Whole-Genome Resequencing Reveals Polygenic Signatures of Directional and Balancing Selection on Alternative Migratory Life Histories.

Migration in animals and associated adaptations to contrasting environments are underpinned by complex genetic architecture. Here, we explore the genomic basis of facultative anadromy in brown trout (Salmo trutta), wherein some individuals migrate to sea while others remain resident in natal rivers, to better understand how alternative migratory tactics (AMTs) are maintained evolutionarily. To identify genomic variants associated with AMTs, we sequenced whole genomes for 194 individual trout from five anadromous-resident population pairs, situated above and below waterfalls, in five different Irish rivers. These waterfalls act as natural barriers to upstream migration and hence we predicted that loci underpinning AMTs should be under similar divergent selection across these replicate pairs. A sliding windows based analysis revealed a highly polygenic adaptive divergence between anadromous and resident populations, encompassing 329 differentiated genomic regions. These regions were associated with 292 genes involved in various processes crucial for AMTs, including energy homeostasis, reproduction, osmoregulation, immunity, circadian rhythm and neural function. Furthermore, examining patterns of diversity we were able to link specific genes and biological processes to putative AMT trait classes: migratory-propensity, migratory-lifestyle and residency. Importantly, AMT outlier regions possessed higher genetic diversity than the background genome, particularly in the anadromous group, suggesting balancing selection may play a role in maintaining genetic variation. Overall, the results from this study provide important insights into the genetic architecture of migration and the evolutionary mechanisms shaping genomic diversity within and across populations.

Molecular cloning and characterization of Cu-Zn superoxide dismutase (sod1) gene in brown trout and its expression in response to acute aquaculture stressors.

Aquaculture species are often exposed to acute stressors such as low water levels and handling during routine aquaculture procedures. This might result in oxidative stress by the increased reactive oxygen species (ROS)' production (e.g., superoxide anion). The harmful effects of ROS are eliminated by a defense system, referred antioxidant defense system (ADS). sod1 is the first gene involved in the ADS. Therefore, we cloned and characterized the open reading frame of the sod1 in brown trout. Then, we determined the effects of low water level and handling stress on sod1 mRNA expression in the liver and gills at 0 min, 1 and 2 h. The total RNA isolated was used to synthesize cDNA for RT-qPCR analysis. Phylogenetic tree, identity/similarity percentages, genomic organization, and conserved gene synteny analyses were applied to characterize Sod1/sod1. While low water level stress upregulated sod1 expression in the liver compared to the control group, no significant differences were observed in the gills between experimental groups. However, brown trout differently responded to handling stress at different time intervals in both tissues. Transcriptional differences were also noted between the sexes. This study contributes to the current understanding of the molecular mechanism between oxidative stress and ADS.

High summer macrophyte cover increases abundance, growth, and feeding of juvenile Atlantic salmon.

Aquatic habitats are severely threatened by human activities. For anadromous species, managing freshwater habitats to maximize production of more, larger juveniles could improve resilience to threats in marine habitats and enhance population viability. In some juvenile salmonid habitats, complexity created by large substrates provides resources and reduces competitive interactions, thereby promoting juvenile production. In lowland rivers, which lack large substrates, aquatic plants might provide similar complexity and enhance fish productivity. To test the influence of aquatic plants on juvenile Atlantic salmon and sympatric brown trout in a lowland river, we directly manipulated the cover of the dominant macrophyte, Ranunculus, in nine sites during summer and autumn for two years. We quantified the abundance, site retention and growth of salmon and trout under high, medium or low Ranunculus cover. To investigate the effects of Ranunculus cover on feeding opportunities and interspecific competition, we quantified available prey biomass and body size, fish diet composition and compared dietary niche overlap. Experimentally increased Ranunculus cover supported higher salmon abundance in summer and autumn, and higher site retention and growth of salmon in summer. Trout abundance and growth were not influenced by Ranunculus cover, but trout site retention doubled in high, relative to low, cover sites. Despite the weak effects of Ranunculus cover on prey availability, salmon and trout inhabiting high cover sites consumed larger prey and a higher biomass of prey. Furthermore, dietary niche overlap was lower in high, relative to low, cover sites, suggesting that abundant Ranunculus reduced interspecific competition. This field experiment shows that high Ranunculus cover can support more and better growing juvenile salmon, and facilitate foraging and co-existence of sympatric salmonid species. Maintaining or enhancing natural macrophyte cover can be achieved through sympathetic in-river and riparian vegetation management and mitigating pressures on them, such as sediment inputs and low flows, or through planting. Further research should test whether macrophyte cover benefits propagate to subsequent life stages, particularly juvenile overwintering associated with high mortality. This knowledge, in combination with our findings, would further clarify whether beneficial juvenile habitat can improve the viability of at-risk salmonid populations. Overall, our findings suggest that the aims of river restoration might be achieved through promotion of in-stream aquatic vegetation.

Temperature regime during embryogenesis alters subsequent behavioural phenotypes of juvenile brown trout.

Climate warming imposes a serious threat, especially to freshwater ecosystems in temperate and (sub)polar regions, which are often dominated by cold-adapted ectotherms. Although relatively intense warming during winter is common across the climatic regions, comparably little focus has been put on the organismal impacts of winter warming. Embryonic development, which is exceptionally susceptible to ambient temperature, occurs during winter in various freshwater ectotherms. Yet, our knowledge of the effects of increased temperature during embryogenesis on later life stages is limited. Using brown trout (Salmo trutta), we examined how a 1.5°C temperature increase from fertilization to hatching affects various traits at the onset of the free-swimming stage (i.e. a comparison between 3.5 and 5.0°C treatments). Although all hatchlings were kept at the same temperature (7.0°C) from hatching to the onset of the free-swimming stage for about two months, the temperature increase during embryogenesis substantially reduced key ecological behaviours, i.e. activity and exploration levels, at the onset of the free-swimming stage despite only marginal temperature effects on morphological and physiological traits at this stage. Given the importance of behavioural traits in early growth and survival, our study suggests a likely pathway through which subtle changes in mean winter temperature affect early fitness.

Distribution and prevalence of the myxozoan parasite Tetracapsuloides bryosalmonae in northernmost Europe: analysis of three salmonid species.

Global climate change is altering the abundance and spread of many aquatic parasites and pathogens. Proliferative kidney disease (PKD) of salmonids caused by the myxozoan Tetracapsuloides bryosalmonae is one such emerging disorder, and its impact is expected to increase with rising water temperature. Yet, the distribution and prevalence of T. bryosalmonae in Northern Europe remain poorly characterized. Here, we studied 43 locations in 27 rivers in northernmost Norway and Finland to describe T. bryosalmonae infection frequency and patterns in 1389 juvenile salmonids. T. bryosalmonae was discovered in 12 out of 27 rivers (44%) and prevalence ranged from 4.2 to 55.5% in Atlantic salmon and from 5.8 to 75% in brown trout among infected rivers. In sympatric populations, brown trout was more frequently infected with T. bryosalmonae than was salmon. Age-specific parasite prevalence patterns revealed that in contrast to lower latitudes, the infection of juvenile fish predominantly occurs during the second summer or later. Temperature monitoring over 2 yr indicated that the mean water temperature in June was 2.1 to 3.2°C higher in rivers containing T. bryosalmonae compared to parasite-free rivers, confirming the important role of temperature in parasite occurrence. Temporal comparison in T. bryosalmonae prevalence over a 10 yr period in 11 rivers did not reveal any signs of contemporary parasite spread to previously uninfected rivers. However, the wide distribution of T. bryosalmonae in rivers flowing to the Barents Sea indicates that climate change and heat waves may cause new disease outbreaks in northern regions.

Surfing the tide: Homeward migration of sea trout (Salmo trutta) in a Patagonian river.

This study evaluates the influence of marine and freshwater conditions on the timing of river entry and upstream migration of sea trout (Salmo trutta) in the Grande River of Tierra del Fuego, Patagonia. We analysed the in-river catch-and-release records from a group of fishing lodges that dominate the Grande River fishery during January-April 2008 (n = 5029 fish) as a function of environmental variables: tidal amplitude, stage in the lunar cycle, river discharge, and river water temperature along the homeward migration season. We discuss the value of the daily catch rate as an abundance index in the Grande river, then analyse the temporal structure of the tidal cycle in the Grande River estuary, a macro-tidal environment with a mean tidal amplitude of 5.7 m, and analyse the fit of a generalized additive model to trout catches on a daily basis in four sections along the river to identify the environmental variables that may affect trout abundance throughout the homeward migration. Fish catches in each section of the river were differentially affected by specific environmental variables: tidal amplitude had a positive and significant effect on catches in the lower river sections, whereas water temperature and river discharge significantly affected catches in upper sections (positive effect of temperature; negative effect of discharge). Catches in the lower section clearly reflect the river entry stage of the homeward migration, with a bi-modal shape significantly correlated with the tidal cycle. The first peak was composed mainly of larger multi-sea-winter trout that move upstream, whereas the second one had a wider range of fish lengths, including a large proportion of small and maybe nonreproductive trout that overwinter in the lower river. Based on our results, we conclude that the large tides in the Grande River estuary strongly affect the river entry timing of sea trout. The underlying mechanisms of this effect may be a combination of increased olfactory recognition and increased tidal transport modulated by the seasonal tidal cycle, which operates on trout during coastal migration to produce the pulses observed in the Grande River sea trout run. In the middle and upper sections of the river, where the tidal effect at river entry was dissipated as upstream migration progressed, trout catches increased with water temperature and decreased with river discharge, which may operate through their influence on in-river migration rate and abundance, but also through changes in catchability.

Habitat associations of rainbow trout Oncorhynchus mykiss and brown trout Salmo trutta fry.

Habitat restoration activities continue to increase in large rivers, but many of these projects focus on improving juvenile or adult habitats. Incorporating the habitat associations of fry into restoration designs will allow for broader successes from restoration for all life stages and may be useful for either multispecies or specific-species management. This study investigated the habitat associations of rainbow trout Oncorhynchus mykiss and brown trout Salmo trutta fry in the upper Colorado River, focusing on the mean substrate size (D50 ), velocity (m s-1 ), depth (m) and presence of wood in near-shore habitats. S. trutta and O. mykiss were found in higher numbers in fry sites with a D50 of 151 mm (ranging from 96 to 206 mm), velocities ranging from 0.20 to 0.23 m s-1 and depths ranging from 0.17 to 0.18 m. Although there was considerable overlap in habitat associations between the two species, there may be opportunities for single-species management, if this is a goal of such restoration activities, by adjusting design criteria based on differing habitat associations. In addition, the results suggest that including larger particle sizes in near-shore habitats and upstream of fry sites could decrease Tubifex tubifex habitat and thereby fry infection severity by reducing exposure to Myxobolus cerebralis. Stocking, interspecific competition and/or the presence of pathogens can affect fry habitat associations and cause deviations from demonstrated suitability indices. As such, evaluating system-specific differences in habitat associations may allow future habitat restoration activities to be more effective.

Influence of laurel (Laurus nobilis) essential oil on gut function of Black Sea salmon (Salmo labrax) juveniles.

The present work investigated the effects of dietary incorporation of laurel (Laurus nobilis) essential oil on the zootechnical performance and digestive physiology of juvenile Black Sea salmon (Salmo labrax). In this trial, 15 fiberglass tanks (39 × 39 cm square and 33 cm high) were used. Forty-five fish (3.52 ± 0.01 g) were placed randomly per tank. Fish were fed for 90 days with the diet containing 50, 100, 200, or 400 mg kg-1 laurel (Laurus nobilis) essential oils, respectively. The work was performed in the recirculating aquaculture system (RAS) operating with freshwater. Fish were manually fed 3% level of live weight during the experiment period. Final weight (FW), weight gain (WG), feed conversion rate (FCR), and specific growth rate (SGR) were not affected by laurel essential oil supplementation. Dietary laurel essential oil (50 mg kg-1) affected positively the surface area of fish intestinal villus that required for digestion. Both villus height and villus width were affected positively in fish fed with 50 mg laurel essential oil kg-1. While incorporation with 100 mg laurel essential oil kg-1 increased the total α-amylase enzyme, 50 mg laurel kg-1 increased lipase enzyme. Moreover, 50 mg laurel essential oil kg-1 increased lactic acid bacteria (LAB) count in fish. Besides, 50 mg laurel essential oil kg-1 reduced the number of total coliform and E. coli.

Associations between metabolic traits and growth rate in brown trout (Salmo trutta) depend on thermal regime.

Metabolism defines the energetic cost of life, yet we still know relatively little about why intraspecific variation in metabolic rate arises and persists. Spatio-temporal variation in selection potentially maintains differences, but relationships between metabolic traits (standard metabolic rate (SMR), maximum metabolic rate (MMR), and aerobic scope) and fitness across contexts are unresolved. We show that associations between SMR, MMR, and growth rate (a key fitness-related trait) vary depending on the thermal regime (a potential selective agent) in offspring of wild-sampled brown trout from two populations reared for approximately 15 months in either a cool or warm (+1.8°C) regime. SMR was positively related to growth in the cool, but negatively related in the warm regime. The opposite patterns were found for MMR and growth associations (positive in warm, negative in the cool regime). Mean SMR, but not MMR, was lower in warm regimes within both populations (i.e. basal metabolic costs were reduced at higher temperatures), consistent with an adaptive acclimation response that optimizes growth. Metabolic phenotypes thus exhibited a thermally sensitive metabolic 'floor' and a less flexible metabolic 'ceiling'. Our findings suggest a role for growth-related fluctuating selection in shaping patterns of metabolic variation that is likely important in adapting to climate change.

Social status modulates the behavioral and physiological consequences of a chemical pollutant in animal groups.

The social environment (i.e., the suite of social interactions that occur among individuals that can result in variation in social ranks) is a commonly overlooked aspect of biology when scientists evaluate the effects of chemical contaminants. The social environment, however, represents the arena in which individual-level performance shapes group- or population-level outcomes and may therefore mediate many of the ultimate consequences of chemicals for wildlife. Here, we evaluated the role that the social environment plays in determining the consequences of pollutant exposure. We exposed groups of juvenile brown trout (Salmo trutta) to an emerging pharmaceutical pollutant that is commonly detected in freshwaters (the benzodiazepine, oxazepam) and allowed them to form dominance hierarchies. Exposure affected dominant and subordinate fish differently, causing fish to become less aggressive at high doses and subordinate fish to become more competitively successful at low doses. These perturbations had further consequences for growth, fin damage, and survival. Exposure also modulated physiological stress in the hierarchy, and social status itself affected how much oxazepam was absorbed in tissues, potentially creating a dynamic feedback loop that further influences the asymmetric effects of exposure on differing social statuses. Many effects followed a "U-shaped" dose-response curve, highlighting the importance of nonlinear, low-dose effects. Altogether, we show that social structure in animal groups can interact with and modulate the effects of an environmental contaminant. We underscore the need to account for an organism's natural ecological context, including their social environment, in future experiments and environmental risk assessments to predict the effects of chemical contaminants on wildlife.

Sea lice infections on wild Atlantic salmon and sea trout in the River Tamar, UK: a temporal study.

Sea lice are amongst the most ecologically and economically damaging parasites of farmed salmonids globally. Spill-over from aquaculture can increase parasite pressure on wild fish populations, but quantifying this effect is challenging due to the relative paucity of data available on 'natural' salmonid louse burdens in the absence of aquaculture, particularly for Atlantic salmon Salmo salar. Here, wild Atlantic salmon and sea trout S. trutta were screened at the tidal limit of the River Tamar (UK) for the presence of sea lice. During 2013 and 2015, the prevalence of sea lice ranged from 41 (n = 361) to 60% (n = 275) and 55 (n = 882) to 58% (n = 800) in Atlantic salmon and sea trout, respectively. All sea lice collected were identified as Lepeophtheirus salmonis. Mean L. salmonis infection intensity across the study period was 5.84 (range: 1-66) in Atlantic salmon and 6.45 (range: 1-37) in sea trout. Infection intensity was positively correlated with the amount of external damage present for both fish species. Given that the fish were examined when returning to freshwater, the lice burdens obtained may represent an underestimate. Nevertheless, these data provide important baseline information on 'natural' sea louse infections in South West England, which has been proposed as a potential region for aquaculture development.

Using PIT-tags and portable antennas for quantification of fish movement and survival in streams under different environmental conditions.

While PIT-tag tracking using mobile antennas is being increasingly used to study fish movement and survival in streams, little is known about the limitations of the method, especially over longer periods of time and under different environmental settings. We used 6 years of data combining tagging, mobile antenna tracking and recaptures of Salmo trutta in multiple small streams in the Lake Lucerne drainage area in Switzerland to evaluate the relative importance of different environmental and intrinsic factors affecting the efficiency of the method. Our study system and experimental design allowed us to accurately verify the continuous presence and survival of recaptured fish in the stream after tracking, which meant that we could estimate detection probability with high confidence. The mean detection probability of tagged trout was 43%, but we found that fish length had a strong negative effect on detection probability, especially in males. Multivariate axes of stream environmental features did not predict efficiency but stream width alone was significantly positively correlated with efficiency. Additionally, stream temperature when tracking had a positive effect on fish detectability. Tag loss at recapture was globally rare (<8%) but common in large postspawn females (>30%). Based on the escape response of fish after detection, we could estimate the proportion of ghost tags, which reached a plateau of around 80% 2 years after tagging. We finally showed that our models of tag loss, fish detection and escape response are needed to interpret detection events. Our results highlight that individual variation in detection probability and tag loss is high and has to be considered for analysis.

Continuous outmigration and sequential encountering of environmental cues are important for successful homing of hatchery-reared, anadromous brown trout Salmo trutta.

When rehabilitating and reintroducing trout Salmo trutta in rivers, it is a goal that as many as possible survive, home and form self-sustaining populations. Hatchery-reared, anadromous S. trutta have significant lower ability to return to the area where they were raised if (a) transported in a closed tank to sea and released 5 km from the River Imsa, relative to those that were (b) transported when swimming in a partly submerged tank with sea water run-through, while being slowly towed by a boat the same distance or (c) released at the outlet of the River Imsa. Thus, if deprived from environmental cues during part of the way, they lose their ability to home.

Differences in growth between offspring of anadromous and freshwater brown trout Salmo trutta.

In this study, individual growth of juvenile offspring of anadromous and freshwater resident brown trout Salmo trutta and crosses between the two from the River Imsa, Norway, was estimated. The juveniles were incubated until hatching at two temperatures (±S.D.), either 4.4 ± 1.5°C or 7.1 ± 0.6°C. Growth rate was estimated for 22 days in August-September when the fish on average were c. 8 g in wet mass, and the estimates were standardized to 1 g fish dry mass. Offspring of anadromous S. trutta grew better at both 15 and 18°C than offspring of freshwater resident S. trutta or offspring of crosses between the two S. trutta types. This difference appears not to result from a maternal effect because anadromous S. trutta grew better than the hybrids with anadromous mothers. Instead, this appears to be an inherited difference between the anadromous and the freshwater resident fish lending support to the hypothesis that anadromous and freshwater resident S. trutta in this river differ in genetic expression. Egg incubation temperature of S. trutta appeared not to influence the later growth as reported earlier from the studies of Atlantic salmon Salmo salar.

Short-term homeostatic regulation of blood/interstitial fluid Ca2+ concentration by the scales of anadromous sea trout Salmo trutta L. during smoltification and migration.

The elasmoid scales of anadromous sea trout Salmo trutta L. represent a significant internal reservoir of Ca2+ . Although more is known about long-term remodelling of scales in response to calciotropic challenges encountered during smoltification and migration, very little is known about the contribution made by scales to the short-term, minute-to-minute regulation of Ca2+ homeostasis in the extracellular fluid (ECF) during these phases of the life cycle. This gap in the knowledge is partly due to the technical challenges involved in measuring small Ca2+ fluxes around the scales of live fish in real time. Here, this study describes exfoliating, mounting and culturing scales and their resident cells from parr, smolt and adult sea trout from a freshwater environment, as well as from adult sea trout caught in sea or brackish water. All the scales were then examined using an extracellular, non-invasive, surface-scanning Ca2+ -sensitive microelectrode. The authors quantified the Ca2+ fluxes, in the absence of any systemic or local regulators, into and out of scales on both the episquamal and hyposquamal sides under different extracellular calcemic challenges set to mimic a variety of ECF-Ca2+ concentrations. Scales from the life-cycle stages as well as from adult fish taken from sea, brackish or fresh water all showed a consistent efflux or influx of Ca2+ under hypo- or hypercalcemic conditions, respectively. What were considered to be isocalcemic conditions resulted in minimal flux of Ca2+ in either direction, or in the case of adult scales, a consistent but small influx. Indeed, adult scales appeared to display the largest flux densities in either direction. These new data extend the current understanding of the role played by fish scales in the short-term, minute-to-minute homeostatic regulation of ECF-Ca2+ concentration, and are similar to those recently reported from zebrafish Danio rerio scales. This suggests that this short-term regulatory response might be a common feature of teleost scales.

Heavy metal and metalloid accumulation in wild brown trout (Salmo trutta L., 1758 complex, Osteichthyes: Salmonidae) from a mountain stream in Sardinia by ICP-OES.

 This paper reports heavy metal and metalloid accumulation in wild brown trout (Salmo trutta L., 1758 complex) raised in freshwater and uncontaminated Sardinia system (Italy). Metals are widespread pollutants of aquatic systems, and their contamination can originate from anthropogenic activities such as industrial waste, agricultural and domestic environments, and geochemical release. Fish has a relevant position within the human diet; moreover, fishes can accumulate metals, making them a valuable tool as biomarkers for risk assessment studies. The concentration of 22 metals and metalloids after chemical digestion was assessed by inductively coupled plasma-optic emission spectroscopy (ICP-OES) in both the guts and the edible part (EP, muscle + skin) of brown trout. The results, expressed as µg g-1, showed different levels of accumulation in the EP and guts, following the series Cu > Zn > Ba > Al > Sr > Fe > Pb and Fe > Al > Hg > As > Mn > Cu > Ba > B > Zn > Pb, respectively. PCA analysis showed a fairly good correlation between the total lipid and SAFA content and Cd, Hg, and Pb accumulation in the gut. Non-carcinogenic risk assessment, expressed as THQ (target hazard quotient), showed values far below 1 for all metals in muscles, while high As and Hg contamination of the gut draws attention to possible health risks which should be discarded from the fish before consumption. TR (target cancer risk) values showed alarmingly high values for As and Cd when the fish were consumed entirely (gut + EP), while Pb levels were far below the safety levels.

Food and temperature stressors have opposing effects in determining flexible migration decisions in brown trout (Salmo trutta).

With rapid global change, organisms in natural systems are exposed to a multitude of stressors that likely co-occur, with uncertain impacts. We explored individual and cumulative effects of co-occurring environmental stressors on the striking, yet poorly understood, phenomenon of facultative migration. We reared offspring of a brown trout population that naturally demonstrates facultative anadromy (sea migration), under different environmental stressor treatments and measured life history responses in terms of migratory tactics and freshwater maturation rates. Juvenile fish were exposed to reduced food availability, temperatures elevated to 1.8°C above natural conditions or both treatments in combination over 18 months of experimental tank rearing. When considered in isolation, reduced food had negative effects on the size, mass and condition of fish across the experiment. We detected variable effects of warm temperatures (negative effects on size and mass, but positive effect on lipids). When combined with food restriction, temperature effects on these traits were less pronounced, implying antagonistic stressor effects on morphological traits. Stressors combined additively, but had opposing effects on life history tactics: migration increased and maturation rates decreased under low food conditions, whereas the opposite occurred in the warm temperature treatment. Not all fish had expressed maturation or migration tactics by the end of the study, and the frequency of these 'unassigned' fish was higher in food deprivation treatments, but lower in warm treatments. Fish showing migration tactics were smaller and in poorer condition than fish showing maturation tactics, but were similar in size to unassigned fish. We further detected effects of food restriction on hypo-osmoregulatory function of migrants that may influence the fitness benefits of the migratory tactic at sea. We also highlight that responses to multiple stressors may vary depending on the response considered. Collectively, our results indicate contrasting effects of environmental stressors on life history trajectories in a facultatively migratory species.

Immunomodulatory function of antimicrobial peptide EC-Hepcidin1 modulates the induction of inflammatory gene expression in primary cells of Caspian Trout (Salmo trutta caspius Kessler, 1877).

Hepcidins, a group of antimicrobial peptides (AMPs), play a key role in the innate immune system of fishes and act against different pathogens. In this study, antimicrobial and immune-inflammatory activity of a synthetic EC-hepcidin1, previously identified from orange-spotted grouper, were evaluated. EC-hepcidin1 showed weak activity against the zoonotic fish pathogen Streptococcus iniae (MIC 100 µg mL-1 and MBC 150 µg mL-1). To study the effect of AMPs in general, and EC-hepcidin1 in particular, a primary cell culture (SC) from the fin tissue of the Caspian Trout (Salmo trutta caspius) was established. The neutral Red method on SC cells revealed that EC-hepcidin1 has no or very low cytotoxic properties. Treatment of cells with either EC-hepcidin1 (150 µg mL-1) or fish pathogen Streptococcus iniae (MOI = 10) and a mixture of both resulted in the up-regulation of gene expression of MHC-UBA, IL-6, and TNFα indicating the modulatory function on inflammatory processes. These findings indicate that EC-hepcidin1 might act as a candidate for modulation of the innate immune system in S. iniae-based infection.

Renibacterium salmoninarum and Mycobacterium spp.: two bacterial pathogens present at low levels in wild brown trout (Salmo trutta fario) populations in Austrian rivers.

BACKGROUND: Renibacterium salmoninarum and Mycobacterium sp. are important bacterial pathogens of fish. R. salmoninarum is the causative agent of bacterial kidney disease, a Gram-positive bacterium mostly known for causing chronic infections in salmonid fish, while multiple species belonging to the Mycobacterium genus have been associated with mycobacteriosis in fish as well as in human. The objective of this study was to determine the prevalence of these two bacterial pathogens in populations of wild brown trout (Salmo trutta fario) in four rivers (Kamp, Wulka, Traun and Ybbs) in Austria. RESULTS: A total of 457 kidney samples were examined for both bacterial agents using nested and conventional PCR as well as bacterial cultivation on KDM-2, histological examination and immunohistochemistry. Molecular evidence showed an estimated prevalence level of 0.94% for R. salmoninarum in 2017 while the bacterium could not be detected in 2018 and histology showed signs consistent with a low-level chronic inflammation in the kidney of infected fish. Similarly, no fish were found positive for Mycobacterium in 2017 but in 2018, the prevalence was found to be 37.03% in the Kamp river (4.08% across all rivers). The sequencing data confirmed that these fish carried Mycobacterium sp. although the precise species of Mycobacterium could not be ascertained. CONCLUSIONS: This survey constitutes the first insight into the prevalence rate of R. salmoninarum and Mycobacterium sp. in wild brown trout (Salmo trutta fario) populations in Austria. Both of these pathogens were only detected in the summer months (June and July), which might suggest that the stress linked to increased water temperature could act as stressor factor and contribute to the outbreak of these diseases. The age of the fish might also play a role, especially in the case of Mycobacterium sp. as all the infected fish were in their first summer (June).

Mysterious syndrome causing high mortality in wild brown trout in Eastern Switzerland, pathology and search for a possible cause.

Since 2016, annually occurring species-specific die-offs of brown trout (Salmo trutta fario) occurred in the Thur River, situated in the Eastern part of Switzerland. These events lead to drastically reduced population densities in the impacted river regions. Clinical signs in brown trout and mortality were restricted to few weeks in August/September. To characterize the syndrome and to find possible causes, from end of March to November 2018, one-year-old brown trout (Salmo trutta fario) and rainbow trout (Oncorhynchus mykiss) were exposed to water from Thur River, fish were sampled regularly and screened for infectious agents, including viral metagenomics, and pathology was described. Starting approximately four months post-exposure, brown trout showed severe lymphohistiocytic pancarditis and necrotizing and haemorrhagic hepatitis. These lesions were recorded until the end of the experiment in November. Rainbow trout were not affected at any point in time. No infectious agents could be identified so far as cause of disease, especially no viral aetiology. Even if pathogenesis and pathology point in the direction of an infectious agent, a causative relationship could not be confirmed and aetiology remains unclear.