A new subspecies of the amur grayling Thymallus grubii flavomaculatus ssp. nova (Thymallidae)

A new subspecies of the Amur grayling is described, the yellow-spotted grayling Thymallus grubii flavomaculatus ssp. n. populating the upper reaches of large tributaries of the Lower Amur basin and some rivers flowing to the Tatar Strait, the Sea of Okhotsk, and the Sea of Japan. From nominative subspecies T. g. grubii populating the basin in the upper reaches of the Amur and from other representatives of the genus Thymallus, this subspecies differs in the dorsal fin pattern whose principal trait is a yellow spot on the last two-five interray membranes, in the body coloration, and in some morphological characters. The yellow-spotted grayling in the upper reaches of tributaries sympatrically coexists with the Lower Amur grayling Thymallus sp. No specimens with intermediate characters are found. The sympatric forms spawn at different times, thus being isolated reproductively. Isolation of the yellow-spotted grayling as a subspecies of the Amur grayling T. grubii is confirmed by the results of molecular-genetic investigations.     

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).     

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).     

Biological features of Low Amur grayling Thymallus tugarinae (Salmoniformes: Thymallidae): 2. Growth

We present the results of analysis of the group growth of the Low Amur grayling Thymallus tugarinae from 18 rivers of the Low Amur basin, tributaries of the Bureya and Ussuri rivers. We made use of the published data on the fishes of Sakhalin Island. The dwarf and fast-growing forms are not revealed among the investigated groups of this species. The parameters of growth of the Low Amur grayling in the investigated rivers are different, which is determined by the different conditions of feeding migration connected with the geographical location of the streams, their hydrological features, density of populations, etc. The anthropogenic factor influence is essential. Growth rate of T. tugarinae is lower compared with the other Amur species of graylings.     

Biological characteristics of lower Amur grayling Thymallus tugarinae (Salmoniformes: Thymallidae): 3. reproductive biology

We discuss peculiarities of the reproductive biology of lower Amur grayling Thymallus tugarinae. The sex ratio in most of the studied samples was close to 1 : 1, but sometimes it might differ due to various reasons. Females dominated among the specimens entering the first reproduction period; the rate of females decreased in the fish older than 5 years. Maturation was usual in fish that reached 20-cm body length, which referred to the age 3+. The males became mature a year after the females of the same age in a half of the studied rivers. Absolute fecundity, egg size and weight, as well as gonad-somatic index, correlated positively to the age of the fish, its body weight, and length. The lower Amur grayling spawned from the end of April through the last decade of May at the water temperatures exceeding +6°C in the downwelling areas located upstream the riffles, on the small-size gravel bottom sediments, at the depths of 0.8 m and less, where the current velocity was less than 0.7 m/s. The spawning grounds in the main riverbed and tributaries of the lower stream of the Anyui River comprised 426.3 ± 148.6 m² per km², or 0.32 ± 0.11% of the water-surface area.     

A comparative analysis of two Amur grayling species from the <Emphasis Type="Italic">Thymallus</Emphasis> Genus (Salmoniformes; Thymallidae) according to a number of osteological and morphological features in their zone of sympatry

The results of the comparison of the Lower Amur grayling Thymallus tugarinae and the yellowspotted grayling T. flavomaculatus that were collected in the zone of their sympatry in the middle reaches of the Anuy River (the Lower Amur basin) for a number of osteological and morphological features are provided. The work was based on the analysis of X-ray images and a number of plastic features. Considerable differences were detected between the two species in the numbers of the bone elements of the axial skeleton. The differences in the number and topography of certain bone elements correlate with the morphological features of these species. The comparison results for certain features do not correspond with the literature data.     

Seismosensory system of the Lower Amur rayling <Emphasis Type="Italic">Thymallus tugarinae</Emphasis> (Thymallidae) from the Anyui River

Data on specific features of the seismosensory system of the Lower Amur grayling Thymallus tugarinae from the Anyui River (basin of the Lower Amur) are provided. According to the number of openings in canals of the seismosensory system, individuals from the middle course significantly differ from graylings from the lower course in greater values of interpopulation diversity and fluctuating asymmetry. According to the number of pores of canals of the seismosensory system, the Lower Amur grayling differs from the East Siberian T. arcticus pallasi, Kamchatka T. a. mertensii, and Alaska T. a. signifer graylings.     

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.     

On the diversity of fishes in mountain lakes of the Amur basin

Results of the study of 11 lakes situated in the mountainous part of the Amur River basin are reported. Six fish species, namely, blunt-snouted lenok Brachymystax tumensis, Baikal-Lena grayling Thymallus baicalolenensis, Siberian stone loach Barbatula toni, Lagowski’s minnow Rhynchocypris lagowskii, Czekanowski’s minnow Rh. czekanowskii, and Amur sculpin Cottus szanaga, were found in five lakes. Four species were found in one lake (Lake Pereval’noe, basin of the Amgun’ River), and two species were found in each of the remaining lakes. Lenok and Amur sculpin were the most common species. The fauna of Lake Okonon (basin of the river Zeya) was the most specific and did not have analogs among the lake faunas; Czekanowski’s minnow and Baikal black grayling were found in this lake. The lenok ecotype that formed in the mountain lakes differed from the river lenok in some regards. The potential pathways of formation and protection of mountain lake ichthyocenoses of the Amur basin are discussed.     

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.     

Distribution,abundance, and size structure of Amur kaluga <Emphasis Type="Italic">Acipenser dauricus</Emphasis> and Amur sturgeon <Emphasis Type="Italic">A. schrenckii</Emphasis> in the Lower Amur and Amur Estuary

Distribution, population structure, and abundance of Amur kaluga Acipenser dauricus and Amur sturgeon A. schrenckii in the Lower Amur and Amur Estuary have been considered based on the materials of two surveys performed in 2011. Both species occur along almost the entire length of the Lower Amur and in major part of the Amur Estuary. The density of their aggregations has significantly decreased in most regions of the river if compared to the 1960s. In the Amur Estuary, A. dauricus and A. schrenckii are most abundant in freshwater and light salted water bodies of its western part. In 2011, the abundance and biomass of A. dauricus and A. schrenckii in the Lower Amur and Amur Estuary was 345000 specimens and 7110 tons and 289000 specimens and 1946 tons, respectively.     

Biological and morphological characteristic of the Arctic grayling Thymallus arcticus (Thymallidae) from Alpine Lakes of the basin of the upper reaches of the Angara River

Specific morphological and biological features of the populations of the Arctic grayling Thymallus arcticus from lakes Urunge Nur, Mongosha, Sadaiskoe, and Gusinoe situated in the Upper Sayan Mountains in the sources of the Oka and Kitoi rivers (the basin of the upper reaches of Angara) that have been not studied previously are considered. The results of the multivariate analysis indicated the uniformity of all samples by 12 meristic characters. Graylings of the studied lakes are similar to graylings from the Angara and Nizhnyaya Tunguska rivers and the Irkutsk Reservoir, as well as to the Baikal grayling T. a. baicalensis in body coloration and pattern of the dorsal fin. The samples studied and T. a. baicalensis considerably differ in the above characters from populations inhabiting the basin of the upper reaches of the Ob that belong to the nominative subspecies T. a. arcticus. The differences established between the Angara-Yenisei and the Upper Ob graylings give grounds to doubt the justification of including them into one taxon, T. a. arcticus. The same is evidenced by the results of molecular-genetic studies of graylings from the Palearctic (Froufe et al., 2005). The data obtained indicate the necessity of ascertaining the boundaries of the ranges of different forms of graylings in the Angara-Yenisei and Ob basins and of the revision of their taxonomic status. Populations inhabiting the Angara and Yenisei rivers, except their sources and the section of the lower reaches, should be assigned to the Baikal subspecies, T. a. baicalensis. Graylings from different lakes in the upper reaches of Oka and Kitoi differ in their linear-weight indices. The indices of growth are highest where they occupy a dominant position. Their food in the summer consists mainly of the larval and imaginal stages of amphibiotic and aerial-terrestrial insects. Only in Lake Urunge Nur do graylings of elder age groups use fish of other species for food. The extreme conditions of habitation of the grayling in Lake Gol’tsovoe are the cause of food deficiency, which determines its dwarfism.     

Identification of reproductively isolated lineages of Amur grayling (Thymallus grubii Dybowski 1869): concordance between phenotypic and genetic variation

We analysed variation at maternally (mitochondrial DNA control region sequences) and bi-parentally (10 microsatellites) inherited genetic markers, as well as across 12 meristic characters in 7 populations of Amur grayling, Thymallus grubii, from eastern Siberia. All three data sets were concordant in supporting the existence of three diagnosable, reciprocally monophyletic, and most probably reproductively isolated, lineages of grayling within the Amur drainage. There was a significant correlation between genetic and phenotypic divergence, both within and among lineages. Two phenotypically distinct forms (with and without an orange spot on the posterior portion of the dorsal fin), found in sympatry in the lower Amur, most likely result from secondary contact, as they demonstrate 4.6% sequence divergence at the mitochondrial DNA control region. This divergence, together with the existence of at least one nearby population of orange spot grayling outside the Amur drainage (0.8% divergence) underscore the palaeo-hydrological complexity of the system, which presumably promoted genetic divergence in a shifting allopatric framework throughout the Pleistocene. Grayling from the upper Amur, corresponding to the type locality for the species, formed a sister group (1.4-1.6% divergent) to the orange spot lineage perhaps diverging in the early Pleistocene (1.4-1.6 Ma).     

Distribution of Masu Salmon <Emphasis Type="Italic">Oncorhynchus masou</Emphasis> (Salmonidae) in Amur River Basin

The presented data on the distribution of the masu salmon Oncorhynchus masou in the Amur River Basin are collected in 1976–2015. In the analyzed region the species’ spawning range includes the lower part of the Amur River basin from the rivers flowing into the Amur estuary to the basins of the Anyui and Amgun’ Rivers in their middle courses. The information on the catch of a masu salmon specimen in the Khor River Basin (approximately 1300 km from the Amur River mouth) in 2015 is presented. It is assumed that the total Amur population does not exceed several thousand of adult anadromous fish.     

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.     

Geographical variation of allele frequencies at the LDH-A* locus in the arctic grayling Thymallus arcticus Pall. and in the European grayling Thymallus arcticus L

The allele frequencies of LDH-A* locus were studied in the population 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 (France) rivers, reaching 0.872 and 1.000 in Rhone and Loire, respectively, i.e., the values characteristic of the Siberian grayling populations.     

Endemic sturgeons of the Amur River: kaluga, Huso dauricus, and Amur sturgeon, Acipenser schrenckii

General biological characteristics and the contemporary status of the kaluga, Huso dauricus, and Amur sturgeon, Acipenser schrenckii, are described. Both inhabit the Amur River basin. Kaluga is the largest freshwater fish in this river system reaching more than 5.6 m in length and more than 1000 kg in weight. We recognize four populations of kaluga: the first is from the estuary of the Amur River and coastal brackish waters of the Sea of Okhotsk and Sea of Japan, the second is from the lower Amur River, the third is from the middle-Amur, and the fourth occurs in lower reaches of the Zeya and Bureya rivers. Freshwater and brackish water morphs exist in the estuary population, with the freshwater morph predominating in number. The number of individuals in the lower Amur River population at age 2 or greater was recently estimated to be 40 000, and in the middle Amur, 30 000. The population will continue to decline because of rampant overfishing. The Amur sturgeon is represented in the Amur River basin by two morphs: brown and gray. Brown morphs occur in the middle and lower parts of the Amur River; they grow more slowly than the gray ones. Today, the lower Amur River population of Amur sturgeon is made up of 95 000 fish at age 2 or greater and is approximately half as large as the population in the middle Amur River. Populations of kaluga and Amur sturgeon in the Zeya and Bureya rivers are extremely small and on the verge of extinction.     

Mitochondrial and nuclear DNA phylogeography of Thymallus spp (grayling) provides evidence of ice-age mediated environmental perturbations in the world's oldest body of fresh water,Lake Baikal

Theories on the hydrological history of Lake Baikal, the world's oldest and deepest body of freshwater, and its surrounding great rivers, are currently based solely on geological evidence and are conflicting. Baikal is inhabited by numerous zoogeographical enigmas but their high level of endemism has hindered phylogeographic inferences. We provide a biological perspective of the region's palaeo-hydrological development based on the demographic and genealogical history of the widespread Thymallus spp. (grayling). Phylogenetic reconstruction reveals that old lineages of grayling (pre-Pleistocene) currently inhabit the Enisey, Lena and Amur River basins. For Lake Baikal however, we conclude that a mid-Pleistocene colonization (110000-450000 years ago) of an unoccupied niche has occurred. Population genetic inferences support an Enisey-Angara river route of colonization into Baikal, corresponding to the cataclysmic palaeo-hydrological event that led to the formation of the lake's only contemporary outlet, and a subsequent range expansion several thousand kilometres into the uppermost reaches of the Selenga River basin. The evolutionary history of Lake Baikal grayling is congruent with the controversial hypothesis of repeated glaciation. However, considering the extraordinary levels of endemism and proposed Miocene or Oligocene coalescence of other faunal lineages, a less profound but equally consequential cycle of environmental perturbations may have taken place. Bi-parentally inherited microsatellite DNA loci supported the phylogenetic relationships of Thymallus spp. and the geographical expansion of Baikal grayling strikingly well. A Markov Chain Monte Carlo modelling approach suggested severe contemporary population decline during the last century, possibly reflecting the influence of an uncontrolled fishery on this treasured ecosystem. These complementary pictures of the demographic history of grayling underscore the breadth of historical inquiry that can be entertained through the modelling of sufficient molecular data, and may significantly alter the zoogeographical and limnological perspectives of Baikal's history.     

Age and growth of kaluga Acipenser dauricus from the estuary of the Amur and its lagoon

Age composition and the rate of growth of kaluga Acipenser dauricus from the estuary of the Amur and the Amur lagoon have been analyzed. Differences in the distribution of juvenile and mature individuals of kaluga in the river estuary and the lagoon were revealed. Males are dominant among mature individuals aged 18–26 years, and females are dominanat at an age 27–43 years. Females and males of the same age do not differ in length and body weight. A decrease in the rate of linear and gravimetric growth of mature individuals at the modern stage (2005–2008) compared to the period of 1965–1982 was revealed. Juvenile individuals of kaluga from the near-estuarine part of the Amur and its lagoon are inferior in the rate of growth to fish of the same age caught in the river channel, 500 km to the south.     

Divergence of the seasonal races of chum salmon, <Emphasis Type="Italic">Oncorhynchus keta</Emphasis> Walbaum, 1792, in the Amur and Poronai rivers: Ecology,genetics, and morphology

The chum salmon of the Amur River (the mainland part of the Far East) and the Poronai River (Terpeniya Bay, Sakhalin Island) are historically related to one another, as the drainage basins of these rivers are the remnants of a formerly single river system, the Paleoamur, which existed when Sakhalin Island was a part of the continent. Both river populations of chum salmon consist of the early-run and late-run ecological forms (seasonal races), which are also referred to as the summer and autumn races. They are reproductively isolated from each other due to their spawning at different times and in different types of spawning grounds. To assess the direction, pattern, and degree of divergence between these chum salmon races in the both river fragments since the Paleoamur, it is necessary to compare them using two types of traits: selectively neutral DNA markers and morphological and physiological traits, variations in which may have an adaptive value. For this, we have studied chum salmon from both rivers in terms of microsatellite DNA markers, body counts and measurements, body weight, and fecundity. Both in the Amur River and in the Poronai River, the autumn race of chum salmon prevails over the summer race in body length and weight, fecundity, number of pyloric caeca, and several other meristic traits. The intra-basin differences between the races are much more pronounced in the Amur chum salmon. The inter-race differences in microsatellites are also greater in the Amur chum salmon compared to the Poronai chum salmon. Using microsatellites, three levels of differentiation have been revealed: (1) between the basins of the Amur and Poronai rivers, (2) between the races within each of the river basins, (3) and between population samples within each race of each basin. A hypothesis is proposed that the currently existing races of chum salmon in the Amur and Poronai rivers have evolved since the breakup of the Paleoamur, and the intra-basin divergence of the races started in the Amur River earlier than in the Poronai River. An analysis of our own data and the published data suggests that the adaptation of the seasonal races of chum salmon to the conditions of their spawning grounds is determined by a complex of morphological and physiological traits, including the number of pyloric caeca, which is an adaptive and highly heritable trait associated with the incubation temperature of the water.