Evolutionary implications of two rainbow trout growth hormone genes

The primary structures of two rainbow trout growth hormone mRNAs (GH1 and GH2) have been deduced by direct sequencing of their respective cDNA clones and portions of the mRNA. Both GH1 and GH2 mRNA contain open reading frames comprised of 630 nucleotides and encode 210 amino acid residues of which 11 are variant. The translated regions of both mRNA are flanked by a short but rather conserved 5′-end, and a relatively long but highly diverged 3′-end. The differences at translated and 3′-untranslated regions suggest that the GH1 and GH2 mRNA originate from different loci. The GH1 and GH2 mRNA are likely transcribed from two distinct loci which were duplicated during tetraploidization of salmonid genome between 50 to 100 million years ago. The GH2 gene has been isolated and sequenced from a rainbow trout genomic library. This gene spans a region of approximately 4 kilobases. The trout GH gene is comprised of 6 exons and 5 introns, in contrast to 5 exons and 4 introns in mammals. The additional intron in the trout gene interrupts the translated regions that are analogous to the last exon of the mammalian counterpart. The alleged internally repeating sequences in mammalian GH, prolactin (Pr1) and placental lactogen (PL) are not observed in the predicted polypeptide sequence of trout GH. In addition, direct repeats that flank exons I, III and V of mammalian GH, Pr1 and PL genes are absent in trout gene. These findings indicate that the rainbow trout GH gene structure does not support the current hypothesis that internally repeated regions in GH, Pr1 and PL arose from a small primordial gene.     

Efficacy of Netminder® silicone release coating for biofouling reduction in bay scallop grow‐out and comparative effects on scallop survival,growth and reproduction

Netminder^®, a water‐based silicone barrier release coating, was highly effective in reducing biofouling of lantern nets during grow‐out/overwintering of bay scallops, Argopecten irradians irradians, in a large‐scale restoration programme in New York, USA. Compared with untreated nets, scallops held in Netminder^®‐coated nets experienced reduced survival after 8 months, probably due to higher loads of epibionts on their shells. However, this was countered by higher reproductive and overall condition of scallops in treated vs. untreated nets; there were no consistent differences in shell growth in different net treatments. Although further testing is recommended under a wider range of conditions, we conclude that for the purposes of our scallop restoration work, coating nets with Netminder^® would eliminate the need for a gear change in spring, thus reducing labour costs; the coating process should be refined so that Velcro closures of nets do not become sealed shut.     

Validation,use, and disadvantages of enzyme-linked immunosorbent assay kits for detection of cortisol in channel catfish,largemouth bass,red pacu,and golden shiners

Measurement of cortisol response is an important tool to asses stress in fisheries research. Radioimmunoassay (RIA) is a common method for the measure of cortisol in fish. Use of enzyme-linked immunosorbent assays (ELISA) to detect cortisol would eliminate health hazards, costs of handling radioisotopes, and the short stability time associated with RIA. Enzyme-linked immunosorbent assays have been used for the determination of cortisol in several fish species. However, the ELISA method of cortisol determination in fish lacks proper validation testing. We conducted validation procedures for multiple commercial cortisol ELISA kits and compared the results to RIA. The assays were tested for four species: (1) channel catfish Ictalurus punctatus, (2) largemouth bass Micropterus salmoides, (3) red pacu Piaractus brachypomus, and (4) golden shiners Notemigonus crysoleucas. We evaluated the ELISA methods against RIA, and determined that at least one kit is suitable (accuracy: mean recovery of spiked samples, 102.8%; reproducibility: interassay coefficient of variation < 10.5% for all species; precision: intra-assay coefficient of variation < 16.8% for all species; linearity: R ² > 0.96 for all species) for the measurement of cortisol response in fish and comparative determination of stress. All of the ELISA assay results varied by more than 10% from the cortisol concentrations detected by the RIA. The high variability of the kit results indicates that commercial ELISA kits could be utilized for qualitative determination of cortisol in fish, but should be fully validated in each laboratory for each species before being used for research.     

Diversity of Flavobacterium psychrophilum and the potential use of its phages for protection against bacterial cold water disease in salmonids

Flavobacterium psychrophilum causes rainbow trout fry syndrome (RTFS) and cold water disease (CWD) in salmonid aquaculture. We report characterization of F. psychrophilum strains and their bacteriophages isolated in Chilean salmonid aquaculture. Results suggest that under laboratory conditions phages can decrease mortality of salmonids from infection by their F. psychrophilum host strain. Twelve F. psychrophilum isolates were characterized, with DNA restriction patterns showing low diversity between strains despite their being obtained from different salmonid production sites and from different tissues. We isolated 15 bacteriophages able to infect some of the F. psychrophilum isolates and characterized six of them in detail. DNA genome sizes were close to 50 Kbp and corresponded to the Siphoviridae and Podoviridae families. One isolate, 6H, probably contains lipids as an essential virion component, based on its chloroform sensitivity and low buoyant density in CsCl. Each phage isolate rarely infected F. psychrophilum strains other than the strain used for its enrichment and isolation. Some bacteriophages could decrease mortality from intraperitoneal injection of its host strain when added together with the bacteria in a ratio of 10 plaque-forming units per colony-forming unit. While we recognize the artificial laboratory conditions used for these protection assays, this work is the first to demonstrate that phages might be able protect salmonids from RTFS or CWD.     

Protection conferred against viral nervous necrosis by simultaneous inoculation of aquabirnavirus and inactivated betanodavirus in the sevenband grouper, Epinephelus septemfasciatus (Thunberg)

An aquabirnavirus (ABV) and a formalin-inactivated betanodavirus [redspotted grouper nervous necrosis virus (RGNNV)] were investigated for their potential to prevent RGNNV-induced viral nervous necrosis (VNN) in the sevenband grouper, Epinephelus septemfasciatus (Thunberg). Three groups of fish were injected intramuscularly with ABV, intraperitoneally with inactivated RGNNV (iRGNNV) or with both ABV and iRGNNV. At 3, 7, 14, 21 and 28 days post-injection (p.i.), fish were challenged by intramuscular injection of RGNNV. Control fish, which received neither ABV nor iRGNNV, showed high mortalities in all RGNNV challenges. Fish that received only ABV exhibited relative percent survival (RPS) of >60 against RGNNV challenges at 3, 7, 14 and 21 days p.i., but not at 28 days p.i., while fish that received only iRGNNV showed significantly higher protection against RGNNV challenges only at 21 and 28 days p.i. In contrast, fish that received both ABV and iRGNNV showed 60 or higher RPS against all RGNNV challenges. Fish inoculated with iRGNNV with or without ABV exhibited similar high titres of neutralizing antibodies to RGNNV at 14, 21 and 28 days p.i. These results indicate that combined inoculation with iRGNNV and ABV conferred both rapid non-specific and delayed specific protection against VNN.     

Salmon testes meal as a functional feed additive in fish meal and plant protein‐based diets for rainbow trout (Oncorhynchus mykiss Walbaum) and Nile tilapia (Oreochromis niloticus L.) fry

Fishery processing by‐products are a large resource from which to produce fishmeal and other products for a variety of uses. In this study, testes meal (TM) produced from pink salmon processing by‐product was evaluated as a functional ingredient in aquafeeds. Nile tilapia and rainbow trout fry were fed five isonitrogenous and isoenergetic experimental diets for 4 and 9 weeks respectively. Two diets were fishmeal‐based (FM) and three were plant protein‐based (PP). Salmon TM was added to the FM and PP diets at 7% to replace 20% of fishmeal protein (FMTM and PPTM respectively). An additional control diet was prepared in which fishmeal was added to the PP diet to supply an equivalent amount of protein as supplied by TM (PPFM). Inclusion of TM in both the FM‐ and PP‐based diets resulted in higher final body weights, although differences were only significant between rainbow trout fed FM or FMTM diets. Similar differences were calculated for other indices of fish performance, e.g. specific growth rate, feed conversion ratio, protein efficiency ratio and protein retention efficiency. Feed intake was significantly higher for fish fed FMTM compared with FM in rainbow trout. For tilapia, final weights were numerically higher, but not significantly different for fish fed diets containing TM compared with non‐TM diets (FM vs. FMTM; PP vs. PPTM). Performance of trout or tilapia fed the PPFM diet did not increase compared with the PP diet. The results indicate that TM addition to both FM and PP diets increased feed intake and also increased metabolic efficiency, demonstrating that TM can be a functional ingredient in aquafeeds.     

Molecular cloning and gene expression of the riboflavin-binding protein in the Nile tilapia, Oreochromis niloticus

Tilapia riboflavin-binding protein (RfBP) cDNA was isolated by library screening. Even though it shows only 31～35% similarity to the reported RfBPs, tilapia RfBP is conserved in amino acid residues that are known to be essential for its protein function. The exclusive expression of RfBP in the brain-pituitary-gonadal axis, as revealed by both RT-PCR and Northern blot, suggested a possible role for RfBP in fish reproduction.     

Inhibition of hirame rhabdovirus growth by RNA aptamers

RNA aptamers are artificial nucleic acids that specifically bind to a wide variety of targets. They are an effective tool for pharmaceutical research and development of antiviral agents. Here, we describe four Hirame rhabdovirus (HIRRV)‐RNA aptamers (H1, H2, H3 and H4) that we obtained from an in vitro process called the systematic evolution of ligands by exponential enrichment (SELEX). The HIRRV‐RNA aptamers specifically bind to HIRRV. Hirame natural embryo (HINAE) cells treated with virus and the RNA aptamer showed a decrease in appearance of cytopathic effect when compared with control (treated only with virus). Rhodovulum sulfidophilum was transformed with genes for the RNA aptamers, and the aptamers were detected in the culture medium, indicating that they were secreted from the cells. Thus, the recombinant R. sulfidophilum might be a powerful tool for the prevention of HIRRV in aquaculture.     

Thirty years of change and the future of Alaskan fisheries: Shifts in fishing participation and diversification in response to environmental,regulatory and economic pressures

Heterogeneity in human responses and decision‐making can contribute to the resilience of social–ecological systems in the face of environmental, political and economic pressures. In fishery systems worldwide, the ability of harvesters to maintain a diverse portfolio of fishing strategies is important for building adaptive capacity. We used a case‐study approach to examine the complexity of factors that inhibit or promote diversification in fisheries of Alaska, one of the major fishing regions of the world. Through a combination of harvest records and literature review, we explored shifts in participation and portfolio diversity in Alaskan fisheries over three decades. The four case‐studies examined the responses of fishers, fleets and communities to multiple, intersecting pressures, including biological declines, market and price dynamics, fishery privatization and the 1989 Exxon Valdez oil spill. These cases illustrate how stressors acting at multiple scales can encourage or constrain opportunities for diversification, and that these opportunities may be spread inequitably across participants. Overall, we found evidence for reduced participation and increasing specialization in Alaskan commercial fisheries. While numerous factors explain these trends, policies like individual quota systems and the increasing cost of entry into fisheries are forcing consolidation at local to regional scales. A portfolio approach to managing fisheries that reduces barriers to diversification and includes broad representation of resource users and communities in management may help to maintain opportunity and choice for fishers.     

Partial cloning of 17B-HSD1 from the Nile tilapia ovary and its expression pattern during spawning cycle

A 548 bp partial cDNA fragment of 17β-hydroxysteroid dehydrogenase (17β-HSD1) was obtained by RT-PCR from the ovary of Nile Tilapia. The expression of 17β-HSD1 was high from 0 to 11 days after spawning, but there was a sharp decline at the day of spawning (day 14) indicating its involvement in ovarian cycle.