Detection of hybrids between masu salmon <Emphasis Type="Italic">Oncorhynchus masou masou</Emphasis> and amago salmon <Emphasis Type="Italic">O. m. ishikawae</Emphasis> occurred in the Jinzu River using a random amplified polymorphic DNA technique

Identification of masu salmon Oncorhynchus masou masou and amago salmon O. m. ishikawae was accomplished using a random amplified polymorphic DNA (RAPD) technique. Of 80 primers screened, three primers amplified subspecies-specific fragments (OPA11-1095 ^*, OPB5-1618^* and OPD5-2038 ^*). Based on fragment patterns, 33 hybrids were detected among 150 upstream-migrating individuals in the Jinzu River, central Japan, a long-time habitat of masu salmon, to which amago salmon were recently introduced. All of the individuals examined in the 2000–2002 cohorts were identified as F ₁ and F _x as well as pure masu and amago forms. These results indicated continuing hybridization, the genetic pollution of the native masu salmon population by amago salmon possibly being a serious problem in the Jinzu River. Both standard length and body weight in F ₁ hybrids tended to be less than in pure masu salmon. However, F _x hybrids showed similar body sizes to masu salmon.     

Farming evaluation of the brownbow triploid hybrid (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis><Emphasis Type="Italic">Salmo trutta</Emphasis>)

Triploid hybrids between female rainbow trout (Oncorhynchus mykiss Walbaum) and male brown trout (Salmo trutta L.) were tested for farming performances, with reference to parental species. The main drawback of hybrids lay in embryonic and larval mortalities, amounting to 60% on average, and displaying a large variability between spawns. Further survival was inferior to that of diploid, but similar to that of triploid rainbow trout. Hybrid body weight was intermediate between weights of rainbow and brown trout of the same age, mainly as a consequence of differences in precocious growth. Analysis of relative growth rates from 6 to 18 months showed that hybrids were surpassed by rainbow controls in common rearing, but not in separate rearing. Hybrid behaviour was similar to that of rainbow trout. These results are discussed in the scope of providing fisheries managers with original and sterile game fishes. An erratum to this article can be found at .     

Spatial learning-induced <Emphasis Type="Italic">egr</Emphasis>-<Emphasis Type="Italic">1</Emphasis> expression in telencephalon of gold fish <Emphasis Type="Italic">Carassius auratus</Emphasis>

The immediate-early gene (egr-1) expression was used to examine the neuron’s response in telencephalon of goldfish during spatial learning in small space. Fishes were pre-exposed in the experimental apparatus and trained to pick food from the tray in a rectangular-shaped arena. The apparatus was divided into identical compartments comprising three gates to provide different spatial tasks. After the fish learned to pass through the gate one, two more gates were introduced one by one. Fish made more number of attempts and took longer time (P < 0.05) to pass through the first gate than the gate two or three. This active learning induces the expression of egr-1 in telencephalon as established by western blot analysis. Subsequently, the fish learn quickly to cross the similar type of second and third gate and make fewer errors with a corresponding decline in the level of egr-1 expression. As the fish learned to pass through all the three gates, third gate was replaced by modified gate three. Interestingly, the level of egr-1 expression increased again, when the fish exhibit a high exploratory behavior to cross the modified gate three. The present study shows that egr-1 expression is induced in the telencephalon of goldfish while intensively acquiring geometric spatial information to pass through the gates.     

Immunostimulatory effect of salmon prolactin on expression of Toll-like receptors in <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis> infected with <Emphasis Type="Italic">Piscirickettsia salmonis</Emphasis>

In aquaculture, antibiotics are the traditional treatment used against bacterial infections. However, their use has increasingly come into question given their effects on fish and, possibly, on human health. Consequently, there is interest in developing alternative treatments aimed at stimulating the innate immune response of fish, which is the first line of defense against pathogens. In relation to this, the Toll-like receptors (TLR) aid in the selective identification of pathogens. The present study evaluated immunostimulatory activity of prolactin (PRL) hormone on expression levels of TLR1, 9, and 22, MyD88, and IL-1β during in vitro infection with the fish pathogen Piscirickettsia salmonis, in primary cultures of Oncorhynchus mykiss head kidney cells. Results indicated that PRL increased expression of TLRs and MyD88 during the first hours of bacterial infection, while a constant increase in expression was found for IL-1β. These findings suggest that PRL indirectly modulates expression of TLRs by activating expression of suppressors of cytokine signaling, thereby regulating immune response over long periods of time during bacterial infection.     

Differences in longitudinal distribution patterns along a Honshu stream of brown trout <Emphasis Type="Italic">Salmo trutta</Emphasis>, white-spotted charr <Emphasis Type="Italic">Salvelinus leucomaenis</Emphasis> and masu salmon <Emphasis Type="Italic">Oncorhynchus masou</Emphasis>

Brown trout Salmo trutta were first introduced into Japan in 1892, and they currently naturally reproduce in several rivers in Honshu and Hokkaido, Japan. Although negative impacts of brown trout introductions on native salmonid fishes have been documented in some Hokkaido rivers, studies of ecological interactions between brown trout and native salmonid fishes on Honshu are limited. In this study, we describe the longitudinal distribution patterns of introduced brown trout, white-spotted charr Salvelinus leucomaenis and masu salmon Oncorhynchus masou in a 4 km stretch of a stream in central Honshu. Underwater observations were conducted in all pools within upstream, middle and downstream sections (190–400 m in length) of this stretch in order to estimate the densities of these species. Only white-spotted charr was observed in the upstream section, while brown trout and masu salmon were observed in the middle and downstream sections. Masu salmon densities, however, were much lower than brown trout densities. In the downstream section, white-spotted charr was absent. These results are consistent with results from previous studies of Hokkaido rivers, where it was found that white-spotted charr in low-gradient areas tend to be displaced by brown trout.     

Quantitative analysis of <Emphasis Type="Italic">Ichthyobodo salmonis</Emphasis> an ectoparasitic flagellate infecting juvenile chum salmon <Emphasis Type="Italic">Oncorhynchus keta</Emphasis> in hatcheries

Ichthyobodosis caused by the ectoparasitic flagellate Ichthyobodo salmonis is a significant cause of mortality in juvenile chum salmon Oncorhynchus keta reared in hatcheries of northern Japan. The present study established a real-time quantitative polymerase chain reaction assay (qPCR) of I. salmonis ribosomal DNA (rDNA) using SYBR Green. This assay allows monitoring of parasite infections for the epidemiological study and control of ichthyobodosis in hatcheries. qPCR showed high reproducibility for measurements between 1.0 and 1.0 × 10⁸ rDNA copy/μl. There was a significant positive correlation between the number of parasites and the amount of I. salmonis rDNA. A survey using the qPCR assay indicated that infection by I. salmonis was present in 23 of 87 hatcheries; parasite loads were estimated to be between 50 and 750 parasites/g juvenile body weight. These results demonstrate that our qPCR assay enables the surveying of juvenile chum salmon reared in hatcheries for infection by I. salmonis.     

Correlation between C-reactive protein level,immunology, and hematology of a <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis> infected with <Emphasis Type="Italic">Lactococcus garvieae</Emphasis>

The aim of this experiment was to evaluate the symptoms, variation, and correlation of hematological factors and some parameters of the innate immune response of the infected rainbow trout (Oncorhynchus mykiss) fish with Lactococcus garvieae bacteria. A total of 270 fish were divided into three groups including infected fish with a high and low concentration of L. garvieae and a control group without infection. The blood and tissue (brain, head, kidney, and spleen) samples were collected from each group (n?=?6 fish) at 0, 3, 14, and 21 days after treatment. The most observed symptoms during infection were lethargy, exophthalmia, and ascites. The mortality rates for the high and low dose-infected fish were defined as 60 and 25%, respectively. The infected groups had significantly (p?<?0.05) higher activity of serum lysozyme, myeloperoxidase, classical and alternative pathways of complement, serum bactericidal effects, and the specific antibody titer. Also, a significant (p?<?0.05) lower hematocrit and hemoglobin levels and higher white blood cell numbers were observed in the infected groups. A significant correlation was observed between the CRP levels and some of the hematological and immunological indices as bactericidal effects, classical complement pathways, lysozyme activity myeloperoxidase activity, white blood cell numbers, and hematocrit levels. The clinical symptoms, immune responses, and hematological indices variation among L. garvieae-infected fish are dependent on the duration and bacterial dose of the infection.     

Probiotic bacteria <Emphasis Type="Italic">Lactobacillus rhamnosus</Emphasis> influences the blood profile in rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis> (Walbaum)

This paper reports the effect of feeding probiotic diets on blood profiles in rainbow trout. Two experiments were performed: in the first, fish of average weight 75 g were offered either a commercial feed or the same incorporated with 10⁹ CFU g⁻¹ of lactic acid bacteria Lactobacillus rhamnosus for 30 days; in the second study performed for a similar duration, fish of average weight 126 g were offered formulated diets that either contained the same bacteria in heat-killed or freeze-dried form (nearly 10¹¹ CFU g⁻¹), or the basal diet without the bacteria. Blood samples were collected at different times after commencement of probiotic feeding to determine the total cholesterol, triglyceride contents, the plasma alkaline phosphatase activity, plasma protein and hematocrit value. The plasma cholesterol significantly increased upon probiotic feeding in the first experiment. A significant elevation (P < 0.05) of plasma cholesterol and triglyceride and alkaline phosphatase activity level was found in the freeze-dried probiotic fed groups at 20 and 30 days postfeeding. This was concomitant with the increased plasma protein and hematocrit values in FD group at 20 and 30 days. Likewise, the heat-killed probiotic fed group registered significantly high values of triglycerides, alkaline phosphatase activity, and plasma protein compared to the control diet fed groups after 20 days of feeding. Thus, alterations in the blood profiles could serve as supplementary information when examining the benefits of probiotics for fish.     

Monitoring of <Emphasis Type="Italic">Francisella noatunensis</Emphasis> subsp. <Emphasis Type="Italic">orientalis</Emphasis> in farmed Nile tilapia (<Emphasis Type="Italic">Oreochromis niloticus</Emphasis>) in Brazil

Francisella noatunensis orientalis is a bacterium that causes emerging bacteriosis in Nile tilapia (Oreochromis niloticus) in many parts of the world, including Brazil. It is a non-motile, Gram-negative, strictly aerobic, facultative intracellular coccobacillus. This species of bacteria is responsible for low to high mortality in fish farms, causing economic losses for fish farmers. This study aimed to detect the presence of F. noatunensis orientalis using qPCR (real-time polymerase chain reaction) and to describe lesions caused by the bacterium in O. niloticus in Brazilian aquaculture. For this purpose, 360 fish from six fish farms (30 per farm) were sampled at two time points (n = 180 per sampling). Necropsies and histopathology were performed for lesion observation, in addition to qPCR and sequencing for detection and identification of Francisella species. Environmental data were collected using a multiparameter sonde YSI EXO2. All measured limnological variables were within the optimum range for cultivation of Nile tilapia. The major lesions present were melanization of the skin, splenomegaly, granulomas, and inflammatory cell responses. The prevalence of francisellosis varied from 0 to 86.66% between time periods and fish farms analyzed, and an outbreak was observed during the second sampling period. This study describes the prevalence of francisellosis in O. niloticus and reports that the lesions found are not exclusively associated with this bacterial disease.     

Wheat grains fermented by fungal mycelia (<Emphasis Type="Italic">Pleurotus ostreatus</Emphasis> or <Emphasis Type="Italic">Lentinus edodes</Emphasis>) as alternative feed ingredients for juvenile rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>)

We investigated the effects of replacing non-fermented wheat grains with wheat grains fermented by fungal mycelia in the diet of juvenile rainbow trout (Oncorhynchus mykiss). We assessed growth performance, feeding parameters, and body composition in three experimental groups (0.33?±?0.01 g, in triplicates of 50 individuals each). The diets for all the groups contained ca. 43% protein and 19% lipids. Experimental diets were made by replacing the 100 g kg^?1 of wheat grains present in the basal diet (CTRL) with the same proportion of wheat grains fermented by Pleurotus ostreatus (PWD) or Lentinus edodes (LWD) mycelium. Fish were fed to apparent satiation twice a day for 56 days. Both, PWD and LWD, significantly increased fish body weight from day 28 onwards. Final body weight was 2.37?±?0.04 g (CTRL), 4.29?±?0.02 g (PWD), and 3.50?±?0.05 g (LWD), and feeding efficiency (%) was increased from 64.5?±?0.7 (CTRL) to 92.5?±?0.5 (PWD) and 84.8?±?1.5 (LWD). The experimental diets also improved nutrient retention efficiency (%): 30.0?±?0.5 (PWD), 27.7?±?1.1 (LWD), and 21.0?±?0.1 (CTRL), for crude protein; 40.3?±?0.6 (PWD), 31.0?±?1.8 (LWD), and 16.1?±?0.7 (CTRL), for ether extract; and 16.1?±?0.1 (PWD), 14.0?±?0.3 (LWD), and 11.6?±?0.6 (CTRL), for phosphorus. Body lipid content was highest for PWD followed by LWD and CTRL (81.4?±?1.4, 63.2?±?2.5, 42.3?±?2.6 g kg^?1, respectively), while viscerosomatic index was lowest for PWD (p?<?0.05). Liver glycogen in LWD and PWD fish (0.62?±?0.10 and 0.21?±?0.08% liver weight) was significantly higher than in CTRL fish (0.05?±?0.01% liver weight). Wheat-mycelium meals appear to be suitable dietary ingredients for improving juvenile rainbow trout growth and nutritional performance. These benefits vary according to the mushroom species used.     

Inflexibility of vertebral number in chum salmon <Emphasis Type="Italic">Oncorhynchus keta</Emphasis>

Chum salmon Oncorhynchus keta eggs from the Chitose and Shiriuchi rivers in Hokkaido, northern Japan, were incubated under identical conditions with the aim of comparing vertebral number (V _N) to that reported archived data from the 1950s. The comparison revealed that mean V _N of salmon from the Shiriuchi River has remained unchanged for 50 years and is higher than that from the Chitose River. The V _N in the Chitose River salmon was found to be significantly higher than that recorded in the archival data. The variation in the mean V _N of chum salmon fry collected during different seasons and years in the Chitose River falls within a range that has remained constant, suggesting that the difference between the values obtained in our study and the archival data may be due to differences in the timing of egg collection. These results indicate that the mean V _N of chum salmon has been stable for a long period. Incubation experiments under identical conditions are a useful tool for examining changes in genetic structure and detecting the essential properties of populations.     

Pathological characteristics of olive flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis> experimentally infected with <Emphasis Type="Italic">Streptococcus parauberis</Emphasis>

Pathological characteristics of olive flounder Paralichthys olivaceus experimentally infected with Streptococcus parauberis were studied. Various stressful conditions, aeration and netting stress in particular, led to induced mortality by S. parauberis. Netting stress-induced mortality was positively correlated to bacterial dose and stressful conditions. Inflammation of the heart and pericarditis was the major pathological change observed in olive flounder experimentally infected with S. parauberis. During the infected period, the number of bacteria in the infected olive flounder was recorded over time. S. parauberis remained in all fish organs tested, especially in the heart and brain.     

Biomass fluctuation of two dominant lanternfish <Emphasis Type="Italic">Diaphus</Emphasis><Emphasis Type="Italic">garmani</Emphasis> and <Emphasis Type="Italic">D. chrysorhynchus</Emphasis> with environmental changes in the East China Sea

Acoustic surveys have been conducted for estimating the biomass of commercially important fish (e.g., anchovy, jack mackerel), lanternfish (Diaphus garmani and D. chrysorhynchus), and pearlside (Maurolicus japonicus) in summer in the East China Sea (ECS) since 1997. The biomass of lanternfish and pearlside was 2.26–19.16 times that of commercially important fish, and these species represented substantial biomass in the ECS. Though there were no correlations between biomass of pearlside and environmental indices, significant correlations between biomass of lanternfish and southern oscillation index (SOI) in March (positive correlation), arctic oscillation (AO) in March (negative) and October (positive), monsoon index (MOI) in February (positive), and Kuroshio flow mass in winter (positive) were observed. Weak AO and strong MOI would cool down the sea temperature and would lead to increased primary and secondary production in the ECS, thereby enhancing larval survival of lanternfish. The SOI would affect the Kuroshio meander in the ECS, and strong SOI and Kuroshio flow mass would transport larvae of lanternfish to the present survey area. This is the first report on the lanternfish standing stock and its fluctuation in the ECS.     

Growth and survival of grouper <Emphasis Type="Italic">Epinephelus</Emphasis><Emphasis Type="Italic">coioides</Emphasis> (Hamilton) larvae fed free-living nematode <Emphasis Type="Italic">Panagrellus redivivus</Emphasis> at first feeding

The free-living nematode, Panagrellus redivivus, was tested as live food for grouper Epinephelus coioides larvae during the first feeding stage. A series of experiments were conducted to determine the acceptability of the free-living nematodes in grouper larvae at first feeding, the optimum nematode density and the response of the larvae to nutritionally enriched nematode. All experiments were conducted in 200-L conical tanks filled with 150-L filtered seawater and stocked at 15 larvae L⁻¹. Duration of feeding experiments was up to day 21 (experiment 1) and 14 days (experiment 2 and 3). Brachionus plicatilis and Artemia (experiment 1) and Brachionus plicatilis alone (experiment 2 & 3) was used as the control treatment. Observations indicated that the grouper larvae readily fed on free-living nematodes as early as 3 days posthatching, the start of exogenous feeding. Optimum feeding density for the larvae was 75 nematodes ml⁻¹. The enrichment of cod liver oil or sunflower oil influenced the total lipids and n-3 highly unsaturated fatty acids of P. redivivus, which in turn influenced those of the grouper larvae, however, growth and survival of the larvae were not affected (P > 0.05). The results from this investigation showed that the nematode, P. redivivus, can be used as first live food for grouper larvae from the onset of exogenous feeding until they could feed on Artemia nauplii.     

Random sequencing of genomic DNA of <Emphasis Type="Italic">Photobacterium damselae</Emphasis> subsp. <Emphasis Type="Italic">piscicida</Emphasis>

ABSTRACT:   Random sequencing of the genomic DNA of Photobacterium damselae subsp. piscicida was conducted. The sequences were assembled into 930 contigs. The total length of these contigs accounts for 37.5% of the genome of P. damselae subsp. piscicida. The contigs contain 2055 open reading frames (ORFs) that have homology with genes in the GenBank database. Furthermore, some of the ORFs have homology with reported virulence related genes, and are classified as encoding colonization factors, exotoxin, and lipopolysaccharide (LPS). Thirty-seven ORFs have homology with colonization factors. In those colonization factors, 27, three, and four ORFs have homology with polar flagellar-related genes, capsule-related genes, and others (accessory colonization factors, superoxide dismutase (Cu-Zn), MshA biogenesis protein, and heme receptor genes), respectively. Five ORFs have homology with exotoxin-related genes (2 hemolysin, phospholipase, hyaluronidase, lysophospholipase L2 genes). Further, six ORFs have homology with LPS-related genes.     

Recent expansion of Northeast Atlantic and Mediterranean populations of <Emphasis Type="Italic">Melicertus</Emphasis> (<Emphasis Type="Italic">Penaeus</Emphasis>) <Emphasis Type="Italic">kerathurus</Emphasis> (Crustacea: Decapoda)

We analysed the genetic diversity of Melicertus kerathurus (Penaeidae), a commercially valuable penaeid shrimp that is distributed in the Mediterranean Sea and eastern Atlantic Ocean. We examined the polymorphism of a 494 bp DNA segment of the mitochondrial COI region in 173 individuals, sampled in nine Mediterranean and two Atlantic samples, covering the whole range of the species from the tropical waters of the Gulf of Guinea to the eastern part of the Mediterranean Sea. The mean nucleotide and haplotype diversities were π = 0.00275 and h = 0.718, respectively, for the global data set, with the highest values occurring in the African samples and the lowest in the Adriatic Sea. A clear sample differentiation was found (F _st = 0.194), but this did not reflect a geographical pattern and there were only faint traces of an Atlantic–Mediterranean subdivision. Mismatch analysis and a high significant negative value of Tajima’s D suggested that M. kerathurus is not at mutation drift-equilibrium, but underwent a recent expansion after a period of low effective sample size. A postglacial recolonisation of the Mediterranean from an Atlantic refuge could be hypothesised based on these data.     

The immune modulatory effect of oregano (<Emphasis Type="Italic">Origanum vulgare</Emphasis> L.) essential oil on <Emphasis Type="Italic">Tilapia zillii</Emphasis> following intraperitoneal infection with <Emphasis Type="Italic">Vibrio anguillarum</Emphasis>

The current study aimed to evaluate the possible effect of Origanum essential oil on innate immune parameters as well as the hematological profiles of Tilapia zillii following challenge with Vibrio anguillarum. Fifty-four of Tilapia zillii weighing 180?±?10.2 g were randomly distributed into three identical closed recirculating seawater systems. The study included three groups (G1, G2, and G3) repeated in triplicates. Fish of the first two groups were fed on a basal diet without herbs, whereas fish of the last group were fed on a basal diet supplemented with Origanum essential oil at concentration 1 g kg^?1 for 15 days. Subsequently, fish of G2 and G3 subjected to a peritoneal inflammation by intraperitoneally injecting V. anguillarum (5.5?×?10⁵ CFU mL^?1), whereas fish of G1 injected with saline and served as control. Fish of all groups were then sampled at 4, 12, and 24 h post injection. No mortalities were observed in both basal and Origanum fed groups. However, some specimens of fish fed basal diet showed dorsal fin erosions, eroded mouth, and detached skin. Although the kinetics of RBCs, WBCs, Hb, and differential leukocyte values remained unchanged in fish fed different diets at the beginning of the trial, significant increases in those values were observed in fish fed Origanum essential oil at 12 and 24 h post injection. Similarly, an augmentation of plasma proteases, antiproteases, and lysozyme activities were recorded in fish fed Origanum essential oil at the same particular sampling points. A significant enhancement in plasma bactericidal capacity was only recorded in fish fed Origanum essential oil at 12 and 24 h post challenge compared to those fed basal diet. In conclusion, Origanum essential oil had a pronounced influence on the innate immunity and increased the fish resistance to V. anguillarum. These data gave insight into the potential use of Origanum in prophylactic strategies against threatening pathogens.