Genetic structure and demographic history of brown trout (Salmo trutta) populations from the southern Balkans

1. The present study was designed to characterize the genetic structure of brown trout ( Salmo trutta ) populations from the southern Balkans and to assess the spread of non-native strains and their introgression into native trout gene pools. We analysed polymorphism at nine microsatellite loci in seven supposedly non-admixed and three stocked brown trout populations.
2. The analyses confirmed the absence of immigration and extraordinarily strong genetic differentiation among the seven non-introgressed populations in parallel with low levels of intrapopulation genetic variability. In contrast, analyses of the stocked populations revealed that the genetic integrity of the local populations had been substantially changed, and the populations must be characterized as hybrid swarms. The pattern of population differentiation observed at microsatellites contrasted to that depicted previously by mtDNA variation. However, the close relationships between populations from the Danube and Axios river systems proposed solely by microsatellites could be explained by palaeogeographic events.
3. Our research showed that most of the populations examined represent unique gene pools, whose existence is critically compromised. Therefore, appropriate management and conservation strategies should be developed urgently in order to protect the subspecific biodiversity and to reverse currently negative trends.     

Genetic detection of sex-specific dispersal in historical and contemporary populations of anadromous brown trout Salmo trutta

The study of sex-biased dispersal has attracted considerable attention in birds and mammals, but less in other taxa, including fishes. We analysed sex-specific dispersal in historical (1910s and 1950s) and contemporary (1990s) samples of anadromous brown trout Salmo trutta. We tested the hypothesis that dispersal is unbiased using information from microsatellite DNA and applying an assignment index for 11 temporally and spatially separated samples. Our results are most consistent with brown trout dispersal being male biased, and provide no evidence of female bias. We found no evidence that dispersal patterns changed over time, indicating that stocking with hatchery strains did not affect sex-specific dispersal.     

Genetic divergence among native trout Salmo trutta populations from southern Balkans based on mitochondrial DNA and microsatellite variation

The genetic structure and the phylogenetic relationships among five Balkan populations of trout Salmo trutta that have been classified earlier into five different taxa were studied, using microsatellite and mitochondrial DNA (mtDNA) analyses. The pattern of population differentiation observed at microsatellites differed to that depicted by mtDNA variation, yet both methods indicated a very strong partitioning of the genetic variation among sampling locations. Results thus suggest that conservation strategies should be directed towards preserving the genetic integrity and uniqueness of each population.     

Allozyme diversity in brown trout (Salmo trutta) from Central Spain: Genetic consequences of restocking

1.  The brown trout ( Salmo trutta ) represents one of the main freshwater resources in Spain, but habitat alterations and overharvesting have contributed to the decline or disappearance of numerous natural populations. In addition, reinforcement programs of wild populations based on releases of hatchery reared fish of exogenous origin compromise the conservation of remnant native trout resources.
2.  We present allozymic data from Central Spain trout populations including stocked and unstocked populations. Although the levels of genetic variation observed were low and affected by hatchery releases (p = 18.23%, Ho= 3.39%), they were within the range observed in other European areas.
3.  The effective introduction of hatchery reared fish is genetically homogenising the populations in the studied area and disturbing the ancestral pattern of genetic variation that distinguishes the Tajo and Duero basins. Within the eight natural populations analysed, seven had alleles assigned to the foreign trout. The introgression in these populations, following the LDH-5 * 90 allele frequency, ranged between 2% and 29.4%, but those values are not in concordance with the respective stocking effort undertaken in each population. Moreover, the release of hatchery-reared fish does not solve the problems related to the reduced size of wild populations and their recruitment instability.     

Electrophoretic variation in six populations of brown trout (Salmo trutta L.)

Starch gel electrophoretic studies of 16 enzymes encoded by 34 Loci were performed on six brown trout populations. One new polymorphism is described at the Pmi-2 locus. Breeding data were analysed for both single and joint segregation of six loci: Aat-1, Cpk-1, G3p-2, Mdh-2, Mdh-3, and Pmi-2. All the loci are shown to segregate in simple mendelian ratios and one nonrandom joint segregation was observed. The polymorphism level, heterozygosities, and genetic distances were estimated and compared with those reported in other studies on brown trout and closely related salmonid species. The polymorphism level (25%) and average heterozygosity (9%) were high. Significant genetic distances were observed, but the average degree of differentiation between populations appeared to be small (9% of the total heterozygosity).     

Structure and genetic diversity of small populations of brown trout Salmo trutta in Kandalaksha Bay,the White Sea

The phenetic variability (types of life strategy, age and sex structure of population, biological peculiarities), the anthropogenic pressure, and the genetic diversity were studied by five microsatellite loci for several minor populations of brown trout Salmo trutta inhabiting the waterbodies of Velikaya Salma Strait watershed area (Kandalaksha Bay, the White Sea). The level of genetic and phenetic diversity in small populations was comparable to those representing large water systems. Significant spatial dissimilarity of the populations located in less than 15-km distance was found. Low allelic and genetic diversity was observed for the studied populations. Variability of some studied loci through the time period was observed for the fish sampled from different populations. We assume that minor populations of brown trout may be treated as the standards of native populations for the monitoring activities on the brown trout populations and its environment both in short-term and long-term perspective.     

Genetic introgression between native and introduced brown trout Salmo trutta L. populations in the Rhone River Basin

In the Doubs River (Rhône drainage) two distinct brown trout ( S. trutta ) phenotypes are observed. One phenotype is locally called Doubs trout and is characterized by four black stripes on the sides, similar to perch ( Perca fluviatilis L.) and the other is the common phenotype of the fluviatile ecotype of brown trout, Salmo trutta f. fario . Protein data for three samples from the Doubs show that the Doubs trout belongs to the Mediterranean population group of brown trout, whereas the fario phenotype originates from stocking with hatchery strains of Atlantic basin origin. The two forms, however, do not hybridize freely. This is indicated by considerable gametic phase disequilibrium between alleles of hatchery and Doubs trout at one sampling site, and by lack of intermediate genotypes and phenotypes at another sampling site. The introgression patterns observed at the two sites suggest that differences in local habitat conditions can affect the degree of hybridization and introgression.     

Thermoregulatory behavior of brown trout,Salmo trutta

Brown trout, Salmo trutta, were allowed to thermoregulate individually in an electronic shuttlebox. Pooled data for 6 fish showed a diel pattern of preferred temperature, with a diurnal minimum of 10.3°C, an early nocturnal maximum of 13.7°C, a less pronounced mid-scotophase minimum of 11.7°C, and a secondary dawn maximum of 12.8°C, in a somewhat crepuscular pattern. The 24-hour mean preferendum was 12.2°C.     

Genetic differentiation among the sympatric brown trout (Salmo trutta) populations of Lough Melvin, Ireland

Three morphotypes of brown trout have been described from Lough Melvin in north-west Ireland: gillaroo; sonaghen; ferox. The extensive genetic differences among the three types indicate that they are reproductively isolated, i.e. separate stocks of trout. For example, only gillaroo possess the Ldh -1(n) allele. Ferox show a high frequency of the Ldh -5(100) allele and this allele does not exceed a frequency of 0.02 in the other two types. Sonaghen are characterized by relatively much higher frequencies for Ck -2(115) and Gpi- 2(135) alleles. Sampling over a period of 7 years and experiments with artificial stocks of gillaroo and sonaghen have demonstrated that these differences are temporally stable. On the basis of observation of mature adults and the genetic composition of fry samples, it has been shown that the three types maintain their genetic integrity as the result of their distinctive spawning habits with gillaroo spawning in the lake and outflowing river, sonaghen in the smaller inflowing rivers and ferox in the deep downstream section of the largest inflowing river. The ferox is a relict population of an early post-glacial trout colonist while gillaroo and sonaghen are representative of a more recent independent colonization. However, it cannot be determined at present whether or not gillaroo and sonaghen were already differentiated prior to colonizing Melvin although the weight of evidence favours a sympatric split. It is proposed that the three types of trout be designated as separate subspecies to highlight the need for independent management and urgent requirement for conservation action.     

Genetic structure in alpine brown trout Salmo trutta L. shows that indirect stocking affects native lake populations

Genetic variation of brown trout inhabiting six connected lakes (Krokavatn, Skavatn, Kollsvatn, Litlosvatn, Kvennsjøen and Øvre Krokavatn) of the Hardangervidda mountain plateau was analysed by 11 microsatellites covering a period of 24–35 years. Temporal changes were found and are deemed to be mostly related to stocking in the headwater lakes Krokavatn and Skavatn. A pronounced change was evident in one population (Kollsvatn) and was the result of downstream migration of stocked fish from these headwater lakes. This indirect stocking accounted for up to 50% in the STRUCTURE analysis of Kollsvatn fish and was less pronounced in the other downstream populations of Litlosvatn ( c. 28%), Kvennsjøen ( c. 17%) and Øvre Krokavatn ( c. 13%). Since significant downstream migration of stocked fish affecting native populations occurs, nature management efforts must respond accordingly.     

Allozyme variability in brown trout Salmo trutta in Chile

1. Brown trout ova were imported during the last century from different locations in Europe to establish populations in Chilean rivers (South America). The rivers are currently occupied by naturalized populations that have adapted to very different environmental conditions, such as areas of semi-desert in the north, or rainy and cold areas in the south.
2. In this first study in this geographical area, electrophoretic variability of proteins encoded by twenty-five loci was screened in seven populations from northern to southern Chile.
3. The results show significant heterogeneity of allelic frequencies between populations in seven of eleven polymorphic loci detected. The estimated value of genetic diversity 0.1274 ( H _T) is higher than that observed in populations from areas of natural distribution of this species. However, only 12.64% of this genetic diversity was found between samples ( G _ST), indicating a low genetic divergence among Chilean populations. The observed associations among the Chilean and 'modern' group of European populations suggests the probable origin of the new populations.     

Sexual size dimorphism in anadromous brown trout Salmo trutta

Anadromous trout Salmo trutta exhibits sexual size dimorphism (S_SD); females were larger than males in populations where male mean total length (L_T) at maturity was below 49 cm and females were smaller than males when mean male L_T was above 49 cm, the slope of the regression of female on male L_T was 0·59. In streams with mean annual discharge below 41 m³ s^?1, flow added significantly to a model with S_SD as the dependent variable and male mean L_T at maturity as the first predictor variable. There was a slight increase in S_SD with increasing latitude, which may result from an increase in male size with increasing latitude.     

Impacts of acidification on brown trout Salmo trutta populations and the contribution of stocking to population recovery and genetic diversity

Anthropogenic acidification in SW-Scotland, from the early 19th Century onwards, led to the extinction of several loch (lake) brown trout (Salmo trutta) populations and substantial reductions in numbers in many others. Higher altitude populations with no stocking influence, which are isolated above natural and artificial barriers and subjected to the greatest effect of acidification, exhibited the least intrapopulation genetic diversity (34% of the allelic richness of the populations accessible to anadromous S. trutta). These, however, were characterised by the greatest interpopulation divergence (highest pairwise D_EST 0.61 and F_ST 0.53 in contemporary samples) based on 16 microsatellite loci and are among the most differentiated S. trutta populations in NW-Europe. Five lochs above impassable waterfalls, where S. trutta were thought to be extinct, are documented as having been stocked in the late 1980s or 1990s. All five lochs now support self-sustaining S. trutta populations; three as a direct result of restoration stocking and two adjoining lochs largely arising from a small remnant wild population in one, but with some stocking input. The genetically unique Loch Grannoch S. trutta, which has been shown to have a heritable increased tolerance to acid conditions, was successfully used as a donor stock to restore populations in two acidic lochs. Loch Fleet S. trutta, which were re-established from four separate donor sources in the late 1980s, showed differential contribution from these ancestors and a higher genetic diversity than all 17 natural loch populations examined in the area. Genetically distinct inlet and outlet spawning S. trutta populations were found in this loch. Three genetically distinct sympatric populations of S. trutta were identified in Loch Grannoch, most likely representing recruitment from the three main spawning rivers. A distinct genetic signature of Loch Leven S. trutta, the progenitor of many Scottish farm strains, facilitated detection of stocking with these strains. One artificially created loch was shown to have a population genetically very similar to Loch Leven S. trutta. In spite of recorded historical supplemental stocking with Loch Leven derived farm strains, much of the indigenous S. trutta genetic diversity in the area remains intact, aside from the effects of acidification induced bottlenecks. Overall genetic diversity and extant populations have been increased by allochthonous stocking.     

Frequency distribution of coping strategies in four populations of brown trout (Salmo trutta)

In a challenging situation some animals respond by active avoidance, aggression and an activation of the sympathetic nervous system whereas others respond by immobility, low levels of aggression and a predominant adrenocortical stress response. When consistent over time and across situations such inter-individual differences in behavioural and physiological stress responses are referred to as stress coping strategies. In a previous study we reported the existence of two distinct stress coping strategies in a sea-ranched brown trout (Salmo trutta) population. Using the same method, we here show that four brown trout populations with different origin, but reared under identical conditions, differ in their endocrine stress response, behaviour during hypoxia and aggression. Further more, if individuals are classified as high- and low responsive based on post-stress blood plasma noradrenalin levels (indicator of sympathetic reactivity) the frequency distribution shows that populations with hatchery origin are biased towards having higher frequencies of high responsive individuals. However, the number of high responsive trout ranges from 14-48% in the different populations which shows that generally the frequency is biased towards lower levels of high responsive individuals. We discuss different frequency-dependent mechanisms that maintain multiple phenotypes in populations and speculate about differences in selection regime among the studied populations.     

Morphological differences in the skin of marble trout Salmo marmoratus and of brown trout Salmo trutta

Despite being genetically very closely related, the marble trout Salmo marmoratus and the brown trout Salmo trutta exhibit marked phenotypic differences, particularly with regard to skin pigmentation. Histological analysis of skin from the head and gill cover of differently aged individuals of the two species was carried out in order to characterize differences in skin structure. The basic structure of skin of the individuals studied corresponded with that described for other salmonids, though the head epidermis was somewhat thicker in S. marmoratus than in S. trutta, thickening with age in both species. Numerous secretory goblet cells and sporadic secretory sacciform cells were observed in the upper and middle part of the epidermis in both species. Melanophores were present in both species only in the dermis, and were bigger in S. marmoratus and present at lower average density than in S. trutta, and more or less constant across all age classes. In adult S. marmoratus with fully established marble pigmentation, light areas at low density with small (i.e. aggregated) melanophores were present, while in S. trutta melanophores were more uniformly distributed.     

Dynamic micro-geographic and temporal genetic diversity in vertebrates: the case of lake-spawning populations of brown trout (Salmo trutta)

Conservation of species should be based on knowledge of effective population sizes and understanding of how breeding tactics and selection of recruitment habitats lead to genetic structuring. In the stream‐spawning and genetically diverse brown trout, spawning and rearing areas may be restricted source habitats. Spatio–temporal genetic variability patterns were studied in brown trout occupying three lakes characterized by restricted stream habitat but high recruitment levels. This suggested non‐typical lake‐spawning, potentially representing additional spatio–temporal genetic variation in continuous habitats. Three years of sampling documented presence of young‐of‐the‐year cohorts in littoral lake areas with groundwater inflow, confirming lake‐spawning trout in all three lakes. Nine microsatellite markers assayed across 901 young‐of‐the‐year individuals indicated overall substantial genetic differentiation in space and time. Nested gene diversity analyses revealed highly significant (≤P = 0.002) differentiation on all hierarchical levels, represented by regional lakes (F_LT = 0.281), stream vs. lake habitat within regional lakes (F_HL = 0.045), sample site within habitats (F_SH = 0.010), and cohorts within sample sites (F_CS = 0.016). Genetic structuring was, however, different among lakes. It was more pronounced in a natural lake, which exhibited temporally stable structuring both between two lake‐spawning populations and between lake‐ and stream spawners. Hence, it is demonstrated that lake‐spawning brown trout form genetically distinct populations and may significantly contribute to genetic diversity. In another lake, differentiation was substantial between stream‐ and lake‐spawning populations but not within habitat. In the third lake, there was less apparent spatial or temporal genetic structuring. Calculation of effective population sizes suggested small spawning populations in general, both within streams and lakes, and indicates that the presence of lake‐spawning populations tended to reduce genetic drift in the total (meta‐) population of the lake.     

Organic pollution induces domestication-like characteristics in feral populations of brown trout (Salmo trutta)

Sewage pollutants may impair growth or survival of the freshwater biota, though animals might benefit from the extra food availability as production increases. We examined biochemical (muscle glycogen), morphological (condition factor and hepatosomatic index), and diet biomarkers in brown trout for evaluating the effects of chronic exposure to organic pollution. Trout were collected at three locations: ST1 downstream of a trout farm, ST2 affected by the effluents of a wastewater treatment plant and ST3, the reference site. Individuals at polluted sites showed high hepatosomatic index, although no differences were found between ST2 and ST3 for the condition factor. A significant reduction was detected in the levels of muscle glycogen of individuals captured at polluted sites. Moreover, trout diet in these rivers was dominated quantitatively by Chironomidae and Simuliidae, in contrast with the diverse diet of individuals at ST3. Remarkably, individuals at polluted sites showed high stomach fullness and energy gut values, which might be considered as a case of hyperphagia. Our findings suggest that food surplus in organic enriched sites, in the form of high densities of macroinvertebrates, provide an environment similar to that of domesticated animals, where individuals might adopt less energetically costly behavioural strategies to ingest more food.     

Morphological variability among seven populations of brown trout, Salmo trutta L., in Greece

Morphological variation in five meristic and 21 morphometric characters was examined in the brown trout, Salmo trutta L., from seven distinct streams and rivers from N and NW Greece. Variation within each sample, estimated as the multivariate generalization of the coefficient of variation, did not vary substantially among the populations. The phenetic relationships of these populations were examined using the stepwise discriminant analysis. The taxonomic implications of these results are discussed.     

Genetic structure of northwestern Spanish brown trout (Salmo trutta L.) populations, differences between microsatellite and allozyme loci

Genetic variation in nine wild brown trout (Salmo trutta L.) populations was studied by means of allozyme and microsatellite markers. All brown trout populations were clearly separated into two clusters that represented the Sil and Duero basins. Although both markers revealed a strong genetic differentiation between basins, microsatellite loci resulted much more accurate when population structure at the intrabasin level was analysed. Also pairwise multilocus FST estimates and assignment tests of individual fish to the set of sampled populations demonstrated a much higher efficiency of microsatellites compared to allozymes. The analysis of both markers provides new insights in defining the conservation units at this local area and confirms the existence of a recognized sub-lineage in the Duero basin. The management implications of these findings are discussed and changes in trout release activity are recommended to avoid mixing of trout gene pools mainly in the Sil basin.