Molecular diagnosis of Myxobolus spirosulcatus associated with encephalomyelitis of cultured yellowtail, Seriola quinqueradiata Temminck & Schlegel

Mass mortality of cultured yellowtail, Seriola quinqueradiata, has recently been reported from fish farms in western Japan. Previous studies revealed that diseased fish were characterized by encephalomyelitis and presporogonic stages of a myxosporean‐like parasite in the spinal cord. However, the parasite has remained unidentified because of the lack of mature stages being present. Thus, in the present study, analysis of the small subunit ribosomal DNA (18S rDNA) of the parasite as well as in situ hybridization (ISH) studies using histological sections of the infected tissue was conducted. The 18S rDNA of the myxosporean had higher sequence similarities with those of bile‐duct‐infecting myxosporeans rather than those infecting nervous tissues and was identified as Myxobolus spirosulcatus. The ISH using specific probes demonstrated that the DNA amplified was derived from the multinuclear organisms found in histological sections. A highly sensitive and specific PCR‐based assay for M. spirosulcatus was developed, which revealed a high prevalence of infection in cultured yellowtail that exhibited the clinical signs of encephalomyelitis.     

Effects of dietary supplementation with feeding stimulants on yellowtail Seriola quinqueradiata (Temminck & Schlegel; Carangidae) protein digestion at low water temperatures

Feeding trials were carried out to examine substances that could be used as supplements in low‐protein/high‐lipid diets to improve protein digestion of yellowtails (Seriola quinqueradiata) during winter. Dietary supplementation with synthetic or natural (krill and squid extracts) feeding stimulants (FS) improved feed intake and growth performance of yellowtails. Moreover, the apparent protein digestibility (APD) and pepsin, trypsin and chymotrypsin secretions in fish fed diets supplemented with the different FS were superior to those in fish fed the control diet. These findings suggest that the dietary inclusion of either synthetic or natural FS is a potential tool for improving protein digestion at low water temperatures, and may provide a promising solution for the winter growth retardation experienced in yellowtail culture.     

Seasonal changes in proteolytic enzymes of yellowtail Seriola quinqueradiata (Temminck & Schlegel; Carangidae) fed extruded diets containing different protein and energy levels

Seasonal variations in pepsin, trypsin and chymotrypsin activities and protein digestibility were studied in yellowtails (Seriola quinqueradiata) reared for 1 year with extruded diets containing different levels of protein. Trypsin and chymotrypsin storage levels in the digestive tissues of starved fish were affected by seasonal changes in water temperature. Actual digestion activities of trypsin and chymotrypsin were low at lower water temperatures, but pepsin activity in the stomach tissue was not affected by low temperatures. On the other hand, trypsin and chymotrypsin activities in the intestinal contents were higher during lower water temperature months, while pepsin activity in the stomach contents was low at lower water temperatures. Apparent protein digestibility (APD) did not differ among the dietary treatments in the higher water temperature months, while in colder months it was higher in fish fed diet 1 than in fish fed diets 2 and 3. The APD values reflected pepsin activity in the stomach contents in all sampling months. Therefore, lower APD in colder months seems to be attributed to lower protease activity in the gastric digesta, implying that enhanced pepsin secretion from the stomach tissue might improve protein digestibility and growth performance in yellowtails during winter.     

Effect of dietary taurine supplementation on growth performance of yellowtail juveniles <Emphasis Type="Italic">Seriola quinqueradiata</Emphasis>

ABSTRACT:   The effect of taurine on growth of yellowtail juveniles Seriola quinqueradiata was investigated by a feeding experiment of diets containing various taurine levels. Test diets supplemented with 0, 0.5, 1.0, 1.5 and 2.0% of taurine were prepared. These diets were fed to yellowtail juveniles with an initial mean body weight of 0.5 g for 6 weeks. Supplementation of taurine in the diet of yellowtail improved their growth performance significantly ( P  < 0.05) over the initial 3-week period. The fish fed with the taurine-supplemented diet improved in percent gain and feed efficiency over both 3 and 6 weeks. Taurine content in the muscle proportionally increased with the dietary taurine level. The fish fed without supplemented taurine diet showed higher contents of serine in the muscle. With each increase in the inclusion level of taurine content in the diet, the concentration of serine in the muscle decreased. The cystathionine content in the muscle of each group was unchanged. These results suggest that taurine supplementation in the diet not only improves growth but also affects the sulfur amino acid metabolism of yellowtail juveniles.     

Dietary branched‐chain amino acid supplementation affects growth and hepatic insulin‐like growth factor gene expression in yellowtail,Seriola quinqueradiata

We investigated the effect of dietary supplementation with a branched‐chain amino acid (BCAA) mixture (valine, leucine and isoleucine) on juvenile yellowtail (Seriola quinqueradiata) growth and hepatic insulin‐like growth factor (IGF) gene expression. Total dietary BCAA content was 8.1, 8.5, 9.0 and 9.8 g   kg^?1 for the control, 0.4%, 1.0% and 2.0% BCAA experimental groups, respectively. Body weight was measured on day 77, after which fish were fasted for 3 days and refed; the livers were then removed 3 or 9 h later for analysis of hepatic gene expression. Body weight significantly increased in the BCAA‐supplemented groups. Hepatic expression of IGF1 and IGF2 significantly increased in the 2% BCAA group at 3 and 9 h after refeeding. In the 1% BCAA group, hepatic IGF1 expression tended to be higher at 9 h than in the control group after refeeding. Also, hepatic IGF2 significantly increased at only 9 h after refeeding in the 1% BCAA group. In conclusion, dietary supplementation with crystalline BCAAs increased growth and hepatic expression of IGF1 and IGF2. These results suggest that dietary crystalline BCAA supplementation would be a valuable addition to yellowtail aquaculture practices.     

Effectiveness of l‐ascorbyl‐2‐polyphosphate as an ascorbic acid source for sea cucumber,Apostichopus japonicus

A feeding trial was conducted to determine the adequate dietary ascorbic acid (AsA) levels and the effects on growth, meat quality and antioxidant status of sea cucumber (10.04 ± 0.06 g), Apostichopus japonicus. l ‐ascorbyl‐2‐polyphosphate (35% AsA equivalent) was supplemented separately to the basal diet to obtain five AsA levels, 0, 598, 1473, 4676 and 14340 mg kg^?1 diet respectively. After 60‐day feeding trial, the sea cucumbers fed diets containing 598 and 1473 mg AsA kg^?1 showed significantly higher (P < 0.05) body weight gain and specific growth rate values than the sea cucumbers fed control diets. The sea cucumbers fed diets containing 1473 and 4676 mg AsA kg^?1 showed significantly higher (P < 0.05) hydroxyproline contents than those of the sea cucumbers fed diets containing 0 and 598 mg AsA kg^?1. Antioxidant enzymes such as total antioxidant capacity, superoxide dismutase and glutathione peroxidase showed increasing trends with the increasing dietary AsA levels, but no significant differences (P > 0.05) were observed when the sea cucumbers fed diets with high dietary AsA levels. The content of malondialdehyde had the opposite trend of antioxidant enzymes. In conclusion, the adequate dietary AsA level focusing on growth performance of sea cucumber is between 598 and 1473 mg kg^?1 diet. Furthermore, high level of dietary AsA (between 598 and 4676 mg kg^?1 diet) improved meat quality and antioxidant status.     

Development of a practical soy‐based diet for California yellowtail,Seriola lalandi

Two feeding trials were conducted to initiate the development of a practical soy‐based diet for California yellowtail (YT), Seriola lalandi. The first trial evaluated fish meal (FM), FM + solvent‐extracted soybean meal (SBM) or FM + soy protein concentrate (SPC)‐based diets and a commercial reference diet (Skretting Marine Grower). Final weights (31.8–67.6 g), per cent gain (492.8–1059.9%) and feed conversion ratio (1.11–1.59) all followed a similar response in that fish offered the commercial diet performed significantly better than fish maintained on the other diets. The second trial was designed to evaluate the replacement of FM with increasing levels of soy protein. The basal diet contained 400 g kg^?1 FM and 240 g kg^?1 SBM. The FM was then reduced to 300 g kg^?1, 200 g kg^?1 and 150 g kg^?1 of the diet using SPC as the replacement protein. Final weight (41.2–64.1 g) and per cent gain (110.5–226.5%) followed similar trends with decreases in performance as the FM level was reduced. No gross signs of enteritis were noted, indicating that reduced performance was likely due to nutrient deficiencies or palatability problems rather than an allergic response. Results demonstrate that there is potential to develop reduced FM diets for this species using soy protein.     

The role of dietary L‐ascorbyl‐2‐polyphosphate on the growth and physiological functions of beluga,Huso huso (Linnaeus, 1758)

Effects of dietary ascorbic acid (AA) supplementation were carried out on growth performance and physiological indices in juvenile beluga Huso huso fed different levels of dietary L‐ascorbyl‐2‐polyphosphate as AA source consisting of 0 (control), 100, 200, 400, 800 and 1600 mg AA kg^?1 diet for 16 weeks. No significant differences were found in growth performances between treated groups except for condition factor. Using AA caused significant differences in haemoglobin, number of red blood cells, white blood cells, aspartate aminotranspherase and lactate dehydrogenase, while no significant difference was found in haematocrit, mean corpuscular haemoglobin concentration, differential white blood cells count and alkaline phosphatase activity at the end of the experiment. Lysozyme activity after 8 weeks was significantly affected by dietary treatments with the lowest value in fish fed control diet, but not at completion of 16‐week feeding trial. Liver AA concentration showed a significant increase in correlation with dietary AA increase. Air exposure stress test showed significant differences among the treatments in case of cortisol and glucose concentrations. Also between pre‐ and post stress, significant changes were observed in some of treatments. The lowest and highest levels of cortisol were 9.2 ± 3 (in 200 mg kg^?1) and 16.8 ± 5.1 ng mL^?1 (in control), 6 h post stress (P < 0.05) respectively. Also, the lowest and highest levels of glucose were measured in fish fed 200 and 800 mg AA kg^?1 respectively. The obtained results showed that most growth parameters were not affected by dietary AA, but some physiological and immunological parameters of this species were influenced by this vitamin. We suggest that under rearing conditions, AA should be added to the diet especially during early life stages for improving health status and resistance to stressors.     

Supplementation effect(s) of organic acids and/or lipid to plant protein‐based diets on juvenile yellowtail,Seriola quinqueradiata Temminck et Schlegel 1845, growth and,nitrogen and phosphorus excretion

A feeding experiment was conducted to investigate the effect of organic acids and/or lipid supplementation on growth, utilization and environmental loading of nitrogen (N) and phosphorus (P) in juvenile yellowtail fed fishmeal (FM) and plant protein (PP) diets. Six diets as FM (FM‐based), FM+P (FM with inorganic P), FM+L (FM with lipid), PP+CA (PP with citric acid), PP+L+CA (PP with lipid and citric acid) and PP+L+FA (PP with formic acid) were formulated. Yellowtails were fed each of the diets in duplicate groups; once a day, 6 days a week to near satiation at water temperature 19.0–25.0 °C for 16 weeks. Fishmeal with inorganic P gave the best growth while PP+L+FA the lowest. However, growth increased in PP+CA and PP+L+CA. Addition of lipid significantly increased N and P retention resulting in significant reduction in N and P excretion. Citric acid and FA supplementation to PP diets also increased retention of P; hence, its excretion was lowered. Thus, CA, FA and lipid in juvenile yellowtail diets can help to partially replace FM with PP sources and reduce inorganic P use to minimize environmental loading from aquafeeds.     

Evaluation of l-ascorbyl-2-glucose as the source of vitamin C for juvenile Korean rockfish Sebastes schlegeli (Hilgendorf)

This study was conducted to compare l ‐ascorbyl‐2‐glucose (AA2G) with l ‐ascorbyl‐2‐monophosphate‐Na/Ca (AMP‐Na/Ca) for supplying the dietary vitamin C for juvenile Korean rockfish Sebastes schlegeli (Hilgendorf). Fish were fed one of seven semi‐purified diets containing equivalents of 0, 50, 100 and 200 mg ascorbic acid (AA) kg^?1 diet in the form of AA2G or AMP‐Na/Ca for 12 weeks. After 12 weeks of feeding, weight gain, feed efficiency ratio and survival of fish fed the vitamin C‐free diet were significantly lower than those of fish fed the vitamin C‐supplemented diets in either form. The hepatosomatic index, condition factor and survival of fish fed AMP‐Na/Ca₁₀₀, AMP‐Na/Ca₂₀₀, AA2G₁₀₀ and AA2G₂₀₀ diets were significantly higher than fish fed the vitamin C‐free diet. After 9 weeks of feeding, fish fed the vitamin C‐free diet began to show vitamin C deficiency signs such as anorexia and lethargy. At the end of the 12‐week feeding trial, fish fed the vitamin C‐free diet exhibited vitamin C deficiency signs, e.g., anorexia, scoliosis, exophthalmia and fin haemorrhage. Vitamin C retention in the muscle and liver was similar to those of fish fed AA2G‐ or AMP‐Na/Ca‐supplemented diets. In general, there was no significant difference in the muscle and liver vitamin C concen‐tration in fish fed the AA2G and AMP‐Na/Ca diets at the same supplementation levels.     

Ascorbate-2-sulfate as a dietary vitamin C source for atlantic salmon (Salmo salar): 1. Growth,bioactivity, haematology and humoral immune response

The present experiment shows that ascorbate-2-sulfate (AS) is not equivalent to ascorbic acid (AA) as a dietary vitamin C source for Atlantic salmon (Salmo salar). Within reasonable feed supplemental levels AS does not provide the tissues with adequate supplies of vitamin C to secure optimal physiological functions as demonstrated by biochemical and haematological analyses. AS could not be detected in the liver of fish fed either AA or AS, nor in vitamin C — deprived salmon, suggesting that AS is not the natural storage form of vitamin C in this species. There were no significant differences in antibody production against a soluble artificial antigen (NIP₁₁-LPH) in fish fed 500 and 5000 mg AA/Kg dry diet or equivalent amounts of AS during a period of six weeks at a water temperature of 7.2°C.     

Effects of dietary copper nanoparticles and vitamin C supplementations on growth performance,immune response and stress resistance of red sea bream,Pagrus major

A 60‐day feeding trial was conducted to determine the effects of copper nanoparticles (Cu‐NPs) and vitamin C (VC) on red sea bream. Besides the control diet (D1), six diets were supplemented with Cu‐NPs and VC [0/800 (D2), 0/1,000 (D3), 0/1,200 (D4), 2/800 (D5), 2/1,000 (D6) and 2/1,200 (D7) mg Cu‐NPs/VC per kg]. Cu‐NP was a significant factor on final weight (FBW), weight gain (WG) and specific growth rate (SGR), feed intake (FI), feed (FER) and protein efficiency ratios (PER), protein gain (PG) and protein retention (PR), body protein and lipid contents, protease (PA) and bactericidal activities (BA) and tolerance against stress (LT50%) (p < .05). In addition, BA and LT50% were significantly affected by either Cu‐NPs or VC (p < .05). Fish fed Cu‐NPs or/and VC‐supplemented diets showed higher FBW, WG, SGR, PG, PR, FI, PA, LA and BA values when compared with the control group (p < .05). FER, PER and body lipid content were significantly enhanced in D4, D5, D6 and D7 groups; meanwhile, body protein and LT50% were significantly enhanced in D5, D6 and D7 groups when compared with D1 group (p < .05). In conclusion, dietary Cu‐NPs or/and VC improved growth and health of red sea bream.     

Interactive effects of dietary vitamin C and phospholipid in micro‐bound diet for growth,survival, and stress resistance of larval red sea bream,Pagrus major

This study was conducted to examine the effects of dietary ascorbic acid (AsA) and phospholipid (PL) and their interaction on growth, survival, and stress resistance in red sea bream larvae. Twenty‐six days old red sea bream were fed nine micro‐bound diets supplemented three levels of AsA (0, 800 and 1600 mg kg^?1 diet) and PL (0, 20 and 40 g kg^?1 diet) for 15 days. Dietary AsA and PL were both significant factors on survival rates. There was also an interaction between dietary AsA and PL on survival rate (P < 0.05). The larvae fed 800 or 1600 mg kg^?1 AsA with 40 g kg^?1 PL diets showed the highest survival rate, with values similar to those of the live‐food supplemented group. Stress resistance against low salinity exposure significantly increased with increased dietary level of AsA and PL. However, significant interaction of AsA and PL was not detected. The larvae fed 1600 mg kg^?1 AsA with 40 g kg^?1 PL diet showed the highest stress resistance among all diets, but it was not significantly different than that of larvae fed 800 mg kg^?1 AsA with 40 g kg^?1 PL diet. This study clearly demonstrated that combined use of AsA and PL can improve survival of 26–40 days posthatching red sea bream larvae. Moreover, the present study suggested that 800 mg kg^?1 AsA with 40 g kg^?1 PL in diet was needed for producing high quality seedling under the stressful conditions.     

Effect of l‐ascorbyl‐2‐polyphosphate supplementation on growth performance,body composition,antioxidative capacity and salinity stress tolerance of juvenile Pacific white shrimp,Litopenaeus vannamei

An 8‐week feeding trial was conducted to evaluate the effects of ascorbic acid (AsA), in the form of l ‐ascorbyl‐2‐polyphosphate (LAPP) on growth performance, body composition, antioxidative capacity and salinity stress tolerance of juvenile Pacific white shrimp, Litopenaeus vannamei. Five practical diets (46% crude protein and 7.6% lipid) supplemented with graded levels of AsA (14.64, 48.55, 84.98, 308.36 and 639.27 mg kg^?1 diet) were fed to five replicate groups of L. vannamei (mean initial wet weight 0.57 g). No significant differences were found on growth performance among all treatments. However, whole body lipid content significantly decreased with dietary AsA levels increasing. Activities of total antioxidant capacity, glutathione reductase and glutathione peroxidase were significantly affected by dietary AsA levels. Shrimp fed LAPP‐free diet had higher malondialdehyde content than those fed the diets supplemented with LAPP. Dietary AsA levels higher than 308.36 mg kg^?1 diet increased the survival of shrimps after 1, 2 and 3 h of acute salinity change. Broken‐line regression analysis on survival after 3 h of salinity stress and second‐degree polynomial regression analysis on glutathione reductase data indicated that the optimal dietary AsA requirement of L. vannamei was estimated to be 306.39, 319.75 mg kg^?1 diet respectively.     

The minimum dietary requirement of vitamin C in Atlantic salmon (Salmo salar) fry using Ca ascorbate-2-monophosphate as dietary source

The minimum dietary vitamin C requirement for optimal growth and normal development in Atlantic salmon (Salmo salar) fry at the onset of feeding was studied, using Ca ascorbate-2-monophosphate (AP) as dietary source. The requirement was established by means of a feeding study lasting for 23 weeks from the beginning of feeding. The practical diets used were supplemented with AP at levels of 0, 10, 20, 40, 80 and 160 mg ascorbic acid (AA) equivalents/kg. Growth, mortality, hydroxyproline content in skin and backbone, and AA in liver were recorded to evaluate the results. The results suggest that the minimum dietary requirement for optimal growth and normal development is in the range of 10–20 mg AA equivalents/kg dry diet during the period studied.     

First results of spawning and larval rearing of longfin yellowtail Seriola rivoliana as a fast‐growing candidate for European marine finfish aquaculture diversification

The present study describes the adaptation of longfin yellowtail Seriola rivoliana as broodstock and first larval rearing trials under intensive and semi‐intensive conditions. Fifteen sub‐adults were captured in the South coast of Gran Canaria (Canary Islands, Spain) in June 2007. Fish (initial weight 1.76 ± 0.25 kg) reached a weight of 6.0 ± 1.1 kg in July 2010. Once a year, fish were sampled to determine individual growth in weight and size. In addition, the state of sexual maturity was established based on gonadal biopsies. On the basis of repeated hormonal injection (GnRHa, 20 μg/kg), 10 successful spawns were obtained between July and October 2009, with 92.5 ± 5.5% and 72.6 ± 17.2%, fertilization and egg viability respectively. First results of larval rearing under semi‐intensive conditions, showed an average survival at 30 DAH of 2.5% as compared with 0.5% under intensive conditions. The low survivals under the two rearing conditions in addition to their failure to pass a stress test could be attributed to deficiencies in essential fatty acids as could be seen in both eggs and feeds. Morphometric parameters showed no significant difference between the two rearing systems in 230 DAH larvae.     

Effect of combined fish meal : soybean meal ratio,vitamin C and fish oil supplementations in diet on the growth and reproduction of red swamp crayfish,Procambarus clarkii (Crustacea: Decapoda)

Fish meal (FM), vitamin C (VC) and fish oil (FO) play important roles in the growth and reproduction in aquatic animals. To determine the optimum dietary combinations of FM:SM (soybean meal) ratio, VC and FO supplementations for the growth and reproduction of female crayfish Procambarus clarkii, we used the L9 (3³) orthogonal array, which arranged three factors varied at three levels (FM:SM at 1:3, 1:2 and 1:1.5; VC supplementation at 0.02%, 0.04% and 0.06%; FO inclusion at 1.90%, 2.85% and 3.80%). Range analysis showed that the highest weight gain (WG) of the crayfish occurred at FM:SM of 1:1.5, VC supplementation of 0.04% and FO inclusion of 1.90%, and the maximum gonadosomatic index (GSI), egg laying amount (ELA) and spawning rate (SR) were obtained at FM:SM 1:1.5, VC supplementation 0.06% and FO inclusion 3.80%. Analysis of variance indicated that all the three factors had no significant effects on WG. Fish oil supplementation significantly affected GSI, ELA and SR (P<0.05), while FM:SM did not significantly influence any of these indicators. Taking the cost into consideration, we suggest that a dietary FM:SM of 1:3, VC supplementation 0.06% and FO inclusion 3.80% might be the optimal combinations for the reproduction of P. clarkii.     

Effect of dietary lipid source and vitamin E on growth,non‐specific immune response and resistance to Aeromonas hydrophila challenge of Chinese mitten crab Eriocheir sinensis

Six purified diets were formulated to contain three lipid sources, fish oil (FO), linseed oil (LO) and soybean oil (SO), at 6% diet lipid crossing two levels of vitamin E (100 and 300 mg α‐tocopheryl acetate/kg diet) for each lipid source (FO100, FO300, LO100, LO300, SO100, SO300). The juvenile Chinese mitten crab, Eriocheir sinensis, respectively, fed on these diets with four replicates for 6 weeks. The crab weight gain (WG) and specific growth rate (SGR) were significantly affected by dietary lipid sources. No difference was found between the crabs fed two levels of vitamin E, but the WG and SGR were numerically higher in crab fed 300 mg/kg vitamin E than those fed the other level of vitamin E. The lipid source and vitamin E level could affect fatty acid composition in the hepatopancreas. The contents of saturated fatty acids (SAFA) and n‐3HUFA were significantly higher in the crab‐fed fish oil. The highest contents of n‐6PUFA and n‐3PUFA were found in the crab‐fed soybean oil and linseed oil respectively. The contents of SAFA, n‐3HUFA and n‐3PUFA were higher in the 300 mg/kg vitamin E treatment. A lower malondialdehyde (MDA) content and higher phenoloxidase (PO) activity were observed in the crab fed 300 mg/kg vitamin E. The results of this study indicate that the Chinese mitten crab fed the diet with 6% fish oil and 300 mg/kg vitamin E showed better growth, antioxidant capacity and resistance to Aeromonas hydrophila.