Real-time RT-qPCR assay to detect sequences in the Piscine orthoreovirus-1 genome segment S1 associated with heart and skeletal muscle inflammation in Atlantic salmon

During the last decade, Piscine orthoreovirus was identified as the main causative agent of heart and skeletal muscle inflammation (HSMI) in Atlantic Salmon, Norway. A recent study showed that PRV-1 sequences from salmonid collected in North Atlantic Pacific Coast (NAPC) grouped separately from the Norwegian sequences found in Atlantic Salmon diagnosed with HSMI. Currently, the routine assay used to screen for PRV-1 in NAPC water and worldwide cannot differentiate between the two groups of PRV-1. Therefore, this study aimed at developing a real-time polymerase chain reaction (RT-qPCR) assay to target the PRV-1 genome segments specific for variants associated with HSMI. The assay was optimized and tested against 71 tissue samples collected from different regions including Norway, Chile and both coast of Canada and different hosts farmed Atlantic Salmon, wild Coho Salmon and escaped Atlantic Salmon collected in British Columbia, West Coast of Canada. This assay has the potential to be used for screening salmonids and non-salmonids that may carry PRV-1 potentially causing HSMI.     

Virucidal effects of various agents—including protease—against koi herpesvirus and viral haemorrhagic septicaemia virus

In a search for alternative, environmentally friendly and effective disinfecting agents, a commercially available protease—Neutrase^®—was tested in this work for inactivation of koi herpesvirus (KHV) and of viral haemorrhagic septicaemia virus (VHSV). For comparison, the stability of these viral pathogens in similar configurations at various pH values and concentrations of peracetic acid or quicklime, typically used for disinfection, was tested. Therefore, virus suspensions were incubated with various concentrations of different agents for 24 hr and the titre of the remaining infectious particles was determined by virus titration. Furthermore, the treatment of both viruses, with the agents at concentrations that were previously appointed as effective, was also examined in the presence of solid material (quartz sand). All procedures investigated in this study, including the protease treatment, were able to reduce the titre of KHV and VHSV below the detection limit of the titration. Although further studies are necessary, this is the first report of the application of a protease for the inactivation of the selected fish pathogens, demonstrating the great potential of the latter for disinfection.     

Assessment of Bioactive Potential of Aqueous Protein Extracts from Diatoms Nitzschia laevis,Spirulina platensis,and Chlorella vulgaris

The bioactivities of protein extracts from Nitzschia laevis, Spirulina platensis, and Chlorella vulgaris were evaluated in vitro. Free radical scavenging potential, reducing power, oxygen radical absorbance capacity, superoxide anion radical, xanthine oxidase inhibition, and potential to inhibit angiotensin 1-converting enzyme (ACE) and acetylcholinesterase enzyme (AChE) were studied. Nitzschia protein extracts showed the highest 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activities, reducing power, and ACE inhibitory activity. Spirulina’s proteins showed the highest 2,2′-Azino-bis (3 ethylbenzothiazoline-6-sulfonic acid) (ABTS) scavenging activity, xanthine oxidase, and AChE inhibition activities. Aqueous protein extracts from Nitzschia, Spirulina, and Chlorella showed in vitro antioxidant, anti-ACE, and anti-AChE activities, suggesting possible new sources of bioactive proteins of different phyla with nutraceutical and pharmaceutical potentials.     

Essential amino acids supplementation to practical diets affects growth,feed utilization and glucose metabolism‐related signalling molecules of blunt snout bream (Megalobrama amblycephala)

Six diets were formulated to investigate the success of fish meal (FM) replacement by plant proteins; diet 1 reflected a commercial feed (8% FM), diet 3 contained 4% FM, and diet 5 was devoid of FM. Whereas, diet 2, diet 4, and diet 6 reflected diet 1, diet 3, diet 5, respectively, and supplemented with essential amino acid (EAA). At the end of 8‐week trial, there was no significant difference in survival rate. Significantly higher final weight, weight gain rate, and specific growth rate were recorded in the group fed diet 2 compared with the other treatments (except diet 4) (p < 0.05). Feed conversion ratio of fish fed diet 2 was significantly lower than those fed diets 1, 3, and 5 (p < 0.05). The lowest feed intake and highest protein efficiency rate were found in fish fed diet 2 (p < 0.05). There was no significant difference in whole body compositions between treatments. Plasma aspartate transaminases, alanine aminotransferase, and glucose were significantly affected by dietary treatments (p < 0.05), while plasma protein and albumin contents were not influenced by the treatments. The relative expression of target of rapamycin (TOR) and phosphatidylinositol 3‐kinase, regulatory subunit 1 (alpha) (PIK3R1) in fish fed diet 3 (4% FM) were significantly down‐regulated compared with those fed diet 6 for TOR and diets 4 and 6 for PIK3R1 (p < 0.05). Insulin receptor substrate 1 (IRS‐1) and janus kinase 3 (JAK3) expressions were fluctuated, with the higher levels in fish fed diets 4 and 6. In conclusion, the findings of this study indicate that plant protein mixture supplemented with EAA could be used to substitute FM in practical diet for Megalobrama amblycephala.     

Dietary supplementation with low molecular weight sodium alginate improves growth,haematology, immune reactions and resistance against Aeromonas hydrophila in Clarias gariepinus

A few studies have illustrated the effects of sodium salt derived from alginic acid on different fish species. However, little is known about the effect of sodium alginate on catfish (Clarias gariepinus). Therefore, this study was performed to assess the use of low molecular weight sodium alginate ( LMWSA) in C. gariepinus. A total of 180 apparently healthy C. gariepinus with a mean body weight of 45 g were randomly divided into three equal groups (D1, D2 and D3). D1 the control group received a control diet, while D2 and D3 received 1% and 3% LMWSA, respectively, for 8 weeks. A challenge test against Aeromonas hydrophila was performed on 15 randomly selected catfish for 15 days. At the end of the experiment, catfish in D3 that received a diet of 3% LMWSA showed significant increases in the final body weight, weight gain and thermal‐unit growth coefficient compared with those in D2 and D1. There was a significant decrease in the erythrogram in D1 after the 4‐day pathogen challenge. A leucogram revealed leucocytosis, heterophilia and lymphocytosis in catfish in D2 and D3 compared with those in D1. After the 4‐day challenge, the following changes took place: lysozyme, nitric oxide, phagocytic activity and the respiratory burst were significantly elevated in catfish that received LMWSA and were more pronounced in D3 than in D1. The mortalities of catfish have been stopped after pathogen challenge from 8‐day in D1 and D2 where at 6‐day in D3. Thus, administration of 1% and 3% LMWSA enhances the growth, immune response and resistance of C. gariepinus against A. hydrophila.     

油炸糯米糕营养效应评价及其对血脂健康的影响

为研究食用油炸糯米糕的整体营养健康效应及其对血脂健康的影响,自制纯油炸糯米糕和蒸糯米糕,选用健康雄性小白鼠,分别用油炸糯米糕外壳层、中间层、油炸糯米糕全糕及蒸糯米糕进行6周连续灌胃试验,饲料、葡萄糖作为对照组。采用模拟BDI-GS体系评价法进行营养效价、健康效应评定及试剂盒法对小鼠血脂指标测定,并计算相应指标数据,做图对比分析。结果表明,外壳层组平均日采食量为（5.70 ± 0.56）g,其余各组为6.80 g左右,食用油炸外壳层组的小鼠采食量及体重显著下降;营养健康方面,油炸糯米糕和蒸糯米糕相比无显著差异,各试验样品的累加积分均大于8.0,基本益于机体整体营养和健康;血脂方面,油炸糯米糕全糕会提高小鼠血脂中总胆固醇（TC）和低密度脂蛋白胆固醇（LDL-C）含量,油炸糯米糕外壳层会提高总胆固醇（TC）、甘油三酯（TG）和低密度脂蛋白胆固醇（LDL-C）含量。     

Effects of commercial thinning,log position in the stem,and cutting width on the surface quality of cants produced by a chipper-canter

Fifteen stems of jack pine (Pinus banksiana Lamb.) of 3 commercial thinned plots (control, moderate, and intensive thinning) and 15 stems of black spruce (Picea mariana (Mill.) B.S.P), both coming from the Abitibi-Témiscamingue region – Canada, were cross-cut into three 2.4?m length sections: bottom, middle, and top logs. Logs were processed with a chipper-canter at three cutting widths (12.7, 19.1, and 25.4?mm), producing chips and a three-faced cant. The middle section of the cant was used to evaluate surface quality across the grain on each face. Roughness and waviness parameters and depth of torn grain were recorded. Knot characteristics were assessed in the three cant faces. Poorer surface quality was found in the lower part compared to the upper part of the cant for both species. At larger cutting widths, jack pine logs coming from a natural stand showed lower surface quality compared to logs from thinned stands. Black spruce waviness increased with the cutting width and stem height. These results were attributed to the increase of forces and vibration when cutting at larger cutting widths, which was worsened by the presence of bigger, more numerous knots at the control plot and in the top logs. Black spruce had deeper torn grain compared to jack pine. Their differences in knot characteristics resulted in a maximum torn grain depth favored by the presence of more knots rather than by bigger knot size. Other knot characteristics, such as the position of the knots in the cant face, the insertion angle of the branch and the distribution of the knots around the log, should be studied to better understand the relationship between torn grain formation and knottiness.     

Molecular characterization and identification of facilitative glucose transporter 2 (GLUT2) and its expression and of the related glycometabolism enzymes in response to different starch levels in blunt snout bream (<Emphasis Type="Italic">Megalobrama amblycephala</Emphasis>)

Facilitative glucose transporters (GLUT) are transmembrane transporters involved in glucose transport across the plasma membrane. In this study, blunt snout bream GLUT2 gene was cloned, and its expression in various tissues and in liver in response to diets with different carbohydrate levels (17.1; 21.8; 26.4; 32.0; 36.3; and 41.9% of dry matter). Blunt snout bream GLUT2 was also characterized. A full-length cDNA fragment of 2577 bp was cloned, which contains a 5′-untranslated region (UTR) of 73 bp, a 3′-UTR of 992 bp, and an open reading frame of 1512 bp that encodes a polypeptide of 503 amino acids with predicted molecular mass of 55.046 kDa and theoretical isoelectric point was 7.52. The predicted GLUT2 protein has 12 transmembrane domains between amino acid residues at 7–29; 71–93; 106–123; 133–155; 168–190; 195–217; 282–301; 316–338; 345–367; 377–399; 412–434; and 438–460. Besides, the conservative structure domains located at 12–477 amino acids belong to the sugar porter family which is the major facilitator superfamily (MFS) of transporters. Blunt snout bream GLUT2 had the high degree of sequence identity to four GLUT2s from zebrafish, chicken, human, and mouse, with 91, 63, 57, and 54% identity, respectively. Quantitative real-time (qRT) PCR assays revealed that GLUT2 expression was high in the liver, intestine, and kidney; highest in the liver and was regulated by carbohydrate intake. Compared with the control group (17.1%), fed by 3 h with higher starch levels (32.0; 36.3; and 41.9%), increased plasma glucose levels and glycemic level went back to basal by 24 h after treatment. Furthermore, higher dietary starch levels significantly increase GLUT2, glucokinase (GK), and pyruvate kinase (PK) expression and concurrently decrease phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6P) mRNA levels (P?<?0.05), and these changes were also back to basal levels after 24 h of any dietary treatment. These results indicate that the blunt snout bream is able to regulate their ability to metabolize glucose by improving GLUT2, GK, and PK expression levels and decreasing PEPCK and G6P expression levels.     

Influence of different microalgal diets on gonadal development of the carpet shell clam <Emphasis Type="Italic">Ruditapes decussatus</Emphasis> broodstock

Studies considering broodstock conditioning of Ruditapes decussatus mostly used flagellates and diatoms for feeding, mainly Isochrysis galbana and Chaetoceros calcitrans, respectively. The present study evaluated the effect of different microalgal diets on gonadic maturation of R. decussatus including three microalgae species (C. calcitrans, Tetraselmis suecica, and Nannochloropsis oculata) which were used to prepare six diets: three monospecific and three bispecific mixed diets at a proportion of 1:1. Clams were daily fed at a ratio of 1% dry weight of algae/live weight of clam and unfed treatment was used as control. Results showed that clams fed C. calcitrans alone or mixed with T. suecica attained earlier full maturity with the highest response to spawning induction (after 34 conditioning days) and largest proportion in the partial spawning phase for both, followed by T. suecica alone after 48 days. In contrast, N. oculata alone or mixed diets demonstrated poor performance for gonadal maturation. Unfed treatment did not show any sign of maturation. The results indicated that T. suecica has high conditioning potential like C. calcitrans and their mixture gave the highest benefit. Therefore, T. suecica can be recommended for R. decussatus broodstock conditioning.