Salmonidae

The phylogenetic relationships among isolates of the Saprolegnia diclina-Saprolegnia parasitica complex were investigated based on ITS rDNA sequences, and correlated with morphological and physiological characters. The isolates studied belong to five phylogenetically separate clades. The majority of presumed parasitic isolates, mostly isolated from fish lesions, fell within a clade that comprises isolates which has been variously named as S. diclina Type 1, S. parasitica, Saprolegnia salmonis or just as unnamed Saprolegnia sp. Presence of bundles of long-hooked hairs on secondary cysts, high frequency of retracted germination, and oogonia production at 7°C (when occurring) were characteristic of this clade. A single isolate identified as S. diclina Type 2 clustered in a clade along with Saprolegnia ferax isolates. The isolates identified as S. diclina s. str. (S. diclina Type 3) distributed in two clades and appeared closely related to Saprolegnia multispora and to a number of Chilean isolates identified as Saprolegnia australis. The ITS sequences of clade I were almost identical even though the isolates were of diverse geographical origins and showed physiological and morphological differences and variations in their pathogenicity. This suggest these species reproduces clonally even in apparently sexually competent isolates. Adaptation to parasitism in Saprolegnia might have occurred at spore level by the development of long-hooked hairs to facilitate host attachment and selection of a retracting germination. The use of the name S. parasitica should be assigned to isolates of clade I that contained isolates forming cysts with bundles of long-hooked hairs.

Yersinia ruckeri is the causative agent of yersiniosis or enteric redmouth disease leading to significant economic losses in salmonid aquaculture worldwide. Infection may result in a septicaemic condition with haemorrhages on the body surface and in the internal organs. Despite the significance of the disease, very little information is available on the pathogenesis, hampering the development of preventive measures to efficiently combat this bacterial agent. This review discusses the agent and the disease it causes. The possibility of the presence of similar virulence markers and/or pathogenic mechanisms between the Yersinia species which elicit disease in humans and Y. ruckeri is also examined.

We present Wrst insights into the molecular phylogeny of the grayling genus Thymallus (Salmonidae) using sequences from the mitochondrial control region and ATPase6 genes. A suite of analytical approaches were applied for each gene separately and for the combined data. The ATPase6 gene is shown to have a mean divergence rate across the genus of 2.46 times faster than the complete control region. Based on the combined data, four major (internal) clades, presumably originating in the Pliocene, were resolved with high support in all analyses and represented two distinct lineages in the Amur basin, one lineage in all remaining Siberian and Mongolian drainages, and one lineage corresponding to European grayling Thymallus thymallus. The resolution of multiple lineages, from both additional internal and terminal clades, within each major drainage basin underscores the complexity and eVects that Pleistocene hydrological dynamics have had on the distribution of biodiversity in Siberia. 

The extensive phenotypic polymorphism in the European whitefish has triggered evolutionary research in order to disentangle mechanisms underlying diversification. To illuminate the ecological distinctiveness in polymorphic whitefish, and evaluate taxonomic designations, we studied nine Norwegian lakes in three watercourses, which each harboured pairs of divergent whitefish morphs. We compared the morphology and life history of these morphs, documented the extent of genetic differentiation between them, and contrasted the niche use of sympatric morphs along both the habitat and resource axes. In all cases, sympatric morphs differed in the number of gill rakers, a highly heritable trait related to trophic utilization. Individual growth rate, age and size at maturity, diet and habitat use also differed between morphs within lakes, but were remarkably similar across lakes within the same morph. Microsatellite analyses confirmed for all but one pair that sympatric morphs were significantly genetically different, and that similar morphs from different lakes likely have a polyphyletic origin. These results are most compatible with the process of parallel evolution through recurrent postglacial divergence into pelagic and benthic niches in each of these lakes. We propose that sparsely and densely rakered whitefish sympatric pairs may be a likely case of ecological speciation, mediated in oligotrophic lakes with few trophic competitors.

This paper synthesizes tagging studies to highlight the current state of knowledge concerning the behaviour and survival of anadromous salmonids in the marine environment. Scientific literature was reviewed to quantify the number and type of studies that have investigated behaviour and survival of anadromous forms of Pacific salmon (Oncorhynchus spp.), Atlantic salmon (Salmo salar), brown trout (Salmo trutta), steelhead (Oncorhynchus mykiss), and cutthroat trout (Oncorhynchus clarkii). We examined three categories of tags including electronic (e.g. acoustic, radio, archival), passive (e.g. external marks, Carlin, coded wire, passive integrated transponder [PIT]), and biological (e.g. otolith, genetic, scale, parasites). Based on 207 papers, survival rates and behaviour in marine environments were found to be extremely variable spatially and temporally, with some of the most influential factors being temperature, population, physiological state, and fish size. Salmonids at all life stages were consistently found to swim at an average speed of approximately one body length per second, which likely corresponds with the speed at which transport costs are minimal. We found that there is relatively little research conducted on open-ocean migrating salmonids, and some species (e.g. masu [O. masou] and amago [O. rhodurus]) are underrepresented in the literature. The most common forms of tagging used across life stages were various forms of external tags, coded wire tags, and acoustic tags, however, the majority of studies did not measure tagging/handling effects on the fish, tag loss/failure, or tag detection probabilities when estimating survival. Through the interdisciplinary application of existing and novel technologies, future research examining the behaviour and survival of anadromous salmonids could incorporate important drivers such as oceanography, tagging/handling effects, predation, and physiology.

The long-term effects of the sea lice treatment products Excis and Slice on zooplankton communities in a Scottish sea loch were investigated at a commercially operating salmon farm over 31 months. Cypermethrin and emamectin benzoate are the active ingredients in Excis and Slice respectively, which are widely used to control ectoparasitic sea lice on farmed salmon. Excis and Slice treatments did not cause basin-wide effects on the zooplankton community. For both formulations, no adverse affects on zooplankton were detected, instead observed changes in zooplankton abundance and community composition displayed natural seasonal cycles of abundance. Water column concentrations of cypermethrin and emamectin benzoate following sea lice treatments at the fish farm were predicted using models. Cypermethrin concentrations of 3000 ng/l were predicted for short periods immediately after each cage treatment assuming no particle adhesion. The 3-h and 24-h Environmental Quality Standards (EQS) were exceeded for 10 h and 32 h respectively on the second day when five cages were treated. However, cypermethrin concentrations higher than 0.5 ng/l (24-h EQS) were predicted to occur over <2% of the total basin area on each treatment day. The 3-h EQS (16 ng/l) was exceeded in <0.3% of the basin on each treatment day. The concentration of soluble emamectin benzoate present in the water column was predicted from modelled deposition footprints and sediment concentrations to be of order 10 À3 ng/l. Predicted concentrations of both chemicals were generally lower than those causing toxicity to copepods in previous laboratory studies and further support the results of this field study that environmental concentrations of Excis and Slice do not adversely impact zooplankton communities.

Cytokines, produced at the site of entry of a pathogen, drive inflammatory signals that regulate the capacity of resident and newly arrived phagocytes to destroy the invading pathogen. They also regulate antigen presenting cells (APCs), and their migration to lymph nodes to initiate the adaptive immune response. When naive CD4þ T cells recognize a foreign antigen-derived peptide presented in the context of major histocompatibility complex class II on APCs, they undergo massive proliferation and differentiation into at least four different T-helper (Th) cell subsets (Th1, Th2, Th17, and induced T-regulatory (iTreg) cells in mammals. Each cell subset expresses a unique set of signature cytokines. The profile and magnitude of cytokines produced in response to invasion of a foreign organism or to other danger signals by activated CD4þ T cells themselves, and/or other cell types during the course of differentiation, define to a large extent whether subsequent immune responses will have beneficial or detrimental effects to the host. The major players of the cytokine network of adaptive immunity in fish are described in this review with a focus on the salmonid cytokine network. We highlight the molecular, and increasing cellular, evidence for the existence of T-helper cells in fish. Whether these cells will match exactly to the mammalian paradigm remains to be seen, but the early evidence suggests that there will be many similarities to known subsets. Alternative or additional Th populations may also exist in fish, perhaps influenced by the types of pathogen encountered by a particular species and/or fish group. These Th cells are crucial for eliciting disease resistance post-vaccination, and hopefully will help resolve some of the difficulties in producing efficacious vaccines to certain fish diseases.

Using seven pairs of microsatellite markers we studied the relationship between the size of the the candidate groups and the microsatellite paternity testing accuracy by separate breed or the microsatellite paternity appraisal ability by mixed breed. By computer software analysis, the appraisal ability decreased with the candidate groups increase. For the 81 putative parents the microsatellite paternity testing accuracy was 80% and the microsatellite paternity appraisal ability was 78.9%; for the 9 putative parents, the testing accuracy was 93.3% and the appraisal ability was 92.2%. The results indicated that the microsatellite DNA markers can be used for the parentage determination of Hucho taimen Pallas.

The genes of the major histocompatibility complex (MHC) are amongst the most variable in vertebrates and represent some of the best candidates to study processes of adaptive evolution. However, despite the number of studies available, most of the information on the structure and function of these genes come from studies in mammals and birds in which the MHC class I and II genes are tightly linked and class II alpha exhibits low variability in many cases. Teleost fishes are among the most primitive vertebrates with MHC and represent good organisms for the study of MHC evolution because their class I and class II loci are not physically linked, allowing for independent evolution of both classes of genes. We have compared the diversity and molecular mechanisms of evolution of classical MH class II α and class II β loci in farm populations of three salmonid species: Oncorhynchus kisutch, Oncorhynchus mykiss and Salmo salar. We found single classical class II loci and high polymorphism at both class II α and β genes in the three species. Mechanisms of evolution were common for both class II genes, with recombination and point mutation involved in generating diversity and positive selection acting on the peptide-binding residues. These results suggest that the maintenance of variability at the class IIα gene could be a mechanism to increase diversity in the MHC class II in salmonids in order to compensate for the expression of one single classical locus and to respond to a wider array of parasites.

Water quality was measured in 42 streams in the Colebrooke and Upper Bann catchments in Northern Ireland over the period 1990-1998. Despite ongoing pollution control measures, biological water quality, as determined by the invertebrate average score per taxon (ASPT) index, did not improve and there was no appreciable decline in recorded farm pollution incidents. However, the lack of decline in pollution incidents could reflect changes in detection policy, as a greater proportion of incidents were recorded from less polluting discharges such as farm-yard runoff. In contrast, there was an improvement during 1997 and 1998 in annual chemical water quality classification based on exceedence values (90th percentiles) for dissolved oxygen, ammonium and BOD concentrations. In 1998, 11.9% of streams were severely polluted compared to 26.2% in 1990, while the proportion classed as of salmonid water quality, increased from 40.5% in 1990 to 59.6% in 1998. Although water quality in 1996 did not ...

a PCR protocol for the detection of Aeromonas salmonicida in fish by amplification of the fstA (ferric siderophore receptor) gene, Veterinary Microbiology (2007), 3 salmonicida in fish by amplification of the fstA (ferric siderophore 4 receptor) gene 5 6 ABSTRACT 27 28 The aims of the study were to evaluate a new PCR protocol designed to detect 29 Aeromonas salmonicida in fish tissues and to develop a non destructive method for the 30 diagnosis of furunculosis. A set of primers (Fer3, Fer4), flanking a fragment of the fstA 31 gene (coding for the ferric-siderophore receptor) was designed, showing to be sensitive 32 and specific. When compared to PCR methods previously reported, the new protocol 33 recognised all the 69 A. salmonicida strains evaluated, with no cross-reactions with the 34 other bacterial species analysed. Sensitivity assays were performed in fish tissues 35 seeded with serial dilutions of pure cultures of A. salmonicida and mixed cultures of this 36 bacterium with Vibrio anguillarum and A. hydrophila. Detection limits obtained were of 37 60 and 450 bacterial cells 100mg -1 of tissue respectively. Mucus and blood were 38 evaluated in order to develop a non-destructive tool to detect the pathogen. The 39 detection limits in seeded mucus and blood samples were 2.5 x 10 2 and 1 x 10 5 bacterial 40 cells mL -1 , respectively. When the method was used to detect A. salmonicida in 41 asymptomatic wild salmon, 4 samples of mucus and 6 of blood were positive, 42 corresponding to 6 out of the 31 fish examined, whereas only one of the samples 43 resulted positive by culture methods. It is concluded that the PCR protocol evaluated is 44 fast, specific and sensitive to detect A. salmonicida in infected and asymptomatic fish, 45 and will be helpful for the control of the disease through the prompt detection of carriers 46 within fish populations. 47 48 49 50 51 52

The structure and diversity of grayling (Thymallus thymallus) populations have been well studied in most of its native habitat; however the southernmost populations of the Balkan Peninsula remain largely unexplored. The purpose of this study was to assess the genetic diversity of Serbian grayling populations, detect the impact of stocking and provide guidelines for conservation and management. Methods: Eighty grayling individuals were collected from four rivers (Ibar, Lim, Drina and Rzav). The mitochondrial DNA control region (CR; 595 bp of the 3'end and 74 bp of flanking tRNA) and the ATP6 gene (630 bp fragment) were sequenced for 20 individuals (five from each locality). In addition, all individuals were genotyped with 12 microsatellite loci. The diversity and structure of the populations as well as the recent and ancient population declines were studied using specialized software.

Comparative molecular characterization of the regucalcin (RGN) gene in rainbow trout (Oncorhynchus mykiss) and maraena whitefish (Coregonus marena) Abstract The Ca 2? -binding protein regucalcin (RGN) is crucial for the regulation of Ca 2? ion homeostasis and signal transduction of cells. It is involved in the regulation of Ca 2?dependent protein kinases and Ca 2? pump enzymes in cell membranes. Comparative transcriptome analysis in healthy fish of two aquacultured rainbow trout (Oncorhynchus mykiss) lines (BORN, TCO) varying in susceptibility to environmental stress identified significant differences in the expression of the RGN gene. Therefore, we firstly determined the full genomic DNA and cDNA sequence of RGN gene from rainbow trout and comparatively investigated the complete cDNA sequence in another salmonid fish dedicated for local aquaculture, the maraena whitefish (Coregonus marena). The sequence coding region translates for proteins of 298 and 299 amino acids (aa), respectively, indicating a high conservation of RGN proteins (95.7% aa identity) between the two related salmonids. In the second place, we generated RGN gene expression profiles after pathogen (Aeromonas salmonicidae subsp. salmonicida) and temperature (8 and 23°C) challenge in the two rainbow trout lines using salmon microarrays and quantitative RT-PCR. The profiles not only verified initially detected gene expression differences, they also display a tissue specific gene expression in dependence from the stressor and time.

Chile is one of the countries where the development of salmonid farming has been the most successful. The first importation of salmonids in Chile from the northern hemisphere dates back to the late 19th century and the country now ranks as the world second largest producer of farmed salmon. However, the fast increase of infections caused by the bacterium Flavobacterium psychrophilum is a growing concern for this local industry. This pathogen, also recognized as an important problem worldwide, has been first reported in Chile in 1993 and is currently affecting all three cultivated salmonid species: Atlantic salmon (Salmo salar), Coho salmon (Oncorhynchus kisutch) and rainbow trout (O. mykiss).

Mitochondrial and microsatellite DNA markers were applied to infer the phylogeography, intraspecific diversity and dynamics of the distributional history of European grayling ( Thymallus thymallus ) with focus on its central and northern European distribution range. Phylogenetic and nested clade analyses revealed at least four major mtDNA lineages, which evolved in geographical isolation during the Pleistocene. These lineages should be recognized as the basic evolutionary significant units (ESUs) for grayling in central and northern Europe. In addition, and in contrast to previous work on grayling, the results of Bayesian analysis of individual admixture coefficients, two-dimensional scaling analysis and spatial analysis of molecular variance provided evidence for a high level of admixture among major lineages in contact zones between drainages (e.g. the low mountain range of Germany), most likely resulting from glacial perturbations and ancient river connections between drainages during the Pleistocene glaciations. Even within river systems, a high level of differentiation among populations was revealed as indicated by the microsatellite data. Grayling sampled from 29 sites displayed high levels of differentiation (overall F ST = 0.367), a high number of private alleles and high bootstrap support for the genetic distancebased population clusters across 12 loci. We specifically discuss our results in context of phylogeograpic studies on other European freshwater fish species with habitat preferences similar to those of grayling. Our study shows that both large-scale phylogeographical and detailed genetic analyses on a fine scale are mandatory for developing appropriate conservation guidelines of endangered species.

A method for trace analysis of polybrominated diphenyl ethers (PBDE) in fish based on gas chromatography/electron ionization high resolution mass spectrometry (GC/EI-HRMS) was developed, and levels of PBDE were determined in whitefish (Coregonus sp.) from eight Swiss lakes and in rainbow trout (Oncorhynchus mykiss) from four Swiss fish farms. PBDE concentrations (sum of PBDE congeners BDE-28, BDE-47, BDE-99, BDE-100, in filet from whitefish between 36 and 165 ng/g lipid weight (lw) were found, corresponding to wet weight (ww) concentrations of 1.6-7.4 ng/g ww. PBDE contents in filet from farmed rainbow trout were significantly lower than in wild whitefish (12-24 ng/g lw, 0.74-1.3 ng/g ww), and the PBDE congener patterns were different for both species (a higher BDE-47 to BDE-99 ratio for farmed rainbow trout compared to wild whitefish was found). Whitefish PBDE levels [ng/g lw] correlate better with the surface/volume ratio of the respective lakes (r 2 ¼ 0:70) than with other lake properties such as catchment area (size or number of inhabitants) or residence time, suggesting atmospheric deposition as an input pathway for PBDE. Based on an average daily consumption of 20 g whitefish (Switzerland) with a PBDE content of 7.4 ng/g ww (highest PBDE concentration detected in this study), a maximum daily intake of 0.15 lg PBDE was estimated (0.026 lg/day for farmed trout). This number corresponds to the lower end of the estimate for the total PBDE intake of the Nordic consumer of 0.2-0.7 lg/day.

Two previously described lacustrine cisco Coregonus spp. morphs [i.e. a small (<300 mm fork length, L F ), low-gillraker (≤44) morph and a large (≥300 mm L F ), high-gillraker (≥45) morph] from Great Slave Lake, NT, Canada, were found to be synonymous with cisco Coregonus artedi. Geometric body shape did not differ between the two size classes nor could they be differentiated by 24 size-corrected linear measurements, indicating that the two groups had similar phenotypes. Strong, positive correlations between all linear characters and geometric centroid size (a composite variable of fish body length, mass and age) suggested that body morphology changed with age as fish grew. Total gillraker number (N GR ) increased with L F according to: N GR = 36·3 + 0·034L F . Differences in gillraker number and phenotype with age and size were explained by shifts in habitat and trophic resource use. Relative abundance within 0-30, 30-60, 60-90 and >90 m depth strata differed between size classes suggesting that morphology changed when fish shifted their habitat as they grew older. Large C. artedi had lower δ 13 C and slightly higher δ 15 N, indicating greater reliance on pelagic prey resources (i.e. more or larger zooplankton, such as Mysis spp.), compared to small C. artedi , which relied slightly more on benthic prey. Gillraker shape and number have always been used as key diagnostic characters in coregonine taxonomy; based on the findings presented here, ontogenetic shifts should be accounted for in resulting classifications.

This study examined how variability in the abundance and biomass structure of benthic invertebrates affected the feeding choice of the whitefish Coregonus lavaretus on a hard bottom habitat of the brackish Baltic Sea. In general, crustaceans such as Idotea balthica and Gammarus spp. were preferred over molluscs. Although being the most numerous taxon in the invertebrate samples, Mytilus trossulus was the lowest ranking in C. lavaretus food preference. The availability of benthic invertebrate prey set the dietary range of fish but the selectivity largely described fish feeding within this range. There was no clear link between fish predation and the dominance structure of benthic invertebrate communities, suggesting that species composition, abundance and biomass of invertebrate species had no impact on the feeding selectivity of the fish. Thus, while fish predation may not affect the dominant species within a benthic community, due to strong selectivity fish may impose strong pressure on some rarer but highly preferred invertebrate prey species.

Strains of Vibrio salmonicida isolated from Atlantic salmon (Salmo salar) and rainbow trout (Salmo gairdnern) suffering from cold-water vibriosis could be divided on the basis of plasmid profiles into four different categories. Of 32 strains, 19% harbored three plasmids of 24, 3.4, and 2.6 megadaltons (MDa), 69% harbored the 24-and 3.4-MDa plasmids but not the 2.6-MDa plasmid, and 9% harbored only the 24-MDa plasmid. The fourth category, which consisted of only one strain, harbored a plasmid of 10 MDa. In spite of different plasmid patterns, the strains of V. salmonicida were very similar with respect to biochemical reactions. The one-third of the V. salmonicida strains which were serotyped were of the same type. The 50% lethal doses, which were determined by intraperitoneal injection, ranged from 4 x 106 to 1 x 108 CFU per fish. * Corresponding author. t This paper is dedicated to the memory of Emmy Egidius, who died on 3 February 1989.

The Muchalat Inlet, British Columbia, is among the most westerly points at which aquaculture is practiced in Canada. In this paper, we summarise data from over 18000 wild fish sampled at 16 sites over an 8 yr period, between 2004 and 2011. The most prevalent wild species was chum salmon Oncorhynchus keta (82.4%), followed by Chinook O. tshawytscha (10%) and coho O. kisutch (4.3%). However, inter-annual and seasonal variation was evident, and smaller numbers of other Pacific salmon and stickleback species were sporadically observed. A high percentage of wild salmon (~95%) had no sea lice parasites present, with less than 1% of the fish hosting a mobile-stage sea louse. Of the data for which sea lice species were recorded, just over 96% of samples were identified as Lepeophtheirus salmonis. Logistic regression models assessed the association between the presence of lice and a range of independent variables. These models indicated a significant degree of spatial variation, much of whic...

SMITH, J . D., and M. W . LANKESTCR. 1979. Development of swim bladder nematodes (Cystidic,oltr spp.) in their intermediate hosts. Can. J. Zool. 57: 1736-1744. It has been confirmed experimentally that Cysridic.oltr firrionis develops to the infective third-stage in amphipods. The third stage was reached more quickly in Gammtr~~u.sfrrscimtus (5 weeks) than in Hyulclltr trztrJc'rr (6 weeks) at 12-14'C and required 7 weeks to develop in Pontoporritr offinis at 5-7'C. Third-stage larvae were 437 pm long immediately after the second moult and grew up to 4.9 mm long. Clvsridic.01~1 c~ri.stir:omrri developed to the third stage in Mysis rrlicm but not in amphipods. Third-stage larvae were about 720 pm long after the second moult and grew up to 5.7 mm long. Third-stage larvae of C . c.ri,ticornc.r up to 10.8 mm long were found in 0.8% of M . rrlicta from a lake where lake trout were infected with the parasite. Third-stage larvae of C . fcrrionis and C . c,~,isti~o~nc,ri grew considerably in the inter-mediate hosts and the gonads became well developed. Similar development has been reported in other cystidicolids but suggestions that larvae moult to the fourth stage in the intermediate host cannot be accepted. S M I T H . J. D.. et M. W. LAYK[:ST~H. 1979. Development ofswim bladder nematodes (Cy.\ridic.oItr spp.) in their intermediate hosts. Can. J. Zool. 57: 1736-1744. Le developpement dc C'ysticlic~oltr ,firriorzis jusqu'au stade infectieux se fait chez des amphipodes. Les larves atteignent le troisieme stade plus rapidement chez Gtrtnrnnr~~sfi.st~ir~tu.s (5 semaines) que chez Hyrrlrlltr rrzt~c.o (6 sem;~ines) II 12-14°C et il leur faut 7 semaines pour atteindl-e le troisitme stade chez Porlroporuitr trjfinis i 5-7°C. Les larves de 11-oisieme stade ont une 1ongueu1-de 437pm immediatement npres la seconde mue et elles atteignent 4.9mm de longueul-. Cystitlic-olrr c-r.i.trirornc,ri peut atteindl-e le troisieme stade chez My.si.5 relic*/rr. mais pas chez les amphipodes. L.es Iarves de t~.oisieme stade ont environ 720 pm de longueur apres la deuxieme mue et elles croissent jusqu'h 5.7 mm de longueur. Des larves de tl-oisieme stade de D. c.ri.sri~-oinc~~.i de 10.8 mm de longueu~. ont i l e trouvtes chez 0.8% des M . rc~lic,rtr preleves dans un lac ou le touladi etait parasite. Les larves de troisieme stade de C.ftrrionis et de C'. c.~~i.sricornrr.i subissent une croi\s:mce importante chez leur h6te intermediaire et les gonades s'y developpent beaucoup. On sait que d'autres cystidicolides subissent un developpement similaire, mais les theories selon lesquelles les larves se transforment en larves de quatritme stade chez I'h6te intermediaire doivent etl-e rqjetees.

Piscirickettsia salmonis is the bacterium that causes Piscirickettsiosis, a systemic disease of salmonid fish responsible for significant economic losses within the aquaculture industry worldwide. The growth of the bacterium for vaccine formulation has been traditionally accomplished by infecting eukaryotic cell lines, a process that involves high production costs and is time-consuming. Recent research has demonstrated that it is possible to culture pure P. salmonis in a blood containing (cellfree) medium. In the present work we demonstrate the growth of P. salmonis in a liquid medium free from blood and serum components, thus establishing a novel and simplified bacteriological medium. Additionally, the new media reported provides improved growth conditions for P. salmonis, where biomass concentrations of approximately 800 mg cell dry weight L 21 were obtained, about eight times higher than those reported for the blood containing medium. A 2-level full factorial design was employed to evaluate the significance of the main medium components on cell growth and an optimal temperature range of 23-27uC was determined for the microorganism to grow in the novel liquid media. Therefore, these results represent a breakthrough regarding P. salmonis research in order to optimize pure P. salmonis growth in liquid blood and serum free medium. Citation: Henríquez M, González E, Marshall SH, Henríquez V, Gó mez FA, et al. (2013) A Novel Liquid Medium for the Efficient Growth of the Salmonid Pathogen Piscirickettsia salmonis and Optimization of Culture Conditions. PLoS ONE 8(9): e71830.

A substantial amount of research has been done on fish viruses affecting species in aquaculture. This review will focus on the salmonid industry, as this is the most industrialised part of fish farming where vaccines are extensively used. In spite of the amount of research performed, both in commercial companies and in academic organisations, few viral vaccines are licensed. As of today, all fish virus vaccines for sale are based upon inactivated virus or recombinant proteins. No live attenuated or DNA vaccines are currently licensed, but one DNA vaccine against IHN is being tested in controlled field trials in Canada. Vaccines against infectious pancreatic necrosis (IPN) have been sold for many years in Norway and are now also available in Chile. Most of the research on these vaccines has been performed by pharmaceutical companies, and not much information is available as scientific publications. It has also been difficult to establish reproducible IPN challenge models suitable for...

The decision to migrate or not is regarded as genetically controlled for many invertebrate and vertebrate taxa. Here, we show that the environment influences this decision. By reciprocally transplanting brown trout (Salmo trutta L.) between two sections in a river, we show that both migratory and non-migratory behaviour can be environmentally induced; migratory behaviour developed in a river section with high brown trout densities and low specific growth rates, whereas non-migratory behaviour developed in a section with low brown trout densities and high specific growth rates. In a laboratory experiment, we tested the effect of food availability on the development of migratory and non-migratory body morphologies and found that most brown trout became migrants when food levels were low but fewer did so at high food levels. Thus, the decision to migrate seems to be a plastic response, influenced by growth opportunities.

This work deals with mathematical modeling through branching processes. We consider sexually reproducing animal populations where, in each generation, the number of progenitor couples is determined in a non-predictable environment. By using a class of two-sex branching processes, we describe their demographic dynamics and provide several probabilistic and inferential contributions. They include results about the extinction of the population and the estimation of the offspring distribution and its main moments. We also present an application to salmonid populations.

Atypical furunculosis is a problem in farming of salmonids and various other fish species caused by a heterogeneous group of atypical Aeromonas salmonicida strains. Winter ulcer is a disease of salmonids and cod caused by Moritella viscosa, but a number of fish species are susceptible to the infection. Vaccines are available against atypical furunculosis of salmonids, but their efficacy is dependent on the characteristics of the infective strain. Vaccines for non-salmonid fish are currently not commercially available. Furunculosis vaccines for salmon can induce cross protection against some atypical A. salmonicida infections and only in some fish species. Polyvalent injection vaccines based on inactivated bacterial cells are available against winter ulcer disease of salmonids. Outbreaks of winter ulcer disease in vaccinated salmon are, however, continuously reported.

Flavobacterium psychrophilum causes serious fish diseases such RTFS and BCWD, affecting the aquaculture industry worldwide. Commercial vaccines are not available and control of the disease depends on the use of antibiotics. Reliable methods for detection and identification of different isolates of this bacterium could play an important role in the development of good management strategies. The aim of this study was to identify genetic markers for discrimination between isolates. A selection of eight VNTRs from 53 F. psychrophilum isolates from Norway, Chile, Denmark and Scotland were analyzed. The results were compared with previous work on the same pathogen using MLST for genetic differentiation. The VNTR analysis gave a separation between the F. psychrophilum isolates supporting the results of previous MLST work. A higher diversity was found among the Chilean isolates compared to those from Norway, which suggests a more homogenous reservoir in Norway. Transgenerational transmission of F. psychrophilum from other countries, exporting salmon embryos to Chile, may explain the differences in diversity. The same transmission mechanisms could also explain the wide geographical distribution of identical isolates in Norway. But, this could also be a result of movement of smolts and embryos. The selected VNTRs are stable genetic markers and no variation was observed after several passages on agar plates at different temperatures. These VNTRs are important additions for genotyping of F. psychrophilum isolates. Future studies on VNTRs of F. psychrophilum should include isolates from more host species from a wider geographical area. To get a more robust genotyping the VNTRs should be used in concert with MLST. Future studies of isolates with high and low virulence should focus on identifying virulence markers using VTNRs and MLST.

Establishing exposure to contaminants within a given environment is often difficult for fish species with large home ranges. Chemical analyses of muscle or visceral tissue are useful indicators of recent exposure, but depuration, metabolic transformation, and tissue redistributions can alter temporal resolution. Otoliths are metabolically stable and thus provide complete chemical records within their calcified tissues that, when coupled to the annular structure, can provide temporal resolution for exposure to trace metals. Otoliths from bull trout, cutthroat trout, and mountain whitefish from an area rich in seleniferous soils and with active coal mining activity were analyzed for selenium to determine any history of exposure to elevated levels of selenium. Selenium concentrations in otolith primordia tended to be low, indicating that these fish emerged in low selenium areas. Later life stages showed peaks of high Se concentrations, suggesting that individuals moved into areas of increased selenium later in life. Individuals captured from the same area had a wide variety of selenium exposure profiles, indicating that these fish do not move en masse into and out of high-selenium areas. Year-to-year variability of selenium exposure patterns within an otolith suggests inconsistent utilization of high-and low-selenium areas by the individual. The inconsistent exposure profiles for these fish, in addition to their home range of tens of kilometres, indicate that soft tissue concentrations, while useful indicators of recent exposure, cannot be relied upon to provide a life history recording of exposure.

A 20-day experiment was conducted to compare the accumulation, distribution, and toxicological effects of dietary As, as arsenate, in lake whitefish (LWF, Coregonus clupeaformis) and lake trout (LT, Salvelinus namaycush). Results of this experiment were used to design an experiment of longer duration in which one of the fish species was selected and exposed three times per week to lower dietary As doses. In the present study each treatment group was exposed to a combination of one of three doses of As (0, 100, or 1000 mg Asyg) and one of two types of diet, no brine shrimp (NS) or with brine shrimp (WS) for a total of eight dosing events. Brine shrimp were added to determine whether their presence enhanced consumption of As-contaminated food. Modified feeding behavior occurred in both fish species fed As contaminated diets, with the exception of the 100 mg Asyg NS food. Brine shrimp addition did not affect feed consumption of the As contaminated diets. Significant As accumulation occurred in stomach, pyloric caeca, intestine, liver, kidney, and gallbladder, but not in bile or muscle. As exposure did not have a significant effect on hepatic and renal metallothionein concentrations. Concentrations of lipid peroxides were only significantly elevated in the plasma of LT fed the 1000 mg Asyg WS food. Liver somatic indices decreased significantly in both species, whereas hematological parameters were not affected in either species. Histological lesions occurred in gallbladder, liver, kidney, pyloric caeca and intestine from LWF. These lesions were not observed in LT; however, gallbladders were not examined in this species. Weight gain was lower in both species fed As contaminated diets, however, condition factors were not affected.ERROR 1063: Character not permitted in a command.ep ᮊ

The directly transmitted viviparous gyrodactylids have high species richness but low morphological and biological diversity, and many species are recorded from only a single host. They therefore constitute a guild of species ideal for studies of the evolutionary significance of host specificity. The group has the widest host range of any monogenean family, being found on 19 orders of bony fish. However, individual species range from narrowly specific (71% of 402 described species recorded from a single host) to extremely catholic (Gyrodactylus alviga recorded from 16 hosts). Gyrodactylid-host interactions extend from 60 mya (G. lotae, G. lucii) down to 150 years (G. derjavini on Oncorhynchus mykiss). Co-evolution with the host is comparatively rare within the gyrodactylids, but host switching or ecological transfer is common, and has been facilitated by the mixing of fish strains that followed glaciation. In this review, we consider the factors responsible for gyrodactylid specificity patterns, using examples from our work on salmonid gyrodactylids including G. salaris, responsible for major epidemics on wild Atlantic salmon (Salmo salar) in Norway since 1975, and G. thymalli from grayling and G. derjavini from trout.

Fisheries catches worldwide have shown no increase over the last two decades, while aquaculture has been booming. To cover the demand for fish in the growing human population, continued high growth rates in aquaculture are needed. A potential constraint to such growth is infectious diseases, as disease transmission rates are expected to increase with increasing densities of farmed fish. Using an extensive dataset from all farms growing salmonids along the Norwegian coast, we document that densities of farmed salmonids surrounding individual farms have a strong effect on farm levels of parasitic sea lice and efforts to control sea lice infections. Furthermore, increased intervention efforts have been unsuccessful in controlling elevated infection levels in high salmonid density areas in 2009 -2010. Our results emphasize host density effects of farmed salmonids on the population dynamics of sea lice and suggest that parasitic sea lice represent a potent negative feedback mechanism that may limit sustainable spatial densities of farmed salmonids.

Colossoma macropomum, locally called tambaqui, is a freshwater migratory teleost that shows good tolerance to oxygen and pH changes in water, and both chemical-physical parameters change markedly during the day time and seasonal water level oscillations in the Amazon Basin. In order to obtain a general view about the basal levels of antioxidants in different tissues of wild tambaqui, enzymatic (superoxide dismutase and catalase) and non-enzymatic (alpha-tocopherol, beta-carotene, and glutathione) antioxidants and lipid peroxidation levels were assessed in the liver, blood and plasma of ten specimens collected during the dry season (September) in a pond near Manaus-AM, Brazil. Superoxide dismutase, catalase, and lipid peroxidation levels were high in the liver and low in the blood and plasma. Confirming previous results on tambaqui, catalase was detected in the blood of one specimen only. beta-Carotene was not found in any analyzed tissue, while alpha-tocopherol was found only in the...

Chitinolytic activity, nutrient uptake and intestinal barrier functions were investigated in gut tissues of Arctic charr (Salvelinus alpinus) fed iso-nitrogenous diets based on fish meal, with or without inclusion of zygomycetes (Rhizopus oryzae). We found that gut tissue of Arctic charr had significant chitinase activity, of both endo-and exo-chitinase iso-forms. Moreover, the distribution pattern along the GI tract of Arctic charr differed between endo-chitinase and exo-chitinase. The endo-chitinase activity in stomach tissue and in the distal intestine was several hundred-fold higher than the exo-chitinase activity in stomach tissue. The greatest exo-chitinase activity was found in the distal intestine. The zygomycete-based diet resulted in higher chitinolytic activity in gut tissue compared to the fish meal-based diet. Disturbed intestinal integrity and increased uptake rate of the amino acid lysine were observed in the distal, but not proximal, intestine of fish fed the zygomycete-based feed.

The objective of this study was to describe the morphometry and motility parameters of vendace (Coregonus albula) spermatozoa. Morphometric parameters of vendace sperm head and tail were of values similar to rainbow trout. The effects of pH, sodium, potassium and calcium ion concentrations on computer-assisted sperm analysis (CASA) sperm motility characteristics were tested. Vendace sperm was motile in a wide pH range of 6.0-10.5 with the optimum pH established at 9.0. Increases in potassium and calcium ions caused decreases in the percentage of motile sperm. The CASA parameters and erratic sperm movement pattern of vendace spermatozoa were similar to whitefish (C. lavaretus) sperm motility, suggesting that there is a coregonid-specific sperm motility pattern.

The Chilean salmonid industry was developed by importing breeding materials, a practice still in effect due to deficits in the national supply of roe. Importation of breeding materials is often associated with the transmission of pathogens. The objectives of this study were to compare the infectious pancreatic necrosis virus (IPNV) isolates from Chile to those of European origin and to determine the diversity of the Chilean IPNV. The VP2 genes of IPNV from Chilean fish (whose eggs originated from Scotland, Iceland and Norway) were compared to isolates from fish in Norway and Ireland. The results show that the isolates are identical (97–99 %) and cluster into one genogroup. Our findings support previous reports of association between the trade-in breeding materials and transmission of pathogens. Furthermore, our results demonstrate the genotypic diversity of Chilean IPNV isolates. These findings have important implications for IPNV disease diagnosis and control in Chile.

We describe an emerging initiative - the &#39;Functional Annotation of All Salmonid Genomes&#39; (FAASG), which will leverage the extensive trait diversity that has evolved since a whole genome duplication event in the salmonid ancestor, to develop an integrative understanding of the functional genomic basis of phenotypic variation. The outcomes of FAASG will have diverse applications, ranging from improved understanding of genome evolution, to improving the efficiency and sustainability of aquaculture production, supporting the future of fundamental and applied research in an iconic fish lineage of major societal importance.

Whitefish, genus Coregonus, show exceptional levels of phenotypic diversity with sympatric morphs occurring in numerous postglacial lakes in the northern hemisphere. Here, we studied the effects of human-induced eutrophication on sympatric whitefish morphs in the Swiss lake, Lake Thun. In particular, we addressed the questions whether eutrophication (i) induced hybridization between two ecologically divergent summer-spawning morphs through a loss of environmental heterogeneity, and (ii) induced rapid adaptive morphological changes through changes in the food web structure. Genetic analysis based on 11 microsatellite loci of 282 spawners revealed that the pelagic and the benthic morph represent highly distinct gene pools occurring at different relative proportions on all seven known spawning sites. Gill raker counts, a highly heritable trait, showed nearly discrete distributions for the two morphs. Multilocus genotypes characteristic of the pelagic morph had more gill rakers than genotypes characteristic of benthic morph. Using Bayesian methods, we found indications of recent but limited introgressive hybridization. Comparisons with historical gill raker data yielded median evolutionary rates of 0.24 haldanes and median selection intensities of 0.27 for this trait in both morphs for 1948–2004 suggesting rapid evolution through directional selection at this trait. However, phenotypic plasticity as an alternative explanation for this phenotypic change cannot be discarded. We hypothesize that both the temporal shifts in mean gill raker counts and the recent hybridization reflect responses to changes in the trophic state of the lake induced by pollution in the 1960s, which created novel selection pressures with respect to feeding niches and spawning site preferences.