Genetic diversity and phylogenetic origin of brown trout Salmo trutta populations in eastern Balkans

The study focuses on the phylogenetic origin and genetic diversity of brown trout in the eastern part of the Balkan Peninsula. It further aims to reveal the impact of human-mediated transfers and stocking with non-indigenous trout on the populations in this area. For these purposes, mtDNA control region and microsatellite variation of 204 individuals from 16 populations were analysed. The results indicate that mtDNA haplotypes from the lower Danube basin and southern Black Sea basins differ substantially from a subclade of the Danubian lineage consisting of haplotypes found so far in the most of the Danube basin and in the Caspian and Aral Sea basins. Considering also the results of demographic analyses, this study evidences a complex evolutionary history of brown trout in the southern and western parts of the Black Sea basin. In the Aegean Sea basin, a high frequency of the central haplotype of Adriatic mtDNA lineage has been found. The other Adriatic lineage haplotypes found in this basin differ from the central haplotype by one mutational step only, indicating a recent evolution of the Adriatic lineage in the Aegean Sea basin. Substantial genetic differentiation among populations and basins was revealed. The hybridization with Atlantic brown trout was indicated in both sea basins, but especially in the Danube basin. Compared to other European regions, it can be inferred that the introgression of exogenous brown trout in the eastern Balkan populations is rather low.     

Demographic Genetics of Brown Trout (Salmo Trutta) and Estimation of Effective Population Size from Temporal Change of Allele Frequencies

We studied temporal allele frequency shifts over 15 years and estimated the genetically effective size of four natural populations of brown trout (Salmo trutta L.) on the basis of the variation at 14 polymorphic allozyme loci. The allele frequency differences between consecutive cohorts were significant in all four populations. There were no indications of natural selection, and we conclude that random genetic drift is the most likely cause of temporal allele frequency shifts at the loci examined. Effective population sizes were estimated from observed allele frequency shifts among cohorts, taking into consideration the demographic characteristics of each population. The estimated effective sizes of the four populations range from 52 to 480 individuals, and we conclude that the effective size of natural brown trout populations may differ considerably among lakes that are similar in size and other apparent characteristics. In spite of their different effective sizes all four populations have similar levels of genetic variation (average heterozygosity) indicating that excessive loss of genetic variability has been retarded, most likely because of gene flow among neighboring populations.     

Fine-spotted brown trout: genetic aspects and the need for conservation

In Lake Svartavasstjønn on the Hardangervidda plateau, a fine-spotted variant of brown trout has been found. The spotted variability shows a classical Mendelian mode of inheritance. In this population genetic variability was found in LDH-5 *, MDH-3 * and MDH-3 *. The fast allele at LDH-5 * had a frequency of 0,913. This is remarkable, as this allele has been found in frequencies above 0.4 in only five out of 50 investigated populations in Norway. The genetic characteristics of brown trout from Lake Svartavasstjønn were compared with 11 other populations sampled from a broad geographical area of Norway. Among these was Lake Setningen, where fine-spotted individuals occur at low frequency. Using a cluster analysis based on eight loci found to be polymorphic in brown trout, we found no indication of a close relationship between the fine-spotted brown trout from Lake Svartavasstjønn and the population in Lake Setningen. During a field survey in 1990 we found that the reproduction of fine-spotted trout in Lake Svartavasstjønn had terminated. The fine-spotted trout represents a valuable but threatened genetic resource, and attempts are carried out to conserve it.     

Coping with divided attention: the advantage of familiarity

The ability of an animal to perform a task successfully is limited by the amount of attention being simultaneously focused on other activities. One way in which individuals might reduce the cost of divided attention is by preferentially focusing on the most beneficial tasks. In territorial animals where aggression is lower among familiar individuals, the decision to associate preferentially with familiar conspecifics may therefore confer advantages by allowing attention to be switched from aggression to predator vigilance and feeding. Wild juvenile brown trout were used to test the prediction that familiar fishes respond more quickly than unfamiliar fishes to a simulated predator attack. Our results confirm this prediction by demonstrating that familiar trout respond 14% faster than unfamiliar individuals to a predator attack. The results also show that familiar fishes consume a greater number of food items, foraging at more than twice the rate of unfamiliar conspecifics. To the best of our knowledge, these results provide the first evidence that familiarity-biased association confers advantages through the immediate fitness benefits afforded by faster predator-evasion responses and the long-term benefits provided by increased feeding opportunities.     

Genetic introgression between wild and stocked brown trout in the Douro River basin, Spain

The genetic diversity of Spanish brown trout is currently threatened by stocking with exogenous brown trout from Central and Northern Europe. In the Douro River basin 25% of the analysed populations in the present study showed introgression by genes of hatchery origin. The mean introgression estimated by the single locus approach ( S ) varied from 0 to 22% among populations, with a mean value of 3%. The hatchery allele markers were absent in populations where stocking ceased in 1993. However, the introgression effect was observed in all populations stocked until 1998. It seems that cessation of stocking is a good measure for restoring native populations. A thorough review of published and present data of genetic interactions between wild and stocked brown trout in Spanish rivers indicates different levels of introgression between basins. The absence of a clear geographical pattern in the introgression level suggests that ecological interactions and local stocking programmes may play an important role in stocking success. Finally, several guidelines are provided for conservation and management of native brown trout populations in Spanish rivers.     

Behavioral and neuroendocrine correlates of displaced aggression in trout

In humans and other primates, violent actions performed by victims of aggression are often directed toward an individual or object that is not the source of provocation. This psychological phenomenon is often called displaced aggression. We demonstrate that displaced aggression is either rooted in evolutionarily conserved behavioral and neuroendocrine mechanisms, or represent a convergent pattern that has arisen independently in fish and mammals. Rainbow trout that briefly encountered large, aggressive fish reacted with increased aggression toward smaller individuals. There was a strong negative correlation between received aggression and behavioral change: Individuals subjected to intense aggression were subdued, while moderate assaults induced strong agitation. Patterns of forebrain serotonin turnover and plasma cortisol suggest that the presence of socially subordinate fish had an inhibitory effect on neuroendocrine stress responses. Thus, subordinate individuals may serve as stress-reducing means of aggressive outlet, and displaced aggression toward such individuals appears to be a behavioral stress coping strategy in fishes.     

Predator avoidance behaviour in wild and hatchery‐reared brown trout: the role of experience and domestication

Juvenile brown trout Salmo trutta from natural populations reacted to the presence of piscivorous brown trout by increasing the use of refuges. In contrast, second‐generation hatchery fish and the offspring of wild fish raised under hatchery conditions were insensitive to predation risk. The diel pattern of activity also differed between wild and hatchery brown trout. Second‐generation hatchery fish were predominantly active during daytime regardless of risk levels. Wild fish, however, showed a shift towards nocturnal activity in the presence of predators. These findings emphasize the potential role of domestication in weakening behavioural defences. They support the idea that the behavioural divergence between wild and domesticated individuals can arise from a process of direct or indirect selection on reduced responsiveness to predation risk, or as a lack of previous experience with predators.     

Enhanced phylogeographic information about Austrian brown trout populations derived from complete mitochondrial control region sequences

Complete DNA sequences of the control region revealed a more fine-scale genetic structuring within and among Austrian brown trout Salmo trutta populations providing the opportunity for gene frequency analyses in the phylogeographic context. Ninety-two individuals (75%) were assigned to nine Danubian haplotypes and 31 individuals (25%) comprised seven Atlantic haplotypes of northern European origin. Three of the Atlantic haplotypes were also found in an Austrian hatchery breeding stock.     

Sea trout Salmo trutta in the subarctic: home-bound but large variation in migratory behaviour between and within populations

Anadromous brown trout (sea trout), Salmo trutta, is currently in decline throughout its range, largely due to anthropogenic stressors in freshwater and marine habitats. Acoustic telmetry was utilized to study the marine migration of sea trout post-smolts from three populations in a relatively pristine subarctic fjord system. While at sea, the sea trout spent a substantial part of their time close to their natal river, preferred near shore over pelagic habitats and were strongly surface oriented. Despite a fidelity towards local areas, the sea trout utilized various parts of the fjord system, with maximum dispersion >30 km and total migration distance >300 km. Almost half of the sea trout (44%) migrated between river outlets, indicating that a metapopulation approach may be appropriate when managing neighbouring sea trout populations at high latitudes. Furthermore, the different populations displayed different migratory behaviours in terms of distance migrated, dispersion from origin and the likelihood of leaving their home area. This variation in migratory behaviour is likely influenced by spatiotemporal differences in habitat quality between sites, indicating that local habitat variations may promote population-specific behavioural responses even in relatively confined fjord systems.     

Stock Transfers in Spanish Brown Trout Populations: A Long-term Assessment

Synopsis Stocking of fish from other populations has been commonly employed for enhancement of wild brown trout, Salmo trutta, populations in north Spain. Young hatchery reared brown trout of central European origin were introduced into some Asturian rivers every year since 1984. Based on variation at the isozyme locus LDH-C1* and at the microsatellite locus BFRO 002, two genetic markers race-specific in Salmo trutta, we detected introgression of foreign genomes into native gene pools in some Spanish trout populations where only pure native individuals were present 10 years ago. We strongly suggest development of alternative management policies for conservation of Spanish natural brown trout populations without endangering the traditional recreational fisheries. Jorge I. Izquierdo, Ana G. F. Castillo: These two authors contributed equally to the article.     

Complex genetic diversity patterns of cryptic,sympatric brown trout (<Emphasis Type="Italic">Salmo trutta</Emphasis>) populations in tiny mountain lakes

Intraspecific genetic variation can have similar effects as species diversity on ecosystem function; understanding such variation is important, particularly for ecological key species. The brown trout plays central roles in many northern freshwater ecosystems, and several cases of sympatric brown trout populations have been detected in freshwater lakes based on apparent morphological differences. In some rare cases, sympatric, genetically distinct populations lacking visible phenotypic differences have been detected based on genetic data alone. Detecting such “cryptic” sympatric populations without prior grouping of individuals based on phenotypic characteristics is more difficult statistically, though. The aim of the present study is to delineate the spatial connectivity of two cryptic, sympatric genetic clusters of brown trout discovered in two interconnected, tiny subarctic Swedish lakes. The structures were detected using allozyme markers, and have been monitored over time. Here, we confirm their existence for almost three decades and report that these cryptic, sympatric populations exhibit very different connectivity patterns to brown trout of nearby lakes. One of the clusters is relatively isolated while the other one shows high genetic similarity to downstream populations. There are indications of different spawning sites as reflected in genetic structuring among parr from different creeks. We used >3000 SNPs on a subsample and find that the SNPs largely confirm the allozyme pattern but give considerably lower F _ST values, and potentially indicate further structuring within populations. This type of complex genetic substructuring over microgeographical scales might be more common than anticipated and needs to be considered in conservation management.     

Antioxidative stress proteins and their gene expression in brown trout (Salmo trutta) from three rivers with different heavy metal levels

Three populations of brown trout (Salmo trutta) exposed to different metal levels in their natural environments, were studied with respect to antioxidants metallothionein (MT), superoxide dismutase (SOD) and catalase (CAT) as well as for corresponding mRNA levels. In addition, mRNA levels were studied for glutathione peroxidase (GPx) and glutathione reductase (GR). The Cd/Zn-exposed trout (Naustebekken River) had higher accumulated levels of Cd, Cu and Zn in gills, and higher levels of MT (both protein and mRNA) in liver and kidney as well as in gills compared to the Cu-exposed trout (Rugla River) and trout from an uncontaminated reference river (Stribekken River). Less MT found in the Cu-exposed trout may increase susceptibility to oxidative stress, but no higher levels of antioxidant mRNAs were found in gills of these trouts. The data indicated that chronic exposures of brown trout to Cd, Zn and/or Cu did not involve maintenance of high activities of SOD and CAT enzymes in gills, although SOD mRNA levels were higher in the Cd/Zn-exposed trout. In livers, mRNA levels of SOD, CAT and GPx were higher in the metal-exposed trout, but in the case of GR this was only seen in kidneys of Cd/Zn-exposed trout. However, both metal-exposed groups had higher activities of SOD enzyme in liver compared to the unexposed reference trout, and CAT activity was found to be higher in kidneys of Cu-exposed trout. The Cu-exposed trout did not seem to rely on MT production to avoid Cu toxicity in gills, but rather by keeping the Cu uptake at a low level. A coordinated expression of different stress genes may also be important in chronic metal exposure. It may be concluded that the observed metal effects relies on acclimation rather than on genetic adaptation in the metal exposed populations.     

Early-age changes in oxidative stress in brown trout,Salmo trutta

Fish are often used as models for studies investigating the ability of xenobiotics to induce oxidative stress, though age or developmental stage of the individuals studied has been given little attention. Oxidative stress in other organisms is associated with aging as well as with periods of rapid growth, which occurs in young brown trout. We measured protein carbonyls, 20S proteosome activity and glutathione (GSH) levels in farmed Salmo trutta in four different age groups from 5 months to 3 years. We found an increase in protein carbonyls and a decrease in 20S proteosome activity in both brain and liver tissues of the fish with increasing size and age. Total GSH levels in liver tissue declined as fish aged and the GSSG:GSH ratio increased. Five month and 1 year old trout were treated with paraquat (PQ) to induce oxidative stress. Five month old fish showed no changes in the measured parameters while 1 year old fish had both an increase in protein carbonylation in liver tissue and a decrease in 20S proteosome activity in brain tissue. These results indicate that oxidative stress biomarkers are affected by age or rapid growth in brown trout, and that individuals of different ages respond differently to oxidative stress induced by PQ.     

Behavioral and respiratory responses to stressors in multiple populations of three-spined sticklebacks that differ in predation pressure

Individual animals of the same species inhabiting environments which differ in the frequency and magnitude of stressors often exhibit different physiological and behavioral responses to stressors. Here, we compare the respiratory response to confinement stress, and behavioral responses to ecologically relevant challenges among sticklebacks from 11 different populations varying in predation pressure. We found that sticklebacks from high predation populations breathed faster in response to confinement stress and were, on an average, more behaviorally responsive to a pike behind glass compared with sticklebacks from low predation populations. These patterns differ from the results of studies on other species, highlighting the need for a conceptual framework to understand the proximate and ultimate factors shaping variable responses to stressors over developmental and evolutionary time. Moreover, physiological and behavioral responses were integrated with each other, both at the individual and population levels. In general, fish that were more aggressive and bold in the presence of a predator breathed faster, independent of body size. These results are consistent with the growing body of evidence that individuals differ in a suite of physiological and behavioral mechanisms for coping with challenges in the environment.     

Genetic diversity of brown trout in central Italy

Genetic diversity was analysed in brown trout Salmo trutta populations living in an area of central Italy using RFLP analysis of two mtDNA segments and of the nuclear locus LDH‐C1 *. The data indicated a genetic structure profoundly altered by repeated stockings with allochthonous material of Atlantic origin. In fact, four and 11 of the haplotypes detected were, respectively, identical or genetically very close to haplotypes found in Danish populations, the putative source of stocked brown trout. Furthermore, the LDH‐C1 * 90 allele, typical of north‐western Europe, was widespread among the samples studied. Nonetheless, four populations are characterized by a high frequency of both putative autochthonous haplotypes and the LDH‐C1 * 100 allele, common in the Mediterranean basin. These populations, sampled in areas where S. trutta is documented historically, might represent a remnant of the species' indigenous biodiversity, showing the scope for improving the management of brown trout in central Italy.     

Divergence in behavioural responses to stress in two strains of rainbow trout (Oncorhynchus mykiss) with contrasting stress responsiveness

The aim of this study was to establish whether two lines of rainbow trout divergent for their plasma cortisol response to a standardized stressor would show consistent differences in their behavioural response to a range of challenging situations. Our results show that the high- and low-responding (HR and LR) lines of rainbow trout did not differ in the aggression shown towards an intruder or in their response to the introduction of a novel object to their home environment. However, there was a difference in behaviour between the two selection lines when they were exposed to two unfamiliar environments. These results suggest that the behaviour of the HR and LR fish differs when they are challenged in unfamiliar environments, while their behaviour does not differ when they are challenged in their home environment. These observations are in agreement with studies on mammals that show that individuals with reactive coping styles perform similarly to proactive animals when they are challenged in a familiar environment, while they show different behaviour when they are challenged in unfamiliar environments. Thus, these results provide further evidence that the HR and LR selection lines of rainbow trout exemplify the two different coping styles described in mammals.     

Does individual variation in stress responses and agonistic behavior reflect divergent stress coping strategies in juvenile rainbow trout?

Individual rainbow trout were transferred to visual isolation in experimental aquaria. As a measure of the speed of acclimation, individual food intake was quantified during the first 6 d following transfer. Following acclimation, aggression was quantified by subjecting the fish to three resident-intruder tests, with 30 d of recovery between the tests. Moreover, between the resident-intruder tests (i.e., two times) the fish were exposed to an unfamiliar environment and their cortisol response was measured. The results of this study show that individuals of juvenile rainbow trout differ distinctly in their response to changes in their environment, and that this diversity in behavior is reflected by consistent behavioral traits displayed by individual fish. These traits have proven to be consistent not only over time but also across situations, revealing two distinct behavioral profiles, in the same manner as shown in studies on proactive and reactive mammals. Our results also show that the reactivity of the hypothalamic-pituitary-interrenal (HPI) axis, when exposed to a stressor, is a consistent physiological trait in juvenile rainbow trout. We found that difference in HPI axis reactivity is linked to the different behavioral profiles. However, HPI axis reactivity could not be linked directly to the singular behavioral traits measured. In other words, we did not find that the consistent behavioral traits shown by the fish were associated with a difference in HPI axis reactivity in the same manner as the reactivity of the hypothalamic-pituitary-adrenocortical axis does in mammals. Taken together, our results show that stress coping strategies akin to what has been described as reactive and proactive stress coping in mammals appear to exist in juvenile rainbow trout.     

Competition between brown trout and Atlantic salmon parr over pool refuges during rapid dewatering

Variations in distributions and behaviours of Atlantic salmon Salmo salar in allopatry (homogeneous) and in sympatry with brown trout Salmo trutta (mixed) were observed before, during and after 2 day periods of dewatering in a large glass-sided indoor stream at densities typical of Scottish upland streams. Brown trout utilized pools more than Atlantic salmon at normal flows and in both species the majority of fishes moved into pools during dewatering. There was no significant effect of brown trout, which was the more dominant species, on the overall ability of Atlantic salmon to use pool habitat as a refuge during dewatering. Within mixed and homogeneous groups, average feeding levels decreased during dewatering. The highest ranking fish, which was always a brown trout in mixed groups, predominantly monopolized the pool and other individuals in pools adopted a more cryptic, stationary behaviour. Dewatering effectively increased local population density with the result that dominance status became much more important in maintaining food intake, and polarization between the top ranking fish and others increased. During the first day of dewatering, there was extreme behavioural polarization such that the dominant fish exhibited most aggression and least feeding within the group. Among dominant fish on the second day of dewatering, aggression had largely abated and feeding had returned to pretreatment levels despite the reduced average feeding within the group. The main difference between mixed and homogeneous groups was in the behaviour of the most dominant Atlantic salmon, which was near-despotic in allopatry and subordinate to brown trout in sympatry.     

Low stocking incidence in brown trout populations from northwestern Spain monitored by LDH-5* diagnostic marker

Brown trout Salmo trutta is one of the most valuable species inhabiting river drainages in Galicia (northwestern Spain). The influence of man, through overfishing and pollution, is thought to have caused the decline of trout in Galician rivers. To balance the possible population decline, Galician rivers have been stocked extensively with a brown trout stock of German origin during the last 30 years. In this study, the incidence of stocking practices has been investigated by an LDH-5 * genetic marker. A very low number of stocked individuals have been observed within river populations, despite the long period of repopulation. Only eight out of the 44 populations analysed showed a limited number of individuals of hatchery origin. Most stocked individuals detected were 0+ age, and showed a poor condition factor ( K < 1). Environmental factors (muscular stamina and food habits) and genetic factors (different selective conditions in hatchery medium) are invoked to explain the low viability of hatchery fish Observed in this Study.     

Multiple electrofishing as a mitigate tool for removing nonnative Atlantic brown trout (<Emphasis Type="Italic">Salmo trutta</Emphasis> L.) threatening a native Mediterranean brown trout population

In the native range of the brown trout (Salmo trutta L.) in Europe, the hybridization of native populations by nonnative domesticated strains introduced by stocking is one of the most serious threats to the long-term conservation of diversity within this species. With the objective of conserving and restoring the native gene pool, fishery managers are beginning to implement various management strategies at the local scale. Nevertheless, few case studies have been published that investigate the effectiveness of the various different conservation strategies for native brown trout populations. In the Chevenne Creek, a small French mountain stream, we tested the strategy of removing nonnative individuals by multiple electrofishing carried out by fishery managers in order to evaluate its feasibility and effectiveness for eliminating a nonnative population threatening a native population. Electrofishing produced major reductions in the nonnative population between 2006 and 2009, with 82–100% of nonnative individuals being removed over a period of 4 years. Nevertheless, despite multiple-electrofishing campaigns, this nonnative population was not entirely eradicated, and some natural recruitment persisted. The young of the year and subadults were less effectively removed than the adults. The results suggest that repeated electrofishing campaigns can be used by managers to reduce the nonnative brown trout population with the objective of conserving the native gene pool, but the removal operation must be continued for at least 4 consecutive years. This strategy, which is feasible in small streams, has to be followed by complementary operations to allow the restoration of a new, native, self-sustainable brown trout population.