Current status of the brown trout (Salmo trutta) populations within eastern Pyrenees genetic refuges

Since the end of the 20th century, some headwaters of rivers in the eastern Pyrenees have been designated as genetic refuges to protect remaining native brown trout (Salmo trutta) diversity. The declaration was based on limited or no evidence of genetic impact from released non‐native Atlantic hatchery fish. Hatchery releases were completely banned into the genetic refuges, but pre‐existing fishing activities were maintained. Specific locations in each refuge have been monitored every 2–3 trout generations to update genetic information to accurately assess the contribution of these reservoirs to the preservation of native brown trout gene pools. This work updates genetic information to year 2014 in three of these locations (in Ter, Freser and Flamisell rivers). Previous studies identified hatchery introgressed populations within refuges and suggested discrepancies between the underlying intention of the genetic refuges and the gene pools detected. Therefore, we also examined genetic divergences among locations inside refuge river segments. Combined information at five microsatellite and the lactate dehydrogenase C (LDH‐C*) loci showed reduced but significant temporal native allele frequency fluctuations in some of the above specific locations that did not modify overall levels of local diversity and river divergences. Bayesian clustering analyses confirmed the presence of differentiated native units within each genetic refuge. Some locations of the Freser River within the genetic refuge area showed high hatchery impact of non‐native fish (over 20%). We discuss additional local actions (releases of native fish, selective removals and fishery reinforcement with sterile individuals) to improve the conservation objective of genetic refuges.     

Competition between hatchery‐raised and wild brown trout Salmo trutta in enclosures – do hatchery releases have negative effects on wild populations?

Abstract –  We compared the growth and prey consumption of juvenile hatchery and wild brown trout of similar genetic origin in 12 wire mesh cages in a natural river in North-Eastern Finland. Wild trout started feeding shortly after the start of the experiment, and clearly earlier than novel hatchery trout, but ate less in the presence of hatchery trout. When accompanied with wild trout, novel hatchery trout started to feed earlier, consumed more live prey, and lost less weight than when in allopatry. Hatchery trout grew more slowly than wild trout. To our knowledge, this is the first study to show that hatchery trout benefit from the presence of wild, experienced trout in a complex semi-natural environment. However, our results also indicate that the benefits to hatchery fish are not transferred to wild fish, and ultimately that care should be exercised in management actions when using hatchery trout to supplement wild populations. On the other hand survival potential of the novel hatchery brown trout could be better in the wild when also undomesticated trout are present.     

Genetic refuges for a self‐sustained fishery: experience in wild brown trout populations in the eastern Pyrenees

Abstract – Management policies balancing harvest and conservation of natural populations of fish are difficult to establish, both scientifically and politically. This issue is particularly difficult when those populations represent native genetic resources. Since 1997, several brown trout populations in the eastern Pyrenees Mountains (Spain) were designated as ‘genetic refuges’ under varying fishing regulations, where releases of hatchery‐origin fish are not permitted. We analysed genetic variation in samples of brown trout from six of those refuge populations and four non‐refuge populations within the same region. Each population was sampled in four separate years: 1993, 1999, 2004 and 2006. Our analyses were based on a diagnostic allele (LDH‐C*90) that distinguishes native and exogenous hatchery populations. Comparisons were based on stocking histories before and after refuge designations and on three management strategies: fished, unfished and catch‐and‐release. Overall, we detected significant genetic introgression resulting from past stocking practices despite the current restriction of hatchery releases imposed by the recent genetic refuge policy. However, this new policy has prevented detectable introgression from increasing throughout the region and together with additional measures on length and number of captured fish is contributing to self‐sustained fisheries that are achieving conservation goals. Quick acceptance of ‘genetic refuges’ by anglers in one particular river, the Ter River basin, has been a key factor in protecting native gene pools compared with the Segre River basins where refuges were not readily accepted.     

Effects of stocking on the genetic structure of brown trout,Salmo trutta,in Central Europe inferred from mitochondrial and nuclear DNA markers

Abstract Stocking has had a considerable effect on wild brown trout, Salmo trutta L., populations throughout Europe. To elucidate this impact and to outline further management strategies, the genetic structure of 25 wild populations and five hatchery stocks from Czech Republic and Slovakia were analysed using mitochondrial (control region) and nuclear DNA (microsatellites, LDH‐C1*) markers. Stocking practices have caused massive hybridisation between the Atlantic and Danube brown trout strains in the central Danube basin and have lead to a loss of among‐population divergence in Slovakia and the eastern part of Czech Republic. Comparison with studies from neighbouring countries revealed substantial differences in haplotype, allele frequencies and genetic diversity across Central Europe. Differences in stocking management and origin of breeding stocks appear to be crucial factors for the spatial variability of the genetic structure of brown trout.     

An electrophoretically-detectable genetic tag for hatchery-reared brown trout (Salmo trutta L.)

The feasibility of incorporating a unique genetic marker into a hatchery strain of brown trout is investigated. The allele Pgi-3(110) is shown to have a very limited distribution among native trout populations in Great Britain and Ireland yet is present, at low frequency, in all three hatchery stocks examined. The potential therefore exists to breed a strain of hatchery brown trout homozygous for the Pgi-3(110) allele. Individuals of such a strain could be unambiguously distinguished from virtually all native stocks. The usefulness of the genetic tag is enhanced by the strong expression of Pgi-3 in adipose fin, permitting simple biopsying. Data from population surveys and the monitoring of experimental progeny suggest selective equivalence among Pgi-3 genotypes.     

Predation on wild and hatchery salmon by non‐native brown trout (Salmo trutta) in the Trinity River,California

Non‐native predators may interfere with conservation efforts for native species. For example, fisheries managers have recently become concerned that non‐native brown trout may impede efforts to restore native salmon and trout in California's Trinity River. However, the extent of brown trout predation on these species is unknown. We quantified brown trout predation on wild and hatchery‐produced salmon and trout in the Trinity River in 2015. We first estimated the total biomass of prey consumed annually by brown trout using a bioenergetics model and measurements of brown trout growth and abundance over a 64‐km study reach. Then, we used stable isotope analysis and gastric lavage to allocate total consumption to specific prey taxa. Although hatchery‐produced fish are primarily released in the spring, hatchery fish accounted for most of the annual consumption by large, piscivorous brown trout (>40 cm long). In all, the 1579 (95% CI 1,279–1,878) brown trout >20 cm long in the study reach ate 5,930 kg (95% CI 3,800–8,805 kg) of hatchery fish in 2015. Brown trout predation on hatchery fish was ca. 7% of the total biomass released from the hatchery. Brown trout only ate 924 kg (95% CI 60–3,526 kg) of wild fish in 2015, but this was potentially a large proportion of wild salmon production because wild fish were relatively small. As large brown trout rely heavily on hatchery‐produced fish, modifying hatchery practices to minimise predation may enhance survival of hatchery fish and potentially reduce the abundance of predatory brown trout.     

Stocking impact,population structure and conservation of wild brown trout populations in inner Galicia (NW Spain), an unstable hydrologic region

1. Brown trout (Salmo trutta) is an important conservation resource in the Iberian Peninsula. The Atlantic is considered the most hydrologically stable region for the species, although inner Galicia (NW Spain) shows Mediterranean (unstable) climatic conditions. The Galician region, threatened by past releases of brown trout individuals from central European origin, harbours two native lineages, one of them endemic to the Iberian Peninsula. These populations are thus highly valuable for conservation, as well as being important for recreational fisheries. 2. In total, 546 individuals from 16 sampling sites (15 natural locations from inner Galicia and one from a central European hatchery stock) were genotyped for 11 nuclear markers (10 microsatellite loci and the LDH‐C* locus) to analyse genetic variability, population structure and introgression impact from stocking in order to assess the conservation status of brown trout in the region. Moreover, correlation among hatchery introgression and environmental variables relevant for species population dynamics was also investigated. 3. Genetic variability was within the range of Iberian brown trout (He = 0.500–0.600). Stocking impact was higher than previously reported values for the Atlantic region and was related to environmental instability. Highly significant native population differentiation was observed in the whole region (F_ST = 0.283), at least four main genetic groups being detected across the geographic distribution studied. 4. Conservation strategies at local level (including the creation of genetic refuges and temporal monitoring of genetic composition) are suggested to agencies and administrations for the sustainable management of brown trout.     

Failure of predator conditioning: an experimental study of predator avoidance in brown trout (Salmo trutta)

Many studies have documented that hatchery‐reared salmonids generally have inferior survival after being stocked compared with wild conspecifics, hatchery and wild salmonids have been observed to differ in their antipredator responses. The response of brown trout (Salmo trutta) juveniles (0+) of differing backgrounds to a live predator was compared in two experiments. First, the antipredator behaviour of predator‐naïve hatchery‐reared brown trout and wild‐exposed brown trout were assessed in behavioural trials which lasted for eight days. Second, predator‐naïve and predator‐conditioned hatchery‐reared brown trout were assessed in identical behavioural trials. Brown trout were ‘predator‐conditioned’ by being held in a stream‐water aquarium with adult Atlantic salmon (Salmo salar) and adult brown trout for two days prior to behavioural trials. Predator‐conditioned hatchery‐reared brown trout spent more time in shelters in the trial aquaria than predator‐naïve hatchery‐reared fish, but did not differ in time spent in the predator‐free area. Predator conditioning may account for the increased time spent in the shelter, but does not appear to have affected time spent in the predator‐free area. However, even if significant alteration in behaviour can be noted in the laboratory, the response might not be appropriate in the wild.     

Genetic characterization of naturalized populations of brown trout Salmo trutta L. in southern Chile using allozyme and microsatellite markers

This study describes the genetic structure of five naturalized populations of brown trout in southern Chile using allozyme and microsatellite markers to establish levels of intra‐ and interpopulation genetic variability and divergence. Fourteen enzymatic systems were used comprising 20 loci and three microsatellite loci specific to brown trout. The genetic variability values (allozymes, P=20–35%, average=27%, H_O=0.118–0.160, average=0.141; microsatellites, P=33.3–100%, average=66.66%, H_O=0.202–0.274, average=0.229) are similar to values described in other naturalized populations of brown trout present in Chile, but higher than those observed in European populations of this species. Values of total genetic diversity (H_T) (allozymes=0.1216 and microsatellites=0.3504) and relative genetic divergence (G_ST) (allozymes=9.5% and microsatellites=15%) were also similar to the results obtained in previous studies of Chilean populations of brown trout. These values, when compared with those obtained in Europe, proved to be similar for H_T but lower for G_ST. The low interpopulational genetic differentiation was in accordance with the small genetic distance observed between the populations analysed (D Nei=0.004–0.025). On the other hand, the high frequency of one of the two alternative alleles of the phylogeographic marker locus LDH‐5* in the populations analysed (LDH‐5*90>0.84) would indicate a European origin, in particular Atlantic as opposed to Mediterranean, for the brown trout introduced into Chile. The high levels of genetic variability suggest a mixed origin for the naturalized brown trout in Chile, which could have originated either before or during the introduction process. Nevertheless, the low level of genetic differentiation between populations could reflect the short lapse of time in evolutionary terms, during which populations introduced into Chile have been exposed to different evolutionary forces, and which has not been sufficiently long to produce greater genetic differentiation between populations.     

Allozyme variation of hatchery and river populations of rohu (Labeo rohita, Hamilton) in Bangladesh

The genetic variations of rohu (Labeo rohita, Hamilton) sampled from five hatchery populations (Arabpur, Brahmaputra, Comilla, Kishorganj and Natore) and three major river populations (the Halda, the Jamuna and the Padma) were analysed by allozyme electrophoresis. Ten enzymes encoded by 11 loci were screened, and six were polymorphic. Alleles at three loci (Est‐1^*, Gpi‐1^* and Gpi‐2^*) proved variable for hatchery and river populations, and the Mdh‐2^* locus exhibited heterozygous genotypes for river populations only. Polymorphic loci per population (27.3±5.3%), heterozygous loci per individual (15.5±1.2%) and relative gene diversity (0.27±0.08) in river populations were higher than those for hatchery populations (25.5±1.8%, 10.7±1.6% and 0.25±0.01 respectively). Also, the observed heterozygosity (Ho) and expected heterozygosity (He) (0.09±0.03 and 0.14±0.04 respectively) in river populations were higher than those in hatchery populations (0.08±0.01 and 0.11±0.01 respectively). The lower levels of genetic variability in hatchery populations suggested the occurrence of inbreeding and/or genetic drift. The pairwise population differentiation (F_ST) values showed a lower level of genetic differentiation between hatchery and river population pairs. The unweighted pair‐group method with arithmetic mean dendrogram of Nei's genetic distances showed a relationship between the genetic distance and geographic distance. The populations were clustered into three groups: the Padma in one group, the Halda in second group and the Jamuna, including five hatcheries, in the third group. Highly diversified rohu individuals were observed in the Padma and Halda Rivers, whereas less genetically variable individuals were found in the Jamuna River and five hatcheries. These findings can be useful for rohu hatchery propagation to enhance the sustainable aquaculture production.     

Conservation of endemic landlocked salmonids in regulated rivers: a case‐study from Lake Vänern,Sweden

Conservation of migratory salmonids requires understanding their ecology at multiple scales, combined with assessing anthropogenic impacts. We present a case‐study from over 100 years of data for the endemic landlocked Atlantic salmon (Salmo salar, Salmonidae) and brown trout (Salmo trutta, Salmonidae) in Lake Vänern, Sweden. We use this case‐study to develop life history‐based research and monitoring priorities for migratory salmonids. In Vänern, small wild populations of salmon and trout remain only in the heavily regulated Rivers Klar (Klarälven) and Gullspång (Gullspångsälven), and commercial and sport fisheries are maintained by hatchery stocking. These populations represent some of the last remaining large‐bodied (up to 20 kg) landlocked salmon stocks worldwide. We found that one of four stocks of wild fish has increased since 1996; the other three remain critically low. Hatchery return rates for three of four stocks appear stable at roughly 1% and annual fisheries catch is roughly 75 metric tons, with an estimated 7.5% of hatchery smolts being recruited to the fishery; this also appears relatively stable since 1990. Our analysis reveals much uncertainty in key data requirements, including both river return and fisheries catch rates, estimates of wild smolt production and survival, and hatchery breeding and genetics protocols. These uncertainties, coupled with a lack of information on their riverine and lacustrine ecology, preclude effective management of these unique populations. We conclude with a framework for a life history‐based approach to research and monitoring for Vänern salmon and trout, which should be applicable for all endemic, migratory salmonid populations.     

Genetic characterization of Portuguese brown trout (Salmo trutta L.) and comparison with other European populations

Abstract– Allozyme and other protein loci were examined to study the genetic structure of Portuguese brown trout ( Salmo trutta ) populations. A total of 247 individuals from three tributaries of the Lima hydrological basin and a hatchery, all located in northern Portugal, were analyzed. Four of 22 protein coding loci were found to be polymorphic: CK-A1^*, GPI-A2^*, MPI-2^* and TF^*. A new allelc at the latter locus was found in Atlantic populations. The data obtained for Portuguese brown trout were compared with published data for 14 European populations and three hatchery stocks. Six polymorphic loci (CK-A1^*, GPI-A2^*, GPI-B2^*, LDH-C^*, ME^* and MPI-2^*) were used in a cluster analysis. This showed the similarity of Portuguese natural populations and northern Iberian populations and that Portuguese hatchery fish have an autochthonous origin, distinct from that of other Atlantic hatchery stocks.     

Genetic Introgression Between Arctic Charr (Salvelinus Alpinus) and Brook Trout (Salvelinus Fontinalis) in Bavarian Hatchery Stocks Inferred from Nuclear and Mitochondrial DNA Markers

Nuclear insulin-like growth factor 2 gene (IGF-2), growth hormone 1 gene (GH-1) and internal transcribed spacer 1 (ITS-1) of the ribosomal DNA as well as the mitochondrial NADH-3 and NADH-4 dehydrogenase genes (ND-3/4) exhibited species-specific restriction fragment patterns and three microsatellite loci (Sfo18, Ssa85 and Ssa197) had non-overlapping allele size ranges in Arctic charr and brook trout and were used as diagnostic markers for testing genetic purity of hatchery stocks and wild populations of Arctic charr and brook trout in Bavaria, Germany. Screening of four wild populations (three in Arctic charr and one in brook trout) revealed only a single hybrid (back-cross to brook trout) individual in L. Starnberg. In contrast, in three (out of five) hatchery stocks of Arctic charr and in both hatchery stocks of brook trout hybrids were detected with the frequency from 3 to 100%. Three hatchery stocks (SS2, SA and BS1) represent a hybrid swarm because they contained a very high proportion of hybrids (from 83 to 100%) and most or all hybrid individuals had alien alleles at only one or a few of six unlinked diagnostic loci, indicating that post-F₁ hybrids represent the majority of individuals in these stocks and introgression has taken place. Release or escape of introgressed individuals from hatcheries into natural water bodies should be avoided in order to protect the biological diversity and genetic integrity of native fish populations.     

Feeding by hatchery-reared and wild brown trout, Salmo trutta L., in a Norwegian stream

Abstract. Hatchery-reared brown trout, Salmo trutta L., yearlings were captured shortly (3h to one week) after their release in a Norwegian stream. The feeding of recaptured hatchery fish was compared with that of wild brown trout. The investigations were carried out during three different periods (May, July and October). Investigations of drift fauna indicated that food availability was best in May. Most hatchery-reared brown trout started feeding shortly after their release in all three periods. Hatchery fish went through a learning process with respect to feeding. This was most clearly demonstrated by the amounts of plant fragments in their stomachs, which were always greater in hatchery fish than in wild fish but which decreased with time after release in hatchery fish stomachs in all three periods. By about a week after release, hatchery trout appeared to be feeding on wild prey nearly as well as did wild fish, but they achieved this better in May than in October.     

Importance of life‐history and landscape characteristics for genetic structure and genetic diversity of brown trout (Salmo trutta L.)

Abstract – Investigating the influence of evolutionary forces on the genetic structure and genetic diversity remains a major challenge. Yet, it is of considerable interest for conservation and management of a species. This study investigates the influence of life‐history and landscape features, such as altitude, connectivity and habitat size, on genetic diversity and genetic structure of brown trout (Salmo trutta L.) with stream‐resident, lake‐dwelling and sea‐migrating life‐history in two river systems in northern Sweden. Using regression tree analysis including ecological and landscape characteristics, we show that life history is the most important variable explaining genetic diversity and population differentiation. Sea‐migrating populations show high diversity and low differentiation, and lake‐ and stream‐resident populations show low diversity and high population differentiation, among all samples. No overall genetic correlation with geographical distance was noted; however, among sea‐migrating populations within the River Vindelälven drainage, this pattern was observed. This study illustrates that life‐history and landscape features help to explain genetic structure and genetic variation. The information is important for conservation and management actions, such as fisheries regulations, habitat restorations, stocking of hatchery fish, defining management units and introducing genetic monitoring programmes.     

Genetic effects of introduced hatchery stocks on indigenous brown trout (Salmo trutta L.) populations in Spain

Abstract– To assess the levels of gene introgression from cultured to wild brown trout populations, four officially stocked locations and four nonstocked locations were sampled for one to three consecutive years and compared to the hatchery strain used for stocking. Allozyme analysis for 25 loci included those previously described as providing allelic markers distinguishing hatchery stocks and native populations. Different levels of hybridization and introgression with hatchery índividuals were detected in stocked drainages as well as in protected locations. These findings indicate that new policies for stocking and monitoring hatchery fish are needed if gene pools of wild Spanish brown trout populations are to be preserved.     

Allozyme diversity in Australian rainbow trout, Oncorhynchus mykiss

Rainbow trout, Oncorhynchus mykiss (Walbaum), were first introduced into Australia over 100 years ago, and forms the basis of important recreational inland fisheries and an aquaculture industry in south‐eastern Australia. This paper investigates the genetic variation within and between samples of Australian rainbow trout using allozyme electrophoresis. The levels of genetic diversity within Australia do not show marked differences from those observed in hatchery and wild populations from throughout North America, New Zealand and South Africa, but there is evidence for the loss of some rare alleles during translocation from California to Australia via New Zealand. No appreciable difference in genetic diversity was apparent between hatchery and self‐sustaining wild populations of rainbow trout from mainland Australia. However, significant differences in allelic frequencies were observed, with consistent genetic differences between Victorian and New South Wales samples most likely reflecting state‐based hatchery and stocking policies.     

Erosion of the native genetic resources of brown trout in Spain

Abstract– We analyzed the introduction of hatchery-reared trout in the Riutort Creek, a small stream in the eastern Spanish Pyrennees. We used gene correlation matrices between individuals to analyze the fish coancestry in the Riutort Creek samples and in the hatchery stock. Hatchery fish disturbed the single ancestry in the native population of the creek, and were clearly detected with principal coordinate analysis of the gene correlation matrix. The amount of introgression produced by successful introductions was estimated from the principal coordinate analysis projections of the matrix of F_ST values between the putative native Riutort Creek population, the hatchery stock and the introgressed population. In only two years the amount of introgression rose to 10%, indicating that 5% of the native ancestry is lost each year as a result of the stocking program. Based on these results, we review the present understandings on the genetic impact of hatchery fish on indigenous Spanish brown trout populations. The stocking of these populations involves a non-native broodstock widespread through the Spanish hatcheries, but successful stockings appear to be limited to wild populations subjected to occasional releases in protected or unfished areas. Surprisingly, extensive stocking in fished areas result in a more limited genetic impact on the recipient native population.     

Disentangling individual movement between populations from effective dispersal in the facultative anadromous Salmo trutta L.

Gathering information on both individual movement and gene flow is rarely possible when studying dispersal among populations in fish species. It is, however, possible to assess both at a reasonable cost in Salmo trutta L. on the Atlantic coast of Europe where the facultative anadromous species is composed of discrete populations of brown trout residents occupying distinct river systems, but exchanging phenotypically distinguishable sea trout migrants. We performed two kinds of genetic analyses using individual microsatellite genotypes: the stock identification of sea trout entering each corridor and the estimates of effective dispersal through each corridor. We observed that individual movement (nonlocal individuals of each source population ranging from 4% to 35% of the sea trout run) never translates into effective dispersal except in one of four migratory corridors examined. The likely origin of this uniquely detected gene flow event is discussed in the light of well‐documented migratory fish management actions undertaken in the past in the studied area.     

Do dams increase genetic diversity in brown trout (Salmo trutta)? Microgeographic differentiation in a fragmented river

Abstract – Local genetic differentiation may potentially arise in recently fragmented populations. Brown trout is a polytypic species exhibiting substantial genetic differentiation, which may evolve in few generations. Movement (semi‐)barriers in rivers may cause fragmentation, isolation and genetic differentiation in fish. In the Måna River (28 km) flowing from the alpine Lake Møsvatn to the boreal Lake Tinnsjø, construction of four hydropower dams during the period 1906–1957 have fragmented the previously (since last Ice Age) continuous wild resident brown trout population. Samples from the two lakes (N = 40) and six sites in the river (N = 30) isolated at different times were analysed at nine microsatellite loci. All populations showed substantial genetic variation (mean number of alleles per locus 5.3–8.9, observed heterozygosity 0.57–0.65 per population, overall F_st = 0.032). Pairwise multilocus F_st estimates indicated no significant differentiation between populations in the two lakes, and no or little differentiation in the lower river (F_st = 0.0035–0.0091). The microgeographic differentiation among wild resident trout at these sites was less than expected based on similar previous studies. However, results from the upper river, in particular the site immediately below the Lake Møsvatn outlet and dam, indicated isolation (F_st > 0.035). Calculation of genetic distances and assignment tests corroborated these results, as did a significant correlation between years of isolation (since dam construction) and F_st. The population structuring is most likely a result of fragmentation by dams, which has increased overall genetic diversity. This increased local differentiation may be caused by natural selection, but more likely by genetic drift in small, recently fragmented populations. Increased local genetic diversity by genetic drift does not justify conservation measures aiming at preserving genetic diversity.