The developmental cycle of Proteocephalus thymalli (Cestoda, Proteocephalidae)--a parasite of the Arctic grayling from Lake Baikal

The life cycle of Proteocephalus thymalli (Cestoda, Proteocephalidae), a parasite of Siberian glame (Thymallus arcticus), was repeated under experimental conditions. The first intermediate hosts, the copepods Epischura baicalensis, Cyclops kolensis and C. vicinus, were determined. The developmental time of P. thymalli in the first intermediate host was determined and the morphology of the larval and adult phases was described.     

Description of a new species of Proteocephalus pronini sp.n. (Cestoda: proteocephalidae)--a parasite of grayling from lake Hovsgol

A new cestode species, Proteocephalus pronini sp. n., is described from the Lake Hovsgol Grayling Thymallus arcticus nigrescens (Mongolia, Hovsgol Lake). The new species differs from P. thymalli from the Baikal Lake by the length of strobile, number of proglottids, form of mature proglottids, and number of diverticles of uterus.     

北极茴鱼胚胎及仔鱼发育

研究对人工繁殖的北极茴鱼(Thymallus arcticus grubei)胚胎发育开展系统观察,记录分析其胚胎及仔鱼发育各时期的形态特征,旨在为北极茴鱼的人工繁育和种质资源保护提供必要的基础数据。结果显示,北极茴鱼受精卵呈圆球形,金黄色,沉性卵,未吸水卵径(2.46±0.14) mm,吸水卵径(3.14±0.18) mm,卵黄质内有多个油球。油球的数量和空间分布在胚胎发育过程中发生了规律性变化。在孵化水温(11.06±0.72)℃,溶氧8.3—9.8 mg/L条件下,历时301h完成整个胚胎发育过程,所需积温为3384.84h·℃,经历合子期、卵裂期、囊胚期、原肠胚期、神经胚期、器官形成期和孵化出膜7个阶段,共26个时期。北极茴鱼仔鱼尾鳍、胸鳍的分化和眼色素沉积在受精卵胚胎发育后期就已经完成,背鳍、腹鳍、臀鳍和脂鳍等在胚后发育过程中相继分化。其初孵仔鱼平均全长为(9.33±0.35) mm,仔鱼卵黄囊呈圆球形, 18日龄时卵黄囊和油球被完全消耗。其早期发育阶段(0—16日龄)的生长特性符合公式:y=0.0005x⁴–0.0201x³+0....     

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.     

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.     

Mitochondrial DNA variation of Gyrodactylus spp (Monogenea, Gyrodactylidae) populations infecting Atlantic salmon, grayling, and rainbow trout in Norway and Sweden

Approximately 800 bp of the mitochondrial cytochrome oxidase I (COI) gene were sequenced from 76 Gyrodactylus specimens of 32 salmonid host populations, i.e. from Salmo salar, Thymallus thymallus, and Oncorhynchus mykiss in Norway, Sweden and Latvia. The COI sequences indicated a substantial intraspecific differentiation of Gyrodactylus salaris and Gyrodactylus thymalli. In total, 12 haplotypes were identified which group into five well supported clades, three clades with parasites from Atlantic salmon and two clades with parasites from grayling. The basal nodes linking the five clades together are only weakly supported. Thus, there is no support for the monophyly of all G. salaris haplotypes and the monophyly of all G. thymalli haplotypes. The lack of monophyly of the mitochondrial haplotypes of G. salaris and G. thymalli may indicate that G. salaris and G. thymalli represent (i). two polytypic species or (ii). one polytypic species, or (iii). refer to a complex of more than two sibling species. The mtDNA data indicate multiple introductions of G. salaris and G. thymalli into Norway. A minimum of three independent introductions of G. salaris and two independent introductions of G. thymalli are supported. This is congruent with earlier hypotheses on the introduction of G. salaris and G. thymalli into Norway.     

Cytochemical characteristics of the myxosporidian Henneguya cerebralis (Myxosporidia, Myxobolidae) and pathohistological changes in the Kosogol grayling in myxosporidiasis

Content and distribution of RNA, protein, DNA, glycogen and carbohydrate components in spores and vegetative sporeforming stages plasmodium of Henneguya cerebralis were described. Pathohistological changes in head tissus of Thymallus arcticus nigrescens are discussed. The conclusion on the relative youth of parasite system "H. cerebralis--Thymallus arcticus nigrescens" is made.     

线粒体DNA序列变异显示鸭绿江茴鱼为黑龙江茴鱼同物异名

经测序获得北极茴鱼鸭绿江亚种Thymallus arcticus yaluensis和黑龙江茴鱼T.grubii共26尾的线粒体DNA控制区基因全序列,以下游黑龙江茴鱼T.tugarinae、北极茴鱼指名亚种T.a.arcticus和欧洲茴鱼T.thymallus作外群,构建了茴鱼属部分种分子系统发育树,以探讨北极茴鱼鸭绿江亚种分类地位的有效性。结果显示,鸭绿江茴鱼与黑龙江茴鱼的4个地理种群(呼玛河、甘河、牡丹江、松花江)间的Kimura 2-parameter遗传距离仅为0·007~0·015,处于黑龙江茴鱼不同地理种群间的变异范围之内(0·004~0·015),明显小于属内种间的遗传距离(0·046~0·062)。以邻接法(NJ)和最大简约法(MP)构建的茴鱼属部分种系统发育树表明,鸭绿江茴鱼未构成独立的单系群,也未与北极茴鱼T.a.arcticus形成亚种关系,而是聚到黑龙江茴鱼种群内部,共同构为一个单系群。研究表明,鸭绿江茴鱼T.a.yaluensis并非有效种,应为黑龙江茴鱼T.grubii的同物异名,可视为黑龙江茴鱼的一个地理生态种群。     

Occupancy of young-of-year Arctic grayling (Thymallus arcticus) in Barrenland streams

Hydrobiologia - Arctic grayling (Thymallus arcticus) is an iconic fish species that is present across the remote subarctic Barrenlands, yet our lack of understanding of their distributional...     

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.     

Phylogeographical lineages of Arctic grayling (Thymallus arcticus) in North America: divergence, origins and affinities with Eurasian Thymallus

The number and location of Arctic glacial refugia utilized by taxa during the Pleistocene are continuing uncertainties in Holarctic phylogeography. Arctic grayling (Thymallus arcticus) are widely distributed in freshwaters from the eastern side of Hudson Bay (Canada) west to central Asia. We studied mitochondrial DNA (mtDNA) and microsatellite DNA variation in North American T. arcticus to test for genetic signatures of survival in, and postglacial dispersal from, multiple glacial refugia, and to assess their evolutionary affinities with Eurasian Thymallus. In samples from 32 localities, we resolved 12 mtDNA haplotypes belonging to three assemblages that differed from each other in sequence by between 0.75 and 2.13%: a 'South Beringia' lineage found from western Alaska to northern British Columbia, Canada; a 'North Beringia' lineage found on the north slope of Alaska, the lower Mackenzie River, and to eastern Saskatchewan; and a 'Nahanni' lineage confined to the Nahanni River area of the upper Mackenzie River drainage. Sequence analysis of a portion of the control region indicated monophyly of all North American T. arcticus and their probable origin from eastern Siberian T. arcticus at least 3 Mya. Arctic grayling sampled from 25 localities displayed low allelic diversity and expected heterozygosity (H(E)) across five microsatellite loci (means of 2.1 alleles and 0.27 H(E), respectively) and there were declines in these measures of genetic diversity with distance eastward from the lower Yukon River Valley. Assemblages defined by mtDNA divergences were less apparent at microsatellite loci, but again the Nahanni lineage was the most distinctive. Analysis of molecular variance indicated that between 24% (microsatellite DNA) and 81% (mtDNA) of the variance was attributable to differences among South Beringia, North Beringia and Nahanni lineages. Our data suggest that extant North American Arctic grayling are more diverse phylogeographically than previously suspected and that they consist of at least three major lineages that originated in distinct Pleistocene glacial refugia. T. arcticus probably originated and dispersed from Eurasia to North America in the late to mid-Pliocene, but our data also suggest more recent (mid-late Pleistocene) interactions between lineages across Beringia.     

Finding of the subarctic leeches, Acanthobdella peledina and Cystobranchus mammillatus, in the Lake Baikal basin and the reasons for their absence in Lake Baikal itself]

Acanthobdella peledina was first found in the basin of Lake Baikal on the fluvial form of the Balkal grayling Thymallus arcticus baicalensis and Coregonus lavaretus pidschian from the Upper Angara. Cystobranchus mammillatus, a parasite of burbot, is widely distributed in the tributaries of Baikal. The absence of these subarctic leeches from Baikal itself is explained by their rheophily.     

Zoogeographical implications of variation in mitochondrial DNA of Arctic grayling (Thymallus arcticus)

Most of the northern half of North America's freshwater fish survived the last glacial period in unfrozen refugia peripheral to the ice sheets. In our study, the question of which refugia Arctic grayling ( Thymallus arcticus ) inhabited during the Wisconsinan Ice Age, and how they subsequently dispersed to their present geographical range, was examined using mitochondrial DNA (mtDNA) analysis. mtDNA from 12 T. arcticus populations was analysed by direct sequencing and restriction fragment length polymorphism analysis (RFLP). Our data support the hypotheses that T. arcticus had a large refugial population in the Bering Refuge (shown by high mtDNA diversity in extant Alaskan populations) and that British Columbia was colonized from the Bering Refuge (shown by mtDNA haplotype similarities). Our data also show that a disjunct southern set of populations in Montana is significantly different from the northern grayling, in terms of restriction haplotype frequency and distinguishing sequence characteristics. Sequence results yielded an estimated divergence time of 370 000 years between the northern and Montana grayling haplotypes. We conclude that T. arcticus survived the Wisconsinan glaciation in at least two refugia: (i) the Bering Refuge north of the ice sheets; and (ii) either the Upper Missouri or the southwest Alberta Refuge, south of the ice sheet.     

Isolation of Polymorphic RAPD-SSR Markers from Xinjiang Arctic Grayling (<Emphasis Type="Italic">Thymallus arcticus grubei</Emphasis>) and a Test of Cross-Species Amplification

A total of 18 polymorphic microsatellite loci were isolated and characterized from RAPD products in the Xinjiang Arctic Grayling (Thymallus arcticus grubei). The number of alleles (N_a) per locus varied from 2 to 10. Observed (H_o) and expected (H_e) heterozygosities ranged from 0.64 to 0.92, and from 0.63 to 0.88, respectively. Considerable differences were found among HBH, FH and FY populations in the number of alleles, effective number of alleles, number of genotypes at all of these loci. These new RAPD-SSR markers have provided a helpful tool for genetic analyses and resources conservation of T. arcticus grubei. Five additional fish species, Amur grayling (Thymallus grubii), Taimen (Hucho taimen), Sea perch (Lateolabrax japonicus), Lenok (Brachymystax lenok) and Red seam bream (Pagrosomus major) were assessed for cross-species amplification. Three of the five species showed at least one polymorphic locus. In addition, seven loci were found to be polymorphic in at least one species.     

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.     

Analysis of ribosomal RNA intergenic spacer (IGS) sequences in species and populations of Gyrodactylus (Platyhelminthes: Monogenea) from salmonid fish in Northern Europe

The intergenic spacer (IGS) region of ribosomal RNA genes was amplified and sequenced from a variety of Gyrodactylus specimens collected from wild and farmed Atlantic salmon Salmo salar, rainbow trout Oncorhynchus mykiss, and grayling Thymallus thymallus, from various locations in Northern Europe. Phylogenetic analysis of the sequences confirmed the distinction between G. salaris Malmberg, 1957 and G. thymalli Zitnan, 1960, supporting their validity as separate species. G. salaris adapted to rainbow trout are also distinct from the parasites found on Atlantic salmon, supporting the existence of a rainbow-trout form that was initially identified on the basis of morphological differences. Analysis of the IGS did not provide good resolution of different populations of G. salaris sensu stricto, but was consistent with epidemiological evidence which indicates that introduction of the parasite to Norway was recent and limited. The IGS may be helpful in distinguishing forms of G. salaris that are pathogenic to Atlantic salmon from those that are not.     

The susceptibility of grayling (Thymallus thymallus) to experimental infections with the monogenean Gyrodactylus salaris

The pathogenic monogenean Gyrodactylus salaris infecting Atlantic salmon (Salmo salar) is found to attach and reproduce under laboratory conditions on several species in the subfamily Salmoninae other than the Atlantic salmon. The gyrodactylid species Gyrodactylus thymalli infecting grayling (Thymallus thymallus) in another subfamily, Thymallinae, is previously said to be very similar to G. salaris based on morphometry and genetical analysis which prompted the present laboratory experiments to test the susceptibility and resistance of grayling to G. salaris. All 0+ and 1+ grayling became infected with G. salaris during the experimental infection procedure. However, both innate resistant and susceptible grayling were found. In susceptible individually isolated fish, parasite reproduction lasted for more than 35 days. Parasite reproduction also occurred among grouped grayling as judged from the duration of infection of more than 50 days. However, grayling susceptibility as judged from G. salaris reproduction, was very limited. Hence, the results indicate significant biological differences between the function of Atlantic salmon and grayling as host for G. salaris. The grayling is interpreted as unable to sustain G. salaris in nature which implies that G. thymalli is not conspecific with G. salaris. However, G. salaris dispersal by grayling cannot be excluded.     

Cytochemical characteristics of the labrocyte-like cells in the capsule of the plerocercoids of Triaenophorus nodulosus (Pallas, 1781) and Diphyllobothrium dendriticum (Nitzsch, 1824) cestodes]

Out of six fish species examined (Esox lucius, Paracottus kessleri, Perca fluviatilis, Leuciscus leuciscus baicalensis, Coregonus autumnalis migratorius, Thymallus arcticus baicalensis) labrocyte-like cells were detected in Esox lucius, Leuciscus leuciscus baicalensis, Coregonus autumnalis migratotius. The cells contain large metachromatic granules. Histochemical methods revealed in them sulfated and carboxylic acid mucopolysacharides. The amount of labrocyte-like cells in the capsules around plerocercoids of Triaenophrus nodulosis from Esox lucits liver depends on the age of the capsule and that of the plerocercoid. It increases when the connective tissue papsule is forming and decreases again under aging and degeneration of the capsule containing plerocercoid. The amount of labrocyte-like cells in the cestodes capsules of Raillietina increases sharply when they locate in an accidental organ (liver). This demonstrates the acuteness of organ reactivity to a nonspecific helminth.     

Initial steps of speciation by geographic isolation and host switch in salmonid pathogen Gyrodactylus salaris (Monogenea: Gyrodactylidae)

To test the hypothesis that host-switching can be an important step in the speciation of gyrodactylid monogenean flatworms, we inferred the phylogeny within a cluster of parasites morphologically close to Gyrodactylus salaris Malmberg 1957, collected from Atlantic, Baltic and White Sea salmon (Salmo salar), farmed rainbow trout (Oncorhynchus mykiss), and grayling (Thymallus thymallus) from Northern Europe. The internal transcribed spacer region of the nuclear ribosomal gene was sequenced for taxonomic identification. Parasites on grayling from the White Sea Basin differed from the others by one nucleotide (0.08%), the remainder were identical to the sequence published earlier from Norway (G. salaris on salmon), England (Gyrodactylus thymalli on grayling), and the Czech Republic (unidentified salaris/thymalli on trout). For increased resolution, 813 nucleotides of the mitochondrial COI gene of 88 parasites were sequenced and compared with 76 published sequences using phylogenetic analysis. For all tree building algorithms (NJ, MP), the parasites formed a star-like phylogeny of six definite sister clades, indicating nearly simultaneous radiation. Average K2P distances between clades were 1.8-2.6%, and internal mean distances 0.2-1.1%. The genetic distance to the nearest known relative, Gyrodactylus lavareti Malmberg, was 24%. A variable salmon-specific mitochondrial Clade I was observed both in the Baltic Basin and in pathogenic populations introduced to the Atlantic and White Sea coasts. An invariable Clade II was common in rainbow trout farms in Sweden, Denmark and Finland; the same haplotype was also infecting salmon in a landlocked population in Russian Karelia, and in Oslo fjord and Sognefjord in Norway. Four geographically vicariant sister clades were observed on graylings: Clade III in the Baltic Sea Basin; Clade IV in Karelian rivers draining to the White Sea; Clade V in Norwegian river draining to Swedish lake V?nern; and Clade VI in rivers draining to Oslo fjord. The pattern fitted perfectly with the postglacial history of grayling distribution. Widely sampled clades from salmon and Baltic grayling had basal haplotypes in populations, which were isolated early during the postglacial recolonisation. The divergence between the six clades was clear and linked with their hosts, but not wide enough to support a species status for them. Parasites from the Slovakian type population of G. thymalli were not available, so this result does not mean that G. salaris and G. thymalli are synonyms. It is suggested that the plesiomorphic host of the parasite cluster was grayling, and the switch to salmon occurred at least once when the continental ice isolated Baltic salmon in an eastern freshwater refugium, 130,000 years ago. At the same time, parasites on grayling were split geographically and isolated into several allopatric refugia. The divergence among the parasite clades allowed a tentative calibration of the evolutionary rate, leading to an estimate of the divergence of 13.7-20.3% per million years for COI coding mtDNA. The results supported the hypothesis that parallel to the allopatric mode, host switch and instant isolation by host specificity can be operated as a speciation mechanism.     

Genetic lineages and postglacial colonization of grayling (Thymallus thymallus, Salmonidae) in Europe, as revealed by mitochondrial DNA analyses

In stark contrast to other species within the Salmonidae family, phylogeographic information on European grayling, Thymallus thymallus, is virtually nonexistent. In this paper, we utilized mitochondrial DNA polymerase chain reaction-restriction fragment length polymorphism (mtDNA PCR-RFLP) and sequence variation to infer the postglacial dispersal routes of T. thymallus into and within northern Europe, and to locate geographically, potential evolutionarily distinct populations. Mitochondrial analyses revealed a total of 27 T. thymallus haplotypes which clustered into three distinct lineages. Average pairwise interlineage divergence was four and nine times higher than average intralineage divergence for RFLP and sequence data, respectively. Two European grayling individuals from the easternmost sample in Russia exhibited haplotypes more genetically diverged from any T. thymallus haplotype than T. arcticus haplotype, and suggested that hybridization/introgression zone of these two sister species may extend much further west than previously thought. Geographic division of the lineages was generally very clear with northern Europe comprising of two genetically differentiated areas: (i) Finland, Estonia and north-western Russia; and (ii) central Germany, Poland and western Fennoscandia. Average interpopulation divergence in North European T. thymallus was 10 times higher than that observed in a recent mtDNA study of North American T. arcticus. We conclude that (i) North European T. thymallus populations have survived dramatic Pleistocene temperature oscillations and originate from ancient eastern and central European refugia; (ii) genetic divergence of population groups within northern Europe is substantial and geographically distinct; and (iii) the remainder of Europe harbours additional differentiated assemblages that likely descend from a Danubian refugium. These findings should provide useful information for developing appropriate conservation strategies for European grayling and exemplify a case with a clear need for multinational co-operation for managing and conserving biodiversity.