A Rainbow Trout Lectin with Multimeric Structure

A novel lectin has been identified in rainbow trout serum and plasma. The lectin binds to Sepharose (an agarose polymer) in a calcium-dependent manner. Glucose, N-acetyl-glucosamine, mannose, N-acetyl-mannosamine, l-fucose, maltose and α-methyl-mannoside are good inhibitors of this binding, whereas glucosamine and d-fucose inhibits to a lesser degree and mannosamine and galactose do not inhibit the binding to Sepharose. When analysed by SDS-PAGE under non-reducing conditions, the lectin appears as a characteristic ladder of bands with approximately 16 kDa between consecutive bands. Upon reduction, the lectin appears as a 16-kDa band. On size-exclusion chromatography of trout serum and plasma, the protein emerges over a broad range corresponding to sizes from about 2000 kDa to less than 200 kDa. The NH₂-terminal sequence (AAENRNQXPPG) shows no significant homology with known proteins. Because of the characteristic appearance in non-reducing SDS-PAGE and the lectin activity, we propose to name the protein “ladderlectin.”     

The Effect of Ration on Acclimation to Environmental Acidity in Rainbow Trout

Freshwater salmonids exposed to low environmental pH typically suffer a net loss of ions, primarily Na⁺ and Cl^–, across the gills, resulting in reduced plasma and tissue ion concentrations. However, in recent experiments in our laboratory, juvenile rainbow trout, Oncorhynchus mykiss, fed a ration of 1% body weight d^–1 or greater showed no ionoregulatory disturbance during chronic, sublethal acidification. This raised the possibility that these fish had acclimated to low pH in that they would be better able to withstand further, more severe acidification than fish that had no prior experience of acid conditions: previous studies had concluded that such acclimation does not occur. This hypothesis was tested by measuring unidirectional ion fluxes during a 24h acute acid challenge (pH 4.2) in juvenile rainbow trout that had previously been exposed to either ambient pH 6.2 (naive fish) or sublethal low pH 5.2 (acid pre-exposed fish) for 90 days, and fed a ration of either 1.0 or 0.25% d^–1 (wet basis). No mortalities were observed during the acute acid challenge in the fish fed the higher ration and no differences between the two groups in the response of Na⁺ fluxes were observed. Sodium influx in both groups was significantly inhibited throughout the challenge and Na⁺ net flux was significantly stimulated over the first 6h. Prior to the acute acid challenge, the fish fed the lower ration that had previously been exposed to pH 5.2 had significantly lower plasma ion concentrations than those fish previously exposed to pH 6.2. Both groups suffered mortalities; those of the naive fish (22% by 24h) being markedly lower than those of the acid pre-exposed fish (68% by 24h). However, there were no significant differences in either Na⁺ or Cl^– fluxes between the two groups of fish during the acid challenge: both showed significant inhibition of ion influxes and significantly greater net ion losses, resulting in reduced plasma ion concentrations. These results indicate that rainbow trout are unable to acclimate to environmental acidification irrespective of the availability of dietary salts.     

Variability of kokanee and rainbow trout food habits,distribution, and population dynamics,in an ultraoligotrophic lake with no manipulative management

Crater Lake is a unique environment to evaluate the ecology of introduced kokanee and rainbow trout because of its otherwise pristine state, low productivity, absence of manipulative management, and lack of lotic systems for fish spawning. Between 1986 and 2004, kokanee displayed a great deal of variation in population demographics with a pattern that reoccurred in about 10 years. We believe that the reoccurring pattern resulted from density dependent growth, and associated changes in reproduction and abundance, driven by prey resource limitation that resulted from low lake productivity exacerbated by prey consumption when kokanee were abundant. Kokanee fed primarily on small-bodied prey from the mid-water column; whereas rainbow trout fed on large-bodied prey from the benthos and lake surface. Cladoceran zooplankton abundance may be regulated by kokanee. And kokanee growth and reproductive success may be influenced by the availability of Daphnia pulicaria, which was absent in zooplankton samples collected annually from 1990 to 1995, and after 1999. Distribution and diel migration of kokanee varied over the duration of the study and appeared to be most closely associated with prey availability, maximization of bioenergetic efficiency, and fish density. Rainbow trout were less abundant than were kokanee and exhibited less variation in population demographics, distribution, and food habits. There is some evidence that the population dynamics of rainbow trout were in-part related to the availability of kokanee as prey.     

Food habits of Coexisting Salmonines above and below Stronach Dam in the Pine River, Michigan

Although dam removal has become an increasingly popular tool for river restoration, there is limited knowledge regarding the ecological effects of dam removal. The purpose of our study was to document feeding habits of coexisting brook charr, brown trout, and rainbow trout above and below a dam that is in the process of a staged removal. Modification of sediment transport caused by Stronach Dam since 1912 has affected stream channel configuration, fish habitat, and many other physical and biological processes. In order to document salmonine feeding habits above and below the dam, we selected zones to represent downstream conditions and areas of river upstream of the dam that encompassed the original reservoir and a stretch of river further upstream that was not hydraulically influenced by the dam. Because physical habitat largely governs aquatic community composition in streams, we expected these effects to be reflected in the fish and macroinvertebrate communities. In particular, we expected limited prey availability and salmonine feeding in the impacted upstream and downstream zones characterized by fine substrate composition and greater macroinvertebrate diversity and salmonine feeding opportunities in the non-impacted zone characterized by coarse substrate. We also expected mean percent wet stomach content weights to be higher downstream, as other studies have documented an increase in piscivory on blocked migratory prey species downstream of dams. Contrary to expectations, the downstream zone of the river contained the highest abundance of drifting invertebrate taxa and, although differences in habitat occurred among the zones, the diversity of drifting macroinvertebrates and stomach contents of salmonines were similar throughout the river. Thus, in this case, the presence of altered habitat caused by a dam did not appear to negatively affect salmonine food habits. Consequently, we expect no major changes in salmonine food habits after the dam removal is completed.     

Evidence of partial anadromy and resident-form dispersal bias on a fine scale in populations of Oncorhynchus mykiss

We examine sympatric anadromous (steelhead) and nonanadromous (resident) rainbow trout (Oncorhynchus mykiss) from neighboring locations to test three hypotheses: (1) the sympatric life history types are not genetically different; (2) fine-scale dispersal is the same for both sexes, and (3) fine-scale dispersal is the same for steelhead and resident individuals. Data from 13 microsatellite loci reveal no genetic difference between sympatric steelhead and resident O. mykiss but moderate population structure (F _ST=0.019–0.028) between adjacent samples, regardless of life history type. Our results provide further evidence of partial anadromy and suggest that geographic proximity and genetic history, more than migratory type, should be considered when identifying populations for use in restoration of local genetic diversity. We find evidence of resident-form dispersal bias on a fine spatial scale, however, we find no evidence that fine-scale dispersal varies by gender. Conservation strategies should aim to maintain resident and anadromous forms when they occur in sympatry, as they may be important in facilitating gene flow on small and large spatial scales, respectively.     

Diet, habitat use, and relative abundance of pampas fox (Pseudalopex gymnocercus) in northern Patagonia, Argentina

Diet, habitat use and relative abundance of pampas fox Pseudalopex gymnocercus were studied in grassland, sand dune, and scrubland habitats, on the Bahía San Blas and Isla Gama Reserve, Argentina. Scat contents showed a generalist diet, where mammals, insects, and fruits were the main food items. Dietary composition differed among habitats; fruits were mainly consumed in sand dunes; insects, mammals, and fruits predominated in grassland; while insects and mammals were frequent in scrubland habitat. The European hare was the most common vertebrate prey and had the major contribution to the biomass of consumed prey. Pampas fox was the most common carnivore of the reserve and used all available habitats, frequenting more regularly scrubland and sand dune than grassland habitat.

Zusammenfassung

Nahrung, Habitatnutzung und relative Abundanz von Pampasfüchsen (Pseudalopex gymnocercus) im nördlichen Patagonien, Argentinien Die Nahrung, Habitatnutzung und relative Abundanz von Pampasfüchsen Pseudalopex gymnocercus wurde auf Grasflächen, Sanddünen und in Strauchhabitaten des Bahía San Blas and Isla Gama Reserve, Argentinien untersucht. Die Analyse von Kotproben ergab eine typische Diät für Generalisten, wobei Säugetiere, Insekten und Früchte die wichtigsten Nahrungsbestandteile waren. Die Nahrungszusammensetzung unterschied sich zwischen verschiedenen Lebensräumen. Früchte wurden hauptsächlich in Sanddünen konsumiert, Insekten, Säugetiere und Früchte auf Grasflächen; und Insekten und Säugetiere in Strauchhabitaten. Der europäische Hase war die häufigste Beuteart unter den Wirbeltieren und stellte den Hauptanteil an der Biomasse genutzter Beute. Der Pampasfuchs ist das häufigste Raubtier im Reservat und nutzt alle vorhandenen Habitate, jedoch kommt er etwas häufiger in Strauchhabitaten und Sanddünen vor als auf Grasflächen.     

A cDNA microarray assessment of gene expression in the liver of rainbow trout (Oncorhynchus mykiss) in response to a handling and confinement stressor

A purpose-designed microarray platform (Stressgenes, Phase 1) was utilised to investigate the changes in gene expression within the liver of rainbow trout during exposure to a prolonged period of confinement. Tissue and blood samples were collected from trout at intervals up to 648 h after transfer to a standardised confinement stressor, together with matched samples from undisturbed control fish. Plasma ACTH, cortisol, glucose and lactate were analysed to confirm that the neuroendocrine response to confinement was consistent with previous findings and to provide a phenotypic context to assist interpretation of gene expression data. Liver samples for suppression subtractive hybridisation (SSH) library construction were selected from within the experimental groups comprising “early” stress (2–48 h) and “late” stress (96–504 h). In order to reduce redundancy within the four SSH libraries and yield a higher number of unique clones an additional subtraction was carried out. After printing of the arrays a series of 55 hybridisations were executed to cover 6 time points. At 2 h, 6 h, 24 h, 168 h and 504 h 5 individual confined fish and 5 individual control fish were used with control fish only at 0 h. A preliminary list of 314 clones considered differentially regulated over the complete time course was generated by a combination of data analysis approaches and the most significant gene expression changes were found to occur during the 24 h to 168 h time period with a general approach to control levels by 504 h. Few changes in expression were apparent over the first 6 h. The list of genes whose expression was significantly altered comprised predominantly genes belonging to the biological process category (response to stimulus) and one cellular component category (extracellular region) and were dominated by so-called acute phase proteins. Analysis of the gene expression profile in liver tissue during confinement revealed a number of significant clusters. The major patterns comprised genes that were up-regulated at 24 h and beyond, the primary examples being haptoglobin, β-fibrinogen and EST10729. Two representative genes from each of the six k-means clusters were validated by qPCR. Correlations between microarray and qPCR expression patterns were significant for most of the genes tested. qPCR analysis revealed that haptoglobin expression was up-regulated approximately 8-fold at 24 h and over 13-fold by 168 h.     

Variation of vertebral number in juvenile Oncorhynchus mykiss in relation to upstream distance from the ocean

Vertebral counts in fishes are set early in development by a complex interaction of genetic and environmental factors. Recently, R. M. McDowall showed that vertebral counts can differ between diadromous and non-diadromous galaxiids, and suggested similar life history-based variation might also occur in the closely related Salmonidae. We examined variation in vertebral counts of juvenile rainbow trout, Oncorhynchus mykiss, from within a single large basin where anadromous and non-anadromous forms co-occur. Juveniles were collected at 29 sites within the Klamath River basin using a nested sampling design. Mean vertebral counts varied among regions, among complexes within regions, and among sites within complexes. Regression showed that mean vertebral counts decreased with increasing distance from the ocean. This decrease could not be explained by differences in temperature within the basin during early development. However, the decrease is consistent with the expectation that non-anadromous forms have fewer vertebrae and are more common at sites distant from the ocean, and suggests that further study of the relationship between vertebral counts and life history form in O. mykiss is warranted.

Fatty acid composition of Oncorhynchus mykiss during embryogenesis and other developmental stages

Fatty acid compositions of rainbow trout Oncorhynchus mykiss (Walbaum, 1792) was determined during embryogenesis, yolk-sac fry, swim up fry and fry stages. Embryonic and yolk-sac development of fertilized eggs were followed until fry weighed 25 g and samples were taken from each stage to analyse fatty acid composition. Analyses of fatty acid composition were performed by gas chromatography mass spectrometry. Throughout all the developmental stages of O. mykiss important changes, such as an increase in the percentage of 18:2, 18:3 and 20:5 and as a decrease in the percentage of 14:0, 16:2, 20:0, 20:2, 20:3, 20:4 and 22:0 were observed in the amounts of fatty acid of developing eggs. During transition from the embryonic to the yolk-sac stage, an important increase in 20:0, 20:3, 20:5, 22:0 and a decrease in 18:3, 22:1 was determined between 45-day old embryo and yolk-sac fry. During transition from the 43 day yolk-sac stage to swim up stage, a decrease in the percentage of 18:0, 20:2 and an increase in the percentage of 14:0, 16:2, 18:2, 18:3 was seen. In all the observed stages, no qualitative but significant quantitative variations were determined in the fatty acid composition of O. mykiss. It may be concluded that 14:0, 16:0, 16:1, 18:0, 18:1 18, 20 and 22 polyunsaturated fatty acids are necessary for normal development of O. mykiss during their life stages.     

Rainbow trout (Oncorhynchus mykiss) sIgM- leucocytes secrete an interleukin-2 like growth factor after mitogenic stimulation in vitro

Two secreted proteins were detected in culture supernatants of PHA or PMA stimulated, immunomagnetically separated, sIgM(-) leucocytes of rainbow trout (Oncorhynchus mykiss) with 60kDa and with 12-15kDa (multiple bands). So called conditioned media (CM), containing these proteins, induced significant activation of blood and head kidney leucocytes. Immunomagnetically separated, naive as well as PHA activated sIgM(-) T lymphocytes and LPS prestimulated sIgM(+) B lymphocytes could be identified to be responding to these secreted proteins. Using a monoclonal antibody specific for mouse IL-2 (clone JES6-1A12), one of the multiple 12-15kDa proteins could be stained in Western blots. It was also shown that the induced proliferation was due to this protein in the CM, as the same anti-IL-2 mab was able to block the CM induced proliferation. Furthermore, survival of the IL-2 dependent mouse cell line HT-2 was enhanced after addition of various concentrations of CM. The data presented show, for the first time, that mitogen stimulated trout sIgM(-) leucocytes secrete a cytokine like growth factor sharing functional and structural similarities with mammalian IL-2.     

Effect of acute and chronic epinephrine administration on clearance and metabolism of [3H]-epinephrine in trout (Oncorhynchus mykiss)

Several studies have measured the rate of catecholamine clearance, metabolism, and tissue accumulation in fish. However, no information is available on the effect of repeated stress or high circulating catecholamine levels on catecholamine clearance and metabolism. We measured the clearance and metabolism of [³H]-epinephrine (approximately 0.1 g·kg^-1) in SW-acclimated rainbow trout subjected to acute (five injections in 1 day) and chronic (4 days; five injections per day) administration of 4.0 g·kg^-1 epinephrine or saline. In addition, a saturation experiment, where 4.0 g·kg^-1 of unlabelled epinephrine was injected concurrently with [³H]-epinephrine, investigated whether catecholamine clearance and metabolism are affected by high circulatin levels. Neither the rate constants for catecholamine clearance, nor the post-injection proportions of unmetabolised [³H]-epinephrine, deaminated [³H]-epinephrine and O-methylated [³H]-epinephrine were affected by the acute or chronic injection protocols. The concurrent injection of [³H]-epinephrine and 4.0 g·kg^-1 of unlabelled epinephrine resulted in an elevated postinjection ³H:¹⁴C ratio, but increased proportions of O-methylated [³H]-epinephrine and reduced proportions of unmetabolised [³H]-epinephrine. We conclude that in fish (1) catecholamine clearance and metabolism are unlikely to be compromised by repeated exposure to acute stressors; (2) catecholamine extraction and/or metabolism is enhanced when circulating levels are high; and (3) there is a marked capacity to rapidly (minutes) clear and inactivate catecholamines that are released in response to stressful stimuli.Abbreviations CA catecholamines - dpm disintegrations per minute - MAO monoamine oxidase - COMT catecholamine-O-methyltransferase - MOPEG 3-methoxy-4-hydroxyphenylglycol - VMA 3-methoxy-4-hydroxymendelic acid - SW seawater - HPLC high-performance liquid chromatography     

Isolation and characterization of the receptor for vitellogenin from follicles of the rainbow trout,Oncorhynchus mykiss

The isolation and characterization of the receptor for vitellogenin from follicle membranes of the rainbow trout, Oncorhynchus mykiss, is described. Follicle membrane proteins subjected to SDS-polyacrylamide gel electrophoresis and subsequently to either protein staining or ligand blotting with radiolabelled vitellogenin (¹²⁵iodine-vitellogenin) demonstrated that the vitellogenin receptor has an apparent molecular mass of 200 kD (probably comprising of two 100-kD subunits) under non-reducing conditions. The vitellogenin binding sites were identified as specific receptors: binding was saturable and the binding sites were both tissue specific to follicle membranes and exhibited ligand specificity. Scatchard analyses of specific binding data revealed a single class of binding sites with a high affinity for rainbow trout vitellogenin (K _d=8.2·10^-9 mol·1^-1). Both brown trout, Salmo trutta, vitellogenin and carp, Cyprinus carpio, vitellogenin were able to displace the radiolabelled rainbow trout vitellogenin from its receptor, although they were less effective than rainbow trout vitellogenin.Abbreviations B _max maximum number of binding sites available - BSA bovine serum albumin - bt-VTG brown trout vitellogenin - c-VTG earp vitellogenin - HEPES N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid - K _d dissociatian constant - NCM nitrocellulose membranes - PMSF phenylmethylsulphonylfluoride - rt-VTG rainbow trout vitellogenin - VTG vitellogenin     

Effects of an Introduced Piscivore on Native Trout: Insights from a Demographic Model

Introductions of exotic species pose a significant threat to the persistence of many native populations, including many inland fishes. In 1994, piscivorous lake trout (Salvelinus namaycush) were discovered in Yellowstone Lake, Yellowstone National Park, Wyoming, USA, one of the last strongholds of the native Yellowstone cutthroat trout (Oncorhynchus clarki bouvieri). Predation by lake trout is expected to lead to a substantial decline in the native cutthroat trout population, which may have significant negative consequences for terrestrial predators that depend on cutthroat trout for prey and for the recreational fishery of the Park. We developed a matrix demographic model for the cutthroat trout population in Yellowstone Lake to identify the life stages that are most critical for understanding population dynamics. Parameter estimates (vital rates) were manipulated to explore the possible consequences of lake trout invasion. Comparisons of our results with current estimates of population trend and age structure suggested that our model reflected current conditions of the system. Elasticity analysis of the model revealed that population growth was most sensitive to annual survival of young trout, the group that is expected to be most vulnerable to lake trout predation. Projection of our deterministic model suggested that, in addition to a decline in abundance of cutthroat trout, the effects of lake trout may be manifest as changes in age and breeding structure of the population. Simulations of a stochastic version of the model indicated that a 60% or greater decline in the cutthroat trout population could be expected within 100 years if the lake trout population were permitted to grow uncontrolled. However, an effective control strategy that prevented the establishment of a large population of lake trout substantially reduced population decline, although the reduction in the availability of adult trout to terrestrial predators and anglers may be still be substantial (20–40%). In addition to current control activities in place in the Park, we recommend a renewed emphasis on understanding and monitoring juvenile life stages of cutthroat trout. Our results demonstrate the value of existing data sets for developing models to estimate the potential impact of biological invasions on the management and conservation of native populations, especially when opportunities and resources for additional empirical studies are limited.     

Invading rainbow trout usurp a terrestrial prey subsidy from native charr and reduce their growth and abundance

Movements of prey organisms across ecosystem boundaries often subsidize consumer populations in adjacent habitats. Human disturbances such as habitat degradation or non-native species invasions may alter the characteristics or fate of these prey subsidies, but few studies have measured the direct effects of this disruption on the growth and local abundance of predators in recipient habitats. Here we present evidence, obtained from a combined experimental and comparative study in northern Japan, that an invading stream fish usurped the flux of allochthonous prey to a native fish, consequently altering the diet and reducing the growth and abundance of the native species. A large-scale field experiment showed that excluding terrestrial invertebrates that fell into the stream with a mesh greenhouse reduced terrestrial prey in diets of native Dolly Varden charr (Salvelinus malma) by 46–70%, and reduced their growth by 25% over six weeks. However, when nonnative rainbow trout (Oncorhynchus mykiss) were introduced, they monopolized these prey and caused an even greater reduction of terrestrial prey in charr diets of 82–93%, and reduced charr growth by 31% over the same period. Adding both greenhouse and rainbow trout treatments together produced similar results to adding either alone. Results from a comparative field study of six other stream sites in the region corroborated the experimental findings, showing that at invaded sites rainbow trout usurped the terrestrial prey subsidy, causing a more than 75% decrease in the biomass of terrestrial invertebrates in Dolly Varden diets and forcing them to shift their foraging to insects on the stream bottom. Moreover, at sites with even low densities of rainbow trout, biomass of Dolly Varden was more than 75% lower than at sites without rainbow trout. Disruption of resource fluxes between habitats may be a common, but unidentified, consequence of invasions, and an additional mechanism contributing to the loss of native species Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Geographic patterns of introgressive hybridization between native Yellowstone cutthroat trout (<Emphasis Type="Italic">Oncorhynchus clarkii bouvieri)</Emphasis> and introduced rainbow trout (<Emphasis Type="Italic">O. mykiss</Emphasis>) in the South Fork of the Snake River watershed,Idaho

Throughout its native range, the Yellowstone cutthroat trout (YCT), Oncorhynchus clarkii bouvieri, is declining dramatically in both abundance and distribution as a result of introgression with introduced rainbow trout (RBT), O. mykiss. We sampled over 1,200 trout from the South Fork of the Snake River (SFSR) watershed, in southeastern Idaho and western Wyoming, and measured the extent of introgression of RBT genes into native gene pools of YCT using seven species-specific, co-dominant nuclear genetic markers. We also used mitochondrial DNA (mtDNA) haplotype differences between the two parental trout species to determine the directionality of the hybridization. We found low levels of RBT introgression (only 7% of sampled individuals had one or more RBT alleles) into YCT gene pools, with the majority of hybrids (78%) occurring in mainstem localities of the SFSR and in lower elevation reaches of certain tributaries. Hybridization was bidirectional with respect to mtDNA haplotype, but the majority of hybrids (75%) had YCT maternal haplotypes, indicative of the greater proportion (90%) of YCT-genotypes in the SFSR watershed. The primary factor influencing the geographic distribution of RBT introgressed individuals was fluvial distance from localities of stocking origin. To a lesser extent, elevation, also influenced the distribution of hybrid genotypes, with several entire tributaries and all upper elevation reaches within tributaries harboring only YCT-genotypes. Important management implications of the study suggest targeting particular tributaries and upper reaches within tributaries for YCT protection and exclusion of RBT hybrid colonization.     

Diel Habitat Partitioning by Bull Charr and Cutthroat Trout During Fall and Winter in Rocky Mountain Streams

We used underwater observation to determine diel habitat partitioning between bull charr, Salvelinus confluentus, and cutthroat trout, Oncorhynchus clarki, during fall and winter (0.1–8.3°C) in two Rocky Mountain streams that differed in habitat availability. The majority (>70%) of both species emerged from concealment cover at night, though bull charr exhibited a greater tendency for nocturnal behavior than cutthroat trout. Differences in day and night counts were most pronounced at temperatures <3°C, when very few fish of either species were observed in the water column during the day, but both species were common at night. Both species used concealment cover of large woody debris and boulder substrate crevices in deep pools during the day. At night, fish emerged from cover and habitat use shifted to shallow water with low cover. Microhabitat partitioning among species and size classes occurred at night, cutthroat trout moving into shallower, faster water that was farther from cover compared to bull charr. Smaller fish of both species occupied focal positions in slower, shallower water closer to the substrate than larger fish. Large, mixed-species aggregations also were common in beaver ponds both day and night. High variation in diel and site-specific winter habitat use suggests the need for caution in developing habitat suitability criteria for salmonids based solely on daytime observations or on observations from a few sites. Our results support the need to incorporate nocturnal habitat use and partitioning in studies of salmonid ecology.