Crystal structure and nucleotide sequence of an anionic trypsin from chum salmon (Oncorhynchus keta) in comparison with Atlantic salmon (Salmo salar) and bovine trypsin

The nucleotide sequence and crystal structure of chum salmon trypsin (CST) are now reported. The cDNA isolated from the pyloric caeca of chum salmon encodes 222 amino acid residues, the same number of residues as the anionic Atlantic salmon trypsin (AST), but one residue less than bovine beta-trypsin (BT). The net charge on CST determined from the sum of all charged amino acid side-chains is -3. There are 79 sequence differences between CST and BT, but only seven sequence differences between CST and AST. Anionic CST isolated from pyloric caeca has also been purified and crystallized; the structure of the CST-benzamidine complex has been determined to 1.8A resolution. The overall tertiary structure of CST is similar to that of AST and BT, but some differences are observed among the three trypsins. The most striking difference is at the C terminus of CST, where the expected last two residues are absent. The absence of these residues likely increases the flexibility of CST by the loss of important interactions between the N and C-terminal domains. Similarly, the lack of Tyr151 in CST (when compared with BT) allows more space for Gln192 in the active site thereby increasing substrate accessibility to the binding pocket. Lys152 in CST also adopts the important role of stabilizing the loop from residue 142 to 153. These observations on CST provide a complementary view of a second cold-adapted trypsin, which in comparison with the structures of AST and BT, suggest a structural basis for differences in enzymatic activity between enzymes from cold-adapted species and mammals.     

Purification and characterization of biliverdin IXalpha from Atlantic salmon (Salmo salar) bile

Biliverdin IX was purified from the bile of Atlantic salmon (Salmo salar) using a silica gel (Wakogel C-200) column. The yield was 49.5 mg per 100 ml of fresh bile and purity 95.3%. The biliverdin IX in the bile was quite stable when the bile was frozen at –80°C for a period of 40 days. However, 7.1% of the biliverdin IX was lost when the bile was stored at 4°C for 20 days. The purified biliverdin IX appeared as a single spot with R_f value of 0.25-0.27 on thin layer chromatography (TLC) and one main peak on high performance liquid chromatography (HPLC) at 436 or 650 nm. When the biliverdin IX was subjected to enzymic reduction with highly purified biliverdin reductase, two clear isobestic points were seen, at 384 and 670 nm. When the products of the reaction with biliverdin IX were extracted in butanol after completion of the reaction, one absorbance peak was observed at 468 nm. The time course of the reduction of biliverdin IX to bilirubin IX catalyzed by biliverdin reductase depended on reduced pyridine nucleotide. The time course of the NADPH-dependent reaction is different from that of the reaction with NADH. In the reduction of biliverdin IX , per mole of biliverdin IX reduced or per mole of bilirubin IX formed 1 mole of reduced pyridine nucleotide was consumed in both the NADH and NADPH systems.     

Factors affecting the outcome of territorial contests between hatchery and naturally reared coho salmon parr in the laboratory

In aquarium experiments using coho salmon as a model species, prior residents dominated intruders of the same size but intruders with a 6% length advantage were equally matched against prior residents. Prior winning experience (distinct from individual recognition) also strongly influenced competitive success and overcame a prior residence effect. Coho salmon reared in a hatchery dominated size-matched fish from the same parental population reared in a stream. Hatchery-reared salmon also dominated naturally spawned salmon, even when the wild salmon were prior residents. Thus the combined effects of greater size and rearing experience of hatchery-produced salmon were sufficient to overcome a wild salmon's advantage of prior residence. Efforts to rehabilitate salmonid populations must consider such behavioural interactions if displacement of wild fish is to be prevented.     

The impact of a brominated flame retardant on smoltification and olfactory function in Atlantic salmon (Salmo salar L.) smolts

Despite a reduction in exploitation of salmon stocks throughout the NE Atlantic, there continues to be a decline in many populations. The factors regulating these populations remain poorly understood, although there is evidence that environmental conditions experienced in freshwater can effect survival in the marine environment. The aim of this study was to investigate the impact of a brominated flame retardant (hexabromocyclododecane, HBCD) on the parr-smolt transformation in juvenile salmon. It is during this pre-adaptive period to marine life that olfactory imprinting to the natal river is considered to occur. Fish were exposed to low levels of HBCD for 30 days over the peak smoltification period in freshwater, and then transferred to clean seawater for 20 days. Fish were sampled weekly to assess changes in some of the physiological parameters associated with smoltification, and olfactory response to conspecific smolt urine was measured using an electro-olfactogram (EOG). Exposure to HBCD did not affect seawater adaptability, although there was some disruption of plasma thyroid hormone levels, as well as a reduction in olfactory function to conspecific smolt urine. The results are discussed in relation to the marine survival and successful homing of adult salmon.     

Extrapolating Growth Reductions in Fish to Changes in Population Extinction Risks: Copper and Chinook Salmon

Fish commonly respond to stress, including stress from chemical exposures, with reduced growth. However, the relevance to wild populations of subtle and sometimes transitory growth reductions may not be obvious. At low-level, sustained exposures, Cu is one substance that commonly causes reduced growth but little mortality in laboratory toxicity tests with fish. To explore the relevance of growth reductions under laboratory conditions to wild populations, we (1) estimated growth effects of low-level Cu exposures to juvenile Chinook salmon (Oncorhynchus tshawytscha), (2) related growth effects to reduced survival in downriver Chinook salmon migrations, (3) estimated population demographics, (4) constructed a demographically structured matrix population model, and (5) projected the influence of Cu-reduced growth on population size, extinction risks, and recovery chances. Reduced juvenile growth from Cu in the range of chronic criteria concentrations was projected to cause disproportionate reductions in survival of migrating juveniles, with a 7.5% length reduction predicting about a 23% to 52% reduction in survival from a headwaters trap to the next census point located 640 km downstream. Projecting reduced juvenile growth out through six generations (～30 years) resulted in little increased extinction risk; however, population recovery times were delayed under scenarios where Cu-reduced growth was imposed.     

Whole body elemental and proximate composition of Atlantic salmon (Salmo salar) during the life cycle

Atlantic salmon ( Salmo salar ) raised under production conditions were sampled periodically during their life cycle, in fresh and salt water, to determine their proximate and elemental composition. The parameters changed in relation to life cycle stage and fish size, and were influenced by whether the fish were in salt or freshwater. Measurements presented can be used to assist in the formulation of feeds, and to determine the health and nutritional status of commercially reared Atlantic salmon.     

Genetic structure of masu salmon (Oncorhynchus masou) populations in Hokkaido, northernmost Japan, inferred from mitochondrial DNA variation

Genetic structure of masu salmon Oncorhynchus masou populations in Hokkaido was examined by analysing mtDNA NADH dehydrogenase subunit 5 gene (561 bp) of 382 individuals collected from 12 rivers, in which there were no records of artificial release. Analysis of molecular variance showed that between groups level and between populations within-group level explained each c. 10% of genetic variance. In neighbour-joining tree, four populations located in southern Hokkaido were clustered into a single group; however, other populations did not form any clear clusters. Fu's F _S, Tajima's D and a mismatch distribution test indicated a sudden expansion of population in the entire population of Hokkaido and the northernmost population of Chiraibetsu, which was genetically close to the southern Hokkaido group. The Sea of Japan and southern rivers, including those of southern Hokkaido, seem to have served as refugia for masu salmon during glacial periods, and their dispersal and straying in interglacial periods affected the genetic structure of masu salmon populations in Hokkaido.     

Stream flow, salmon and beaver dams: roles in the structuring of stream fish communities within an anadromous salmon dominated stream

The current paradigm of fish community distribution is one of a downstream increase in species richness by addition, but this concept is based on a small number of streams from the mid-west and southern United States, which are dominated by cyprinids. Further, the measure of species richness traditionally used, without including evenness, may not be providing an accurate reflection of the fish community. We hypothesize that in streams dominated by anadromous salmonids, fish community diversity will be affected by the presence of the anadromous species, and therefore be influenced by those factors affecting the salmonid population. Catamaran Brook, New Brunswick, Canada, provides a long-term data set to evaluate fish community diversity upstream and downstream of an obstruction (North American beaver Castor canadensis dam complex), which affects distribution of Atlantic salmon Salmo salar. The Shannon Weiner diversity index and community evenness were calculated for sample sites distributed throughout the brook and over 15 years. Fish community diversity was greatest upstream of the beaver dams and in the absence of Atlantic salmon. The salmon appear to depress the evenness of the community but do not affect species richness. The community upstream of the beaver dams changes due to replacement of slimy sculpin Cottus cognatus by salmon, rather than addition, when access is provided. Within Catamaran Brook, location of beaver dams and autumn streamflow interact to govern adult Atlantic salmon spawner distribution, which then dictates juvenile production and effects on fish community. These communities in an anadromous Atlantic salmon dominated stream do not follow the species richness gradient pattern shown in cyprinid-dominated streams and an alternative model for stream fish community distribution in streams dominated by anadromous salmonids is presented. This alternative model suggests that community distribution may be a function of semipermeable obstructions, streamflow and the distribution of the anadromous species affecting resident stream fish species richness, evenness, biomass and production.