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+.     

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).     

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.     

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.     

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.     

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.     

A comparative analysis of the Far Eastern grayling species Thymallus tugarinae and Thymallus grubii flavomaculatus based on the data from mitochondrial DNA cytochrome b gene sequencing

Nucleotide sequences of a fragment of mitochondrial DNA cytochrome b gene were obtained from two species of graylings that inhabit the Russian Far East, viz., Thymallus tugarinae and T. grubii flavomaculatus. A phylogenetic analysis of four Thymallus species, whose relationships have been poorly studied, was performed on these new data as well as on several sequences from the NCBI GenBank sequence database. The rate of genetic divergence between the Far Eastern graylings Thymallus tugarinae and T. g. flavomaculatus corresponded to that between European (T. thymallus) and Arctic (T. arcticus) graylings. Moreover, single nucleotide substitutions that result in alterations of the amino-acid structure of protein products (non-synonymic mutations) were revealed between sequences of the cytochrome b gene in the Far Eastern grayling species. The topology of phylogenetic trees, which was composed by means of Bayesian analysis and the maximum parsimony method, showed four independent phylogenetic lineages of graylings. In addition, the phylogenetic relationships between T. tugarinae and T. g. flavomaculatus were supported statistically. The obtained data indicated that Far Eastern graylings T. tugarinae and T. g. flavomaculatus are distinct species and confirmed that they belong to different phylogenetic lineages, as found earlier.     

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.     

Comparison of biology of the Sakhalin sturgeon, Amur sturgeon, and kaluga from the Amur River, Sea of Okhotsk, and Sea of Japan biogeographic Province

Three Acipenseridae species live in the rivers and marine waters of the Khabarovsk Territory, Russia: Sakhalin sturgeon, Acipenser mikadoi, Amur sturgeon, A. schrenckii, and kaluga, Huso dauricus. We review the general biology and life history of each species, including their historic and current distributions, and examine current paleogeographic theories to outline the possible origin and evolution of these three species in the Amur River, Sea of Okhotsk, and Sea of Japan biogeographic province. Apparently, these species have evolved during distinct geologic time periods, which has reinforced the reproductive isolation of these species although hybridization does occur. They have convergently adapted to the unique environmental conditions of the Amur River and Russian Maritime regions, and yet developed behavioral adaptations to reduce competition between species. Sakhalin sturgeon is the least studied species among anadromous sturgeon in the world. This species is highly migratory and spends the majority of its life in the ocean only returning to natal rivers to spawn. Amur sturgeon and kaluga are distributed throughout the Amur River basin and the estuary and share many life history traits. They are both represented by distinct morphs. Additionally, we present size and weight relationships to estimate the growth of Amur sturgeon and kaluga. All three species have suffered declines in abundance due to over fishing and their contemporary distributions have contracted compared to their historic ranges. We identify gaps in knowledge and suggest further research useful for guiding management of each species.     

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.     

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.     

Comparative biology and management of Arctic and European grayling (Salmonidae,Thymallus)

Summary The biological and ecological features of Arctic grayling (Thymallus arcticus) relevant to its management and enhancement are reviewed over its range in North America as well as in Asia, and are compared with those for the European grayling (Thymallus thymallus). Populations of both species have declined in some areas where angling pressure has been severe and where major alterations of their habitats have occurred. But native populations of Arctic grayling have been extirpated in eastern parts of its North American range, and nearly so in south-western parts, whereas the range of European grayling has been extended, especially in the United Kingdom and in Finland, by introduction. Furthermore, in contrast to the Arctic grayling, the European grayling, in Scandinavia at least, seems to have been less severely affected by impoundment of rivers and lakes for hydroelectric generation, perhaps because it has been able to successfully reproduce in large rivers below dams, especially those where fluctuation in water level is small.The success of various grayling management and enhancement practices, including hatchery culture and stocking, angling restrictions, spawning and rearing facilities, competitor and predator control, stream habitat restoration, lake and stream fertilization, and invertebrate prey introduction, are evaluated and compared for both species.     

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.     

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.     

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).     

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 influence of the invasive shrub, Lonicera maackii, on leaf decomposition and microbial community dynamics

Amur honeysuckle (Lonicera maackii) is an exotic invasive shrub that is rapidly expanding into forests of eastern North America. This species forms a dense forest understory, alters tree regeneration, negatively affects herb-layer biodiversity, and alters ecosystem function. In a second-growth forest in central Kentucky, we examined the timing and production of leaf litter and compared litter chemistry, decay rates, and microbial community colonization of Amur honeysuckle to that of two native trees, white ash (Fraxinus americana) and hickory (Carya spp.). The distribution of Amur honeysuckle was clumped, allowing us to compare differences in decomposition under and away from Amur honeysuckle shrubs. Amur honeysuckle leaf litter had significantly higher nitrogen, lower C:N, and lower lignin than the other species, and decomposition rates were greater than 5×?faster. Despite the much higher rate of Amur honeysuckle decomposition compared with the native species (p?<?0.0001), decomposition of all species was significantly slower (p?=?0.0489) in sites located under Amur honeysuckle shrubs. Nitrogen concentration increased through time in decomposing Amur honeysuckle litter; however, total mass of N rapidly declined. We found the initial microbial community on leaf litter of Amur honeysuckle was distinct from two native species and although all microbial communities changed through time, the microbial community of Amur honeysuckle remained distinct from native communities. In summary, a distinct microbial community that may originate on Amur honeysuckle leaves prior to senescence could explain the rapid decay rates.