Brown trout in Oder estuary tributaries: genetic structure, stocking, and admixture.

Several rivers that are tributaries of the Oder estuary are inhabited by Salmo trutta L, the most important of which are Ina, Gowienica, and Wolczenica. Both forms of the species, sea trout and resident brown trout, are present. All rivers are traditionally stocked with either sea trout from the neighboring Pomeranian river Rega basin or resident brown trout from various locations. To examine populations in these rivers in terms of genetic structure, genetic diversity, and origin, they were analyzed using 13 microsatellite loci. Relatedness was also assessed for fish stocked in the same year. The obtained genotypes were compared with breeding stocks used for stocking in Poland. The analyses revealed a significant genetic distance between adult individuals from Ina and Rega Rivers and fish caught during electrofishing. Strong kinship relationships were identified in the sampled areas, with high proportions of fish originating from stocking and their dominance in numbers over wild juveniles, primarily in smaller tributaries. Additionally, clear separation in the origin of stocked individuals was observed. Adult trout from Ina and Rega are genetically closer to northern brown trout lineages, providing crucial information for the management and biodiversity conservation of Polish Salmo trutta populations.

Leveraging the genetic diversity of trout in the rivers of the British Isles and northern France to understand the movements of sea trout (Salmo trutta L.) around the English Channel.

Populations of anadromous brown trout, also known as sea trout, have suffered recent marked declines in abundance due to multiple factors, including climate change and human activities. While much is known about their freshwater phase, less is known about the species' marine feeding migrations. This situation is hindering the effective management and conservation of anadromous trout in the marine environment. Using a panel of 95 single nucleotide polymorphism markers we developed a genetic baseline, which demonstrated strong regional structuring of genetic diversity in trout populations around the English Channel and adjacent waters. Extensive baseline testing showed this structuring allowed high-confidence assignment of known-origin individuals to region of origin. This study presents new data on the movements of anadromous trout in the English Channel and southern North Sea. Assignment of anadromous trout sampled from 12 marine and estuarine localities highlighted contrasting results for these areas. The majority of these fisheries are composed predominately of stocks local to the sampling location. However, there were multiple cases of long-distance movements of anadromous trout, with several individuals originating from rivers in northeast England being caught in the English Channel and southern North Sea, in some cases more than 1000 km from their natal region. These results have implications for the management of sea trout in inshore waters around the English Channel and southern North Sea.

Thermotactic behaviour in lacustrine and riverine forms of Salmo trutta and its relevance to an emerging parasitic disease (PKD) in the wake of climate change.

The thermotactic response of brown trout (Salmo trutta) was examined with the goal to investigate potential effects of the emerging temperature-dependent fatal trout disease PKD (proliferative kidney disease). First the differences in cold-water preferences of two forms of brown trout, lacustrine (migratory) and riverine, were determined. Second, it was studied whether this preference was changed in fish infected with PKD. The experiment involved a one-week habituation period at 14 °C in a two-chamber runway followed by a week of 3 °C temperature difference between the two runways. The fish could freely move between lanes via an opening at the end where food was provided. The temperature manipulation was repeated twice, and there were 3 trials per experimental group. All fish developed a clear spatial preference in the test. Lacustrine trout demonstrated a preference for warmer water, while riverine trout preferred cooler water. This may increase the risk to PKD in the lacustrine form. Most strikingly, riverine trout experimentally exposed to Tetracapsuloides bryosalmonae, the parasite that causes PKD, demonstrated stronger cold-seeking behaviour than control fish. Cold seeking behaviour suggests the occurrence of a disease-induced behavioural chill response, which may play an important role in disease recovery. This demonstrates the significance of protecting river connectivity and cold-water sanctuaries as management strategies for preserving salmonid populations in a warming climate.

Interrelation between environmental conditions, acanthocephalan infection and metal(loid) accumulation in fish intestine: an in-depth study.

Metal(loid) bioaccumulation in acanthocephalans (Dentitruncus truttae) and intestines of fish (Salmo trutta) from the Krka River, influenced by industrial and municipal wastewaters, was investigated in relation to exposure to metal(loid)s from fish gut content (GC), water, and sediment to estimate potentially available metal (loid)s responsible for toxic effects and cellular disturbances in biota. Sampling was performed in two seasons (spring and autumn) at the reference site (river source, KRS), downstream of the wastewater outlets (Town of Knin, KRK), and in the national park (KNP). Metal(loid) concentrations were measured by ICP-MS. The highest accumulation of As, Ba, Ca, Cu, Fe, Pb, Se and Zn was observed mainly in organisms from KRK, of Cd, Cs, Rb and Tl at KRS, and of Hg, Mn, Mo, Sr and V at KNP. Acanthocephalans showed significantly higher bioaccumulation than fish intestine, especially of toxic metals (Pb, Cd and Tl). Metal(loid) bioaccumulation in organisms partially coincided to exposure from water, sediments and food, while in GC almost all elements were elevated at KNP, reflecting the metal(loid) exposure from sediments. Seasonal differences in organisms and GC indicated higher metal (loid) accumulation in spring, which follows enhanced fish feeding rates. Higher number of acanthocephalans in the intestine influenced biodilution process and lower concentrations of metal(loid)s in fish, indicating positive effects of parasites to their host, as supported by high values of bioconcentration factors. Fish intestine and acanthocephalan D. truttae were confirmed as sensitive indicators of available metal fraction in conditions of generally low environmental exposure in karst ecosystem. Since metal(loid) accumulation depended on ecological, chemical and biological conditions, but also on the dietary habits, physiology of organisms and parasite infection, continuous monitoring is recommended to distinguish between the effects of these factors and environmental exposure when assessing dietary associated metal(loid) exposure in aquatic organisms.

A Proof-of-Concept for a Hypolipidemic Brown Trout Model.

The impacts of hypolipidemic pharmaceuticals on fish lipid metabolism remain unexplored. However, data points to similar effects and mechanisms of action between fish and humans. Therefore, fish may be a strong model for screening hypolipidemic drug candidates and water pollution by lipid-modulating agents. This study aimed to test a new hypolipidemic model assay with juvenile brown trout using atorvastatin (ATV)-a hypolipidemic chemical. We selected 17α-ethinylestradiol (EE2), known to cause hyperlipidemia in fish, to ensure model functionality. Fish received intramuscular injections of 4 µL/g for two weeks under the following experimental conditions: control-C (0.7% NaCl), solvent control-SC (0.7% NaCl, 0.9% ethanol, 0.1% dimethyl sulfoxide), ATV (0.3 µg/g), EE2 (2 µg/g), and a mixture of both compounds-MIX (0.3 µg/g ATV and 2 µg/g EE2). Endpoints included blood lipid biochemistry, hepatic lipid droplet quantification, and liver mRNA expression of lipid-related target genes (related to lipogenesis, lipid transport, and ß-oxidation pathways). ATV lowered blood total cholesterol, high-density lipoproteins (HDL), and low-density lipoproteins (LDL) levels, whilst triglycerides and very-low-density lipoproteins (VLDL) were highest under EE2. Hepatic lipid droplet deposition significantly increased in the ATV, EE2, and MIX groups. ATV and MIX caused a significant downregulation of the peroxisome proliferator-activated receptor γ (pparγ) and acetyl Co-A oxidase 3 (acox3). EE2 upregulated acyl-CoA long-chain synthetase 1 (acsl1) and downregulated both fatty acid binding protein 1 (fabp1) and acetyl Co-A oxidase 1-3I (acox1-3I). ATV caused hypolipidemic effects in juvenile brown trout and could even counteract EE2-stimulated hyperlipidemia, reinforcing the potential of fish hypo- and hyperlipidemic models.

Resource instability undermines predictable plasticity-mediated morphological responses to diet in a postglacial fish.

Phenotypic plasticity has been presented as a potential rapid-response mechanism with which organisms may confront swift environmental change and increasing instability. Among the many difficulties potentially facing freshwater fishes in recently glaciated ecosystems is that of invertebrate prey communities becoming significantly altered in species composition and relative abundance. To test how the rapidity of diet resource change may affect phenotypic responses during development, we subjected juvenile brown trout to pelagic-type or littoral-type diets that alternated either daily, sub-seasonally, or not at all over a single growth season. The proportional intake of each diet was traced with stable isotopes of carbon and nitrogen and modelled with morphometric data on head and jaw shape. While those trout exposed to a single diet type developed predictable morphologies associated with pelagic or littoral foragers, those raised on alternating diets expressed more unpredictable morphologies. With extreme (daily) or even sub-seasonal (monthly) resource instability, the association of diet type with the phenotype was overwhelmed, calling into question the efficacy of plasticity as a means of adaptation to environments with rapidly fluctuating prey resources.

Interspecific competitive interactions affect body size and oxidative status of two nonnative salmonid species.

In fish, interspecific interactions between nonnative and other sympatric species are considered determinants in shaping species assemblages. Such interactions can also arise between nonnative fish species only, including salmonids such as the brown trout (Salmo trutta, Linnaeus, 1758) and the rainbow trout (Oncorhynchus mykiss, Walbaum, 1792), returning contrasting outcomes. The present manipulative experiment was aimed at exploring the effect of interspecific competition on the body growth and the oxidative status of parr (2 + -year-old individuals) of the brown trout and the rainbow trout. Allopatric (intraspecific competition) and sympatric (interspecific competition) populations of these species were experimentally recreated in two wild streams. At the end of a 2-month-long experiment, changes in specific growth rate (SGR), oxidative status (i.e., levels of reactive oxygen species and activity of antioxidant enzymes such as superoxide dismutase - SOD, catalase - CAT and glutathione peroxidase - GPx) and oxidative damage (i.e., lipid peroxidation) were investigated in brown and rainbow trout individuals maintained in allopatric or sympatric populations. Sympatric interactions between rainbow and brown trout parr resulted in a significant decrease in SGR of brown trout individuals only. Moreover, an overall modulation of the oxidative status, in terms of an increase in ROS levels coupled with the activation of SOD and CAT activity, occurred in brown trout individuals under sympatric conditions. These findings might suggest that, under sympatric conditions, parr of the rainbow trout are more competitive than brown trout for food acquisition. However, this competition affected the antioxidant defenses of the brown trout only, probably because of reduced ingestion of dietary antioxidants or increased physical activity and aggressive behavior. Thus, interspecific interactions can induce physiological and phenotypic effects on parr of nonnative salmonids, with potential consequences on the establishment of populations of these species in freshwater ecosystems.

Distinct dynamics in mountain watersheds: Exploring mercury and microplastic pollution-Unraveling the influence of atmospheric deposition, human activities, and hydrology.

The intensification of human activities all around the globe has led to the spread of micropollutants in high-mountain freshwater environments. We therefore aimed to assess the geospatial distribution and determine the potential sources of (total-) mercury (THg) and microplastics (MPs) in mountain freshwater ecosystems. To do so, we analyzed THg and MP concentrations in brown trout, biofilm, and sediments from lotic and lentic ecosystems in the Pyrenees - all subjected to different types of human pressure. Additionally, we assessed the potential impacts of these pollutants on fish, and explored the bioindication capacity of brown trout (Salmo trutta fario) and biofilm regarding THg and MP pollution. For the first time, we measured concentrations of MPs trapped in the matrix of freshwater biofilm. Our results suggest that THg in the Pyrenees might be explained by both legacy (regional) and distant sources, in combination with environmental characteristics such as the presence of peatlands or streamwater physicochemistry, while MPs in fish are linked to recent local pollution sources such as single-use plastics. In contrast, MPs in biofilm matrix and sediments indicate a combination of distant (i.e., atmospheric deposition) and recent local pollution sources. Moreover, hydrodynamics and plastic density likely control MP distribution in rivers. Based on Fulton's condition factor, we also found that higher THg concentrations caused a negative impact on fish health (K < 1), while no impact of MPs could be seen. Therefore, we suggest that brown trout and biofilm can serve as bioindicators of atmospheric deposition of THg in high-altitude lakes and that biofilm is a reliable bioindicator to assess MP pollution in remote environments. Brown trout may also act as a bioindicator of MP pollution, but only efficiently in more polluted areas.

Liver and Plasma Fatty Acid Characterization in Cultured Brown Trout at Distinct Reproductive Stages.

Fatty acids are energy sources, and their profiles are used as biomarkers of metabolic status and physiological changes in fish. Within this context, the main aim of this study was to identify the fatty acids that best discriminate the reproductive status of male and female farmed brown trout. The fatty acid composition in liver and plasma samples from the adults of both sexes was monitored along four distinct reproductive stages, namely the spawning capable (December), regressing (March), regenerating (July), and developing (November) stages. Irrespective of the sex and stage, the most representative fatty acids were palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1 n-9), arachidonic acid (20:4 n-6), eicosapentaenoic acid (EPA, 20:5 n-3), and docosahexaenoic acid (DHA, 22:6 n-3). There were no significant sex differences in fatty acid classes in the liver and plasma. Despite this, there were several changes in individual fatty acid levels between the sexes. In the liver, both males and females showed high monounsaturated fatty acid and low polyunsaturated fatty acid (PUFA) levels during the regressing and regenerating stages. At spawning capable and developing stages, a reverse profile was noted. The plasma profiles were mainly influenced by changes in saturated fatty acids and PUFAs in males and by PUFA in females. Based on the most representative fatty acids, four patterns were established for female plasma samples, one for each reproductive stage. This scenario suggests that female plasma samples are promising for the discrimination of gonadal reproductive status, and this potential can be further explored in aquaculture and environmental monitoring studies.

Behaviour of anadromous brown trout (Salmo trutta) in a hydropower regulated freshwater system.

Many Norwegian rivers and lakes are regulated for hydropower, which affects freshwater ecosystems and anadromous fish species, such as sea trout (Salmo trutta). Lakes are an important feature of many anadromous river systems. However, there is limited knowledge on the importance of lakes as habitat for sea trout and how hydropower affects the behaviour of sea trout in lakes. To investigate this, we conducted an acoustic telemetry study. A total of 31 adult sea trout (532 ± 93 mm total length) were captured by angling in river Aurlandselva, Norway, and tagged between July 20 and August 12, 2021. The tags were instrumented with accelerometer, temperature, and depth sensors, which provided information on the sea trout's presence and behaviour in lake Vassbygdevatnet. Our results indicate that there was a large prevalence of sea trout in the lake during the spawning migration, and that the sea trout were less active in the lake compared to the riverine habitats. An increase in activity of sea trout in the lake during autumn might indicate that sea trout spawn in the lake. However, the discharge from the high-head storage plant into the lake did not affect the depth use or activity of sea trout in the lake. Furthermore, the large prevalence of spawners in the lake during autumn will likely cause an underestimation of the size of the sea trout population in rivers with lakes during annual stock assessment. In conclusion, our results could not find evidence of a large impact of the discharge on the behaviour of sea trout in the lake.

Do Organochlorine Contaminants Modulate the Parasitic Infection Degree in Mediterranean Trout (Salmo trutta)?

We investigated the occurrence of organochlorine pollutants (OCs) in the muscle of brown trout and evaluated their potential modulation of parasite infection. The toxicological risk for consumer health was assessed, too. Trout were collected from the Sila National Park (Calabria region, South of Italy). The highest concentrations emerged for the sum of the 6 non-dioxin-like (ndl) indicator polychlorinated biphenyls (Σ6ndl-PCBs), followed by the 1,1,1-trichloro-2,2-di(4-chlorophenyl)-ethane (DDT), dioxin-like PCBs, hexachlorobenzene (HCB), and dieldrin. Measured on lipid weight (LW), the mean value of Σ6ndl-PCBs amounted to 201.9 ng g-1, that of ΣDDTs (the sum of DDT-related compounds) to 100.2 ng g-1, with the major contribution of the DDT-metabolite p,p'-DDE which was detected in all sample units (97.6 ng g-1 on average). Among dioxin-like congeners, PCB 118 showed the highest mean concentration (21.96 ng g-1 LW) and was detected in all sample units. Regression analysis of intestinal parasites on OC concentration was performed, controlling for two potential confounding factors, namely sex and sexual stage. The results evidenced the existence of interactions between the dual stressors in the host-parasite system in the wild. A negative and statistically significant correlation was estimated, suggesting that OCs may decrease parasite infection degree. Regarding the toxicological risk evaluation, OC concentrations were consistently below the current European Maximum Residue Limits.

Genetic structure and relatedness of brown trout (Salmo trutta) populations in the drainage basin of the Ölfusá river, South-Western Iceland.

Background: Lake Þingvallavatn in Iceland, a part of the river Ölfusá drainage basin, was presumably populated by brown trout soon after it formed at the end of the last Ice Age. The genetic relatedness of the brown trout in Þingvallavatn to other populations in the Ölfusá drainage basin is unknown. After the building of a dam at the outlet of the lake in 1959 brown trout catches declined, though numbers have now increased. The aim of this study was to assess effects of geographic isolation and potential downstream gene flow on the genetic structure and diversity in brown trout sampled in several locations in the western side of the watershed of River Ölfusá. We hypothesized that brown trout in Lake Þingvallavatn constituted several local spawning populations connected by occasional gene flow before the damming of the lake. We also estimated the effective population size (NE) of some of these populations and tested for signs of a recent population bottleneck in Lake Þingvallavatn. Methods: We sampled brown trout inhabiting four lakes and 12 rivers within and near the watershed of River Ölfusá by means of electro- and net- fishing. After stringent data filtering, 2,597 polymorphic loci obtained from ddRADseq data from 317 individuals were ascertained as putative neutral markers. Results: Overall, the genetic relatedness of brown trout in the Ölfusá watershed reflected the connectivity and topography of the waterways. Ancestry proportion analyses and a phylogenetic tree revealed seven distinct clusters, some of which corresponded to small populations with reduced genetic diversity. There was no evidence of downstream gene flow from Lake Þingvallavatn, although gene flow was observed from much smaller mountain populations. Most locations showed low NE values (i.e., ~14.6 on average) while the putative anadromous trout from River Sog and the spawning population from River Öxará, that flows into Lake Þingvallavatn, showed notably higher NE values (i.e., 71.2 and 56.5, respectively). No signals of recent population bottlenecks were detected in the brown trout of Lake Þingvallavatn. Discussion: This is the first time that the genetic structure and diversity of brown trout in the watershed of River Ölfusá have been assessed. Our results point towards the presence of a metapopulation in the watershed of Lake Þingvallavatn, which has been influenced by restoration efforts and is now dominated by a genetic component originated in River Öxará. Many of the locations studied represent different populations. Those that are isolated in headwater streams and lakes are genetically distinct presenting low genetic diversity, yet they can be important in increasing the genetic variation in downstream populations. These populations should be considered for conservation and direct management.

Modifying and parameterizing the individual-based model inSTREAM for Atlantic salmon and brown trout in the regulated Gullspång River, Sweden.

We modified, parameterized, and applied the individual-based model inSTREAM version 6.1 for lake-migrating populations of landlocked Atlantic salmon (Salmo salar) and brown trout (S. trutta) in a residual flow stretch of the hydropower-regulated Gullspång River, Sweden. This model description is structured according to the TRACE model description framework. Our aim was to model responses in salmonid recruitment to alternative scenarios of flow release and other environmental alterations. The main response variable was the number of large out-migrating juvenile fish per year, with the assumption that individuals are more inclined to out-migrate the larger they get, and that migration is an obligatory strategy. Population and species-specific parameters were set based on local electrofishing surveys, redd surveys, physical habitat surveys, broodstock data as well as scientific literature.•Simulations were set to run over 10 years, with sub-daily time steps, in this spatially and temporally explicit model.•Model calibration and validation of fish growth was done using data on juvenile fish from electrofishing.•The results were found to be sensitive to parameter values for aggregated fish, i.e., "superindividuals" and for the high temperature limit to spawning.

A Step Forward in the Characterization of Primary Brown Trout Hepatocytic Spheroids as Experimental Models.

Mammal hepatocyte spheroids have been investigated as alternative experimental models in several contexts, since three-dimensional (3D) systems have shown the potential to mimic in vivo scenarios. The description of fish hepatocyte 3D models is still minimal. This study intends to further characterize brown trout primary hepatocyte spheroids at distinct time points up to 25 days in culture. Viability, biometry, histomorphology, and basal expression of a selection of genes (metabolism and detoxification, efflux transport, and estrogenic signalling) were considered. The gene expression of whole liver samples from the same fish donor were evaluated concurrently. After 12 days in culture, the hepatocyte spheroids exhibited biometric and morphological stability. From the 12th to the 20th day in culture, the basal expression levels for most of the selected genes did not vary. The targeted mRNA levels were higher in brown trout liver samples compared to hepatocyte spheroids. Despite that, data supported that this model resembles some in vivo features. As an experimental alternative model, it showed potential to be used in a stable time window that can be exploited for exposure tests to different xenobiotics, namely, estrogenic compounds.

Metal Contaminants in Fish: Blood as a Potential Non-lethal Monitoring Tool.

The use of fish to monitor metal contamination is well established, but existing studies often focus on internal tissues that require the sacrifice of organisms. Developing non-lethal methods is thus a scientific challenge to enable large scale biomonitoring of wildlife health. We explored blood as a potential non-lethal monitoring tool for metal contamination in brown trout (Salmo trutta fario) as a model species. First, we investigated differences in metal contamination loads (i.e., Cr, Cu, Se, Zn, As, Cd, Pb and Sb) in different blood components (whole blood, red blood cells and plasma). Whole blood was reliable to measure most metals, implying that blood centrifugation is not necessary, thus minimizing sample preparation time. Second, we measured the within individual distribution of metals across tissues (whole blood, muscle, liver, bile, kidney and gonads) to test if blood could be a reliable monitoring tool compared to other tissues. Results show that the whole blood was reliable compared to muscle and bile to measure the levels of metals such as Cr, Cu, Se, Zn, Cd and Pb. This study opens the possibility for future ecotoxicological studies in fish to use blood instead of internal tissues to quantify some metals, thus reducing the negative impacts of biomonitoring on wildlife.

Genetic structure of Ponto-Caspian trout populations shows gene flow among river drainages and supports resident Salmo rizeensis as a genetically distinct taxon.

To assess the genetic structure of Ponto-Caspian brown trout (Salmo trutta complex) populations, we analyzed both mitochondrial DNA sequences and genotypes at 10 microsatellite loci of fish caught in the Black Sea and from nine river catchments in Georgia, flowing into either the Black or Caspian seas. The results show that: (1) there is substantial genetic differentiation among Ponto-Caspian trout populations, both among the populations of different nominal species and within those of the same species; (2) the genetic distance between conspecific populations from the Black and Caspian Sea basins exceeds that among the populations within the same basin. Moreover, within drainages, genetic distance correlates with the geographic distance; (3) the Black Sea itself is not a barrier to gene flow among the watersheds draining into the Black Sea; (4) some populations in the headwaters of the rivers draining into the Black Sea Basin fall out of this pattern and likely form a separate, non-anadromous (resident) taxon, previously described from northeastern Turkey as Salmo rizeensis. This hypothesis is supported by mitochondrial DNA phylogeny. The presence of both anadromous and resident populations in a single river basin calls for a substantial re-thinking of speciation patterns and taxonomy of Eurasian brown trout.

Spatial and intra-host distribution of myxozoan parasite Tetracapsuloides bryosalmonae among Baltic sea trout (Salmo trutta).

Proliferative kidney disease caused by the myxozoan parasite Tetracapsuloides bryosalmonae has been actively studied in juvenile salmonids for decades. However, very little is known about parasite prevalence and its geographical and intra-host distribution at older life stages. We screened T. bryosalmonae among adult sea trout (Salmo trutta) (n = 295) collected along the Estonian Baltic Sea coastline together with juvenile trout from 33 coastal rivers (n = 1752) to assess spatial infection patterns of the adult and juvenile fish. The parasite was detected among 38.6% of adult sea trout with the prevalence increasing from west to east, and south to north, along the coastline. A similar pattern was observed in juvenile trout. Infected sea trout were also older than uninfected fish and the parasite was detected in sea trout up to the age of 6 years. Analysis of intra-host distribution of the parasite and strontium to calcium ratios from the otoliths revealed that (re)infection through freshwater migration may occur among adult sea trout. The results of this study indicate that T. bryosalmonae can persist in a brackish water environment for several years and that returning sea trout spawners most likely contribute to the parasite life cycle by transmitting infective spores.

Absence of Staphylococcus aureus in Wild Populations of Fish Supports a Spillover Hypothesis.

Staphylococcus aureus is a human commensal and opportunistic pathogen that also infects other animals. In humans and livestock, where S. aureus is most studied, strains are specialized for different host species. Recent studies have also found S. aureus in diverse wild animals. However, it remains unclear whether these isolates are also specialized for their hosts or whether their presence is due to repeated spillovers from source populations. This study focuses on S. aureus in fish, testing the spillover hypothesis in two ways. First, we examined 12 S. aureus isolates obtained from the internal and external organs of a farmed fish. While all isolates were from clonal complex 45, genomic diversity indicates repeated acquisition. The presence of a φSa3 prophage containing human immune evasion genes suggests that the source was originally human. Second, we tested for S. aureus in wild fish that were isolated from likely sources. In particular, we sampled 123 brown trout and their environment at 16 sites in the remote Scottish Highlands with variable levels of exposure to humans, birds, and livestock. This screen found no S. aureus infection in any of the wild populations or their environment. Together, these results support that the presence of S. aureus in fish and aquaculture is due to spillover from humans rather than specialization. Given the trends of increasing fish consumption, a better understanding of the dynamics of S. aureus spillover in aquaculture will mitigate future risks to fish and human health. IMPORTANCE Staphylococcus aureus is a human and livestock commensal but also an important pathogen responsible for high human mortality rates and economic losses in farming. Recent studies show that S. aureus is common in wild animals, including fish. However, we do not know whether these animals are part of the normal host range of S. aureus or whether infection is due to repeated spillover events from true S. aureus hosts. Answering this question has implications for public health and conservation. We find support for the spillover hypothesis by combining genome sequencing of S. aureus isolates from farmed fish and screens for S. aureus in isolated wild populations. The results imply that fish are unlikely to be a source of novel emergent S. aureus strains but highlight the prominence of the spillover of antibiotic-resistant bacteria from humans and livestock. This may affect both future fish disease potential and the risk of human food poisoning.

Differentially expressed transcripts of Tetracapsuloides bryosalmonae (Cnidaria) between carrier and dead-end hosts involved in key biological processes: novel insights from a coupled approach of FACS and RNA sequencing.

Tetracapsuloides bryosalmonae is a malacosporean endoparasite that infects a wide range of salmonids and causes proliferative kidney disease (PKD). Brown trout serves as a carrier host whereas rainbow trout represents a dead-end host. We thus asked if the parasite adapts to the different hosts by changing molecular mechanisms. We used fluorescent activated cell sorting (FACS) to isolate parasites from the kidney of brown trout and rainbow trout following experimental infection with T. bryosalmonae. The sorted parasite cells were then subjected to RNA sequencing. By this approach, we identified 1120 parasite transcripts that were expressed differentially in parasites derived from brown trout and rainbow trout. We found elevated levels of transcripts related to cytoskeleton organisation, cell polarity, peptidyl-serine phosphorylation in parasites sorted from brown trout. In contrast, transcripts related to translation, ribonucleoprotein complex biogenesis and subunit organisation, non-membrane bounded organelle assembly, regulation of protein catabolic process and protein refolding were upregulated in rainbow trout-derived parasites. These findings show distinct molecular adaptations of parasites, which may underlie their distinct outcomes in the two hosts. Moreover, the identification of these differentially expressed transcripts may enable the identification of novel drug targets that may be exploited as treatment against T. bryosalmonae. We here also describe for the first time how FACS based isolation of T. bryosalmonae cells from infected kidney of fish fosters research and allows to define differentially expressed parasite transcripts in carrier and dead-end fish hosts.

Importance of artificial high flows in maintaining the ecological integrity of a regulated river.

Artificial high flows attempt to simulate natural flood pulses in flow-regulated rivers with the intent to improve their ecological integrity. The long-term use of such high flow events have shown beneficial ecological effects on various rivers globally. However, such responses are often non-linear and characterized by underlying feedback mechanisms among ecosystem components. The question arises as to what happens when such high flow releases are disrupted or even discontinued. Here, we used the long-term (22 years) monitoring dataset from the river Spöl to examine whether discontinuation (2016-2021) of the flood program (annual artificial high flows from 2000 to 2016) resulted in the ecological degradation of the river. We used monitoring data of physico-chemistry, periphyton, benthic organic matter, macroinvertebrates and fish (brown trout, Salmo trutta fario L.) in the analysis. The flood program had no long-term effect on water physico-chemistry with most parameters showing typical variations associated with season and inter-annual weather patterns. The floods were effective at mobilizing bed sediments that reduced periphyton biomass and benthic organic matter following each flood. Increases in periphyton biomass and benthic organic matter occurred between floods, but both parameters showed no significant increase with discontinuation of the flood program. Floods reduced macroinvertebrate densities, but with density increases occurring between floods. The pulsed disturbances, and the progressive change in the habitat template, resulted in shifts in community assembly by reducing densities of Gammarus fossarum, a dominant crustacean, which allowed other taxa to colonize the system. Macroinvertebrate densities remained low after discontinuation of the floods, although G. fossarum densities have increased substantially while other taxa, especially some stoneflies, remained low in abundance. Notably, community assembly returned to a pre-flood composition with discontinuation of the floods. The abundance of brown trout increased substantially during the flood program but returned to low pre-flood numbers with discontinuation of the floods. We conclude that the flood program was beneficial to the ecology of the river Spöl and discontinuation of the floods resulted in degradation of the system after a relatively short lag period. However, the system showed high resilience to an earlier perturbation, a sediment spill in 2013, suggesting a rapid positive response by biota with resumption of the flood program.