Immune response of largemouth bass, <Emphasis Type="Italic">Micropterus salmoides</Emphasis>, to whole cells of different <Emphasis Type="Italic">Nocardia seriolae</Emphasis> strains

Largemouth bass Micropterus salmoides were immunized with four different N. seriolae strains—two α-glucosidase-positive (961113, KU040801) and two α-glucosidase-negative (94260, OTTS) strains—along with Freund’s incomplete adjuvant. After primary immunization (week 0), a booster was administered at weeks 4 and 8. Nonspecific immune responses to multiple immunizations with the different N. seriolae strains were determined based on serum lysozyme activity and nitroblue tetrazolium (NBT)-positive cells in peripheral blood. The serum lysozyme activity and NBT-positive cells in peripheral blood were not significantly increased even after the two booster immunizations. Specific antibody responses against N. seriolae cells were investigated by enzyme-linked immunosorbent assay. At 4 weeks after immunization, all groups immunized with N. seriolae antigens showed significant increases in their specific antibody levels. The sera from fish immunized with different N. seriolae strains exhibited reactivity with N. seriolae sonicated antigens of 28, 30, 36 and 84 kDa by western blot analysis. After two boosters, fish were challenged with live N. seriolae to assess the vaccine’s efficacy; however, multiple injections of the N. seriolae strains did not reduce mortality, irrespective of the bacterin.     

Optimisation of larval culture of the mussel <Emphasis Type="Italic">Mytilus</Emphasis><Emphasis Type="Italic">edulis</Emphasis> (L.)

The blue mussel Mytilus edulis is a commercially important species whose fishery and culture generally relies on natural spat collection. Hatchery-production could provide an alternative source of seed, enabling reliable expansion of the industry. Mussel spawning and larval rearing trials were carried out to optimise elements of hatchery production. Culturing fertilised eggs at low density (20–200 eggs cm⁻²) rather than high density (400–720 eggs cm⁻²) significantly improved the quality of first veliger larvae and differences in this improvement were evident between the eggs from different females (maternal effects). Veliger larval growth at 17 or 21°C was significantly faster than growth at 14°C. Feeding veliger larvae an identical total ration either daily or at 2–3 day intervals did not significantly affect their growth. Different microalgal diets (1: Isochrysis sp. (clone T-ISO), 2: Chaetoceros calcitrans forma pumilus, 3: C. muelleri, 4: mixed Isochrysis sp. (clone T-ISO) and C. calcitrans f. pumilus, and 5: mixed Isochrysis sp. (clone T-ISO) and C. muelleri) were tested on veliger larval growth and mixed diets outperformed single-species diets.     

The impact of net-isolated polyculture of tilapia (<Emphasis Type="Italic">Oreochromis niloticus</Emphasis>) on plankton community in saline–alkaline pond of shrimp (<Emphasis Type="Italic">Penaeus vannamei</Emphasis>)

We used 12 land-based experimental enclosures (6 m × 5 m) in a saline–alkaline pond of shrimp (Penaeus vannamei) to determine the impact of net-isolated polyculture of tilapia (Oreochromis niloticus) on plankton communities for 40 days. Tilapias were stocked in net cages suspended in enclosures, in polyculture ponds including tilapia and shrimp. Four tilapia biomass were tested: 0, 39, 115 and 227 g m⁻². Shrimp stocking biomass were 0.7 g m⁻² in all treatments. There were three replicates in each treatment. Our results showed that the presence of tilapia significantly reduced phytoplankton biomass directly through predation and indirectly through top-down effect. The stocking of tilapia reduced zooplankton biomass, particularly rotifer biomass. However, copepod biomass was not been significantly affected. So, net-isolated polyculture of tilapia can thus have a strong impact on phytoplankton allowing the co-existence of large numbers of copepods with planktivorous fish and improving the water quality of shrimp ponds.     

Use of vitamin C as an immunostimulant. Effect on growth,nutritional quality,and immune response of <Emphasis Type="Italic">Labeo rohita</Emphasis> (Ham.)

Teleost fish lack the enzyme for endogenous synthesis of ascorbic acid (AA), an essential micronutrient for fish. The aim of this study was to examine the effect of higher levels of dietary vitamin C on growth, nutritional quality, and immunomodulation in the Indian major carp, rohu (Labeo rohita). Four groups of L. rohita were fed experimental diets containing either no vitamin C (control) or supplemented with vitamin C at 500 mg kg⁻¹ (Exp-1), 1000 mg kg⁻¹ (Exp-2), or 1500 mg kg⁻¹ (Exp-3) for 60 days. Growth parameters (NWG, ADG, and SGR), serological parameters (TSP, TSA, TSG, and A:G), haematological parameters (TLC, TEC, Hct, MCV, and MCH), and different non-specific immunological parameters (PR, PI, respiratory burst activity, and bactericidal activity) were evaluated during the experimental trial. Fish fed a vitamin C-supplemented diet showed higher specific growth rate (SGR) up to 1000 mg kg⁻¹ compared with control fish. Different haematological and serological parameters along with non-specific immune parameters were influenced by vitamin C supplementation. Among the non-specific immune parameters phagocytic activity (PR and PI) and respiratory burst activity (NBT cells) were significantly (P ≤ 0.05) enhanced by increasing doses of vitamin C supplementation. Higher levels of dietary vitamin C significantly (P ≤ 0.05) enhanced protection against Aeromonas hydrophila (AH1) infection compared with controls. Results from this study help to establish the beneficial effect of vitamin C on growth and immunmodulation in rohu (L. rohita).     

The harvest of the freshwater crayfish <Emphasis Type="Italic">Astacus leptodactylus</Emphasis> Eschscholtz in Turkey: harvest history,impact of crayfish plague,and present distribution of harvested populations

This review focuses on the present distribution of populations of the crayfish Astacus leptodactylus that are harvested in Turkey. It also examines the history of this harvest and the impact that crayfish plague has had on them. Crayfish plague, caused by the fungus-like organism, Aphanomyces astaci Schikora, 1906, is a severe parasite of freshwater crayfish and has caused a lot of damage to A. leptodactylus populations in Turkey since 1984. Turkey was the largest provider of A. leptodactylus to Western Europe from 1970 (or possibly earlier) until 1986. For example, the peak production was reached in the early 1980s, with over 5,000 tonnes being exported in 1984. On the other hand, as a result of the crayfish plague the harvest of A. leptodactylus was reduced severely in most populations in Turkey after 1985. The harvest was only 320 tonnes in 1991. After the occurrence of crayfish plague in Turkey, in order to increase crayfish production uncontrolled A. leptodactylus stockings have been carried out in many waterbodies throughout Turkey. These introductions have caused an increase in the number of A. leptodactylus populations, but exploitation of A. leptodactylus is still under the pressure of the plague, although there has been a steady increase in crayfish production in recent years. The harvest increased to 2,317 tonnes in 2004. Fortunately, among those populations affected by crayfish plague, large amounts of A. leptodactylus can still be harvested from three lakes, ?znik (Bursa), E?irdir (Isparta) and Çivril (Denizli). Thus, it seems that A. leptodactylus has a degree of resistance to crayfish plague. It is therefore interesting to investigate the resistance of A. leptodactylus caught from these populations to crayfish plague.     

Effects of triploidy on the Caspian salmon <Emphasis Type="Italic">Salmo trutta caspius</Emphasis> haematology

The aim of this study was a comparison of key haematological features of diploid (2n) and triploid (3n) Caspian salmon (Salmo trutta caspius). Morphometric indices of erythrocytes were determined on blood smears by light microscopy. Triploidy significantly (P < 0.001) increased all morphometric indices measured in the erythrocytes including cell size, cell surface area, and cell volume. The increase in cell size was larger for the major (27%) axis than for the minor (22%) axis, thus making erythrocytes of 3n Caspian salmon more ellipsoidal. The estimated increase in erythrocyte nuclear volume (87%) was bigger than the theoretical expected 50% increase. Haematological indices were measured manually by hemocytometry. Triploids had lower numbers of red blood cells (RBC: 1,120,000 cells/mL in 2n vs. 700,000 cells/mL in 3n; P < 0.001) but they were larger in size (mean erythrocytic volume [MEV]: 363.1 nm³ in 2n vs. 483.3 nm³ in 3n; P < 0.001). The decrease in RBC number was not compensated by the increase in MEV and, thus, triploidy affected the haematocrit (Hct: 38.8% in 2n vs. 33.06% in 3n; P < 0.05). Total blood hemoglobin concentration was lower in triploid fish (Hb: 9.9 g/dL in 2n vs. 8.9 g/dL in 3n; P < 0.05). In contrast, mean erythrocytic hemoglobin (MEH: 95 μg in 2n vs. 133.2 μg in 3n; P < 0.001) was higher for 3n Caspian salmon as a result of their larger erythrocytes, although MEH concentration (MEHC: 0.26 g/dL in 2n vs. 0.27 g/dL in 3n) did not significantly differ (P > 0.05). White blood cell (WBC) counts (lymphocytes and neutrophiles) were measured and WBC/RBC ratios were calculated. There were no significant differences in WBC (15,710 cells/mL in 2n vs. 12,683 cells/mL in 3n; P > 0.05), lymphocytes, and neutrophils as %WBC as well as WBC/RBC ratios between two ploidy levels (P > 0.05). Triploid Caspian salmon showed higher erythrocyte abnormalities such as ‘twisted’, ‘tailed’, and ‘anucleated’ cells as well as high portions of immature RBC in blood smears in comparison with diploids (P < 0.001).     

Elemental analysis of otoliths of Japanese anchovy: trial to discriminate between Seto Inland Sea and Pacific stock

The ability to discriminate local stocks of Japanese anchovy Engraulis japonicus was assessed based on data from four elements (K, Na, P, and Sr) using an electron probe micro analyzer (EPMA) and data from three elements (Ba, Mn, and Sr) using inductively coupled plasma mass spectrometry (ICP-MS) from the otoliths of 40 anchovy (23.6–47.0 mm body length). Anchovy were caught at three sites (Aki-nada, Hiuchi-nada, and Osaka Bay) in the Seto Inland Sea, and one site (Kuroshio extension) in the Pacific Ocean in 2002. In order to discriminate different spawning grounds, EPMA data from the core portion (from core to 30 μm in the core-posterior axis) were used. Results showed that it was difficult to discriminate between the Seto Inland Sea and the Pacific anchovy by EPMA data. Conversely, it was possible to discriminate between the Seto Inland Sea and the Pacific anchovy by ICP-MS data from bulk otoliths. Our results showed that Mn contents of otoliths using ICP-MS discriminate between spawning grounds most, and Ba and Sr discriminate less. The difference in elemental compositions in anchovy otoliths between the Seto Inland Sea and the Pacific Ocean might be reflected by cumulative experienced elemental composition of ambient sea water during life history between the Seto Inland Sea and the Pacific anchovy.     

Suppression of fish meat softening by strict control of storage temperature

ABSTRACT:   Changes in meat firmness and structure of cultured yellowtail at several storage temperatures, from −1.5°C to 10°C, were compared. Firmness decreased less at −1.5°C and 10°C than at the other temperatures. During storage at temperatures from −0.5 to 4.0°C, expansion of intercellular space, which indicates weakening of the connecting force between muscle cells, began to increase after 8 h storage, and the ratio of space to the whole area increased by 3.4–4.9 times after 24 h of storage. However, at −1.5°C and 10°C, the increase of intercellular space was suppressed, and the ratio was 2.4–2.6 times, even after 24 h storage. Collagen fibrils connecting muscle cells disintegrated after 24 h storage at all storage temperatures. Conversely, in 10°C storage, the fine structure of muscle, except for collagen fibrils, was maintained better than at other storage temperatures. These findings indicate that −1.5 and 10°C storage could maintain fish-meat quality for longer periods than storage at −0.5, 1.0, or 4.0°C, the normal refrigeration temperature.     

Guanine-cytosine contents of the host and symbiont cDNA in a symbiotic coral

ABSTRACT:   Hermatypic (reef-building) corals harbor dinoflagellate endo-symbionts Symbiodinium spp. In studying gene expression in such symbiotic corals, problems arise regarding how to distinguish the coral and symbiont mRNA, and how to estimate their fractions in the mRNA population of the holobiont (symbiotic complex of the coral and Symbiodinium cells). In this study, these issues were addressed using juveniles of hermatypic coral Acropora tenuis in symbiosis with Symbiodinium cells of strain PL-TS-1. First, the guanine-cytosine (GC) contents were determined in expressed sequence tags (EST) from PL-TS-1 cells cultured in vitro and symbiont-free larvae of A. tenuis , and their average GC contents were found to be significantly different. The average GC content of the EST from the holobiont was much closer to that of A. tenuis larvae, suggesting that the majority (>90%) of mRNA isolated from the holobiont originated in the host. In protein-coding sequences, little overlap was observed between the GC-content distributions of PL-TS-1 cells and A. tenuis larvae. All of the coding sequences ( n  = 59) found in the A. tenuis EST had GC contents below 0.5, whereas the GC content exceeded 0.5 in the majority (43/44) of coding sequences from the nuclear genome of PL-TS-1 cells.     

Effects of silvering state on induced maturation and spawning in wild female Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis>

ABSTRACT:   The effects of silvering state of wild female Japanese eels Anguilla japonica on the success of induced maturation and the following spawning were examined. Thirty-eight females, collected in Mikawa Bay, were divided into four stages based on their silvering state: yellow (Y1), late-yellow (Y2), silver (S1) and late silver eels (S2). Despite injections of salmon pituitary extract (SPE) through the standard technique, Y1 and Y2 eels did not respond to the treatment with undeveloped gonad (gonad-somatic index [GSI]: 0.3–0.9), and all these females died by 5 weeks, probably due to an abnormal physiological condition. Most S1 (81%) and S2 eels (100%) matured completely (GSI: 17.8–51.4), and finally spawned successfully (69% for S1, 89% for S2). S2 eels fully matured with oocytes of over 750 μm in diameter by significantly smaller number of injections of SPE (5–6 times) than the case of S1 eels (6–8 times). The amount of eggs released by S2 eels (0.65 ± 0.11 g/fish per body weight [BW]) was significantly larger than those by S1 eels (0.54 ± 0.09 g/fish per BW). There was no difference in fertilization and hatching rates between eggs released by S1 eels and those of S2 eels. These results indicate that the success of induced maturation and spawning in wild female Japanese eels depends on their silvering state, and matured eggs can be obtained efficiently through the use of S2 eels rather than other stages.