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 taurine levels in broodstock diet on reproductive performance of yellowtail Seriola quinqueradiata

The effect of dietary taurine was investigated on reproductive performance in yellowtail Seriola quinqueradiata. Two-year-old fish of average body weight 6.1kg were fed on diets containing three levels of taurine (T-0, T-0.5 or T-1.0%) for 5 months prior to spawning. For spawning investigations, fish were induced to maturity by human chorionic gonadotrop in injection (600 IU/kg-fish) and artificially inseminated. Oocyte growth improved significantly (P<0.05) with the increase of dietary taurine. The collection of eggs from females reared on the T-0% diet was not successful. The success rate of spawning for females fed on T-0.5 and 1.0% diets was one out of six, and six out of seven, respectively. The taurine levels of the liver and serum in the T-0% diet group were much lower than that in the T-0.5 and T-1.0% diet groups (P<0.05). Fish fed the T-0% diet showed higher contents of serine in the liver and serum. The taurine content of the ovary was not significantly different among the different dietary treatments. These results indicate that taurine has a positive effect on the improvement of spawning performance of yellowtail.     

Experimental induction of nocardiosis in yellowtail, Seriola quinqueradiata Temminck and Schlegel by artificial challenge

Challenge methods for inducing nocardiosis, caused by Nocardia seriolae in yellowtail, Seriola quinqueradiata, were evaluated. The first method involved intraperitoneal injection of 1.1 x 10(2), x 10(3) and x 10(4) cfu of N. seriolae; the second was by bath immersion with three different concentrations of bacterial suspension for 10 min; the third was by intradermal injection using a multipuncture device; the fourth was by oral administration using a tube; the fifth was based on the co-habitation of uninfected fish with others that had been artificially infected, i.e. intraperitoneally or by immersion for 10 min. The LD(50) values for the infection trials were 1.9 x 10(2) (intraperitoneal injection), 1.5 x 10(4) mL(-1) (immersion), 4.3 x 10(6) (intradermal injection) and 1.7 x 10(7) (oral administration). In the co-habitation challenge, mortalities were 70% and 50% in groups of non-infected fish mixed with fish infected by the i.p. injection and immersion methods, respectively. Fish challenged by intraperitoneal injection developed adhesions. Fish challenged by other methods did not show any gross clinical signs and moribund fish were similar to natural infection.     

Effects of dietary phosphorus level on non-faecal phosphorus excretion from yellowtail (Seriola quinqueradiata Temminck & Schlegel) fed purified and practical diets

Non-faecal phosphorus (P) was determined for large yellowtail to estimate a minimum available P requirement (Experiment  1) and to justify inorganic P supplementation in a fish meal-based diet (Experiment 2). In Experiment 1, purified diets with incremental P concentrations were fed to yellowtail (mean weight 917 g) at a feeding rate of 1.5% of body weight. The peaks of non-faecal P excretion appeared 5–6 h after feeding in fish fed more than 4.5 g available P kg⁻¹ dry diet. Broken-line analysis indicated that the minimum available P requirement was 4.4 g kg⁻¹ dry diet. In Experiment 2, a purified diet (PR) containing 6.5 g available P kg⁻¹ and a fish meal-based diet with (F1) and without (F0) additional phosphorus were fed to yellowtail (mean weight 1.1 kg) at 1.5% (PR) and 2% (F0 and F1) feeding rates respectively. There was no significant difference in P excretion between fish fed the F0 (5.5 g soluble P kg⁻¹ dry diet) and the PR diet. However, significantly higher (34.5%) amounts of non-faecal P excretions (7.4 g soluble P kg⁻¹ dry diet) were found in fish fed F1 compared with the F0 diet. This suggested that there was an excess of dietary P in the F1 diet and that supplementation is not needed in fish meal-based diets for large yellowtail.     

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.     

Effects of replacing fish oil with canola oil on the growth performance,fatty acid composition and metabolic enzyme activity of juvenile yellowtail Seriola quinqueradiata (Temminck & Schlegel, 1845)

In this study, fish oil (FO) was replaced with canola oil (CO) in juvenile yellowtail diets to establish the optimal replacement levels that ensure adequate feed quality for aquaculture. Juvenile yellowtails (initial body weight: 104.7 g) were fed one of four diets for 10 weeks: 100% FO (FO group) or FO replacement with 25%, 50% or 100% CO (CO25, CO50 and CO100 groups respectively). Body weight, specific growth rate and feed efficiency were significantly affected by the replacement of FO with CO. The final body weight of the CO100 group was significantly lower than that of the FO (control) and CO50 groups. Furthermore, the CO50 group showed significantly higher feed efficiency than the FO group during the first 4 weeks (long photoperiod and high water temperature conditions). Increasing dietary CO increased serum glucose and triglyceride concentrations in a dose‐dependent manner. Regarding the proximate composition of fillet and liver, only crude protein in fillet was affected by the replacement of FO with CO. Fatty acid compositions of fillet and liver were correlated to the amount of CO in each diet. The activities of the metabolic enzymes phosphofructokinase and alanine aminotransferase were lowest in the CO25 and CO50 groups respectively. Thus, FO replacement with CO affected growth performance, serum components, fatty acid composition and the activity of metabolic enzymes. Overall, results obtained in the present study suggest that partial replacement (50%) of FO with CO is effective and might increase juvenile yellowtail growth under long photoperiod and/or high water temperature conditions.     

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.     

Effects of dietary phospholipid level and fraction on the feed intake of non‐fish meal diet in yellowtail,Seriola quinqueradiata Temminck & Schlegel, 1845

Three feeding studies were conducted to investigate whether dietary soybean lecithin (SBL) enhanced feed intake of non‐fish meal, SPC‐based diet, in yellowtail, and also to clarify which fraction of phospholipid (PL) was effective to enhance feed intake. In experiments 1 and 2, fish were fed SBL level at 0%–5%, and it was revealed that the daily feeding rate (DFR) of fish was significantly higher for SBL 2% than for SBL 0%, while a further increase of SBL in the diet did not additionally increase the DFR. Moreover, the DFR of fish fed a diet without feeding stimulants (FS) was the lowest among the dietary treatments, indicating that the SBL does not have an effect of enhancing feed intake in yellowtail. In experiment 3, fish were fed a diet with several products containing PL at 2%. Phosphatidylcholine and phosphatidylinositol concentrations in these products positively correlated to the DFR of fish. This result revealed that these two fractions in the SBL are effective for enhancing feed intake of yellowtail when FS is present in the diet.     

Effectiveness of l‐ascorbyl‐2‐monophosphate Na/Ca as a vitamin C source for yellowtail Seriola quinqueradiata juveniles

A feeding experiment was conducted to examine the efficacy of a vitamin C (L‐ascorbic acid, AsA) derivative, l ‐ascorbyl‐2‐monophosphate Na/Ca (AMP‐Na/Ca) by comparing to l ‐ascorbyl‐2‐monophosphate‐Mg (AMP‐Mg) in terms of growth, AsA content in tissues, and hematology in yellowtail Seriola quinqueradiata juvenile (1.10 ± 0.01 g). Furthermore, a stress resistance test for the species fed AMP‐Na/Ca was also conducted. In experiment 1, 11 test pellet diets with different levels of AMP‐Na/Ca (0, 12, 43, 88, 440 and 881 mg AsA kg^?1diet) and AMP‐Mg (0, 16, 52, 106, 595 and 1164 mg AsA kg^?1 diet) were formulated and fed to the yellowtail three times per day for 50 days. In both the vitamin C sources, the survival rates of the fish fed diets without supplemental AsA were lower than 50% at day 20, and more than 50% mortality occurred in fish that fed the diets containing 12 or 16 mg AsA kg^?1 after day 30. However, no significant differences were detected in survival and growth among fish that fed the diets containing more than 43 mg AsA kg^?1. Liver and brain AsA concentrations generally increased with increasing dietary AsA level in both sources. In experiment 2, test diets were formulated to contain 43, 88, 440 and 881 mg AsA kg^?1 using AMP‐Na/Ca, and after 60 days of feeding, yellowtail juveniles were subjected to low salinity and air exposure stress. The fish that received diets with 440 mg AsA kg^?1 showed significantly higher tolerance to low salinity stress and higher survival rate in air exposure stress than those of other groups. The present study demonstrated that yellowtail juveniles could utilize AMP‐Na/Ca as an AsA source like AMP‐Mg, and that supplementation of 43–52 mg AsA kg^?1 diet was optimum for normal growth. However, this study showed that dietary inclusion of 440 mg AsA kg^?1 would be necessary to improve stress resistance of this species.     

Pigment deficiency correction in shrimp broodstock using Spirulina as a carotenoid source

In this study, an effort was made to cure the pigment deficiency syndrome (PDS) exhibited by pond‐reared Fenneropenaeus indicus (H.Milne Edwards) broodstock, using the blue‐green algae Spirulina as a carotenoid source in diet. The PDS symptoms included bleaching of ovary and reduced early larval performance. Biochemical analysis indicated reduced quantity of total carotenoids in egg and nauplii from the females with PDS. When PDS females were fed 30 g kg^?1Spirulina‐supplemented diet immediately after the appearance of symptoms, the egg and nauplii carotenoid values improved gradually and attained the pre‐PDS values after a minimum of 4 week period. Spirulina supplementation also improved significantly (P < 0.05) the spawn quality (hatch per spawn, nauplii per spawn, nauplii viability) and larval quality indices (P < 0.05). The egg and nauplii protein values exhibited a trend similar to carotenoids, confirming the influence of latter on protein stability previously reported by researchers. The study confirms the bioavailability of carotenoids from Spirulina to shrimp broodstock and recommends its regular inclusion in diet to preclude carotenoid deficiency‐related problems in shrimp hatcheries.     

The abundance of juvenile yellowtail (Seriola quinqueradiata) near the Kuroshio: the roles of drifting seaweed and regional hydrography

We assess the effect of drifting seaweed (Sargassum sp.) biomass, geography and hydrography on juvenile yellowtail (Seriola quinqueradiata) abundance variation off the southeast coast of Japan, near the Kuroshio Current. The amount of drifting seaweed mats progressively increased northeastward into the cooler, coastal waters. Frontal structure indexed using a station‐to‐station ΔSST did not explain spatial variation in the seaweed mat distribution, although the western extent of the Kuroshio Current appeared to act as a boundary. Juvenile yellowtail constituted 51–62% of the fish collected in association with drifting seaweed mats in April 1996 and 1997 and 29% in June 1996. The abundance of juvenile yellowtail was positively correlated with seaweed biomass. The geographic distribution of juvenile yellowtail associated with drifting mats varied among sampling periods, being more southwesterly in April and more northeasterly in June. Simple multiple regression models based on seaweed biomass and geographic distribution (latitude) explained between 35% and 43% of the variation in juvenile yellowtail abundance in spring. Associations with spatial and temporal variations in hydrographic conditions did not contribute to explained variation in a meaningful manner. The results presented here indicate that, off the southeast coast of Japan during April, yellowtail juveniles are likely to be most abundant when seaweed biomass is high, occur offshore, and are bounded by the western extent of the Kuroshio Current near the 19–20°C SST isotherm.     

Utilization of soybean meal as an alternative protein source in the Mediterranean yellowtail, Seriola dumerili

The availability of defatted soybean meal as a substitute for fish meal was evaluated in juvenile (450 g on average) yellowtail fed diets containing several levels of soybean meal (from 20 to 50%). After 153 days, liveweight averages were 980, 925, 795 and 670 g for fish fed diets in which fish meal was partially substituted with 20, 30, 40 and 50% soybean meal, and feed conversion ratios were 2.79, 3.09, 4.57 and 6.52, respectively. Regression analysis showed a negative effect of inclusion of soybean, although fish fed diets containing 20% and 30% of soybean did not present statistical differences and grew significantly more, and had a better feed conversion ratio, than fish fed 40 or 50% diets. Likewise, muscle protein level was lower and lipid content was higher in fish fed 20 or 30% soybean. No differences were obtained for protein digestibility coefficients of experimental diets.     

Effect of squid meal in dry pellets on the spawning performance of striped jack Pseudocaranx dentex

SUMMARY: This study was carried out to investigate the effects of dietary squid meal or a combination of squid meal and krill meal as part of the protein source in dry pellets on the spawning of striped jack Pseudocaranx dentex . Five months prior to spawning, 7-year-old fish were divided into three groups of 10 fish each (male : female ratio, 5 : 5). The control group was fed a raw fish mix (RF) and the other two groups were fed either steam-dry pellets with squid meal replacing 50% of their fish meal (fs-DP) or steam-dry pellets containing equal proportions of fish, squid and krill meals (fsk-DP). Feeding was carried out once every other day in 5 × 5 × 5 m floating net cages and the fish were transferred to 65 m³ indoor tanks for spawning. Eggs and yolksac larvae produced were evaluated for their quality and those obtained during the first 2 weeks of spawning were sampled for chemical analysis. The fish had an average bodyweight of 3.5 ± 0.4 kg at spawning. Although egg production of the RF group was significantly higher ( P < 0.05) than that of the dry pellet groups, the fs-DP group produced the best quality eggs with higher fertilization and hatching rates. The fsk-DP group had the lowest egg production and quality. Lipid classes and fatty acid compositions of eggs and yolksac larvae were dependent on the broodstock diets. These results show that the combination of fish meal and squid meal in dry pellets for striped jack improved egg quality but not production whereas the combination of fish meal, squid meal and krill meal was not effective.     

Response changes with growth in the gustatory receptors of young yellowtails (Seriola quinqueradiata)

SUMMARY: Laboratory-hatched juvenile yellowtails (mean fork length, 9.3 cm) were reared for 2 months in two groups on different kinds of food, mysid and squid in one group and fish in the other, and the gustatory neural responses to seven chemicals (adenosine-5'-monophosphate, alanine, arginine, betaine, proline, tryptophan, and valine) were recorded in fish before and after the 2-month rearing period. Large variances in the responses to some chemicals were noticed in the juveniles before the rearing experiment. Mann–Whitney U -tests on the neural responses indicated significant changes in the magnitude of the responses to valine and some other chemicals relative to that for proline or tryptophan between the pre- and the two post-rearing groups. No significant differences in the response magnitude for the seven chemicals were observed for the two post-rearing groups suggesting that the response changes during the 2-month rearing might have been intrinsic and not due to specific food items in the diet. The dose–response relationship for some chemicals was also examined in the juveniles before and after the 2-month rearing. A slight lowering of threshold was noticed for alanine and valine after the rearing. Data on the responses of wild yellowtails were in support of the changing responsiveness of gustatory receptors during development of juvenile yellowtails.