplicate for each density. There were no significant differences in water temperature among the four treatments (P?>?0.05). D4 had the significantly lowest dissolved oxygen content (5.52 vs 5.69–6.09 mg L^?1) (P?>?0.05) and pH (6.63 vs 6.87–7.20) (P?<?0.05). NO₂-N and NH₄-N concentrations significantly increased with increasing stocking density (P?<?0.05). Weight gain (WG) and specific growth rates (SGR) decreased with increasing stocking density. The lowest WG (617.20 vs 660.45–747.06%), SGR (3.52 vs 3.62–3.81% day^?1), and highest feed conversion ratio (1.68 vs 1.53–1.58) were observed in D4. Fish at D4 had significantly lower condition factor (3.11 vs 3.29–3.37%) and survival rate (91.25 vs 97.50%) than those from D1 and D2 (P?<?0.05). With increasing stocking density, serum total cholesterol, triglyceride, and total protein concentrations decreased (P <?0.05) and aspartate aminotransferase and alanine aminotransferase activities increased (P <?0.05). D4 fish had higher moisture content (78.80 vs 76.97%) and lower crude protein content (18.14 vs 19.39%) in muscle than D1 fish (P?<?0.05). Compared to D1 and D2, D3 and D4 had lower muscle hardness (1271.54–1294.07 vs 1465.12–1485.65 g), springiness (0.62–0.65 vs 0.70–0.72), gumminess (857.33–885.32 vs 1058.82–1079.28 g), and chewiness (533.04–577.09 vs 757.53–775.69 g) (P <?0.05). High stocking density resulted in growth inhibition, declines in flesh quality, and disturbance to several serum biochemical parameters.     

Complete replacement of fish meal by other animal protein sources on growth performance of <Emphasis Type="Italic">Clarias gariepinus</Emphasis> fingerlings

To completely replace the fish meal by a mixture of earthworm and maggot meals, experimental diets were tested during 42 days on Clarias gariepinus fingerlings. Five isoproteic and isoenergetic diets (40 % crude protein and 17.9 ± 0.3 kJ g^?1) including the control diet (D1) based on fish meal, were formulated. All these diets satisfied the essential amino acids requirements of C. gariepinus fingerlings. These diets were tested on triplicate groups of 50 fishes (initial body weight: 3 ± 0.1 g) bred in tank (0.5 m³). The approximate ratios 2:5; 1:4; 1:12 and 0:1 between the earthworm meal and the maggot meal were used, respectively, to formulate four diets D2, D3, D4 and D5 without fish meal. After the feeding period, significant differences (P < 0.05) were observed on growth, feed utilization between control diet (D1) and test diets (D2–D5). Fish fed earthworm- and maggot-based diets were grown better than those fed the control diet. Survival and feed utilization were not significantly affected by the ratio between earthworm meal and maggot meal in the test diets. Lipid content was higher in carcass and fillet of fishes fed earthworm- and maggot meals-based diets than that of those fed fish meal-based diet. This study indicates that when the ratio 2:5 between the earthworm meal and the maggot meal is used to entirely replace fish meal and the ratio lysine/arginine of the diet is inferior to 1, the growth performances and feed utilization of Clarias gariepinus fingerlings are improved.     

The effect of dietary folic acid on biochemical parameters and gene expression of three heat shock proteins (HSPs) of blunt snout bream (<Emphasis Type="Italic">Megalobrama amblycephala)</Emphasis> fingerling under acute high temperature stress

The effects of dietary folic acid on biochemical parameters and gene expression of three heat shock proteins (HSPs) of blunt snout bream (Megalobrama amblycephala) fingerling under acute high temperature stress. Six dietary folic acid groups (0.0, 0.5, 1.0, 2.0, 5.0, and 10.0) mg/kg diets were designed and assigned into 18 tanks in three replicates each (300 l/tank) and were administered for 10 weeks in a re-circulated water system. The fingerlings with an initial weight of 27.0 ± 0.03 g were fed with their respective diets four times daily. At the end of the experiment, samples were collected before challenge, 0, 24, 72 h, and 7 days. Serum total protein (TP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), cortisol, glucose, complement C3 (C3), complement C4 (C4, immunoglobulin M (IgM) hepatic superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), and the expression of heat shock protein 60 (HSP60), 70 (HSP70), and 90 (HSP90) were studied. The results showed that fish fed with dietary folic acid between 1.0, 2.0, and 5.0 mg/kg significantly (P < 0.05) increased serum TP, C3, C4 hepatic SOD, CAT, and the expression of HSP60, HSP70, and HSP90 before and after temperature challenge of 32 °C. Also, serum ALP, cortisol, glucose, and hepatic MDA were significantly (P < 0.05) reduced by supplementation of dietary folic acid level 1.0, 2.0, and 5.0 mg/kg before and after the same temperature challenge of 32 °C. Before stress, 0, 24, 72 h, and 7 days significantly (P < 0.05) affects serum biochemical parameters, immune and antioxidant capacities, and expression level of three HSPs. Furthermore, there was no statistical evidence to show that dietary folic acid inclusion level and temperature duration have significant interactive effect on serum biochemical parameters, antioxidant parameters, and gene expression level (P > 0.05) of the three HSPs. However, there were statistical significant interactive effect between dietary folic acid inclusion level and temperature duration on serum C3 and C4 (P < 0.05) except IgM (P > 0.05). The present results indicate that supplementation of basal diet from 1.0 mg/kg; 2.0 and 5.0 mg/kg can enhance acute high temperature resistance ability in M. amblycephala fingerling to some degree and improve physiological response, immune and antioxidant capacities, and expression level of three HSPs.     

Effects of the supplementation of vitamin D<Subscript>3</Subscript> on the growth and vitamin D metabolites in juvenile Siberian sturgeon (<Emphasis Type="Italic">Acipenser baerii</Emphasis>)

The objective of this study was to evaluate the effects of the supplementation of vitamin D₃ on the growth, vitamin D metabolites, and osteocalcin secretion in juvenile Siberian sturgeon (Acipenser baerii). A 90-day growth trial was conducted with juvenile Siberian sturgeon (initial body weight 3.47 ± 0.14 g) fed seven isonitrogenous and isoenergetic practical diets (45% CP and 13% lipid) containing 60 (basal diet), 240, 450, 880, 1670, 3300, or 1.0 × 10⁵ IU/kg feed (D60~D 1.0 × 10⁵) vitamin D₃. The results showed that weight gain and specific growth rate increased as the dietary vitamin D₃ levels increased from 450 to 3300 IU/kg (P < 0.05). The fish fed with D1670 and D3300 diets had higher crude lipid and ash levels than the fish fed the D60 diet (P < 0.05). The fish fed D880, D1670, or D3300 diets had higher 25-OH-D₃ and 1,25-(OH)₂-D₃ levels than the fish fed the D60 diet (P < 0.05). The fish fed D880, D1670, D3300, or D1.0 × 10⁵ diets had higher osteocalcin levels than the fish fed the D60 diet (P < 0.05). Based on the broken line method analysis of weight gain and osteocalcin, the dietary vitamin D₃ requirement of juvenile Siberian sturgeon was estimated to be 1683.30 and 1403.27 IU/kg per diet, respectively.     

The effects of starvation on fast-start escape and constant acceleration swimming performance in rose bitterling (<Emphasis Type="Italic">Rhodeus ocellatus</Emphasis>) at two acclimation temperatures

To investigate the effects of starvation and acclimation temperature on the escape ability of juvenile rose bitterling (Rhodeus ocellatus), we measured the fast-start escape and constant acceleration swimming performance of fish fasted for 0 (control), 1 and 2 weeks and half-lethal periods (6 or 4 weeks) at two temperatures (15 and 25 °C). Fish acclimated at a high temperature exhibited shorter response latency (R), higher maximum linear velocity (V _max) and longer escape distance during escape movement (D _120ms) than those at the low temperature. Starvation resulted in a significant decrease in V _max and D _120ms at either low or high temperature and a significant increase in R at only the high temperature in the half-lethal period groups (P < 0.05). The relationship between V _max (Y, m s^?1) and starvation time (X, week) was Y ₁₅ = ?0.062X + 1.568 (r = ?0.665, n = 36, P < 0.001) at low temperature and Y ₂₅ = ?0.091X + 1.755 (r = ?0.391, n = 40, P = 0.013) at high temperature. The relationship between U _cat (Y, cm s^?1) and starvation time (X, week) was Y ₁₅ = ?1.649X + 55.418 (r = ?0.398, n = 34, P = 0.020) at low temperature and Y ₂₅ = ?4.917X + 62.916 (r = ?0.793, n = 33, P < 0.001) at high temperature. The slopes of equations showed a significant difference between low and high temperature (F _1,63 = 9.688, P = 0.003), which may be due to the different energy substrate utilization when faced with food deprivation at different temperatures.     

Evaluation of different <Emphasis Type="Italic">Pyropia yezoensis</Emphasis> extracts as feed additives for growth and immunity of Japanese flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis>

We evaluated the effects of diets incorporating the red algae Pyropia yezoensis, prepared by several different extraction methods, on the growth of juvenile Japanese flounder Paralichthys olivaceus. We assessed growth performance, as well as the levels of amino acids, fatty acids, insulin-like growth factor I (IGF-I) and interleukins (ILs). Four experimental diets were developed based on different methods of processing P. yezoensis. A commercial feed, laver powder (P), high-pressure heat extraction of laver (HPHE) and acid hydrolysis extraction of laver were used as the experimental diets. Three experimental replicates were established for each diet (40 fish/group, body weight 123.7 ± 1.1 g), and the fish were fed for 6 weeks. We found no significant differences in weight gain, specific growth rate or feeding efficiency among the groups (P > 0.05); however, the fish fed HPHE had the greatest growth performance. Fish fed the laver extracts exhibited the highest protein efficiency ratio compared with the control and P groups. The experimental groups fed P. yezoensis extracts had significantly higher levels of IGF-I (P < 0.05) than those of the control group. High levels of IL-2 were found in the P and HPHE groups, IL-12 in the HPHE group, and IL-6 in all experimental groups. Therefore, these results suggest that a P. yezoensis extract improved the growth performance and immunity of Japanese flounder. In particular, the high-pressure heating process was a useful extraction method for preparing a P. yezoensis extract, which had beneficial effects as a dietary supplement in Japanese flounder.     

The effects of starvation and re-feeding on growth and swimming performance of juvenile black carp (<Emphasis Type="Italic">Mylopharyngodon piceus</Emphasis>)

We investigated the effects of starvation and re-feeding on growth and swimming performance and their relationship in juvenile black carp (Mylopharyngodon piceus). We measured the specific growth rate (SGR), resting metabolic rate (RMR) and constant acceleration test speed (U _CAT, the maximum swimming speed at exhaustion by constant acceleration test with 0.1667 cm s^?2 rate) in a treatment group (21 days of starvation then 21 days of re-feeding) and control group (routine feeding) (n = 20). Starvation resulted in a 17 % decrease in body mass of black carp (P < 0.05). After 21 days of re-feeding, body mass was greater than that of pre-starvation but still less than that of the control group at 42 days. During the re-feeding phase, the SGR of the treatment group was higher than that of the control group (P < 0.05). Starvation resulted in a significant decrease in the RMR and U _CAT. After 21 days of re-feeding, both the RMR and U _CAT recovered to the pre-starvation levels. In the control group, individual juvenile black carp displayed strong repeatability of the RMR and U _CAT across the measurement periods (P ≤ 0.002). In the treatment group, RMR showed significant repeatability between pre-starvation and re-feeding (P = 0.007), but not between pre-starvation and starvation or between starvation and re-feeding. U _CAT showed significant repeatability between pre-starvation and starvation (P = 0.006) and between pre-starvation and re-feeding (P = 0.001), but not between starvation and re-feeding. No correlation or only a weak correlation was found between any two variables of RMR, U _CAT and SGR, whereas the increment of the U _CAT (ΔU _CAT) was negatively correlated with that of SGR during the starvation phase (r = ?0.581, n = 20, P = 0.007) and re-feeding phase (r = ?0.568, n = 20, P = 0.009). This suggested that within individual black carp, there is a trade-off between growth and maintenance (or development) of swimming performance under food-limited conditions.     

Role of cyclooxygenase-mediated metabolites in lipid metabolism and expression of some immune-related genes in juvenile grass carp (<Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis>) fed arachidonic acid

Cyclooxygenase (COX) catalyzes the conversion of arachidonic acid (ARA) to prostaglandins, and COX-mediated metabolites play important roles in the regulation of lipid metabolism and immunity in mammals. However, such roles of COX in fish remain largely unknown. In this study, we designed three semi-purified diets, namely ARA-free (control), ARA, and ARA + acetylsalicylic acid (ASA; a COX inhibitor), and used them to feed grass carp (27.65 ± 3.05 g) for 8 weeks. The results showed that dietary ARA significantly increased the amount of ARA in the hepatopancreas, muscle, and kidney (P < 0.05), whereas this increase was reduced by dietary ASA. The hepatopancreatic prostaglandin E₂ content increased in the ARA group, and this increase was inhibited by ASA (P < 0.05). ARA decreased the lipid content in the hepatopancreas, whereas ASA recovered lipid content to a significant level (P < 0.05). ARA significantly decreased the messenger RNA (mRNA) expression levels of fatty acid synthase and stearoyl-CoA desaturase in the hepatopancreas (P < 0.05). However, ASA did not rescue the mRNA expression of these genes (P > 0.05). Interestingly, ARA significantly enhanced the level of peroxisome proliferator-activated receptor α gene expression, and this increase was attenuated by ASA (P < 0.05). Finally, ARA significantly enhanced the mRNA expression of myeloid differentiation factor 88 (MyD88) in the kidney, and ASA attenuated the expression of toll-like receptor 22 and MyD88 (P < 0.05). In conclusion, our findings suggest that COX metabolites play important roles in the inhibition of lipid accumulation in the hepatopancreas of grass carp fed with ARA and that regulation of gene expression promotes lipid catabolism rather than lipogenic activities. Additionally, these eicosanoids might participate in the upregulation of immunity-related genes in the kidney.     

Development of methods for detection of SNPs in <Emphasis Type="Italic">Activin type IIB receptor</Emphasis> and a preliminary study on association between its polymorphism and growth parameters of the Asian seabass <Emphasis Type="Italic">Lates calcarifer</Emphasis>

Molecular markers that allow selection of juveniles and broodstock with a high growth performance are useful for the aquaculture industry. In this study, association between single nucleotide polymorphisms (SNPs) of the Activin type IIB receptor (LcActRIIB) and growth-related parameters of Asian seabass Lates calcarifer juveniles were examined by PCR-direct sequencing. Three SNPs (G>A_c.965+g.7, C>A>T _c.965+g.33 and G>A _c.965+g.189) generating seven composite SNP genotypes (diplotypes) were found in the LcActRIIB gene segment (398 bp). Among three identified SNPs, only a G>A_c.965+g.7 SNP showed significant association with growth traits (average body weight, total length, hepatic weight and hepatosomatic index) of examined fish (P < 0.01). Significant association between diplotypes and growth-related parameters were found. Juveniles exhibiting diplotypes I–V showed greater growth-related parameters than those exhibiting diplotypes VI (P < 0.001; N = 57). Simple methods for rapid genotyping of a G>A_c.965+g.7 SNP were successfully developed based on gel-based bi-allelic PCR amplification of specific alleles (Bi-PASA) and real-time PCR-based PASA analyses (N = 99). Identical genotypes were consistently obtained from different detection methods.     

Growth,hematological and biochemical indices of common carp <Emphasis Type="Italic">Cyprinus carpio</Emphasis> fed diets containing corn gluten meal

The effects of replacing fish meal (FM) with corn gluten meal (CGM) on growth and physiological performance were evaluated in common carp (Cyprinus carpio). Six experimental diets were formulated by substituting 0 (control), 20, 40, 60, 80, and 100% CGM protein for FM protein. The concentrations of dietary protein in the experimental diets were 27.8–29% and the P:E ratios were 14.7–15.46 mg/kJ. Eighteen fish with an initial weight of 13.5?±?0.1 g were allocated to each of 18 circular tanks (450 L) to give triplicate groups for each dietary treatment. The fish were fed to satiation for 8 weeks. At the end of the feeding trial, growth indices, body proximate composition, and hematological and biochemical parameters were measured. Blood samples were taken from six fish in each tank. Final weight and total length were significantly higher in fish fed 100% CGM (27.8?±?1.2 g and 11.9?±?0.3 cm) than for those fed the control (22.7?±?1.4 g and 10.9?±?0.5 cm) or 20% CGM (22.3?±?1.2 g and 11?±?0.4 cm) diets. No effect of FM replacement by CGM was observed for condition factor or hepatosomatic index (P?>?0.05). The highest value of protein productive value (14.31?±?0.65) was observed in fish fed 20% CGM (P?<?0.05). There were no significant differences in percentage body moisture and fat, but percentages of protein and ash were significantly different among experimental groups; the highest values of protein (15.6?±?0.24%) and ash (3.01?±?0.26%) were recorded in fish fed 40% CGM. For hematological parameters, the highest number of white blood cells (4.1?±?0.1?×?10³ mm^?3) was observed in fish fed 100% CGM (P?<?0.05). In addition, the highest hematocrit (42.1?±?0.7%) and triglyceride (294.11?±?23.82 mg dl^?1) were seen in fish fed the diet containing 40% CGM, while 80% CGM gave the highest cholesterol level (204.44?±?9.0 mg dl^?1; P?<?0.05). Replacement of FM with CGM had no negative effects on growth and physiological parameters of common carp fingerlings in this short (8 weeks) trial, suggesting that it may be feasible to replace FM with CGM in diets formulated for juvenile common carp.     

Corn gluten meal in pacu <Emphasis Type="Italic">Piaractus mesopotamicus</Emphasis> diets: effects on growth,haematology, and meat quality

Corn gluten meal (CGM) has a high protein content and absence of antinutrients. However, it has high levels of carotenoids that can cause a yellowing of fish fillets and impair further commercialisation. Graded levels of CGM were incorporated in pacu diets to replace soybean meal (SM) protein. The experimental diets were formulated to be isonitrogenous (220 g kg^?1 digestible protein) and isoenergetic (13.4 MJ kg^?1 digestible energy) with 0, 25, 50, 75, and 100 % replacement of SM protein by CGM protein over a 60-day period. One hundred fish (32.79 ± 3.43 g) were randomly distributed in 20 experimental cages (70 L) placed within five (1000 L) aquaria with continuous water renewal. The experimental design was completely randomised with five treatments and four replicates. A quadratic effect (P < 0.05) was observed for fillet and carcass yields, feed conversion ratio, weight gain, and specific growth rate, with the optimum values for replacement of SM protein by CGM protein estimated as 21.95, 29.13, 30.94, 37.71, and 38.75 %, respectively. There were no differences (P > 0.05) for blood parameters, proximal composition, water-holding capacity, pH, lipid oxidation, and a* value of fillets. The L* and b* values of fillets showed differences (P < 0.05). Replacement between 21.95 and 38.75 % of SM protein by CGM protein (5.37 and 9.48 % of inclusion in the diet) improves the growth performance and body yield of pacu juveniles without altering blood parameters or affecting the fillet quality.     

Effects of starvation and feeding on blood chemistry,fatty acid composition and expression of vitellogenin and fatty acid-binding protein genes in female swimming crab <Emphasis Type="Italic">Portunus trituberculatus</Emphasis> broodstock

Portunus trituberculatus broodstock were stocked in plastic tanks to evaluate the effects of starvation and feeding on gonadal development, blood chemistry, fatty acid composition, and expression of vitellogenin (Vtg) and fatty acid-binding protein genes (FABP) in females. Two treatments (starved and fed) were randomly assigned to triplicate groups of 90 swimming crab broodstock (approximately 230 ± 45 g). In the starved treatment, crabs were starved for 30 days, whereas in the fed treatment crabs were fed once a day with clams. The gonadosomatic index decreased significantly in starved crabs (P < 0.05), as did the serum glucose and cholesterol concentrations; conversely, the total protein concentration in serum significantly increased (P < 0.05). In the ovary, there was a significant relative decline of 18:0, 16:1n-7 and 20:1n-9 fatty acids and relative increases of 20:4n-6, 22:6n-3, 18:1n-9 and 20:5n-3 in starved crabs compared to fed crabs (P < 0.05). Relative expression of Vtg in the ovary decreased significantly in starved crabs (P < 0.05), while there was no significant difference in hepatopancreas Vtg expression between starved and fed crabs (P > 0.05). Starvation suppressed gonadal development in female swimming crab broodstock.     

Molecular mobility of fish flesh measured by low-field nuclear magnetic resonance (LF-NMR) relaxation: effects of freeze–thaw cycles

In this study, the molecular mobility of fish flesh was measured by low field nuclear magnetic resonance (LF-NMR) relaxation. Sardine, tuna and mackerel were frozen at ?40 °C and stored for 1 day (24 h); and then these samples were thawed at room temperature (20 °C). The relaxation of water protons in fish flesh was measured for fresh (i.e., before freezing) and multi-cycle freeze–thaw samples (i.e., up to 12 times). Three domains from different pools of protons (i.e., low-mobile, medium-mobile and high-mobile) were identified from the relaxation curve. The T _2b (low-mobile), T ₂₁ (medium-mobile) and T ₂₂ (high-mobile) indicated the proton populations in the protein molecules, strongly bound water molecules, and weakly bound water molecules, respectively. In all cases, the relaxation time (T _2b: sardine r = 0.736 and p < 0.01, tuna r = 0.857 and p < 0.001, mackerel r = 0.904 and p < 0.001; and T ₂₂: sardine r = 0.956 and p < 0.0001, tuna r = 0.927 and p < 0.0001, mackerel r = 0.890 and p < 0.0001) increased with the freeze–thaw cycles and it reached a nearly constant value after 6 freeze–thaw cycles. The increased relaxation time (i.e., higher mobility) up to 6 freeze–thaw cycles could be due to the increase in proton mobility. However, relaxation time (T ₂₁: sardine r = ?0.510 and p > 0.05, tuna r = 0.162 and p > 0.5, mackerel r = 0.513 and p > 0.01) showed insignificant change with the increase of freeze–thaw cycles, which indicated minimal change in the medium-mobile protons. The results in this study revealed that the changes in proton mobility in the fish flesh during freeze–thaw cycles could be identified using T _2b and T ₂₂ relaxation of LF-NMR.     

Growth performance,feed utilisation and biological indices of Tra catfish (<Emphasis Type="Italic">Pangasianodon hypophthalmus</Emphasis>) cultured in net cages in pond fed diets based on locally available feed resources

The feeding experiment was conducted to evaluate the efficacy of locally available feed resources for Tra catfish (Pangasianodon hypophthalmus) cultured a series of 21 hapa net cages installed in the earthen pond during a 4-month period. The reference diet contained fish meal as the main crude protein (CP) source, whilst in the seven test diets 20–100 % of the fish meal CP was replaced with CP from local feed ingredients: groundnut cake (GNC), cassava leaf meal (CSLM), sweet potato leaf meal (SPLM), soybean meal (SBM), golden apple snail meal (GASM) and shrimp head meal. There were differences (p < 0.05) among diets in final body weight, total weight gain, daily weight gain, specific growth rate, viscera-somatic weight (VSI%), hepato-somatic index (HIS%) and intra-peritoneal fat (IPF%). The respective values for the shrimp head meal diet were numerically highest, followed in descending order by the reference, GASM, GNC, SPLM, CSLM and SBM diet. Food conversion ratio, fish survival rate, total feed intake, feed utilisation, fish fillet and kidney proportions did not differ among the reference and test diets (p > 0.05). However, the viscera-somatic, hepato-somatic, kidney and intra-peritoneal fat indices differed among treatments (p < 0.05). These data show that fish meal protein can be replaced with protein from locally available plant and animal feed ingredient resources in feed cultured in net cages in pond for Tra catfish fingerlings in effectively compromising growth performance, feed utilisation or carcass traits of fish.