Haematological and blood biochemical characteristics of <Emphasis Type="Italic">Glyptosternum maculatum</Emphasis> (Siluriformes: Sisoridae) in Xizang (Tibet)

This study investigated the haematological and blood biochemical characteristics of Glyptosternum maculatum. The haematological and biochemical parameters were measured in 30 adult fish collected from Nyingchi Reach of Yarlung Zangbo River in Tibet. The red blood cell count (RBC), haemoglobin concentration (Hb), haematocrit (Hct), erythrocyte osmotic fragility (maxEof and minEof), the erythrocyte sedimentation rate, mean cell volume (MCV), mean cellular haemoglobin content (MCH), and mean cell haemoglobin concentration (MCHC) were determined. Compared with other Siluriformes fishes, G. maculatum showed similar mean values for Hct, Hb, MCH, and MCHC and had slightly lower RBC and higher MCV. The biochemical parameters were assayed including alanine aminotransferase, aspartate aminotransferase (AST), alkaline phosphatase, total protein, albumin, globulin, albumin/globulin ratio, total bilirubin, direct bilirubin, urea, creatinine, glucose, total cholesterol, and triglyceride. The result showed that the value of AST in G. maculatum was obviously higher than that in Rhamdia quelen as well as in Silurus merdionalis.     

Gastrointestinal and hepatic enzyme activities in juvenile silvery-black porgy (<Emphasis Type="Italic">Sparidentex hasta</Emphasis>) fed essential amino acid-deficient diets

As amino acids (AAs) are vital molecules in the metabolism of all living organisms and are the building blocks of enzymes, a 6-week feeding trial was conducted for determining the influence of dietary essential amino acid (EAA) deficiencies on pancreatic, plasma, and hepatic enzyme activities in silvery-black porgy (initial weight 4.7?±?0.01 g) juveniles. Eleven isoproteic (ca. 47%) and isoenergetic (ca. 20.5 MJ kg^?1) diets were formulated including a control diet, in which 60% of dietary nitrogen were provided by intact protein (fish meal, gelatin, and wheat meal) and 40% by crystalline AA. The other 10 diets were formulated by 40% reduction in each EAA from the control diet. At the end of the experiment, fish fed with threonine-deficient diet showed the lowest survival rate (P?<?0.05), whereas growth performance decreased in fish fed all EAA-deficient diets, although the reduction in body growth varied depending on the EAA considered. Pancreatic enzymes (trypsin, lipase, α-amylase, and carboxypeptidase A) activities significantly decreased in fish fed the EAA-deficient diets in comparison with fish fed the control diet (P?<?0.05). Fish fed with the arginine-deficient diet had the highest plasma and liver alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase levels (P?<?0.05). Plasma and liver lactate dehydrogenase and superoxide dismutase showed the highest and lowest values, respectively, in fish fed the arginine and lysine-deficient diets (P?<?0.05). Plasma metabolites were significantly affected by dietary EAA deficiencies (P?<?0.05). The results of this study suggesting dietary EAA deficiencies led to reduction in growth performance as well as pancreatic and liver malfunction. Furthermore, arginine and lysine are the most limited EAA for digestive enzyme activities and liver health in silvery-black porgy.     

Evaluation of interferon gamma (IFN-γ) of <Emphasis Type="Italic">Labeo rohita</Emphasis> as an immunomodulator: in vitro expression model

Interferon gamma (IFN-γ) or type II interferon is a cytokine that is critical for innate and adaptive immunity against viral and some bacterial and protozoal infections. The importance of IFN-γ in the immune system lies in its ability to inhibit viral replication directly and most importantly from its immunomodulatory effects. Previously, we successfully co-administered IFN-γ along with GAPDH gene of Edwardsiella tarda as bicistronic DNA vaccine in Labeo rohita. In order to ascertain the individual role of IFN-γ, the present study involves cloning and expression of 552-bp IFN-γ open-reading frame (ORF) of L. rohita in striped snakehead (SSN-1) cell line using eukaryotic expression vector system (pQE-TriSystem) followed by transfection in peripheral blood lymphocytes (PBMCs) to evaluate its immunomodulatory ability in comparison to polyinosinic-polycytidylic acid (Poly I:C)-treated PBMCs. The 18.7-kDa protein, expressed in the pQE-IFNγ-transfected SSN-1 cells, reacted with anti-His antibody in Western blot confirming it to be recombinant IFN-γ, whereas the relative expression of IFN-γ, iNOS, Mx, and IL-1β genes in PBMCs was quantified at 24 h and 48 h post treatment by qPCR. The comparative kinetics of all four genes showed significantly (p?<?0.05) high upregulation pattern in both pQE-IFNγ-transfected cell group and Poly I:C-treated cell group demonstrating recombinant IFN-γ as an equally efficient inducer like Poly I:C. Thus, our in vitro experiment results highlight the immunomodulatory potential of recombinant IFN-γ as an analogue to synthetic Poly I:C which warranted future studies to further explore the potential of recombinant IFN-γ as an effective vaccine adjuvant against different microbial invasion.     

Rapid de-adhesion of northern pike <Emphasis Type="Italic">Esox lucius</Emphasis> eggs using sodium hypochlorite

The elimination of egg stickiness is required for effective artificial reproduction of northern pike, but until now, available methods have required at least 40 min. Sodium hypochlorite was tested under laboratory conditions, and exposure to aqueous concentrations of 0.025–0.05% for 40 s effectively eliminated stickiness without adverse effects. Fertilization and hatching rates in laboratory trials were similar to those observed in eggs treated with traditional methods using clay or milk for 40 or 60 min, respectively, as well as those without treatment. Testing using conventional hatchery incubation techniques did not reveal differences in fertilization rates, while the number of hatched larva was significantly higher in eggs treated with sodium hypochlorite vs. clay. Eggs treated with sodium hypochlorite retained transparency, which facilitated monitoring of embryo development.     

Metagenomic analysis using 16S ribosomal RNA genes of a bacterial community in an urban stream,the Tama River,Tokyo

In an effort to determine genus- or species-level taxonomic profiles and diversity of bacterial consortia in the Tama River around urban Tokyo, next-generation sequencing technology targeting a 16S ribosomal RNA (rRNA) gene amplicon was employed. Metagenomic analysis performed by an Ion Personal Genome Machine after sequentially filtering samples through 5-, 0.8- and 0.2-μm filters yielded 1.48 Gb of 16S sequences (average 2.38 M reads/sample). The results indicated that half of the bacterial sequences belonged to Proteobacteria, followed by Bacteroidetes, Actinobacteria and Cyanobacteria. Flavobacterium (Bacteroidetes), possibly including a potential fish pathogen, was the most numerous genera in the Tama River metagenome, and accounted for?~?16% of assigned 16S reads, followed by Mycobacterium. Other dominant bacterial genera including Zoogloea, Sediminibacterium, Hyphomicrobium, Sphingopyxis, Thiothrix and Lysobacter, were thought to be associated with waste water and sludge. MiSeq metagenomic analysis revealed that environmental factors, particularly water temperature, influenced the bacterial composition throughout the year, with a strong negative correlation observed for Proteobacteria and a positive correlation for Bacteroidetes. In terms of bacterial genera, Flavobacterium was positively correlated with temperature, while Polaromonas, Pseudomonas and Bradyrhizobium were negatively correlated with this, suggesting dynamic change in the free-living bacterial population throughout the year and versatile adaptation strategies in relation to environmental factors.     

Effect of sublethal α-cypermethrin exposure on main macromolecules concentration,energy content,and malondialdehyde concentration in free-feeding <Emphasis Type="Italic">Danio rerio</Emphasis> larvae

α-Cypermethrin (Cyp) is a synthetic insecticide used to control pests in agricultural crops and to protect human health against noxious insects; this toxic can reach aquatic systems through ground infiltration or by runoff and could affect the aquatic biota. The present study was aimed at evaluating the acute toxicity of Cyp on zebrafish (Danio rerio) exogenous feeding larvae of 10 and 20 days post-fertilization (dpf), and of sublethal concentrations on only 10-dpf larvae. Proteins, lipids, carbohydrates, glycogen concentration, and total energy contents, as well as malondialdehyde (MDA) quantification, through thiobarbituric acid reactive substances, as a lipid peroxidation biomarker, were assessed in free-feeding larvae exposed to sublethal Cyp concentrations. The LC₅₀ for 10-dpf larvae was 1.94 µg L^?1, and these were more sensitive than 20-dpf larvae (3.56 µg L^?1). The amount of protein, carbohydrates, and glycogen were not significantly affected (p > 0.05), but sublethal Cyp concentrations exposure caused decrement in lipids from 9.05 to 3.74 µg larva^?1, as well as a reduction in MDA and in the total energy content, which affected significantly the development of this fish. Although Cyp is considered an insecticide of reduced residual effect in the environment, the present study revealed that relatively low Cyp concentrations produced significant toxic effects on exogenous feeding fish larvae, a situation that could contribute to increase deaths during this already critical developmental stage in which high mortality is observed frequently.     

Quantitative loop-mediated isothermal amplification method for the detection of <Emphasis Type="Italic">Vibrio nigripulchritudo</Emphasis> in shrimp

Vibrio nigripulchritudo is considered one of the major pathogens threatening shrimp aquaculture. In this study, we developed a novel and highly specific quantitative loop-mediated isothermal amplification (Q-LAMP) assay. A set of four specific primers were designed targeting the V. nigripulchritudo intergenic spacer region. The reaction time and temperature were optimized for 60 min at 63°C. Quantitative analysis was then performed by measuring the turbidity of the reaction solution using a real-time turbidimeter, allowing for quantification of the initial DNA concentration with a sensitivity of 10² copy numbers equivalent to 2.3 colony forming units/ml or 0.3 fg/μl. The LAMP assay was able to specifically detect two representative strains of V. nigripulchritudo, whereas other Vibrio and non-Vibrio species were not amplified. A standard curve was generated for V. nigripulchritudo by plotting the threshold time (T _t) versus the log of bacterial number. A high correlation coefficient (R ² = 0.9749) was observed for the Q-LAMP reaction. In conclusion, Q-LAMP assay is a sensitive, rapid, and simple tool that can be used for the detection and quantification of V. nigripulchritudo in shrimp, thereby facilitating surveillance of vibriosis infection.     

Growth,nutrient utilization,oxidative condition,and element composition of juvenile red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis> fed with fermented soybean meal and scallop by-product blend as fishmeal replacement

A feeding trial was conducted to evaluate the effect of fermented soybean meal and scallop by-product blend (3:2) (FP) on the performance of juvenile red sea bream. Five isocaloric diets were prepared by replacing 0% (FP0), 15% (FP15), 30% (FP30), 45% (FP45), and 60% (FP60) fishmeal (FM) protein with FP, respectively. Triplicate groups of fish (initial mean weight 2.83 g) were fed the test diets for 45 days in a flowthrough seawater system. The results demonstrated that growth rates of fish fed FP0, FP15, and FP30 were similar, and significantly higher (P < 0.05) than those of FP45 and FP60. Nutrient utilization was significantly lower in FP60, with no differences found among the other groups. Dietary heavy-metal contents were affected by inclusion of FP, impacting on whole-body heavy-metal contents. In terms of oxidative stress, fish fed the FP30 diet were in the best condition, since this fish group showed the least oxidative-stressed condition as well as the highest tolerance against oxidation. In conclusion, the approach of utilizing this fermented mixture is promising and it could replace at least 30% FM protein in red sea bream diet without negative effects on performance, body composition or health of fish.