Fractionation of δ15N and δ13C for Atlantic salmon and its intestinal cestode Eubothrium crassum

Stable isotopes of nitrogen (δ¹⁵N) and carbon (δ¹³C) were measured for Atlantic salmon Salmo salar and their intestinal cestode, Eubothrium crassum , sharing the same diet. Atlantic salmon muscle tissues were enriched in ¹⁵N and depleted in ¹³C compared to their prey (sprat Sprattus sprattus sprattus ) and their intestinal cestode. There was no significant difference in δ¹⁵N or δ¹³C between E. crassum and the sprat. Differences in nutrient uptake and intestine physiology between Atlantic salmon and E. crassum are discussed, as well as how these may give rise to different fractionations of stable isotopes between a host and its parasites. Furthermore, Atlantic salmon contained a significantly higher lipid content than their prey, which may partly explain differences in δ¹³C values between the host and its cestode. In addition, cestodes inhabiting lipid-rich hosts were also lipid rich. Larger Atlantic salmon were enriched in ¹⁵N compared to smaller fish. Cestodes inhabiting large hosts were also enriched in ¹⁵N compared to parasites living in smaller hosts. The last two results were explained by larger fish possibly feeding from a higher trophic level, or from larger and older prey, that resulted in both a higher lipid content and an enrichment in ¹⁵N.     

Macrotidal estuaries: a region of collision between migratory marine animals and tidal power development

Macrotidal estuaries of the inner Bay of Fundy are utilized by large numbers of migratory fishes, particularly dogfish, sturgeon, herring, shad, Atlantic salmon and striped bass as well as by other migratory marine animals, many of which have large body sizes (squid, Lamnid sharks, seals and whales). Tagging experiments indicate the fishes originate from stocks derived over the entire North American Atlantic coast from Florida to Labrador. Population estimates suggest up to 2.0 times 10⁶adult American shad (Alosa sapidissima) migrate through an individual embayment each year. These migrations are an integral part of the life history of the respective species and appear to be controlled in part by the near shore movements of ocean currents. In other regions of the world similar macrotidal estuaries exist (Cook Inlet, Alaska; Severn Estuary, U.K.) and they, like the Bay of Fundy, are linked in continuum to the local ocean currents. We propose that marine animals utilize all these regions in a manner similar to the Bay of Fundy estuaries and properly designed surveys will reveal their presence. Fish passage studies utilizing the Annapolis estuary low-head, tidal turbine on the Bay of Fundy have shown that turbine related mortality of 20–80% per passage occurs depending on fish species, fish size and the efficiency of turbine operation. We suggest that introduction of tidal turbines into open ocean current systems will cause widespread impact on marine populations resulting in significant declines in abundance.     

Preparation and Binding Properties of Radioiodinated Analogues of Dermorphin and Deltorphin with High Specificity for the μ- and δ-Opioid Receptors

Abstract: The synthesis, purification, chemical characterization, and binding properties of two ¹²⁵I-labeled analogues of dermorphin and deltorphin-I are described. Native deltorphin-I and [Lys⁷]dermorphin sequences were elongated by an aminopentyl chain on their C-terminal amide function and alkylated with the ¹²⁵I-labeled monoiodinated derivative of Bolton-Hunter reagent (BH^*). The resulting radiolabeled peptides, ε-BH^* [Lys⁷]dermorphin 5-aminopentylamide and ω-BH^* deltorphin-I 5-aminopentylamide, have kept most of the original properties of the parent peptides. They bind with high selectivity and specificity to the μ- (dermorphin analogue) or δ- (deltorphin-I analogue) opioid receptors from rat brain or from cells transfected with cDNAs encoding the μ and δ receptors. The autoradiographic distribution of specific binding sites for the ¹²⁵I-labeled dermorphin and deltorphin-I analogues in rat brain is in complete agreement with previously reported localizations of μ- and δ-opioid receptors. The two radiolabeled peptides are the best ligands of μ- and δ-opioid receptors currently available in terms of sensitivity, specificity, and selectivity.     

Key Residues Defining the μ-Opioid Receptor Binding Pocket: A Site-Directed Mutagenesis Study

Abstract: Structural elements of the rat μ-opioid receptor important in ligand receptor binding and selectivity were examined using a site-directed mutagenesis approach. Five single amino acid mutations were made, three that altered conserved residues in the μ, δ, and κ receptors (Asn¹⁵⁰ to Ala, His²⁹⁷ to Ala, and Tyr³²⁶ to Phe) and two designed to test for μ/δ selectivity (Ile¹⁹⁸ to Val and Val²⁰² to Ile). Mutation of His²⁹⁷ in transmembrane domain 6 (TM6) resulted in no detectable binding with [³H]DAMGO (³H-labeled d -Ala², N -Me-Phe⁴,Gly-ol⁵-enkephalin), [³H]bremazocine, or [³H]ethylketocyclazocine. Mutation of Asn¹⁵⁰ in TM3 produces a three- to 20-fold increase in affinity for the opioid agonists morphine, DAMGO, fentanyl, β-endorphin_1–31, JOM-13, deltorphin II, dynorphin_1–13, and U50,488, with no change in the binding of antagonists such as naloxone, naltrexone, naltrindole, and nor-binaltorphamine. In contrast, the Tyr³²⁶ mutation in TM7 resulted in a decreased affinity for a wide spectrum of μ, δ, and κ agonists and antagonists. Altering Val²⁰² to Ile in TM4 produced no change on ligand affinity, but Ile¹⁹⁸ to Val resulted in a four- to fivefold decreased affinity for the μ agonists morphine and DAMGO, with no change in the binding affinities of κ and δ ligands.     

Patterns of stable isotope signatures in willow warbler Phylloscopus trochilus feathers collected in Africa

We conducted stable isotope analyses of nitrogen and carbon on feathers obtained from willow warblers in Africa to find an explanation for a previously observed pattern of different δ¹⁵N and δ¹³C values across a migratory divide in central Scandinavia. A new data set confirms that north Scandinavian birds of the subspecies P. t. acredula have higher δ¹⁵N values than south Scandinavian birds of the subspecies P. t. trochilus . In Africa, we found significant differences for both δ¹⁵N and δ¹³C values among feathers collected from major geographical regions as well as between countries within regions. Isotope signatures of δ¹⁵N and δ¹³C in Scandinavian acredula matched well with those of willow warblers sampled in southern parts of Africa, but differed from samples obtained in East and West Africa. Isotope signatures in Scandinavian trochilus did not agree with the pattern in any of the three African regions (West, East or South). However, a more detailed analysis of the isotopic data in feathers from countries within West Africa, which is the wintering region of Swedish trochilus based on ringing recoveries, revealed a correspondence with samples from Liberia, the Ivory Coast and Nigeria.     

Regional differentiation of North American Atlantic salmon at allozyme loci

Allozyme variation was characterised by starch gel electrophoresis at 23 enzyme coding loci and one regulatory locus in Atlantic salmon from 53 rivers in Eastern Canada, encompassing the majority of the species' North American range. Variation among rivers was highly heterogeneous and eight of the 15 polymorphisms showed regionally restricted distributions. Nearest neighbour joining (NJ) analysis and multi‐dimensional scaling suggest six distinct regional groups; Labrador/Ungava, Gulf of Saint Lawrence, Newfoundland (excluding Gulf rivers), the Atlantic shore/Southern Uplands of Nova Scotia, the inner Bay of Fundy, and the outer Bay of Fundy. Approximately 25% of observed genetic variation was distributed among these regions with a weak though significant overall correlation of genetic and geographic distance (Mantel Test, r  = 0·255, P  = 0·005). Collectively, the rivers showed consistent divergence from European populations with strong bootstrap support for the two clusters across loci in the NJ analysis. Mean heterozygosity was 0·061 for both continental groups, but the European population showed more than twice the variation among populations. F _ST values were 0·076 and 0·176 for North America and Europe, respectively, with an overall F _ST of 0·330.     

Restricted matrilineal gene flow and regional differentiation among Atlantic salmon (Salmo salar L.) populations within the Bay of Fundy,eastern Canada

Matrilineal phylogenetic divergence among Atlantic salmon stocks of the Bay of Fundy in south eastern Canada is investigated. Sequence variation in two regions of the mitochondrial ND1 gene, encompassing 710 base pairs, is described for 168 salmon from 11 rivers. Mean overall haplotype and nucleotide diversity (h and pi) observed are 0.5014 and 0.00095, respectively. Nested clade analysis (NCA) and molecular analysis of variance (AMOVA) both point to highly restricted gene flow among rivers and show the haplotype distribution to be geographically structured. Variation among predefined regions of the Bay (16%) is greater than among populations within these regions (14%) The main regional differentiation occurs between rivers of the geographically isolated inner Minas Basin and those elsewhere in the Bay. Differentiation most probably reflects the pattern and nature of the historical processes associated with post-glacial colonisation of the area by salmon following the last Pleistocene glacial maximum c. 180,00 yrs BP.     

Kinetic characteristics of the sodium uptake mechanism during the development of embryos and fry of Atlantic salmon, Salmo salar L., in an improved water quality

The kinetics of active sodium uptake in dechorionated embryos, yolk-sac fry and start-feed fry of Atlantic salmon were compared in two groups reared either in low conductivity, untreated, river water (conductivity ∼ 46 μS cm⁻¹, pH 5.75), or in 'improved' river water buffered with sea water (conductivity ∼2200 μS cm ¹, pH 6.56), the latter treatment often being used in commercial hatcheries to avoid problems associated with periodic acidification.
Maximal transport rate ( V _max) increased during development in both groups but was always significantly higher in embryos and fry maintained in untreated river water. Values for K _m were not seen to vary during development up to 12 weeks after hatching and were not significantly different between groups, or from values reported for adult Atlantic salmon in fresh water.
The results are discussed with respect to the influence of Na⁺ concentrations in the perivitelline fluid of developing eggs and in the external medium surrounding fry on V _max and K _m. The ability of fry reared entirely in buffered river water to maintain sodium balance following transfer to untreated river water is also considered.     

Life-stage-dependent supergene haplotype frequencies and metapopulation neutral genetic patterns of Atlantic cod,Gadus morhua,from Canada's Northern cod stock region and adjacent areas

Among highly migratory fish species, nursery areas occupied by juveniles often differ from adult habitats. To better understand the spatial dynamics of Canada's Northern cod stock, juveniles caught off the east coast of Newfoundland and Labrador were compared to adults from the same region as well as individuals from other areas in Atlantic Canada using double-digest restriction site-associated DNA sequencing–derived single nucleotide polymorphisms. A reduced proportion of homozygotes with a chromosomal inversion located in linkage group 1 (LG1) was detected between juvenile and adult samples in the Northern cod stock region, potentially indicating age-dependent habitat use or ontogenetic selection for attributes associated with the many genes located in LG1. No selectively neutral genetic differences were found between samples from the Northern cod stock; nevertheless, significant differences were found between some of these samples and cod collected from St. Pierre Bank, Bay of Fundy, Browns Bank and the southern Scotian Shelf. Clustering analysis of variants at neutral loci provided evidence for three major genetic units: (a) the Newfoundland Atlantic Coast, (b) eastern and southern Gulf of St. Lawrence and Burgeo Bank and (c) the Bay of Fundy, Browns Bank and southern Scotian Shelf. Both adaptive and neutral population structure within the Northern cod stock should be considered by managers to promote demographic rebuilding of the stock.     

GEOGRAPHICAL VARIATION IN METRIC SKULL CHARACTERS AMONG PROPOSED SUBPOPULATIONS AND STOCKS OF HARBOR PORPOISE, PHOCOENA PHOCOENA, IN THE WESTERN NORTH ATLANTIC

A total of 433 skulls of harbor porpoises, Phocoena phocoena , were examined to evaluate geographical variation within the western North Atlantic, using analysis of covariance and discriminant analysis based on 45 measurements of skull components. The relationship between condylobasal length and body length was sigmoid in both sexes. Condylobasal length is generally less in adult males than in adult females.
Despite some overlap, morphological differences between different regions and even adjacent local stocks could be recognized by ANCOVA and discriminant analysis. The percentage of correct classification by discriminant functions was 65.2% for the four regional samples: (1) North Canadian Atlantic (which includes Gulf of St. Lawrence and Newfoundland), (2) Bay of Fundy, (3) Gulf of Maine-New York, and (4) Maryland-South Carolina. It was 65.9% for the four local geographical areas (10s of km) within the Bay of Fundy; Quoddy-region porpoises revealed a closer relationship with the southwestern Nova Scotia population than with other adjacent stocks. The results at this level imply that gene flow is restricted to some degree even among closely adjacent geographical units. Management of this species should aim, therefore, for conservation of a variety of local stocks.     

Stable isotope and pen feeding trial studies confirm the value of horseshoe crab Limulus polyphemus eggs to spring migrant shorebirds in Delaware Bay

We used stable isotope (SI) methods in combination with pen feeding trials to determine the importance of eggs of the Atlantic horseshoe crab Limulus polyphemus to migratory fattening of red knots Calidris canutus rufa and ruddy turnstones Arenaria interpres morinella during spring stopover in Delaware Bay. By manifesting measurable fractionation (ca +3‰) and rapid turnover, blood plasma δ¹⁵ nitrogen proved a functional marker for SI diet tracking during the short 3-week stopover. Blood samples from free-ranging knots (3 data sets) and turnstones (1 data set) produced similar convergence of plasma δ¹⁵N signatures with increasing body mass that indicated highly similar diets. Asymptotes deviated slightly (0.3‰ to 0.7‰) from that of captive shorebirds fed a diet of only crab eggs during stopover, thus confirming a strong crab egg-shorebird linkage. The plasma δ¹⁵N crab-egg diet asymptote was enriched ca +4.5‰ and therefore readily discriminated from that of either blue mussels Mytilus edulis or coquina clams Donax variabilis , the most likely alternative prey of knots in Delaware Bay. Crab eggs were highly palatable to captive knots and turnstones which achieved rates of mass gain (3–11 g/d) comparable to that of free-ranging birds. Peak consumption rates during hyperphagic events were 23,940 and 19,360 eggs/bird/d, respectively. The empirical conversions of eggs consumed to body mass gained (5,017 eggs/g for knots and 4,320 eggs/g for turnstones) indicate the large quantities of crab eggs required for the maintenance of these shorebird populations during stopover.     

Assessing seasonal changes in stock composition of Atlantic salmon catches in the Baltic Sea with genetic stock identification

The feasibility of using genetic stock identification to analyse seasonal changes in stock compositions of Atlantic salmon catches in the Baltic Sea was examined. The analysis employed seven variable allozyme loci from most of the potentially contributing stocks (16) from Finland and Sweden. Catch samples were collected from Finnish salmon fisheries in the eastern Bothnian Sea during the 1992 fishing season. Simulation studies were used to evaluate the feasibility of identifying Baltic salmon stocks with allozyme data. Special attention was paid to analysing the wild production of salmon stocks. Clear seasonal differences in stock composition were found. The estimates were compared with smolt production and Carlin-tag data. The proportions of the Neva and Oulujoki river stocks could be estimated as individual stocks, whereas the contributions of the remaining stocks were estimated as four composite stock groups. One of the groups consisted of wild stocks from the rivers Kalixälven and Simojoki. Identification of this group, which could be used as an index of wild production in the catches, requires catch sample sizes >300 salmon if <15% error is required.     

Normal ranges of some blood chemistry parameters in adult farmed Atlantic salmon, Salmo salar

Blood samples from healthy adult Atlantic salmon fed an optimal diet in net sea pens were collected at intervals from October to May. Haematological determinations and biochemical serum analyses were carried out on 20 fish in each of seven samples. The ranges of haemato-logical values for sample means were: haematocrit 44–49%, haemoglobin 8.9–10.4 g 100ml⁻¹, red blood cell count 0.85–1.10 × 10¹² l⁻¹, MCV 441–553 × 10⁻¹⁵ 1, MCH 94–106 × 10⁻⁶ g, MCHC 19.4–21.7 g 100ml⁻¹ and leucocrit 0.43–0.96%. The ranges of enzyme activities in serum, for sample means, were: alkaline phosphatase 647–988Ul⁻¹, aspartate aminotrans-ferase 202–351 Ul⁻¹ and alanine aminotransferase 4–8 Ul⁻¹. The ranges of the other parameters analyzed in serum were: total protein 41.6–56.6 gl⁻¹, albumin 18.3–24.3 gl⁻¹, albumin/total protein ratio 39.3–44.0%, creatinine 26–46 μmol, triglycerides 2.53–4.98 μmol and cholesterol 9.3–12.8 μmol. These values are considered to be the normal ranges in healthy fish. Variations due to seasonal changes, and the clinical significance of the selected parameters, are discussed. Data showing the reproducibility of the biochemical analyses in serum are presented.     

Uptake of salmon-derived nitrogen by mosses and liverworts in coastal British Columbia

Throughout forested watersheds bordering the North Pacific, anadromous salmon ( Oncorhynchus spp . ) are transferred into coastal forests by numerous predators and scavengers with remnants providing an important nutrient subsidy to riparian zones. The contribution of these nutrients to mosses and liverworts, which are the dominant ground cover of coastal forests, has not been investigated. We examine here δ¹⁵N isotope signatures and foliar percent N of eight moss and liverwort species, as well as moss and liverwort community structure, in multiple habitat blocks that vary in access to salmon nutrients from two watersheds on the central coast of British Columbia. For the most common moss Rhytidiadelphus loreus , we also examine δ¹⁵N values and foliar percent N among salmon carcass micro-sites and along wildlife trails. Overall, δ¹⁵N signatures ranged from 2‰ to 7‰ higher below the falls near the salmon stream than above the small falls that are impassable to salmon, or at the adjacent control watershed that had no salmon. Among micro-sites, δ¹⁵N and %N values were highest near bony carcass remnants from transfer during previous years and high adjacent to wildlife trails indicating spatial heterogeneity in the salmon nutrient pools of these forests. Species richness and prevalence of nitrogen-rich soil indicators were also highest in forests adjacent to the salmon stream. These data suggest an important contribution of salmon-derived nutrients to the non-vascular plants of the riparian zone of old-growth forests throughout the Pacific Rim.     

Factors affecting river entry of adult Atlantic salmon in a small river

In this study, effects of stock origin, fish size, water flow and temperature on time of river ascent of adult Atlantic salmon Salmo salar were tested. Brood stocks were collected in eight Norwegian rivers situated between 59 and 69° N. The fish were reared to smolts, individually tagged and released in the River Imsa, south-west Norway (59° N). Adults from all stocks approached the Norwegian coast concurrently, but Atlantic salmon ≥70 cm in natural tip length entered coastal water slightly earlier during summer than smaller fish. Atlantic salmon <70 cm, however, ascended the river significantly earlier and at lower water flow and higher water temperature than larger fish. Although largest in size, the fish from the northern populations (62–69° N) ascended the River Imsa almost 1 month earlier than those from the south (59–60° N). They seemed less restricted by the environmental factors than the fish originating from the more southern rivers. There was no apparent trend among years in time of river ascent. Maximum ascent per day occurred at water discharges between 12·5 and 15 m³ s⁻¹ and at water temperatures between 10 and 12·5° C. There was a significant positive correlation between water flow and river ascent during the first part of the upstream run from July to September with best correlation for September, when multiple regression analysis indicated that water temperature had an additional positive effect. Stock origin, fish size and water discharge were important variables influencing the upstream migration of Atlantic salmon in small rivers.     

Outlier SNPs show more genetic structure between two Bay of Fundy metapopulations of Atlantic salmon than do neutral SNPs

Atlantic salmon of Eastern Canada were once of considerable importance to aboriginal, recreational, and commercial fisheries, yet many populations are now in decline, particularly those of the inner Bay of Fundy (iBoF), which were recently listed as endangered. We investigated whether nonneutral SNPs could be used to assign individual Atlantic salmon accurately to either the iBoF or the outer Bay of Fundy (oBoF) metapopulations because this has been difficult with existing neutral markers. We first searched for markers under diversifying selection by genotyping eight captively bred Bay of Fundy (BoF) populations for 320 SNP loci with the Sequenom MassARRAY? system and then analysed the data set with four different F(ST) outlier detection programs. Three outlier loci were identified by both BayesFST and BayeScan whereas seven outlier loci, including the three previously mentioned, were identified by both Fdist and Arlequin. A subset of 14 nonneutral SNPs was more accurate (85% accuracy) than a subset of 67 neutral SNPs (75% accuracy) at assigning individual salmon back to their metapopulation. We then chose a subset of nine outlier SNP markers and used them to inexpensively genotype archived DNA samples from seven wild BoF populations using Invader? chemistry. Hierarchical AMOVA of these independent wild samples corroborated our previous findings of significant genetic differentiation between iBoF and oBoF salmon metapopulations. Our research shows that identifying and using outlier loci is an important step towards achieving the goal of consistently and accurately distinguishing iBoF from oBoF Atlantic salmon, which will aid in their conservation.     

Sublethal effects of the pesticide Diazinon on olfactory function in mature male Atlantic salmon parr

Diazinon, an organophosphate pesticide, had a sublethal effect on the olfactory system of mature male Atlantic salmon parr. The olfactory responses of the parr to prostaglandin F_2a (PGF_2a) were studied after exposure of the epithelium to different concentrations of Diazinon in water. Electrophysiological recordings from the epithelium indicated that the responses to this prostaglandin were significantly reduced at nominal concentrations as low as 1.0μg l⁻¹ and the threshold of detection was reduced 10-fold at 2.0 μg 1⁻¹ . Mature male salmon parr exposed for a period of 120 h to Diazinon (nominal concentrations 0.3, 0.8, 1.7, 2.7, 5.6, 13, 28 and 45 μg 1 ⁻¹) also had significantly reduced levels of the reproductive steroids, 17,20β-dihydroxy-4-pregnen-3-one, testosterone and gonadotrophin II in the blood plasma after priming with ovulated female salmon urine. Both prostaglandin F_2a and ovulated female urine are known to have important roles in synchronizing reproductive physiology and behaviour in salmonids as well as other fish species. The results are therefore discussed in relation to the possible sublethal effects of Diazinon on reproduction in the Atlantic salmon and possible effects on populations of salmonids.     

Foreword

Allozyme variation was characterised by starch gel electrophoresis at 23 enzyme coding loci and one regulatory locus in Atlantic salmon from 53 rivers in Eastern Canada, encompassing the majority of the species' North American range. Variation among rivers was highly heterogeneous and eight of the 15 polymorphisms showed regionally restricted distributions. Nearest neighbour joining (NJ) analysis and multi‐dimensional scaling suggest six distinct regional groups; Labrador/Ungava, Gulf of Saint Lawrence, Newfoundland (excluding Gulf rivers), the Atlantic shore/Southern Uplands of Nova Scotia, the inner Bay of Fundy, and the outer Bay of Fundy. Approximately 25% of observed genetic variation was distributed among these regions with a weak though significant overall correlation of genetic and geographic distance (Mantel Test, r = 0·255, P = 0·005). Collectively, the rivers showed consistent divergence from European populations with strong bootstrap support for the two clusters across loci in the NJ analysis. Mean heterozygosity was 0·061 for both continental groups, but the European population showed more than twice the variation among populations. F_ST values were 0·076 and 0·176 for North America and Europe, respectively, with an overall F_ST of 0·330.