Comparative pathogenicity study of ten different betanodavirus strains in experimentally infected European sea bass,Dicentrarchus labrax (L.)

Viral encephalopathy and retinopathy (VER), otherwise known as viral nervous necrosis (VNN), is a severe pathological condition caused by RNA viruses belonging to the Nodaviridae family, genus Betanodavirus. The disease, described in more than 50 fish species worldwide, is considered as the most serious viral threat affecting marine farmed species in the Mediterranean region, thus representing one of the bottlenecks for further development of the aquaculture industry. To date, four different genotypes have been identified, namely red‐spotted grouper nervous necrosis virus (RGNNV), striped jack nervous necrosis virus (SJNNV), tiger puffer nervous necrosis virus and barfin flounder nervous necrosis virus, with the RGNNV genotype appearing as the most widespread in the Mediterranean region, although SJNNV‐type strains and reassortant viruses have also been reported. The existence of these genetically different strains could be the reason for the differences in mortality observed in the field. However, very little experimental data are available on the pathogenicity of these viruses in farmed fish. Therefore, in this study, the pathogenicity of 10 isolates has been assessed with an in vivo trial. The investigation was conducted using the European sea bass, the first target fish species for the disease in the Mediterranean basin. Naive fish were challenged by immersion and clinical signs and mortality were recorded for 68 days; furthermore, samples collected at selected time points were analysed to evaluate the development of the infection. Finally, survivors were weighed to estimate the growth reduction. The statistically supported results obtained in this study demonstrated different pathogenicity patterns, underlined the potential risk represented by different strains in the transmission of the infection to highly susceptible species and highlighted the indirect damage caused by a clinical outbreak of VER/VNN.     

Odor Modification in Salmon Hydrolysates Using the Maillard Reaction

The aim of this work was to study the effect of adding sugar during proteolysis to promote the Maillard reaction and mask the initial fish odor and off-flavors generated. An experimental design, based on the Doehlert plan, was used to study the influence of hydrolysis conditions (time, temperature, sugar, and antioxidant addition) on the odor characteristics of hydrolysates, soluble protein levels, and amino acid content. Results showed that the lowest level of sugar (10 g of D-xylose added to 1 kg of by-products) was enough to develop a grilled odor in hydrolysates. In the hydrolysis conditions used—i.e., enzyme inactivation at 95°C for 30 min—hydrolysis temperature had no effect on grilled odor production but significantly affected the soluble protein fraction, as did hydrolysis time. Soluble protein content and essential amino acid content increased with the enzymatic reaction but were not modified by adding sugar. Hydrolysis conditions that promote Maillard reactions while keeping a nutritional balance have been identified.     

Species identification method for <Emphasis Type="Italic">Scombrops boops</Emphasis> and <Emphasis Type="Italic">Scombrops gilberti</Emphasis> based on polymerase chain reaction-restriction fragment length polymorphism analysis of mitochondrial DNA

ABSTRACT:   Gnomefish Scombrops boops and Scombrops gilberti are commercially important fishes in Japan, but these species are often confused in the markets because of their morphological similarity. To identify these two species, we performed nucleotide sequencing and restriction fragment length polymorphism (RFLP) analysis on 16S ribosomal RNA (rRNA) gene and the control region in mitochondrial DNA. Five and 12 nucleotide substitutions were observed between species in the 777-bp 16S rRNA gene and 471-bp control region, respectively. Diagnostic restriction sites for discriminating between S. boops and S. gilberti were found in the 16S rRNA gene, but not in the control region. Polymerase chain reaction (PCR)–RFLP analysis using two enzymes, Eco NI and Mva I, clearly discriminated between S. boops and S. gilberti identified by meristic characters. The PCR–RFLP analysis identified most of the 168 Scombrops young caught in the coastal waters of the Izu and Miura peninsulas as S. boops , suggesting that S. gilberti juveniles are rare in this area.     

Ascorbic acid status as affected by dietary treatment in the Siberian sturgeon (Acipenser baeri Brandt): tissue concentration,mobilisation and L-gulonolactone oxidase activity

A study was conducted to evaluate tissue storage and mobilisation of L-ascorbic acid (AA) in the Siberian sturgeon (Acipenser baeri) fed three different experimental diets. The three treatments consisted of a diet devoid of vitamin C (diet A0) and two diets supplemented with equivalent of 300 mg AA kg^–1 in the form of either silicone-coated ascorbic acid (diet SC) or of ascorbyl-2-polyphosphate (diet AP). During the first phase (4 months) of the trial, six batches of 130 Siberian sturgeon (initial body weight: 25.5±0.5 g) each were fed one of the three diets in duplicate. During the second phase (3 months), fish from groups SC and AP were switched to diet A0 and those fed diet A0 during the first phase were switched to diet SC. Irrespective of the dietary treatment, growth rates were not significantly different from each other. At the end of phase I, in all tissues studied, total ascorbic acid (TAA) concentrations were higher in Siberian sturgeon fed diet AP than in the other two groups. During phase II, tissue ascorbate depletion was also higher in the AP group than in the other two groups. Transfer of the AA-free diet fed group onto a diet supplemented with 300 mg AA kg^–1 (diet SC) led to a slight increase in the TAA concentrations in all tissues. Blood plasma tyrosine concentrations were not significantly different between the three groups. Whole-body collagen levels were affected by dietary AA levels or forms at the end of phase I; the differences were not significant at the end of phase II. Muscle collagen levels were slightly affected. L-Gulonolactone oxidase activity was found in the kidney of Siberian sturgeon, but not in the liver. The ascorbyl-2-polyphosphate appears to be either better utilised by Siberian sturgeon, like in many other teleosts, or more stable than the silicone-coated AA during food processing and storage. Presence of L-gulonolactone oxidase activity in Siberian sturgeon kidney combined with the absence of gross scorbutic signs in AA-free diet fed groups expressing very good growth rates suggested no need of dietary AA byA. baeri.     

Recreational fishery as a conservation tool for endemic Dolly Varden <Emphasis Type="Italic">Salvelinus malma miyabei</Emphasis> in Lake Shikaribetsu,Japan

Recreational angling is a popular leisure activity globally and can be a threat or useful conservation tool depending on the management strategy. Miyabe charr Salvelinus malma miyabei is an endemic fish that inhabits Lake Shikaribetsu, and a management program was established to harmonise recreational fishery and conservation. To examine the suitability of this program, population assessment was conducted with the cooperation of anglers. The population size in 2014 as assessed by tag-and-release angling was estimated to be 105,300 [95% confidence interval (CI) 37,300–178,600], much higher than the reported estimate in 1995. Further, angling mortality was estimated to be quite low. Moreover, no decreasing population trend was detected on analysing 8 years of anglers’ catch data. Consequently, angling has facilitated stock assessments of Miyabe charr under the current program. This case is a good example of recreational angling acting as a conservation tool under appropriate management.     

Digestibility of the defatted microalgae Nannochloropsis sp. and Desmodesmus sp. when fed to Atlantic salmon,Salmo salar

The aim of the study was to investigate the apparent digestibility coefficient (ADC) of defatted biomass derived from microalgae Nannochloropsis sp. and Desmodesmus sp. when fed to Atlantic salmon postsmolts in seawater. Two experiments, one employing cold‐pelleted and the other employing extruded pellets, were carried out to determine the ADC of dry matter (DM), protein, ash and energy. The test feeds consisted of a fishmeal‐based reference feed and the algal biomass, at a ratio of 70:30, and yttrium oxide as the inert marker. The ADC of DM and protein in fish fed microalgae‐incorporated cold‐pelleted feeds were significantly higher for Nannochloropsis sp. compared with Desmodesmus sp. The Nannochloropsis sp. inclusion in extruded feeds also led to higher ADCs of DM and energy compared with Desmodesmus sp. The extrusion processing significantly increased the ADC of ash in both defatted microalgae biomass, as well as ADCs of DM in Nannochloropsis and protein in Desmodesmus sp. In conclusion, the microalga Nannochloropsis sp. was more digestible than Desmodesmus sp., and extrusion processing can be used as a means to improve digestibility of certain nutrients.     

Regulation of growth, fatty acid composition and delta 6 desaturase expression by dietary lipids in gilthead seabream larvae (Sparus aurata)

The Δ6 and Δ5 desaturases and elongases show only very limited activity in marine fish, and little is known of the possibility of enhancing Δ6 desaturase gene expression in these fish. The use of plant oils in marine fish diets is limited by their lack of n−3 highly unsaturated fatty acids (HUFA) despite an abundant content of the 18C fatty acid precursor linoleic and α-linolenic acids. The objective of the present study was to determine the ability of larval gilthead seabream to utilize vegetable oils and assess the nutritional regulation of Δ6 desaturase gene expression. Seventeen-day-old gilthead seabream larvae were fed during a 17-day period with one of four different microdiets formulated with either sardine fish oil (FO), soybean, rapeseed or linseed oils, respectively, or a fifth diet containing defatted squid meal and linseed oil. Good larval survival and growth, both in terms of total length and body weight, were obtained by feeding the larvae either rapeseed, soybean or linseed oils. The presence of vegetable oils in the diet increased the levels of 20:2n−9 and 20:2n−6, 18:2n−9, 18:3n−6, 20:3n−6 and 20:4n−6, in larvae fed rapeseed and soybean oils in comparison to those fed FO. In addition, a sixfold increase in the relative expression of Δ6 desaturase-like gene was found in larvae fed rapeseed and soybean oils, denoting the nutritional regulation of desaturase activity through its gene expression in this fish species. However, feeding linseed oil did not increase the expression of the Δ6 desaturase gene to such a high extent.     

Utilization of monounsaturated fatty acid (18:1n-9, oleic acid) by freshwater prawn Macrobrachium rosenbergii (de Man) juveniles

The utilization of oleic acid as an energy source and the effects of oleic acid levels and/or dietary soy bean lecithin (SBL) on oleic acid utilization, growth and survival, and lipid class and fatty acid compositions of freshwater prawn, Macrobrachium rosenbergii (de Man), juveniles were determined.
Increase in levels of dietary oleic acid from 10 to 80 g kg⁻¹ significantly ( P ≤ 0.05) reduced growth, survival and feed conversion efficiency of M. rosenbergii juveniles during the 40-day feeding period. Inclusion of 20 g kg⁻¹ SBL had no significant effect ( P ≥ 0.05) on growth and survival, nor was there any interactive effect between dietary SBL and oleic acid levels.
Body fatty acid profile of prawns reflected dietary fatty acid quantity. The fatty acid composition of prawns fed diets containing 80 g kg⁻¹ oleic acid had tremendously high proportions of oleic acid. Polar lipids, mostly phosphatidylcholine (PC), constituted the bulk of the extracted total lipids. Prawns fed with SBL had significantly ( P ≥ 0.05) higher PC content.
Oleic acid was metabolized for energy by M. rosenbergii juveniles at the same rate regardless of dietary level of SBL and/or oleic acid. Expired ¹⁴CO₂ accounted for half of the ingested radioactivity 48 h after feeding with labelled diets. No significant difference in the amount of ¹⁴CO₂ expired by prawns fed the labelled test diets was found. Per cent radioactivity ingested and absorbed into the body was also not significantly different in prawns of the different dietary treatments.     

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.     

Differences in uptake and killing of pathogenic and non‐pathogenic bacteria by haemocyte subpopulations of penaeid shrimp,Litopenaeus vannamei, (Boone)

Phagocytosis is an important function of both invertebrate and vertebrate blood cells. In this study, the phagocytic activity of haemocyte subpopulations of penaeid shrimp, Litopenaeus vannamei, (Boone), against pathogenic and non‐pathogenic particles was investigated in vitro. The haemocytes of penaeid shrimp were firstly separated by centrifugation on a continuous density gradient of iodixanol into four fractions with five subpopulations (sub), of which sub 1 (hyalinocytes) and sub 4 (semi‐granulocytes) have the main function in phagocytosis of both pathogenic and non‐pathogenic bacteria as well as fluorescent polystyrene beads. It was found that these haemocyte subpopulations engulfed virulent Vibrio campbellii and Vibrio harveyi at a higher rate than non‐virulent Escherichia coli and polystyrene beads. When these bacteria were mixed with shrimp haemocyte subpopulations and incubated for 180 min, the percentage of viable intracellular V. campbellii (25.5 ± 6.0%) recovered was significantly higher than the percentage recovered from V. harveyi (13.5 ± 1.1%). No viable intracellular E. coli was observed in this study. In contrast to V. harveyi and E. coli, V. campbellii containing endosomes did not acidify in time. Incubation of haemocyte subpopulations with the most virulent V. campbellii strain resulted in a significant drop in haemocyte viability (41.4 ± 6.3% in sub 1 and 30.2 ± 15.1% in sub 4) after 180 min post‐inoculation in comparison with the less virulent V. harveyi (84.1 ± 5.6% in sub 1 and 83.4 ± 4.1% in sub 4) and non‐virulent E. coli (92.7 ± 2.8% in sub 1 and 92.3 ± 5.6% in sub 4) and polystyrene beads (91.9 ± 1.6% in sub 1 and 84.4 ± 3.4% in sub 4). These findings may be a valuable tool for monitoring shrimp health and immunological studies.