Secondary contact between two divergent lineages of grayling Thymallus in the lower Enisey basin and its taxonomic implications

A contact zone between two widespread divergent lineages of grayling Thymallus sp. has been identified in the lower Enisey basin (Khantaiskoye Lake) in north central Siberia (Russia). Sequences of the mitochondrial DNA control region revealed two lineages with 2·6% divergence. Haplotypes from these two lineages clearly fall within two previously described clades, one presumably corresponding to Arctic grayling Thymallus arcticus found throughout the Siberian coastal zone as well as North America, and the other corresponding to grayling found elsewhere in the Enisey basin, including Lake Baikal and its tributaries. Allelic variation at seven microsatellite loci strongly suggests reproductive isolation in sympatry between these two lineages in Khantaiskoye Lake. Integration of these new data with existing knowledge on Thymallus throughout its Siberian range supports a taxonomic revision. The currently recognized sub-specific lineages of Arctic grayling along the Siberian Arctic coast as well as grayling from North America should probably be regarded as T. arcticus . Arctic grayling, however, are limited to the coastal zone and lower reaches of the Enisey. Most of the Enisey basin, including Lake Baikal and its tributaries are occupied by another taxon, which should be recognized as Thymallus baicalensis . This perspective strictly follows evolutionary principles, eliminates the use of four sub-specific names and circumvents the illogical mosaic distribution of T. arcticus arcticus , which nevertheless does not represent a monophyletic lineage.     

The effects of fisheries management on harvest rates of native and non-native salmonid fish species

In central Europe, both brown trout Salmo trutta and European grayling Thymallus thymallus are threatened native salmonid species with high value in recreational angling and nature conservation. On the other hand, rainbow trout Oncorhynchus mykiss and brook trout Salvelinus fontinalis are intensively stocked non-native species of high angling value but no value for nature conservation. This study tested if harvest rates of native salmonids are negatively correlated to intensive stocking and harvest rates of non-native salmonids in inland freshwater recreational fisheries. Data were collected from 250 fishing sites (river and stream stretches) over 13 years using mandatory angling logbooks. Logbooks were collected from individual anglers by the Czech Fishing Union in the regions of Prague and Central Bohemia, Czechia (central Europe) and processed by the author of this study. In result, anglers harvested 200,000 salmonids with total weight of 80 tons over 13 years. Intensive stocking of multiple salmonid species lead to slightly lower harvests of native salmonids. Inversely, intensive harvests of multiple salmonid species lead to slightly higher harvest of native salmonids. Recapture rates of stocked salmonids were relatively low (0.6%–3.7%), proving fish stocking moderately ineffective. Since the effects of non-native salmonid stocking and harvest rates on native salmonid harvest were significant but not strong, it is suggested that rivers and streams that support fishing for non-native salmonids still support fishing for native salmonids. However, this idea does not apply for fishing sites with really high intensity of non-native salmonid stocking – harvest rates of natives were very low on these fishing sites.     

Status, habitat use, and vulnerability of the European ggrayling in Austrian waters

The European grayling Thymallus thymallus is widely distributed in Austria, occurring in all eight rural provinces. However, in recent years, an increasing number of studies report severe declines in population sizes. Since 1997, the grayling has had the status of an endangered species in Austria. In 1997 the multidisciplinary research programme 'Local Adaptation, Threat, and Conservation of European Grayling' has carried out three projects in Upper Austria, Salzburg, and Carinthia. Research has included repeated electro-fishing in selected stretches of eight rivers and the collection of data on habitat characteristics, fish species composition, population abundance and size distribution, growth and body condition, as well reproductive timing, and migratory activities of grayling. Furthermore, genetic and morphological variation among grayling populations has been studied as well as the number, habitat use and feeding activity of cormorants. Evidence is provided for the existence of distinct negative effects caused by single factors or combinations of factors on grayling stocks in each of the river stretches studied.     

Multiple species of grayling (Thymallus sp.) found in sympatry in a remote tributary of the Amur River

Large-scale phenotypic and genetic studies of the salmonid genus Thymallus (grayling) in the Palaearctic suggest that most major phylogeographic lineages represent good biological species. Evaluating such a premise in areas involved in palaeo-hydrological dynamics where multiple species are found in sympatry should serve to assess the level of reproductive isolation, the traditional sine qua non of species recognition. Molecular sequence (mtDNA) and microsatellite (nDNA, seven loci) analysis of grayling in the upper Bureya River watershed support the occurrence of three distinctive species of grayling living in sympatry in this large oligotrophic tributary of the Amur River. One of these lineages is primarily found throughout the Lena River basin and is recognized as Baikal-Lena grayling Thymallus baicalolenensis; the second, the upper Amur grayling Thymallus grubii is found over large areas of the Amur catchment including the entire headwater region; and the third, the Bureya grayling Thymallus burejensis is endemic to the study area. A limited number of hybrids were identified, primarily between Baikal-Lena grayling T. baicalolenensis and Bureya grayling T. burejensis with little to no signs of introgression among non-hybrid individuals. Morphological distinctiveness among populations of these species living in sympatry was greater than between populations living in allopatry, suggesting character displacement. Divergence estimates among taxa range up to 6.2 MY, and allopatric origins for all three species’ is suggested. To our knowledge, this is the first data-based confirmation of three species of grayling living in sympatry.     

Genetic Parentage Analysis Confirms a Polygynandrous Breeding System in the European Grayling (Thymallus thymallus)

Knowing the breeding system of a species is important in order to understand individual variation in reproductive success. Large variation in reproductive success and thus reproductive skew strongly impacts on the effective number of breeders and thus the long-term effective population size (N_e). Fishes, in particular species belonging to the salmonid family, exhibit a wide diversity of breeding systems. In general, however, breeding systems are rarely studied in detail in the wild. Here we examine the breeding system of the spring-spawning European grayling Thymallus thymallus from a small Norwegian stream using parentage assignment based on the genotyping of 19 polymorphic microsatellite loci. In total 895 individual grayling fry and 154 mature grayling (57 females and 97 males) were genotyped. A total of 466 offspring were assigned a father, a mother, or a parent pair with a confidence of 90% or higher. Successfully reproducing males had on average 11.9 ± 13.3 (SD) offspring with on average 2.1 ± 1.2 partners, whereas successful females had on average 9.5 ± 12.8 offspring and 2.3 ± 1.5 partners. Parents with more partners also produced more offspring. Thus the grayling breeding system within this small stream revealed a polygynandrous breeding system, similar to what has been observed for many other salmonid fish species. The present study thus unambiguously corroborates a polygynadrous breeding system in the European grayling. This knowledge is critical for managing populations of this species, which has suffered significant local population declines throughout its range over the last several decades.     

New data about distribution of lower Amur grayling <Emphasis Type="Italic">Thymallus tugarinae</Emphasis> and Baikal-Lena grayling <Emphasis Type="Italic">Thymallus</Emphasis> sp. (Thymallidae) in the basins of the upper Amur River and Zeya River

New findings of lower Amur grayling (Lower Amur grayling) Thymallus tugarinae and Baikal-Lena grayling Th. sp. (Th. baicalolenensis) in the basins of the upper Amur River and Zeya River are reported for 2010?2014. Lower Amur grayling has been registered in the tributaries of Argun River and in the middle stream of Ingoda River. Baikal-Lena grayling has been found for the first time in the headstream of Ingoda River. New data about the distribution of this species in the basin of upper stream of Zeya River are reported; particularly, Baikal-Lena grayling has been found in Okonon Lake; this is the first finding of a representative of Thymallidae family in the alpine lakes of the watershed area of Amur River. In the basins of upper Amur River and Zeya River, the summer ranges of upper Amur grayling Th. grubii grubii and lower Amur grayling overlap to some extent; on the other hand, the ranges of Baikal-Lena grayling and lower Amur grayling are disjoint. During the summer period, upper Amur grayling and Baikal-Lena grayling together inhabit the basin of upper stream of Zeya River, but only Baikal-Lena grayling has been found in the headstream of Zeya River. In the basin of upper Amur River (the headstream of Ingoda River), the joint range of these species is still unknown; the sympatric zone is located in Dzhermoltai River (the basin of Onon River).     

New data on the distribution of the Upper Lena form of grayling (Thymallidae) in the basin of Lake Baikal and its taxonomic status

The habitation of the Upper Lena form of grayling in the upper reaches of the Tiya (the northwestern tributary of Lake Baikal) and Barguzin rivers, where Baikal grayling Thymallus baicalensis Dyb. also occurs, was established. The listed forms can be diagnosed from the body coloration, dorsal fin pattern, combination of meristic characters, as well as by molecular-genetic methods. The sympatric habitation of the Upper Lena and Baikal graylings in the northern tributaries of Baikal, considerable phenotypic and genetic differences, as well as the absence of individuals with intermediate characters, make it possible to consider them as independent species. The results indicate the need to revise the intraspecies structure of the Arctic grayling T. arcticus (Pall.) and the entire genus Thymallus. The localization of populations of the Upper Lena grayling in the upper reaches of Baikal tributaries testifies to its possible penetration of the Baikal basin by means of temporary river catchments that formed in the past and which are probable at present at watersheds with the Lena River. A wider distribution of this form is apparently hindered by the pressure of aboriginal species.     

Selective Feeding of age-0 Arctic Grayling in Lake-Outlet Streams of the Northwest Territories, Canada

A paucity of information exists on the diet of Arctic grayling, Thymallus arcticus, particularly for young-of-the-year (YOY). We examined the diet of YOY Arctic grayling in relation to food availability, in the Barrenlands region of the Northwest Territories, Canada, where lake-outlet streams serve as nursery habitat for these fish. Given the small size of YOY grayling and the abundance of lake-derived microcrustacea in the drift of these lake-outlet streams, we anticipated that these prey would make up a major component of the YOY's diet. Food selectivity by YOY grayling, however, was strongly sized-biased; although microcrustacea dominated the drift, YOY primarily consumed larger taxa, especially Chironomidae and Simuliidae. Even among these taxa, grayling tended to select the larger individuals. As they grew, YOY grayling took larger numbers of both large and small prey, particularly the larger invertebrates, although prey size range did not change after mid-July. Selection of pupae and avoidance of Ephemeroptera suggest that prey characteristics other than size also contribute to selectivity by YOY grayling. The relatively limited consumption of terrestrial invertebrates and other large prey may reflect the small sizes of fish in this arctic study, as well as differences in prey availability. Despite the abundance of lake-derived prey, instream production of invertebrates should largely determine the productive capacity of Barrenlands streams as fish habitat.     

Geographical Variation of Allele Frequencies at the LDH-A* Locus in the Arctic Grayling Thymallus arcticus Pall. and in the European Grayling Thymallus thymallus L.

The allele frequencies of LDH-A* locus were studied in the populations of Siberian grayling from the Kozhym River (Pechora basin) and in the population of European grayling from Pechora, Mezen', and Vym' rivers (Northern Dvina basin). In samples of both species (n = 134), three LDH-A phenotypes have been identified in total, which proved to be under the control of two alleles: LDH-A*100 and LDH-A*50. The alternative alleles of LDH-A* locus were identified in the populations of Siberian grayling from Kozhym River and in the population of European grayling from the same river and other Pechora tributaries, namely, LDH-A*100 and LDH-A*50 in the Siberian and the European grayling, respectively. However, in the European grayling populations from the Mezen' and Vym' rivers, both alleles occur at the frequencies of the rare LDH-A*100 allele of 0.143 and 0.222, respectively. According to the published data, the frequency of LDH-A*100 allele increases in the European grayling populations of northwestern (Finland) and southern Europe (France) rivers, reaching 0.872 and 1.000 in Rhone and Loire, respectively, i.e., the values characteristic of the Siberian grayling populations.     

Finding a new form of the grayling Thymallus arcticus (thymallidae) in the basin of lake baikal

A dwarf form of the Arctic grayling Thymallus arcticus, inhabiting a group of small lakes at the origin of the Yakchii stream, which flows into the Verkhnyaya Angara River (basin of Lake Baikal), has been found in the northeastern watershed of the Baikal and Lena basins. The form is similar to upper Lena populations in body coloration and the pattern of dorsal fin. Our comparison by meristic characters indicated that the grayling of the Yakchii Lakes is more similar to the fish from Kutima River (basin of the upper courses of the Lena River) than to the black Baikal grayling T. arcticus baicalensis. The presence in Lake Baikal of a population similar to upper Lena graylings may have three causes: (1) possible drainage from Baikal to the pra-Lena via ancient valleys of the Barguzin and the Upper Angara; (2) appearance of transit zones as a result of glacial or tectonic events during the periods of watershed development between the Baikal and Lena basins; (3) the upper Lena grayling could be an endemic of the Baikal basin which was replaced by graylings penetrating from the Yenisei basin and remained in the form of relic populations in the upper courses of certain tributaries of the northern and northeastern parts of Lake Baikal.     

Biological specific features of the Lower Amur grayling Thymallus tugarinae (Salmoniformes: Thymallidae): 1. Age composition

We consider age composition of samples of the Lower Amur grayling Thymallus tugarinae inhabiting water bodies of the lower and middle course of the Amur and several rivers of the northwestern part of Sakhalin Island and the southern coast of the Sea of Okhotsk. Age composition of the Lower Amur grayling from the studied samples is not similar because of several reasons. In rivers of the Amur basin, the species has a maximal age of 6+.     

Characterization of 12 highly variable tetranucleotide microsatellite loci for Arctic grayling (Thymallus arcticus) and cross amplification in other Thymallus species

We describe the isolation and development of 12 polymorphic tetranucleotide microsatellite loci for Arctic grayling (Thymallus arcticus). In a sample of 46 fish, we observed between three and 20 alleles per locus. All 12 of these loci were also polymorphic in at least one of the following Thymallus species and subspecies: T. burejensis, T. amurensis, T. thymallus, T. brevirostris, T. grubii, T. arcticus baicalensis and T. arcticus pallasi. These loci will aid in our understanding of the population genetics, behaviour and conservation of grayling species throughout the northern hemisphere.     

Graylings (Thymallidae) of water bodies in western Mongolia: Morphological and genetic diversity

Morphological characters, the pattern on the dorsal fin, some biological parameters, and variations of mtDNA control region were studied in graylings of the upper reaches of the Khovd River (western Mongolia). Obtained results demonstrated that grayling of this part of the river was presented by great (predatory) and small (benthophagous) forms of Mongolian grayling Thymallus brevirostris. The last form was erroneously equated to Arctic grayling Thymallus arcticus. However, genetic unity of great and small forms, homogeneity by meristic characters, and similarity in varying the color of scales and pattern on the dorsal fin attest their belonging to the same species. Water bodies of the Central Asian basin are also populated with populations of Mongolian grayling with mixed type of feeding and signs of external structure, which are typical for both forms in a different degree.     

A new species of grayling <Emphasis Type="Italic">Thymallus svetovidovi</Emphasis> sp. nova (Thymallidae) from the Yenisei basin and its position in the genus <Emphasis Type="Italic">Thymallus</Emphasis>

The Upper Yenisei grayling Thymallus svetovidovi sp. nova that inhabits the headwaters of the Yenisei River in Mongolia is described. From the other representatives of the genus Thymallus, the species differs in the elements of body coloration, dorsal fin pattern, some morphometric characters, and genetic characteristics. Besides this species, most of the Yenisei basin is inhabited by Baikal grayling T. baicalensis Dyb., and its low reaches, by the Arctic grayling T. arcticus (Pall.).     

The trophic interactions of young Arctic grayling (Thymallus arcticus) in an Arctic tundra stream

1. Young (0+) Arctic grayling (Thymallus arcticus) have the potential to control the trophic structure of Arctic tundra streams through consumption, nutrient excretion and the modification of prey behaviour. The effect of young grayling on three trophic levels (algae, invertebrates and fish) was investigated by manipulating fish density and by fertilizing the river with phosphorus (P). 2. Nutrients, epilithic chlorophyll a, benthic invertebrates and fish biomass were measured within each fish density treatment (0, 4, and 40 fish m^–2) within the P-limited reference zone and the P-enriched fertilized zone of the Kuparuk River, Alaska. 3. Epilithic chlorophyll a increased with increased fish density in both reference and fertilized zones, while mayfly density decreased with increased fish density in the fertilized zone only. Final mean mass of young grayling in the 40 fish m^–2 cages was lower than mean mass in the 4 fish m^–2 cages. 4. Young grayling may produce a top-down cascading trophic effect in areas where nutrients are not limited. 5. River nutrient status and river discharge may modify the strength of top-down control by young grayling.     

Age and growth comparisons of Hovsgol grayling (Thymallus nigrescens Dorogostaisky, 1923), Baikal grayling (T. baicalensis Dybowski, 1874), and lenok (Brachymystax lenok Pallas, 1773) in lentic and lotic habitats of Northern Mongolia

Despite concern over the conservation status of many Mongolian salmonids and the importance of their ecological role in Mongolia's aquatic ecosystems, little is known about their basic biology. Hovsgol grayling (Thymallus nigrescens) is endemic to Lake Hovsgol, Mongolia and listed as endangered on the Mongolian Red List. Baikal grayling (T. baicalensis) and lenok (Brachymystax lenok) are found in lakes and rivers throughout the Selenge drainage. A detailed study of the age and growth of these three salmonids was conducted based on 1,682 samples collected from July 2006 to July 2013 in Lake Hovsgol, its outlet the Eg River, and one of the Eg's largest tributaries, the Uur River. Our results suggest that Hovsgol grayling in particular can reach a much older maximum age (17 years in our samples) than previously believed based on aging from scales. Female Hovsgol grayling were heavier at a given length than their male counterparts. Lenok had a greater average length‐at‐age in Lake Hovsgol compared to the rivers and greater weight‐at‐length in the warmer Uur River than in the Eg; female lenok from the rivers had a greater average length‐at‐age than their male counterparts. This study provides critical new information for the management and conservation of these threatened salmonid species in Mongolia.     

The impact of stocking on the genetic structure of European grayling <Emphasis Type="Italic">Thymallus thymallus</Emphasis>, Salmonidae) in two alpine rivers

The European grayling (Thymallus thymallus) is considered to be threatened in several European regions. In recent decades fishery managers have increasingly turned to stocking programs as one way to combat the negative effect that human influence has had on population densities. The present study surveyed the genetic structure of two Danube drainage populations at the Inn and Drau rivers, in Austria, on the basis of mtDNA sequences. Data were placed in the context of the phylogeographic structure of European Thymallus and thus could reveal unexpected geographical mixing due to stocking with allochthonous individuals. Our analyses revealed that regular stocking of fish not originating from their natal Rivers has left genetic traces in both systems surveyed. These traces may be classified as marginal for the Inn river and its tributaries in which 97% of the graylings investigated carried haplotypes belonging to the northern alpine lineage, corresponding to the region through which the Inn flows. In contrast, the genetic composition of the Drau population, situated in the southern Alps, has been seriously altered through the stocking of fish belonging to the northern alpine mtDNA lineage as only 62% of the fishes sampled carried haplotypes representing the native southern alpine lineage.     

Aggression Level in Different Water Velocities Depends on Population Origin in Grayling, Thymallus thymallus

Phenotypic plasticity is recognized as an important mechanism of adaptation. However, because of its potential costs and limits, it has been hypothesized to be reduced and ultimately become lost when there is no selection for its maintenance. Heterogeneous environments in particular are expected to favour and thus maintain plastic phenotypes. Lakes and rivers differ in their flow regimes. In addition to higher average water velocity, rivers are characterized by distinct spatial and temporal variation in water flow, whereas lakes can be regarded as quite uniform in this respect. We studied whether behaviour, which is generally considered to be highly plastic, shows differences in the degree of plasticity between lake and river populations of the European grayling, Thymallus thymallus, fry in response to different water velocities. Given that aggressive behaviour in fish has been shown to relate to ambient flow regime, we compared aggressiveness of hatchery‐reared grayling originating from hatchery stocks of two lake and two river populations in still and flowing water. River fish showed higher aggressiveness in flowing water compared with still water, whereas aggressiveness of lake fish did not appear to vary according to water velocity. The higher plasticity of aggressiveness evoked in river fish by different water velocities may thus represent an adaptation to more variable flow and presumably related feeding conditions in their natural environment.     

New species of grayling <Emphasis Type="Italic">Thymallus tugarinae</Emphasis> sp. nova (Thymallidae) from the Amur River Basin

Data on a new species, low Amur grayling Thymallus tugarinae sp. nova, inhabiting tributaries of the lower and middle current of the Amur River, are presented. This species has been earlier equated with the Amur grayling T. grubii, described by Dybowski (1869) from the rivers Onon and Ingoda (the Upper Amur Basin). The new species differs from other representatives of the genus in the body coloration, the pattern on the upper fin, and certain morphological characters. On most of the Amur Basin, the grayling from the Lower Amur is sympatric with T. grubii and in its tributary Bureya River, also with the Bureya grayling T. burejensis. Individuals with intermediate traits are unknown, pointing to reproductive isolation. These data are supported by the results of molecular-genetic analysis (Froufe et al., 2003, 2005; Knizhin et al., 2004).     

Phenotypic and genetic differentiation of two major phylogeographical lineages of arctic grayling Thymallus arcticus in the Lena River, and surrounding Arctic drainages

Two phylogeographical lineages of arctic grayling, Thymallus arcticus , in Siberia are extensively characterized based on both molecular genetic (mtDNA control region sequences) and phenotypic (12 meristic characters) data. One lineage, occurring in the delta region of the Lena River as well as all other Arctic draining rivers sampled, corresponds to the subspecific taxon Thymallus arcticus pallasii , whose type locality is the Kolyma River. This taxon is proposed to be a postglacial colonizer of the Lena delta. The second lineage occurs throughout the rest of the Lena basin and is proposed to have survived in a glacial refugium in the middle reaches of the Lena. These lineages form reciprocally monophyletic groups based on mtDNA sequences (net divergence 3.2%), a relationship that is concordant with phenotypic data, and thus reflects distinct taxa. The upper Lena taxon is given the preliminary name of Thymallus arcticus lenensis . Phylogenetic analysis, together with previously published data from North America, reveals that mtDNA sequences from North American populations group within the diverse clade corresponding to T. a. pallasii in Siberia. Despite the relatively close genetic relationship of most North American haplotypes with those in northern Siberia, inferences of fragmentation between the continents are supported, but bidirectional movements between the two continents are seen as likely. Despite inclusion in the clade representing T. a. pallasii in Siberia, the source of the relatively divergent Nahanni refuge haplotypes in North America is not resolved. Otherwise, inferences of postglacial expansion across several thousand kilometres are well supported within North America, but only smaller-scale colonization events among drainages are supported in Siberia.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 88 , 511–525.