Arctic charr brain transcriptome strongly affected by summer seasonal growth but only subtly by feed deprivation.

BACKGROUND: The Arctic charr (Salvelinus alpinus) has a highly seasonal feeding cycle that comprises long periods of voluntary fasting and a short but intense feeding period during summer. Therefore, the charr represents an interesting species for studying appetite-regulating mechanisms in fish. RESULTS: In this study, we compared the brain transcriptomes of fed and feed deprived charr over a 4 weeks trial during their summer feeding season. Despite prominent differences in body condition between fed and feed deprived charr at the end of the trial, feed deprivation affected the brain transcriptome only slightly. In contrast, the transcriptome differed markedly over time in both fed and feed deprived charr, indicating strong shifts in basic cell metabolic processes possibly due to season, growth, temperature, or combinations thereof. The GO enrichment analysis revealed that many biological processes appeared to change in the same direction in both fed and feed deprived fish. In the feed deprived charr processes linked to oxygen transport and apoptosis were down- and up-regulated, respectively. Known genes encoding for appetite regulators did not respond to feed deprivation. Gene expression of Deiodinase 2 (DIO2), an enzyme implicated in the regulation of seasonal processes in mammals, was lower in response to season and feed deprivation. We further found a higher expression of VGF (non-acronymic) in the feed deprived than in the fed fish. This gene encodes for a neuropeptide associated with the control of energy metabolism in mammals, and has not been studied in relation to regulation of appetite and energy homeostasis in fish. CONCLUSIONS: In the Arctic charr, external and endogenous seasonal factors for example the increase in temperature and their circannual growth cycle, respectively, evoke much stronger responses in the brain than 4 weeks feed deprivation. The absence of a central hunger response in feed deprived charr give support for a strong resilience to the lack of food in this high Arctic species. DIO2 and VGF may play a role in the regulation of energy homeostasis and need to be further studied in seasonal fish.

c-myc in whitefish (Coregonus lavaretus): structure, expression, and insights into possible posttranscriptional regulatory mechanism.

c-myc has a crucial function in growth control, differentiation, and apoptosis of vertebrate cells. Despite the important role of c-myc in mediating the biological effects, studies of c-myc gene expression and factors that control it in organisms other than mammals, such as fish, have been rare. In the current study, we asked whether c-myc mRNA of whitefish, a feasible organism for pollution monitoring in aquatic systems and a model in toxicological research, contains activity sites for regulatory motifs in its 5'- and 3'-UTRs, similar to those found in mammals. We were particularly interested in whether miRNA-34, a known negative regulator of c-myc's in mammals, is able to regulate c-myc in fish. To answer these questions, we determined the mRNA sequence of whitefish c-myc and inferred the structure of the protein that it codes for. We found that the active sites of mRNA and structures of the inferred c-myc protein are similar to those found in mammals and other fish. Remarkably, levels of c-myc mRNA expression were very high in ovaries compared to other tissues of whitefish, thus corroborating previous data in fish. Using bioinformatic searches on c-myc 3'-UTR, we confirmed the presence of two miRNA-34a (miR-34a) response elements. Luciferase reporter assay showed that activity of reporters containing either the miR response elements or entire c-myc 3'-UTR was significantly reduced (p < 0.001) by ectopic expression of miR-34a. Therefore, we further investigated possible involvement of miR-34a in c-myc gene silencing by profiling the expression of both genes in livers of whitefish treated for 8, 24, 48 h with MC-LR, a potent c-myc inducer in mammals. Although the difference was only significant at p = 0.08, the expression of c-myc mRNA in challenged whitefish after 24 h of the treatment was notably higher than that in livers of control fish. Concurrently, we noticed slight but significant up-regulation of miR-34a after 24 and 48 h of the challenge (p < 0.05); however, we found no significant correlation of the c-myc mRNA levels and miR-34a expression. Together, these results suggest that miR-34a might regulate c-myc gene expression in whitefish liver; however, their involvement in MC-LR hepatotoxicity should be clarified in future studies.

Inter- and intra-specific patterns of density dependence and population size variability in Salmoniformes.

Population dynamics are typically affected by a combination of density-independent and density-dependent factors, the latter of which have been conceptually and theoretically linked with how variable population sizes are over time-which in turn has been tied to how prone populations are to extinction. To address evidence for the occurrence of density dependence and its relationship with population size variability (pv), we quantified each of these for 126 populations of 8 species of Salmoniformes. Using random-effects models, we partitioned variation in the strength of density dependence and the magnitude of pv between and within species and estimated the correlation of density dependence and population size variability at both the between- and within-species levels. We found that variation in the strength of density dependence was predominately within species (I(2) = 0.12 [corrected] variation in population size variability was distributed both between and within species (I(2) = 0.40). Contrary to theoretical and conceptual expectations, the strength of density dependence and the magnitude of population size variability were positively correlated at the between species level (r = 0.90), although this estimate had 95 % credibility intervals (Bayesian analogues to confidence intervals) that overlapped zero. The within-species correlation between density dependence and population size variability was not distinguishable from zero. Given that density dependence for Salmoniformes was highly variable within species, we next determined the joint effects of intrinsic (density-dependent) and extrinsic (density-independent) factors on the population dynamics of a threatened salmonid, the Lahontan cutthroat trout (Oncorhynchus clarkii henshawi). We found that density-dependent and -independent factors additively contributed to population dynamics. This finding suggests that the observed within-species variability in density dependence might be attributable to local differences in the strength of density-independent factors.

[Ultrastructure of granulocytes of bony fishes (orders Salmoniformes, Cypriniformes, Perciformes)].

Analysis of data on utrastructure of granulocytes of freshwater and marine bony fish of orders Salmoniformes, Cypriniformes, and Perciformes showed that in all studied species there were revealed two types of granulocytes - neutrophils and eosinophils. The exception was the bluefish Pomatomus saltatrix L. whose pronephros hemopoietic tissue was found to contain one type of the granulocytic line - neutrophils. The identification parameters of granular leukocytes are specific granules filling the cytoplasm. The main form of specific granules in neutrophils of bony fish of various phylogenetic groups is an elongated granule with different distribution of fibrils or a granule that has crystalloid formed from fibrils. The main form of eosinophil granules - large, electron-dense, homogenous.

Historical and contemporary trophic niche partitioning among Laurentian Great Lakes coregonines.

Anthropogenic activities have significantly altered freshwater fish communities. Extirpations of deepwater coregonines (Coregonus spp.), a diverse group of fish species, have left vast areas of the Laurentian Great Lakes devoid of a deepwater fish community. Currently, fisheries managers are considering restoring populations by reintroducing deepwater coregonines from Lake Superior and Lake Nipigon. However, little is known about the historical ecology of deepwater coregonines, and species characterization has proved difficult. We used stable isotope analysis of museum-preserved and contemporary specimens to investigate if (1) coregonine species historically occupied distinct niches and (2) the pattern of trophic niche partitioning has changed over the last century. Across all lakes, individual species occupied distinct trophic niches, confirming that these species were ecologically distinct. Understanding trophic niche partitioning helps resolve uncertainty about distinctness of species within and across lakes and may provide a better ecological basis for rehabilitation of Great Lakes food webs and ecosystems.

Genetic analysis of threatened Australian grayling Prototroctes maraena suggests recruitment to coastal rivers from an unstructured marine larval source population.

Population genetic variation of Australian grayling Prototroctes maraena was examined to determine whether the dispersal strategy of this amphidromous species favours retention of larvae and juveniles in close proximity to their natal river, or mixing of populations via marine dispersal. Variation in microsatellite and mitochondrial DNA markers was unstructured and differentiation was indistinguishable from zero across four coastal rivers spanning approximately one-quarter of the continental range of the species. This result indicates that the marine larval and juvenile phase probably facilitates extensive gene flow among coastal rivers and agrees with a previous analysis of otolith chemistry that suggested larvae probably move into the sea rather than remain in estuaries. It appears likely that the dispersal strategy of P. maraena would enable recolonization of rivers that experience localized extinction provided that connectivity between freshwater habitats and the sea is sufficient to permit migration and that enough source populations remain intact to support viability of the wider population.

Phylogenetic position of the enigmatic Lepidogalaxias salamandroides with comment on the orders of lower euteleostean fishes.

This study examines phylogenetic placement of the enigmatic Western Australian Lepidogalaxias, and extends previous studies by including eight new taxa to enable re-examination phylogenetic relationships of lower euteleostean fishes at the ordinal level, based on mitochondrial genomes from 39 ingroup taxa and 17 outgroups. Our results suggest that Lepidogalaxias occupies a basal position among all euteleosts, in contrast with earlier hypotheses that variously suggested a closer relationship to esocid fishes, or to the galaxiid Lovettia. In addition our evidence shows that Osmeriformes should be restricted Retropinnidae, Osmeridae, Plecoglossidae and Salangidae. This reduced Osmeriformes is supported in our results as the sister group of Stomiiformes. Galaxiidae, which is often closely linked to Osmeriformes, emerges as sister group of a combined Osmeriformes, Stomiiformes, Salmoniformes, Esociformes and Argentiformes, and we give Galaxiiformes the rank of order to include all remaining galaxioid fishes (Galaxias and allied taxa, Aplochiton and Lovettia). Our results also support a sister group relationship between Salmoniformes and Esociformes, which are together the sister group of Argentiniformes.

Effects of food deprivation on refuge use and dispersal in juvenile North Sea houting Coregonus oxyrinchus under experimental conditions.

This study tested the influence of energetic state on refuge use and dispersal in juvenile North Sea houting Coregonus oxyrinchus in an artificial stream. Food-deprived fish spent more time outside refuges than well-fed fish; however, the well-fed fish initiated dispersal faster than the food-deprived fish. The results may indicate state-dependent refuge use and dispersal in C. oxyrinchus.

[Study of the monogenean family Tetraonchidae Bychowsky, 1937 by the scanning electron microscopy].

Hitherto data on the morphology of Tetraonchidae and their eggs obtained with scanning electron microscopy (SEM) were absent in the literature. In the present paper results of SEM study of two genera of Tetraonchidae, Tetraonchus and Salmonchus, are given. Fine morphological traits of eggs, larvae, and definitive individuals of Tetraonchidae, as well as patterns of their attachment to host gill, were established for the first time. A deep penetration of the worms into the branchial epithelium of the host was shown.

Protection against Flavobacterium psychrophilum infection (cold water disease) in Ayu fish (Plecoglossus altivelis) by oral administration of humus extract.

Humic substances are formed during the decomposition of organic matter in humus, and are found in many natural environments in which organic materials and microorganisms have been present. In the present study, oral administration of humus extract to ayu fish (Plecoglossus altivelis) induced effective protection against experimental Flavobacterium psychrophilum infection (cold water disease). Mortality of fish and development of skin lesions, such as erosion and hemorrhages on the skin, gill cover or mouth, were significantly suppressed in fish treated with 10%, 5% or 1% humus extract adsorbed on dry pellets. Although F. psychrophilum was not re-isolated from gills and erosion lesions of the skin of dead fish, bacterial gyrB DNA could be amplified in these specimens from dead fish and surviving control fish using the polymerase chain reaction. The protective effect of the extract was not the results of direct killing of bacteria or antibiotic activity of the extract since no obvious reduction in the bacterial number was observed at 5 times to 5,000 times dilution of the humus extract having pH 5.45 to 7.40. These results clearly show that treating fish with humus extract is effective in preventing cold water disease.

Murres, capelin and ocean climate: inter-annual associations across a decadal shift.

To ensure energy demands for reproduction are met, it is essential that marine birds breed during periods of peak food availability. We examined associations of the breeding chronology of common murres (Uria aalge) with the timing of the inshore arrival of their primary prey, capelin (Mallotus villosus) from 1980 to 2006 across a period of pervasive change in the Northwest Atlantic ecosystem. We also assessed the influence of ocean temperature and the North Atlantic Oscillation (NAO; an index of winter climate and oceanography) on these interactions. We found a lagged linear relationship between variations in murre breeding chronology and the timing of capelin arrival in the previous year. On a decadal level, we found a non-linear threshold relationship between ocean temperature and the timing of capelin arrival and murre breeding. Centennially anomalous cold water temperatures in 1991 generated a marked shift in the timing of capelin spawning inshore and murre breeding, delaying both by more than 2 weeks. By the mid-1990s, ocean temperatures returned to pre-perturbation levels, whereas the temporal breeding responses of capelin and murres were delayed for a decade or more. Oceanographic conditions (temperature, NAO) were poor predictors of the timing of capelin arrival inshore in the current year compared to the previous one. Our findings suggest that knowledge of the timing of capelin availability in the previous year provides a robust cue for the long-lived murres, allowing them to achieve temporal overlap between breeding and peak capelin availability.

Contrasting associations between breeding coloration and parasitism of male Arctic charr relate to parasite species and life cycle stage.

Conspicuous carotenoid ornamentation is considered a signal of individual "quality" and one of the most intensely studied traits found to co-vary with parasitism. Since it has been suggested that only "high quality" individuals have enough resources to express excessive sexual ornaments and resist parasites, current theory struggles to explain cases where the brightest individuals carry the most parasites. Surprisingly little emphasis has been put on the contrasting routes to fitness utilized by different parasite species inhabiting the same host. Using Arctic charr (Salvelinus alpinus) as model species, we hypothesized that skin redness and allocation of carotenoids between skin and muscle (redness ratio) will be positively and negatively associated with parasites using the fish as an intermediate and final host, respectively. Both pigment parameters were indeed positively associated with abundances of parasites awaiting trophic transmission (Diplostomum sp. and Diphyllobothrium spp.) and negatively associated with the abundance of adult Eubothrium salvelini tapeworms. These empirical data demonstrate that contrasting associations between carotenoid coloration and parasite intensities relates to the specific premises of different parasite species and life cycle stages.

Comparative gill and liver pathology of Tilapia zilli, Clarias gariepinus and Neochanna diversus in owan river (Nigeria): Relative ecological risks of species in a pesticide-impacted river.

The dissimilar ecological behavior of species including, extent of mobility and feeding strategies, may predispose them to greater toxic effects. This hypothesis was tested by histological-based assessment of gills and liver of pelagic (Tilapia zilli) and benthic (Clarias gariepinus and Neochanna diversus) fish species of River Owan. The fish species were sampled monthly across seasons from river sites where they were observed to be most abundant. The pathologies were examined from light micrographs, and severity was evaluated by semi-quantitative analyses. Gill pathology of Tilapiazilli showed a moderate occurrence of shortened secondary lamellae, compared with Clarias gariepinus and Neochanna diversus which showed a high incidence of very distinct structural disruptions including epithelial lifting, collapsed secondary lamella structure due to pilaster disruption. Although hepatocellular damage, fibrotic biliary disruptions and parasite incidence were the most evident pathological features in liver of all species, parasite variety and lesion severity differed across species. Principal component analysis (PCA) associated benthic species with more severe gills and liver pathologies, indicating that, benthic species in the Owan riverscape were more at risks compared to pelagic species. As such, we successfully demonstrate that relative ecological risks and potential adverse health effects on fish species could be dependent on ecological behavior and preferences. Conservation efforts particularly for endangered species could be better developed if relative risks and vulnerability of species are understood.

The evolutionary history of sharp- and blunt-snouted lenok (Brachymystax lenok (Pallas, 1773)) and its implications for the paleo-hydrological history of Siberia.

BACKGROUND: Broad-scale phylogeographic studies of freshwater organisms provide not only an invaluable framework for understanding the evolutionary history of species, but also a genetic imprint of the paleo-hydrological dynamics stemming from climatic change. Few such studies have been carried out in Siberia, a vast region over which the extent of Pleistocene glaciation is still disputed. Brachymystax lenok is a salmonid fish distributed throughout Siberia, exhibiting two forms hypothesized to have undergone extensive range expansion, genetic exchange, and multiple speciation. A comprehensive phylogeographic investigation should clarify these hypotheses as well as provide insights on Siberia's paleo-hydrological stability. RESULTS: Molecular-sequence (mtDNA) based phylogenetic and morphological analysis of Brachymystax throughout Siberia support that sharp- and blunt-snouted lenok are independent evolutionary lineages, with the majority of their variation distributed among major river basins. Their evolutionary independence was further supported through the analysis of 11 microsatellite loci in three areas of sympatry, which revealed little to no evidence of introgression. Phylogeographic structure reflects climatic limitations, especially for blunt-snouted lenok above 56 degrees N during one or more glacial maxima. Presumed glacial refugia as well as interbasin exchange were not congruent for the two lineages, perhaps reflecting differing dispersal abilities and response to climatic change. Inferred demographic expansions were dated earlier than the Last Glacial Maximum (LGM). Evidence for repeated trans-basin exchange was especially clear between the Amur and Lena catchments. Divergence of sharp-snouted lenok in the Selenga-Baikal catchment may correspond to the isolation of Lake Baikal in the mid-Pleistocene, while older isolation events are apparent for blunt-snouted lenok in the extreme east and sharp-snouted lenok in the extreme west of their respective distributions. CONCLUSION: Sharp- and blunt-snouted lenok have apparently undergone a long, independent, and demographically dynamic evolutionary history in Siberia, supporting their recognition as two good biological species. Considering the timing and extent of expansions and trans-basin dispersal, it is doubtful that these historical dynamics could have been generated without major rearrangements in the paleo-hydrological network, stemming from the formation and melting of large-scale glacial complexes much older than the LGM.

Net trophic transfer efficiencies of polychlorinated biphenyl congeners to lake whitefish (Coregonus clupeaformis) from their food.

Lake whitefish (Coregonus clupeaformis) were fed rainbow smelt (Osmerus mordax) in four laboratory tanks over a 133-d experiment. At the start of the experiment, 10 to 14 of the fish in each tank were sacrificed, and the concentrations of 40 polychlorinated biphenyl (PCB) congeners within these fish were determined. Polychlorinated biphenyl congener concentrations were also determined in the 15 lake whitefish remaining in each of the four tanks at the end of the experiment as well as in the rainbow smelt fed to the lake whitefish. Each lake whitefish was weighed at the start and the end of the experiment, and the amount of food eaten by the lake whitefish during the experiment was tracked. Using these measurements, net trophic transfer efficiency (gamma) from the rainbow smelt to the lake whitefish in each of the four tanks was calculated for each of the 40 PCB congeners. Results showed that gamma decreased exponentially as log K(OW) for the congeners increased from 6 to 8. Further, gamma averaged 0.70 for the tetrachloro congeners but averaged only 0.45 for the higher chlorinated congeners.

[Mitochondrial DNA genetic differentiation of the muksun Coregonus muksun (Pallas) and related Siberian species of Coregonus (Coredonidae, Salmoniformes)].

Restriction enzyme analysis of the mitochondrial DNA (mtDNA) fragment encoding subunit 1 of the NADH dehydrogenase complex (ND-1) amplified via polymerase chain reaction (PCR) has been used to obtain data on genetic differentiation of muksun Coregonus muksun (Pallas) populations. Population polymorphism with respect to the restriction sites of 18 endonucleases has been described. It has been demonstrated that the muksun is genetically related to the pidschian C. pidschian (Gmelin), its sympatric species in Siberian waters. Analysis of the median network of mtDNA haplotypes has shown that haplotypes of muksun from various Siberian basins form a common group with haplotypes of pidschian of the Arctic Ocean basin, some frequent haplotypes been found in both forms. This raises the question as to the validity of the muksun as a species. Differences within this group of haplotypes are much smaller than those typical of species of the genus Coregonus. The possibility of a hybrid origin of the muksun from a pidschian-like ancestor and species of the cisco-peled (C. sardinella-C. peled) complex is discussed.

[Mitochondrial DNA variation in Pacific capelin (Mallotus villosus catervarius) from the Sea of Okhotsk, inferred from PCR-RFLP analysis].

Four population samples of Pacific capelin Mallotus villosus catervarius (Pennant, 1784) from geographically distant localities in the Sea of Okhotsk, Tauy Bay and the eastern coast of the Sakhalin Island, were examined using PCR-RFLP analysis of three mitochondrial DNA regions (A8/A6/COIII/ND3, ND3/ND4L/ND4, and ND5/ND6). The nucleotide divergence of mtDNA sequences among the samples, as well as the analysis of geographic heterogeneity of the haplotype frequencies and quantitative estimation of genetic differentiation performed by means of AMOVA, showed that the samples examined belonged to one panmictic population. Genealogic analysis of the mtDNA variation structure was carried out. It was demonstrated that the high level of haplotype diversity (0.9639 +/- 0.00015) along with the low level of nucleotide diversity (0.003818 +/- 0.0000003) pointed to the exponential rate of population growth of the capelin from the Sea of Okhotsk, which rather recently in its evolutionary history faced the bottleneck effect.

Density-dependent migratory waves in the marine pelagic ecosystem.

The migration of large aggregations of animals that sweep through the landscape is a phenomenon with large consequences in many ecosystems. It has been suggested that such migrations are mediated by resource depletion. Under this hypothesis it has been shown that simple foraging rules may generate density-dependent migratory waves (DDMW) in which the speed and amplitude increase with animal abundance. We tested these predictions on a 32-year data set of the spatial distribution of the two youngest age groups of a small pelagic schooling fish, the capelin (Mallotus villosus), by the end of their annual feeding migration in the Barents Sea. Our data suggest that the two age groups divided the Barents Sea by forming migratory waves that moved in opposite directions. The aggregation and spatial displacement of these waves increased with increasing age-specific abundance. However, possibly through social interactions, migratory pattern was modified by the abundance of the other age group.

Geographical distribution of Metagonimus yokogawai and M. miyatai in Shizuoka Prefecture, Japan, and their site preferences in the sweetfish, Plecoglossus altivelis, and hamsters.

Sweetfish, Plecoglossus altivelis, obtained from 18 rivers in Shizuoka Prefecture were examined for metacercarial infection of 2 flukes, Metagonimus yokogawai and Metagonimus miyatai. The infection rate and density of metacercariae in the fish were higher in eastern and western regions than in central region of the prefecture. After infection of hamsters with metacercariae derived from the scale, 98.7% of the adult worms obtained from the intestine was found to be M. miyatai. Conversely, from infection with metacercariae from the flesh, 90.0% of the worms was M. yokogawai. Since the worms had no exclusivity in the tissues, we conclude that the flukes have location preference with the former primarily preferring the scale, and the latter the flesh. Fish from two rivers located in adjacent areas in the western region had relatively a higher ratio of M. yokogawai in the scale relative to other rivers, suggesting an intraspecific genetic variation due to geographical isolation. On examination of adult worms in the hamster's intestine, M. yokogawai was mainly located towards the anterior part of the intestine, unlike M. miyatai, suggesting that in mammalian host too, the parasites have site preference.

Nucleotide diversity of Japanese isolates of infectious hematopoietic necrosis virus (IHNV) based on the glycoprotein gene.

Infectious hematopoietic necrosis virus (IHNV), a member of the genus Novirhabdovirus, causes a highly lethal disease of salmonid fish. In the present study, G gene nucleotide sequences of 9 Japanese IHNV isolates obtained from 1971 to 1996 were analyzed to evaluate the genetic diversity and compared with IHNV isolates from North America and Europe. A radial phylogenetic tree revealed 5 major clusters including 3 genogroups (U, M and L) for North American isolates and 1 genogroup for European isolates. Five Japanese isolates from 1971 to 1982 appeared in the cluster for genogroup U, while the remaining Japanese isolates from 1980 to 1996 formed a new genogroup, JRt (Japanese rainbow trout). Maximum nucleotide diversity among the Japanese isolates was 4.5%, which was greater than that within the North American isolates (3.6%), and the degree of nucleotide diversity within Japanese isolates was increased by inclusion of the genogroup JRt isolates. It was concluded that Japanese isolates shared a common source with the genogroup U of the North American isolates and that there were large divergences between Japanese isolates before and after the 1980s.